CN101040050A - Plants having improved growth characteristics and method for making the same - Google Patents

Abstract

The present invention concerns a method for improving the growth characteristics of plants by increasing activity in a plant of an RNA-binding protein or a homologue thereof, wherein said RNA-binding protein or homologue thereof is either: (i) a polypeptide having RNA-binding activity and comprising either 2 or 3 RNA 10 recognition motifs (RRMs) and a motif having at least 75% sequence identity to motif I: PIYEAAVVALPVVVKERLVRILRLGIATRYD and/or a motif having at least 50% sequence identity to motif II: RFDPFTGEPYKFDP; or (ii) an RBP1 polypeptide or homologue thereof having (a) RNA-binding activity; (b) two RRM domains, (c) the following two motifs: (i) KIFVGGL; and (ii) 15 RPRGFGF, allowing for up to three amino acid substitutions and any conservative change in the motifs; and (d) having at least 20% sequence identity to the amino acid represented by SEQ ID NO: 15. The invention also concerns to transgenic plants having introduced therein an RNA-binding protein-encoding nucleic acid or variant thereof, which plants have improved growth characteristics relative to corresponding wild type plants. The present invention also concerns constructs useful in the methods of the invention.

Description

Has plant of improvement growth characteristics and preparation method thereof

Technical field

Relate generally to biology field of the present invention, and relate to the method that improves plant growth characteristics.More specifically, the present invention relates to improve plant growth characteristics, especially the method for productive rate by the activity of rna binding protein or its homologue in the increase plant.The invention still further relates to the plant with the active rna binding protein of increase or its homologue, these plants have the growth characteristics of improvement with respect to corresponding wild type plant.Rna binding protein or its homologue useful in the inventive method have RNA in conjunction with activity, and comprise 2 or 3 RNA identification motifs (RRM), and comprise with motif I:PYEAAVVALPVVVKERLVRILRLGIATRYD and have the motif of at least 75% sequence identity and/or the motif that has at least 50% sequence identity with motif II:RFDPFTGEPYKFDP.Rna binding protein or its homologue useful in the inventive method also can be RBP1 polypeptide or its homologues with following feature: (a) RNA is in conjunction with activity; (b) two RRM structural domains; (c) following two motifs: (i) KIFVGGL and (ii) RPRGFGF allow three aminoacid replacement and conservative arbitrarily change the at the most in the motif; (d) amino acid according to preference ordering that increases and SEQ IDNO:15 representative has at least 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95% sequence identity.The present invention also provides useful in the methods of the invention construct.

Technical background

The continuous growth of world population and be used for the minimizing of tilled the land supplys of agricultural promotes farming research and develops towards the direction of raising farm efficiency.The ordinary method of farm crop and gardening improvement is the plant that utilizes breeding technique to identify to have desired characteristic.Yet this breeding technique has several shortcomings, and promptly these technology are normally labor-intensive, and produces the plant that contains the allogeneic heredity component usually, and these hereditary components are not always to produce the desirable proterties of transmitting from mother plant.Molecular biological progress has allowed the germplasm (germplasm) of human reconstruction animal and plant.Plant genetic engineering need separate and operate genetic material (common form with DNA or RNA) and reach subsequently with this genetic material introduced plant.This techniques enable is enough sent the farm crop or the plant of economy, agricultural or gardening proterties with multiple improvement.A proterties with special economic interests is a productive rate.Productive rate is generally defined as from the economic worth of farm crop can measure output.It can define according to quantity and/or quality.Productive rate directly depends on several factors, for example: the quantity of organ and size, plant structure (for example branch amount), seed production and more factors.The growth of root, dietetic alimentation and stress tolerance also are the important factors of decision productive rate.Can increase the productive rate of farm crop by optimizing one of above-mentioned factor.

The ability of the multiple growth characteristics of improvement plant will as: the fields such as production, arboriculture (aboriculture), Horticulture and forestry that increase farm crop, plant breeding, ornamental plant have many application.The growth characteristics of improvement, as productive rate, the algae that also can be used for using at bio-reactor produces (be used for the biotechnology production such as medicine, antibody or vaccine substance, or be used for the bio-transformation of organic waste) and other this class field.

Have now found that, compare with corresponding wild-type plant, the activity of rna binding protein or its homologue makes plant have the growth characteristics of improvement in the increase plant, described rna binding protein or its homologue have RNA in conjunction with active and comprise 2 or 3 RNA identification motifs (RRM), and comprise with motif I:PYEAAVVALPVVVKERLVRILRLGIATRYD and have the motif of at least 75% sequence identity and/or the motif that has at least 50% sequence identity with motif II:RFDPFTGEPYKFDP.Have now found that, compare that the activity of RBP1 polypeptide or its homologue makes plant have the growth characteristics of improvement in the increase plant with corresponding wild-type plant.The polypeptide that described RBP1 polypeptide or its homologue refer to have following feature: (a) RNA is in conjunction with activity; (b) two RRM structural domains; (c) following two motifs: (i) KIFVGGL and (ii) RPRGFGF allow three aminoacid replacement and conservative arbitrarily change the at the most in the motif; (d) amino acid according to preference ordering that increases and SEQ ID NO:15 representative has at least 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95% sequence identity.

