MXPA05014102A

MXPA05014102A - Engineering single-gene-controlled staygreen potential into plants

Info

Publication number: MXPA05014102A
Application number: MXPA/A/2005/014102A
Authority: MX
Inventors: R Gallie Daniel; Meeley Robert; Young Todd
Original assignee: R Gallie Daniel; Meeley Robert; Pioneer Hibred International Inc; The Regents Of University Of California; Young Todd
Priority date: 2003-06-23
Filing date: 2005-12-20
Publication date: 2006-10-17

Abstract

The enzymes of the ACC synthase family are used in producing ethylene. Nucleotide and polypeptide sequences of ACC synthases are provided along with knockout plant cells having inhibition in expression and/or activity in an ACC synthase and knockout plants displaying a staygreen phenotype, a male sterility phenotype, or an inhibition in ethylene production. Methods for modulating staygreen potential in plants, methods for modulating sterility in plants, and methods for inhibiting ethylene production in plants are also provided.

Description

ENGINEERING POTENTIAL OF GREEN PERMANENCE CONTROLLED BY A SINGLE GENE IN PLANTS FIELD OF THE INVENTION This invention relates to the modulation of the green permanence potential in plants, the inhibition of ethylene production in plants, and the modulation of sterility in plants. The invention also provides cells from inactive plants, for example, where the cells of inactive plants are disrupted in the expression and / or activity of ACC synthase, or inactive plants, for example, that exhibit a green permanence phenotype or a sterility phenotype. masculine Also included are nucleic acid sequences and amino acid sequences encoding various ACC synthases. BACKGROUND OF THE INVENTION The green permanence is a term used to describe a plant phenotype, for example, whereby the leaf's senescence (more easily distinguished by the leaf aridity associated with the chlorophyll degradation) is retarded. compared to a standard reference. See, Thomas H and Howarth CJ (2000) "Yive ways to stay green" Journal of Experimental Botany 51: 329-337 In sorghum, several genotypes of green permanence have been identified that exhibit a delay in leaf senescence during the filling of the grain and ripening. See, Duncan RR, et al., (1981) "Descriptive compar ± son of senescent and non-senescent sorghum g eno type s". Agronomy Journal 73: 849-853. In addition, under conditions of limited water availability, which normally hastens the senescence of the leaf (see, for example, Rosenow DT and 'Clark LE (1981) Drought tolerance in sorghum .. In: Loden HD, Wilkinson D, eds. of the 36th annual corn and sorghum industry research conference, 18-31), these genotypes retain more area of the green leaf and continue to fill the grain normally (see, for example, McBee GG, Waskom RM, Miller FR, Creelman RA (1983 ) Effect of senescence and non-senescence on carbohydrates in sorghum during late kernel maturity states Crop Science 23: 372-377, Rosenow DT, Quisenberry JE, Wendt CW, Clark LE (1983) Drought-tolerant sorghum and cotton germplasm. Management 7: 207-222; and, Borrell AK, Douglas ACL (1996) Maintaining green leaf area in grain sorghum increases yield in a water-limited environment In: Foale MA, Henzell RG, Kneipp JF, eds Proceedings of the third Australian sorghum conference Melbourne: Australian Inst Itute of Agricultural Science, Occasional Publication No. 93). The green permanence phenotype has also been used as a selection criterion for the development of improved maize varieties, particularly with respect to the development of drought tolerance. See, for example, Russell WA (1991) Genetic improvement of maize yields. Advances in Agronomy 46: 245-298; and Bruce et al., (2002), "Molecular and physiological approaches to maize improvement for drought tolerance" Journal of Experimental Botany, 53 (366): 13-25. Five fundamentally different types of green permanence have been described, which are types A, B, C, D and E (see, for example, Thomas H, Sraart CM (1993) Crops that stay green, Annals of Applied, Biology 123: 193 -219; and Thomas and Howarth, supra). In the green type A stay, the initiation of the old age program is delayed, but then proceeds at a normal rate. In the green type B stay, while the initiation of the senescence program is unchanged, the progression is comparatively slower. In green Type C permanence, chlorophyll is retained although senescence (as determined by measurements of physiological function such as photosynthetic capacity) proceeds at a normal rate. The green permanence of type D is more artificial in that the extermination of the leaf (that is, by freezing, boiling or drying) prevents the initiation of the senescence program, thus halting the degradation of chlorophyll. In green type E, the initial levels of chlorophyll are higher, while the initiation and progression of the leaf's senescence are unchanged, thus giving the illusion of a higher proportion. of relatively slow progression. Type A and B are functional green stays, since the photosynthetic capacity is maintained together with the chlorophyll content, and these are the types associated with increased yield and tolerance to drought in sorghum. Despite the potential importance of this attribute, particularly the benefits associated with increased yield and tolerance to drought, very little progress has been made in understanding the biochemical, physiological or molecular basis for genetically determined green permanence ( Thomas and Howarth, supra). This invention solves these and other problems. The invention relates to the identification of the ACC synthase genes associated with the phenotype of green permanence potential in plants and the modulation of the green permanency potential and / or the production of ethylene. The polypeptides encoded by these genes, methods to modulate the green staying potential in plants, methods to inhibit the production of ethylene in plants, methods to modulate sterility in plants and cells of inactive plants and plants, as well as other characteristics, will reach Be evident in the review of the following materials. BRIEF DESCRIPTION OF THE INVENTION This invention provides methods and compositions for modulating the green permanence potential and the sterility in plants and modulate (for example, inhibit) ethylene synthesis and / or plant production. This invention also relates to nucleic acid sequences of ACC synthase in plants, exemplified by, for example, SEQ ID NO: 1 to SEQ ID NO: 6 and SEQ ID NO: 10, and a set of polypeptide sequences, for example , SEQ ID NO: 7 to SEQ ID NO: 9 and SEQ ID NO: 11, which can modulate these activities. In a first aspect, the invention provides an isolated or recombinant inactive plant cell comprising at least one interruption in at least one endogenous ACC synthase gene (e.g., a nucleic acid sequence, or complement thereof, comprising , for example, at least about 70%, at least about 75%, at least about 80%, - at least about 85%, at least about 90%, at least about 95%, at least about 99%, about 99.5% or more of sequence identity to SEQ ID NO: l (gACS2), SEQ ID NO: 2 (gACS6) or SEQ ID NO: 3 (gACS7)). The disruption inhibits the expression or activity of at least one ACC synthase protein compared to a corresponding control plant cell that lacks the interruption. In one embodiment, the at least one endogenous ACC synthase gene comprises two or more endogenous ACC synthase genes (e.g., any two or more of ACS2, ACS6 and ACS7, for example, ACS2 and ACS6). Similarly, in another embodiment, the at least one endogenous ACC synthase gene comprises three or more endogenous ACC synthase genes. In certain embodiments, the disruption results in the production of reduced ethylene by the inactive plant cell as compared to the control plant cell. In one embodiment, the at least one interruption in the inactive plant cell is produced by introducing at least one polynucleotide sequence comprising a nucleic acid sequence of ACC synthase, or subsequence thereof, into a plant cell, such that at least one polynucleotide sequence is linked to a promoter in a sense or antisense orientation, and wherein the at least one polynucleotide sequence comprises, for example, at least about 70%, at least about 75 %, at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 99%, about 99.5% or more sequence identity to SEQ ID NO : l (gACS2), SEQ ID NO: 2 (gACS6), SEQ ID NO: 3 (gACS7), SEQ ID NO: 4 (cACS2), SEQ ID NO: 5 (cACSd), SEQ ID NO: 6 (cACS7) or SEQ ID NO: 10 (CCRA178R) or a subsequence thereof, or a complement thereto. In another modality, the interruption is introduced in the plant cell by introducing at least one polynucleotide sequence comprising one or more subsequences of an ACC synthase nucleic acid sequence configured for silencing or RNA interference. In another embodiment, the interruption comprises the insertion of one or more transposons, wherein the one or more transposons are in at least one endogenous ACC synthase gene. In yet another embodiment, the interruption comprises one or more point mutations in at least one endogenous ACC synthase gene. The interruption may be a homozygous interruption in at least one ACC synthase gene. Alternatively, the interruption is a heterozygous interruption in at least one ACC synthase gene. In certain embodiments, when more than one ACC synthase gene is involved, there is more than one interruption, which may include homozygous interruptions, heterozygous interruptions or a combination of homozygous interruptions and heterozygous interruptions. In certain embodiments, a plant cell of the invention is of a dicot or monotone. In one aspect, the plant cell is in a hybrid plant comprising a green permanence potential phenotype. In another aspect, the plant cell is in a plant comprising a sterility phenotype, for example, a male sterility phenotype. The regenerated plants of The plant cells of the invention are also a feature of the invention. The invention also provides inactive plants comprising a phenotype of green residence potential. For example, the invention provides an inactive plant comprising a phenotype of green permanence potential, where the phenotype of green permanency potential results from an interruption in at least one endogenous ACC synthase gene. In one embodiment, the interruption includes one or more transposons, and inhibits the expression or activity of at least one ACC synthase protein compared to a corresponding control plant. In another embodiment, the disruption includes one or more point mutations in the endogenous ACC synthase gene and inhibits the expression or activity of at least one ACC synthase protein compared to a corresponding control plant. In certain embodiments, the at least one endogenous ACC synthase gene comprises a nucleic acid sequence comprising, for example, at least about 70%, at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 99? S, about 99.5% or more, of sequence identity to SEQ ID NO: l (gACS2), SEQ ID NO: 2 (gACS6) or SEQ ID NO: 3 (gACS7), or a complement thereof. In certain modalities, the inactive plant is a hybrid plant. Essentially all the characteristics mentioned in the above apply to this modality also, as it is relevant. In another embodiment, an inactive plant includes a transgenic plant comprising a green permanence potential phenotype. For example, a transgenic plant of the invention includes a phenotype of green permanency potential that results from at least one introduced transgene that inhibits ethylene synthesis. The introduced transgene includes a nucleic acid sequence encoding at least one ACC synthase or subsequence thereof, the nucleic acid sequence comprising, for example, at least about 70%, at least about 75%, so less about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 99%, about 99.5% or more of sequence identity to SEQ ID NO: 1 ( gACS2), SEQ ID NO: 2 '(gACSd), SEQ ID NO: 3 (gACS7), SEQ ID N0: 4 (cACS2), SEQ ID NO: 5 (cACS6), SEQ ID NO: 6 (CACS7) or SEQ ID NO: 10 (CCRA178R), or a subsequence thereof, as a complement thereto, and is in a configuration that modifies an expression or activity level of the at least one ACC synthase (e.g. sense, antisense, silencing or RNA interference). Essentially all the features mentioned above apply to this modality as well, as relevant. A transgenic plant of the invention may also include a green permanency potential phenotype that results from at least one introduced transgene that inhibits ethylene synthesis, wherein at least one introduced transgene comprises a nucleic acid sequence encoding one (s). ) subsequence (s) of at least one ACC synthase, with at least one ACC synthase comprising, for example, at least about 70%, at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 99%, about 99.5% or more of sequence identity to SEQ ID NOY (pACS2), SEQ ID NO: 8 (pACSd) ), SEQ ID NO: 9 (pACS7) or SEQ ID NO.11 (pCCRA178R), or a conservative variation thereof. The nucleic acid sequence is typically in a configuration of RNA silencing or interference (or for example, a sense or antisense configuration) and modifies a level of expression or activity of the at least one ACC synthase.

Essentially all the characteristics mentioned in the above apply to this modality also, as it is relevant.

The green permanency potential of a plant of the invention includes, but is not limited to, for example, (a) a reduction in the production of at least one specific mRNA of ACC synthase; (b) a reduction in the production of an ACC synthase; (c) a reduction in ethylene production; (d) a delay in the senescence of the leaf; (e) an increase in drought resistance; (f) an increased time in maintaining the photosynthetic activity; (g) increased perspiration; (h) an increased stomatal conductance; (i) an increased assimilation of CO; (j) an increased assimilation maintenance of C02; or (k) any combination of (a) - (j); compared to a corresponding control plant and the like. One aspect of the invention provides inactive or transgenic plants that include sterility phenotypes, for example, a male or female sterility phenotype. Thus, one class of modalities provides an inactive plant comprising a male sterility phenotype (e.g., reduced pollen spread) resulting from at least one interruption in at least one endogenous ACC synthase gene. The disruption inhibits the expression or activity of at least one ACC synthase protein compared to a corresponding control plant.

In one embodiment, the at least one interruption results in the production of reduced ethylene by the inactive plant as compared to the control plant. In one embodiment, the at least one interruption includes one or more transposons, wherein the one or more transposons are in at least one endogenous ACC synthase gene. In another embodiment, the at least one interruption comprises one or more point mutations, wherein the one or more point mutations are in at least one endogenous ACC synthase gene. In yet another embodiment, the at least one interruption is introduced into the inactive plant by introducing at least one polynucleotide sequence comprising one or more subsequences of an ACC synthase nucleic acid sequence configured for RNA silencing or interference. In certain embodiments, the at least one endogenous ACC synthase gene comprises a nucleic acid sequence comprising, for example, at least about 70%, at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 99%, about 99.5% or more, of sequence identity to SEQ ID? 0: 1 (gACS2), SEQ ID? O: 2 (gACS6) or SEQ ID? O: 3 (gACS7), or a complement thereof. Essentially all the characteristics mentioned in the above apply to this modality also, as it is relevant. Another kind of modalities provides a plant transgenic inactive comprising a male sterility phenotype resulting from at least one introduced transgene that inhibits ethylene synthesis. The at least one introduced transgene comprises a nucleic acid sequence encoding at least one ACC synthase, the nucleic acid sequence comprising at least about 85% sequence identity at SEQ ID NO: 1 (gACS2), SEQ ID NO: 2 (gACS6), SEQ ID NO: 3 (gACS7), SEQ ID NO: (CACS2), SEQ ID NO: 5 (cACSβ), SEQ ID NO: 6 (cACS7) or SEQ ID NO: 10 (CCRA178R ), or a subsequence thereof, or a complement thereto, and is in a configuration that modifies a level of expression or activity of the at least one ACC synthase (e.g., an antisense, sense or silencing or RNA interference). In certain embodiments, the transgene includes a tissue-specific promoter or an inducible promoter. Essentially all the characteristics mentioned in the above apply to this modality also, as it is relevant. Polynucleotides are also a feature of the invention. In certain embodiments, a recombinant isolated polynucleotide comprises a member selected from the group consisting of: (a) a polynucleotide, or a complement thereof, comprising, for example, at least about 70%, at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 99%, about 99.5% or more of sequence identity to SEQ ID NO: l (gACS2), SEQ ID NO: 2 (gACS6), SEQ ID NOY (gACS7), SEQ ID NO: 4 (cACS2), SEQ ID NO: 5 (cACS6), SEQ ID NO: 6 (cACS7) or SEQ ID NO: 10 (CCRA178R), or a subsequence thereof, or a conservative variation thereof; (b) a polynucleotide, or a complement thereof, that encodes a polypeptide sequence of SEQ ID NOY (pACS2), SEQ ID NO: 8 (pACS6), SEQ ID NO: 9 (pACS7) or SEQ ID NO: 11 (pCCRA178R), or a subsequence thereof, or a conservative variation thereof; (c) a polynucleotide, or a complement thereof, that hybridizes under severe conditions over substantially the entire length of a polynucleotide subsequence comprising at least 100 contiguous nucleotides of SEQ ID NO: 1 (gACS2), SEQ ID NO : 2 (gACSß), SEQ ID NO: 3 (gACS7), SEQ ID NO: 4 (cACS2), SEQ ID NO: 5 (cACSβ), SEQ ID NO: 6 (cACS7) or SEQ ID NO: 10 (CCRA178R), or hybrid a sequence of polynucleotide of (a) or (b); and, (d) a polynucleotide that is at least about 85 ° identical to a polynucleotide sequence of (a), (b) or (c). In certain embodiments, the polynucleotide inhibits the production of ethylene when expressed in a plant. The polynucleotides of the invention can understand or be contained within an expression cassette or a vector (eg, a viral vector). The expression vector or cassette may comprise a promoter (eg, a constitutive, tissue-specific or inducible promoter) or operably linked to the polynucleotide. A polynucleotide of the invention can be linked to the promoter in an amphisense orientation or a sense orientation, can be configured for silencing or RNA interference, or the like, The invention also provides methods to inhibit the production of ethylene in a plant (and plants produced by such methods). For example, a method for inhibiting ethylene production comprises inactivating one or more ACC synthase genes in the plant, wherein the one or more ACC synthase genes encode one or more ACC synthases, wherein at least one of the one or more ACC synthases comprises, for example, at least about 70%, at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 99%, approximately the identity of 99.5% or more identity to the SEQ ID NOY (pACS2), SEQ ID NO: 8 (pACSd), SEQ ID N0: 9 (pAC7) or SEQ ID NO: 11 (pCCRA178R ). In one embodiment, the inactivation step comprises introducing one or more mutations into a sequence of ACC synthase gene, wherein the one or more mutations in the ACC synthase gene sequence comprises one or more transposons, to thereby inactivate the one or more ACC synthase genes compared to a corresponding control plant. In another embodiment, the inactivation step comprises introducing one or more mutations into an ACC synthase gene sequence, wherein the one or more mutations in the ACC synthase gene sequence comprise one or more point mutations, in order to inactivate the one or more ACC synthase genes compared to a corresponding control plant. The one or more mutations may comprise, for example, a homozygous disruption in the one or more ACC synthase genes, a heterozygous interruption in the one or more ACC synthase genes, or a combination of both omocictive interruptions and heterozygous interruptions if more than one gene ACC synthase is interrupted. In certain modalities, the one or more mutations are introduced by a sexual cross. In certain embodiments, at least one of the one or more ACC synthase genes is, for example, at least about 70%, at least about 75%, at least about 80%, at least about 85%, so less about 90%, at least about 95%, at least about 99%, about 99.5% or more, identical to SEQ ID NO: 1 (gACS2), SEQ ID NOY (gACSd) or SEQ ID NO: 3 ( gAC7), or a complement of them). In another embodiment, the inactivating step comprises: (a) introducing into the plant at least one polynucleotide sequence, wherein at least one polynucleotide sequence comprises a nucleic acid encoding one or more ACC synthases, or a subsequence of the same, and a promoter, the promoter that 'works in the plants to produce an RNA sequence; and, (b) expressing the at least one polynucleotide sequence, thereby inactivating the one or more ACC synthase genes compared to a corresponding control plant (e.g., its non-transgenic origin or a non-transgenic plant thereof). species). For example, the at least one polynucleotide sequence can be introduced by techniques including, but not limited to, electroporation, micro-projectile bombardment, Agrobacterium-mediated transfer, and the like. In certain aspects of the invention, the polynucleotide is linked to the promoter in a sense orientation or an antisense orientation, or is configured for RNA silencing or interference. Essentially all the characteristics mentioned in the above apply to this modality also, as it is relevant. Methods for modulating the potential for green permanence in plants are also a feature of the invention (such as plants produced by such plants). methods). For example, a method for modulating the green residence potential comprises: a) selecting at least one ACC synthase gene (eg, encoding an ACC synthase, e.g., SEQ ID NOY (pACS2), SEQ ID N0: 8 ( pACSd), SEQ ID NO: 9 (pAC7) or SEQ ID NO: 11 (pCCRA178R)) to mutate, thereby providing at least one desired ACC synthase gene; b) introducing a mutant form (e.g., an antisense or sense configuration of at least one ACC synthase gene or subsequence thereof, an RNA silencing configuration of at least one ACC synthase gene or subsequence thereof, a heterozygous mutation) in at least one ACC synthase gene, a mutation homozygous in the at least one ACC synthase gene or a combination of homozygous mutation and heterozygous mutation if more than one ACC synthase gene, and the like) of at least one gene is selected ACC desired synthase in the plant; and, c) express the mutant form, in order to modulate the potential for green permanence in the plant. In one embodiment, the selection of at least one ACC synthase gene comprises determining a degree (eg, weak, moderate or strong) of desired green residence potential. In certain embodiments, the mutant gene is introduced by Agrobacterium-mediated transfer, electroporation, micro-projectile bombardment, a sexual cross, or the like. Essentially all the features mentioned in the above apply to this modality also, as relevant. The detection of expression products is done either qualitatively (by detecting the presence or absence of one or more products of interest) or quantitatively (by monitoring the level of expression of one or more products of interest). In one embodiment, the expression product is a product of RNA expression. Aspects of the invention optionally include monitoring a level of expression of a nucleic acid, polypeptide or chemical substance (eg, ACC, ethylene, etc.) as mentioned herein for the detection of ACC synthase, ethylene production, potential green permanence, etc. in a plant or in a population of plants. The compositions and methods of the invention may include a variety of plants, for example, a plant of the family Poaceae (Gramineae). Examples of members of the Poaceae family include, but are not limited to, Acampt ociates, Achnatherum, Achnella, Acroceras, Aegilops, Aegopgon, Agroelymus, Agrohordeum, Agropogon, Agropyron, Agrosi tanion, Agrostis, Aira, Allolepis, Alloteropsis, Alopecurus, Amblyopyrum , Ammophila, Anapelodesms, Amphibromus, Arnphicarpum, Anzphilophis, Anastrophus, Anatherum, Andropogron, Anemathele, Aneurolepidium, Anisantha, Anthaenantia, Aizthephora, Ayzthochloa, Anthoxanthunz, Apera, Apluda, Archtagrostis, Arctophila, Argillochloa, Aristida, Arrhenatherum, Arthraxon, Arthrostylidiufn, Arundiyzaria, Arundinella, Arundo, Aspris, Atheropogon, Oats, Avenella, Avenochloa, Avenula, Axonopus, Bambusa, Beckmannia, Blepharidachne, Blepharoneuron, Bothriochloa, Bouteloua, Brachiaria, Brachyelytrum, Brachypodium, Briza, Brizopyrum, Bromelica, Bromopsis, Bromus, Buchloe, Bulbilis, Calamagrostis, Calamovilfa, Campulosus, Capriola, Catabrosa, Catapodum, Cathestecum, Cenchropsis, Cenchrus, Centotheca, Ceratochloa, Chaetochloa, Chasmanthium, Chimonobambusa, Chionochloa, Chloris, Chondrosum, Chrysopon, Chusquea, cinna, Cladoraphis, Coelorachis, Coix, Coleanthus, Colpodium, Coridochloa, Conaucopiae, Cortaderia, Corynephorus, Cottea, Cri tesion, Crypsis, Ctenium, Cutandia, Cylindropyrum, Cymbopogon, Cynodon, Cynosurus, Cytrococcum, Dactylis, Dactyloctenium , Danthonia, Dasyochloa, Dasyprum, Davyella, Dendr ocal am s, Deschampsia, Desmazeria, Deyeuxia, Diarina, diarrhena, Dichantlaelium, Dichantlaium, Dichelachne, Diectomus, Digitaria, Dimeria, Dimorpostachys, Dinebra, Díplachne, Dissantlieliutii, Dissochondrus, Distichlis, Drepanostacllyuni, Dupoa, Dupontia, Echinochloa, Ectosperm a, Elirharta, Eleusine, Elyhordeum, Elyleymus, elymordeum, Elymus, Elyonurus, Elysitanion, Elytesion, Elytrigia, Enneapogon, Enteropogon, Epicampes, Eragrostis, Eremochloa, Ereiiiopoa, Eremopyrum, Erianthus, Ericoma, Erichloa, Eriochrysis, Erioneuron, Euchlaena, Euclasta, Eulalia, Eulaliopsis, Eustachys, Fargesia, Festuca, Festuloliu, Fingerhuthia, Fluminia, Garnotia, Gastridium, Gaudinia, Gigantochloa, Glyceria, Graphephorum, gymnopogon, Gynerium, Hackelochloa, Hainardia, Hakonechloa, Haynaldia, Heleochloa, Helictotrichon, Hemarthria, Hesperochloa, Hesperostipa, Heteropogon, Hibanobanzbusa, Hierochloe, Hilaria, Holcus, Homalocenchrus, Hordeum, Hydrochloa, Hymenachne, Hyparrhenia, Hypogynium, Hystrix, Ichnanthus, Imperata, Indocalamus, Isachne, Ischaemum, Ixophorus, Koelerla, Korycarpus, Lagurus, Lanaarckia, Lasiacis, Leersia, Leptochloa, Leptochloopsis, Leptocoryphium, Leptoloma, Leptogon, Lepturus, Lerchenfeldia, Leucopoa, Leymostachys, Leymus, Limnodea, Li thachne, Lolium, Lophochlaena, Lophochloa, Lophopyruta, Ludolfia, Luziola, Lycurus, Lygeum, Maltea, Manisuris, Megastachya, Melica, Melinis, Mibora, Microchloa, Microlaena, Microstegium, Milium, Miscanthus , Mnesithea, Molinia, Monanthochloe, Monerma, Monroa, Muhlenbergia, Nardus, Nassella, Nazia, Neeragrostis, Neoschischkinia, Neostapfia, Neyraudia, Nothoholcus, Olyra, Opizia, Oplismenus, Orcuttia, Oryza, Oryzopsis, Otatea, Oxytenanthera, Panicularia, Panicum, Pappophorum, Parapholis, Pascopyrum, Paspalidium, Paspalum, Pennisetum, Phalaris, Phalaroides, Phanopyrum, Pharus, Phippsia, Phleum, Pholi urus, Phragmi tes, Phyllostaclzys, Piptatherum, Piptochaetium, Pleioblastus, Pleopogon, Pleuraphis, Pleuropogon, Poa, Podagrostis, Polypogon, Polytrias, Psathyrostaclzys, Pseudelymus, Pseudoroegneria, Pseudosasa, Ptilagrostis, Puccinellia, Pucciphippsia, Redfieldia, Reinzaria, Reimarochloa, Rhaphis, Rhombolytrum, Rhynchelytrum, Roegneria, Rostraria, Rottboellia, Rytilix, Saccharum, Sacciolepis, Sasa, Sasaella, Sasamorpha, Savastana, Schedonnardus, Schismus, Schizachne, Schizachyrium, Schizostachyum, Sclerochloa, Scleropoa, Scleropogon, Scolochloa, Scribneria, Sécale, Semiarundinaria, Sesleria, Setaria, Shibataea, Sieglingia, Sinundundinaria, Sinobambusa, Sinocalamus, Sitanion, Sorghastrum, Sorghum, Spartina, Sphenopholis, Spodiopogon, Sporobolus, Stapfia, Steinchisma, Stenotaphrum, Stipa, Stipagrostis, Stiporyzopsis, Swallenia, Syntherisma, Taeniatherum, Terrellia, Terrelymus, Thamnocalamus, Themeda, Thinopyrum, Thuarea, Thysalzolaella, Torresia, Torreyochloa, Trachynia, Trachypogofa, Tragus, Trichachne, Trichloris, Triclzolaena, Trichoneura, Tridens, Triodia, Triplasis, Tripogon, Tripsacum, Trisetobromus, Trisetum, Triticosecale, Triticum, Tuctoria, Unióla, Urachne, Uralepis, Urochloa, Vahlodea, Valota, Vaseyochloa, Ventenata, Vetiveria, Vilfa, Vulpia, Willkommia, Yushania, Zea, Zizania, Zizaniopsis and Zoysía. In one modality, the plant is Zea mays, wheat, rice, sorghum, barley, oats, turf grass, rye, soy, tomato, potato, pepper, broccoli, cabbage, a line of commercial corn, or the like. The equipment that incorporates one or more of the nucleic acids or polypeptides mentioned in the above is also a feature of the invention. Such equipment may include any of the components mentioned in the foregoing and further include, for example, instructions for the use of the components in any of the methods mentioned herein, packaging materials, containers for containing the components and / or the like . For example, a device for modulating green residence potential in a plant includes a container that contains at least one polynucleotide sequence comprising a nucleic acid sequence, wherein the nucleic acid sequence is, for example, at least about 70%, at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 99%, about 99.5% or more , identical to SEQ ID NO: l (gACS2), SEQ ID NO: 2 (gACSd), SEQ ID NO: 3 (gACS7), SEQ ID NO: 4 (CACS2), SEQ ID NOY (cACSd), SEQ ID NO: 6 (cAC7) or SEQ ID NO: 10 (CCRA178R), or a subsequence thereof, or a complement thereto. In an additional modality, the equipment includes instructional materials for the use of minus a polynucleotide sequence to control the green residence potential in a plant. Essentially all the characteristics mentioned in the above apply to this modality also, as it is relevant. Co or another example, a device for modulating sterility, for example, male sterility, in a plant includes a container containing at least one polynucleotide sequence comprising a nucleic acid sequence, wherein the nucleic acid sequence is , for example, at least about 70%, at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 99%, approximately 99.5% or more, identical to SEQ ID NO: l (gACS2), SEQ ID NOY (gACSd), SEQ ID NO: 3 (gACS7), SEQ ID NOY (cACS2), SEQ ID NO: 5 (cACSd), SEQ ID NO: 6 (cAC7) or SEQ ID NO: 10 (CCRA178R), or a subsequence thereof, or a complement thereto. The kit optionally includes instructional materials for the use of the at least one polynucleotide sequence to control sterility, eg, male sterility, in a plant. Essentially all the characteristics mentioned in the above apply to this modality also, as it is relevant. DEFINITIONS Before describing the invention in detail, it is to be understood that this invention is not limited to particular biological devices or systems, which may of course vary. It is also to be understood that the terminology used herein is for the purpose of describing particular modalities only, and is not intended to be limiting. As used in this specification and the appended claims, the singular forms "a", "an" and "the" include plural references unless the context clearly dictates otherwise. Thus, for example, the reference to "one cell" includes a combination of two or more cells, and the like. Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which the invention pertains. In the description and the claim of the present invention, the following terminology will be used according to the definitions set forth below. The term "plant" refers generically to any of: whole plants, parts of plants or organs (for example, leaves, stems, roots, etc.), vegetative organs / shoots structures for example leaves, stems and tubers), roots, flowers and floral organs / structures for example bracts, sepals, petals, stamens, carpels, anthers and ovules), seeds (including embryo, endosperm and envelope of the seed), fruit (the mature ovary), plant tissue (for example vascular tissue, terrestrial tissue and the like, tissue culture callus and plant cells ( for example, protective cells, ovules, trichomes and the like and progeny thereof The plant cell, as used herein, further includes, without limitation, cells obtained from or found in: seeds, cultures, suspension cultures, embryos , meristematic regions, callus tissue, leaves, roots, shoots, gametophytes, sporophytes, pollen and microspores Plant cells can also be understood to include modified cells, such as protoplasts, obtained from the tissues mentioned in the above. "Dicotyledonous" refers to a dicotyledonous plant, and dicotyledonous plants belong to a large subclass of Angiosperms that have two seed leaves (cotyledon). The term "monocotyledon" refers to a monocotyledonous plant, which in the development of the plant has only one cotyledon. The term "inactive plant cell" refers to a plant cell that has an interruption in at least one ACC synthase gene in the cell, where the interruption results in reduced expression or activity of the ACC synthase encoded by that gene compared to a control cell. The wonder inactivation may be the result of, for example, antisense constructs, sense constructs, RNA silencing constructs, RNA interference, genomic interruptions (eg, transposons, entanglement, homologous recombination, etc.) and the like. The term "inactive plant" refers to a plant that has an interruption in at least one of its ACC synthase genes in at least one cell. The term "transgenic" refers to a plant that has incorporated nucleic acid sequences, including but not limited to genes, polynucleotides, DNA, and RNA, etc., that has been introduced into a plant compared to a non-introduced plant. The term "endogenous" is related to any gene or nucleic acid sequence that is already present in a cell. A "transposable element" (TE) or "transposable genetic element" is a DNA sequence that can be moved from one location to another in a cell. The movement of a transposable element can occur from episome to episome, from episome to chromosome, from chromosome to chromosome, or from chromosome to episome. The transposable elements are characterized by the presence of inverted repetition sequences in their terminals. Mobilization is mediated enzymatically by a "transposase". Structurally, a transposable element is classified by category as a "transposon" ("TN") or an "insertion sequence element" (IS element) based on the presence or absence, respectively, of genetic sequences in addition to those necessary for the mobilization of the element. A mini-transposon or mini-IS element typically lacks sequences that encode a transposase. The term "nucleic acid" or "polynucleotide" is generally used in its meaning recognized in the art to refer to a ribose nucleic acid (RNA) or deoxyribose nucleic acid polymer (DNA), or analog thereof, for example, a polymer of nucleotides comprising modifications of the nucleotides, a peptide nucleic acid, or the like. In certain applications, the nucleic acid may be a polymer that includes multiple types of monomers, for example, both of RNA and DNA subunits. A nucleic acid can be, for example, a chromosome or chromosomal segment, a vector (e.g., an expression vector), an expression cassette, a naked DNA polymer or RNA, the product of a polymerase chain reaction ( PCR), an oligonucleotide, a probe, etc. A nucleic acid may be, for example, single-stranded and / or double-stranded. Unless otherwise indicated, a particular nucleic acid sequence of this invention optionally comprises or encodes complementary sequences, in addition to any explicitly indicated sequence. The term "polynucleotide sequence" or "nucleotide sequence" refers to a contiguous sequence of nucleotides in an individual nucleic acid or a representation, eg, a string of characters, thereof. That is, a "polynucleotide sequence" is a polymer of nucleotides (an oligonucleotide, a DNA, a nucleic acid, etc.) or a character string that represents a nucleotide polymer, depending on the context. From any specified polynucleotide sequence, either the given nucleic acid or the complementary polynucleotide sequence (e.g., the complementary nucleic acid) can be determined. The term "subsequence" or "fragment" is any portion of a complete sequence. A "phenotype" is the display of an attribute in an individual plant that results from the interaction of gene expression and the environment. An "expression cassette" is a construction of nucleic acid, eg, vector, such as a plasmid, a viral vector, etc., capable of producing transcripts and, potentially, polypeptides encoded by a polynucleotide sequence. An expression vector is capable of producing transcripts in an exogenous cell, for example, a bacterial cell, or a plant cell, in vivo or in vi tro, for example, a protoplast of cultivated plant. The expression of a product can be either constitutive or inducible depending, for example, on the selected promoter. The antisense, sense or interference or RNA silencing configurations that may or may not be translated are expressly included by this definition. In the context of an expression vector, a promoter is said to be "operably linked" to a polynucleotide sequence if it is capable of regulating the expression of the associated polynucleotide sequence. The term also applies constructs of alternative exogenous genes, such as expressed or integrated transgenes. Similarly, the term operably linked equally applies to alternative or additional transcriptional regulatory sequences such as enhancers, associated with a polynucleotide sequence. A polynucleotide sequence is said to "encode" a sense or antisense RNA molecule, or the silencing or interfering molecule of RNA or a polypeptide, if the polynucleotide sequence can be transcribed (in spliced or non-spliced form) and / or translated into the RNA or polypeptide, or a subsequence thereof. "Gene expression" or "expression of an acid "nucleic" means the transcription of DNA into the RNA (optionally including modification of the RNA, eg, splicing), translation of RNA into a polypeptide (possibly including subsequent modification of the polypeptide, eg, post-translational modification) or both transcription and translation, as indicated by context The term "gene" is widely used to refer to any nucleic acid associated with a biological function Genes typically include coding sequences and / or regulatory sequences required for the expression of such sequences The term "gene" applies to a specific genomic sequence, as well as a cDNA or an mRNA encoded by those genomic sequences.The genes also include unexpressed nucleic acid segments that, for example, form recognition sequences for other proteins The regulatory sequences not expressed include promoters and breeders, which regulatory proteins such as transcription factors link, resulting in the transcription of adjacent or neighboring sequences. A "polypeptide" is a polymer that comprises two or more amino acid residues (e.g., a peptide or a protein). The polymer may additionally comprise non-amino acid elements such as brands, quenchers, blocking groups or the like and may optionally comprise modifications such as glycosylation or the like. The amino acid residues of the polypeptide may be natural or unnatural and may be unsubstituted, unmodified, substituted or modified. The term "recombinant" indicates that the material (for example, a cell, a nucleic acid or a protein) has been altered artificially or synthetically (not naturally) by human intervention. The alteration can be made on the material inside, or removed from, its environment or natural state. For example, a "recombinant nucleic acid" is one that is made by recombining nucleic acids, for example, during cloning, between DNA mixing or other methods; A "recombinant polypeptide" or "recombinant protein" is a polypeptide or protein that is produced by the expression of a recombinant nucleic acid. Examples of recombinant cells include cells that contain recombinant nucleic acids and / or recombinant polypeptides. The term "vector" refers to the means by which a nucleic acid can be propagated and / or transferred between organisms, cells or cellular components. Vectors include plasmids, viruses, bacteriophages, pro-viruses, phagemids, transposons, and artificial chromosomes and the like, which replicate autonomously or can be integrated on a chromosome of a host cell. A vector can also be a naked RNA polynucleotide, a naked DNA polynucleotide, a polynucleotide composed of both DNA and RNA within the same strand, a DNA or RNA conjugated with polylysine, a D? A or RA conjugated with peptide , a D? A conjugated with liposome or the like, which do not replicate autonomously. In the context of the invention, the term "isolated" refers to a biological material, such as a nucleic acid or a protein, that is substantially free of components that normally accompany or interact with it in its environment in which it occurs in a manner natural. The isolated material optionally comprises material not found with the material in its natural environment, for example, a cell. For example, if the material is in its natural environment, such as a cell, the material has been placed in a location in the cell (e.g., genome or genetic element) not native to a material found in that environment. For example, a nucleic acid that occurs naturally (eg, a coding sequence, a promoter, an enhancer, etc.) becomes isolated if it is introduced by means that does not occur naturally at a site in the genome. (eg, a vector, such as a plasmid or virus vector, or amplicon) non-native to that nucleic acid. A cell of Isolated plant, for example, can be in an environment (e.g., a cell culture system or purified from the cell culture) different from the native environment of the wild-type plant cells (e.g., an entire plant). The term "variant" with respect to a polypeptide refers to an amino acid sequence that is altered by one or more amino acids with respect to a reference sequence. The variant may have "conservative" changes, wherein a substituted amino acid has similar structural or chemical properties, for example, the replacement of leucine with isoleucine. Alternatively, a variant may have "non-conservative" changes, for example, replacement of a glycine with a tryptophan. The less analogous variation may also include the deletion or insertion of the amino acid, both. Guidance in determining which amino acid residues can be substituted, inserted or deleted without eliminating biological or immunological activity can be found using computer programs well known in the art, for example, DNASTAR software. Examples of conservative substitutions are also described below. A "host cell", as used herein, is a cell that has been transformed or transfected, or is capable of transformation or transfection, by an exogenous polynucleotide sequence. "Exogenous polynucleotide sequence" is defined to mean a sequence not naturally occurring in the cell, or that is naturally present in the cell but in a different genetic site, in a different copy number, or under the direction of a different regulatory element. A "promoter", as used herein, includes reference to a region of DNA upstream from the start of transcription and involved in the recognition and binding of RNA polymerase and other proteins to initiate transcription. A "plant promoter" is a promoter capable of initiating transcription in plant cells. Promoters of exemplary plants include, but are not limited to, those obtained from plants, plant viruses and bacteria comprising genes expressed in plant cells, such as Agrobacterium or Rhizobium. Examples of promoters under development control include promoters that preferentially initial transcription in certain tissues, such as leaves, roots or seeds or spatially in regions such as the endosperm, embryo or meristematic regions. Such promoters are referred to as "tissue-preferred" or tissue-specific. "A temporarily regulated promoter induces expression at particular times, such as between 0-25 days after pollination. cell "primarily induces expression in certain cell types in one or more organs, eg, vascular cells in roots or leaves.An" inducible "promoter is a promoter that is under environmental control and may be inducible or de-repressive. of environmental conditions that can affect transcription by inducible promoters include anaerobic conditions or the presence of light.The tissue-specific, cell-type and inducible promoters constitute the "non-constitutive" class of promoters.A "constitutive" promoter is a promoter that is active under most environmental conditions and in all or almost all tissues, and in all or almost all stages of development. "Transformation," as used herein, is the process by which a The cell is "transformed" by exogenous DNA when such exogenous DNA has been introduced into the cell membrane. it can not be integrated (covalently linked) into the chromosomal DNA that makes up the genome of the cell. In prokaryotes and yeasts, for example, exogenous DNA can be maintained on an episomal element, such as a plasmid. With respect to higher eukaryotic cells, a stably transformed or transfected cell is one in which exogenous DNA has been integrated into the chromosome so that it is inherited by daughter cells through the replication of the chromosome. This stability is demonstrated by the ability of the eukaryotic cell to establish cell lines or clones comprised of a daughter cell population containing the exogenous DNA. BRIEF DESCRIPTION OF THE DRAWINGS Figure 1 schematically illustrates the biosynthetic and ethylene signaling genes in plants, for example, Arabidopsis. Figure 2 schematically illustrates ACC synthase isolated and mapped genes and Mu insertion mutations. ACC6 is also known as ACS6, ACC2 is also known as ACS2, and ACC7 is also known as ACS7. Figure 3, Panels A, B, C, and D illustrate heterozygous ACC synthase inactivations in plants, e.g., corn. Panel A and Panel B illustrate heterozygous ACC synthase inactive plants in a field of wild-type plants. Panel C and Panel D illustrate leaves of an inactive heterozygous ACC synthase plant, left side of the panel, compared to the leaves of a wild-type ACC synthase plant, right side of the panel. Figure 4 illustrates an increased green retention attribute observed in leaves of plants that are inactive from homozygous ACC synthase (right) compared to wild type leaves (left) and inactive heterozygous leaves (middle part).

Figure 5, Panels A, B, C, D, E, and F illustrate the transpiration of the sheet (Panels A and D), stomatal conductance (Panels B and E) and assimilation of COc (Panels C and F) for sheets wild type (B73, + / +) and null mutants of ACS6 (15, 0/0) under control conditions (Panels A, B, and C) or drought conditions (Panels D, E, and F). For control conditions, the plants were cultivated under well-irrigated conditions and each leaf in a plant was measured in forty days after pollination (dap). For drought conditions, plants were grown under limited water conditions and each leaf on a plant was measured at forty days after pollination. The values represent an average of six determinations. Figure 6, Panels A, B and C illustrate sheet transpiration (Panel A), stomatal conductivity (Panel B) and COc assimilation (Panel C) for wild-type mutant leaves (B73, + / +), null of ACS2 (7, 0/0) and null of ACS6 (15, 0/0). The plants were grown under limited water conditions and each leaf on a plant was measured at forty days after pollination. The values represent an average of six determinations. Figure 7 illustrates schematically the phylogenetic analysis of the ACC synthase gene sequences, where the maize sequences are indicated by (A47 (also known as ACS2 or ACC2 herein), A50 (also known as ACS7 or ACC7 in the present), A65 (also known as ACS6 or ACC6 in the present)), the Arabidopsis sequences are indicated by (AtACS ...), the tomato sequences are indicated by (LeACS ...), the rice sequences are indicated by (indicates (OsiACS ...) and japonica (OsjACS ...)), wheat sequences are indicated by (TaACS ...) and the banana sequences are indicated by (MaACS .. .). Figure 8 illustrates a peptide consensus sequence alignment with the ACC synthase sequences of A47 (also known as ACS2 or ACC2), A50 (also known as ACS7 or ACC7) and A65 (also known as ACS6 or ACC6) with species both dicotyledonous (AtACS, LeACS) and monocotyledonous (OsiACS, OsjACS, TaACS and MaACS). The alignment is made with a very severe criterion (identical amino acids) and the plurality is 26.00, the threshold is 4, and the Average Weight is 1.00, the Average Equalization is 2.78 and the Average Unequal is -2.25. SEQ ID NOs are as follows: A47pep SEQ ID NO: 25; A50pep SEQ ID NO: 26; osiacslpep SEQ ID NO: 27; osjacslpep SEQ ID NO: 28; TaACS2pep SEQ ID NO: 29; AtACSlpep SEQ ID NO: 30; AtACS2pep SEQ ID NO: 31; LeACS2pep SEQ ID NO: 32; LeACS4pep SEQ ID NO: 33; MaACSlpep SEQ ID NO: 34; MaACS5pep SEQ ID NO: 35; LeACSIApep SEQ ID NO: 36; LeACSIBpep SEQ ID NO: 37; LeACSdpep SEQ ID NO: 38; AtACSdpep SEQ ID NO: 39; A65pep SEQ ID NO: 40; osiacs2pep SEQ ID NO: 41; AtACS5pep SEQ ID NO: 42; AtACS9 SEQ ID NO: 3; AtACS4pep SEQ ID NO: 44; AtACS8 SEQ ID NO: 45; MaACS2pep SEQ ID NO: 46; MaACS3pep SEQ ID NO: 47; LeACS3pep SEQ ID NO: 48; LeACS7pep SEQ ID NO: 9; OsjACS2pep SEQ ID NO: 50; OsjACS3 SEQ ID NO: 51; OsiACS3pep SEQ ID NO: 52; and AtACS7 SEQ ID NO: 53. Figure 9 illustrates a peptide consensus sequence alignment with the ACC47 synthase sequences of A47 (also known as ACS2 or ACC2), ASO (also known as ACS7 or ACC7) and A65 (also known as ACS6 or ACCd) with both dicotyledonous (AtACS, LeACS) and monocotyledonous species (OsiACS, OsjACS, TaACS, MaACS) ( SEQ ID NO: 25-53). The alignment is made with a criterion of severity (similar amino acid residues) and the plurality is 26.00, the threshold is 2, the Average Weight is 1.00, the Average Equalization is 2.78, and the Average Unequal is -2.25. Figure 10 illustrate a peptide consensus sequence alignment with the ACC synthase sequences of A47 (also known as ACS2 or ACC2), A50 (also known as ACS7 or ACC7) and A65 (also known as ACS6 or ACCd) with spices both dicotyledonous (AtACS, LeACS) and monocotyledonous (OsiACS, OsjACS, TaACS, MaACS) (SEQ ID NO: 25-53). The alignment is done with a less severe criterion (amino acid residues are few similar) and the plurality is 26.00, the threshold is 0, the Average Weight is 1.00, the Average Equalization is 2.78 and the Average Unequal is - 2. 25. Figure 11 illustrate a peptide consensus sequence alignment with the ACC47 synthase sequences of A47 (also known as ACS2 or ACC2) and A50 (also known as ACS7 or ACC7) with sequences that are very similar to ACS2 and ACS7 (SEQ ID NO: 25-39). The alignment is made with more severe criteria (identical amino acids) and the plurality is . 00, the threshold is 4, the Average Weight is 1.00, the Average Equalization is 2.78 and Average Disbalance is -2.25. Figure 12 illustrates a consensus sequence alignment of peptide with ACC sequences of A47 (also known as ACS2 or ACC2) and A50 (also known as ACS7 or ACC7) with sequences that are very similar to ACS2 and ACS7 (SEQ ID NO: 25-39). The alignment is made with severe criteria (similar amino acid residues) and the plurality is 15.00, the threshold is 2, the Average Weight is 1.00, the Average Equalization is 2.78 and the Average Unequalness is -2.25. Figure 13 illustrates a peptide consensus sequence alignment with A65 ACC synthase sequences (also known as ACS6 or ACCd) with sequences that are more similar to ACS6 (SEQ ID NO: 40-53). The alignment is made with more severe criteria (identical amino acids) and the plurality is 14.00, the threshold is 4, the Average Weight it is 1.00, the Average Equalization is 2.78 and the Average Unequal is -2.25. Figure 14 illustrates a peptide consensus sequence alignment with the ACC synthase sequences of A65 (also known as ACSd or ACCd) with sequences that are more similar to ACSd (SEQ ID NO: 40-53). The alignment is made with severe criteria (similar amino acid residues) and the plurality is 14.00, the threshold is 2, the Average Weight is 1.00, the Average Equalization is 2.78 and the Average Unequal is -2.25. Figure 15 illustrates a peptide consensus sequence alignment with ACC synthase sequences of A47 (also known as ACS2 or ACC2), A50 (also known as ACS7 or ACC7) and A65 (also known as ACS6 or ACCd) (SEQ ID NO. : 25-26 and 40). The alignment is made with more severe criteria (identical amino acids) and the plurality is 3.00, the threshold is 4, the Average Weight is 1.00, the Average Equalization is 2.78 and the Average Unequalness is -2.25. Figure 16 illustrates a peptide consensus sequence alignment with ACC synthase sequences of A47 (also known as ACS2 or ACC2), A50 (also known as ACS7 or ACC7) and A65 (also known as ACSd or ACCd) (SEQ ID NO. : 25-26 and 40). The alignment is made with severe criteria (similar amino acid residues) and plurality it is 3.00, the threshold is 2, the Average Weight is 1.00, the Average Equalization is 2.78 and the Average Unequal is -2.25. Figure 17 Panels A, B, C, and D illustrate the total chlorophyll data for inactive wild type and ACC synthase plants. Panels A and B illustrate the total chlorophyll data for wild type plants (B73, + / +), null of ACS2 (0/0) and null of ACSß (0/0) 40 days after pollination for plants grown under normal conditions (Panel A) or drought conditions (Panel B). Panel C compares total chlorophyll for wild type (B73, + / +) and null ACSd (0/0) plants 40 days after pollination under normal and drought conditions. Panel D illustrates a comparison of total chlorophyll for B73 (wild type) plants harvested 30-40 days after pollination. Figure 18 Panels A, B, C, and D illustrate soluble protein data for inactive wild type and ACC synthase plants. Panels A and B illustrate soluble protein data for wild type plants (B73, + / +), null of ACS2 (0/0) and null of ACS6 (0/0) 40 days after pollination for plants grown under normal conditions (Panel A) under drought conditions (Panel B). Panel C compares the soluble protein for wild-type (B73, + / +) and null ACS6 (0/0) plants 40 days after pollination under normal and drought conditions. Panel D illustrates a comparison of the soluble protein for B73 plants (wild type) collected in 30 and 40 days after pollination. Figure 19, Panels A and B illustrate the production of ethylene in seedling leaves. Panel A illustrates several lines. In Panel B, the seedling leaves are averaged by the genotype. In Panel C, ethylene production was determined for each wild type leaf (ie, B73) plants at 20, 30 and 40 DAP. Leaf 1 represents the oldest surviving leaf and leaf 11 the youngest. Three replicates were measured and the average and standard deviation are reported. Figure 20 Panel A illustrates the chlorophyll data, Panel B soluble protein and Panel C expression of Rubisco. The level of chlorophyll a + b (Panel A) soluble protein (Panel B) was measured on the third oldest leaf (Leaf 3), sixth oldest leaf (Sheet 6) and ninth oldest leaf (Sheet 9) of adult wild type plants (ie, ACS6 / ACS6), acs2 / acs2 and acs6 / acs6 after treatment in the dark for 7 days. The plants were irrigated daily. Additional acs6 / acs6 plants were irrigated daily with 100 μM ACC during the treatment. Acs6 / acsd sheets irrigated with 100 μM ACC but kept uncovered are also shown. The average deviation and Standard of the leaves of three individual plants is shown. (Panel C) Western analysis of the same leaves was performed using the anti-Rubisco rice antiserum. Soluble protein from leaf samples of equal fresh weight was used. Figure 21 Panels A-C illustrate the ACS2 fork construction. Panel A is a schematic diagram of PHP20600 containing a ubiquitin promoter (UBI1ZM PRO) that induces the expression of the ACS2 fork (a terminal repeat consisting of TR1 and TR2). RB represents the sequence of the right edge of Agrobacterium. A 49682 bp fragment of the cassette of 49682 bp is illustrated. Panel B presents the sequence of ZM-ACS2 TR1 (SEQ ID NO: 54) and Panel C presents the sequence of ZM-ACS2 TR2 (SEQ ID NO: 55). Figure 22 Panels A-C illustrate the ACS6 fork construction. Panel A is a schematic diagram of PHP20323 containing a ubiquitin promoter (UBI1ZM PRO) that induces the expression of the ACSd fork (a terminal repeat consisting of TR1 and TR2). RB represents the sequence of the right edge of Agrobacterium. A fragment of 3564 bp of cassette 49108 bp is illustrated. Panel B presents the sequence of ZM-ACS6 TR1 (SEQ ID NO: 56) and Panel C presents the sequence of ZM-ACSd TR2 (SEQ ID NO: 57). Figure 23 Panels A and B illustrate evidence generated for the ACS2 and ACS6 fork constructions.

Panel A presents a diagram that shows the number of individual evidences for the ACS2 fork (PHP20600) and the number of associated transgene copies by evidence. He Panel B presents a diagram that shows the number of individual evidences for the ACS6 fork (PHP20323) and the number of transgene copies associated by evidence. DETAILED DESCRIPTION "Green permanence" is a term commonly used to describe a plant phenotype. Green permanence is a desirable attribute in commercial agriculture, for example, a desirable attribute associated with grain filling. As described herein, five fundamentally different types of green permanence have been described, including Types A, B, C, D and E (see, for example, Thomas H and Smart CM (1993) Crops that stay green. of Applied Biology 123: 193-219, and Thomas H and Howarth CJ (2000) Five ways to stay green, Journal of Experimental Botany 51: 329-337). However, there is very little description of the biochemical, physiological or molecular basis for genetically determined green permanence. See, for example, Thomas and Howarth, supra. This invention provides a molecular / biochemical basis for green residence potential. A number of environmental and physiological conditions have been shown to significantly alter the synchronization and progression of the leaf's senescence and can provide some discernment in the basis for this attribute. Among environmental factors, light is probably the most significant, and it has been established for a long time that leaf senescence can be induced in many plant species by straining leaves in the dark. See, for example, Weaver LM and Amasino RM (2001) Senescence is induced in individual darkened Arabidopsis leaves, but inhibited in whole darkened plants. Plant Physiology 127: 876-886. The limited availability of nutrients and water has also been shown to induce senescence of the leaf prematurely. See, for example, Rosenow DT et al., (1983) Drought-tolerant sorghum and cotton genllplasm. Agricultural Water Management 7: 207-222. Among the physiological determinants, growth regulators play a key role in directing the leaf's senescence program. The modification of cytokinin levels can significantly slow the leaf's senescence. For example, plants transformed with isopentenyl transferase (ipt), a gene from Agrobacterium um that encodes a limiting step of the ratio in cytokinin biosynthesis, when placed under the control of an inducible senescence promoter, resulted in a self-regulated cytokinin production and a strong green permanence phenotype. See, for example, Gan S and Amasino RM (1995) Inhibition of leaf senescence by autoregulated production of cytokinin. Science 270: 1986-1988. However, there are other factors that are involved with this attribute. For example, ethylene has also been implicated in the control of leaf senescence (see, for example, Davis KM and Grierson D (1989) Identification of cDNA clones for tomato (Lycopersicon esculentum Mili.) MRNAs that accumulate during fruit collection and leaf senescence in response to ethylene. Plant 179: 73-80) and some dicotyledonous plants impaired in the production or perception of ethylene also show a delay in leaf senescence (see, for example, Picton S et al., (1993) Al teredfrui t ripeníng and leaf senescence ifz tomatoes exprés sing an antisense ethylene-forming enzyme transgene The Plant Journal 3: 469-481; Grbic V and Bleeker AB (1995) Ethylene regulates the timing of leaf senescence in Arabidopsis, The Plant Journal 8: 95-102; John I et al., (1995) Delayed leaf senescence in ethylene-deficient ACC-oxidase antisense tomato plants: molecular and physiological analysis, The Plant Journal 7: 483-490), which can be phenotyped by the exogenous application of biosynthesis inhibitors and ethylene action (see, for example, Abeles FB et al., (1992) Ethylene in Plant Biology, Academic Press, San Diego, CA).

The perception of ethylene involves receptors located in the membrane that, for example, in Arabidopsis include ETR1, ERS1, ETR2, ERS2 and EIN4 (see Figure 1). ETR1, ETR2 and EIN4 are composed of three domains, an N-terminal ethylene binding domain, a putative histidine protein kinase domain and a C-terminal receptor domain, while ERS1 and ERS2 lack the receptor domain. These genes have been grouped into two subfamilies based on homology, where ETR1 and ERS1 comprise one subfamily and ETR2, ERS2 and EIN4 comprise the other. In Arabidopsis, the analysis of the loss of function of the mutants has revealed that ethylene inhibits the signaling activity of these receptors and subsequently their ability to activate CTR1, a negative regulator of ethylene responses that is related to serine / threonine kinases of RAF type mammals. The ethylene signal transduction pathway suggests that the ethylene bond to the receptor inhibits its own kinase activity, resulting in decreased activity in CTR1, and consequently, an increase in the activity of EIN2 (which acts downstream of CTR1) finally it leads to increases in ethylene responsiveness. The differential expression of the members of the ethylene receptor family has been observed, both in development and in response to ethylene.

The identification and analysis of mutants in Arabidopsis and tomato that are deficient in the biosynthesis and perception of ethylene are valuable in establishing the role that ethylene plays in the growth and development of the plant. The analysis of the mutant has also been instrumental in identifying and characterizing the ethylene signal transduction pathway. While many ethylene mutants have been identified in dicotyledonous plants (eg, Arabidopsis and tomato), none of these mutants have been identified in monocots. (for example, rice, wheat and corn). Here, for example, ethylene mutants (e.g., in a monocot) deficient in ACC synthase, the first synthesis in the ethylene biosynthetic pathway, are described. This invention provides ACC synthase polynucleotide sequences from plants, which modulate the green residence potential in plants and the production of ethylene, exemplified by, for example, SEQ ID NO: 1 to SEQ ID NO: 6 and SEQ ID NO. : 10 and, for example, a set of polypeptide sequences that modulate the green residence potential in plants and / or the production of ethylene, eg, SEQ ID NOY to SEQ ID NO: 9 and SEQ ID NO: 11. invention also provides inactive plant cells deficient in ACC synthase in inactive plants that have a green permanence potential phenotype, as well as inactive plants that have a male sterility phenotype. The plants of the invention may have the characteristic of regulating the responses to environmental stress better than the control plants, for example, higher tolerance to stress by drought. The plants of the invention may also have a higher tolerance for other locations (e.g., settling in, for example, corn) compared to the control plants. Thus, the plants of the invention can be planted in densities higher than what is currently practiced by farmers. In addition, the plants of the invention are critical in clarifying the regulatory functions that the ethylene performs throughout the development of the plant as well as its function in regulating the responses to stress, for example, drought, stacking, etc. ETHYLENE IN PLANTS Ethylene (C_H4) is a gaseous plant hormone. This has a varied spectrum of effects that may be specific to the tissue and / or the species. For example, physiological activities include, but are not limited to, promotion of the maturation of the food, abscission of leaves and fruits of dicotyledonous species, senescence of the flower, extension of the stem of aquatic plants, development of the gaseous space (aerenchyma) in roots, epinásticas curvatures of the leaf, swelling of the stem and shoot (in association with atrophy), female ability in curcubitáceas, growth of fruit in certain species, closure of the apical hook in etiolate shoots, formation of small roots, flowering in the Bromeliaceae, diageotropism of etiolate shoots and increased gene expression (for example, polygalacturonase, cellulase, chitinases, β-1, 3-glucanases, etc.). Ethylene is released naturally as the fruit ripens and is also produced by most plant tissues, for example, in response to stress (eg, drought, settling, disease or pathogen attack, temperature stress (hot or cold) ), injury, etc.), and in the maturation and senescence of the organs. Ethylene is generated from methionine by a well-defined route involving the conversion of S-adenosyl-L-methionine (SAM or Aso Met)) or the cyclic amino acid a 1-aminociclopropane-l-carboxylic acid (ACC) that it is facilitated by the ACC synthase (see, for example, Figure 1). Sulfur is conserved in the process by recycling 5'-metithioadenosine. The ACC synthase is an aminotransferase that catalyzes the limiting step of proportion in ethylene formation by converting S-adenosylmethionine to ACC. Typically, the enzyme requires pyridoxal phosphate as a cofactor. The ACC synthase is typically encoded in multigenes families.

Examples include SEQ ID NO: 1-3, described herein. Individual members may exhibit specific tissue regulation and / or be induced in response to environmental and chemical stimuli. The features of the invention include sequences and subsequences of ACC synthase. See the section entitled "Polynucleotides and Polypeptides of the Invention". Ethylene is then produced from the oxidation of ACC through the action of ACC oxidase (also known as the ethylene-forming enzyme) with hydrogen cyanide as a by-product that is detoxified by β-cyano-alanine synthase. ACC oxidase is encoded by families of multigenes in which individual members exhibit specific tissue regulation and / or are induced in response to environmental and chemical stimuli. The activity of ACC oxidase can be inhibited by anoxia and cobalt ions. The enzyme ACC oxidase is stereospecific and uses cofactors, for example, Fe + 2, O? Ascorbate, etc. Finally, ethylene is metabolized by oxidation to CO 2 or ethylene oxide and ethylene glycol. POLINUCLEOTIDES AND POLIPEPTIDES OF THE INVENTION The invention characterizes the identification of gene sequences, nucleic acid coding sequences and amino acid sequences of ACC synthase, which are associated, for example, with the permanency potential green in plants and / or the production of ethylene. The sequences of the invention can influence the green staying potential in plants by modulating the production of ethylene. The polynucleotide sequences of the invention include, for example, the polynucleotide sequences represented by SEQ ID NO: 1 to SEQ ID NO: 6 and SEQ ID NO: 10 and subsequences thereof. In addition to the sequences expressly provided in the accompanying sequence listing, the invention includes polynucleotide sequences that are highly structurally and / or functionally related. For example, polynucleotides that encode polypeptide sequences represented by SEQ ID NOY to SEQ ID NO: 9 and SEQ ID NO: 11 or subsequences thereof, are one embodiment of the invention. In addition, the polynucleotide sequences of the invention include polynucleotide sequences that hybridize under severe conditions to a polynucleotide sequence comprising any of SEQ ID NO: l-SEQ ID NO: 6 and SEQ ID NO: 10 or a subsequence of the same (for example, a subsequence comprising at least 100 contiguous nucleotides). The polynucleotides of the invention also include sequences and / or subsequences of ACC synthase configured for the production of RNA, for example, mRNA, antisense RNA, sense RNA, configurations of silencing or RNA interference, etc. In addition to the polynucleotide sequences of the invention, for example, listed in SEQ ID NO: 1 to SEQ ID NO: 6 and SEQ ID NO: 10, polynucleotide sequences that are substantially identical to a polynucleotide of the invention can to be used in the compositions and methods of the invention. Substantially identical or substantially similar polynucleotide sequences are defined as polynucleotide sequences that are identical, on a nucleotide-by-nucleotide basis, with at least one subsequence of a reference polynucleotide, for example, selected from SEQ ID NO: 1 -6 and 10. Such polynucleotides can include, for example, insertions, deletions and substitutions relative to any of SEQ ID NO: 1-6 and 10. For example, such polynucleotides are typically at least about 70% identical to a reference polynucleotide selected from SEQ ID NO: 1 to SEQ ID NO: 6 and SEQ ID NO: 10 or subsequences thereof. For example, at least 7 out of 10 nucleotides within a comparison window are identical to the reference sequence selected, for example, from SEQ ID NO: 1-6 and 10. Frequently, such sequences are at least about 70%, at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 98%, at least about 99% or at least about 99.5%, identical to the reference sequence, eg, at least one of SEQ ID NO: SEQ ID NO: 6 or SEQ ID NO: 10. The subsequences of the polynucleotides of the invention described above, for example, SEQ ID NO: 1-6 and 10, including, for example, at least about 5, at least about 10, at least about 15, at least about 20, at least about 25, at least about 50, at least about 75, at least about 100, at least about 500, about 1000 or more, nucleotides contiguous or complementary subsequences thereof are also a feature of the invention. Such subsequences can be, for example, oligonucleotides, such as synthetic oligonucleotides, isolated oligonucleotides or full-length genes or cDNAs. In addition, polynucleotide sequences complementary to any of the sequences described above are included among the polynucleotides of the invention. The polypeptide sequences of the invention include, for example, the amino acid sequences - represented by SEQ ID NOY to SEQ ID NO: 9 and SEQ ID _ > / NO: 11 and subsequences thereof. In addition to the sequences expressly provided in the accompanying sequence listing, the invention includes amino acid sequences that are highly related in a structural and / or functional manner. For example, in addition to the amino acid sequences of the invention, for example, listed in SEQ ID NOY to SEQ ID NO: 9 and SEQ ID NO: 11, amino acid sequences that are substantially identical to a polypeptide of the invention can be used in the compositions and methods of the invention. Substantially identical or substantially similar amino acid sequences are defined as amino acid sequences that are identical, on an amino acid-by-amino acid basis, with at least one subsequence of a reference polypeptide, for example, selected from SEQ ID NO: 7 -9 and 11. Such polypeptides may include, for example, insertions, deletions and substitutions relative to any of SEQ ID NO: 7-9 and 11. For example, such polypeptides are typically at least about 70% identical to a reference polypeptide selected from SEQ ID NOY to SEQ ID NO: 9 and SEQ ID NO: 11 or a subsequence thereof. For example, at least 7 out of 10 amino acids within a comparison window are identical to the selected reference sequence, for example, of SEQ ID NO: 7-9 and 11. Frequently, such sequences are at least about 70%, at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 98% , at least about 99% or at least about 99.5%, identical to the reference sequence, for example, at least one of SEQ ID NOY to SEQ ID NO: 9 or SEQ ID NO: 11. The subsequences of the polypeptides of the invention described above, for example, SEQ ID NO: 7-9 and 11, including, for example, at least about 5, at least about 10, at least about 15, at least about 20, at least about 25, at least about 50, at least about 75, at least about 100, at least about 500, about 1000 or more, contiguous amino acids are also a feature of the invention. The conservative variants of amino acid sequences or subsequences of the invention are also amino acid subsequences of the invention. The polypeptides of the invention are optionally immunogenic, enzymatically active, enzymatically inactive and the like. Where the polynucleotide sequences of the invention are translated to form a polypeptide or subsequence of a polypeptide, the nucleotide changes they can result in amino acid substitutions, either conservative or non-conservative. Conservative amino acid substitutions refer to the interchangeability of residues having functionally similar side chains. Conservative substitution tables that provide functionally similar amino acids are well known in the art. Table 1 shows six groups containing amino acids that are "conservative substitutions" with each other. Other conservative substitution tables are available in the art and can be used in a similar manner. Table 1: Conservative Substitution Groups 1 Alanine (A) Serine (s) Threonine (T) 2 Aspartic Acid (D) Glutamic Acid (E) 3 Asparagine (N) Glutamine (Q) 4 Arginine (R) Lysine (K) 5 Isoleucine (I) Leucine (L) Methionine (M) Valine (V) 6 Phenylalanine (F) Tyrosine (Y) Tryptophan (W) One skilled in the art will appreciate that many conservative substitutions of nucleic acid constructs that are disclosed produce functionally identical constructs. For example, as discussed in the above, due to the degeneracy of the genetic code, "silent substitutions" (that is, substitutions in a nucleic acid sequence that do not result in an alteration in a coded polypeptide) are an implied feature of each nucleic acid sequence encoding an amino acid. Similarly, "conservative amino acid substitutions" or a few amino acids in an amino acid sequence (eg, about 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9% , 10% or more) are substituted with amino acids with highly similar properties, they are also easily identified as being highly similar to a disclosed construction. Such conservative variations of each disclosed sequence are a feature of the invention. Methods for obtaining conservative variants, as well as more divergent versions of the nucleic acids and polypeptides of the invention, are widely known in the art. In addition to the naturally occurring homologues that can be obtained, for example, by classifying genomic or expression libraries according to any of a variety of well-established protocols, see, for example, Ausubel et al., Current Protocols in Molecular Biology. (supplemented through 2004) John Wiley & Sons, New York ("Ausubel"); Sambret al., Molecular Cloning-A Laboratory Manual (2nd Edition), Volume 1-3, Cold Spring Harbor Laboratory, Cold Spring Harbor, New York, 1989 ("Sambr) and Berger and Kimmel Guide to Molecular Cloning Techniques, Methods in Enzymology volume 152 Academic Press, Inc., San Diego, CA ("Berger"), additional variants can be produced by any of a variety of mutagenesis procedures. Many such methods are known in the art, including site-directed mutagenesis, oligonucleotide-directed mutagenesis, and many others. For example, site-directed mutagenesis is described, for example, in Smith (1985) "T n vi tro mutagenesis" Ann. Revolution. Genet 19: 423-462 and references therein, Botstein & Shortle (1985) "for example, in Botstein &Shortle (1985)" Strategies and applications of in vitro mutagenesis "Science 229: 1193-1201; and Carter (1986)" Si te-directed mutagenesis "Biochem. J. 237 : 1-7: Oligonucleotide-directed mutagenesis is described, for example, in Zoller &Smith (1982) "Oligonucleotide-directed mutagenesis using MI 3-derived vectors: an efficient and general procedure for the production ofpoiti t utations in any DNA f agment "Nucleic Acids Res. 10: 6487-6500.) Mutagenesis using modified bases is described, for example, in Kunkel (1985)" Rapid and efficient if te-specific mutagenesis wi thout phenotypic selection "Proc. Nati. Acad. Sci. USA 82: 488-492 and Taylor et al., (1985) "The rapxd generation of oligofaucleotide-directed mutations at the high frequency using phosphorothioate-modified DNA" Nucí Acids Res. 13: 8765-8787.

Mutagenesis using spaced duplex DNA is described, for example, in Kramer et al., (1984) "The gapped duplex DNA approach to oligonucleotide-directed mutation construction" Nucí. Acids Res. 12: 9441-9460). Mutagenesis of dot mismatch is described, for example, by Kramer et al., (1984) "PointMismatch Repair" Cell 38: 879-887). Double-strand break mutagenesis is described, for example, in Mandecki (1986) "Oligonucleotide-directed double-strand break repair in plasmids of Escherichia coli: a methodfox site-specific mutagenesis" Proc. Nati Acad. Sci. USA, 83: 7177-7181, and Arnold (1993) "Protein engineering for unusual environments" Current Opinion in Biotechnology 4: 450-455). Mutagenesis using defective host strains of repair is described, for example, in Carter et al. (1985) "Improved oligonucleotide site-directed fnutageyiesis using M13 vectors" Nucí. Acids Res. 13: 4431-4443. Mutagenesis by total gene synthesis is described, for example, by Nambiar et al., (1984) "Total synthesis and cloning of a gene coding for the ribonuclease S protein" Science 223: 1299-1301. The intermixing of DN is described, for example, by Steer (1994) "Rapid evolution of a protein in vitro by DNA shuffling" Nature 370: 389-391 and Stemmer (1994) "DNA shuffling by random fragmentation and reassembly: In vi tro recombination for molecular evolution. " Proc. Nati Acad. Sci. USA 91: 10747-10751. Many of the above methods are also described in for example, Methods in Enzymology Volume 154, which also describes useful controls for repair problems with various methods of mutagenesis. Equipment for mutagenesis, library construction and other diversity generation methods are also commercially available. For example, equipment is available from, for example, Amersham International foot (Piscataway, NJ) (for example, using the previous Eckstein method), Bio / Can Scientific (Mississauga, Ontario, CANTADA), Bio-Rad (Hercules, CA) (for example, using the Kunkel method described above), Boehringer Mannheim Corp. (Ridgefield, Connecticut), Clonetech Laboratories of BD Biosciences (Palo Alto, CA), DNA Technologies (Gaithersburg, MD), Epicenter Technologies ( Madison, Wl) (for example, the 5 bonus 3 bonus team); Genpak Inc. (Stony Brook, New York), Lemargo Inc (Toronto, CANADA), Invitrogen Life Technologies (Carisbad, California), New England Biolabs (Beverly, MA), Pharmacia Biotech (Peapack, New Jersey), Promega Corp. ( Madison, Wl), QBiogene (Carisbad, CA) and Estratagene (La Jolla, CA) (eg empl o, QuickChange ™ site-directed mutagenesis team and Chameleon ™ double-stranded site-directed mutagenesis team). Determination of Relationships of Sequences The nucleic acid and amino acid sequences of the invention include, for example, those provided in SEQ ID NO: 1 to SEQ ID NO: 11 and subsequences thereof, as well as similar sequences. The sequences similar are objectively determined by any number of methods, for example, percent identity, hybridization, immunologically and the like. A variety of methods for determining relationships between two or more sequences (eg, identity, similarity and / or homology) are available and well known in the art. The methods include manual alignment, computer assisted sequence alignment and combinations thereof, for example. A number of algorithms (which are generally implemented on a computer) to perform sequence alignment are widely available or can be produced by an expert. These methods include, for example, the local homology algorithm of Smith and Waterman (1981) Adv. Appl. Math. 2: 482; the homology alignment algorithm of Needleman and Wunsch (1970) J. Mol. Biol. 48: 443; the search for the similarity method of Pearson and Lipman (1988) Proc. Nati Acad. Sci. (USA) 85: 2444; and / or through co-automated implementations of these algorithms (for example, GAP, BESTFIT, FASTA and TFASTA in the Wisconsin Genetics Software Package Résease 7.0, Genetics Computer Group, 575 Science Dr., Madison, Wl).

For example, software for conducting sequence identity analysis (sequence similarity) using the BLAST algorithm is described in Altschul et al., (1990) J. Mol. Biol. 215: 403-410. This software is publicly available, for example, through the National Center for Biotechnology Information on the global network at ncbi.nlm.nih.gov. This algorithm first involves identifying high register sequence pairs (HSPs) by identifying short words of length W in the query sequence, which either equal or satisfies some positive value threshold register T when it is deleted with a word of the same value. length in a database sequence. T is referred to as the close word registration threshold. These initial near record hits act as seeds to initiate searches to find longer HSPs that contain them. Word hits are then extended in both directions along each sequence while the cumulative alignment record can be incremented. Cumulative records are calculated using, for nucleotide sequences, the parameters M (numbering register for an equalization residuals, always> 0) and N (registration of the residual designation sanction, always <0). For amino acid sequences, a register matrix is used to calculate cumulative records. The extension of the word hits in each direction are t > ts interrupted when: the cumulative alignment record descends by the amount X of its maximum achieved value; the cumulative record goes from zero or below, due to the accumulation of one or more negative record residue alignment; or the end of any sequence is reached. The parameters of the BLAST algorithm W, T and X determine the sensitivity and speed of the alignment. The BLASTTN program (for nucleotide sequences) uses as errors a word length (W) of 11, an expectation (E) of 10, a cut of 100, M = 5, N = -4 and a comparison of both strands. For amino acid sequences, the BLASTTP program (BLAST Protein) uses as errors a word length (W) of 3, an expectation (E) of 10 and the registration matrix BLOSUM62 (see, Henikoff &Henikoff (1989) Proc. Nati, Acad. Sci. USA 89: 10915). Additionally, the BLAST algorithm performs a statistical analysis of the similarity between two sequences (see, for example, Karlin &Altschul (1993) Proc. NatY.

Acad. Sci. USA 90: 5873-5787). A measure of similarity provided by the BLAST algorithm is the probability of smaller sums (p (N)), which provides an indication of the probability by which an equalization between two nucleotide or amino acid sequences would occur by chance. For example, a nucleic acid is considered similar to a reference sequence (and, therefore, in this context, homologous) if the probability of smaller sums in a comparison of the test nucleic acid to the reference nucleic acid is less than about 0.1 or less than about 0.01 or even less than about 0.001. Another example of a useful sequence alignment algorithm is PILEUP. PILEUP creates a multiple sequence alignment from a group of related sequences using alignments by progressive pairs. It can also graph a tree that shows the grouping relationships used to create the alignment. PILEUP uses a simplification of the progressive alignment method Feng & Doolittle (1987) J. Mol. Evol. 35: 351-360. The method used is similar to the method described by Higgins & Acute (1989) CABI0S5: 151-153. The program can align, for example, up to 300 sequences of maximum length of 5,000 letters. The multiple alignment procedure begins with the pairwise alignment of the two most similar sequences, producing a grouping of two aligned sequences. This grouping can then be aligned to the next most related sequence or grouping of aligned sequences. Two groupings of sequences can be aligned by a simple extension of the pairwise alignment of two individual sequences. The final alignment is achieved through a series of alignments by progressive pairs. The program also it can be used to graph a dendrogram or tree representation of grouping relationships. The program is run by designating specific sequences and their amino acid or nucleotide coordinates for sequence comparison regions. A further example of an algorithm that is suitable for multiple DNA, or amino acid, or sequence alignments is the CLUSTALW program (Thompson, J. D. et al., (1994) Nucí, Acids, Res. 22: 4673-4680). CLUSTALW performs multiple pair comparisons between groups of sequences and assembles them into a multiple alignment based on homology. The penalties of space opening and space extension can be, for example, 10 and 0.05 respectively. For amino acid alignments, the BLOSUM algorithm can be used as a protein weight matrix. See, for example, Henikoff and Henikoff (1992) Proc. Nati Acad. Sci. USA 89: 10915-10919. Hybridization of Nucleic Acid The similarity between the ACC synthase nucleic acids of the invention can also be evaluated by "hybridization" between single-stranded nucleic acids (or single-stranded regions) with complementary or partially complementary polynucleotide sequences. Hybridization is a measure of the physical association between nucleic acids, typically in solution, or with one of the nucleic acid strands immobilized on a solid support, for example, a membrane, a bead, a chip, a filter, etc. Nucleic acid hybridization occurs based on a variety of well-characterized physiochemical forces, such as hydrogen bonding, solvent exclusion, base stacking and the like. Numerous protocols for nucleic acid hybridization are well known in the art. An extensive guide for nucleic acid hybridization is found in Tijssen (1993) Laboratory Techniques in Biochemistry and Molecular Biology - Hybridization with Nucleic Acid Probes, Part I, Chapter 2, "Overview of Principles of Hybridization and the Strategy of Nucleic Acid Test assays ", (Elsevier, New York), as well as in Ausubel et al., Current Protocols in Molecular Biology (supplemented until 2004) John Wiley & Sons, New York ("Ausubel"); Sambrook et al., Molecular Cloning-A Laboratory Manual (2- Edition), Volume 1-3, Cold Spring Harbor Laboratory, Cold Spring Harbor, New York, 1989 ("Sambrook") and Berger and Kimmel Guide to Molecular Cloning Techniques, Methods in Enzymology volume 152 Academic Press, Inc., San Diego, CA ("Berger"). Hames and Higgins (1995) Gene Probes 1. IRL Press at Oxford University Press, Oxford, England (Hames and Higgins 1) and Hames and Higgins (1995) Gene Probes 2, IRL Press at Oxford University Press, Oxford, England (Hames and Higgins 2) provide details on the synthesis, marking, detection and quantification of DNA and RNA, including oligonucleotides. The appropriate conditions to obtain hybridization, including differential hybridization, are selected according to the theoretical melting temperature (Tm) between complementary and partially complementary nucleic acids. Under a given set of conditions, for example, solvent composition, ionic concentration, etc., the Tm is the temperature at which the duplex between the hybridizing nucleic acid strands is 50% denatured. That is, the Tm corresponds to the temperature corresponding to the midpoint in the transition from the helix to the random spiral; this depends on the length of the polynucleotides, nucleotide composition and ion concentration, for long nucleotide stretches. After hybridization, unhybridized nucleic acids can be removed by a series of washes, the severity of which can be adjusted depending on the desired results. Low severity wash conditions (eg, using higher salt and lower temperature) increase sensitivity, but may produce non-specific hybridization signals and high background signals. The conditions of highest severity (for example, using lower salt and higher temperature which is closer to the Tm) the antecedent signal decreases, typically with mainly the remaining specific signal. See, also, Rapley, R. and Walker, J.M. eds., Molecular Biomethods Handbook (Humana Press, Inc. 1998). "Severe hybridization wash conditions" or "severe conditions" in the context of nucleic acid hybridization experiments, such as Southern and Northern hybridizations, are sequence dependent, and are different under different environmental parameters. An extensive guide for nucleic acid hybridization is found in Tijssen (1993), supra, and in Hames and Higgins 1 and Hames and Higgins 2, supra. An example of severe hybridization conditions for the hybridization of complementary nucleic acids having more than 100 complementary residues on a filter in a Southern or northern spotting is 2x SSC, 50% formamide at 42 ° C, with hybridization being carried performed overnight (for example, for approximately 20 hours). An example of severe washing conditions is a 0.2x SSC wash at 65 ° C for 15 minutes. { see Sambrook, supra for a description of the SSC regulatory solution). Frequently, the wash that determines the severity is preceded by a low severity wash to remove the signal due to the unhybridized residual probe. An example of low severity wash is 2x SSC at room temperature (eg, 20 ° C during 15 minutes) . In general, a signal-to-noise ratio of at least 2.5x-5x (and typically higher) than that observed for an unrelated probe in the particular hybridization assay indicates the detection of a specific hybridization. The detection of at least severe hybridization between two sequences in the context of the invention indicates the relatively strong structural similarity to, for example, the ACC synthase nucleic acids provided in the sequence listings herein. For purposes of the invention, generally, "highly-severe" hybridization and washing conditions are selected to be about 5 ° C or less lower than the thermal melting point (Tm for the specific sequence at a defined ionic concentration and pH (as is mentioned below, highly severe conditions can also be referred to in comparative terms.) Target sequences that are closely related or identical to the nucleotide sequence of interest (eg, "probe") can be identified under severe or highly The conditions of lesser severity are appropriate for sequences that are less complementary, for example, in the determination of severe or highly severe hybridization (or even more severe hybridization) and washing conditions, the severity of the hybridization and the washing conditions is gradually increased (for example, by increasing the temperature, decreasing the salt concentration, increasing the detergent concentration and / or increasing the concentration of the organic solvents, such as formamide, in hybridization or washing), until a selected set of criteria is met. For example, the severity of hybridization and washing conditions is gradually increased until a probe comprises one or more polynucleotide sequences of the invention, for example, selected from SEQ ID NO: SEQ ID NO: 6 and SEQ ID NO: 10 or a subsequence thereof and / or polynucleotide sequences complementary thereto, binds to a perfectly matched complementary target (again, as a nucleic acid comprising one or more selected nucleic acid sequences or subsequences of SEQ ID NO. N0: 1 to SEQ ID NO: 6 and SEQ ID NO: 10 and complementary polynucleotide sequences thereof), with a signal to noise ratio that is at least 2.5x, and optionally 5x or lOx or lOOx or greater, as high as that observed for the hybridization of the probe to an unmatched target, as desired. Using subsequences derived from the nucleic acids encoding the ACC synthase polypeptides of the invention, objective nucleic acids can be obtained novelties; such target nucleic acids are also a feature of the invention. For example, such objective nucleic acids include sequences that hybridize under severe conditions to an oligonucleotide probe that corresponds to a unique subsequence of any of the polynucleotides of the invention, for example, SEQ ID NO: 1-6, 10 or a complementary sequence from it; the probe optionally encodes a unique subsequence in any of the polypeptides of the invention, for example, SEQ ID NO: 7-9 and 11. For example, hybridization conditions are selected under which an objective oligonucleotide is perfectly complementary to the probe of hybrid oligonucleotide to the probe with at least about a 5-10x higher signal-to-noise ratio than for hybridization of the target oligonucleotide to a non-complementary, negative control nucleic acid. Higher signal-to-noise ratios can be achieved by increasing the severity of hybridization conditions such that ratios of approximately 15x, 20x, 30x, 50x or greater are obtained. The particular signal will depend on the brand used in the relevant assay, for example, a fluorescent label, a colorimetric label, a radioactive label or the like. Vectors, Promoters and Expression Systems The nucleic acids of the invention can be in any of a variety of forms, for example, expression cassettes, vectors, plasmids or linear nucleic acid sequences. For example, vectors, plasmids, cosides, bacterial artificial chromosomes (BACs), YACs (yeast artificial chromosomes), phage, viruses and nucleic acid segments may comprise a nucleic acid sequence of ACC synthase or subsequence thereof. which one you want to enter into the cells. These nucleic acid constructs may also include promoters, enhancers, polylinkers, regulatory genes, etc. Thus, the present invention also relates, for example, to vectors comprising the polynucleotides of the present invention, host cells that incorporate the vectors of the invention and the production of polypeptides of the invention by recombinant techniques. According to this aspect of the invention, the vector can be, for example, a plasmid vector, a single or double-stranded phage vector or a single or double-stranded RNA or DNA viral vector. Such vectors can be introduced into cells as polynucleotides, preferably DNA, by well known techniques for introducing DNA and RNA into cells. Vectors, in the case of phage and viral vectors, can also be and Preferably they are introduced into cells as packaged or encapsidated viruses by well-known techniques for infection and transduction. Viral vectors may be replication competent or replication defective. In the latter case, viral propagation will generally only occur in the complement of host cells. Among the preferred vectors, in certain aspects, are those for expression of polynucleotides and polypeptides of the present invention. Generally, such vectors comprise cis action control regions effective for host expression, operably linked to the polynucleotide that is expressed. Appropriate trans action factors are supplied by host, supplied by a complementation vector or supplied by the vector itself at introduction to the host. In certain preferred embodiments in this regard, the vectors provide the preferred expression. Such a preferred expression may be inducible expression, temporally limited expression, or expression restricted to certain cell types predominantly, or any combination of the previous ones. Particularly preferred inducible vectors are vectors that can be induced for expression by environmental factors that are easy to manipulate, such as temperature and nutrient additives.