Rna binding protein has vital role in the adjusting of transcribing with post-transcriptional level genetic expression.Synthetic, processing of regulating that extends horizontally to the RNA molecule and the institute in the turnover comprise premessenger RNA montage, polyadenylation, mRNA transhipment, translation and stability/decay in steps.Regulating mainly is directly to finish by rna binding protein, or the function of regulating other regulatory factor by rna binding protein is indirectly finished.The RNA-protein interaction is many-sided key link that cellular metabolism, cytodifferentiation and growth and pathogen infection duplicate.RNA identification motif or RRM are present in the multiple rna binding protein usually and relate to all and transcribe post-treatment, and wherein from one to four copy of each proteinic RRM number changes.RRM is about 80 amino acid whose zones of containing several high conservative residues, and it is the subunit preface of two weak points that wherein some are gathered, RNP-1 (eight aggressiveness) and RNP-2 (six aggressiveness) (Birney etc., Nucleic Acids Research, 1993, Vol.21, No.25,5803-5816).

196 protein that contain RRM of Arabidopis thaliana (Arabidopsis) genome encoding, the example be RBP1 (Lorkovic etc., Nucleic Acids Research, 2002, Vol.30, No.3,623-635).They report that the RRM of AtRBP1 is the most similar to the proteinic RRM of metazoan Musashi.Except AtRBP1, Lorkovic etc. have also described three protein that have similarity with AtRBP1 and Musashi protein.RBP1 from Arabidopis thaliana (Arabidopsis thaliana) separates expression in the concurrent now quick splitted tissue first by (Plant Cell Physiol.41 (3): 282-288 (2000)) such as Suzuki.RBP1 is a kind of rna binding protein (shown in Suzuki etc. 2000), comprises two RRM.

Summary of the invention

According to one embodiment of the invention, the method of improvement plant growth characteristics is provided, it comprises the activity that increases rna binding protein in the plant or its homologue, described rna binding protein or its homologue have RNA and discern motifs (RRM) in conjunction with active and 2 or 3 RNA, and comprise with motif I:PYEAAVVALPVVVKERLVRILRLGIATRYD and have the motif of at least 75% sequence identity and/or the motif that has at least 50% sequence identity with motif II:RFDPFTGEPYKFDP.

According to one embodiment of the invention, the method for improvement plant growth characteristics is provided, it comprises the activity that increases RBP1 polypeptide in the plant or its homologue, and described RBP1 polypeptide or its homologue have following feature: (a) RNA is in conjunction with activity; (b) two RRM structural domains; (c) following two motifs: (i) KIFVGGL; (ii) RPRGFGF allows three aminoacid replacement and conservative arbitrarily change at the most in the motif; (d) amino acid according to preference ordering that increases and SEQ ID NO:15 representative has at least 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95% sequence identity.

Advantageously, carry out method of the present invention and make plant have the growth characteristics of multiple improvement, especially the productive rate of Zeng Jiaing, particularly seed productive rate.

Term defined herein " productive rate of increase " means with respect to corresponding wild type plant, increase in following arbitrary or many aspects: (i) one or more parts of plant, particularly the biomass (weight), the root biomass that increases or any other that increase of (can gather in the crops) part can be gathered in the crops the biomass that part increases on the ground; (ii) the seed productive rate of Zeng Jiaing comprises the increase of seed biomass (seed weight), and it can be in every strain plant seed weight or based on the increase of single seed; (iii) (full) seed amount of Zeng Jiaing; The (iv) seed size of Zeng Jiaing, it also may influence the composition of seed; (the v) seed volume of Zeng Jiaing, it also may influence the composition of seed; (the vi) harvest index of Zeng Jiaing, it is expressed as the productive rate that can gather in the crops part (as seed) and the ratio of total biomass; (the vii) thousand seed weight of Zeng Jiaing (TKW), it is to calculate from the gross weight of full seed number.The TKW that increases may be derived from the seed size and/or the seed weight of increase.

With the corn is example, and the increase of productive rate can show as following one or more: the increase of per hectare or every acre of plant quantity, the increase of every strain plant spike number, increase of line number, a row grain number, grain weight, thousand seed weight, fringe length/diameter or the like.With the rice is example, and the increase of productive rate can show as following one or more: the quantity of per hectare or every acre of plant, the panicle number of every strain plant, each paniculiform spikelet number, each paniculiform number of spending, the increase of the full rate of seed, increase of thousand seed weight or the like.The increase of productive rate is the mutagenic structure of possibility also, and the result who perhaps can be used as the structure of change takes place.