A variety of vectors suitable for this aspect of the invention, including constitutive and inducible expression vectors for use in prokaryotic and eukaryotic hosts, are well known and routinely employed by those skilled in the art. Such vectors include, but are not limited to, chromosomal, episomal and virus derivatives, eg, vectors derived from bacterial plasmids, bacteriophage plasmids, transposons, yeast episomes, insertion elements, yeast chromosomal elements, virus such as baculovirus, papova virus, such as SV40, vaccinia virus, adenovirus, fowl pox virus, pseudorabies virus and retroviruses, and vectors derived from combinations thereof, such as those derived from plasmid and bacteriophage gene elements such as phagemid cosmetics and binaries used for transformations mediated by Agrobacterium. Everything can be used for the expression according to this aspect of the present invention. Vectors that are functional in plants can be binary plasmids derived from Agrobacterium. Such vectors are capable of transforming plant cells. These vectors contain left and right border sequences that are required for integration into the host (plant) chromosome. At a minimum, between these border sequences is the gene (or other polynucleotide sequence of the present invention) which is expressed, typically under the control of regulatory elements. In one modality, a selectable marker and a reporter gene are also included. For ease of obtaining sufficient amounts of the vector, a bacterial origin that allows replication in E. coli can be used. The following vectors, which are commercially available, are provided by way of example. Among Iso vectors preferred for use in bacteria are pQE70, pQE60 and pQE-9, available from Qiagen; pBS vectors, Phagescript vectors, Bluescript vectors, pNH8A, pNHlda, pNH18A, pNH46A, available from Stratagene; and ptrc99a, pKK223-3, pKK233-3, pDR540, pRIT5 available from Pharmacia. Among the preferred eukaryotic vectors are pWLNEO, pSV2CAT, pOG44, pXT1 and pSG available from Stratagene; and pSVK3, pBPV, pMSG and pSVL available from Pharmacia. Binary vectors useful plants include BIN19 and its available Clontech derivatives. These vectors are listed only by way of illustration of many of the commercially available and well known vectors that are available to those skilled in the art for use in accordance with this aspect of the present invention. It will be appreciated that any other plasmid or vector suitable for, for example, the introduction, maintenance, propagation or expression of a polynucleotide or polypeptide of the invention in a host they can be used in this aspect of the invention, several of which are disclosed in more detail below. In general, the expression constructs will contain sites for transcription initiation and termination, and, in the transcribed region, a ribosome binding site for translation when the construct encodes a polypeptide. The coding portion of the mature transcripts expressed by the constructs will include a translation initiation AUG at the start and a termination codon appropriately located at the end of the polypeptide that is translated. In addition, the constructions may contain control regions that regulate as well as engender the expression. Generally, according to many commonly practiced procedures, such regions will operate by controlling transcription, such as transcription factors, repressor binding sites and termination signals, among others. For the secretion of a translated protein in the lumen of the endoplasmic reticulum, in the periplasmic space or in the extracellular environment, appropriate secretion signals can be incorporated into the expressed polypeptide. These signals may be endogenous to the polypeptide or may be heterologous signals. The transcription of the DNA (e.g., encoding the polypeptides) of the present invention by Higher eukaryotes can be increased by inserting an enhancer sequence into the vector. Enhancers are cis-acting elements of DNA, usually approximately 10 to 300 bp, which act to increase the transcriptional activity of a promoter in a given host cell type. Examples of enhancers include the SV40 enhancer, which is located on the late side of the replication origin at bp 100 to 270, the cytomegalovirus early promoter enhancer, the polyoma enhancer on the late side of the replication origin, and adenovirus enhancers.

Additional enhancers useful in the invention for increasing transcription of the introduced DNA segment, include, inter alia, similar viral enhancers those within the 35S promoter, as shown by Odell et al., Plant Mol. Biol. 10: 263-72 (1988) and an opine gene enhancer as described by Fromm et al., Plant Cell 1: 977 (1989). The enhancer can affect the specificity of the tissue and / or the temporal specificity of the expression of sequences included in the vector. Termination regions also facilitate effective expression at the end of transcription at appropriate points. Useful termites for practicing this invention include, but are not limited to, pinll (see An et al., Plant Cell 1 (1): 115-122 (1989)), glbl (see Genbank Access # L22345), gz (see the gzw64a terminator, Genbank Access # S78780), and the Agrobacterium bis enhancer. The termination region may be native to the nucleotide sequence of the promoter, may be native to the DNA sequence of interest, or may be derived from another source. For example, other convenient termination regions are available from the Ti plasmid of A. tumefaciens, such as the octopine synthase and nopaline synthase termination regions. See also: Guerineau et al., (1991) Mol. Gen. Genet. 262: 141-144; Proudfoot (1991) Cell 64: 671-674; Sanfacon et al., (1991) Genes Dev. 5: 141-149; Mogen et al., (1990) Plant Cell 2: 1261-1272; Munroe et al., (1990) Gene 91: 151-158; Bailas et al., 1989) Nucleic Acids Res. 17: 7891-7903; and Joshi et al., (1987) Nucleic Acid Res. 15: 9627-9639. Among the known eukaryotic promoters suitable for generalized expression are the CMV immediate early promoter, the HSV thymidine kinase promoter, the SV40 early and late promoters, the retroviral LTRs promoters, such as those of the Rous sarcoma virus.

("RSV"), metallothionein promoters, such as the mouse metallothionein I promoter and various plant promoters, such as globulin-1. When available, the native promoters of the ACC synthase genes can be used. Representatives of prokaryotic promoters include the phage lambda PL promoter, the lac trp and Tac promoters of E. coli to mention just a few of the well-known promoters. Isolated or recombinant plants, such as plant cells, that incorporate the ACC synthase nucleic acids are a feature of the invention. The transformation of plant cells and protoplasts can be carried out essentially in any of the various ways known to those skilled in the art of plant molecular biology including, but not limited to, the methods described herein. See, in general, Methods in Enzymology, Volume 153 (Recombinant DNA Part D) Wu and Gross an (eds.) 1987, Academic Press, incorporated herein by reference. As used herein, the term "transformation" means the alteration of the genotype of a host plant by the introduction of a nucleic acid sequence, eg, a "heterologous", "exogenous" or "foreign" nucleic acid sequence. . The heterologous nucleic acid sequence does not necessarily need to originate from a different source but instead, at some point, it will have been external to the cell in which it is introduced. In addition to Berger, Ausubel and Sambrook, useful general references for the cloning of plant cells, culture and regeneration include Jones (ed) (1995) Plant Gene Transfer and Expression Protocols-Methods in Molecular Biology, Volume 49 Humana Press Towata NJ; Payne and collaborators, (1992) Plant Cell and Tissue Culture in Liquid Systems John Wiley & Sons, Inc. New York, NY (Payne); and Gamborg and Phillips (eds) (1995) Plant Cell, Tissue and Organ Culture; Fundamental Methods Springer Lab Manual, Springer-Verlag (Berlin Heidelberg New York) (Gamborg). A variety of cell culture media or described in Atlas and Parks (eds) The Handbook of Microbiological Media (1993) CRC Press, Boca Raton, FL (Atlas). Additional information for the cultivation of plant cells is found in commercially available literature such as the Life Science Research Cell Culture Catalog (1998) from Sigma-Aldrich, Inc (St Louis, MO) (Sigma-LSRCCC) and for example, the Plant Culture Catalog and supplement (1997) from Sigma-Aldrich, Inc. (St Louis, MO) (Sigma-PCCS). Additional details regarding the cultivation of plant cells are found in Croy, (ed.) (1993) Plant Molecular Biology Bios Scientific Publishers, Oxford, U.K. See also the sections in the present entitled "Transformations of plants". In one embodiment of this invention, recombinant vectors including one or more of the ACC synthase nucleic acids or a subsequence thereof, for example, selected from SEQ ID NO: SEQ ID NO: 6 or SEQ ID NO: 10 , suitable for the transformation of plant cells are prepared. In another embodiment, a nucleic acid sequence encoding the desired ACC synthase RNA or protein or subsequence thereof, for example, selected from SEQ ID NOY to SEQ ID NO: 9 or SEQ ID NO: 11, is conveniently used to construct a cassette of recombinant expression that can be introduced into the desired plant. In the context of the invention, an expression cassette will typically comprise a sequence or sub-sequence of ACC synthase nucleic acid selected in an RNA configuration (eg, antisense, sense, silencing or RNA interference configuration and / or the like) operably linked to a promoter sequence and other transcriptional and transcriptional initiation regulatory sequences that are sufficient to direct transcription of the R? A configuration sequence of ACC synthase in the proposed tissues (e.g., whole plant, leaves, anthers, roots, etc.) of the transformed plant. In general, the particular promoter used in the expression cassette in plants depends on the proposed application. Any of a number of promoters may be adequate. For example, nucleic acids can be combined with constitutive, inducible, tissue-specific (tissue-preferred) promoters or other promoters for expression in plants. For example, a Strongly or weakly constitutive plant promoter that directs the expression of an RNA configuration sequence of ACC synthase in all tissues of a plant can be favorably employed. Such promoters are active under most environmental conditions and states of cell development or differentiation. Examples of constitutive promoters include the 1 'or 2' promoter of Agrobacterium tumefaciens (see, for example, O'Grady (1995) Plant Mol. Biol. 29: 99-108). Other plant promoters include the small subunit promoter of ribulose-1, 3-bisphosphate carboxylase, the phaseolin promoter, promoters of the alcohol dehydrogenase (Adh) gene (see, eg, Millar (1996) Plant Mol. Biol. 31: 897-904), sucrose synthase promoters, α-tubulin promoters, actin promoters, such as the actin Arabidopsis gene promoter (see, for example, Huang (1997) Plant Mol. Biol. 1997 33: 125-139), cab, PEPCase, R gene complex, ACTII of Arabidopsis (Huang et al.

Plant Mol. Biol. 33: 125-139 (1996)), Cat3 of Arabidopsis (Zhong et al., Mol. Gen. Biol. 251: 196-203 (1996)), the gene encoding the stearoyl acyl desaturase carrier protein of Brasica napus (Solocombe et al. (1994) Plant Physiol. 104: 1167-1176), corn GPcl (Martinez et al., (1989) J. Mol. Biol 208: 551-565), corn Gpc2 (Manjunath et al., (1997), Plant Mol. Biol. 33:97 -112) and other regions of transcription initiation of several plant genes known to those experts. See also Holtorf (1995) "Comparison of different constitutive and inducible promoters for the overexpression of transgenes in Arabidopsis thaliana", Plant Mol. Biol. 29: 637-646. The promoter sequence of the E8 gene (see, Deikman and Fischer (1988) EMBO J 7: 3315) and other genes can also be used, together with specific promoters for monocot species (for example, McElroy D. et al., (1994.) Foreign gene expression in transgenic cereals. Biotech Trends. , 12: 62-68). Other constitutive promoters include, for example, the core promoter of the Rsyn7 promoter or other constitutive promoters disclosed in WO 99/43838 and U.S. Patent No. 6,072,050; the core CaMV 35S promoter (Odell et al., (1985) Nature 313: 810-812); rice actin (McElroy et al., (1990) Plant Cell 2: 163-171); ubiquitin (Christensen et al., (1989) Plant Mol. Biol. 12: 619-632 and Christensen et al., (1992) Plant Mol. Biol. 18: 675-689); pEMU (Last et al., (1991) Theor, Appl. Genet, 81: 581-588); MAS (Velten et al., (1984) EMBO J. 3: 2723-2730); ALS promoter (U.S. Patent No. 5,659,026) and the like. Still other constitutive promoters include, for example, U.S. Patent Nos. 5,608,149; 5,608,144; 5,604,121; 5,569,597; 5,466,785; 5,399,680; 5,268,463; 5,608,142; and 6,177,611.

In addition to the promoters mentioned herein, promoters of bacterial origin operating in plants can be used in the invention. They include, for example, octopine synthase promoter, nopaline synthase promoter and other promoters derived from Ti plasmids. See, Herrera-Estrella and collaborators, (1983) Nature 303: 209. Viral promoters can also be used. Examples of viral promoters include the 35S and 19S RNA promoters of cauliflower mosaic virus (CaMV). See, Odell et al., (1985) Nature 313: 810; and Dagless (1997) Arch. Virol. 142: 183-191. Other examples of constitutive promoters of viruses that infect plants include the tobacco mosaic virus promoter; 19S and 35S promoters of cauliflower mosaic virus (CaMV) or the escrofularia mosaic virus promoter, for example, the 35S promoter of the scrofularia mosaic virus (see, for example, Maiti (1997) Transgenic Res. 6: 143 -156), etc. Alternatively, novel promoters with useful characteristics can be identified from any viral, bacterial or plant source by methods, including sequence analysis, entrapment of enhancer or promoter and the like, known in the art. Preferred promoters and enhancers of tissues (tissue-specific) can be used to direct the expression of the improved gene within a plant tissue particular. Preferred tissue tissues (tissue-specific) include, for example, those described in Yamamoto et al., (1997) Plant J. 12 (2): 255-265; Kawamata et al., (1997) Plant Cell Physiol. 38 (7): 792-803; Hansen et al., (1997) Mol. Gen Genet. 254 (3): 337-343; Russell et al., (1997) Transgenic Res. 6 (2): 157-168; Rinehart et al., (1996) Plant Physiol. 112 (3): 1331-1341; Van Camp et al., (1996) Plant Physiol. 112 (2): 525-535; Canevascini et al., (1996) Plant Physiol. 112 (2): 513-524; Yamamoto et al., (1994) Plant Cell Physiol. 35 (5): 773-778; Lam (1994) Results Probl. Cell Differ. 20: 181-196; Orozco et al., (1993) Plant Mol Biol. 23 (6): 1129-1138; Matsuoka et al. (1993) Proc Nati. Acad. Sci. USA 90 (20): 9586-9590; and Guevara-García et al., (1993) Plant J. 4 (3): 495-505. Such promoters can be modified, if necessary, for weak expression. In certain embodiments, leaf-specific promoters may be used, for example, pyruvate, orthoprosphate di-kinase (PPDK) promoter from the C4 plant (corn), cab-ml Ca + 2 promoter from corn, the gene promoter related to myb from Arabidopsis thaliana (promoter Atmyb5), the ribulose bisphosphate carboxylase promoters (RBCS) (for example, tomato genes RBCS1, RBCS2, RBCS3A, which are expressed in leaves and seedlings cultured with light, whereas RBCS1 and RBCS2 are expressed in developing tomato fruits and / or the ribulose bisphosphate carboxylase promoter which is expressed almost exclusively in mesophilic cells in leaves and leaf covers at high levels, etc.) and the similar ones. See, for example, Matsuoka et al., (1993) Tissue-specific light-regulated expression directed by the promoter of a C4 gene, maize pyruvate, orthophosphate dikinase, in a C3 plant, rice, PNAS USA 90 (20): 9586- 90; (2000) Plant Cell Physiol. 41 (1): 42-48; (2001) Plant Mol. Biol. 45 (1): 1-15; Shiina, T. et al., (1997) Identification of Promoter Elements involved in the cytosolic Ca + 2 mediated photoregulation of maize cab-ml expression, Plant Physiol. 115: 477-483; Casal (1998) Plant Physiol. 116: 1533-1538; Li (1996) FEBS Lett. 379: 117-121; Busk (1997) Plant J. 11: 1285-1295; and Meier (1997) FEBS Lett. 415: 91-95; and Matsuoka (1994) Plant J. 6: 311-319. Other leaf-specific promoters include, for example, Yamamoto et al., (1997) Plant J. 12 (2): 255-265; Kwon et al., (1994) Plant Physiol. 105: 357-67; Yamamoto et al., (1994) Plant Cell Physiol. 35 (5): 773-778; Gotor et al., (1993) Plant J. 3: 509-18; Orozco et al., (1993) Plant Mol. Biol. 23 (6): 1129-1138; and Matsuoka et al., (1993) Proc. Nati Acad. Sci. USA 90 (20): 9586-9590. In certain modalities, the specific promoters of senescence can be used (for example, an active tomato promoter during fruit ripening, senescence and abscission of leaves, a maize promoter of the gene encoding a cysteine protease and the like). See, for example, Blume (1997) Plant J. 12: 731-746; Griffiths et al., (1997) Sequencing, expression pattern and RFLP mapping of a senescence-enhanced cDNA from Zea Mays wi th high homology to oryzain gamma and aleurain, Plant Mol. Biol. 34 (5): 815-21; The seel gene partially from Zea is for cysteine protease, promoter region and 5 'coding region, Genbank AJ494982; Kleber-Janke, T. and Krupinska, K. (1997) Isolation of cDNA clones for genes showing enhanced expression in barley leaves during dark-induced senescence as well as during senescence underfield conditions, Planta 203 (3): 332-40; and Lee, RH et al., (2001) Leaf senescence in rice plants: cloning and characterization of senescence up ~ regulated genes, J. Exp. Bot. 52 (358): 1117-21. In other modalities, anther-specific promoters may be used. Such promoters are known in the art or can be discovered by known techniques; see, for example, Bhalla and Singh (1999) Molecular control of male fertility in Brassica Proc. 10th Annual Rapeverd Congress, Canberra. Australia; van Tunen and collaborators, (1990) Pollen ~ and anther-specific chi promoters from petunia; tandem promoter regulation of the chiA gene. Plant Cell 2: 393-40; Jeon et al., (1999) Isolation and characterization of an anther specific gene, RA8, from rice (Oryza sativa L). Plant Molecular Biology 39: 35-44; and Twell et al., (1993) Activation and developmental regulation of an Arabidopsis anther-specific promoter in microspores and pollen of Nicotiana tabacum. Sex Plant Reprod. 6: 217-224. Preferred root promoters are known and can be selected from the many available from the literature or de novo isolates of several compatible species. See, for example, Hire et al., (1992) Mol. Biol. 20 (2): 207-218 (glutamine synthetase gene specific to soybean root); Keller and Baumgartner (1991) Plant Cell 3 (10): 1051-1061 (root specific control element in the GRP 1.8 gene of green bean); Sanger et al., (1990) Mol. Biol. 14 (3): 433-443 (root specific promoter of the mannopin synthase (MAS) gene of Agrobacterium tumefaciens); and Miao et al., (1991) Plant Cell 3 (l): ll-22 (full-length cDNA clone encoding cytosolic glutamine synthetase (G), which is expressed in roots and nodules of soybean root). See also Bogusz et al., (1990) Plant Cell 2 (7): 633-641, where two root-specific promoters isolated from hemoglobin genes from the non-nitrogen-fixing Parasponia andersonii and the non-nitrogen-fixing non-ligneous vegetable related Trema Tomentosa are described. The promoters of these genes were linked to a reporter gene of ß-glucuronidase e were introduced in both the non-legume Nicotiana tabacum and the legume Lotus corniculatus, and in both cases the activity of the specific root promoter was preserved. Leach and Aoyagi (1991) describe their analysis of the promoters of the rolC induction genes and rolD highly expressed Agrobacterium rhizogenes (see Plant Science (Limerick) 79 (1): 69-76). They concluded that the enhancer and the preferred tissue DNA determinants are dissociated in those promoters. Teeri et al. (1989) used gene fusion to lacZ to show that the Agrobacterium T-DNA gene encoding octopine synthase is especially active in the epidermis of the root tip and that the TR2'g gene is specific to the root in the plant intact and stimulated by leaf tissue injury (see, for example, EMBO J. 8 (2): 343-350). The Gen TRl ', fused to nptll (neomycin phosphotransferase II) showed similar characteristics. Preferred promoters of additional roots include the VfENOD-GRP3 gene promoter (Kuster et al., (1995) Plant Mol. Biol. 29 (4): 759-772); and rolB promoter (Capana et al., (1994) Plant Mol. Biol. 25 (4): 681-691 See also, U.S. Patent Nos. 5,837,876, 5,750,386, 5,633,363, 5,459,252, 5,401,836, 5,110,732, and 5,023,179. "seed preferences" include both "seed-specific" promoters those active promoters during seed development such as promoters of seed storage proteins) as well as "seed germination" promoters (those active promoters during germination of the seed). See, for example, Thompson et al., (1989) BioEssays 10: 108, incorporated herein by reference. Such preferred seed promoters include, but are not limited to, Ciml (induced cytokinin message); cZ19Bl (19 kDa corn zein); milpas (myo-inositol-1-phosphate synthase); mZE40-2, also known as Zm-40 (U.S. Patent No. 6,403,862); Nuclc (U.S. Patent No. 6,407,315); and celA (cellulose synthase) (see WO 00/11177). The gamma-zein is a specific endosperm promoter. Glob-1 is a specific promoter of the embryo. For dicotyledons, seed-specific promoters include, but are not limited to bean β-phaseolin, napkin, β-conglycinin, soybean lectin, cruciferin and the like. For monocotyledons, seed-specific promoters include, but are not limited to, the 15 kDa corn zein promoter, a 22 kDa zein promoter, a 27 kDa zein promoter, a G-zein promoter, a promoter ? -27 kD zein (such as the gzw64A promoter, see Genbank Access # S78780), a waxy promoter, a shunken 1 promoter, a shunken 2 promoter, a globulin 1 promoter (see Genbank Access # L22344), an lt promoter? 2 (Kalla and collaborators, Plant Journal 6: 849-860 (1994); U.S. Patent No. 5,525,716), ciml promoter (see U.S. Patent No. 6,225, 529) endl and end2 promoters of corn (See U.S. Patent No. 6,528,704 and application 10 / 310,191, filed December 4, 2002); promoter nuci (US Patent No. 6,407,315); promoter Zm40 (U.S. Patent No. 6,403,862); eepl and eep2; lecl (US patent application No. 09 / 718,754); thioredoxin H promoter (US provisional patent application 60 / 514,123); mlipl5 promoter (U.S. Patent No. 6,479,734); PCNA2 promoter; and the shrunken-2 promoter. (Shaw et al., Plant Phys 98: 1214-1216, 1992; Zhong Chen et al., PNAS USA 100: 3525-3530, 2003). However, other promoters useful in the practice of the invention are known to those skilled in the art. such as the promoter nucelain (See C. Linnestad et al., Nucellain, A Barley Homolog of the Dicote Vacuolar-Processing Protease is Localized in Nucellar Cell Walls, Plant Physiol. 118: 1169-80 (1998), promoter knl (See S. Hake and N. Ori, The Role of Knottedl in Meristem Functions, B8: INTERACTIONS AND INTERSECTIONS IN PLANT PATHWAYS, COEUR D 'WOOD, IDAHO, KEYSTONE SY POSIA, February 8-14, 1999, to 27.) and promoter F3.7 (Baszczynski et al., Maydica 42: 189-201 (1997)), etc. In certain modalities, the promoters that act spatially such as glbl, a promoter is preferred of embryo; or zein range, a preferred endosperm promoter; or an active promoter in the surrounding region of the embryo (see US patent application No. 10 / 786,679, filed on February 25, 2004) or BETL1 (See G. Hueros et al., Plant Physiology 121: 1143-1152 (1999 ) and Plant Cell 7: 747-57 (June 1995)), are useful, including preferentially active promoters in female reproductive tissues, and those active in meristematic tissues, particularly female meristematic reproductive tissues. See also, WO 00/12733, where the seed preferred promoters of the endl and end2 genes are disclosed. A tissue-specific promoter can induce the expression of operably linked sequences in tissues other than the target tissue. Thus, as used herein, a tissue-specific promoter is one that induces expression preferentially in the target tissue, but may lead to some expression in other tissues as well. The use of temporarily acting promoters is also contemplated by this invention. For example, promoters that act 0-25 days after pollination (DAP), 4-21, 4-12 or 8-12 of DAP can be selected, for example, promoters such as ciml and ltp2. Promoters that act from -14 to 0 days after pollination can also be used, such as SAG12 (See WO 96/29858, Richard M. Amasino, published on October 3, 1996) and ZAG1 or ZAG2 (See RJ Schmidt et al., Identification and Molecular Characterization of ZAGI, the Maize Homolog of the Arabidopsis Floral Homeotic Gene AGAMOUS, Plant-Cell 5 (7): 729-37 (July 1993)). Other useful promoters include zag2.1 of corn, Zap (also known as ZmMADS, U.S. Patent Application No. 10 / 387,937, WO 03/078590); and the corn tbl promoter (see also Hubbarda et al., Genetics 162: 1927-1935, 2002). Where overexpression of a nucleic acid of ACC synthase RNA configuration is detrimental to the plant, one skilled in the art will recognize that weak constitutive promoters can be used for low levels of expression (or, in certain embodiments, inducible or specific promoters may be used). of tissue). In those cases, where high levels of expression are not harmful to the plant, a strong promoter can be used, for example, a T-RNA, or another pol III promoter, or strong pol II promoter (e.g., the virus promoter). mosaic cauliflower, 35S CaMV promoter). Where low expression level is desired, weak promoters will be used. Generally, by "weak promoter" a promoter is proposed which induces the expression of a coding sequence at a low level. For low level levels of approximately 1/1000 transcripts are proposed to approximately 1 / 100,000 transcripts to approximately 1 / 500,000 transcripts. Alternatively, it is recognized that weak promoters also comprise promoters that induce expression in only a few cells and not in others to give a low overall level of expression. Where a promoter induces expression at unacceptably high levels, portions of the promoter sequence can be deleted or modified to decrease expression levels. Such weak constitutive promoters include, for example, the core promoter of the Rsyn7 promoter (WO 99/43838 and US Patent No. 6,072,050), the 35S promoter of core CaMV and the like. In certain embodiments of the invention, an inducible promoter can be used. For example, a plant promoter may be under environmental control. Such promoters are referred to as "inducible" promoters. Examples of environmental conditions that can alter transcription by inducible promoters include pathogen attack, anaerobic conditions, elevated temperature and the presence of light. For example, the invention incorporates the drought-inducible promoter of corn (Busk (1997) Piant J. 11: 1285-1295); the cold inducible promoter, drought, high salt concentration of the potato (Kirch (1997) Plant Mol. Biol. 33: 897-909) and the like. Pathogen-inducible promoters include those of proteins related to pathogenesis (PR proteins), which are induced after infection by a pathogen; for example, PR proteins, S7? R proteins, beta-1, 3-glucanase, chitinase, etc. See, for example, Redolfi et al. (1983) Neth. J. Plant Pathol. 89: 245-254; Uknes et al., (1992) Plant Cell 4: 645-656; and Van Loon (1985) Plant Mol. Virol. 4: 111-116. See also the application entitled "Inducible Maize Promoters", US patent application serial number 09 / 257,583, filed on February 25, 1999. Of interest are the promoters that are locally expressed at or near the site of the pathogen's infection. Marineau et al., (1987) Plant Mol. Biol. 9: 335-342; Matton et al. (1989) Molecular Plant-Microbe Interactions 2: 325-331; So sisch et al., (1986) Proc. Nati Acad. Sci. 83: 2427-2430; Somsisch et al. (1988) Mol. Gen. Genet. 2: 93-98; and Yang (1996) Proc. Nati Acad. Sci. 93: 14972-14977. See also, Chen et al., (1996) Plant J. 10: 955-966; Zhang et al., (1994) Proc. Nati Acad. Sci. 91: 2507-2511; Warner et al., (1993) Plant J. 3: 191-201; Siebertz et al., (1989) Plant Cell 1: 961-968; U.S. Patent No. 5,750,386 (inducible by nematode) and the references cited therein. Of particular interest is the inducible promoter for the corn PRmS gene, whose expression it is induced by the pathogen Fusarium moniliforme (see, for example, Cordero et al., (1992) Physiol., Mol. Plant Path, 41: 189-200). Additionally, as pathogens find entry into plants through injury or insect damage, a promoter inducible by injury can be used in the constructions of the invention. Such injury-inducible promoters include the potato proteinase inhibitor (pin II) (Ryan (1990) Ann. Rev. Phytopath., 28: 425-449; Duan et al., (1996) Nature Biotechnology 14: 494-498); wunl and wun2, U.S. Patent No. 5,428, 148; winl and win2 (Stanford et al., (1989) Mol. Gen. Genet. 215: 200-208); systemin (McGurl et al., (1992) Science 225: 1570-1573); W1P1 (Rohmeier et al., (1993) Plant Mol. Biol. 22: 783-792; Eckelkamp et al. (1993) FEBS Letters 323: 73-76); MPI gen (Corderok et al., (1994) Plant J. 6 (2): 141-150); and the similar ones. Alternatively, plant promoters that are inducible upon exposure to plant hormones, such as auxins, are used to express the polynucleotides of the invention. For example, the invention may use the El promoter subsequence of auxin response elements (AuxREs) from soybean (Glycine Max L ..) (Liu (1997) Planta Physiol. 115: 397-407); the GST6 promoter of Arabidopsis responsive to auxin (also responsive to salicylic acid and hydrogen peroxide) (Chen (1996) Plant J. 10: 955-966); the auxin-inducible parC promoter of tobacco; a plant biotin response element (Streit (1997) Mol Plant Microbe Interact 10: 933-937); and the abscisic acid responsive promoter of stress hormone (Sheen (1996) Science 274: 1900-1902). Plant promoters that are inducible upon exposure to chemical reagents that can be applied to the plant, such as herbicides or antibiotics, are also used to express the polynucleotides of the invention. Depending on the objective, the promoter can be a promoter inducible by chemical substance, where the application of the chemical induces the expression of the gene, or a repressible promoter by chemical substance, where the application of the chemical represses the expression of the gene. For example, the corn In2-2 promoter, activated by moderates of benzenesulfonamide herbicide, can be used (De Veylder (1997) Plant Cell Physiol. 38: 568-577); the application of different herbicide moderates induces different gene expression patterns, including root expression, hydathodes, and apical shoot meristem. A coding sequence of ACC synthase or RNA configuration may also be under the control of, for example, tetracycline-inducible and repressible tetracycline promoters. (see, for example, Gatz et al., (1991) Mol. Gen. Genet., 227: 229-237, U.S. Patent Nos. 5,814,618 and 5,789,156, and, Masgrau (1997) Plant J. 11: 465-473 (description of transgenic tobacco plants containing the arginine decarboxylase gene of Avena sativa L (oats) with a tetracycline-inducible promoter), or, a responsive element to salicylic acid (Stange (1997) Plant J. 11: 1315-1324.) Other inducible promoters by chemical substances are known in the art and include, but are not limited to, the corn GST promoter, which is activated by hydrophobic electrophilic compounds that are used as pre-emergent herbicides, and the tobacco PR promoter, which is activated by Salicylic acid Other promoters regulated by chemical substances of interest include the steroid responsive promoters (see, for example, the glucocorticoid-inducible promoter Schena et al., (1991) Proc. Nati. Acad. Sci. USA 88: 10421-10425 and Mc Nellis et al., (1998) Plant J. 14 (2) .- 247-257). Endogenous promoters of genes related to herbicide tolerance and related phenotypes are also useful for inducing the expression of nucleic acids from the ACC synthase RNA configuration, eg, P450 Monooxygenases P450, glutathione S-transferase, homoglutathione S-transferases , glyphosate oxidases and 5- enolpiruvilshiquiimato-2-phosphate synthases. For example, a plant promoter linked to a polynucleotide of the invention may be useful when it is desired to flip the expression in the presence of a particular condition, eg, drought conditions, short growth conditions, density, etc. Tissue-specific promoters can also be used to direct the expression of the polynucleotides of the invention, including the nucleic acids of ACC synthase RNA configuration, such that a polynucleotide of the invention is expressed only in certain tissues or stages of development, for example, leaves, anthers, roots, shoots, etc. Tissue-specific expression may be advantageous, for example, when expression of the polynucleotide in certain tissues is desirable while expression in other tissues is undesirable. Tissue-specific promoters are the transcriptional control elements that are only active in particular cells or tissues at specific times during the development of the plant, such as in vegetative tissues or reproductive tissues. Examples of tissue-specific promoters under development control include promoters that initiate transcription only (or principally only) in certain tissues, such as vegetative tissues, eg, roots or leaves, or tissues reproducers, such as the fruit, ovules, seeds, pollen, pistols, flowers or any embryonic tissue. Reproductive-specific promoters may be, for example, anther-specific, ovum-specific, embryo-specific, endosperm-specific, integument-specific, seed-specific and seed-coat-specific, pollen-specific, petal-specific, Separate it, or some combination of them. It will be understood that numerous promoters not mentioned are suitable for use in this aspect of the invention, are well known and can easily be employed by those skilled in the manner illustrated by the discussion and examples herein. For example, this invention contemplates the use, when appropriate, of native ACC synthase promoters to induce the expression of the enzyme (or sequences or subsequences of ACC synthase polynucleotide) in a recombinant environment. In the preparation of expression vectors of the invention, sequences other than those associated with the endogenous ACC synthase gene, mRNA or polypeptide sequence, or subsequence thereof, can optionally be used. For example, other regulatory elements such as introns, leader sequences, polyadenylation regions, signal / location peptides, etc. They can also be included.

The vector comprising a polynucleotide of the invention may also include a marker gene that confers a selectable phenotype on plant cells. For example, the marker can encode the biocidal tolerance, particularly antibiotic tolerance, such as tolerance to kanamycin, G418, bleomycin, hygromycin or herbicide tolerance, such as tolerance to chlorosulfuron, or fofinothricin. Reporter genes that are used to monitor gene expression and protein localization via visualizable reaction products (eg beta-glucuronidase, beta-galactosidase and chloramphenicol acetyltransferase) or by direct visualization of the gene product same (eg, green fluorescent protein, GFP; Sheen et al., (1995) The Plant Journal 8: 777) can be used to, for example, monitor the expression of transient gene in plant cells. Vectors for propagation and expression will generally include selectable markers. Such markers may also be suitable for amplification or the vectors may contain additional markers for this purpose. In this regard, the expression vectors preferably contain one or more selectable marker genes to provide a phenotypic attribute for the selection of transformed host cells. The Preferred markers include dihydrofolate reductase or neomycin resistance for the culture of eukaryotic cells, and resistance genes to tetracycline or ampicillin for the culture of E. coli and other prokaryotes. The kanamycin and herbicide resistance genes (PAT and BAR) are generally useful in plant systems. Selectable marker genes, in physical proximity to the introduced DNA segment, are used to allow transformed cells to be recovered either by positive genetic selection or classification. Selectable marker genes also allow the selection pressure to be maintained in a population of transgenic plants, to ensure that the introduced DNA segment, and its promoters and control enhancers, are retained by the transgenic plant. Many of the positive selectable marker genes commonly used for the transformation of plants have been isolated from bacteria and encode enzymes that detoxify a selective chemical agent that can be an antibiotic or a herbicide. Other positive selection marker genes encode an altered target that is insensitive to the inhibitor. An example of a selection marker gene for plant transformation is the BAR or PAT gene, which is used with the bialaphos selection agent (Spencer et al.

J. Theor. Appl'd Genetics 79: 625-631 (1990)). Other Gen The useful selection marker is the neomycin phosphotransferase II [nptll] gene, isolated from Tn5, which confers resistance to kanamycin when placed under the control of regulatory signals from the plant (Fraley et al., Proc. Nat'l Acad. Sci. USA) 80: 4803 (1983)). The hygromycin phosphotransferase gene, which confers resistance to the hygromycin antibiotic, is an additional example of a useful selectable marker (Vanden Elzen et al., Plant Mol. Biol. 5: 299 (1985)). Additional positive selectable marker genes of bacterial origin that confer resistance to antibiotics include gentamicin acetyltransferase, streptomycin phosphotransferase, aminoglycoside-3'-adenyltransferase and the determinant of gleoraicin resistance (Hayford et al., Plant Physiol. 86: 1216 (1988); Jones et al., Mol. Gen.

Genet 210: 86 (1987); Svab et al., Plant Mol. Biol. 14: 197 (1990); Hille et al., Plant Mol. Biol. 7: 171 (1986)). Other selectable positive marker genes for plant transformation are not of bacterial origin. These genes include mouse dihydrofolate reductase, plant 5-enolvilshiquimato-3-phosphate synthase and plant acetolactate synthase (Eichholtz et al., Somatic Cell Mol. Genet., 13:67 (1987); Shah et al.

Science 233: 478 (1986); Charest et al., Plant Cell Rep. 8: 643 (1990)). Other examples of suitable selectable marker genes include, but are not limited to: genes encoding chloramphenicol resistance, Herrera Estrella et al. (1983) EMBO J. 2: 987-992; metrotrexate, Herrera Estrella et al. (1983) Nature 303: 209-213; Meijer et al. (1991) Plant Mol. Biol. 16: 807-820; hygromycin, Waldron et al. (1985) Plant Mol. Biol. 5: 103-108; Zhijian et al. (1995) Plant Science 108: 219-227; Streptomycin, Jones et al. (1987) Mol. Gen Genet 210: 86-91; Spectinomycin, Bretagne-Sagnard et al. (1996) Transgenic Res. 5: 131-137; bleomycin, Hille et al. (1990) Plant Mol. Biol. 7: 171-176; sulfonamide, Guerineau et al. (1990) Plant Mol. Biol. 15: 127-136; Bromoxynil, Stalker et al (1988) Science 242: 419-423; glyphosate, Shaw et al. (1986) Science 233: 478-481; phosphinothricin, DeBlock et al. (1987) EMBO J. 6: 2513-2518. Another class of marker genes useful for transforming plants with the DNA sequence requires the classification of presumably transformed plant cells rather than the direct genetic selection of transformed cells for resistance to a toxic substance such as an antibiotic. These genes are particularly useful for quantifying or visualizing the spatial pattern of the expression of the DNA sequence in specific tissues and are frequently referred to as reporter genes because they can be fused to a gene or regulatory sequence of the gene for the investigation of gene expression. Genes commonly used to classify presumably transformed cells include β-glucuronidase (GUS), β-galactosidase, luciferase and chloramphenicol acetyltransferase (Jefferson, Plant Mol. Biol. Rep. 5: 387 (1987); Teeri et al., EMBO J. 8 : 343 (1989), Koncz et al, Proc. Nat and Acad. Sci. (USA) 84: 131 (1987), De Block et al., EMBO J. 3: 1681 (1984)). Examples of other suitable reporter genes known in the art can be found in, for example: Jefferson et al. (1991) in Plant Molecular Biology Manual, ed. Gelvin et al. (Kluwer Academic Publishers), pp. 1-33; DeWet et al. (1987) Mol. Cell. Biol. 7: 725-737; Goff et al. (1990) EMBO J. 9: 2517-2522; Kain et al. (1995) BioTecZmiques 19: 650-655; and Chiu et al. (1996) Current Biology 6: 325-330. Another method for the identification of relatively rare transformation evidences has been the use of a gene encoding a dominant constitutive regulator of the anthocyanin pigmentation pathway of Zea mays (Ludwig et al. Science 247: 449 (1990)).

The appropriate DNA sequence can be inserted into the vector by any of the well-known and routine techniques. In general, a DNA sequence for expression is linked to an expression vector by cleaving the DNA sequence and the expression vector with one or more restriction endonucleases and then by joining the restriction fragments together using T4 DNA ligase. The sequence can be inserted in a forward or backward orientation. The procedures for restriction and ligation that can be used for this purpose are well known and routine for those skilled in the art. Suitable procedures in this regard, and for the construction of expression vectors using alternative techniques, which are also well known and routine for those skilled in the art, are set forth in great detail in Sambrook et al. MOLECULAR CLONING, A LABORATORY MANUAL, 2nd Ed .; Cold Spring Harbor Laboratory Press, Cold Spring Harbor, Nes York (1989). A polynucleotide of the invention, optionally encoding the heterologous structural sequence of a polypeptide of the invention, would generally be inserted into the vector using standard techniques so that it is operably linked to the promoter for expression.

(Operably, linked as used herein, includes reference to a functional link between a promoter and a second sequence, wherein the promoter sequence initiates and mediates the transcription of the DNA corresponding to the second sequence. Generally, operably linked means that the nucleic acid sequences that are linked are contiguous and, where necessary, they bind two protein coding regions, contiguous and in the same reading structure). When the polynucleotide is proposed for the expression of a polypeptide, the polynucleotide will be positioned so that the transcription initiation site is appropriately located 5 'to a ribosome binding site. The ribosome binding site will be 5 'to the AUG that initiates the translation of the polypeptide to be expressed. Generally, there will be no other open reading structures that start with an initiation codon, usually AUG, and lie between the ribosome binding site, and the initiation codon. Also, generally, there will be a stop codon of translation at the end. of the polypeptide and there will be a polyadenylation signal in the constructions for use in eukaryotic hosts. Properly arranged transcription termination signals in the 3 'system of the transcribed region can also be included in the polynucleotide construct. For nucleic acid constructs designed to characterize a polypeptide, the expression cassettes may additionally contain 5 'leader sequences. Such Guide sequences can act to increase translation.

Translation guides are known in the art and include: picornavirus guides, for example: EMCV guide (5 'non-coding region of Encephalomyocarditis), Elroy-Stein et al. (1989) Proc. Nat. Acad. Sci. USA 86: 6126-6130; potivirus guides, for example, TEV (Tobacco Etch Virus) guide, Allison et al. (1986); MDMV guide (Dwarf Corn Mosaic Virus), Virology 154: 9-20; human immunoglobulin heavy chain binding protein (BiP), Macejak et al. (1991) Nature 353: 90-94; untranslated guide of mRNA of alfalfa mosaic virus coating protein (AMV RNA 4), Jobling et al. (1987) Nature 325: 622-625); tobacco mosaic virus (TMV) guide, Gallie et al. (1989) Molecular Biology of RNA, pages 237-256; and guidance of corn chlorotic spotted virus (MCMV) Lommel et al. (1991) Virology 81: 382-385. See also Della-Cioppa et al. (1987) Plant Physiology 84: 965-968. The cassette may also contain sequences that increase the translation and / or stability of mRNAs such as introns. The expression cassette also typically includes, at the 3 'terminus of the isolated nucleotide sequence of interest, a translation termination region, eg, a functional one in plants.

In those cases where it is desirable to have the expressed product of the isolated nucleotide sequence directed to a particular organelle, particularly the plastid, amiloplast or endoplasmic reticulum, or secreted on the surface of the cell or extracellularly, the expression cassette may further comprise a coding sequence for a transit peptide. Such transit peptides are well known in the art and include, but are not limited to: the transit peptide for the acyl carrier protein, the small subunit of RUBISCO, plant EPSP synthase and the like. In the preparation of the expression cassette, the various DNA fragments can be manipulated, to provide the DNA sequences in the proper orientation and, as appropriate, the appropriate reading structure. Towards this end, adapters or linkers can be used to join DNA fragments or other manipulations can be involved to provide convenient restriction sites, removal of superfluous DNA, removal of restriction sites or the like. For this purpose, it may be involved in in vitro mutagenesis, primer repair, restriction digestions, annealing and substitutions such as transitions and transversions.

As mentioned herein, the present invention provides vectors capable of expressing genes of interest under the control of regulatory elements. In general, the vectors can be functional in plant cells. Sometimes, it may be preferable to have vectors that are functional in other host cells, for example, in E. coli (e.g., for the production of protein to highlight antibodies, DNA sequence analysis, construction of inserts, or obtain amounts of nucleic acids) . Vectors and methods for cloning and expression in E. coli are discussed in Sambrook et al. (Supra). The transformation vector, comprising the promoter of the present invention operably linked to an isolated nucleotide sequence in an expression cassette, may also contain at least one additional nucleotide sequence for a gene that is cotransformed in the organism. Alternatively, the additional sequence (s) may be provided in another transformation vector. The vector containing the appropriate DNA sequence as described elsewhere herein, as well as an appropriate promoter, and other appropriate control sequences, can be introduced into an appropriate host using a variety of well-known techniques suitable for the expression therein of a desired RNA and / or polypeptide. The present invention also relates to host cells that contain the constructions described in the above. The host cell can be a higher eukaryotic cell such as the plant cell, a lower eukaryotic cell, such as a yeast cell, or the host cell can be a prokaryotic cell, such as a bacterial cell. The introduction of the construction into the host cell can be effected by calcium phosphate transfection, DEAE-dextran-mediated transfection, microinjection, cationic lipid-mediated transfection, electroporation, transduction, scratch loading, ballistic introduction, infection or other methods . Such methods are described in many standard laboratory manuals, such as Davis et al BASIC METHODS IN MOLECULAR BIOLOGY, (1986) and Sambrook et al., MOLECULAR CLONING: A LABORATORY MANUAL, 2nd Ed., Cold Spring Harbor Laboratory Press, Cold Spring Harbor , NY (1989). Representative examples of suitable hosts include bacterial cells, such as streptococcal cells, staphylococci, E. coli, streptomyces and Salmonella typhimurium; fungal cells, such as yeast cells and Aspergillus cells; insect cells such as Drosophila S2 and Spodoptera Sf9 cells; cells of animal such as CHO cells, COS and Bowes melanoma; and plant cells. Plant cells can be derived from a wide range of plant types, particularly monocotyledons such as Gramineae species including Sorghum bicolor and Zea mays, as well as dicotyledons such as soybean (Glycine max) and canola (Brassica napus, Brassica rapa ssp.). Preferably the plants include corn, soybean, sunflower, safflower, canola, wheat, barley, rye, alfalfa, rice, oats, turf grass and sorghum; however, the isolated nucleic acid and the proteins of the present invention can be used in species of the genera: Ananas, Antirrhinum, Arabidopsis, Arachis, Asparagus, Atropa, Oats, Brassica, Bromus, Browaalia, Camellia, Capsicum, Ciahorium, Ci trus, Cocos, Cofea, Cucumis, Cucurbi ta, Datura, Daucus, Digi talis, Ficus, Fragaria, Geranium, Glycine, Gossypium, Helianthus, Heterocallis , Hordeun, Hyoscyamus, Ipomoea, Juglans, Lactuca, Linum, Lolium, Lotus, Lycopersicon, Majorana, Mangifera, Manihot, Medicago, Musa, Nemesis, Nicotiana, Olea, Onobrychis, Oryza, Panieum, Pelargoniun, Pennisetum, Persea, Petunia, Phaseolus , Pisum, Psidium, Ranunculus, Raphanus, Rose, Salpiglossis, Sécale, Senecio, Solanum, Sinapis, Sorghum, Theobroma, Triticum, Trifolium, Trigonella, Vigna, Vitis and Zea, among many other examples (for example, other genera mentioned herein.) The promoter regions of the invention can be isolated from any plant, including, but not limited to, maize (corn, Zea mays), cañola (Brassica napus, Brassica rapa ssp.), Alfalfa (Medicago sativa), rice (Oryza sativa). ), rye (Sécale cereale), sorghum (Sorghum bicolor, Sorghum vulgare), sunflower (Helianthus annuus), wheat (Tri ticuln aestivum), soybean (Glycine max), tobacco (Nicotiana tabacum), potato (Solanum tuberosum), walnuts ( Arachis hypogaea), cotton (Gossypium hirsutum), sweet potato (Ipomoea battus), cassava (Manihot esculenta), coffee (Cofea spp.), Coco (Cocos nucifera), pineapple (Ananas comosus), citrus trees (Citrus spp.), Cacao (Theobroma cacao), te (Camellia sinensis), banana (Musa spp.), avocado (Persea americana), fig (Ficus casica), guava (Psidium guajava), mango (Mangifera indica), olive (Olea europaea), oats, barley, vegetables, plants ornamental, and coniferous. Preferably, the plants include corn, soybean, sunflower, safflower, canola, wheat, barley, rye, alfalfa, rice, oats, turf grass and sorghum. Hosts for a wide variety of expression constructs are well known, and those skilled in the art will be enabled by the present disclosure to readily select a host to express a polypeptide in accordance with this aspect of the present invention.

The designed host cells can be cultured in conventional nutrient media, which can be modified as is appropriate for, inter alia, activating promoters, selecting transformants or amplifying genes. The culture conditions, such as temperature, pH and the like, previously used with the host cell selected for expression will generally be suitable for the expression of nucleic acid and / or polypeptide of the present invention, as will be apparent to those skilled in the art. technique. Mature proteins can be expressed in mammalian cells, yeast cells, bacteria or other cells under the control of appropriate promoters. Cell-free translation systems can also be employed to produce such proteins using RNAs derived from the DNA constructs of the present invention. After transformation of a suitable host strain and growth of the host strain to an appropriate cell density, where the selected promoter is inducible, it is induced by appropriate means ( example, shift of exposure temperature to chemical inducer) and the cells are cultured for an additional period. The cells are then typically harvested by centrifugation, broken by physical or chemical means, and the resulting crude extract retained for further purification. The microbial cells used in the expression of proteins can be broken by any convenient method, including recycling of freeze-thaw, sonication, mechanical breakage or the use of cell lysate agents; such methods are well known to those skilled in the art. METHODS FOR INHIBITING ETHYLENE PRODUCTION The invention also provides methods for inhibiting the production of ethylene in a plant (and plants produced by such methods). For example, a method for inhibiting ethylene production comprises inactivating one or more ACC synthase genes in the plant, wherein the one or more ACC synthase genes encode one or more ACC synthases. Typically, at least one or more ACC synthases comprise, for example, at least about 70%, at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 99%, about 99.5% or more identity to SEQ ID NOY (pACS2), SEQ ID NO: 8 (pACS6), SEQ ID NO: 9 (pAC7) or SEQ ID NO: 11 (pCCRA178R). Antisense, Sense, Mute or RNA Interference Configurations The one or more ACC synthase genes can be inactivated by introducing and expressing transgenic sequences, for example antisense or sense configurations, or RNA silencing and interference configurations, etc., encoding one or more ACC synthases, or a subsequence of them, and a promoter, to thereby inactivate the one or more ACC synthase genes compared to a corresponding control plant (for example, its non-transgenic origin or a non-transgenic plant of the same species). See also the section entitled "Polynucleotides of the Invention". The at least one polynucleotide sequence can be introduced by techniques including, but not limited to, for example, electroporation, microprojectile bombardment, Agrobacterium-mediated transfer and other available methods. See also the section entitled "Transformation of plants" in the present. In certain aspects of the invention, the polynucleotide is linked to the promoter in a sense orientation or an antisense reorientation or is configured for RNA silencing or interference. In certain situations it may be preferable to mute or down-regulate certain genes, such as the ACC synthase genes. The relevant literature describing the application of silencing of the gene dependent on homology includes: Jorgensen, Trends Biotechnol. 8 (12): 340- 344 (1990); Flavell, Proc. Nat'l. Acad. Sci. (USA) 91: 3490-3496 (1994); Finnegan et al., Bio / Technology 12: 883-888 (1994); Neuhuber et al., Mol. Gen. Genet. 244: 230-241 (1994); Flavell et al. (1994) Proc. Nati Acad. Sci. USA 91: 3490-3496; Jorgensen et al. (1996) Plant Mol. Biol. 31: 957-973; Johansen and Carrington (2001) Plant Physiol. 126: 930-938; Broin et al. (2002) Plant Cell 14: 1417-1432; Stoutjesdijk et al. (2002) Plant Physiol. 129: 1723-1731; Yu et al. (2003) Phytochemistry 63: 753-763; and U.S. Patent Nos. 5,034,323, 5,283,184, and 5,942,657. Alternatively, another method for silencing the gene can be with the use of antisense technology (Rothstein et al., In Plant Mol. Cell, Biol. 6: 221-246 (1989); Liu et al. (2002) Plant Physiol. 1732-1743 U.S. Patent Nos. 5,759,829 and 5,942,657 The use of antisense nucleic acids is well known in the art.An antisense nucleic acid has a region of complementarity in a target nucleic acid, for example, an ACC synthase gene, an mRNA or cDNA The antisense nucleic acid can be RNA, DNA, a PNA or any other appropriate molecule A duplex can be formed between the antisense sequences and their complementary sense sequence, resulting in inactivation of the gene Antisense nucleic acid can inhibit expression of the gene when forming a duplex with an RNA transcribed from the gene, forming a triplex with the D? A duplex, etc. An antisense nucleic acid can be produced, for example, by an ACC synthase gene by a number of well-established techniques (e.g., chemical synthesis of an antisense R? A or an oligonucleotide (optionally including modified nucleotides and / or linkages that increase the resistance to degradation or improve cellular uptake) or in vitro transcription). Antisense nucleic acids and their use are described, for example, in USP 6,242,258 by Haselton and Alexander (June 5, 2001) entitled "Methods for the selective regulation of DNA and RNA transcription and translation by photoactivation"; USP 6,500,615; USP 6,498,035; USP 6,395,544; USP 5,563,050; E. Schuch et al. (1991) Symp Soc. Exp Biol 45: 117-127; de Lange et al., (1995) Curr Top Microbiol Immunol 197: 57-75; Hamilton et al. (1995) Curr Top Microbiol Immunol 197: 77-89; Finnegan et al., (1996) Proc Nati Acad Sci USA 93: 8449-8454; Uhlmann and A. Pepan (1990), Chem. Rev. 90: 543; P. D. Cook (1991), Anti-Cancer Drug Design 6: 585; J. Goodchild, Bioconjugate Chem. 1 (1990) 165; and S. L. Beaucage and R. P.

Iyer (1993), Tetrahedron 49: 6123; and F. Eckstein, Ed. (1991), Oligonucleotides and Analogues- Practical Approach, IRL Press.

Catalytic RNA molecules or ribozymes can also be used to inhibit the expression of the ACC synthase genes. It is possible to design ribozymes that specifically pair with virtually any target RNA and segment the phosphodiester backbone in a specific location, in order to functionally inactivate the target RNA. When carrying out this segmentation, the ribosame is not itself altered, and thus it is able to recycle the segmentation of other molecules. The inclusion of ribosome sequences within antisense RNA confers activity of RNA segmentation in them, thus increasing the activity of the constructions. A number of classes of ribosomes have been identified. For example, a ribosome class is derived from a small number of circular RNAs that are capable of self-segmentation and replication in plants. The RNAs can replicate either alone (R? As viroide) or with a helper virus (R? As satellites). Examples of R? A include R? As of the avocado sunspot viroid and the satellite RNAs of the tobacco angular spot virus, lucerne transient fringe virus, tobacco mottled virus, nodular nodiflorum solano virus and spotted cloverleaf virus . The design and use of ribosimals specific for RNA has been described. See, for example, Haseloff et al. (1988) Nature, 334: 585-591. Another method to inactivate an ACC synthase gene by inhibiting expression is by sense suppression. The introduction of expression cassettes in which a nucleic acid is configured in sense orientation with respect to the promoter has been shown to be an effective means by which to block the transcription of a desired target gene. See, for example, Napoli et al. (1990), The Plant Cell 2: 279-289, U.S. Patent Nos. 5,034,323, 5,231,020, and 5,283, 184. Isolated or recombinant plants that include one or more inactivated ACC synthase genes as well. they can be produced by using RNA interference silencing (R? Ai), which can also be called posttranscriptional gene silencing (PTGS) or cosuppression. In the context of this invention, "RNA silencing" (also called RNAi or RNA-mediated interference) refers to any mechanism through which the presence of a single-stranded R? A or, typically, double-stranded R? a cell results in the inhibition of the expression of a target gene comprising a sequence identical or almost identical to that of R? A, including, but not limited to, interference from R? A, repression of the translation of a mR? A target transcribed from the target gene without alteration of mRNA stability and transcriptional silencing (eg, histone acetylation and etchocromatic formation leading to inhibition of target mRNA transcription). In "RNA interference" the presence of single-stranded or double-stranded RNA in the cell leads to endonucleolytic cleavage and then degradation of the target mRNA. In a modality, a transgene (for example, a sequence and / or subsequence of an ACC synthase gene or coding sequence) is introduced into a plant cell to inactivate one or more ACC synthase genes by silencing or interfering with R? A (R? Ai). For example, a sequence or subsequence includes a small subsequence, for example, of about 21-25 bases in length (with, for example, at least 80%, at least 90%, or approximately 100% identity to one or more than the subsequences of the ACC synthase gene), a larger subsequence, for example, about 25-100 or about 100-2000 (or about 200-1500, about 250-1000, etc.) bases in length (with at least a region of about 21-25 bases in length with at least 80%, at least 90%, or 100% identity to one or more subsequences of the ACC synthase gene), and / or the entire coding sequence or gene . In one embodiment, a transgene includes a region in the sequence or subsequences that is approximately 21-25 bases in length with at least 80%, at least 90%, or approximately 100% identity to the ACC synthase gene or coding sequence. The use of RNAi for the inhibition of gene expression in a number of cell types (including, for example, plant cells) and organisms, for example, by the expression of a hairpin RNA (stem-spiral) or the two strands of an interfering RNA, for example, as is well described in the literature, as are the methods for determining the appropriate R? A (s) for targeting a target gene, eg, an ACC synthase gene, and to generate such interfering RNAs. For example, RNA interference is described, for example, in the US patent publications and applications 20020173478, 20020162126, and 20020182223 in Cogoni and Macino (2000) "Post-transcriptionnal gene silencing across kingdoms" Genes Dev., 10: 638-643; Guru T. (2000), "A silence that speaks volumes"? Ature 404: 804-808; Hammond and collaborators, (2001), "Post-transcriptional Gene Silencing by Double-stranded RNA"? Ature Rev. Gen. 2: 110-119; ? apoli et al., (1990) "Introduction of a chalcone synthase gene into Petunia results in reversible co-suppression of homologous genes in trans." Plant Cell 2: 279-289; Jorgensen et al., (1996), "Chalcone synthase cosuppression phenotypes in petunia flowers: comparison of sense vs. antisense constructs and single-copy vs. complex T-DNA sequences "Plant Mol. Biol., 31: 957-973; Hannon G. J. (2002)" RNA interference "Nature., Jul 11; 418 (6894): 244-51; Ueda R. (2001) "RNAi: a new technology in the post-genomic sequencing era" J Neurogenet.; 15 (3-4): 193-204; Ullu and collaborators (2002) "RNA interference: advances and questions" Philos Trans R Soc Lond B Biol Sci. Jan 29; 357 (1417): 65-70; Waterhouse et al., (1998) Proc Nati Acad Sci USA 95: 133959-13964; Sch id et al. (2002) "Combinatorial RNAi: a method for evaluating the functions of gene families in Drosophila" Trends Neurosci. Feb; 25 (2): 71-4; Zeng et al. (2003) "MicroRNAs and small interfering RNAs can inhibit mRNA expression by similar mechanisms" Proc. Nati Acad. Sci. USA 100: 9779-9784; Doench et al. (2003) "siRNAs can function as miRNAs" Genes & Dev. 17: 438-442; Bartel and Bartel (2003) "MicroRNAs: At the root of plant development?" Plant Physiology 132: 709-717; Schwarz and Zamore (2002) "Why do miRNAs Uve in the miRNPl" Genes & Dev. 16: 1025-1031; Tang et al. (2003) "A biochemical framework for RNA silencing in plants" Genes & Dev. 17: 49-63; Meister et al. (2004) "Sequence-specific inhibition of microRNA-and siRNA-induced RNA silencing" R? A 10: 544-550; ? elson and collaborators (2003) "The microRNA world: Small is mighty" Trends Biochem. Sci. 28: 534-540; Dykxhoorn et al. (2003) "Killing the month senger: Short RNAs that silence gene expression "Nature Reviews Molec. and Cell Biol. 4: 457-467; McManus and Sharp (2002)" Gene silencing in mammals by small interfering RNAs "Nature Reviews Genetics 3: 737-747; Hutvagner and Za ore (2002) "RNAi: Nature abhors a double strand" Curr Opin Genet & Dev 200: 225-232; and Agami (2002) "RNAi and related mechanis7ns and their potential use for therapy" Curr Opin Chem Biol 6: 829-834. The sequence (s) or subsequence (s) of ACC synthase polynucleotide expressed to induce RNAi can be expressed, for example, under the control of a constitutive promoter, an inducible promoter, a tissue-specific promoter. The expression of a tissue-specific promoter may be advantageous in certain embodiments. For example, the expression of a leaf-specific promoter can inactivate one or more ACC synthase genes in the leaf, producing a green permanence phenotype, without activating the ACC synthase genes in the root (which can decrease the tolerance of the flood). Similarly, the expression of an anther-specific promoter can inactivate one or more ACC synthase genes in the anther, producing a male sterility phenotype, without inactivating the ACC synthase genes in the rest of the plant. Such methods are optionally combined, for example, to inactivate one or more ACC synthase genes in both leaves and anthers. Transposons The one or more ACC synthase genes can also be inactivated by, for example, inactivating the transposon-based gene. In one embodiment, the inactivation step comprises producing one or more mutations in an ACC synthase gene sequence, wherein the one or more mutations in the ACC synthase gene sequence comprises one or more transposon sensations, in order to inactivate the one or more ACC synthase gene compared to a corresponding control plant. For example, the one or more mutations comprises a homozygous interruption in the one or more ACC synthase gene or the one or more mutations comprise a heterozygous interruption in the one or more ACC synthase gene or a combination of both homozygous interruptions and heterozygous interruptions if interrupts more than one ACC synthase gene. The transposons were first identified in corn by Barbara McClintock in the late 1940s. The Mutator family of transposable elements, for example, transposable elements of Mutator de Robertson (Mu) are typically used in plants, for example, maize, mutagenesis of the gene , because they are present in high number of copies (10-100) and are inserted preferentially in and around the genes. The transposable elements can be classified by categories into two broad classes based on their mode of transposition. These are designated Class I and Class II; both have applications as mutagens and as delivery vectors. Class I transposable elements are transposed by an RNa intermediary and use reverse transcriptases, that is, they are retrograde. There are at least three types of Class I transposable elements, for example, retrotransposons, retroposons, elements similar to SINE. Retrotransposons typically contain LTRs, and genes that encode viral coat proteins (gag) and reverse transcriptase RnasaH, integrase and polymerase genes (pol) Numerous retrotransposons have been described in plant species. Such retrotransposons mobilize and translocalize via an RNA intermediate in a reaction catalyzed by reverse transcriptase and RNAse H encoded by the transposon. Examples are found in the Tyl-ccpiay Ty3-gypsy groups as well as in the SINE-like and LINE-like classifications. A more detailed discussion can be found in Kumar and Bennetzen (1999) Plant Retrotransposons in Annual Review of Genetics 33: 479.

In addition, the transposable elements of DNA such as Ac, Taml and En / Spm are also found in a variety of plant species and can be used in the invention. Transposons (and IS elements) are common tools for introducing mutations into plant cells.

These mobile genetic elements are delivered to the cells, for example, through a sexual cross, the transposition is selected and the resulting insertion mutants are classified, for example for a phenotype of interest. Plants comprising interrupted ACC synthase genes can be introduced into other plants by crossing isolated or recombinant plants with an uninterrupted plant, for example, by a sexual cross. Any of a number of standard breeding techniques can be used, depending on the species that is crossed. The location of a TN within a genome of a recombinant isolated plant can be determined by known methods, for example, the sequencing of flanking regions as described herein. For example, a PCR reaction of the plant can be used to amplify the sequence, and then it can be diagnostically sequenced to confirm its origin. Optionally, the insertion mutants are classified for a desired phenotype, such as the inhibition of the expression or activity of the ACC synthase, inhibition or reduced production of ethylene, potential for green permanence, etc. compared to a control plant. TILLING TILLING can also be used to inactivate one or more ACC synthase genes. TILLING is Targeting Induced Local Lesions IN Genomics. See, for example, McCallum et al., (2000), "Targeting Induced Local Lesions IN Geyaomics (TILLING) for Plant Functional Genomics" Plant Physiology 123: 439-442; McCallum et al., (2000) "Targeted screening for induced mutations" Nature Biotechnology 18: 455-457; and, Colbert et al, (2001) "High-Throughput Screening for Induced Point Mutations" Plant Physiology 126: 480-484. TILLING combines high-density point mutations with rapid sensitive detection of mutations. Typically, ethylmethanesulfonate (EMS) is used to mutagenize the plant seed. EMS rents guanine, which typically leads to parentage. For example, the seeds are soaked in a solution of approximately 10-20 mM EMS for about 10 to 20 hours; the seeds are washed and then sown. The plants of this generation are known as Ml. The Ml plants are then self-fertilized. Mutations that are present in the cells that make up the reproductive tissues are inherited by the next generation (M2). Typically, M2 plants are classified for mutation in the desired gene and / or specific phenotypes. For example, DNA from M2 plants is accumulated and mutations in an ACC synthase gene are detected by detection of heteroduplex formation. Typically, the DNA is prepared from each M2 plant and accumulated. The desired ACC synthase gene is amplified by PCR. The accumulated sample is then denatured and tempered to allow the formation of heteroduplexes. If a mutation is present in one of the plants; The PCR products will be of two types: wild type and mutant. Accumulations that include heteroduplexes are identified by separating the PCR reaction, for example, by Liquid Chromatography of High Denaturing Performance (DPHPLC). DPHPLC detects mismatches between the heteroduplexes created by melting and annealing the heteroallelic DNA. Chromatography is performed while the DNA is heated. The heteroduplexes have lower thermal stability and form melting bubbles resulting in faster movement in the chromatography column. When heteroduplexes are present in addition to the expected homoduplexes, a double peak is observed. As a result, the accumulations that carry the mutation in an ACC synthase gene are identified. The individual DNA of the plants that make up the accumulated population selected can then be identified and sequenced. Optionally, the plant that possesses a desired mutation in an ACC synthase can be crossed with other plants to remove antecedent mutations.

Other mutagenic methods can also be used to introduce mutations in an ACC synthase gene. The methods for introducing genetic mutations into plant genes and the selection of plants with desired attributes are well known. For example, the seeds or other plant material can be treated with a mutagenic chemical according to standard techniques. Such chemical substances include, but are not limited to, the following: diethyl sulfate, ethylene imine and N-nitroso-N-ethylurea. Alternatively, ionizing radiation from sources such as X-rays or gamma rays can be used. Other detection methods for detecting mutations in an ACC synthase gene may be employed, for example, capillary electrophoresis (e.g., constant denaturing capillary electrophoresis and single-strand conformational polymorphism). In another example, heteroduplexes can be detected using the mismatch repair enzymology (eg celery CEL I endonuclease). CEL I recognizes unequalization and segment exactly on the 3 'side of the unequalization. The precise base position of unequalization can be determined by cutting with the mismatch repair enzyme followed by, for example, denaturing gel electrophoresis. See, for example, Oleykowski et al., (1998) "Mutation detection using a novel plant endonuclease "Nucleic Acid Res. 26: 4597-4602; and Colbert et al., (2001)" High -Throughput Screening for Induced Point Mutations "Plant Physiology 126: 480-484 The plant containing the ACC synthase gene mutated can be crossed with other plants to introduce the mutation into another plant.This can be done using standard breeding techniques.Regulation Homologous Homologous recombination can also be used to inactivate one or more ACC synthase genes. Homologous recombination has been demonstrated in See, for example, Puchta et al. (1994), Experientia 50: 277-284, Swoboda et al. (1994), EMBO J. 13: 484-489, Offringa et al. (1993), Proc. Nati. Acad. Sci. USA 90: 7346-7350; Kempin et al. (1997) Nature 389: 802-803; Terada et al., (2002) "Efficient gene targeting by homologous recombination in rice" Nature Biotechnology, 20 (10): 1030-1034. Homologous recombination can be used to induce modifications of the targeted gene by specifically targeting an ACC synthase gene in vivo. Mutations in selected portions of an ACC synthase gene sequence (including upstream region 5 ', downstream 3' and intragenic regions) such as those provided in the present they are made in vitro and introduced into the desired plant using standard techniques. The mutated gene will interact with the Wild type ACC target synthase gene in a manner such that homologous recombination and targeted replacement of the wild-type gene will occur in transgenic plants, resulting in the suppression of ACC synthase activity. METHODS FOR MODULATING THE POTENTIAL OF GREEN PERMANENCE IN A PLANT The methods for modulating the potential for green permanence in plants are also characteristics of the invention. The ability to introduce different degrees of green permanence potential in plants offers a simple flexible procedure to introduce this attribute and a specific way for the purpose: for example, the introduction of a strong green permanence attribute for the improved grain filling or for silage in areas with longer or drier growing seasons against the introduction of a moderate green retention attribute for the corkage in areas with shorter growing seasons. In addition, the green permanency potential of a plant of the invention may include, for example, (a) a reduction in the production of at least one specific mRNA of ACC synthase; (b) a reduction in the production of an ACC synthase; (c) a reduction in ethylene production; (d) a delay in the senescence of the leaf; (e) an increase in drought resistance; (f) an increased time in maintaining the photosynthetic activity; (g) increased perspiration; (h) an increased stomatal conductance; (i) an assimilation of C02 increased; (j) an increased assimilation maintenance of C02; or (k) any combination of (a) - (j); compared to a corresponding control plant and the like. For example, a method of the invention may include: a) selecting at least one ACC synthase gene to mutate, to thereby provide at least one desired ACC synthase gene; b) introducing a mutant form of at least one desired ACC synthase gene in the plant and c) expressing the mutant form, in order to modulate the potential for green permanence in the plant. Plants produced by such methods are also a feature of the invention. The degree of green permanence potential introduced into a plant can be determined by a number of factors, for example, which ACC synthase gene is selected, whether the mutant gene member is present in a heterozygous or homozygous state, or by the number of members of this family who are inactivated, by a combination of two or more of such factors. In one embodiment, the selection of at least one ACC synthase gene comprises determining a degree (eg, weak (eg, ACS2), moderate or strong (eg, ACSd)) of desired green residence potential. For example, the ACS2 lines show a weak green permanence phenotype, such as a senescence delay of about one week. The ACC6 lines show a strong green permanence phenotype (for example, the leaf's oldness is delayed approximately 2-3 weeks or more). The ACS7 lines also show a strong green permanence phenotype (for example, the leaf's oldness is delayed approximately 2-3 weeks or more). For example, the ACC synthase gene is selected to encode a specific ACC synthase, such as, SEQ ID NOY (pACS2), SEQ ID N0: 8 (pACSd), SEQ ID NO: 9 (pAC7), or SEQ ED NO: 11 (pCCRAl78R). In one embodiment, two or more ACC synthase genes are interrupted, for example, ACS2 and ACSd), for example, to produce a strong green permanence phenotype. In other modalities, three or more ACC synthase genes are interrupted. Once the desired ACC synthase gene is selected, a mutant form of the ACC synthase gene is introduced into a plant. In certain modalities, the mutant form is introduced by Agrobacterium-mediated transfer, incorporation, bombardment of microprojectiles, homologous recombination, or a sexual cross. In certain modalities the mutant form includes, for example, a mutation heterozygous for at least one ACC synthase gene, a mutation homozygous in at least one ACC synthase gene, or a homozygous mutation or combination or heterozygous mutation if selected in an ACC synthase gene. In another embodiment, the mutant form includes a subsequence of at least one desired ACC synthase gene in an antisense, sense or silencing or RNA interference configuration. The expression of the mutant form of the ACC synthase gene or the result of the mutant form can be determined in a number of ways. For example, detection of expression products is performed either qualitatively (presence or absence of one or more products of interest) or quantitatively (by monitoring the level of expression of one or more products of interest). In one embodiment, the expression product is a product of RNA expression. The invention optionally includes monitoring a level of expression of a nucleic acid or polypeptide, as mentioned herein for the detection of ACC synthase in a plant or in a population of plants. Monitoring of ethylene ACC levels can also be used for the detection of inhibition of the expression or activity of a mutant form of the ACC synthase gene. In addition to increasing the tolerance to stress by drought in plants of the invention compared with a plant of control, another important aspect of the invention is that the highest planting density of plants of the invention may be possible, leading to increased yield per acre of corn. Most of the yield increase per acre of corn for the last century has come from increased tolerance to stacking, which is a stress in, for example, corn. Methods for modulating stress, for example, increasing tolerance for stacking in a plant are also a feature of the invention. For example, a method of the invention may include: a) selecting at least one ACC synthase gene to mutate, in order to thereby provide at least one desired ACC synthase gene; b) introducing a mutant form of at least one desired ACC synthase gene into the plant; and c) express the mutant form, in order to modulate the stress in the plant. Plants produced by such methods are also a feature of the invention. When ethylene production is reduced in a plant by a mutant form of a desired ACC synthase gene, the plant does not perceive piling. Thus, the plants of the invention can be planted in densities higher than what is currently practiced by farmers. In another aspect, the inactivation of one or more ACC synthase genes as discussed herein may influence the response to disease or pathogen attack.

METHODS FOR MODULATING STERILITY IN A PLANT Methods for modulating sterility, for example, male or female sterility, in plants are also characteristics of the invention. The ability to introduce male or female sterility into plants allows the rapid production of sterile female and male lines, for example, for use in commercial breeding programs, for example, for hybrid seed and production, where cross-pollination is desired. The inactive plants of ACC synthase, particularly inactive ACS6 and inactive double ACS2 / ACS6 have been observed to drop less dust than wild-type plants, suggesting the interruption of ethylene production as a novel means to modulate the sterility of the plant . For example, the method of the invention may include: a) selecting at least one ACC synthase gene to mutate, to thereby provide at least one desired ACC synthase gene; b) introducing a mutant form of at least one desired ACC synthase gene into the plant; and c) expressing the mutant form, in order to modulate the sterility in the plant. Plants produced by such methods are also a feature of the invention. Essentially all the characteristics mentioned in the above apply to this modality also, as relevant, for example, with respect to the number of ACC synthase and interrupted genes, techniques for introducing the mutant form of the ACC synthase gene into the plant, polynucleotide constructs and the like. In a modality class, at least one gene ACC synthase is interrupted by the insertion of a transposon, by a point mutation, or by the constitutive expression of a transgene comprising an ACC synthase polynucleotide in an antisense, sense or silencing or RNA interference configuration. Such lines can be propagated by exogenously providing ethylene, for example, by spraying the plants at an appropriate stage of development with 2-chloroethylphosphonic acid (CEPA), which decomposes in water to produce ethylene. In another class of embodiments, the at least one ACC synthase gene is interrupted by the expression of a transgene comprising an ACC synthase polynucleotide in an antisense, sense or silencing or interference A? an inducible promoter, such that sterility can be induced and / or repressed as desired. In yet another class of embodiments, at least one ACC synthase gene is interrupted by the expression of a transgene comprising an ACC synthase polynucleotide in a antisense, sense or silencing or RNA interference configuration under the control of a tissue-specific promoter, eg, a specific producer of anther to produce male sterile plants. Again, if necessary, such lines can be propagated by providing exogenous ethylene (for example, by spraying with CEPA). CLASSIFICATION / CHARACTERIZATION OF PLANTS OR CELLS OF PLANTS OF THE INVENTION The plants of this invention can be classified and / or characterized either genotypically; biochemically, phenotypically or a combination of two or more of these to determine the presence, absence and / or expression (eg, amount, modulation, such as a decrease or increase compared to a control cell, and the like) of a polynucleotide of the invention, the presence, absence, expression and / or enzymatic activity of a polypeptide of the invention, modulation of the green residence potential, modulation of the stacking and / or modulation of ethylene production. See, for example, Figure 19. Genotyping can be performed by any of a number of well-known techniques, including PCR amplification of genomic nucleic acid sequences and hybridization of acid sequences genomic nucleic or nucleic acid sequences expressed with specific labeled probes (e.g., Southern blot, Northern blot, dot blot or slit, etc.). For example, the Maize Attribute Utility System (TUSC), developed by Pioneer Irbid Int., Is a powerful PCR-based classification strategy to identify Mu transposon insertions in specific genes and the need for an observable phenotype. The system uses, for example, TIR-PCR in which one PCR primer is derived from the target gene and the other (Mu-TIR) from the terminal inverted repeat (TIR) region of Mu. Using these primers in accumulated DNA PCR reactions of a large population of Mu-containing plants, successful amplification is identified by Southern hybridization using the target gene as the probe. The classification of the individuals within a positive accumulation is then performed to identify the candidate line that contains the insertion of a Mu element in the target gene. In order to determine whether evidence of insertion is limited to somatic cells or is present in the germline (and therefore represents an inheritable change), the progeny of a candidate are optionally subjected to the same PCR analysis / Southern hybridization used in the original classification.