According to preferred aspect, carry out method of the present invention and produce plant with increase productive rate.Therefore, the invention provides the method that increases plant yield, this method comprises the activity that increases rna binding protein in the plant or its homologue, described rna binding protein or its homologue have RNA and discern motifs (RRM) in conjunction with active and 2 or 3 RNA, and comprise with motif I:PYEAAVVALPVVVKERLVRILRLGIATRYD and have the motif of at least 75% sequence identity and/or the motif that has at least 50% sequence identity with motif II:RFDPFTGEPYKFDP.According to another preferred aspect of the present invention, the method that increases plant yield is provided, described method comprises the activity that increases RBP1 polypeptide in the plant or its homologue, described RBP1 polypeptide or its homologue have following feature: (a) RNA is in conjunction with activity; (b) two RRM structural domains; (c) following two motifs: (i) KIFVGGL and (ii) RPRGFGF allow three aminoacid replacement and conservative arbitrarily change the at the most in the motif; (d) amino acid according to preference ordering that increases and SEQ ID NO:15 representative has at least 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95% sequence identity.

Because transgenic plant of the present invention have the productive rate of increase, for the growth velocity of their life cycle respective stage, these plants may present the growth velocity (in life cycle of their parts at least) of increase with respect to corresponding wild-type plant.The growth velocity that increases may be specific to one or more parts (comprising seed) of plant, perhaps may spread all over whole plants basically.Have the plant that increases growth velocity even may present early flowering.The increase of growth velocity may appear at one or more stages in plant life cycle, perhaps appears in the process of whole plants life cycle basically.At the commitment in plant life cycle, the growth of growth velocity may show as the enhanced vigor.The increase of growth velocity can change the harvest cycle of plant, makes the plant can be than other possible situation more late sowing kind and/or faster results.If growth velocity fully increases, may allow to sow the more seed of kindred plant species (for example sow and gather in the crops rice plants, subsequently in sowing in vegetative period of a routine and gather in the crops more rice plants).Same, if growth velocity increases fully, may allow to sow the more seed of different plant species (for example sow and gather in the crops rice plants, subsequently, as sowing and optional results soybean, potato or any plant that other is fit to).Also may be under the situation of some plants from the extra number of times of same rhizome results.The harvest cycle that changes plant may cause every acre year biomass yield increase (because increase of (for example in 1 year) any specified plant growth and harvesting frequency).Compare with wild type counterparts, the increase of growth velocity also may allow more wide region cultivation transgenic plant because the region restriction of plantation farm crop during usually by plantation when (season early) or results (season in evening) hostile environment condition determined.If the shortening harvest cycle can be avoided this class unfavourable condition.Can determine growth velocity from a plurality of parameters the growth curve that growth experiment is drawn, this class parameter can be: T-Mid (plant reaches 50% required time of its largest amount) and T-90 (plant reaches its largest amount 90% required time) or the like.

Carrying out method of the present invention makes plant have the growth velocity of increase.Therefore, the invention provides the method that increases plant growth rate, this method comprises the activity that increases rna binding protein in the plant or its homologue, described rna binding protein or its homologue have RNA and discern motifs (RRM) in conjunction with active and 2 or 3 RNA, and comprise with motif I:PYEAAVVALPVVVKERLVRILRLGIATRYD and have the motif of at least 75% sequence identity and/or the motif that has at least 50% sequence identity with motif II:RFDPFTGEPYKFDP.Another method that increases plant growth rate also is provided, and described method comprises the activity that increases RBP1 polypeptide in the plant or its homologue, and described RBP1 polypeptide or its homologue have following feature: (a) RNA is in conjunction with activity; (b) two RRM structural domains; (c) following two motifs: (i) KIFVGGL and (ii) RPRGFGF allow three aminoacid replacement and conservative arbitrarily change at the most in the motif; (d) amino acid according to preference ordering that increases and SEQ ID NO:15 representative has at least 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95% sequence identity.

Be under the no stress conditions plant, or be exposed to respect to control plant and multiplely slightly coerce down, the increase of productive rate and/or growth velocity takes place in it.Usually plant is replied to be exposed to by growth more slowly and coerces.Under the severe stress conditions, plant even can stop growing fully.On the other hand, slightly coerce to be defined as in this article when plant and be exposed to this, do not cause the plant forfeiture that stops growing fully to restart any of energy for growth and coerce.Because the development of the farming method (irrigation, fertilising, pesticide treatments), the crop plants of cultivation usually can not run into severe and coerce.Therefore, by the impaired feature of not expecting in the agricultural that is grown to serve as of slight stress-inducing.Slightly coerce is that the typical case that plant may contact coerces.These coerce may be the plant daily biology that is exposed to it and/or abiotic (environment) coerce.The coercing of typical abiotic or environment comprise by the temperature that the heat of abnormality or cold/freezing temperature produces coerce, salt stress, water coerce (arid or excessive water).Abiotic stress also can be caused by pharmaceutical chemicals.Biological coercing generally is that those that caused by pathogenic agent such as bacterium, virus, fungi and insect are coerced.

Can in any plant, advantageously modify above-mentioned growth characteristics.

Ancestors of whole plants, plant and the part of offspring and plant contained in term used herein " plant ", comprises seed, bud, stem, leaf, root, flower (comprising stem tuber) and tissue and organ, and wherein above-mentioned every kind comprises goal gene/nucleic acid.Suspension culture, callus, embryo, meristematic zone, gametophyte, sporophyte, pollen and sporule also contained in term " plant ", and wherein above-mentioned every kind equally also comprises goal gene/nucleic acid.