Biochemical analysis can also be performed to detect, for example, the presence, absence or modulation (eg, decrease or increase) of protein production (eg, by ELISAs, Westhern stains, etc.) the presence and / or the amount of ethylene produced and the like. For example, the expressed polypeptides can be recovered or purified from isolated or recombinant cell cultures by any of a number of methods well known in the art, including the precipitation of ammonium sulfate or ethanol, acid extraction, cationic ion exchange chromatography. , chromatography with phosphocellulose, hydrophobic interaction chromatography, affinity chromatography (for example, using any of the labeling systems mentioned herein) hydroxyapatite chromatography, lectin chromatography. The steps of protein refolding can be used, as desired, to complete the mature protein configuration. Finally, high performance liquid chromatography (HPLC) can be used in the final purification stages. In addition to the references mentioned in the above a variety of purification steps are well known in the art, including, for example, those set forth in Sandana (1997) Bioseparation of Proteins, Academic Press, Inc .; and Bollag et al. (1996) Protein Methods, 2nd Edition Wiley-Liss, NY; Walker (1996) The Protein Protocols Handbook Humana Press, NJ, Harris and Angal (1990) Protein Purification Applications: A Practical Approach IRL Press at Oxford, Oxford, England; Harris and Angal Protein Purification Methods: A Practical Approach IRL Press at Oxford, Oxford, England; Scopes (1993) Protein Purification: Principies and Practice 3rd Edition Springer Verlag, NY; Janson and Ryden (1998) Protein Purification: Principies, High Resolution Methods and Applications, Second Edition Wiley-VCH, NY; and Walker (1998) Protein Protocols on CD-ROM Humana Press, NJ. Chemicals, for example, ethylene, ACC, etc., can be recovered and analyzed from cell extracts. For example, the internal concentrations of ACC can be analyzed by gas chromatography-mass spectroscopy, in extracts of acidic plants such as ethylene after decomposition in alkaline hypochlorite solution, etc. The ethylene concentration can be determined by, for example, gas chromatography-mass spectroscopy, etc. See, for example, Nagahama, K., Ogawa, T., Fujii, T., Tazaki, M., Tañase, S., Morino, Y. and Fukuda, H. (1991) "Purification and properties of an ethylene forming enzyme from Pseudomonas syringae "J. Gen. Microbiol. 137: 2281-2286. For example, ethylene can be measured with a gas chromatograph equipped with, for example, a column based on alumina (such as a capillary column HP-PLOT A1203) and a flame ionization detector. Phenotypic analysis includes, for example, analyzing changes in chemical composition (for example, as described under biochemical analyzes), morphology or physiological properties of the plant. For example, morphological changes may include, but are not limited to, increased green dwell material, a delay in leaf senescence, an increase in drought resistance, an increase in stacking resistance, etc. Physiological properties may include, for example, increased sustained photosynthesis, increased transpiration, increased stomatal conductance. Assimilation of increased C02, longer maintenance of assimilation of C02, etc. A variety of tests can be used to monitor the potential for green permanence. For example, the assays include, but are not limited to, visual inspection, monitoring of photosynthesis measurements and measurement levels of chlorophyll, DNA, RNA and / or protein content of, for example, leaves. PLANTS OF THE INVENTION Plant cells of the invention include, but are not limited to, meristem cells, Type I callus, Type II and Type III, immature embryos and cells. gaméticas such as microspores, pollen, sperm, and ovule. In certain embodiments, the plant cells of the invention are dicotyledonous or monocotyledonous. A regenerated plant of the plant cell (s) of the invention is also a feature of the invention. In one embodiment, the plant cell is in a plant, for example a hybrid plant, comprising a phenotype of green residence potential. In another embodiment, the plant cell is in a plant comprising a sterility phenotype, eg male sterility phenotype. Through a series of reproduction manipulations, the interrupted ACC synthase gene can be moved from one plant line to another plant line. For example, the hybrid plant can be produced by sexual crossing of a plant comprising interruption of one or more ACC synthase genes and a control plant. The inactive plant cells are also a feature of the invention. In a first aspect, the invention provides an isolated or recombinant inactive plant cell comprising at least one interruption in the at least one endogenous ACC synthase gene (eg, a nucleic acid sequence, or complement thereof, which comprises, for example, at least about 70%, at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 99%, about 99.5% or more, a sequence identity of SEQ ID NO: 1 (gACS2 ), SEQ ID NO: 2 (gACSd) or SEQ ID NO: 3 (gACS7)). The disruption inhibits the expression or activity of at least one ACC synthase protein compared to a corresponding control plant cell that lacks the interruption. In one embodiment, in at least one endogenous ACC synthase gene comprises two or more endogenous ACC synthase genes. In another embodiment, in at least one endogenous ACC synthase gene comprises three or more endogenous ACC synthase genes. In certain embodiments, the at least one interruption results in the production of reduced ethylene by the inactive plant cell as compared to the control plant cell. In one aspect of the invention, the disruption of an ACC synthase gene in a plant cell comprises one or more transposons, where one or more transposons are in at least one endogenous ACC synthase gene. In another aspect, the interruption includes one or more point mutations in at least one endogenous ACC synthase gene. Optionally, the interruption is a homozygous interruption in at least one ACC synthase gene. Alternatively, the interruption is a heterozygous interruption in at least one ACC synthase gene.

In certain embodiments, more than one ACC synthase gene is involved and there is more than one interruption, which may include homozygous interruptions, heterozygous interruptions and a combination of homozygous interruptions and heterozygous interruptions. See also the sections entitled in this "Transposons and TILLING". In another embodiment, the interruption of an ACC synthase gene occurs by inhibiting the expression of the ACC synthase gene. For example, an inactive plant cell is produced by introducing at least one polynucleotide sequence comprising an ACC synthase nucleic acid sequence as subsequent thereto, into a plant cell, such that at least one sequence of The polynucleotide is linked to the promoter in a sense or antisense orientation. The polynucleotide sequence comprises, for example, at least about 70%, at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95% , at least about 99%, about 99.5% or more sequence identities to SEQ ID NO: l (gACS2), SEQ ID NO: 2 (gACSd), SEQ ID NOY (gACS7), SEQ ID NOY (cACS2), SEQ ID NOY (CACS6), SEQ ID NO: 6 (cACS7), or SEQ ID NO: 10 (CCRA178R), or a subsequence thereof or a complement thereto. For example, the inactive plant cell is may be produced by introducing at least one polynucleotide sequence comprising one or more subsequences and an ACC synthase nucleic acid sequence configured for interferent RNA silencing. The polynucleotide optionally comprises a vector, expression cassette or the like. In another aspect, the inactive plant cell is produced by homologous recombination. See also the sections entitled "Antisense, Sense, Squelch or RNA Interference Configurations" and "Homologous Recombination". Inactive plants comprising a phenotype of green residence potential is a feature of the invention. Typically, the green permanence potential phenotype in the active plant results from an interruption of at least one endogenous synthase ACC gene. In one embodiment, the interruption comprises one or more transposons and the disruption inhibits the expression or activity of at least one ACC synthase protein compared to a corresponding control plant. In another embodiment, the interruption comprises one or more point mutations and inhibits the expression or activity of at least one ACC synthase protein compared to a corresponding control. In certain embodiments, in at least one endogenous ACC synthase gene comprises a nucleic acid sequence, or complement thereof, comprising, for example,less about 70%, at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 99%, about 99.5% or more of sequence identity to SEQ ID NO: l (gACS2), SEQ ID NO: 2 (gACSd), or SEQ ID NOY gACS7), or complement thereof. In certain modalities the inactive plant is a hybrid plant. The invention also features inactive plants comprising a transgenic plant with a green permanence potential phenotype. For example, a transgenic plant of the invention includes a green permanence potential phenotype that results from at least one introduced transgene that inhibits ethylene synthesis, wherein in at least one introduced transgene it comprises a nucleic acid sequence encoding at least one ACC synthase or subsequence thereof, the nucleic acid sequence comprising, for example, at least about 70%, at least about 75%, at least about 80%, at least about 85% , at least about 90%, at least about 95%, at least about 99%, about 99.5% or more, of sequence identity of SEQ ID NO: 1 (gACS2), SEQ ID NO: 2 (gACSd), SEQ ID NOY (gACS7), SEQ ID NO: 4 (cACS2), SEQ ID NO: 5 (cACSd), SEQ ID NO: 6 (cACS7) or SEQ ID NO: 10 (CCRA178R), or a subsequence thereof, or a complement thereof, and modifies a level of expression or activity with at least one ACC synthase. Typically, the configuration is a configuration of sense, antisense or silencing or RNA interference. A transgenic plant of the invention may also include a green permanence potential phenotype that results from at least one introduced transgene that inhibits ethylene synthesis, wherein at least one introduced transgene comprises a nucleic acid sequence encoding sub-sequences of at least one ACC synthase, the at least one ACC synthase comprising, for example, at least about 70%, at least about 75%, at least about 80%, at least about 85%, so less about 90%, at least about 95%, at least about 99%, about 99.5% or more, of sequence identity to SEQ ID NOY (pACS2), SEQ ID NO: 8 (pACS6), SEQ ID NO: 9 (pACS7), or SEQ ID NO: l (pCCRAl78R), or a conservative variation thereof, and is in a configuration of RNA silencing or interference and modifies a level of expression or activity of the at least one ACC synthase. In one aspect, the transgene optionally comprises the tissue-specific promoter or an inducible promoter (eg, a leaf-specific promoter, a drought-inducible promoter, or the like).

The invention also characterizes inactive plants having a sterility phenotype, for example, a male or female sterility phenotype. Thus, one class of embodiments provides an inactive plant comprising a male sterility phenotype resulting from at least one interruption or at least one endogenous ACC synthase gene. The disruption inhibits the expression or activity of at least one ACC synthase protein compared to a corresponding control plant. For example, ACS2, ACSd and ACS7 can be interrupted, individually or any combination (for example, ACS6, or ACS2 and ACSd). Typically, the at least one interruption results in the reduced ethylene production of the inactive plant as compared to the control plant. In one embodiment, the at least one interruption comprises one or more transposons in at least one endogenous ACC synthase gene. In another embodiment, the at least one interruption comprises one or more point mutations in at least one endogenous ACC synthase gene. In another embodiment, the at least one interruption is introduced into the inactive plant by introducing at least one polynucleotide sequence comprising one or more subsequences of an ACC synthase nucleic acid sequence configured for RNA silencing and interference (or alternatively a sense or antisense configuration). How is it mentioned, the polynucleotide sequence is optionally under the control of a tissue-specific inducible promoter (eg, anther-specific). In one embodiment, the male sterility phenotype comprises the reduced fall of pollen by the inactive plant as compared to the control plant. For example, the inactive plant can drop at most 50%, 25%, 10%, 5%, or 1% of both pollen and the control plant, or can drop non-detectable pollen. The invention also features inactive plants comprising a transgenic plant as a male sterility phenotype. For example, a transgenic plant of the invention includes a male sterility phenotype that results from at least one introduced transgene that inhibits ethylene synthesis, wherein at least one introduced transgene comprises a nucleic acid sequence encoding at least one ACC synthase or subsequence thereof, the nucleic acid sequence comprising, for example, at least about 70%, at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 99%, about 99.5% sequence identity to SEQ ID NO: l (gACS2), SEQ ID NOY (gACSd), SEQ ID NOY (gACS7), SEQ ID NO: 4 (cACS2), SEQ ID NO: 5 (cACSd), SEQ ID NO: 6 (cACS7) or SEQ ID NO: 10 (CCRA178R), or a subsequence thereof, or a complement thereof, and modifies an expression level or activity of at least one ACC synthase. Typically, the configuration is a sense, antisense or silencing or RNA interference configuration. As mentioned, the transgene optionally comprises a tissue-specific promoter (e.g., an anther-specific promoter) or an inducible promoter. Essentially any plant can be used in the methods and compositions of the invention. Such species include but are not restricted to members of the families: Poaceae (formerly Graminae, including Zea mays (corn), rye, triticale, barley, millet, rice, wheat, oats, etc.); Leguminosae (including peas, beans, lentils, peanuts, sweet potato seed, cowpeas, hock, soybeans, clover, alfalfa, lupine, peas, lotus, clover, glycine, vetch, etc.); Compositae (the largest family of vascular plants, including at least 1,000 genera, including important cash crops such as sunflower) and Rosaciae (including raspberry, apricot, almond, peach, rose), as well as walnut plants (including, walnut, pecan, hazelnut, etc.), forest trees (including Pinus, Quercus, Pseutotsuga, Sequoia, Populus, etc.), and other common crop plants (for example, cotton, sorghum, meadow grass, tomato, potato, pepper, broccoli, cabbage, etc.) Additional plants, as well as those specified in the above, include plants of the genera: Acamptoclados, Achnatherum, Achnella, Acroceras, Aegilops, Aegopgon, Agroelymus, Agrohordeum, Agropogon, Agropyron, Agrosi tanion, Agrostis, Aira, Allolepis, Alloteropsis, Alopeeurus, Amblyopyrum, Ammophile, Ampelodesmos, Amphibromus, Amphicarpum, Amphilophis, Anastrophus, Anatherum, Andropogron, Anemathele, Aneurolepidium, Anisantha, Anthaenantia, Anthephora, Anthochloa, Anthoxanthum, Apera, Apluda, Archtagrostis, Arctophila, Argillochloa, Aristida, Arrhenatherum, Arthraxon, Arthrostylidium, Arundinaria, Arundinella, Arundo, Aspris, Atheropogon, Oats (for example, oats), Avenella, Avenochloa, Avenula, Axonopus, Bambusa, Beckmannia, Blepharidachne, Blepharoneuron, Bothriochloa, Bouteloua, Brachiaria, Brachyelytrum, Brachypodium, Briza, Brizopyrum, Bromelica, Bromopsis, Bromus, Buchloe, Bulbilis, Calamagrostis, Calamovilfa, Campulosus, Capriola, Catabrosa, Catapodium, Cathestecum, Cenchropsis, Cenchrus, Centotheca , Ceratochloa, Chaetochloa, Chasmanthi um, Chimo nobambusa, Chionochloa, Chloris, Chondrosum, Chrysopon, Chusquea, Cinna, Cladoraphis, Coelorachis, Coix, Coleanthus, Colpodium, Coridochloa, Cornucopiae, Cortaderia, Corynephorus, Cottea, Cri tesion, Crypsis, Ctenium, Cutandia, Cylindropyrum, Cymbopogon, Cynodon, Cynosurus, Cytrococcum, Dactylis, Dactyloctenium, Danthonia, Dasyochloa, Dasyprum, Davyella, Dendr ocal amu s, Deschampela, Desmazeria, Deyeuxia, Diarina, Diarrhena, Dichanthelium, Dichanthi um , Dichelachne, Diectomus, Digitaria, Dimeria, Dimorpostachys, Dinebra, Diplachne, Dissantheliu, Dissochondrus, Distichlis, Drepanostaclzymn, Dupoa, Dupontia, Echinochloa, Ectospina, Ehrharta, Eleusifze, Elyhordeum, Elyleymus, Elymordeum, Elymus, Elyonurus, Elysi tanion, Elytesion, Elytrigia, Enneapogon, Enteropogon, Epicampes, Eragrostis, Eremochloa, Eremopoa, Eremopyrum, Erianthus, Erikorrhea, Erichloa , Eriochrysis, Erioneuron, Euchlaena, Euclasta, Eulalia, Eulalíopsis, Eustachys, Fargesia, Festuca, Festulolium, Fingerhuthia, Fluminia, Garnotia, Gastridium, Gaudinia, Gigantechloa, Glyceria, Graphephorum, Gymnopogon, Gynerium, Hackelochloa, Hainardia, Hakonechloa, Haynaldia, Heleochloa , Helictotrichoyz, Hemarthria, Hesperochloa, Hesperostipa, Heteropogon, Hibanobambusa, Hierochloe, Hilaria, Holcus, Homalocenchrus, Hordeum (for example, barley), Hydrochloa, Hymenachne, Hyparrhenia, Hypogynium, Hystrix, Ichnanthus, Imperata, Indocalamus, Isachne, Ischaemurn, Ixophorus , Koeleria, Korycarpus, Lagurus, Lamarckia, Lasiacis, Leersia, Leptochloa, Leptochloopsis, Leptocoryphium, Leptoloma, Leptogon, Lepturus, Lerchenfeldia, Leucopoa, Leymostachys, Leymus, Limnodea, Li thaelafne, Lolium, Lophochlaena, Lophochloa, Lophopyrum, Ludolfia, Luziola, Lycurus, Lygeum, Maltea, Manisuris, Megastachya, Melica, Melinis, Mibora, Microchloa, Microlaena, Microstegium, Mílium, Miscanthus, Mnesi thea, Molinia , Monanthochloe, Monerma, Monroa, Muhlenbergia, Nardus, Nassella, Nazia, Neeragrostis, Neoschischkinia, Neostapfia, Neyraudia, Nothoholcus, Olyra, Opizia, Oplismenus, Orcuttia, Oryza (for example, rice), Oryzopsis, Otatea, Oxytenanthera, Paniculary, Panicum , Pappophorum, Parapholis, Pascopyrum, Paspalidium, Paspalum, Pennisetum (for example, millet), Phalaris, Phalaroides, Phanopyrum, Pharus, Phippsia, Phleum, Pholiurus, Phragmi tes, Phyllostachys, Piptatherum, Piptochaetium, Pleioblastus, Pleopogon, Pleuraphis, Pleuropogon, Poa, Podagrostis, Polypogon, Polytrias, Psathyrostachys, Pseudelymus, Pseudoroegneria, Pseudosasa, Ptilagrostis, Puccinellia, Pucciphippsia, Redfieldia, Reimaria, Reimarochloa, Rhaphis, Rhombolytrum, Rhynchelytrum, Roegneria, Rostraria, Rottboellia, Rytilix, Saccharum, Sacciolepis, Sasa, Sasaella, Sasamorpha, Savastana, Schedonnardus, Schisrnus, Schizachne, Schizachyrium, Schizostachyum, Sclerochloa, Scleropoa, Scleropogon, Scolochloa, Scribneria, Sécale (for example, rye), Semiarundinaria, Sesleria, Setaria, Shibataea, Sieglingia, Sinarundinaria, Sinobambusa, Sinocalamus, Sitanion, Sorghastrum, Sorghum, Spartina, Sphenopholis, Spodiopogon, Sporobolus, Stapfia, Steinchisma, Stenotaphrum, Stipa, Stipagrostis, Stiporyzopsis, Swallenia, Syntherisma, Taeniatherum, Terrellia, Terrelymus, Thamnocalamus, Themeda, Thinopyrum, Thuarea, Thysanolaena, Torresia, Torreyochloa, Trachynia, Trachypogon, Tragus, Trichachne, Trichloris, Tricholaena, Trichoneura, Tridens, Triodia, Triplasis, Tripogott, Tripsacutn, Trisetobromus, Trisetum, Triticosecale, Triticum (for example, wheat), Tuctoria, Unióla, Urachne, Uralepis, Urochloa, Vahlodea, Valota, Vaseyochloa, Ventenata, Vetiveria, Vilfa, Vulpia, Willkofnmia, Yushanía, Zea (for example, corn), Zizania, Zizaniopsis and Zoysia. TRANSFORMATION OF PLANTS The nucleic acid sequence constructs of the invention (for example isolated nucleic acids, recombinant expression cassettes, etc.) can be introduced into plant cells, either in culture or in the organs of plants, by a variety of conventional techniques. For example, the techniques include, but are not limited to, infection, transduction, transfection, transvection and transformation. Constructs of nucleic acid sequences can be introduced alone or with other polynucleotides. Other such polynucleotides can be introduced independently, co-introduced, or introduced in a manner linked to the polynucleotides of the invention. The techniques for transforming a wide variety of higher plant species are well known and described in the technical and scientific literature. See, for example, Payne et al., (1992) Plant Cell and Tissue Culture in Liquid Systems John Wiley & Sons, Inc. New York, NY (Payne); Gamborg and Phillips (eds) (1995) Plant Cell, Tissue and Organ Culture; Fundamental Methods Springer Lab Manual, Springer-Verlag (Berlin Heidelberg New York) (Gamborg); Croy, (ed.) (1993) Plant Molecular Biology Bios Scientific Publishers, Oxford, U. K; Jones (ed) (1995) Plant Gene Transfer and Expression Protocols-Methods in Molecular Biology, Volume 49 Humana Press Towata NJ and as well as others, etc., likewise, for example, Weising et al., (1988) Ann. Rev. Genet. 22: 421. See, also, WO 95/06128 entitled "Fertile, Transgenic Maize Plants and Methods for Their Production" published on March 2, 1995. Numerous prospects for the establishment of transformable protoplasts from a variety of plant types and subsequent transformation of Cultured protoplasts are available in the art and are incorporated herein by reference. For example, see, Hashimoto and collaborators, (1990) Plant Physiol. 93: 857; Fowke and Constabel (eds) (1994) Plant Protoplasts; Saunders et al., (1993) Applications of Plant In Vitro Technology Symposium, UPM 16- 18; and Lyznik et al., (1991) BioTechniques 10: 295, each of which is incorporated herein by reference. Numerous methods are available in the art for performing the transformation and expression of chloroplasts (eg, Daniell et al., (1998) Nature Biotechnology 16: 346; O'Neill et al., (1993) The Plant Journal 3: 729; Maliga ( 1993) TIBTECH 11: 1). For example, nucleic acid sequences can be introduced directly into the genomic DNA of a plant cell using techniques such as electroporation, PEG poration, particle bombardment, silicon fiber delivery or microinjection of plant cell protoplasts or embryogenic callus. or the nucleic acid sequence constructions can be introduced directly into the plant tissue using ballistic methods, such as bombardment of particles. Exemplary particles include, but are not limited to, tungsten, gold, platinum and the like. Alternatively, constructs of nucleic acid sequences can be introduced by infection of cells with viral vectors, or by combining nucleic acid constructs with suitable T-DNA framework regions and introduced into a conventional host vector of Agrobacterium tumefaciens. The virulence functions of the Agrobacterium host will direct the insertion of the construction and the adjacent marker in the DNA from the plant cell when the plant cell is infected by bacteria. See, U.S. Patent No. 5,591,616. Microinjection techniques are known in the art and well described in the scientific and patent literature (see, eg, Jones (ed) (1995) Plant Gene Transfer and Expression Protocols-Methods in Molecular Biology, Volume 49 Humana Press Towata NJ and as well as others). The insertion of constructions of nucleic acid sequence constructions using the polyethylene glycol precipitation is described in Paszkowski et al. (1984) EMBO J 3: 2717. Electroporation techniques are described in Fromm Y, (1985) Proc Nat'l Acad Sci USA 82: 5824.

Ballistic transformation techniques are described in Klein et al., (1987) Nature 327: 70; and Weeks et al., Plant Physiol 102: 1077 and by Tomes, D. et al., IN: Plant Cell, Tissue and Organ Culture: Fundamental Methods, Eds. O. L. Gamborg and G. C. Phillips, Chapter 8, pages 197-213 (1995). (See also Tomes et al, U.S. Patents Nos. 5,886,244, 6,258,999, 6,570,067, 5,879,918). I Viral vectors that are plant viruses can also be used to introduce polynucleotides of the invention into plants. Viruses are typically useful as vectors for expressing exogenous DNA sequences from a Transient way in plant hosts. In contrast to the agrobacterium-mediated transformation that results in the stable integration of the DNA sequence into the plant genome, the viral vectors are generally replicated and expressed without the need for chromosomal integration. Plant virus vectors offer a number of advantages, specifically: a) DNA copies of viral genomes can be easily manipulated E. coli and transcripts in vitro, where necessary, to produce copies of infectious RNA; b) Naked DNA, RNA or virus particles can be easily introduced into mechanically injured leaves of intact plants; c) high numbers of copies of viral genomes per cell resulting in high levels of expression of the introduced genes; d) common laboratory plant species as well as monocotyledonous and dicotyledonous species are easily infected by several strains of virus; e) infection of whole plants allows the sampling of repeated tissue from individual library clones; f) the recovery and purification of simple and rapid recombinant virus particles; and g) because replication occurs without chromosomal insertion, the expression is not subject to position effects. See, for example, Scholthof, Scholthof and Jackson, (1996) "Plant virus gene vectors for transient expression offoreign proteins in plants", Annu. Rev. of Phytopathol. 34: 299-323. Plant viruses cause a range of diseases, most commonly mottled damage to leaves, called mosaics. Other symptoms include necrosis, deformation, overgrowth and generalized yellowing or greening of the leaves. Plant viruses are known to infect each major food crop, as well as most species of horticultural interest. The host interval varies between viruses, with some viruses that infect a wide range of hosts (for example, the alfalfa mosaic virus infects more than 400 species in 50 plant families) while others have a reduced host range, sometimes limited to a few species, (for example barley yellow mosaic virus). Appropriate vectors may be selected based on the host used in the methods and compositions of the invention. In certain embodiments of the invention, a vector includes a plant virus, e.g., either RNA (single- or double-stranded) or DNA (single-stranded or double-stranded). Examples of such viruses include, but are not limited to, for example, an alfamovirus, a bromovirus, a capilovirus, a carlavirus, a carmovirus, a caulimovirus, a closterovirus, a comovirus, a cryptovirus, a cucumovirus, a diantovirus, a fabavirus , a fijivirus, a furovirus, a geminivirus, a hordeivirus, an ilarvirus, a luteovirus, a maclovirus, a chlorotic dwarf corn virus, a marafivirus, a necrovirus, a nepovirus, a yellow spotted parsnip virus, a pea-based mosaic virus, a potex virus, a potyvirus, a reovirus, a rhabdovirus, a sovere ovirus, a tenuivirus, a tobamovirus, a tobravirus, a tomato spotted wilt virus, a tombus virus, a thymus virus or the like. Typically, plant viruses encode multiple proteins required for initial infection, replication and systemic dispersion, for example coat proteins, helper factors, replicates and movement proteins. The nucleotide sequences that code for many of these proteins are knowledgeable subjects of public knowledge, and accessible through any of a number of databases, for example (Genbank: available on the world network site ncbi.nlm.nih .gov / genbank / or EMBL: available on the website of the global network ebi .ac.uk. embl /). Methods for the transformation of plants and plant cells using sequences derived from plant viruses include the direct transformation techniques described above related to the molecules of plants.

DNA, see for example, Jones, ed. (1995) Plant Gene Transfer and Expression Protocols, Humana Press, Totowa, NJ. Further, the viral sequences can be cloned adjacent to the T-DNA border sequences introduced via the Agrobacterium-mediated transformation or Agroinfection. Viral particles comprising plant virus vectors including polynucleotides of the invention can also be introduced by mechanical inoculation using techniques well known in the art; see, for example, Cunningham and Porter, eds. (1997) Methods in Biotechnology. Volume 3. Recombinant Proteins from Plants: Production and Isolation of Clinically Useful Compounds, for detailed protocols. Briefly, for experimental purposes, leaves of young plants are sprinkled with silicon carbide (carborundum), then inoculated with a solution of virally transcribed or encapsidated virus and gently rubbed. Large-scale adaptations to infect crop plants are also well known in the art, and typically involve mechanical maceration of leaves using a blinding or other mechanical implement, followed by localized spraying of viral suspensions, or spraying leaves with a regulated high-pressure virus / carborundum suspension. Any of these techniques mentioned in the above can be adapted to the vectors of the invention, and are useful for alternative applications depending on the choice of the plant virus and the host species, as well as the scale of the specific transformation application. In some embodiments, Agrobacterium-mediated transformation techniques are used to transfer the sequences or subsequences of ACC synthase of the invention to transgenic plants. The transformation mediated by Agrobacterium is widely used for the transformation of dicotyledons; However, certain monocotyledons can also be transformed by Agrobacterium For example, transformation by Agrobacterium of rice is described by Hiei et al. (1994) Plant J. 6: 271; U.S. Patent No. 5,187,073; U.S. Patent No. 5,591,616; Li and collaborators, (1991) Science in China 34:54; and Raineri and collaborators, (1990) Bio / Technology 8:33. Processed corn, barley, triticale and asparagus through the mediated transformation Agrobacterium has also been described (Xu et al. (1990) Chínese J Bot 2:81). The techniques of transformation mediated by Agrobacterium take advantage of the ability of the tumor-inducing plasmid (Ti) of A. tumefaciens to integrate into a genome of the plant cell to co-transfer a nucleic acid of interest in a plant cell. Typically, an expression vector is produced wherein the nucleic acid of interest, such as a nucleic acid of ACC synthase RNA configuration of the invention, is ligated into a plasmid autonomously replicating that also contains T-DNA sequences. The T-DNA sequences typically flank the nucleic acid of the expression cassette of interest and comprise the integration sequences of the plasmid. In addition to the expression cassette, T-DNA also typically includes a marker sequence, for example, antibiotic resistance genes. The plasmid with the T-DNA and the expression cassette are then transfected into Agrobacterium cells. Typically, for the effective transformation of plant cells, the A. tumefaciens bacterium also possesses the necessary vir regions on a plasmid or integrated into its chromosome. For a discussion of the Agrobacterium-mediated transformation, see, Firoozabady and Kuehnle, (1995) Plant Cell Tissue and Organ Culture Fundamental Methods, Gamborg and Phillips (eds.). The techniques of the transformation mediated by Agrobacterium tumefaciens are well described in the scientific literature. See, for example, Horsch et al., Science 233: 496-498 (1984) and Fraley et al., Proc. Nati Acad. Sci (USA) 80: 4803 (1983). Although the Agrobacterium is useful mainly in dicotyledons, certain monocots can be transformed by Agrobacterium um. For example, transformation by Agrobacterium of corn is described in U.S. Patent No. 5,550,318. Other methods of transfection or transformation include: (1) transformation by Agrobacterium rhizogenes (see, for example, Lichtenstein and Fuller In: Genetic Engineering, volume 6, PWJ Rigby, Ed., London, Academic Press, 1987, and Lichtenstein, CP, and Draper, J , In: DNA Cloning, Volume II, DM Glover, Ed., Oxford, IRI Press, 1985); Application PCT / US87 / 02512 (WO 88/02405 published April 7, 1988) describes the use of strain A4 of A. rhizogenes and its plasmid Ri together with the vectors of A. tumefaciens pARC8 or pARCld, (2) the lipid-mediated DNA uptake (see, for example, Freeman et al., Plant Cell Physiol., 25: 1353, 1984) and (3) the vortex formation method (see, for example, Kindle, Proc. Nat'l). Acad. Sci. (USA) 87: 1228, (1990) DNA can also be introduced into plants by transferring direct DNA into pollen as described by Zhou et al., Methods in Enzymology, 101: 433 (1983); D. Hess, Intern Rev. Cytol., 107: 367 (1987); Luo et al., Plant Mol. Biol. Repórter, 6: 165 (1988). The expression of polypeptide coding genes can be obtained by injecting the DNA into the reproductive organs of a plant as described by Pena et al., Nature 325: 274 (1987). The DNA can also be injected directly into the cells of immature embryos and the rehydration of dried embryos as described by Neuhaus et al., Theor. Appl. Genet., 75:30 (1987); and Benbrook et al., in Proceedings Bio Expo. 1986, Butterworth, Stoneham, Mass., Pages 27-54 (1986). A variety of plant viruses that can be employed as vectors are known in the art and include cauliflower mosaic virus (CaMV), geminivirus, bromine mosaic virus and tobacco mosaic virus. Other references describing suitable plant cell transformation methods include microinjection, Crossway et al., (1986) Biotechniques 4: 320-334; electroporation, Riggs et al., (1986) Proc. Nati Acad. Sci. USA 83: 5602-5606; transformation mediated by Agrobacterium um, see for example, Townsend et al., U.S. Patent No. 5,563,055; direct gene transfer, Paszkowski et al., (1984) EMBO J. 3: 2717-2722; and acceleration of ballistic particles, see for example, Sanford et al., U.S. Patent No. 4,945,050; Tomes et al., (1995) in Plant Cell, Tissue, and Organ Cul ture: Fundamental Methods, ed. Gamborg and Phillips (Springer-Verlag, Berlin); and McCabe et al., (1988) Biotechnology 6: 923-926. See also Weissinger et al., (1988) Annual Rev. Genet. 22: 421-477; Sanford et al., (1987) Particulate Science and Technology 5: 27-37 (onion); Christou et al., (1988) Plant Physiol. 87: 671-674 (soybean); McCabe et al. (1988) Bio / Technology 6: 923-926 (soybean); Datta et al., (1990) Biotechnology 8: 736-740 (rice); Klein et al. (1988) Proc. Nati Acad. Sci. USA 85: 4305-4309 (corn); Klein et al., (1988) Biotechnology 6: 559-563 (corn); Klein et al., (1988) Plant Physiol. 91: 440-444 (corn); Fromm et al., (1990) Biotechnology 8: 833-839; Hooydaas-Van Slogteren et al., (1984) Nature (London) 311: 763-764; Bytebier et al., (1987) Proc. Nati Acad. Sci. USA 84: 5345-5349 (Liliaceae); De Wet et al., (1985) in The Experimental Manipulation of Ovule Tissues, ed. G. P. Chapman et al., (Longman, New York), pages 197-209 (pollen); Kaeppler et al. (1990) Plant Cell Reports 9: 415-418; and Kaeppler et al., (1992) Theor. Appl. Genet 84: 560-566 (transformation mediated by whisker); D.

Halluin et al., (1992) Plant Cell 4: 1495-1505 (electroporation); Li et al. (1993) Plant Cell Reports 12: 250-255 and Christou et al. (1995) Annals of Botany 75: 407-413 (rice); Osjoda et al., (1996) Nature Biotechnology 14: 745-750 (maize via Agrobacterium tumefaciens); all of which are incorporated herein by reference. Regeneration of Isolated, Recombinant or Transgenic Plants Transformed plant cells that are derived by transformation techniques of plants and cells of isolated or recombinant plants, including those discussed above, can be cultured to regenerate an entire plant possessing the desired genotype (ie, inactive ACC synthase nucleic acid), and / or in this way the desired phenotype, for example, green permanence phenotype, sterility phenotype, stacking resistant phenotype, etc. The desired cells, which can be identified, for example, by selection or sorting, were cultured in the medium sustaining the regeneration. The cells are then allowed to mature in plants. For example, such regeneration techniques may depend on the manipulation of certain phytohormones in a tissue culture growth medium, which typically depends on a biocidal marker and / or herbicide that has been introduced together with the desired nucleotide sequences. Alternatively, classification can be performed to classify the inhibition of ACC synthase expression and / or activity, reduction in ethylene production conferred by the inactive ACC synthase nucleic acid sequence, etc. The regeneration of plants from cultured protoplast is described in Evans et al. (1983) Protoplasts Isolation and Culture, Handbook of Plant Cell Culture, pages 124-176, Macmillan Publishing Company, New York; Davey, (1983) Protoplasts, pages 12-29, Birkhauser, Basal 1983; Dale, Protoplasts (1983) pages 31-41, Birkhauser, Basel; and Binding (1985) Regeneration of Plants, Plant Protoplasts pages 21-73, CRC Press, Boca Raton. Regeneration can also be obtained from the plant callus, explant, organs or parts thereof. Such regeneration techniques are generally described in Klee et al., (1987) Ann Rev of Plant Phys 38: 467. See also, for example, Payne and Gamborg. For corn transformation and regeneration see, for example, U.S. Patent No. 5,736,369. The cells of plants transformed with a plant expression vector can be regenerated, for example, from individual cells, callus tissue or leaf discs according to the tissue culture techniques of standard plants. It is well known in the art that several cells, tissue organs of almost any plant can be successfully cultured to regenerate a whole plant. The regeneration of plants from cultured protoplasts is described in Evans et al., Protoplasts Isolation and Cul ture, Handbook of Plant Cell Culture, Macmillan Publishing Company, New, pages 124-176 (1983); and Binding, Regeneration of Plants, Plant Protoplasts, CRC Press, Boca Raton, pages 21-73 (1985). The regeneration of plants, which contain the foreign gene introduced by Agrobacterium from leaf explants it can be achieved as described by Horsch et al., Science, 227: 1229-1231 (1985). After transformation with Agrobacterium um, the explants are typically transferred to the selection medium. One aspect will understand that the means of selection depends on the selectable marker that is co-transfected in the explants. In this procedure, the transformants are grown in the presence of a selection agent and in a medium that induces the regeneration of shoots in plant species that is transformed as described by Fraley et al., Proc. Nat'l. Acad. Sci.

(E. U. A). , 80: 4803 (1983). This procedure typically produces suckers, for example, within two to four weeks, and these transformant shoots (which are typically, for example, approximately 1-2 cm in length) are then transferred to an appropriate root inducing medium containing the selective agent and an antibiotic. to prevent bacterial growth. Selective pressure is typically maintained in the middle of the root and shoots. Typically, the transformants will develop roots in approximately 1-2 weeks and will form seedlings. After the seedlings are about 3-5 cm in height, they are placed in sterile soil in fiber pots. Those skilled in the art will understand that different acclimation procedures are used to obtain transformed plants of different species. By example, after the development of the root and shoots, cuts, as well as somatic embryos of transformed plants, are transferred to the environment for the establishment of seedlings. For a description of the selection and regeneration of transformed plants, see, for example, Dodds and Roberts (1995) Experiments in Plant Tissue Culture, 3- Edition, Cambridge University Press. The transgenic plants of the present invention can be fertile or sterile. Regeneration can also be obtained from plant callus, explant, organs or parts thereof. Such regeneration techniques are generally described in Klee et al., Ann. Rev. of Plant Phys. 38: 467-486 (1987). The regeneration of plants from either individual plant protoplasts or several explants is well known in the art. See, for example, Methods for Plant Molecular Biology, A. Weissbach and H. Weissbach, eds., Academic Press, Inc., San Diego, California (1988). This process of regeneration and growth includes the stages of selection of transforming cells and shoots, rooting of the transformant shoots and growth of the seedlings in the soil. For the cultivation of corn cells and regeneration see generally, The Maize Handbook, Freeling and Walbot, Eds., Springer, New York (1994); Corn and Com Improvement, 3-edition, Sprague and Dudley Eds., American Society of Agronomy, Madison, Wisconsin (1988). One skilled artisan will recognize that after the recombinant expression cassette is stably incorporated into transgenic plants and confirmed that it is operable, it can be introduced into other plants by sexual cross.

Any of a number of standard breeding techniques can be used, depending on the species is crossed. In vegetatively propagated crops, mature transgenic plants can be propagated by cutting or by tissue culture techniques to produce multiple identical plants. The selection of desirable transgenics is made and new varieties are obtained and propagated vegetatively for commercial use. In propagated seed crops, mature transgenic plants can be auto-crossed to produce a homozygous inbred plant. The inbred plant produces seed containing the recently introduced heterologous nucleic acid. These seeds can be grown to produce plants that would produce the selected phenotype. Mature transgenic plants can also be crossed with other appropriate plants, generally other inbred or hybrid plants, including, for example, an isogenic non-transformed endomgamic plant. The parts obtained from the regenerated plant, such as flowers, seeds, leaves, branches, fruits and the like are included in the invention, as long as these parts comprise cells comprising the isolated nucleic acid of the present invention. Progeny and variants, and mutants of regenerated plants are also included within the scope of the invention, as long as these plants comprise the introduced nucleic acid sequences. Transgenic plants expressing the selectable marker can be classified for the transmission of the nucleic acid of the present invention by, for example, standard immunoblotting and DNA detection techniques. The transgenic lines are also typically evaluated on expression levels of the heterologous nucleic acid. The level expression of RNA can be determined initially to identify and quantify the expression of positive plants. Standard techniques for R? A analysis can be employed and include PCR amplification assays using oligonucleotide primers designed to amplify only heterologous RNA templates and solution hybridization assays using specific heterologous nucleic acid probes. The RNA positive plants can then be analyzed for protein expression by Western immunoblot analysis using the specifically reactive antibodies of the present invention. Further, In situ hybridization and immunocytochemistry according to standard protocols can be done using heterologous nucleic acid-specific polynucleotide probes and antibodies, respectively, to localize sites of expression within the transgenic tissue. Generally, a number of transgenic lines are usually classified for the incorporated nucleic acid to identify and select plants with the most appropriate expression profiles. Some embodiments comprise a transgenic plant that is homozygous for the added heterologous nucleic acid; that is, a transgenic plant that contains two added nucleic acid sequences, a gene in the same site on each chromosome of a pair of chromosomes. A homozygous transgenic plant can be obtained to sexually couple (self-reproduce) a heterozygous (aka hemizygous) transgenic plant containing a heterologous individual added nucleic acid, by germinating some of the seed produced and by analyzing the resulting plants produced for the altered expression of a polynucleotide of the present invention in relation to a control (i.e., native, non-transgenic) plant. The subsequent crossing to a plant of origin and the exogamic crossing with a non-transgenic plant, or with a transgenic plant for the same or another attribute or attributes are also contemplated. It is also expected that the transformed plants will be used in traditional breeding programs, including TOPCROSS pollination systems as disclosed in US 5,706,603 and US 5,704,160, the description of each of which is incorporated herein by reference. In addition to Berger, Ausubel and Sambrook, general references useful for cloning plant cells, culture and regeneration include Jones (ed) (1995) Plant Gene Transfer and Expression Protocols-Methods in Molecular Biology. Volume 49 Humana Press Towata NJ; Payne et al., (1992) Plant Cell and Tissue Culture in Liquid Systems John Wiley & Sons, Inc. New York, NY (Payne); and Gamborg and Phillips (eds) (1995) Plant Cell, Tissue and Organ Culture: Fundamental Methods Springer Lab Manual, Springer-Verlag (Berlin Heidelberg New York) (Gamborg). A variety of cell culture media are described in Atlas and Parks (eds) The Handbook of Microbiological Media (1993) CRC Press, Boca Raton, FL (Atlas). Additional information for the cultivation of plant cells is found in commercial literature available such as Life Science Research Cell Culture Catalog (1998) from Sigma-Aldrich, Inc. (St Louis, MO) (Sigma-LSRCCC) and, for example, the Plant Culture Catalog and supplement (1997) also from Sigma-Aldrich, Inc (St Louis, MO) (Sigma-PCCS). Additional details that consider The cultivation of plant cells is found in Croy, (ed.) (1993) Plant Molecular Biology Bios Scientific Publishers, Oxford, UK "STACKING" OF BUILDINGS AND ATTRIBUTES In certain embodiments, the nucleic acid sequences of the present invention can be used in combination ("stacked") with other polynucleotide sequences of interest in order to create plants with a desired phenotype. . The polynucleotides of the present invention can be stacked with any gene or combination of genes, and the combinations generated can include multiple copies of any one or more of the polynucleotides of interest. The desired combination can affect one or more attributes; that is, certain combinations can be created for the modulation of gene expression that affects the activity of ACC synthase and / or the production of ethylene. Other combinations may be designed to produce plants with a variety of desired attributes, including but not limited to desirable attributes for feeding animals such as high oil content genes (eg, US Patent No. 6,232,529); balanced amino acids (eg hordothionines (U.S. Patent Nos. 5,990,389; 5,885,801; 5,885,802; and 5,703,409); high-content barley of lysine ((Williamson et al., (1987) Eur. J. Biochem. 165: 99-106; and WO 98/20122); and high-methionine proteins (Pedersen et al., (1986) J. Biol. Chem 261: 6279; Kirihara et al. (1988) Gene 71: 359; and Musumura et al. (1989) Plant Mol. Biol. 12: 123)); increased digestibility (e.g., modified storage proteins (North American application serial number 10 / 053,410 filed November 7, 2001); and thioredoxin (North American application serial number 10 / 005,429 filed December 3, 2001) the descriptions of which are incorporated herein by reference.The polynucleotides of the present invention can also be stacked with desirable attributes for insect, disease or herbicide resistance (eg, Bacillus thuringiensis toxic proteins (U.S. Patent Nos. 5,366,892; 5,747,450; 5,737,514; 5723,756; 5,593,881; Geiser et al., (1986) Gene 48: 109), lectins (Van Damme et al., (1994) Plant Mol. Biol. 24: 825), fumonisin detoxification genes (North American patent No 5, 792, 931), genes for resistance to avirulence and disease (Jones et al., (1994) Science 266: 789; Martin et al., (1993) Scien 262: 1432; Mindrinos et al., (1994) Cell 78: 1089); acetolactate synthase (ALS) mutants that drive resistance to such herbicide such as mutations S4 and / or Hra; glutamine synthase inhibitors such as phosphinothricin or coarse (for example, the bar gene); and resistance to glyphosate (EPSPS gene)); and desirable attributes for processing or process products such as high oil content (eg, US Patent No. 6,232,529); modified oils (e.g., fatty acid desaturase genes (U.S. Patent No. 5,952,544; WO 94/11516)); modified starches (eg, 7? DPG pyrophosphorylases (AGPase), starch tape (SS), starch branching enzymes (SBE) and starch debranching enzymes (SDBE)); and polymers or bioplastics (for example, North American patent No. 5,602,321; beta-ketothiolase, polyhydroxybutyrate synthase and acetoacetyl-CoA reductase (Schubert et al., (1988) J. Bacteriol 170: 5837-5847) facilitate the expression of polyhydroxyalkanoates (PHAs)), the descriptions of which are incorporated herein by reference. Polynucleotides of the present invention could also be combined with polynucleotides that affect agronomic attributes such as male sterility (e.g., see U.S. Patent No. 5,583,210), stem strength, flowering time or attributes of transformation technology such such as regulation of the cell cycle or the direction of the gene (for example WO 99/61619, WO 00/17364, WO 99/25821), the descriptions of which are incorporated herein by reference. These stacked combinations can be created by any method, including but not limited to cross-breeding of plants by any conventional methodology or TopCross, or genetic transformation. If the attributes are stacked by genetically transforming the plants, the polynucleotide sequences of interest can be combined at any time and in any order. For example, a transgenic plant comprising one or more desired attributes can be used as the objective to introduce additional attributes by subsequent transformation. The attributes can be introduced simultaneously into a co-transformation protocol with the polynucleotides of interest provided by any combination of transformation cassettes. For example, if two sequences will be introduced, the two sequences can be contained in separate transformation cassettes (trans) or contained in the same transformation cassette (cis). The expression of the sequences of interest can be induced by the same promoter or by different promoters. In certain cases, it may be desirable to introduce a transformation cassette that will suppress the expression of a polynucleotide of interest. This can be done by any combination of other suppression cassettes or over-expression cassettes to generate the desired combination of attributes in the plant. USE IN REPRODUCTION METHODS The transformed plants of the invention can be used in a plant breeding program. The objective of plant reproduction is to combine, in a single variety or hybrid, several desirable attributes. For field crops, these attributes may include, for example, resistance to diseases and insects, tolerance to heat and drought, reduced time to maturity of the crop, higher yield and better agronomic quality. With the mechanical harvest of many crops, the uniformity of plant characteristics such as germination and establishment of the support, proportion of growth, maturity and plant and height of the ear is desirable. Traditional plant breeding is an important tool in developing new and improved cash crops. This invention comprises methods for producing a maize plant by crossing a first parental maize plant with a second parental maize plant wherein one or both of the parental maize plants is a transformed plant that exhibits a green permanence phenotype, a phenotype of sterility, a stacking resistance phenotype or the like, as described is described herein. The techniques of reproduction of known plants in the art and used in a breeding program of corn plants include, but are not limited to, recurrent selection, volume selection, mass selection, subsequent crossing, pedigree reproduction, open pollination reproduction, increased selection of the polymorphism of restriction fragment length, increased selection of the genetic marker, double aploids and transformation. Frequently combinations of these techniques are used. The development of corn hybrids in a breeding program for maize plants requires, in general, the development of homozygous inbred lines, the crossing of these lines and the evaluation of the crosses. There are many analytical methods available to evaluate the outcome of a cross. The oldest and most traditional method of analysis is the observation of phenotypic attributes. Alternatively, the genotype of a plant can be examined. A genetic attribute that has been designed on a particular maize plant using transformation techniques can be moved on another line using traditional breeding techniques that are well known in plant breeding techniques. For example, a subsequent cross-over procedure is commonly used to move a transgene from a transformed corn plant to a line Inbred elite and the resulting progeny would then comprise the transgene (s). Also, if an inbred line was used for the transformation, then the transgenic plants could be crossed to a different inbred line in order to produce a hybrid transgenic maize plant. As used herein, "crossing" can refer to a simple X-by-Y cross, or the subsequent cross-over process, depending on the context. The development of a corn hybrid in a maize plant breeding program involves three stages: (1) the selection of plants from several accumulations of plasma germ for initial breeding crosses; (2) the self-reproduction of the plants selected from breeding crosses for several generations to produce a series of inbred lines, which, while different from each other, actually reproduce and are highly uniform; and (3) the crossing of the selected inbred lines with different inbred lines to produce the hybrids. During the process of reproduction of inbreeding in corn, the vigor of the lines decreases. The vigor is restored when two different inbred lines are crossed to produce the hybrid. An important consequence of homozygosity and homogeneity of inbred lines is that the hybrid created by crossing a defined pair of inbred lines will always be the same. Once that the inbred lines give a superior hybrid that is identified, the hybrid seed can be reproduced indefinitely while the homogeneity of the inbred origins is maintained. The transgenic plants of the present invention can be used to produce, for example, a single-cross hybrid, a three-way hybrid or a double-cross hybrid. A single-cross hybrid occurs when two inbred lines are crossed to produce the progeny Fl. A double cross hybrid is produced from four inbred lines crossed in pairs (A x B and C x D) and then the two hybrids Fl are crossed again (A x B) x (C x D). A three-way cross hybrid is produced from three inbred lines where two of the inbred lines are crossed (A x B) and then the resulting hybrid Fl is crossed with the third inbred line (A x B) x C. Much of the hybrid vigor and the uniformity exhibited by the hybrids Fl is lost in the next generation (F2). Consequently, the seed produced by the hybrids is consumed before planted. ANTIBODIES The polypeptides of the invention can be used to produce antibodies specific for the polypeptides, SEQ ID NOY SEQ ID NO: 9 and SEQ ID NO: 11, and conservative variants thereof. The specific antibodies for, for example, SEQ ID NO: 7-9 and 11, and polypeptides of related variants are useful, for example, for classification and identification purposes, for example, related to the activity, distribution and expression of ACC synthase. Antibodies specific for the polypeptides of the invention can be generated by methods well known in the art. Such antibodies may include, but are not limited to, polyclonal, monoclonal, chimeric, humanized, single chain, Fab fragments and fragments produced by an Fab expression library. The polypeptides do not require biological activity for the production of antibody. The full length polypeptide, subsequences, fragments or oligopeptides can be antigenic. Peptides used to induce specific antibodies typically have an amino acid sequence of at least about 10 amino acids, and often at least 15 or 20 amino acids. Short stretches of a polypeptide, for example, selected from SEQ ID NO: 7 SEQ ID NO: 9 and SEQ ID NO: 11, can be fused with another protein, such as the limpet hemocentesin and the antibody produced against the chimeric molecule. Numerous methods for producing polyclonal and monoclonal antibodies are known to those skilled in the art and can be adapted to produce antibodies specific for the polypeptides of the invention, for example, corresponding to SEQ ID NO: 7 SEQ ID NO: 9 and SEQ ID NO: 11. See, for example, Coligan (1991) Current Protocols in Immunology Wiley / Greene, NY; and Harlow and Lane (1989) Antibodies: A Laboratory Manual Cold Spring Harbor Press, NY; Stites et al. (Eds.) Basic and Clinical Immunology (4th edition) Lange Medical Publications, Los Altos, CA and references cited therein; Goding (1986) Monoclonal Antibodies: Principies and Practice (2- edition) Academic Press, New York, NY; Fundamental Immunology, for example, 4- Edition (or last), W. E. Paul (ed.), Raven Press, N. Y. (1998); and Kohler and Milstein (1975) Nature 256: 495-497. Other techniques suitable for the preparation of antibody include the selection of libraries of recombinant antibodies in phage or similar vectors. See, collaborators of Huse Y, (1989) Science 246: 1275-1281; and Ward, et al., (1989) Nature 341: 544-546. Specific monoclonal and polyclonal antibodies and antisera will usually bind to a KD of at least about 0.1 μM, preferably at least about 0.01 μM or better, and more typically preferably, 0.001 μM or better. EQUIPMENT TO MODULATE THE POTENTIAL OF GREEN PERMANENCE 0 ESTERILIDAD Certain embodiments of the invention may optionally be provided to a user as a team. For example, a kit of the invention may contain one or more nucleic acid, polypeptide, antibody, diagnostic nucleic acid or polypeptide, eg, antibody, set of probes, for example, as a cDNA microarray, one or more vectors and / or cell line described herein. More often, the equipment is packaged in a suitable container, the equipment typically also comprises one or more additional reagents, for example, substrates, labels, primers or the like to mark the expression products, tubes and / or other accessories, reagents to collect Samples, regulatory solutions, hybridization chambers, deck plates, etc. The kit optionally further comprises a set of instruction or user's manual detailing the preferred methods for using the components of the equipment for the discovery or application of the gene sets. When used according to the instructions, the equipment can be used, for example, to evaluate the expression or polymorphisms in a plant sample, for example, to evaluate ACC synthase, ethylene production, green permanence potential, resistance potential to stacking, sterility, etc. Alternatively, the equipment may be used in accordance with the instructions to use at least one sequence of ACC synthase polynucleotide to control the green permanency potential in a plant. As another example, a device for modulating sterility, eg, male sterility, in a plant includes a container that contains at least one polynucleotide sequence comprising a nucleic acid sequence, wherein the nucleic acid sequence is, for example, example, at least about 70%, at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 99% , approximately 99.5% or more, identical to SEQ ID NO: 1 (gACS2), SEQ ID NO: 2 (gACS6), SEQ ID NO: 3 (gACS7), SEQ ID NOY (CACS2), SEQ ID NO: 5 (cACSd), SEQ ID NO: 6 (cAC7) or SEQ ID NO: 10 (CCRA178R), or a consequence thereof, or a complement to the same The equipment optionally also includes instructional materials for the use of at least one polynucleotide sequence to control sterility, eg, male sterility, in a plant. OTHER NUCLEIC ACID AND PROTEIN TESTS In the context of the invention, nucleic acids and / or proteins are manipulated according to methods of "well-known molecular biology." Detailed protocols for numerous such procedures are described, example, in Ausubel et al., Current Protocols in Molecular Biology (supplemented until 2004) John Wiley & Sons, New York ("Ausubel"); Sambrook et al., Molecular Cloning-A Laboratory Manual (2- Edition), Volume 1-3, Cold Spring Harbor Laboratory, Cold Spring Harbor, New York, 1989 ("Sambrook") and Berger and Kimmel Guide to Molecular Cloning Techniques, Methods in Enzymology volume 152 Academic Press, Inc., San Diego, CA ("Berger"). In addition to the above references, protocols for in vitro amplification techniques, such as polymerase chain reaction (PCR), ligase chain reaction (LCR), Qβ-replicase amplification and other techniques mediated by RNA polymerase (eg, NASBA), useful for example, to amplify polynucleotides of the invention, is found in Mullis et al., (1987) U.S. Patent No. 4,683,202; PCR Protocols A Guide to Methods and Applications (Innis et al., Eds) Academic Press Inc. San Diego, CA (1990) ("Innis"); Arnheim and Levinson (1990) C & EN 36; The Journal Qf NIH Research (1991) 3:81; Kwoh et al., (1989) Proc Nati Acad Sci USA 86, 1173; Guatelli et al., (1990) Proc Nati Acad Sci USA 87: 1874; Lomell et al. (1989) J Clin Chem 35: 1826; Landegren et al. (1988) Science 241: 1077; Van Brunt (1990) Biotechnology 8: 291; Wu and Wallace (1989) Gene 4: 560; Barringer et al. (1990) Gene 89: 117 and Sooknanan and Malek (1995) Biotechnology 13: 563. Additional methods, useful for cloning nucleic acids in the context of the invention, include Wallace et al., In U.S. Patent No. 5,426,039. Improved methods for amplifying large nucleic acids by PCR are summarized in Cheng et al. Y, (1994) Nature 369: 684 and references therein. Certain polynucleotides of the invention can be synthesized using various solid phase strategies involving phosphoramidite coupling chemistry based on mononucleotide and / or trinucleotide. For example, nucleic acid sequences can be synthesized by the sequential addition of activated monomers and / or trimers to a lengthening polynucleotide chain. See, for example, Caruthers, M. H. et al., (1992) Meth Enzymol 211: 3. Instead of synthesizing the desired sequences, essentially any nucleic acid can be ordered from any variety of commercial sources, such as The Midland Certified Reagent Company ([email protected]) (Midland, Texas), The Great American Gene Company ( available on the World Wide Web Site at genco.com) (Ramona, CA), ExpressGen, Inc. (available from the World Network Site expressgen.com) (Chicago IL), Operon Technologies, Inc. (available from the Site of the World Network to operon.com) (Alameda CA) and many others.

EXAMPLES The following examples are offered to illustrate, but not to limit the claimed invention. EXAMPLE 1: INSULATION OF INACTIVATIONS OF CORN SYNTHESIS Because ethylene has been associated with the promotion of leaf senescence in some species, to introduce the potential for green permanence in, for example, maize, the inventors undertook to reduce the ethylene biosynthesis in corn leaves through the inactivation of ACC synthase genes. The family of the corn synthase ACC gene is composed of three members: ACS2, ACSd and ACS7. In order to isolate ethylene mutants, the inventors classified interruptions of each member of the ACC synthase gene family using the Maize Attribute Utility System (TUSC). To date, the inventors have determined the Mu insertion site for 8 mutant lines (three ACSd and five ACS2) by sequencing through the binding of Mu / ACC synthase. Five insertions were stably inherited; their positions are indicated in Figure 2, which also schematically represents ACS7. A pronounced green permanence phenotype was observed on the leaves of those plants in which a member of a single gene of the C.A family. was present in a heterozygous mutant state (see Figure 3, Panels A, B, C and D).

When present in a homozygous mutant state, an even more pronounced green permanence phenotype was observed (see Figure 4). In Figure 4, a leaf of wild type (left), inactivation of heterozygous ACC synthase (middle part) and inactivation of homozygous ACC synthase (right) was covered for 7 (days) in the dark. The leaves of inactive homozygous ACC synthase plants exhibited a larger green permanence attribute than leaves of heterozygous ACC synthase inactivation and exhibited a green permanence attribute substantially larger than the leaves of wild type plants. The degree of green permanence potential introduced was specific to members of the gene. Consequently, a strong green permanence attribute was introduced with the mutation of one member (eg, ACSd), whereas a less pronounced green permanence phenotype was introduced with the mutation of another member (eg, ACS2). Therefore, the degree of green permanence potential introduced into a line can be controlled, by which the mutant gene member is introduced, if the mutant gene member is present in a heterozygous or homozygous state, and by the number of members of this family that are inactivated (for example, ACS2 / ACS6 double mutants have a green permanence phenotype) strong) . The attributes associated with improved hybrid posture include resistance to stem rot and leaf pests, genetic stem resistance, short plant height and ear placement, and high potential for green permanence. Typically, the leaves follow a typical progression of initiation through expansion that eventually ends in old age. Carbon fixation capacity also increases during expansion and finally declines at low levels through old age. See, for example, Gay AP and Thomas H (1995) Leaf development in Loli um temulentum: photosynthesis in relation to growth and senescence. New Phytologist 130: 159-168. This is of particular relevance to cereal species where the potential yield is more dependent on the ability of the plant to fix carbon and store this carbon in the seed, mainly in the form of starch. Both the synchronization in which old age is initiated and the proportion in which it progresses can have a significant impact on the total carbon of a particular leaf that can ultimately contribute to a plant. See, for example, Thomas H and Howarth CJ (2000) Five ways to stay green. Journal of Experimental Botany 51: 329-337. This is particularly relevant for those crops where the potential yield is reduced by the conditions Adverse environmental conditions that induce premature senescence of the leaf. Green permanence is a general term used to describe a genotype whereby the senescence of the leaf (more mainly distinguished by the yellowing of the leaf associated with chlorophyll degradation) is retarded compared to a standard reference. See, for example, Thomas and Howarth, supra. In sorghum, several genotypes of green permanence have been identified that exhibit a delay in leaf senescence during grain filling and maturation. See, for example, Duncan RR, et al., (1981) Descriptive comparison of senescent and nofa-senesceyzt sorghum genotypes. Agronomy Journal 73: 849-853. In addition, under conditions of limited water availability, which normally hastens the leaf's senescence (for example, Rosenow DT and Clark LE (1981) Drought tolerance in sorghum, In: Loden HD, Wilkinson D, eds Proceedings of the 36th annual corn and sorghum industry research conference, 18-31), these genotypes retain more green area the leaf continues to fill the grain normally (for example, McBee GG et al., (1983) Effect of senescence and non-senescence on carbohydrates in sorghum during late kemel maturi and states. 23: 372-377; Rosenow DT et al., (1983) Drought-tolerant sorghum and cotton gennplasiiz, Agricultural Water Management 7: 207-222, and Borrell AK, Douglas ACL (1996) Maintaining green leaf area in grain sorghum increases yield to water-limited environment. In: Foale MA, Henzell RG, Kneipp JF, eds. Proceedings of the third Australian sorghum conference. Melbourne: Australian Institute of Agricultural Science, Occasional Publication No. 93). The green permanence phenotype has also been used as a selection criterion for the development of improved maize varieties, particularly with respect to the development of drought tolerance. See, for example, Russell WA (1991) Genetic improvement of maize yields. Advances in Agronomy 46: 245-298; and Bruce et al., (2002), Molecular and physiological approaches to maize improvenzert for drought tolerance, Journal of Experimental Botany, 53 (366): 13-25. Five fundamentally different types of green permanence have been described. See, for example, Thomas H and Smart CM (1993) Crops that stay green. Annals of Applied Biology 123: 193-219; and Thomas and Howarth, supra. In the green type A stay the initiation of the old age program is delayed, but then proceeds at a normal rate. In the green type B stay, while the initiation of the senescence program is unchanged, the progression is comparatively slower. In green Type C permanence, chlorophyll is retained although senescence (as determined by measurements of physiological function such as photosynthetic capacity) proceeds to a normal proportion. The green permanence of type D is more artificial in that the extermination of the leaf (that is, by freezing, boiling or drying) prevents the initiation of the senescence program thus halting the degradation of chlorophyll. In the green permanence of Type E, initial levels of chlorophyll are higher while the initiation and progression of leaf senescence are unchanged, thus giving the illusion of a relatively slower rate of progression. Type A and B are functional green permanences since the photosynthetic capacity is maintained together with the chlorophyll content and are the types associated with increased yield and tolerance to drought in sorghum. Despite the potential importance of this attribute, particularly the benefits associated with increased yield and tolerance to drought, very little progress has been made in understanding the biochemical, physiological or molecular basis for genetically determined green permanence. See, for example, Thomas and Howarth, supra. A number of environmental and physiological conditions have been shown to significantly alter the synchronization and progression of leaf senescence and may provide some insight into the basis for this attribute. Among environmental factors, light is probably the most significant and has been established by long time that leaf senescence can be induced in many plant species by placing leaves disjointed in the dark. See, for example, Weaver LM, Amasino RM (2001) Senescence is induced in individually darkened Arabidopsis leaves, but inhibited in whole darkened plants. Plant Physiology 127: 876-886. Limited availability of nutrients and water has also been shown to induce leaf senescence prematurely (eg, Rosenow DT, Quisenberry JE, Wendt CW, Clark LE (1983) Drougth-tolerant sorghum and cotton germplasm.) Agricultural Water Management 7: 207-222). Among the physiological determinants, growth regulators play a key role in directing the leaf's senescence program. Of particular relevance is the observation that the modification of cytokinin levels can significantly retard the senescence of the leaf. For example, plants transformed with isopentenyl transferase (ipt), an Agrobacterium gene that encodes a limiting stage of proportion in biosynthesis of cytokinin, when placed under the control of an inducible senescence promoter, resulted in a self-regulated cytokinin progression and a strong green permanence phenotype. See, for example, Gan S, Amasino RM (1995) Inhibition of leaf senescence by autoregulated production of cytokinin. Science 270: 1986-1988. Ethylene has also been implicated in the control of leaf senescence (for example, Davis KM and Grierson D (1989) Identification of cDNA clones for tomato (Lycopersicon esculentum Mili). mRNAs that accumulate during fruit ripening and leaf senescence in response to ethylene. Plant 179: 73-80) and plants impaired in the production or perception of ethylene also show a delay in leaf senescence (eg, Picton S et al., (1993) Alteredfruit ripening and leaf senescence in tomatoes expressing an antisense ethylene-forming enzyme transgene, The Plant Journal 3: 469-481, Grbic V and Bleeker AB (1995), Ethylene regulates the timing of leaf senescence in Arabidopsis, The Plant Journal 8: 95-102, and, John I et al., ( 1995) Delayed leaf senescence in ethylene-deficient ACC-oxidase antisense tomato plants: molecular and physiological analysis, The Plant Journal 7: 483-490), which can be phenotyped by the exogenous application of biosynthesis inhibitors and ethylene action ( example, Abeles FB et al., (1992) Ethylene in Plant Biology Academic Press, San Diego, CA). The identification and analysis of mutants in Arabidopsis and tomato that are deficient in the biosynthesis and perception of ethylene are valuable in establishing the important role that ethylene plays in the growth and development of the plant. The analysis of the mutant has also been in identifying and characterizing the transduction path of the ethylene signal. While many mutants of Ethylene have been identified in dicotyledonous plants (e.g., Arabidopsis and tomato), none of such mutants has been identified in monocots (e.g., rice, wheat and corn). Here the identification of maize mutants deficient in ACC synthase, the first enzyme in the ethylene biosynthetic pathway, is described. These mutants are critical in elucidating the regulatory functions that ethylene plays through cereal development as well as its function and regulatory responses to environmental stress. The knowledge gained from such mutant analyzes will increase understanding of the role of ethylene in the development of corn and will be relevant for other cereal crop species. The mutants were deficient in the production of ethylene and exhibited a green permanence phenotype. Green permanence was observed under normal growth conditions and after prolonged drought conditions that induced the premature onset of leaf senescence in wild type plants. In addition to maintaining chlorophyll during water stress, the leaves deficient in ACC synthase maintained the photosynthetic function and continued assimilating C02. Surprisingly, reducing the production of ethylene improved leaf function in all leaves under normal growth conditions and maintained a high level of function in plants stressed by drought even for those leaves in which senescence has not been induced in leaves of similar age of wild type plants. These findings indicate that ethylene can serve to regulate leaf function under normal growth conditions as well as in response to drought conditions. MATERIALS AND METHODS Cloning of Zea mays ACC synthase genes To facilitate the cloning of maize ACC gene synthase, primers were designed for highly conserved regions between multiple monocot species and dicotyledonous species using the sequence information currently available in GenBank. Initial PCR reactions were carried out on maize genomic DNA using the ACCF1 primers (ccagatgggcctcgccgagaac; SEQ ID NO: 12) and ACCl (gttggcgtagcagacgcggaacca; SEQ ID NO: 13) and revealed the three fragments of different sizes. All three fragments were sequenced and confirmed to be highly similar in sequence to other known ACC synthase genes. To obtain the complete genomic sequences for each of these genes, all three fragments were radiolabeled with dCTP using the Primea-Gene labeling system (Promega) and used to classify a genomic library (Stratagene) of corn EMBL3 (B73) from according to methods described in Sambrook, supra. Hybridization took Overnight at 30 ° C in buffer containing 5X SSPE, 5X Denhardt's, 50% formamide and 1% SDS. The spots were washed sequentially at 45 ° C in IX SSPE and 0.1X SSPE containing 1% SDS was exposed to film at -80 ° C with an intensifying screen.A total of 36 confluent plates (150 mm diameter) were The putative positive plates were subsequently directly classified by PCR using the above primers to identify which clones contained fragments corresponding to the three fragments initially identified.The PCR classification was carried out using HotStarTaq (Qiagen) .The reactions contained regulatory solution IX, 200 μM of each dNTP, 3 μM MgCl2, front and rear primer 0.25 μM, 1.25 U HotStarTaq 25U 1 μL of primary phage dilution (1/600 total in SM buffer) or a template in a total reaction volume of 25 μL The reaction conditions were as follows: 95 ° C / 15 min. (1 cycle); 95 ° C / 1 min, 62 ° C / 1 min, 72 ° C / 2 min (35 cycles); 72 ° C / 5 min (1 cycle). The samples were separated on a 1% agarose gel and the products were visualized after staining with ethidium bromide. All amplified fragments were also subjected to restriction analysis to identify other potential sequence-specific differences dependent on the size of the subsequence.

To facilitate sequencing of the remaining portions of these genes, the ACCF1 and ACCl primers were used in conjunction with specific primers for either the left arm (gacaaactgcgcaactcgtgaaaggt; SEQ ID NO: 14) or right arm (ctcgtccgagaataacgagtggatct; SEQ ID NO: 15) ) of the EMBL3 vector to amplify each half of the gene. Takara LA Taq (Panvera) was used to amplify the fragments due to the large size. The reactions contained 1 μl of phage dilution (1/600 total in SM buffer) and 2 μM of each primer (final concentration), regulatory solution IX (final concentration), 400 μM dNTP mixture (final concentration) and 1.25 U LA Taq in 25 μl of total volume. The reactions were carried out under the following conditions: 98 ° C / 1 min. (1 cycle); 98 ° C / 30 sec, 69 ° C / 15 min. (35 cycles); The amplified products were purified using the StrataPrep PCR purification kit (Stratagene) and sent to the sequencing facility at the University of Florida, Gainesville for direct sequencing. Identification of inactive ACC synthase mutants Maize has been proven to be a rich source of mutants, in part due to the presence of active or previously active transposable element systems within its genome. Depending precisely on the location of the insertion site in a gene, a transposon can partially or completely inactivate the expression of a gene. The inactivation of the gene may or may not have an observable phenotype depending on the amount of redundancy (ie the presence of multiple family members and the tissue specificity of the family members). The Maize Attribute Utility System (TUSC), developed by Pioneer Hybrid Int., Is a powerful PCR-based classification strategy for identifying Mu transposon insertions in specific genes without the need for an observable phenotype. This classification procedure is best suited for target genes that have previously been isolated from maize. The system uses TIR PCR in which one PCR primer is derived from the target gene and another (Mu-TIR) from the terminal inverted repeat region (TIR) Mu. Using these primers in accumulated DNA PCR reactions from a large population of plants containing Mu, successful amplification is identified by Southern hybridization using the target gene as the probe.

The classification of the individuals within a positive accumulation is then performed to identify the candidate line that contains insertion of a Mu element in the target gene.

In order to determine whether evidence of insertion is limited to somatic cells or is present in the germ line (thus represents an inheritable change), the progeny of a candidate undergo the same PCR / Southern hybridization analysis used in the classification original. A research effort was established to identify the inactive mutants in ethylene biosynthesis. To accomplish this, four primers (ACCFl, ccagatgggcctcgccgagaac, SEQ ID NO: 12, ACC-1, gttggcgtagcagacgcggaacca, SEQ ID NO: 13, ACC-C, cagttatgtgagggcacaccctacacacca, SEQ ID NO: 16, ACC-D, catcgaatgccacagctcgaacaacttc, SEQ ID NO : 17) specific to the corn ACC synthase genes discussed in the above were used to classify the Mu insertions in combination with the Mu-TIR primer (aagccaacgcca (a / T) cgcctc (c / T) atttcgt; SEQ ID N0: 18). The initial classification resulted in the putative identification of 19 separate lines carrying Mu insertions in the ACC corn synthase multigen family. The seed of each of these lines was planted and the DNA was extracted from the leaf of each individual. For the isolation of DNA, lcm "of seedling leaf was isolated from each plant and placed in a 1.5 ml centrifuge tube containing some sand.The samples were quickly frozen in liquid nitrogen and ground to a fine powder using a disposable disposer (Fisher Scientific) 600 μl of extraction buffer (100 mM Tris (pH 8.0), 50 mM EDTA, 200 mM NaCl, 1% SDS, 101 μl / ml β-mercaptoethanol) was added immediately and was mixed thoroughly, 700 μl of Phenol / Chloroform (1: 1) was added and the samples were centrifuged 10 min at 12,000 rpm. 500 μl of supernatant were removed to a new tube and the nucleic acid precipitated at -20 ° C after the addition of 1/10 vol of 3 M sodium acetate and 1 vol of isopropanol. The total nucleic acid was pelletized by centrifugation at 12,000 rpm, washed 3X with 75% ethanol and resuspended in 600 μl H20. The PCR classification was performed using HotStarTaq (Qiagen). The reactions contained buffer solution IX, 200 μM of each dNTP, MgmY 3mM, 0.25 μM of specific ACC synthase primer (ACCFl, ACC-1, ACC-C or ACC-D), 0.25 μM of Mu-specific primer (MuTIR), 0.25 μl of HotStarTaq and 1.5 μl of total nucleic acid as a template in a total reaction volume of 25 μl. The reaction conditions were as follows: 95 ° C / 15 min. (1 cycle); 95 ° C / 1 min, 62 ° C / 1 min, 72 ° C / 2 min (35 cycles); 72 ° C / 5 min (1 cycle). The PCR products were separated on a 1% agarose gel, visualized after staining with ethidium bromide and transferred to nylon membranes according to the methods described Sambrook Y et al., (1989). The Suthern stain analysis was performed as described above for the library classification except that the hybridization temperature was increased to 45 ° C. Seed BC1 (posterior cross 1) was planted from each of the 13 putative mutant lines and sorted by PCR / Southern analysis (as described precisely). From these lines, only 5 were found to be stably inherited. These five lines were crossed again 4 additional times to minimize the effects of the unrelated Mu inserts. The BC5 seed self-pollinated to generate homozygous null individuals. Individual homozygous null lines were identified by PCR using Takara LA Taq and the ACCF1 and ACC-1 primers. The reactions contained 1 μl of leaf DN, 2 μl of each primer (final concentration), regulatory solution IX (final concentration), 400 μM of dNTP mixture (final concentration) and 1.25 U LA Taq in 25 μl of total volume. The reactions were carried out under the following conditions: 98 ° C / 1 min. (1 cycle); 98 ° C / 30 sec, 69 ° C / 15 min. (35 cycles); 72 ° C / 10 min (1 cycle). PCR of wild-type B73 DNA using these primers and the conditions results in the amplification of three fragments of different size corresponding to the three identified genes. Individuals that are either wild type or heterozygous for one of the null insertion alleles exhibit this characteristic pattern while those that are homozygous for one of the null insertion alleles are losing subsequence corresponding to the gene in which the null insertion is located. insertion. To determine the exact location of Mu insertion site, the PCR products of each of these lines were amplified using either the ACCFl or ACC-1 primer in combination with the MuTIR primer. These fragments were then sequenced through the Mu / target binding using the Mu-TIR primer. The location of these Mu elements within each of the ACC synthase genes is shown in Figure 2. Protein Extraction For isolation of total protein, B73 leaves or mutant plants were harvested at the indicated times, frozen rapidly in nitrogen and ground into a fine powder. One ml of extraction buffer (20 mM HPES (pH 7.6), 100 mM KCl, 10% Glycerol) was added approximately 0.1 g of frozen powder and mixed thoroughly. The samples were centrifuged 10 min at 10,000 rpm, the supernatant was removed in a new one and the concentration was determined spectrophotometrically according to the methods of Bradford (1976). See, Bradford MM (1976) A rapid and sensitiv e rnethodfor the quantification of microgram quantities of protein utilizing the principie of protein-dye binding. Anal. Biochem. 72: 248-254. (See Figure 18). Extraction of chlorophyll The leaves were frozen in liquid nitrogen and ground to a fine powder. Samples of approximately 0.1 g were removed to a 1.5 ml tube and weighed. The Chlorophyll was extracted 5X with 1 ml (or 0.8 ml) of 80% acetone. The individual extractions were combined and the final volume was adjusted to 10 ml (or 15 ml) with additional 80% acetone. The chlorophyll content (a + b) was determined spectrophotometrically according to the methods of Ellburn (1994). See, Wellburn, A. R. (1994) The spectral determination of chlorophylls a and b, as well as total caretenoids using various solvents witla spectrophotometers of different resolution. J. Plant Physiol. 144: 307-313. (See Figure 17). Measurement of Photosynthesis Plants were grown in the field under normal conditions and drought stress. The normal plants were irrigated for eight hours a week. For drought-stressed plants, water was limited to approximately four hours per week during a period starting approximately one week before pollination and continuing through weeks after pollination. During the limited water availability period, drought-stressed plants showed visible signs of wilting and leaf curl. Transpiration, stomatal conductance and C02 ailation were determined with a portable TPS-1 photosynthesis system (PP Systems). Each leaf in a plant was measured in forty days after pollination. The values They represent an average of six determinations. See Figures 5 and 6. Purification of DNA and RNA For the isolation of total nucleic acid, B73 leaves were collected at desired times, quickly frozen in liquid nitrogen and ground to a fine powder. Ten ml of extraction buffer (100 mM Tris (pH 8.0), 50 mM EDTA, 200 mM NaCl, 1% SDS, 10 μl / L of β-mercaptoethanol) are added and mixed thoroughly until thawed. Ten ml of Phenol / Chloroform (1: 1, vol: vol) is added and mixed thoroughly. The samples are centrifuged 10 min at 8,000 rpm, the supernatant is removed in a new tube and the nucleic acid is precipitated at -20 ° C after the addition of 1/10 vol of 3 M sodium acetate and 1 vol isopropanol. The total nucleic acid is pelletized by centrifugation at 8,000 rpm and resuspended in 1 ml of TE. Half of the preparation is used for DNA purification and the remaining half is used for RNA purification. (Alternatively, DNA or total nucleic acids can be extracted 1 cm "from the seedling leaf, frozen rapidly in liquid nitrogen, and ground to a fine powder. 600 μl extraction buffer [100 mM Tris (pH 8.0 ), 50 mM EDTA, 200 mM NaCl, 1% SDS, 10 μl / ml of β-mercaptoethanol] are added and the sample is mixed. 700 μl of phenol / chloroform (1: 1) and centrifuged for 10 min at 12,000 rpm. The DNA is precipitated and resuspended in 600 μl of H20).

For DNA purification, 500 μg of DNase-free RNase is added to the tube and incubated at 37 ° C for 1 hour. After digestion of RNase, an equal volume of Phenol / Chloroform (1: 1, vol: vol) is added and mixed thoroughly. The samples are centrifuged 10 min at 10,000 rpm, the supernatant is removed to a new tube and the D? A precipitated -20 ° C after the addition 1/10 vol of 3 M sodium acetate and 1 vol of isopropanol. The D? A is resuspended in sterile water and the concentration is determined spectrophotometrically. To determine the integrity of D? A, 20 mg of D? A are separated on a 1.8% agarose gel and visualized after staining with ethidium bromide. The R? A is purified during 2 rounds of precipitation with LiCl2 according to the methods described by Sambrook et al., Supra. Real-time RT-PCR analysis Fifty μg of total R? A is treated with RQ1 of D? Asa (Promega) to assure that contaminating D? A is not present. Two μg of total R? A is directly used for the synthesis of cD? A using the OmniscriptRT equipment (Qiagen) with oligo-dT (20) as the primer. The analysis of transcript abundance is performed using the QuantiTect SYBR eGreen PCR equipment (Qiagen). The reactions containing 1 X buffer, 0.5 μl of the reverse transcription reaction (equivalent to 50 ng of total RNA) and 0.25 μM (final concentration) of front and back primers (see table 2 below) in a reaction volume total of 25 μl. TABLE 2 The reactions were carried out using an ABI PRISM 7700 sequence detection system under the following conditions: 95 ° C / 15 min. (1 cycle); 95 ° C / 30 sec, 62 ° C / 30 sec, 72 ° C / 2 min (50 cycles); 72 ° C / 5 min (1 cycle). Each gene is analyzed at a minimum of four times. All primer combinations are initially run and visualized on an agarose gel to confirm the presence of the individual product of the correct size. All amplification products are subcloned into the pGEM-T Easy system vector (Promega) for use in the generation of standard curves to facilitate the conversion of the expression data to a base of one copy / μg of RNA. Determination of ethylene Ethylene was measured from the second fully-extended leaf of the seedling leaves in the 4-leaf stage or from the 15 cm end of the leaves of plants 20, 30 or 40 days after pollination (DAP). The leaves were collected at the indicated times and allowed to recover for 2 hours before the ethylene collection, between wet paper towels. The leaves were placed in glass vials and covered with a rubber stopper. After an incubation of 3-4 hours, 0.9 mL of the upper space was sampled from each vial and the ethylene content was measured using a 6850 series gas chromatography system (Hewlett-Packard, Palo Alto, CA) equipped with a column capillary-based alumina from HP Plot (Agilent Technologies, Palo Alto, CA). The weight of fresh tissue was measured from each sample. Three replicates were measured and the average and standard deviation were reported. Western blot analysis B73 leaves were collected at the indicated times and ground in liquid nitrogen to a fine powder. One ml of extraction buffer [HEPES 20 mM (pH 7.6), 100 mM KCl KCl, 10% glycerol, 1 mM PMSF] was added approximately 0.1 g of frozen powder and mixed thoroughly. The cell debris was formed into pellets by centrifugation at 10,000 rpm for 10 min and the protein concentration was determined as described (Bradford, 1976). The antiserum highlighted against the large rice Rubisco subunit was obtained from Dr. Tadahiko Mae (Tohoku University, Sendai, Japan). The protein extracts were resolved using standard SDS-PAGE and the protein was transferred to a 0.22 μm nitrocellulose membrane by electro-stamping. After transfer, the membranes were blocked in 5% milk, 0.01% trimeral in TPBS (0.1% Tween 20, 7 mM NaCl, 0.27 mM KCl, 1 mM NaCHP04, 0.14 mM KYP04) followed by incubation with antibodies Primary diluted typically 1: 1000 to 1: 2000 in TPBS with 1% milk for 1.5 hours. The spots were then washed twice with TPBS and incubated with antibodies conjugated with goat anti-rabbit horseradish peroxidase (Southern Biotechnology Associates Inc.) diluted 1: 5000 to 1: 10,000 for 1 hr. The spots were washed twice with TPBS and the signal was typically detected between 1 to 15 min using chemiluminescence (Amersham Corp). RESULTS Identification of inactive ACC synthase mutants Three genes encoding ACC synthase were isolated from the inbred B73 and sequenced (see, for example, SEQ ID NO: 1-11). Two family members (ie, ACS2 and ACS7) are closely related (97% identity of amino acids) while the third gene (ie, ACS6) is considerably more divergent (54% and 53% amino acid identity with ACS2 and ACS7, respectively). An inverse genetic procedure was used to classify the transposon insertions in the family members of the ACC synthase gene (Bensen et al., (1995) Cloning and characterization of the maize Anl gene. Plant Cell 7: 75-84). 19 candidate lines were identified, 13 of which were confirmed by terminal inverted repeat (TIR) ~ PCR to harbor a Mu insertion in one of the three ACC synthase genes. Of these, 5 lines stably inherited the transposon in the first cross after B73 which was subsequently crossed 4 additional times to reduce the unwanted Mu attachments. The plants were then self-pollinated to generate homozygous null individuals that were identified by PCR using the ACCF1 and ACC-1 primers (see Methods). PCR amplification of the wild-type or heterozygous null mutant lines with these primers resulted in three fragments of different size corresponding to the three ACC synthase genes while the PCR amplification products to the homozygous null mutants lack the corresponding fragment to the mutant gene. The Mu insertion site for each mutant line was determined by sequencing through the Mu /? CC synthase binding using the .TIR primer (Figure 2).