The plant that can be used for especially in the inventive method comprises the whole plants that belong to vegitabilia (Viridiplantae) superfamily, particularly unifacial leaf and dicotyledons comprise the feed or the feed leguminous plants that are selected from following species, ornamental plant, food crop, arbor or shrub: Acacia species (Acaciaspp.), maple species (Acer spp.), Actinidia species (Actinidia spp.), Aesculus species (Aesculus spp.), New Zealand kauri (Agathis australis), Albiziaamara, Alsophila tricolor, Andropogon species (Andropogon spp.), Arachis species (Arachis spp.), betel nut (Areca catechu), Asteliafragrans, Astragaluscicer, Baikiaea plurijuga, Betula (Betula spp.), Btassica (Brassica spp.), Bruguiera conjugata (Bruguiera gymnorrhiza), Burkea africana, palas (Butea frondosa), Cadaba farinosa, Zhu Ying Pittosporum species (Calliandra spp.), daye tea (Camelliasinensis), Canna generalis Bailey (Canna indica), Capsicum species (Capsicum spp.), Cassia species (Cassia spp.), apart from pea (Centroema pubescens), Chaenomeles species (Chaenomeles spp.), Chinese cassia tree (Cinnamomum cassia), fruitlet coffee (Coffeaarabica), Colophospermum mopane, variation coronule flower (Coronillia varia), Cotoneaster serotina, hawthorn species (Crataegus spp.), Cucumis species (Cucumisspp.), Cupressus species (Cupressus spp.), Cyathea dealbata Quinces Quince (Cydoniaoblonga), Japanese cypress (Cryptomeria japonica), Cymbopogon species (Cymbopogonspp.), Cynthea dealbata Quinces Quince (Cydonia oblonga), Dalbergia monetaria, Da Ye Rhizome of Fortune's Drynaria (Davallia divaricata), mountain horseleech species (Desmodium spp.), coarse freshwater mussel shellfish fern (Dicksonia squarosa), Diheteropogon amplectens, Dioclea spp, sickle Dolichos species (Dolichos spp.), Dorycnium rectum, Echinochloapyramidalis, Ehrartia spp. Finger-millet (Eleusine coracana), Herba Eragrostidis pilosae species (Eragrestis spp.), Erythrina species (Erythrina spp.), eucalyptus species (Eucalyptusspp.), Euclea schimperi, Eulalia villosa, Fagopyrum species (Fagopyrum spp.), feijoa (Feijoa sellowiana), Fragaria species (Fragaria spp.), Moghania species (Flemingia spp), Freycinetia banksii, Geranium thunbergii, ginkgo (Ginkgobiloba), Glycine javanica, Gliricidia spp, upland cotton (Gossypium hirsutum), Grevillea species (Grevillea spp.), Guibourtia coleosperma, rock Astragalus species (Hedysarum spp.), Hemarthria compressa (Hemarthia altissima), turn round Huang Mao (Heteropogoncontortus), barley (Hordeum vulgare), Hyparrhenia rufa, Herba Hyperici Erecti (Hypericumerectum), Hyperthelia dissoluta, spend front yard indigo plant (Indigo incarnata) in vain, Jris species (Iris spp.), Leptarrhena pyrolifolia, lespedeza species (Lespediza spp.), Lettuca spp., Leucaena leucocephala, Loudetia simplex, sieve beans (Lotonusbainesii) that pause, Lotus species (Lotus spp.), Macrotyloma axillare, Malus species (Malus spp.), Manihot esculenta, alfalfa (Medicago sativa), metasequoia (Metasequoia glyptostroboides), powder bajiao banana (Musa sapientum), Nicotiana species (Nicotianum spp.), donkey food Macroptilium species (Onobrychis spp.), Ornithopus spp., Oryza species (Oryza spp.), Peltophorum africanum, Pennisetum species (Pennisetumspp.), Persea gratissima, green winter Solanum species (Petunia spp.), Phaseolus species (Phaseolus spp.), betel nut bamboo (Phoenix canariensis), Phormium cookianum, Photinia species (Photinia spp.), white spruce (Piceaglauca), Pinus species (Pinusspp.), pea (Pisum sativum), alpine totara (Podocarpus totara), Pogonarthria fleckii, Pogonarthria squarrosa, Populus species (Populus spp.), algarroba (Prosopis cineraria), Pseudotsuga menziesii (Mirbel) Franco (Pseudotsuga menziesii), Pterolobiumstellatum, European pear (Pyrus communis), oak species (Quercus spp.), Rhaphiolepsis umbellata, delicious rod is spent palm fibre (Rhopalostylis sapida), Rhusnatalensis, Europe gooseberry (Ribes grossularia), currant species (Ribes spp.), acacia (Robinia pseudoacacia), rose species (Rosa spp.), rubus species (Rubus spp.), Salix species (Salix spp.), Schyzachyrium sanguineum, parasol pine (Sciadopitys verticillata), sequoia sempervirens (Sequoia sempervirens), big tree (Sequoiadendron giganteum), dichromatism chinese sorghum (Sorghum bicolor), spinach species (Spinacia spp.), Sporobolus fimbriatus, Stiburus alopecuroides, Stylosanthos humilis, Triquetrous Tadehagi Herb species (Tadehagi spp.), southern cypress (Taxodiumdistichum), Arabic Herba Themedae japonicae (Themeda triandra), Clover species (Trifoliumspp.), Triticum species (Triticum spp.), tsuga heterophylla (Tsuga heterophylla), genus vaccinium species (Vaccinium spp.), Vetch species (Vicia spp.), grape (Vitisvinifera), the fertile gloomy flower (Watsonia pyramidata) of awl fringe, common calla (Zantedeschiaaethiopica), corn (Zea mays), Amaranthus, arithoke, asparagus, cabbage, brassica oleracea var gemmifera, Caulis et Folium Brassicae capitatae, rape, Radix Dauci Sativae, Cauliflower, celery, the green wild cabbage of the garment or robe made of feathers, flax, kale; root of Szemao crotalaria; oil grain rape; gumbo; onion; potato; rice; soybean; strawberry; sugar beet; sugarcane; Sunflower Receptacle; tomato; pumpkin; tea tree and algae or the like.According to the preferred embodiment of the invention, described plant is a crop plants, as soybean, Sunflower Receptacle, rape, clover, Semen Brassicae campestris, cotton, tomato, potato or tobacco.Further preferred plant is a monocotyledons, as sugarcane.Preferred plant is a cereal, as rice, corn, wheat, barley, grain, rye, Chinese sorghum or oat.