Four of the five insertion lines contained a Mu in ACS2: one mutant contained one insertion in the third exon while the other three contained insertions in the fourth exon in unique positions (Figure 2). The fifth insertion line contained a Mu in ACSd in the second intron near the 3 'splice site. Quantitative real-time RT-PCR revealed that all three genes are expressed during the development of the corn leaf and confirmed that Mu insertions resulted in the loss of or a decrease in ACS expression. The insertions in ACS7 were identified in the first generation but were not inherited, suggesting that they were somatic mutants or that the expression of ACS7 is required for the development of the germ line. For a description of ACC synthase expression patterns during endosperm and embryo development, see Gallie and Young (2004). The ethylene biosynthetic and perception machinery is differen- tially expressed during elidospenn and embryo development Mol Gen Genomics 271: 267-281. Disruption of the ACS6 or ACS2 Gene Reduces Ethylene Synthesis The level of ethylene emission in maize leaves was increased as a function of leaf age (Figure 19 Panel C). In 20 DAP, the highest level of ethylene was observed in leaf 1 (the oldest surviving leaf) that by 30 DAP has progressed to leaf 3 and by 40 DAP (ie, core maturity) to leaves 4-5. To determine whether the Mu interruptions of ACSd or ACS2 described above reduced the ethylene emission, ethylene was measured from leaf 4 of the wild-type and mutant plants. The ethylene emission of the acs2 plants was about 55% of the wild-type plants, a level that was similar for all the alleles. mutants acs2 (Figure 19 Panels A-B). The ethylene emission of the acs6 plants was only 10% of that of the wild type plants (Figure 19 Panel B). The ethylene emission of the acs2 / acs 6 double mutant plants was similar to that of the acsß plants. These data suggest that the loss of ACSd expression results in a greater reduction in the ability of corn leaves to produce ethylene than the loss of ACS2 expression. Disruption of ACSd Confers a Green Permanence Phenotype A substantial increase in ethylene emission correlated with the appearance of visible signs of senescence in wild-type leaves, suggesting that ethylene can promote the entry of leaves into the program. senescence. If so, a delay in the senescence of the acs ß sheets, which produce significantly less ethylene, It would be expected. To test this possibility, homozygous (that is, acs6 / acs6), heterozygous (ie ACS6 / acs6) and wild type (ie ACS6 / ACS6) plats were grown in fields up to 50 days after pollination. In this stage, the older wild type leaves have aged, while the corresponding ACS6 / acs6 leaves only started to age and the acsß / acsß leaves remained green complemented. These observations suggest that the level of ethylene emission can determine the synchronization of the leaf's age. Senescence can also be induced after prolonged exposure to darkness. To determine if a reduction in ethylene emission can retard dark-induced senescence, leaves of adult plants were covered with covers to exclude light for two weeks. Leaves of younger plants (ie, 20 DAP) were used to ensure that age-related senescence will not occur during the course of the experiment and they remained in the plant. The corn cultivation in the greenhouse was also used to avoid any heating that could occur in the field as a consequence of the covering. After treatment of two weeks of darkness, the old age was observed for virtually the entire region of the wild-type leaves that were covered (the region covered by the cover is indicated by the transition different from yellow to green, Figure 4 on the left). The tip of the leaves ACSß / acsβ has been subjected to darkness-induced senescence but the rest of the covered region showed significantly less senescence (Figure 4 at the center). In contrast, the acsß / acsß sheets remained completely green (Figure 4 on the right). The degree of senescence correlated with the amount of ethylene produced by each, in which the ACSß / acsß sheets produced only 70% of the wild-type ethylene and the acsß / 'acsß sheets produced only 14.6% of the wild-type ethylene. These results suggest that ethylene mediates the onset of darkness-induced senescence such as natural senescence. They also indicate that the heterozygous ACSß / acsβ mutant with a loss of one copy of ACSß produces less ethylene and exhibit a weak green permanence phenotype similar to that observed for the acs2 mutant that also exhibited a moderate reduction (ie, 40%) in the emission of ethylene. To examine whether exogenous ACC could complement the acsß mutant and reverse its green permanence phenotype, the third oldest, sixth and ninth leaves of the ACSß / ACSß, acs2 / acs2 and acsß / acsß plants were subjected to darkness-induced senescence in 20 DAP by covering them with covers for 7 days. All the leaves were completely green at the beginning of the experiment and remained attached to the plants. The acsß / acsß plants were irrigated daily with water or 100 μM ACC for 7 days. After 7 days, the dark-induced senescence has started on the wild-type leaves (ie, ASCß / ASCß) although not progressing to the degree observed after a 2-week dark treatment. The degree of senescence induced by darkness increased as a function of the age of the leaf such that leaf 3 exhibited more senescence than leaf 6 or leaf 9 (which were younger), suggesting that competition for senescence increases with the age of the leaf. Dark-induced senescence was also observed in leaf 3 of the homozygous acs2 mutant although it was less pronounced by that observed in the corresponding wild-type leaves. Although none of the homozygous acsß leaves exhibited dark induced senescence consistent with the observations made in Fig. 4, dark-induced senescence similar to that of wild-type leaves was observed when the acsß leaves were irrigated with 100 μM ACC. for 7 days. The ACC treatment did not affect the acsß leaves that were not covered, demonstrating that the senescence observed for the covered acsß sheets was specific to the dark treatment. The determination of the level of chlorophyll a + b of sheet 3 confirmed the visual results in the leaves acsß stopped substantially more chlorophyll after the 7-day dark treatment than wild-type leaves but not when irrigating ACC 100 μm (Figure 20 Panel A). ACC treatment alone did not induce the premature loss of chlorophyll since chlorophyll was not lost from uncovered leaves of acsß mutants irrigated with ACC. The acs2 leaves stopped only a moderately larger amount of chlorophyll than the wild type leaves. Similar results were observed for leaf 6 and leaf 9 although the level of chlorophyll in these younger leaves was higher than in the older leaf 3 samples as expected (Figure 20 Panel A). Similar trends were observed for the total soluble leaf protein: the acsß leaves retained substantially more protein after dark treatment than the wild type leaves but not when irrigating ACC 100 μm (Figure 20 Panel B). Western analysis for ribulose biscarboxylase (Rubisco) showed a substantial loss of Rubisco from leaves b73 treated with darkness that was larger with older leaves (leaf 3) than younger leaves (leaf 9) (Figure 20 Panel C). The dark-treated acsß leaves retained substantially more Rubisco than the dark-treated wild-type leaves treated with dark and the acs2 leaves retained a moderate level of Rubisco (Figure 20 Panel C). Acsß sheets treated with darkness irrigated with ACC 100 μM they lost a quantity of Rubisco similar to that of leaves B73 treated with darkness suggesting that ACC supplemented the loss of expression ACC synthase. No loss of Rubisco was observed in the ACCESS leaves treated with ACC when they remained in the lumen demonstrating that ACC treatment alone did not reduce the Rubisco level. These data demonstrate that the green permanence phenotype, which implies the retention of chlorophyll and leaf protein such as Rubisco, can be complemented by exogenous ACC, suggesting that the delay in the senescence in these plants is a consequence of the reduction in the loss of ACC synthase expression in the acsß mutant. The reduction of ethylene delays the senescence of the natural leaf and reduces the loss of chlorophyll and protein drought is known to induce the premature onset of leaf senescence. To investigate whether the drought response is mediated by ethylene and to determine whether the reduction of ethylene emission can increase drought tolerance in corn, homozygous acsß and acsß mutant plants and wild type plants were grown in the field and under well-irrigated conditions (eight hours twice a week) and stressed water conditions (four hours per week for a period of one month that started approximately one week before pollination and continued for 3 weeks after the pollination). During the period of limited water availability, plants stressed by drought exhibited wilting and leaf curl, visible confirmation of stress. After the stress treatment with water,. the degree of senescence of the leaf and the function were measured. The senescence of the oldest leaves was evident in the wild type plants under well-irrigated conditions and even more significantly during the drought conditions. Similar results were observed for acs2 plants. In contrast, no visible sign of senescence was observed in the acsß under well-irrigated or drought conditions. Interestingly, the production of anthocyanin was also reduced in the acsß sheets. To confirm that the green permanence phenotype is related to increased levels of chlorophyll, the level of chlorophyll a and b was measured. Chlorophyll decreased with leaf age as well as with the age of the plants (Figure 17 Panels A and D). As expected, the largest decrease in chlorophyll correlated with the visible onset of old age. Under well-irrigated conditions, the level of chlorophyll in the acsß leaves (ACSd 0/0) was up to 8 times higher than the corresponding leaves of wild-type plants that had senescence initiated. Surprisingly, the level of chlorophyll in all acsß leaves, including young, substantially higher than wild-type plants (Figure 17 Panel A). The level of chlorophyll in the acs2 leaves (ACS2 0/0) was moderately higher than in the wild type plants. These results indicate that the increase in the chlorophyll content inversely correlates with the level of ethylene production: the moderate reduction in ethylene in the acs2 plants correlated with a moderate increase in the chlorophyll content while the large reduction in ethylene in the acsß plants it was correlated with a substantial increase in the chlorophyll content. These results also show that the reduction of ethylene increases the level of chlorophyll even in young leaves that are exhibiting the maximum leaf function (see below). Under drought conditions, the chlorophyll level was reduced in the mutant and wild-type plants but decreased to an even greater extent in the wild-type plants (Figure 17 Panels B-C). For example, the level of chlorophyll in leaf 5 of wild-type plants stressed with water decreased 2.5 times relative to non-drought plants while it decreased by only 20% in leaf 5 of acsß plants stressed with water ( Figure 17 Panel C). Consequently, the reduction of the ethylene emission resulted in a chlorophyll level in the older leaves of the acsß plants that was above 20 timeshigher than in the corresponding leaves of the wild type plants. As observed for non-drought plants, the chlorophyll level was highest in all acsß leaves, including the youngest ones. The chlorophyll content in the acs2 leaves also remained moderately higher under drought conditions than in the wild-type plants. Thus, the loss of the ACSß expression reduced the water stress responsiveness in that the chlorophyll content was substantially maintained under those stress conditions that have induced a significant loss of chlorophyll in the wild-type plants. The leaf protein also declined with the age of the leaf and with the age of the plant (Figure 18 Panel D). As observed for chlorophyll, the most substantial decrease in the protein correlated with the visible onset of senescence (Figure 18 Panel D). Under non-drought conditions, the protein level in the acsß leaves was up to 2 times higher than the corresponding leaves of wild-type plants that have started senescence (Figure 18 Panel A). As observed for chlorophyll, the protein level in all acsß leaves, including the youngest, was substantially higher than in wild-type plants and the level of protein in the acs2 leaves was moderately higher than in plants of wild type (Figure 18 Panel A). Exposure to the conditions of drought resulted in a larger decrease in protein in the older wild-type leaves that was observed in the treated leaves "(Figure 18 Panels BC) As was observed for non-drought plants, the protein level was higher In all acsß sheets, including the youngest ones, these results parallel those for chlorophyll indicate that the protein content inversely correlates with the level of ethylene emission.They also demonstrate that the loss of ACC synthase expression in the acsß mutant reduced the water stress responsiveness in that protein levels were substantially maintained under those stress conditions that have induced a significant reduction of proteins in wild type plants.The Ethylene Reduction Maintains the Function of the Leaf During Well Irrigated and Drought Conditions The maintenance of chlorophyll and protein on the acsß leaves suggests that the function of the leaf, for example, the ability to transpire and assimilate CO, can also be maintained. To investigate this, the proportion of transpiration, stomatal conductance and proportion of CO2 assimilation were measured on each leaf of acsß and wild type plants well irrigated at 40 DAP when the lower leaves of the wild type plants have reached senescence. The younger leaves of the acsß plants they exhibited a higher proportion of perspiration (Figure 5 Panel A) and stomatal conductance (Figure 5 Panel B) than the control plants, whereas no significant difference was observed in the older leaves. In contrast, the proportion of CO2 assimilation was substantially higher in all the leaves of the acsß plants than in the control plants (Figure 5 Panel C). Specifically, the older leaves of the acsß plants exhibited more than 2 times the highest proportion of assimilation of C02 than the wild type plants and the CO assimilation ratio; in younger leaves it was increased from 50 to 100% (Figure 5 Panel C). The effect of ethylene reduction on maintenance of leaf function under drought conditions was also investigated. The proportion of transpiration (Figure 5 Panel D) and stomatal conductance (Figure 5 Panel E) were significantly reduced in wild type leaves when subjected to drought conditions (ie, four hours per week during a month period starting about one week before pollination and continued until three weeks after pollination) while they remained largely unaffected on the acsß leaves, resulting in a substantially higher proportion of transpiration (Figure 5 Panel D) and increased stomatal conductance ( Figure 5 Panel E) by the mutant. In addition, the drought treatment result in a significant decrease in the assimilation rate of C02 in the wild type leaves but not in the acsß leaves, resulting in a 2.5 fold increase in the assimilation of C02 in the younger acsß leaves and up to an increase of 6 times in the older acsß sheets than in the control (Figure 5 Panel F). These results indicate that ethylene controls leaf function during drought conditions and a reduction in its production results in a delay in the leaf's aging in older leaves while maintaining leaf function in all leaves. thus providing greater tolerance to drought. Similar results, although less pronounced, were obtained for ACS2 (Figure 6 Panels A-C). DISCUSSION In summary, the ACC synthase mutants that affect the first stage in ethylene biosynthesis were isolated in corn. These mutants exhibited a delay in senescence, of the natural leaf, induced by darkness and induced by drought and a phenotype of green permanence. The delay in senescence was reversible after exposure to ethylene. The mutant leaves of. ACC synthase exhibited a proportion substantially higher assimilation of • C0_ during growth under normal drought conditions. Surprisingly, the improved leaf function was observed in all ACC synthase mutant leaves, including the youngest ones that have not entered the senescence programs either natural or drought-induced. These observations suggest that ethylene mediates maize's response to water stress and that the decrease in ethylene production serves as a means to maintain leaf performance during water stress and thus increase their tolerance to water conditions. of drought. As mentioned, ACC synthase mutants may have other advantageous phenotypes, for example, male sterility phenotypes, stacking resistance phenotypes, resistance to altered pathogens and the like. The above examples show that ethylene plays a significant role in regulating the onset of leaf senescence in maize either during growth under well-irrigated conditions or during drought conditions that normally induce premature leaf senescence. The reduction in ethylene emission that results from the loss of ACSß expression is largely responsible for directing the senescence of the natural leaf and induced by drought. While not intended to be limited by any particular theory, the loss of ACSß expression may directly delay entry into the senescence program or may affect the expression of ACC synthase of all gene members. The inactivation of ACS2 only reduced ethylene production by approximately 40% and resulted in a small increase in chlorophyll and protein. In contrast, the ethylene production in the acsß sheets was reduced to 90% and the acsß sheets contained substantially higher levels of chlorophyll and protein. These observations suggest that entry into the senescence program can be controlled by more than one member of the gene family. The level of chlorophyll and protein in the wild-type leaves was substantially reduced after water stress but remained unaffected in the acsß leaves. These results indicate two functions for ethylene in corn leaves: under normal growth conditions, ethylene can help maintain the correct level of chlorophyll and protein in a leaf, while during water stress, ethylene can serve to reduce the level of both. The observation that a 40% reduction in ethylene resulted in a moderate increase in chlorophyll and protein while a reduction of 90% resulted in a substantially larger increase in chlorophyll and protein suggests that these components of the Leaf can be quantitatively controlled by the level of ethylene produced in the leaves. Larger increases in chlorophyll and leaf protein could be expected if the production of ethylene were further reduced.

The loss of chlorophyll and protein in wild-type maize subjected to drought conditions was performed by decreased proportions of transpiration, stomatal conductance and assimilation of C0_. In contrast, maintenance of chlorophyll and protein levels in the leaves of acsß plants subjected to drought conditions was accompanied by maintenance of perspiration, stomatal conductance and CO uptake; These results suggest that ethylene reduction not only confirms a green permanence phenotype but actually maintains leaf function under stress conditions. The observation that ethylene controls the onset of leaf senescence is consistent with the function of this hormone in other species such as Arabidopsis and tomato (Davis and Grierson (1989) Identification of CDNA clones for tomato (Lycopersicon esculentum Mili.) MRNAs that accumulate during fruit ripening and leaf senescence in response to ethylene, Plant 179: 73-80, 7Abeles et al., (1992), Ethylene in Plant Biology (San Diego: Academic Press), Picton et al., (1993). teredfruit ripening and leaf senescence in tomatoes expressing an antisense ethylene-forming enzyme transgene Plant J. 3: 469-481; Grbic and Bleecker (1995) Ethylene regulates the timing of leaf senescence in Arabidopsis Plant J. 8: 95-102; John et al., (1995) Delayed leaf senescence in ethylene-deficient ACC-oxidase by (indicates (OsiACS ...) and japonica (OsjACS ...)), sequences of wheat are indicated by (TaACS ...) and banana sequences are indicated by (MaACS ...). In the analysis, the indicated ACC cassettes fall into two subfamilies. One of the subfamilies is further subdivided into ACC genes of monocotyledons (Zm (corn), Osi, Osj, Ta, Ma) and 'ACS genes of dicotyledons (At, Le). Several peptide consensus sequence alignments of ACC synthase sequences described herein, for example, (A47 (also known as ACS2 or ACC2 herein), A50 (also known as ACS7 or ACC7 herein), A65 ( also known as ACSd or ACC6 in the present)) of corn (Zm), with ACC synthase sequences from other species are shown in Figures 8-16. A Pret program is used (for example, available on the SeqWeb network (GCG) page) to determine the consensus sequence with different severities (for example, more severity (identical), severe (similar amino acids) or less severe (slightly similar amino acids). The severity is indicated in each figure after the "consensus sequence". The Space Weight is 8 and the Space Length Weight is 2. EXAMPLE 3: ACC ACTIVATES SYNTHASA BY EXPRESSION OF FORK RNA As previously mentioned, the cells of inactive plants and plants can be produced, by antisense tomato plants: molecular and physiological analysis Plant J. 7: 483-490). The observation that a reduction in the emission of ethylene would increase the level of chlorophyll and protein and increase the rate of assimilation of CO; On all the sheets, including the youngest ones, it was unexpected. This suggests that ethylene plays an active role in controlling aspects of leaf function before a leaf enters a senescence program. Equally unexpected was the observation that a reduction in ethylene will affect the water stress response of all leaves. These findings suggest that increased tolerance to drought conditions can easily be introduced into corn, and optionally other grain species, through a reduction in the level of ethylene produced in the leaves. EXAMPLE 2: SEQUENCE ALIGNMENTS AND PHYLOGENETIC ANALYSIS A phylogenetic analysis of the ACC synthase sequences described herein, for example, (A47 (also known as ACS2 or ACC2 herein), A50 (also known as ACS7 or ACC7 in the present), A65 (also known as ACSd or ACCd herein)) of maize, with ACC synthase sequences from other species, is shown in Figure 7 (ACSgrowtree2), where Arabidopsis sequences are indicated by (AtACS ... ), tomato sequences are indicated by (LeACS ...), rice sequences are indicated example, by introducing an ACC synthase polynucleotide sequence configured for RNA silencing or interference. This example describes cassette RNA expression cassettes for modifying ethylene production and the green permanence phenotype, for example, in corn. As previously mentioned, the inactivation of ACC synthase (s), for example, by the expression of hpRNA, can result in plant or plant cells having reduced expression (up to and including no detectable expression) of one or more ACC synthases . The expression of hairpin RNA molecules (hpRNA) specific for ACC synthase genes (eg, promoters, other untranslated regions or coding regions) that encode ACC synthase in plants can alter the ethylene production and the green permanence potential, sterility, resistance to stacking, etc. of plants, for example, through interference and / or RNA silencing. The hpRNA constructs of ACS2 (PHP20600) and ACS6 (PHP20323) were generated by linking a ubiquitin promoter to an inverted repeat of a portion of the coding sequence of either the ACS2 or ACS6 gene (See Figures 21 and 22, Panels A-C). Each construction was transformed into corn using the techniques of transformation mediated by Agrobacterium. The nucleic acid molecules and methods for preparing the constructions and transforming the corn were as previously described and known in the art; see, for example, sections in the present entitled "Vectors, Promoters and Expression Systems", "Plant Transformation", "Other Nucleic Acid Tests Proteins "and the following example" Corn Transformation. "The expression of specific hpRNA for either the ACS2 or ACS6 coding sequences resulted in maize plants that exhibited abnormalities in vegetative and reproductive growth, a total of 36 and 40 evidences Individual maize transgenic cultures were generated for the ACS2- and ACS6-hairpin constructs, respectively (Figure 23, Panels A and B) Approximately 10 evidence of low copy number per hpRNA construct were selected for further backcrossing and evaluation of the transgen.

The potential phenotype of green permanence for cross-lines subsequently comprising the transgene (s) of hpKNA, for example, as described herein (e.g., by visual inspection, photosynthetic activity measurements, content determination chlorophyll or protein or the like, under normal conditions and drought or other stress). EXAMPLE 4: TRANSFORMATION OF BIOLOGICAL CORN The inventive polynucleotides contained within a vector are transformed into embryogenic corn cs by bombardment of particles, gener as described by Tomes, D. et al., IN: Plant Cell, Tissue and Organ Culture: Fundamental Methods, Eds. 0. L. Gamborg and G. C. Phillips, Chapter 8, pages 197-213 (1995) and as summarized briefly below. Transgenic maize plants are produced by bombarding embryo-responsive immature embryos with tungsten particles associated with DNA plasmids. Plasmids typic comprise or consist of a selectable marker and an unselected structural gene, or a selectable marker and a polynucleotide sequence or subsequence of ACC synthase, or the like.

Particle Preparation: Fifteen mg of tungsten particles (General Electric), 0.5 to 1.8 μ, preferably 1 to 1.8 μ, and more preferably 1 μ, are added to 2 ml of concentrated nitric acid. This suspension is sonicated at 0 ° C for 20 minutes (Branson Sonifier Model 450, 40% output, constant duty cycle). The tungsten particles are formed into pellets by centrifugation at 10000 rpm (Biofuge) for one minute, and the supernatant is removed. Two milliliters of sterile distilled water are added to the pellet and brief sonication is used to resuspend the particles. The suspension is formed into pellets, one milliliter of pure ethanol is added from the pellet and the brief sonication is used to resuspend the particles. The rinsing and the pellet formation and resuspension of the particles is done twice more with sterile distilled water, and finally the particles are resuspended in two milliliters of sterile distilled water. The particles are subdivided into aliquots of 250 μl and stored in the frozen state. Preparation of the Particle-DNA Association of Plasmid: The extract of tungsten particles are sonicated briefly in a water bath sonicator (Branson Sonifier Model 450, 20% output, constant duty cycle) and 50 μl is transferred to a microcentrifuge tube. The vectors are typically cis: that is, the selectable marker and the gene (or other polynucleotide sequence) of interest are in the same plasmid. The plasmid DNA is added to the particles for a final DNA amount of 0.1 to 10 μg in 10 μL of total volume, and briefly sonicated. Preferably, 10 μg (1 μg / μL in TE buffer) of total DNA is used to mix the DNA and particles for bombardment.

Fifty microliters (50 μL) of sterile aqueous 2.5 M CaCl2 are added, and the mixture is briefly non-identical and rotated in a vortex. Twenty microliters (20 μL) of sterile aqueous 0.9 M spermidine are added and the mixture is briefly sonicated rotated in a vortex. The mixture is incubated at room temperature for 20 minutes with intermittent brief sonication. The particle suspension is centrifuged and the supernatant is removed. Two hundred and fifty microliters (250 μL) of pure ethanol are added to the pellet, followed by brief sonication. The suspension is formed in polotillas, the supernatant is removed and 60 μl of pure ethanol are added. The suspension is sonicated briefly before loading the particle-DNA agglomeration into macrocarriers. Tissue Preparation Immature maize embryos of variety Type II High are the target for transformation mediated by particle bombardment. This genotype is the Fl of two purebred, parental lines A and B, derived from the crossing of two known inbred maize lines, Al88 and B73. Both parents are selected for high competition from somatic embryogenesis, according to Ar strong et al., Maize Genetics Coop. News 65:92 (1991). Cobs from the Fl plants are autocrossed and the embryos are aseptically dissected from the developing caryopsis when the scutellum first becomes opaque. This stage occurs approximately 9-13 days after the pollination and more generally about 10 days of pollination, depending on growth conditions. The embryos are approximately 0.75 to 1.5 millimeters long. The ears are sterilized on the surface with Chlorides at 20-50% for 30 minutes, followed by three languages with sterile distilled water. The immature embryos are cultivated with the scepter oriented upwards, in the embryogenic induction medium comprised of basal N6 salts, Eriksson vitamins, 0.5 mg / 1 thiamine HCl, 30 gm / 1 saccharose, 2.88 gm / 1 L-proline , 1 mg / 1 of 2,4-dichlorophenoxyacetic acid, 2 gm / 1 Gelrite and 8.5 mg / 1 of AgN0. Chu and collaborators, Sci. Sin. 18: 659 (1975); Eriksson, Physiol. Plant 18: 976 (1965). The medium is sterilized by autoclaving at 121 ° C for 15 minutes and supplied in Petri dishes of 100 X 25 mm. AgN0 is sterilized in the filter and added to the medium after autoclaving. The tissues are cultured in complete darkness at 28 ° C. After about 3 to 7 days, more usually about 4 days, the embryo scutellum swells to about twice its original size and the protuberances on the coleorheal surface of the scutellum indicate the onset of embryogenic tissue. Up to 100% of embryos exhibit this response, but more commonly, the frequency of embryogenic response is approximately 80%.

When the embryogenic response is observed, the embryos are transferred to a medium comprised of modified induction medium to contain 120 gm / 1 of sucrose. The embryos are oriented with the coleorizal, the embryogenically responsive tissue, upwards of the culture medium. Ten embryos per Petri dish are located in the center of a Petri dish in an area approximately 2 cm in diameter. The embryos are maintained in this medium for 3-16 hours, preferably 4 hours, in complete darkness at 28 ° C just before bombardment with particles associated with plasmid DNA containing the selectable and non-selectable marker genes. To effect particle bombardment of the embryos, particle-DNA agglomerates are accelerated using a DuPont PDS-1000 particle acceleration device. The particle-DNA agglomeration is briefly sonicated and 10 μl is deposited on macrocarriers and the ethanol is allowed to evaporate. The macrocarrier is accelerated on a stainless steel screening screen by rupturing a polymer diaphragm (rupture disk). The rupture is effected by the pressurized helium. The speed of particle-DNA acceleration is determined based on the breaking pressure of the rupture disk. Expressions of the rupture disc from 200 to 1800 psi are used, with 650 to 1100 psi being preferred, and approximately 900 psi which is more highly preferred. Multiple discs are used to effect a range of rupture pressures. The support containing the embryo plate is placed 5.1 cm below the bottom of the macrocarrier platform (support # 3). To effect the bombardment of cultured immature embryo particles, a rupture disk and a macrocarrier with dry particle-DNA agglomerates with installed in the device. The pressure of He is supplied to the device and is adjusted to 200 psi above the breaking pressure of the rupture disk. A Petri dish with target embryos is placed in the vacuum chamber and located in the projected path of accelerated particles. A vacuum is created in the chamber, preferably approximately 28 pg Hg. After the operation of the device, the vacuum is released and the Petri dish is removed. The bombarded embryos remain in the osmotically adjusted medium during the bombardment, and 1 to 4 days subsequently. The embryos are transferred to the selection medium comprised of basal N6 salts, Eriksson vitamins, 0.5 mg / 1 thiamine HCl, 30 gm / 1 sucrose, 1 mg / 1 2,4-dichlorophenoxyacetic acid, 2 gm / 1 Gelrite, 0.85 mg / 1 of Ag N03 and 3 mg / 1 of bialaphos (Herbiace, Meiji). The bialafos is added to stylize the filter. The Embryos are subcultured to the fresh selection medium at intervals of 10 to 14 days. After approximately 7 weeks, the embryogenic tissue, putatively transformed for both selectable and non-selectable marker genes, proliferates from approximately 7% of the bombarded embryos. The putative transgenic tissue is rescued, and that tissue derived from the individual embryos is considered to be an evidence and is independently propagated on the selection medium. Two cycles of clonal propagation are achieved by visual selection for the smallest contiguous fragments of the organized brionic tissue. A tissue sample of each evidence processed to recover the DNA. The DNA is restricted with a restriction endonuclease and probed with primer sequences designed to amplify DNA sequences overlapped to the ACC synthase and non-ACC synthase portions of the plasmid. The embryogenic tissue with amplifiable sequence is advanced for the regeneration of the plant. For the regeneration of transgenic plants, the embryonic tissue is subcultured to a medium comprising MS salts and vitamins (Murashige &Skoog, Physiol. Plant 15: 473 (1962)), 100 mg / 1 myo-inositol, 60 gm / 1 sucrose, 3 gm / 1 Gelrite, 0.5 mg / 1 zeatin, 1 mg / 1 indole-3-acetic acid, 26.4 ng / 1 cis-trans-abscisic acid and 3 mg / 1 of bialaphos in Petri dishes of 100 X 25 mm, and incubated in the dark at 28 ° C until the development of mature, well-formed somatic embryos can be observed. This requires approximately 14 days. The well-formed somatic embryos are opaque and cream colored, and are comprised of an identifiable scutelle and coleoptile. The embryos are individually subcultured to a germination medium comprising MS salts and vitamins, 100 mg / 1 myo-inositol, 40 gm / 1 sucrose and 1.5 gm / 1 Gelrite in 100 X 25 mm Petri dishes and incubated under a photoperiod of 16 hours of light: 8 hours of darkness and 40 meinsteinsnf ^ sec-1 of cold white fluorescent tubes. After approximately 7 days, the somatic embryos have germinated and produced a well-defined shoot and root. The individual plants are subcultured to the germination medium in 125 X 25 mm glass tubes to allow further development of the plant. The plants are kept under a photoperiod of 16 hours of light: 8 hours of darkness and 40 meinsteinsirSseg-1 of cold white fluorescent tubes. After approximately 7 days, the plants are well established and are transplanted to the horticultural soil, hardened and placed in pots mixed with commercial greenhouse soil and grown to sexual maturity in a greenhouse. An elite inbred line is used as a male part to pollinate regenerated transgenic plants.

MEDIATED BY AGROBACTERIUM When the transformation mediated by Agrobacterium is used, the Zhao method is employed as in the PCT patent publication W098 / 32326, the contents of which are incorporated herein by reference. Briefly, immature embryos are isolated from maize and embryos contacted with a suspension of Agrobacterium (stage 1: the infection stage). At this stage the immature embryos are preferably immersed in a suspension of Agrobacterium for the initiation of the inoculation. The embryos are co-cultivated for a time with the Agrobacterium (stage 2: the co-culture stage). Preferably the immature embryos are cultured in solid medium after the infection stage. After this period of co-cultivation an optional "resting" stage contemplates. In this resting stage, the embryos are incubated in the presence of at least one known antibiotic that inhibits the growth of Agrobacterium without the addition of a selective agent for plant transformants (step 3: resting step). Preferably the immature embryos are cultured on solid media with antibiotic, but without a selection agent, for the elimination of Agrobacterium and for a resting phase for the infected cells. Next, the inoculated embryos are cultured in the medium containing a selective agent and the callus transformed into growth is recovered (stage 4: the selection stage). Preferably, immature embryos they are grown on solid medium with a selective agent resulting from the selective growth of transformed cells. The callus is then regenerated in plants (stage 5: the regeneration stage) and preferably the calluses developed on the selective medium are cultivated in the solid medium to regenerate the plants. EXAMPLE 5. EXPRESSION OF TRANSOGENES IN MONOCOTILEDONAS A plasmid vector is constructed comprising a preferred promoter operably linked to an isolated polynucleotide comprising a polynucleotide sequence or subsequence of ACC synthase (eg, selected from SEQ ID NO: 1-6 and 10). This construction can then be introduced into corn cells by the following procedure. Immature corn embryos are dissected from developing caryopsis derived from crosses of corn lines. Embryos are isolated 10 to 11 days after pollination when they are 1.0 to 1.5 mm long. The embryos are then placed with the shaft side facing down and in contact with the N6 medium agarose sonicated (Chu et al., (1975) Sci. Sin. Peking 18: 659-668). The embryos are kept in the dark at 27 ° C. Reliable embryogenic callus, consisting of masses of cells undifferentiated with proembryoids and somatic embryoids carried in suspension structures, the scutellum proliferates of these immature embryos. The embryogenic callus isolated from the primary explant can be cultured in the N6 medium and subcultured in this medium every 2 to 3 weeks. Plasmid p35S / Ac (Hoechst Ag, Frankfurt, Germany) or equivalent can be used in transformation experiments in order to provide a selectable marker. This plasmid contains the Pat gene (see European Patent Publication 0 242 236) which encodes phosphinothricin acetyl transferase (PAT). The PAT enzyme confers resistance to herbicidal glutamine synthetase inhibitors such as phosphinothricin. The p35S / Ac gene pat is under the control of the 35S promoter of the Cauliflower Mosaic Virus (Odell et al., (1985) Nature 313: 810-812) and comprises the 3 'region of nopaline synthase from T-DNA of the Ti plasmid of Agrobacterium tumefaciens. The particle bombardment method (Klein et al., (1987) Nature 327: 70-73) can be used to transfer genes to callus culture cells. According to this method, gold particles (1 μm in diameter) are coated with D? A using the following technique. Ten μg of D? A of plasmid are added to 50 μL of a suspension of gold particles (60 mg per mL). Calcium chloride (50 μL of a 2.5 M solution) and spermidine free base (20 μL of a 1.0 M solution) are added to the particles. The suspension is rotated in a vortex during the addition of these solutions. After 10 minutes, the tubes are centrifuged briefly (5 sec at 15,000 rpm) and the supernatant is removed. The particles are resuspended to 200 μl of pure ethanol, centrifuged again and the supernatant is removed. The ethanol rinsing is performed again and the particles resuspended in a final volume of 30 μL of ethanol. An aliquot (5 μL) of the particles of another coated with DNA can be placed in the center of the Kapton flywheel (Bio-Rad Labs). The particles are then accelerated in the corn tissue with a Biolistic PDS-1000 / He (Bio-Rad Instruments Hercules CA), using a helium pressure of 1000 psi, a space distance of 0.5 cm and a flying distance of 1.0 cm. For the bombardment, the embryogenic tissue is placed on filter paper on the N6 medium solidified in agarose. The tissue is arranged as a thin layer and covers a circular area approximately 5 cm in diameter. The petri-box containing the tissue can be placed in the camera of the PDS-1000 / He approximately 8 cm from the screening screen. The air in the chamber is then evacuated to a vacuum of 28 inches Hg. The macrocarrier is accelerated with a helium shock sling using a rupture membrane that it explodes when the He pressure in the shock tube reaches 1000 psi. Seven days after the bombardment the tissue can be transferred to the N6 medium containing glufosinate (2 mg per liter) and lacks casein or proline. The tissue continues to grow slowly on this medium. After an additional 2 weeks the tissue can be transferred to fresh N6 medium containing glufosinate. After 6 weeks, areas of approximately 1 cm in diameter of callus actively growing can be identified some of the plates containing medium supplemented with glufosinate. These calluses can continue to grow when subcultured in the selective medium. Plants can be regenerated from the transgenic callus by first transferring tissue clusters to N6 medium supplemented with 0.2 mg per liter of 2,4-D. After two weeks the tissue can be transferred to the regeneration medium (Fromm et al., (1990) Bio / Technology 8: 833-839). EXAMPLE 6. EXPRESSION OF TRANSGENES IN DICOTILEDONAS Soy embryos are bombarded with a plasmid comprising a preferred promoter operably linked to a heterologous nucleotide sequence comprising a polynucleotide sequence or subsequence of ACC synthase (eg, selected from SEQ ID NO: l-6 and 10), as follow. To induce somatic embryos, cotyledons of 3-5 mm in length are dissected from immature seeds sterilized on the surface of the soybean variety A2872, then grown in light or dark at 26 ° C on an appropriate agar medium for six days. to ten weeks. Somatic embryos that produce secondary embryos are then excised and placed in a suitable liquid medium. After repeated selection for clusters of somatic embryos that multiply as embryonic globular stage, early, suspensions are maintained as described below. Soy embryogenic suspension cultures can be maintained in 35 ml of liquid medium on a rotary shaker, 150 rpm, at 26 ° C with fluorescent lights in a program of 16: 8 hours day / night. The cultures are subcultured every two weeks by inoculating approximately 35 mg of tissue in 35 ml of liquid medium. Soybean embryogenic suspension cultures can then be transformed by the particle gun bombardment method (Klein et al. (1987) Nature (London) 327: 70-73, North American patent No. 4,945,050). A DuPont Biolistic PDS1000 / HE instrument (helium retro-fit) can be used for these transformations. A selectable marker gene that can be used to facilitate the transformation of soybean is a transgene composed of the 35S promoter of the Cauliflower Mosaic Virus (Odell et al., (1985) Nature 313: 810-812), the hygromycin phosphotransferase gene of the plasmid pJR225 (from E. coli; collaborators, (1983) Gen 25: 179-188), the 3 'region of the nopaline synthase gene of TD? A of the Ti plasmid of Agrobacteri um tumefaciens. The expression cassette of interest, comprising the preferred promoter and a heterologous ACC synthase polynucleotide, can be isolated as a restriction fragment. This fragment can then be inserted into a restriction site unique to the vector carrying the marker gene. To 50 μl of a suspension of gold particles of 1 μm of 60 mg / ml is added (in order): 5 μl of D? A (1 μg / μl), 20 μl of spermidine (0.1 M) and 50 μl of CaCl; (2.5 M). The particle preparation is then stirred for three minutes, rotated in a microcentrifuge for 10 seconds and the supernatant is removed. The particles coated with D? A are then washed once in 400 μl of 70% ethanol and resuspended in 40 μl of anhydrous ethanol. The D? A / particle suspension can be sonicated three times for one second each time. Five microliters of gold particles coated with D? A are then loaded with a macrocarrier disk. Approximately 300-400 mg of a two-week-old suspension culture is placed in a box empty 60x15 mm petri and the residual liquid removed from the tissue with a pipette. For each transformation experiment, approximately 5-10 tissue plates are normally bombarded. The rupture pressure of the membrane is adjusted to 1100 psi and the chamber is evacuated to a vacuum of 28 inches of mercury. The fabric is placed approximately 3.5 inches away from the retention screen and bombarded three times. After bombardment, the tissue can be divided in half and placed back in liquid and cultivated as described above. Five to seven days after the bombardment, the liquid medium can be exchanged with fresh medium, and eleven to twelve days after the bombardment with fresh medium containing 50 mg / ml hygromycin. This selective medium can be renewed every week. Seven or eight weeks after the bombardment, the transformed, green tissue can be observed growing from necrotic, non-transformed embryogenic clusters. The isolated green tissue is removed and inoculated into individual flasks to generate new, cloned, propagated transformed embryogenic suspension cultures. Each new line can be treated as evidence of independent transformation. These suspensions can then be subcultured and maintained as groups of immature or regenerated embryos in whole plants by maturation and germination of somatic embryos individual It is understood that the examples and embodiments described herein are for illustrative purposes only and that various modifications or changes in view of them will be suggested to those skilled in the art and will be included within the spirit and incubency of this application and the scope of the claims adtas. While the above invention has been described in some detail for purposes of clarity and understanding, it will be clear to a person skilled in the art to start from a reading of this description that various changes in form and detail can be made without departing from the actual scope of the invention. the invention. For example, all the techniques and apparatuses described in the above can be used in various combinations. All publications, patents, patent applications and / or other documents cited in this application are incorporated by reference in their entirety for all purposes to the same degree as if each individual publication, patent, patent application and / or other document were individually indicated that it is incorporated by reference for all purposes.