Can increase the activity of rna binding protein or its homologue by the level that improves rna binding protein in the plant.Alternatively, when the rna binding protein level does not increase, perhaps even when the rna binding protein level reduces, also can increase its activity.When this situation appears at the polypeptide natural characteristics and changes, for example, have more active mutant forms than wild type peptide by preparation.Similarly, can increase the activity of RBP1 polypeptide or its homologue by the level that improves RBP1 polypeptide protein in the plant.Alternatively, when the RBP1 level does not change, perhaps even when RBP1 polypeptide level reduces, also can increase its activity.When this situation appears at the polypeptide natural characteristics and changes, for example, have more active mutant than wild-type by preparation.

The term " rna binding protein or its homologue " of this paper definition refers to have RNA and discerns the polypeptide of motifs (RRM) in conjunction with active and 2 or 3 RNA, and comprises with motif I:PYEAAVVALPVVVKERLVRILRLGIATRYD and have the motif of at least 75%, 80%, 85%, 90% or 95% sequence identity and/or the motif that has at least 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90% or 95% sequence identity with motif II:RFDPFTGEPYKFDP.This term also refers to such aminoacid sequence, and described aminoacid sequence has at least 13%, 15%, 17%, 19%, 21%, 23%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90% or 95% sequence identity according to the aminoacid sequence of preference ordering that increases and SEQ ID NO:2 representative.

Can use routine techniques well-known in the art easily to identify " rna binding protein or its homologue " in the above range of definition.For example, use technology well-known in the art can determine easily in external or body that RNA is in conjunction with activity.The example of external test comprises: use nucleic acid that North-Western and/or South-Western analyze in conjunction with measuring .PlantCell Physiol.41 (3): 282-288 (2000) such as () Suzuki; Use the crosslinked RNA of UV in conjunction with mensuration; The electrophoretic mobility shift assay of rna binding protein (Smith, RNA-Protein Interactions-APractical Approach 1998, University of Cambridge).The example of measuring in the body comprises TRAP (translation repression measuring method) (Paraskeva E, Atzberger A, Hentze MW:Atranslational repression assay procedure (TRAP) for RNA-proteininteractions in vivo.PNAS 1998Feb 3; 95 (3): 951-6.).

Can determine easily whether the amino acid of polypeptide and SEQ ID NO:2 representative has at least 13% identity by sequence alignment.The method of sequence alignment is well-known in the art, and these class methods comprise GAP, BESTFIT, BLAST, FASTA and TFASTA.The comparison that the algorithm (J.Mol.Biol.48:443-453,1970) of GAP application Needleman and Wunsch is searched two complete sequences minimizes maximization of coupling number and room number.The per-cent of BLAST arithmetic calculation sequence identity, and the similarity between two sequences carried out statistical study.The software of carrying out the BLAST analysis can obtain publicly from biotechnology infonation center.For example use the VNTIAlignX multiple ratio to program, by the comparison (see for example shown in Figure 1 comparison) of search sequence (preferred protein sequence) with known rna binding protein sequence, can differentiate easily that the aminoacid sequence with SEQ ID NO:2 representative has rna binding protein or its homologue of at least 13% identity, described VNTI AlignX multiple ratio is to the clustal W algorithm (InforMax of program based on modification, Bethesda, MD, http://www.informaxinc.com), have the open point penalty in room be 10 and the room extend to 0.05 default setting.

The person skilled in the art can also easily identify with motif I:PYEAAVVALPVVVKERLVRILRLGIATRYD to have the motif of at least 75%, 80%, 85%, 90% or 95% sequence identity and/or the motif that has at least 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85% or 90% sequence identity with motif II:RFDPFTGEPYKFDP.This can be by comparing and searching for the homology zone and easily realize.

Following table 1 be presented at the motif I that finds among the sequence SEQ ID NO:2 and II and with cognate rna conjugated protein in the sequence identity percentage ratio of corresponding motif.Useful in the method for the invention rna binding protein may contain motif I or II, or motif I and II.

Table 1: the motif that in rna binding protein and its homologue, finds.

The example that falls into the polypeptide of " rna binding protein or its homologue " definition comprises following sequence: SEQ ID NO:2 is from tobacco; SEQ ID NO:4 is the protein prediction from rice BAC clone (NCBI accession number AL731884); SEQ ID NO:6 is the rice protein prediction (fragment) from cDNA (NCBI accession number AK059444); SEQ ID NO:8 is the corn protein prediction (fragment) from cDNA (NCBI accession number AY105295); And SEQ ID NO:10 is the rice sequence (NCBI accession number BAC83046) of total length.

Term " RBP1 or its homologue " polypeptide that refers to have following feature as definition here: (a) RNA is in conjunction with activity; (b) two RRM structural domains; (c) following two motifs: (i) KIFVGGL and (ii) RPRGFGF allow three aminoacid replacement and conservative arbitrarily change at the most in the motif; (d) amino acid according to preference ordering that increases and SEQ ID NO:15 representative has at least 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95% sequence identity.Conservative replacement table is (for example sees Creighton (1984) Proteins.W.H.Freeman and Company and see the following form 4) well-known in the art.

Can use the well-known routine techniques of those skilled in the art easily to identify to belong to " RBP1 polypeptide or its homologue " of above definition.For example, can determine easily as mentioned above that RNA is in conjunction with activity.

In addition, the RRM structural domain is well-known in the art and is made up of about 80-90 amino acid; They have and comprise four chains arranging with α/β interlayer form and the structure of two spirals, sometimes RNA in conjunction with in have the 3rd spiral.The protein that contains the RRM structural domain has mode configuration.Can use SMART to identify RRM structural domain (a Simple Modular ArchitectureResearch Tool:Identification of signaling domains, .PNAS such as Schultz, 95,5857-5864 (1998)) (http://smart.embl-heidelberg.de/).See Letunic etc. again, and Recent improvements to the SMART domain-based sequence annotationresource (Nucleic Acids Res.30 (1), 242-244).

The method that can use as mentioned above comparison determines easily that by sequence alignment whether polypeptide has at least 20% identity with the amino acid of SEQ ID NO:2 representative.

Because the RBP1 polypeptide comprises the zone of high conservative, those skilled in the art will be by can easily identifying those comparisons of the conserved regions of any search sequence and known RBP1 sequence other RBP1 sequence.The example of these conserved regions comprises following two motifs: (i) KIFVGGL and (ii) RPRGFGF allow three aminoacid replacement and conservative arbitrarily change at the most in the motif.

The example of polypeptide that belongs to " RBP1 polypeptide or its homologue " definition comprises: all from At1g58470 (SEQ ID NO:15), At4g26650 (SEQ ID NO:17), At5g55550 (SEQ ID NO:19), At4g14300 (SEQ ID NO:21), At3g07810 (SEQ ID NO:23), At2g33410 (SEQ ID NO:25) and the At5g47620 (SEQ ID NO:27) of Arabidopis thaliana; NP_921939.1 (SEQ ID NO:29) from rice; AK067725 (SEQ ID NO:31) and AK070544 (SEQ ID NO:33) corresponding to the RBP1 polypeptide of rice mRNA coding; From the CK210974 (SEQ ID NO:35) of wheat with from the CA124210 (SEQ IDNO:37) of sugarcane is partial protein prediction from EST (expressed sequence tag).

Although show low relatively sequence homology (approximately being low to moderate 25%), the full length protein that RPB1 protein and all have 2 RRM structural domains structurally is a high conservative.Can easily find the rbp1 gene (see above example from rice, sugarcane and wheat, it is accredited as RBP1 protein here for the first time) in other plant species thus.Following table 2 is presented at the identity percentage ratio of some sequences that show in the comparison of Fig. 3.

Table 2:RBP1 protein sequence and SEQ ID NO:2 are based on the homology of comprehensive whole sequence alignment

MIP accession number identifier ( http://mips.gsf.de/)	SEQ ID NO	The RRM structural domain	Whole homology VNTI comparison program (informax)
				At4g26650	SEQ ID NO：17	2X RRM	28.4％
At5g55550	SEQ ID NO：19	2X RRM	28.9％

At4g14300	SEQ ID NO:21	2X RRM	31.9%
				At3g07810	SEQ ID NO:23	2X RRM	24.9%
At2g33410	SEQ ID NO:25	2X RRM	29.2%
				At5g47620	SEQ ID NO:27	2X RRM 2X RRM	26.7%
AK070544-Os (corresponding to the dna sequence dna of mRNA).Chromosome position: BAC AC125782.2 (138541-142744)	SEQ ID NO:33	2X RRM	26.8%
				AK067725-OS (corresponding to the dna sequence dna of mRNA).Chromosome position: BAC AP003747 (103016-107790)	SEQ ID NO:31	2X RRM	26.3%