GACAACAAAT AATAGCCAAT TGTAGCCCCT CGCCATCTTT CCTTGTTTGG GTAACGTTTC AAAMTTAGG GGGTGTTTGG TTTCTAGGGA CTAATGTTTA GTCCCTTCAT TTTATTCCAT TTTAGTATAT AAATTGTCAA ATATAAAAAC CAAAATAGAG TTTTAGTTTC TATATTTGAC AATTTTAGAA CTAAAATGAA ATAAAATGTA GGGACTAAAG TATAAACTAA ACACCGCCTT ACCTCGATCA CGAACCTCTA AAAGTAAGTA GCACCCTCCT CCCCCACAGT CAAATCAACA TAATACAGTA CAATAGACCT TGTTAGTCGC ATGGATGATT GTCGTCAAGT GGGCAACGCA ATCTAGTCAC GTAAGGAAAA CCATGCACGT TGTTCATACA CGGTCTGTTT CCATGCGACT TTAATTTCCA CGCACGTTTG CATCGTTGAC CAACCAACTG AACGTGCCTG TAGGTCCCGC ACAGCAACGT AAGCATATGC ATGCACGTAC GACGTACGGC ACGGGAAAAA AATTCTGCAC ACCGTATTTT ACAGCTCTTC ATATCCACCA CATGTAGCGG CCCCACAAAA AACAGATTAA AATTTGCAAC TTAATCCTTA AGTAATTTGT TTTTCTTCTA TTTATATAGA TTATCAGTTG ATGGATGTGT GAAGTTGTAA AAGAGATTAT TTGTATCCAG GATTAAAATA ATTTTCCGTA CGGCACGCCT GCAGTACTCA TTCTCGCCAG CCCTGAGCCC CTGATATATG ACACGCTTTT CATTGTTCAC ACAGTTTCGA CCGTGACTGT TGCTGGAGGT CAGGCGTGAA GCTGCTGCCC ATCGAATGCC ACAGCTCAAA CAACTTCACC CTCACACGGG AGGCGCTCGT GTCGGCCTAC GACGGCGCGC GGAGGCAGGG 'CGTCCGCGTC AAGGGCGTCC TCATCACCAA CCCCTCCAAC CCGCTGGGCA CCACCATGGA CCGCGCCACG CTGGCGATGC TCGCCAGGTT CGCCACGGAG CACCGTGTCC ACCTCATCTG CGACGAGATC TACGCGGGCT CCGTCTTCGC CAAGCCGGAC TTCGTGAGCA TCGCCGAGGT CATCGAGCGC GACGTCCCGG GCTGCAACAG GGACCTCATC CACATCGCGT ACAGCCTCTC CAAGGACTTC GGCCTCCCGG GCTTCCGCGT CGGCATCGTC TACTCGTÁCA ACGACGACGT CGTGGCCTGC GCGCGCAAGA TGTCCAGCTT CGGCCTCGTC TGCTCGCAGA CGCAGCACTT CCTGGCGAAG ATGCTGTCGG ACGCGGAGTT CATGGCCCGC TTCCTCGCGG AGAGCGCGCG GCGGCTGGCG GCGCGCCACG ACCGCTTCGT CGCGGGACTC CGCGAGGTCG GCATCGCGTG CCTGCCCGGC AACGCGGGGC TCTTCTCGTG GATGGACCTG CGGGGCATGC TCCGGGACAA GACGCACGAC GCGGAGCTGG AGCTGTGGCG GGTCATCGTA CACAAGGTGA AGCTCAACGT GTCGCCCGGC ACGTCGTTCC ACTGCAACGA GCCCGGCTGG TTCCGCGTCT GCCACGCTAA CATGGACGAC GAGACCATGG AGGTCGCGCT CGACAGGATC CGCCGCTTCG TGCGCCAGCA CCAGCACAAG GCCAAGGCCG AGCGCTGGGC GGCCACGCGG CCCATGCGCC TCAGCTTGCC GCGCCGGGGA GGCGCCACCG CTTCGCACCT CCCCATCTCC AGCCCCATGG CGTTGCTGTC GCCGCAGTCC CCGATGGTTC ACGCCAGCTA GTCACCGAGC ATCCGGCAAG ACTGGCTGTA GGGTGTGCCC GTACATCCGT ACGTACACCT TTTTTTTCCCA TTCACGTGAC TGCAATCAAG TCTATGGGAT GGTTGACAAA AGACTATCTA 'GACAAGAGTG GGCGTAGTAC GTAACTAGTT TGACGTTGTA CAGGCGTCAG CAGGTATCGG TAAGCAGCTA GTCAAAAGCA CGCAAGCAGG ACGCATTTGT CCTCGATACT TTCGTGTAAA TCTCTCTCTA TTTTTTTTTG CGAAATTCGC GTGTATGGT TGTTTTGACG TTGGTATAAA GTATGGTAGA ATAACGATGG GAAATGGCAA TTTAGTCCTC CCGATCAATT GTTATTGTAA ACCACTGACG AAAGTTAAGA ACAGAAGCTG TACCAGAAGG GTGAATAAAA ATACCACATA GGTATTGAAT TAATAATCTA TGTATTTCGA GTTACTCCTG CAAGATATCT ATTTTTTTCAT GCTGTGCTGG CCACATTTGC CTCTTCTTCA AACTAGTTTC TCGCA SEQ ID NO: 2 Gen ACSD CGGCTAGTTT TGATAGTTAG ACGATGTTCT GACAGCGCAC CAGACAGTAA (ACC6) CCAGTGACAG TCCGGTGCCT GGCTAAATAT CGAGCCAGCG AACAGCGCGC TCTCGGGTTT CTACGGGGGC AGAGGGTTGC TCTCGGGGCA TTCTTGTGCT Sequence CACTGTCAGG GGGAGCACCA GACAGTCCGG TGCACAGCGA ACAGTCTGAT gene GCCCCTAGGT CAGCAAGTCA AAGTTCTCTt CCTTAGATTT TTCTAAACCG ? CC TTTTCGTTTT AACTTGTGAG TGAGTTATCG AGTGACACCT AGCACTAGTT synthase GTGAGTATGA ACACCAACAC TATATTAGAT TTCTCTTGGT CAAACTACTC (A65) ATCCACAACC ACTCTTTATA GTACGGCTAA AATAAAAATA GAAGTCCTAA from Zea CTTTATACCA AGTGTCAACA ACTCCTTCGG ACACTTAGAA TATAAAGTCC mayb TTCATCTTTT GTTTCGCCTT TTTCCGCCGT CGCTTCAAGT TCTCATCCGA (Zm) GGGATTGTTT TATCGTTGTA GTGCAACTTC ATGCAATGTG ACCTAACTTG Inbred CCATTTGCTC TTCAAAACAC ACGTTAGTCA TATAATATTA CGTTGTCATT < B73 '(= AATCTCTATC GATATTTTTC ACCCATTACG TTGTCACTAG ATGCTTTCAC fragment CCATTTCGAT TTCAGACGAT GTCTTCGGAC GTTGCGGGCC ATGTGTCCAA per ACC6 ACGTGGTTAA GTGTGGTCGG GAAATACCCG ATCGAGGTTG AGTTCGGCCT originalTCGCTCCGAC ACCCAGCCGT GTCATTACTG TCATATATAT TGTAGCAATG mind TCAAAAAAAA TCAAAACATT GAGTATGACG TATAGGGCAC ATATGTCATT isolated AAACTTATTC AGTGTAATGA TATATTATCA TCACGGGACT of endogámico TTTTTTTAAT GTATGTATTA GATTACCTCT GCCATGCACT ATACAAACAG CTACGCCGCA GTCGCAAGCA AACAGGCTCT AAAAGGCTTC AGTCGGAGAA GGATATGAGA * Oh43 ') GCGGTGAGTA CCAAACGGGT ATCTTCCCCT TCCAAATGAT ATAAGCCTAC TTGTTTGACC CCAGCCCGCA GGCAGTCATC TGCTATAATA GGCTAATACA ACTTGTGTAC TCTAGTCTGC TCTCGCCGCG TTGTCCGCAT GCTGAACCCG CGATGTTAAC ACCTCCCTGA ACGAGTCCTC TGTTCCTCAA CTGAAATTCA GCAATAAAAG GAAAAATCCG CGGTCCCTGT CCCTGTCCAG CACCGCACTC TCGCACTTGT GCTGCAGGCT TCTGAGCTCG GCACCTGCTG CTAGCTGCTG CTATATATAG ACGCGTTTTG GGGTCACCAA AACCACCAGC TGATCAACAG CTAGCTTCAT TCCTCTGCCT- "CTCTCTCCCT CCTTCGCCAA CTGGCCATCT CTGTTGTCTC TCGCTAGCTA GCTCGCTCGC TCGCTCGCCA GTCACCACAC ACACACACAC ACACTGTGTG TCTGTGCCTG ACGCCGCCCC CCAGTTTCAA ACGAACGACC CAGCCAGAAA CGCGCGCGCG CCAAAGCTAC GTGAGTGACG TGGCAGCATG GTGAGCATGA TCGCCGACGA GAAGCCGCAG CCGCAGCTGC TGTCCAAGAA GGCCGCCTGC AACAGCCACG GCCAGGACTC GTCCTACTTC CTGGGGTGGG AGGAGTATGA GAAAAACCCA TACGACCCCG TCGCCAACCC CGGCGGCATC ATCCAGATGG GCCTCGCCGA GAACCAGCTG TCCTTCGACC TGCTGGAGGC GTGGCTGGAG GCCAACCCGG ACGCGCTCGG CCTCCGCCGG GGAGGCGCCT CTGTATTCCG CGAGCTCGCG CTCTTCCAGG ACTACCACGG CATGCCGGCC TTCAAGAATG TGAGTGCCTG CTAGCTTACT CATTCCCAGG CAGGCAGGCA GCCAGCCACG GCATGCCGAA CCAGTCTGAC CTCTCTCGCG CACATGAATG CGTGATTCCC GCAGGCATTG GCGAGGTTCA TGTCGGAGCA ACGTGGGTAC CGGGTGACCT TCGACCCCAG CAACATCGTG CTCACCGCCG GAGCCACCTC GGCCAACGAG GCCCTCATGT TCTGCCTCGC CGACCACGGA GACGCCTTTC TCATCCCCAC GCCATACTAC CCA5GGTATG TGTGTGTGTT GCCTTGTACT TACTCGTCGC CGCAAGTACT TGCAGTAGGG AACGTGCAAG TGGCGGCGGG GCGGCGTCTG GGTGTCGCCG CGATGCACGT TACTGCTATT AAAGTAGTAG TAGTACACTA ATAGCTAGGC CCACCACAGC ACACGATGAC ATGACGAACG ATGGATGGGA ACGGCTGCTG ACTGGGCCTG CTTGCTCTTG TCTGCAGGTT CGACCGTGAC CTCAAGTGGC GCACCGGCGC GGAGATCGTC CCCGTGCACT GCACGAGCGG CAACGGCTTC CGGCTGACGC GCGCCGCGCT GGACGACGCG TACCGGCGCG CGCAGAAGCT GCGGCTGCGC GTCAAGGGCG TGCTCATCAC CAACCCTTCC AACCCGCTGG GCACCACGTC GCCGCGCGCC GACCTGGAGA TGCTGGTGGA CTTCGTGGCC GCCAAGGGCA TCCACCTGGT GAGCGACGAG ATATACTCGG GCACGGTCTT CGCGGACCCG GGCTTCGTGA GCGTCCTCGA GGTGGTGGCC GCGCGCGCCG CCACGGACGA CGGCGTCGTC GGCGTTGGGC CGCTGTCGGA CCGCGTGCAC GTGGTGTACA GCCTGTCCAA GGACCTGGGC CTCCCGGGGT TCCGCGTGGG CGCCATCTAC TCGTCCAACG CCGGCGTGGT CTCCGCGGCC ACCAAGATGT CGAGCTTCGG CCTGGTGTCG TCCCAGACGC AGCACCTCCT GGCGTCGCTC CTGGGCGACA GGGACTTCAC GCGGAGGTAC ATCGCGGAGA ACACGCGGCG GATCAGGGAG CGGCGCGAGC AGCTGGCGGA GGGCCTGGCG GCCGTGGGCA TCGAGTGCCT GGAGAGCAAC GCGGGGCTCT TCTGCTGGGT CAACATGCGG CGCCTGATGC GGAGCCGGTC GTTCGAGGGC GAGATGGAGC TGTGGAAGAA GGTGGTCTTC GAGGTGGGGC TCAACATCTC CCCGGGCTCC TCCTGCCACT GCCGGGAGCC CGGCTGGTTC CGCGTCTGCT TCGCCAACAT GTCCGCCAAG ACGCTCGACG TCGCGCTCCA GCGCCTGGGC GCCTTCGCGG AGGCCGCCAC CGCGGGGCGC CGCGTGCTTG CCCCCGCCAG GAGCATCAGC CTCCCGGTCC GCTTCAGCTG GGCTAACCGC CTCACCCCGG GCTCCGCCGC CGACCGGAAG GCCGAGCGGT AGCCGGTCCC CGTCCGCGCC GACCGCACGT GCTCAGCTCA GCAGCTTCAC AGCTCACCAC TGGGGCAGGT GAGGGGCGGC AAGGTGACGT TCGACCCCGA CCGCGTCGTC ATGTGCGGAG GAGCCACCGG CGCGCAGGAC ACTCTCGCCT TCTGCCTCGC TGACCCGGGC GACGCCTACC TCGTGCCGAC GCCTTATTAC CCAGCGTATG TTCTGACGTC ACCCTTGTAC TGCCAAACTA CTACTCAGGT CCTAGTCATA TCCGTAGACA CGAAAGGGTG GGTGGGTCTG GGTTGTTGGT TGGTCAAGAG CACGCAAAAT TGAGCTAATT CGACTACGTA CGTGTCAATG TCAACTAGCC ACTTATCTTT CCTTGTTTGG GTAAAGTTTC AAAACTTATT AACTCGATCA GGAACCTCTC TAAAAAGCAT TCACCTATTT TTCCCCCGTA AGGCGGTAAC CAAATCTAAA CGATATACCC TTGTTAGTCG CACTGATGAC TGCATTGTCG TCAAGTGGAC AACGCAATCT AGTCACGCGA CCTCTAAGGA AAACCACGCA CGTATACGCA CTTCGTGCAC GGTCTGTTCC ACGCGACTTT AGTTTCCATG CACGTTTACA TCGTTGACCA TCCGCAGTCC GCACAGCAAC GTAAGCATAA ACATGCACGC ACGACGTACG GCACACCGTA CCTGTTCCTC TCGAGGGCTG AGACCCTGAC ACGTTTTTTT CGTTGTGTGG TGATCACAGT TTCGACCGCG ACTGTTGCTG GAGGTCAGGA GTGAAGCTGC TGCCCATCGA ATGCCACAGC TCGAACAACT TCACCCTCAC CAGGGAGGCG CTCGTGTCGG CCTACGACGG CGCGCGGAGG CAGGGCGTCC GCGTCAGGGG CATGCTCATC ACCAACCCCT CCAACCCGCT GGGCACCACC ATGGACCGCG GCACGCTGGC GATGCTCGCC GCGTTCGCCA CAGAGCGCCG CGTCCACCTC ATCTGCGACG AGATCTACGC GGGCTCCGTC TTCGCCAAGC CGGGCTTCGT GAGCATCGCC GAGGTCATCG AGCGCGGCGA CGCCCCGGGC TGCAACAGGG ACCTCGTCCA CATCGCGTAC AGCCTCTCCA AGGACTTCGG CCTCCCGGGC TTCCGCGTCG GCATCGTCTA CTCCTACAAC GACGACGTGG TGGCCTGCGC GCGCAAGATG TCCAGCTTCG GCCTCGTCTC GTCGCAGACG CAGCACTTCC TGGCGATGAT GCTCGCCGAC GCGGAGTTCA TGGCACGCTT CCTCGCGGAG AGCGCGCGGC GGCTGGCGGC GCGCCACGAC CGCTTCGTCG CGGGCCTCCG CGAGGTCGGC ATCGCGTGCC TGCCGGGCAA CGCGGGCCTC TTCTCGTGGA TGGACCTGCG GGGCATGCTC CGGGAGAAGA CGCACGACGC GGAGCTCGAG CTGTGGCGGG TCATCGTACA CAGGGTGAAG CTCAACGTGT CGCCCGGCAC GTCGTTCCAC TGCAACGAGC CCGGCTGGTT CCGCGTCTGC TACGCCAACA TGGACGACGA CACCATGGAG GTCGCGCTCG ACCGGATCCG CCGCTTCGTG CGCCAGCACC AGCACAGCAA GGCCAAGGCC GAGCGCTGGG CGGCCACGCG GCCCCTTCGC .CTCAGCTTGC CGCGCCGGGG AGCAACCACC GCTTCGCATC TCGCCATCTC CAGCCCCTTG GCGTTGCTGT CGCCGCAGTC CCCGATGGTC CACGCCAGCT AGGTAGTCAC CGAGCGTTCG GTAAGACTGG CTGTAGGTTG TGCCCTCACA TGACTGCAAA CAAGTGGACA AAAAAAAAGA CAAGACTAAT AAAGGGCGTA CGTAGCTAGC TTGACATTAC ACAGAGTGAC AGAGACGTTG CACAGGCGTC AGCAGGCGTC GGCGGTAAGC AGCTAGTCAA GTAGGACGCA TTTGTCCTCG ATTTTTTCGT GTTTTTTTTT TGACGAAGGG GCGAAGCCCC CTATTTCATT AAGAAATAGG AAAGTATGAA ACAACCGCAC CCACGCGGTA GGACCTCCAA AAAGAACAGC CACGGCCAGA AAGTAATCTA GACTCTAAAC ACTATCGCTA GATCAGTGAA GAGACTÁTGÁ TAACAGGGAA AGTTTTGGCC TACGA AGÁGC TACATAAGAC TTTCTTATAT ACAACCAACC AAGACAGGCA GAAGCCACAA AAGACCTGAA CAGAATGGCC AACAAAAGAC AGACAACTAT CCCAACAAGG TTTCACAGCT TCAGCATCTT TGTCATCCAG AAATCCGCCT GTCAAGAGGA CACCACCCCA AGGCCCTCCC GAAAGCTTCA CTTGCCGTCT TTCGGATTAA CCTGCTTCCT AGCACCACCA TTCTTTGCTC CTTCTTTTTC TGACGAATCG CCCAAGAATC CAACCAGAAG CAGCAAAGAA AAATGATGTT AGATGGGTCA AGTAAATGAC TATTCCCAAA ACACCAATCA TTCCTAGTGC GCCAAATAGC CCAGAATAAA GCACCACAAC CAAATAACAC CAACTGAGCC ATCGTGTCTT TTGGTTTACA AAACCAATTG TCATACAAAT CTTTGATATT TTTTGGAATA GATCTCAAAT TCAGGGCCAC TTGAATAACT CTCCACATGT ATTGAGCAAT GGGGCAATAG AAAAA SEQE > 4 cDNA ATGSCCGGTGGTAGCAGTGCCGAGCAGCOX.CTATCC-AGGATCGCCTCCGGCGATGIGCCAC ACS2 GGCGAGAACTCGTCCTACTTCGACGGGTGGAAGGCCTACGACATGGACCCTTTCGACCTG (ACC2) CGCC ^ CAACCGCGACGGCGTCATCCAGATGGGCCTCGCCGAGAACCAACTGTCCCTßGAC CTGA-CGAGCAATGsA_CA-GGAGCACCCGGA__CGTC < ^ TCTGCACGGCGCAGGGAGCG TCG_ \ GOWCAGGAGGATAGCCAACOTCCA__ACTACIACGGCC (-CCGGAGTT (_AGA_AG ACC tíCGATGGCC ^? GTTCA-GGGCCAGGTGAGGGC8GGAAGGW-ACG-TCGACCCCGACCGC

Claims

CLAIMS 1. An isolated or recombinant inactive plant cell, characterized in that it comprises at least one interruption in at least one endogenous ACC synthase gene, wherein the interruption inhibits the expression or activity of at least one ACC synthase protein compared to a corresponding control plant cell lacking interruption, wherein the at least one endogenous ACC synthase gene comprises a nucleic acid sequence, or complement thereof, comprising at least about 70% sequence identity to the SEQ ID NO: 1 (gACS2), SEQ ID NO: 2 (gACSβ) or SEQ ID NO: 3 (gACS7).
2. The plant cell according to claim 1, characterized in that the at least one endogenous ACC synthase gene comprises a nucleic acid sequence, or complement thereof, comprising at least about 85% sequence identity to the SEQ ID NO: 1 (gACS2), SEQ ID NO: 2 (gACSß) or SEQ ID NO: 3 (gACS7).
3. The plant cell according to claim 1, characterized in that the at least one gene Endogenous synthase ACC comprises two or more endogenous ACC synthase genes.
4. The plant cell according to claim 1, characterized in that the at least one gene Endogenous synthase ACC comprises three or more endogenous ACC synthase genes.
The plant cell according to claim 1, characterized in that the at least one interruption results in the production of reduced ethylene by the inactive plant cell as compared to the control plant cell.
6. The plant cell according to claim 1, characterized in that the at least one interruption comprises one or more transposons, wherein the one or more transposons are in the at least one endogenous ACC synthase gene.
7. The plant cell according to claim 6, characterized in that the at least one interruption is a homozygous interruption in the at least one ACC synthase gene.
8. The plant cell according to claim 6, characterized in that the at least one interruption is a heterozygous interruption in the at least one ACC synthase gene.
9. The plant cell according to claim 1, characterized in that the at least one interruption comprises one or more point mutations, wherein the one or more point mutations are in the at least one endogenous ACC synthase gene.
10. The plant cell according to claim 1, characterized in that the at least one interruption is introduced into the inactive plant cell by introducing at least one polynucleotide sequence comprising a nucleic acid sequence of ACC synthase, or subsequence of the same, in the inactive plant cell, such that at least one polynucleotide sequence is linked to a promoter in a sense or antisense orientation, and wherein the at least one polynucleotide sequence comprises at least about 85% of sequence identity to SEQ ID NO: 4 (cACS2), SEQ ID NO: 5 (cACSd), SEQ ID NO: 6 (cACS7) or SEQ ID NO: 10 (CCRA178R), or a subsequence thereof or a complement of it.
The plant cell according to claim 1, characterized in that the at least one interruption is introduced into the inactive plant cell by introducing at least one polynucleotide sequence comprising one or more subsequences of a nucleic acid sequence of ACC synthase configured for RNA muting or interference.
12. The plant cell according to claim 1, characterized in that the plant cell is of a dicot or monotone.
13. The plant cell according to claim 12, characterized in that the dicotyledone or monocotyledon is Zea mays, wheat, rice, sorghum, barley, oats, turf grass, rye, soybeans, Brassica, sunflower or cotton.
14. The plant cell according to claim 1, characterized in that the plant cell is in a plant that comprises a phenotype of green permanence potential.
15. The plant cell according to claim 1, characterized in that the plant cell is in a plant comprising a male sterility phenotype.
16. A plant, characterized in that it is regenerated from the plant cell of claim 1.
17. An inactive plant, characterized in that it comprises a phenotype of green permanence potential, the phenotype of green permanence potential that results from an interruption in at least one endogenous ACC synthase gene, wherein the interruption comprises one or more transposons or one or more point mutations, and wherein the disruption inhibits the expression or activity of at least one ACC synthase protein compared to a corresponding control plant.
18. The inactive plant according to claim 17, characterized in that the at least one endogenous ACC synthase gene comprises a nucleic acid sequence, or complement thereof, which comprises less about 85% sequence identity to SEQ ID NO: 1 (gACS2), SEQ ID NO: 2 (gACSβ) or SEQ ID NO: 3 (gACS7) or a complement thereof.
19. The inactive plant according to claim 17, characterized in that the phenotype of green permanency potential of the transgenic plant comprises increased drought resistance, increased time to maintain a photosynthetically active plant or senescence of the delayed leaf, compared to the corresponding control plant.
20. The inactive plant according to claim 17, characterized in that the inactive plant is a hybrid plant.
21. The inactive plant according to claim 17, characterized in that the inactive plant is a dicot or monotone.
22. The inactive plant according to claim 21, characterized in that the inactive plant is Zea mays, wheat, rice, sorghum, barley, oats, turf grass, rye, soybeans, Brassica, sunflower or cotton.
23. A transgenic inactive plant, characterized in that it comprises a phenotype of green permanency potential, the phenotype of green permanency potential that results from at least one introduced transgene that inhibits the synthesis of ethylene, wherein the at least one transgene introduced comprises a nucleic acid sequence encoding at least one ACC synthase, the nucleic acid sequence comprising at least about 85% sequence identity to SEQ ID NO: 1 (gACS2), SEQ ID NO: 2 ( gACSß), SEQ ID NO: 3 (gACS7), SEQ ID NO: 4 (cACS2), SEQ ID NO: 5 (cACSß), SEQ ID NO: 6 (cACS7) or SEQ ID NO: 10 (CCRA178R), or a subsequence thereof, a complement thereto, and is in a configuration that modifies a level of expression or activity of the at least one ACC synthase.
24. The transgenic inactive plant according to claim 23, characterized in that the configuration comprises an antisense, sense or silencing or RNA interference configuration.
25. The transgenic inactive plant according to claim 23, characterized in that the transgene comprises a preferred tissue promoter, a temporarily regulated promoter or an inducible promoter.
26. The transgenic inactive plant according to claim 23, characterized in that the phenotype of green permanence potential of the transgenic plant comprises increased drought resistance, increased time to maintain a photosynthetically active plant or senescence of the delayed leaf, compared to the corresponding control plant.
27. The transgenic inactive plant in accordance with claim 23, characterized in that the plant is a dicot or monotone.
28. The transgenic inactive plant according to claim 27, characterized in that the plant is Zea mays, wheat, rice, sorghum, barley, oats, turfgrass, rye, soybeans, Brassica, sunflower or cotton.
29. A transgenic plant, characterized in that it comprises a green permanency potential phenotype, the green permanence potential phenotype resulting from at least one introduced transgene that inhibits ethylene synthesis, wherein the at least one introduced transgene comprises a nucleic acid sequence encoding a subsequence of at least one ACC synthase, the at least one ACC synthase comprising at least about 85% sequence identity to SEQ ID NO: 7 (pACS2), SEQ ID NO : 8 (pACSβ), SEQ ID NO: 9 (pACS7) or SEQ ID NO: 11 (pCCRA178R), is in an RNA silencing or interference configuration, and modifies an expression level or activity of at least one ACC synthase .
30. The transgenic plant according to claim 29, characterized in that the phenotype of green permanence potential of the transgenic plant comprises increased drought resistance, increased time to maintain a photosynthetically active plant or leaf senescence retards, compared to a corresponding control plant.
31. The transgenic plant according to claim 29, characterized in that the plant is a dicot or monotone.
32. The transgenic plant according to claim 31, characterized in that the plant is Zea mays, wheat, rice, sorghum, barley, oats, turfgrass, rye, soybeans, Brassica, sunflower or cotton.
33. An isolated or recombinant polynucleotide, characterized in that it comprises a member selected from the group consisting of: (a) a polynucleotide, or a complement thereof, comprising at least about 85% sequence identity to SEQ ID NO: 1 (gACS2), SEQ ID NO: 2 (gACSβ), SEQ ID NO: 3 (gACS7), SEQ ID NO: 4 (cACS2), SEQ ID NO: 5 (cACSβ), SEQ ID NO: 6 (cACS7) or SEQ ID NO: 10 (CCR? 178R), or a subsequence thereof; (b) a polynucleotide, or a complement thereof, that encodes a polypeptide sequence of SEQ ID NO: 7 (pACS2), SEQ ID NO: 8 (pACSβ), SEQ ID NO: 9 (pACS7) or SEQ ID NO : 11 (pCCRA178R), or a subsequence thereof, or a conservative variation thereof; and (c) a polynucleotide, or a complement thereof, which hybridizes under severe conditions on substantially the entire length of a polynucleotide subsequence comprising at least 100 contiguous nucleotides of SEQ ID NO: 1 (gACS2), SEQ ID NO: 2 (gACSβ), SEQ ID NO: 3 (gACS7 ), SEQ ID NO: 4 (cACS2), SEQ ID NO: 5 (cACSβ), SEQ ID NO: 6 (cACS7) or SEQ ID NO: 10 (CCRA178R).
34. The isolated or recombinant polynucleotide according to claim 33, characterized in that the polynucleotide inhibits the production of ethylene when expressed in a plant.
35. An expression cassette, characterized in that it comprises a promoter operably linked to the isolated or recombinant polynucleotide of claim 33.
36. The expression cassette according to claim 35, characterized in that the promoter is a constitutive promoter.
37. The expression cassette according to claim 35, characterized in that the promoter is an inducible promoter or a temporarily regulated promoter.
38. The expression cassette according to claim 35, characterized in that the promoter is a preferred tissue promoter.
39. The expression limit in accordance with the claim 38, characterized in that the preferred tissue promoter is a preferred leaf promoter or a preferred anther promoter.
40. The expression cassette according to claim 35, characterized in that the polynucleotide is linked to the promoter in an antisense orientation.
41. The expression cassette according to claim 35, characterized in that the polynucleotide is linked to the promoter in a sense orientation.
42. The expression cassette according to claim 35, characterized in that the polynucleotide is in a configuration of RNA silencing or interference.
43. A vector, characterized in that it comprises a promoter operably linked to the isolated or recombinant polynucleotide of claim 33.
44. The vector according to claim 43, characterized in that the vector is a viral vector.
45. An inactive plant, characterized in that it comprises a male sterility phenotype, the male sterility phenotype resulting from at least one interruption in at least one endogenous ACC synthase gene, the interruption that inhibits the expression or activity of at least one an ACC synthase protein compared to a corresponding control plant, wherein the at least one ACC gene Endogenous synthase comprises a nucleic acid sequence, or complement thereof, comprising at least about 70% sequence identity to SEQ ID NO: 1 (gACS2), SEQ ID NO: 2 (gACSβ) or SEQ ID NO : 3 (gACS7).
46. The inactive plant according to claim 45, characterized in that the at least one interruption results in the production of reduced ethylene by the inactive plant cell as compared to the control plant.
47. The inactive plant according to claim 45, characterized in that the at least one interruption comprises one or more transposons, wherein the one or more transposons are in at least one endogenous ACC synthase gene.
48. The inactive plant according to claim 45, characterized in that the at least one interruption comprises one or more point mutations, wherein the one or more point mutations are in at least one endogenous ACC synthase gene.
49. The inactive plant according to claim 45, characterized in that the at least one interruption is introduced into the inactive plant by introducing at least one polynucleotide sequence comprising one or more sequences of an ACC synthase nucleic acid sequence. configured for muting or RNA interference.
50. The inactive plant according to claim 45, characterized in that the male sterility phenotype comprises the reduction of pollen reduced by the inactive plant as compared to the control plant.
51. An inactive transgenic plant, characterized in that it comprises a male sterility phenotype, the male sterility phenotype resulting from at least one introduced transgene that inhibits ethylene synthesis, wherein the at least one introduced transgene comprises a sequence of nucleic acid encoding at least one ACC synthase, the nucleic acid sequence comprising at least about 85% sequence identity to SEQ ID NO: 1 (gACS2), SEQ ID NO: 2 (gACSβ), SEQ ID NO: 3 (gACS7), SEQ ID NO: 4 (cACS2), SEQ ID NO: 5 (cACSβ), SEQ ID NO: 6 (CACS7) or SEQ ID NO: 10 (CCRA178R) or a subsequence thereof, or a complement thereto, and is in a configuration that modifies an expression level or activity of at least one ACC synthase.
52. The transgenic inactive plant according to claim 51, characterized in that the configuration comprises an antisense, sense or silencing or RNA interference configuration.
53. The inactive transgenic plant in accordance with claim 51, characterized in that the transgene comprises a preferred tissue promoter, a temporarily regulated promoter or an inducible promoter.
54. A method for inhibiting the production of ethylene in a plant, the method characterized in that it comprises: inactivating one or more ACC synthase genes in the plant, wherein the one or more ACC synthase genes encodes one or more ACC synthases, wherein at least one of the one or more ACC synthases comprises at least about 85% identity to SEQ ID NO: 7 (pACS2), SEQ ID NO: 8 (pACSβ), SEQ ID NO: 9 (pAC7) or SEQ ID NO: 11 (pCCRA178R).
55. The method according to claim 54, characterized in that the at least one of the one or more ACC synthase genes is at least 85% identical to SEQ ID NO: 1 (gACS2), SEQ ID NO: 2 ( gACS6) or SEQ ID NO: 3 (gAC7) or a complement thereof.
56. The method according to claim 54, characterized in that the step of inactivating comprises introducing one or more mutations into an ACC synthase gene sequence, wherein the one or more mutations in the ACC synthase gene sequence comprises one or more transposons , to thereby inactivate the one or more ACC synthase genes compared to a corresponding control plant.
57. The method according to claim 56, characterized in that the one or more mutations comprise a homozygous interruption in the one or more ACC synthase genes.
58. The method according to claim 56, characterized in that the one or more mutations comprise a heterozygous interruption in the one or more ACC synthase genes.
59. The method according to claim 56, characterized in that the one or more mutations are introduced by a sexual cross.
60. The method of compliance with the claim 54, characterized in that the inactivation step comprises introducing one or more mutations into an ACC synthase gene sequence, wherein the one or more mutations in the ACC synthase gene sequence comprises one or more point mutations, in order to inactivate the one or more ACC synthase genes compared to a corresponding control plant.
61. The method of compliance with the claim 54, characterized in that the inactivation step comprises: (a) introducing into the plant at least one polynucleotide sequence, wherein the at least one polynucleotide sequence comprises a nucleic acid encoding one or more ACC synthases, or a subsequence of it, and a promoter, the promoter that works in plants to produce an RNA sequence; and (b) expressing at least one polynucleotide sequence, thereby inactivating the one or more ACC synthase genes compared to a corresponding control plant.
62. The method according to claim 61, characterized in that the at least one polynucleotide sequence is introduced by electroporation, micro-projectile bombardment or Agrobacterium-mediated transfer.
63. The method according to claim 61, characterized in that the polynucleotide is linked to the promoter in a sense orientation.
64. The method of compliance with the claim 61, characterized in that the polynucleotide is linked to the promoter in an antisense orientation.
65. The method according to claim 61, characterized in that the polynucleotide is configured for RNA silencing or interference. ßß.
The method in accordance with the claim 61, characterized in that the promoter is a preferred tissue promoter, a temporarily regulated promoter or an inducible promoter.
67. The method according to the claim 54, characterized in that the plant is a dicot or monotone.
68. The plant according to claim 67, characterized in that the plant is Zea mays, wheat, rice, sorghum, barley, oats, turf grass, rye, soybeans, Brassica, sunflower or cotton.
69. A plant, characterized in that it is produced by the method of claim 54.
70. A method modulates the green residence potential in a plant, the method characterized in that it comprises: a) selecting at least one ACC synthase gene to mutate, the at least one ACC synthase gene comprising a nucleic acid sequence, or complement thereof, comprising at least about 70% sequence identity to SEQ ID NO: 1 (gACS2), SEQ ID NO: 2 (gACSd) or SEQ ID NO: 3 (gACS7), to thereby provide at least one desired ACC synthase gene; b) introducing a mutant form of at least one desired ACC synthase gene into the plant; and c) express the mutant form, in order to modulate the potential for green permanence in the plant.
71. The method according to claim 70, characterized in that the green remaining potential of the plant comprises: (a) a reduction in the production of at least one specific mRNA of ACC synthase; (b) a reduction in the production of an ACC synthase; (c) a reduction in ethylene production; (d) a delay in the senescence of the leaf; (e) an increase in drought resistance; (f) an increased time in maintaining the photosynthetic activity; (g) increased perspiration; (h) an increased stomatal conductance; (i) an assimilation of C02 increased; (j) an increased time in maintaining the assimilation of CO2; or, (k) any combination of (a) - (j); compared to a corresponding control plant.
72. The method of compliance with the claim 70, characterized in that the selection of at least one gene ACC synthase comprises determining a desired green residence potential degree.
73. The method according to claim 72, characterized in that the degree of permanency potential desired green is weak, moderate or strong.
74. The method according to claim 70, characterized in that the ACC synthase gene encodes an ACC synthase selected from the group consisting of: SEQ ID NO: 7 (pACS2), SEQ ID NO: 8 (pACSβ), SEQ ID NO: 9 (pAC7) and SEQ ID NO: 11 (pCCRA178R).
75. The method according to claim 70, characterized in that the mutant form comprises a heterozygous mutation in at least one ACC synthase gene.
76. The method of compliance with the claim 70, characterized in that the mutant form comprises a mutation homozygous in at least one ACC synthase gene.
77. The method according to claim 70, characterized in that the mutant form comprises subsequence of at least one desired ACC synthase gene in an antisense, sense or silencing or RNA interference configuration.
78. The method according to the claim 70, characterized in that the mutant form is introduced by Agrobacterium-mediated transfer, electroporation, micro-projectile bombardment or a sexual cross.
79. The method according to claim 70, characterized in that the plant is a dicot or monotone.
80. The plant according to claim 79, characterized in that the plant is Zea mays, wheat, rice, sorghum, barley, oats, turf grass, rye, soybeans, Brassica, sunflower or cotton.
81. A plant, characterized in that it is produced by the method of claim 70.
82. A device for modulating the green residence potential in a plant, the device characterized in that it comprises: at least one polynucleotide sequence comprising a sequence of nucleic acid, wherein the nucleic acid sequence is at least about 85% identical to SEQ ID NO: 1 (gACS2), SEQ ID NO: 2 (gACSd), SEQ ID NO: 3 (gACS7), SEQ ID NO : 4 (cACS2), SEQ ID NO: 5 (cACS6), SEQ ID NO: 6 (cAC7) or SEQ ID NO: 10 (CCRA178R) or a subsequence thereof, or a complement thereto.
83. The equipment according to claim 82, characterized in that the equipment further comprises instructional materials for the use of at least one polynucleotide sequence to control the green residence potential in a plant.
84. A device for modulating male sterility in a plant, the device characterized in that it comprises: at least one polynucleotide sequence comprising a nucleic acid sequence, wherein the acid sequence nucleic acid is at least approximately 85% identical to SEQ ID NO: 1 (gACS2), SEQ ID NO: 2 (gACSβ), SEQ ID NO: 3 (gACS7), SEQ ID NO: 4 (cACS2), SEQ ID NO: 5 (cACSß), SEQ ID NO: 6 (cAC7) or SEQ ID NO: 10 (CCRA178R), or a subsequence thereof, or a complement thereto.
85. The equipment according to claim 84, characterized in that the equipment further comprises instructional materials for the use of at least one polynucleotide sequence to control male sterility in a plant.