It should be understood that, term RBP1 polypeptide or its homologue are not limited to the sequence by SEQ ID NO:15, SEQ ID NO:17, SEQ ID NO:19, SEQ ID NO:21, SEQ ID NO:23, SEQ ID NO:25 or SEQ ID NO:27, SEQ ID NO:29, SEQ ID NO:31, SEQID NO:33, SEQ ID NO:35 and SEQ ID NO:37 representative, but meeting any polypeptide of following standard, described standard is: (a) RNA is in conjunction with activity; (b) two RRM structural domains; (c) following two motifs: (i) KIFVGGL and (ii) RPRGFGF allow three aminoacid replacement and conservative arbitrarily change at the most in the motif; (d) amino acid according to preference ordering that increases and SEQ IDNO:15 representative has at least 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95% sequence identity, and described polypeptide is useful in carrying out method of the present invention.

Coding RNA nucleic acid conjugated protein or its homologue can be any natural or synthetic nucleic acid.Rna binding protein or its homologue as above this paper definition are by rna binding protein coding nucleic acid/genes encoding.Therefore the term " rna binding protein coding nucleic acid/gene " as definition here is coding as the rna binding protein of above this paper definition or any nucleic acid/gene of its homologue.The example of rna binding protein coding nucleic acid comprises by those of arbitrary representative among SEQ ID NO:1, SEQ ID NO:3, SEQID NO:5, SEQ ID NO:7 and the SEQ ID NO:9.Rna binding protein coding nucleic acid/gene and functional variant thereof are applicable to implementing method of the present invention.The functional variant of rna binding protein coding nucleic acid/gene comprise rna binding protein coding nucleic acid/gene part and/or can with the nucleic acid of rna binding protein coding nucleic acid/gene recombination.Term with regard to functional variant " function " refers to such variant (i.e. part or hybridization sequences), its coding has RNA in conjunction with active polypeptide and preferably and additionally have at least one RRM, preferred 2 or 3 RRM and more preferably have one of following at least motif: have the motif of at least 75% sequence identity and/or the motif that has at least 50% sequence identity with motif II:RFDPFTGEPYKFDP with motif I:PYEAAVVALPVVVKERLVRILRLGIATRYD.Term " function " also can refer to the nucleic acid of or its homologue conjugated protein as the coding RNA of above this paper definition, makes plant have the growth characteristics of improvement when it is introduced in plant and expresses.

The nucleic acid of coding RBP1 polypeptide or its homologue can be any natural or synthetic nucleic acid.RBP1 polypeptide or its homologue as above this paper definition are by rbp1 nucleic acid/genes encoding.Therefore the term " rbp1 nucleic acid/gene " as definition here is coding as the RBP1 polypeptide of above this paper definition or any nucleic acid/gene of its homologue.The example of rbp1 nucleic acid comprises by those of arbitrary representative among SEQ ID NO:14, SEQ ID NO:16, SEQ ID NO:18, SEQ ID NO:20, SEQ ID NO:22, SEQID NO:24, SEQ ID NO:26, SEQ ID NO:28, SEQ ID NO:30, SEQ ID NO:32, SEQ ID NO:34 and the SEQ ID NO:36.Rbp1 nucleic acid/gene and functional variant thereof are applicable to implementing method of the present invention.The functional variant of rbp1 nucleic acid/gene comprise rbp1 nucleic acid/gene part and/or can with the nucleic acid of rbp1 nucleic acid/gene recombination.Term with regard to functional variant " function " refers to such variant (i.e. part or hybridization sequences), its encoded polypeptides has RNA in conjunction with active and have at least one RRM structural domain, preferred 2 RRM structural domains and more preferably have following two motifs: (i) KIFVGGL and (ii) RPRGFGF allow three aminoacid replacement and conservative arbitrarily change at the most in the motif.Term " function " also can refer to encode as the RBP1 polypeptide of above this paper definition or the nucleic acid of its homologue, makes plant have the growth characteristics of improvement when it is introduced in plant and expresses.

Term as defined herein partly refers to the protein-bonded dna fragmentation of coding RNA, be at least 180,300,500 or 700 according to the preference ordering length of nucleotides that increases, and this part encoded polypeptides has RNA in conjunction with active and at least 1 RRM, among preferred two or three RRM and motif I or the II at least one, preferred two are had both at the same time.For example, can prepare part by the rna binding protein coding nucleic acid is carried out one or more disappearances.Can use this part maybe it can be blended in other coding (or non-coding) sequence with isolating form and produce the some active protein of combination with (for example), one of them be that RNA is in conjunction with activity.When being blended in other encoding sequence, the polypeptide that the polypeptide that is produced by translation can partly be predicted greater than rna binding protein.Preferably, described funtion part is the part by the nucleic acid of arbitrary representative among SEQ ID NO:1, SEQ ID NO:3, SEQ ID NO:5, SEQ ID NO:7 and the SEQ ID NO:9.

Term with regard to rbp1 nucleic acid refers to that partly the dna fragmentation and the described fragment coding that comprise at least 80 Nucleotide have RNA in conjunction with active polypeptide, and has at least one RRM structural domain, preferred two RRM structural domains and further preferably have following two motifs: (i) KIFVGGL and (ii) RPRGFGF.For example, can prepare part by rbp1 nucleic acid is carried out one or more disappearances.Can use this part maybe it can be blended in other coding (or non-coding) sequence with isolating form and produce the protein that makes up various active with (for example), one of them be that RNA is in conjunction with activity.When being blended in other encoding sequence, can be by the polypeptide that translation produces greater than the rbp1 fragment polypeptide of prediction.Preferably, this funtion part is the part by the nucleic acid of arbitrary representative among SEQ ID NO:14, SEQ ID NO:16, SEQ IDNO:18, SEQ ID NO:20, SEQ ID NO:22, SEQ ID NO:24, SEQ ID NO:26, SEQ ID NO:28, SEQ ID NO:30, SEQ ID NO:32, SEQ ID NO:34 and the SEQID NO:36.

The rna binding protein variant of another type is under the stringent condition that reduces, preferably under stringent condition can with the nucleic acid as the nucleic acid/gene recombination of this paper rna binding protein defined previously coding, this hybridization sequences coding has RNA in conjunction with active polypeptide, and has at least 1 RRM, preferred two or three RRM, and at least one of motif I or II, preferred two.According to the preference ordering that increases, the length of nucleotides of this hybridization sequences is at least 180,300,500 or 700.Preferably, this hybridization sequences can with the nucleic acid hybridization by arbitrary representative among SEQ ID NO:1, SEQ ID NO:3, SEQ ID NO:5, SEQ ID NO:7 and the SEQ ID NO:9.

Similarly, the variant rbp1 of another type is under the stringent condition that reduces, preferably under stringent condition can with the nucleic acid as this paper rbp1 nucleic acid/gene recombination defined previously, this hybridization sequences coding has RNA in conjunction with active polypeptide, and has at least 1 RRM, preferred two RRM, and further preferably have following two motifs: (i) KIFVGGL and (ii) RPRGFGF.This hybridization sequences preferred length is at least 80 Nucleotide.Preferably, this hybridization sequences can with the nucleic acid hybridization by arbitrary representative among SEQ IDNO:14, SEQ ID NO:16, SEQ ID NO:18, SEQ ID NO:20, SEQ ID NO:22, SEQ ID NO:24, SEQ ID NO:26, SEQ ID NO:28, SEQ ID NO:30, SEQ IDNO:32, SEQ ID NO:34 and the SEQ ID NO:36.

Term " hybridization " as definition here refers to wherein basic homology complementary nucleotide sequence annealed process each other.Crossover process can take place in solution fully, and promptly complementary nucleic acid is all in solution.Crossover process also can be carried out in this case, and promptly one of complementary nucleic acid is fixed on the matrix, on magnetic bead, sepharose 4B or any other resin.In addition, crossover process also can so be carried out, promptly wherein one of complementary nucleic acid is fixed on solid support such as nitrocellulose or the nylon membrane, perhaps is fixed on (latter is referred to as nucleic acid array or microarray, or is referred to as nucleic acid chip) on for example siliceous glass support as photolithograph.For hybridization is taken place, make nucleic acid molecule thermally denature or chemical modification usually, so that two strands is unwind into two strands, and/or remove hairpin structure or other secondary structure in the single-chain nucleic acid.The severity of hybridization is subjected to such as condition effect such as temperature, salt concn, ionic strength and hybridization buffer compositions.Under the stringent condition that reduces, preferably under stringent condition, hybridize.The example of stringent condition shows in following table 3.Stringent condition is at least as strict as condition A-L; The stringent condition that reduces is at least as strict as condition M-R.

Table 3: the example of stringent condition

E	RNA:RNA	＞or equal 50	70 ℃.; 1 * SSC-or-50 ℃.; 1 * SSC, 50% methane amide	70℃.；0.3×SSC
					F	RNA:RNA	＜50	Tf *；1×SSC	Tf *；1×SSC
G	DNA:DNA	＞or equal 50	65 ℃.; 4 * SSC-or-45 ℃.; 4 * SSC, 50% methane amide	65℃.；1×SSC
					H	DNA:DNA	＜50	Th *；4°SSC	Th *；4×SSC
I	DNA:RNA	＞or equal 50	67 ℃.; 4 * SSC-or-45 ℃.; 4 * SSC, 50% methane amide	67℃.；1×SSC
					J	DNA:RNA	＜50	Tj *；4×SSC	Tj *；4×SSC
K	RNA:RNA	＞or equal 50	70 ℃.; 4 * SSC-or-40 ℃.; 6 * SSC, 50% methane amide	67℃.；1×SSC
					L	RNA:RNA	＜50	Tl *；2×SSC	Tl *；2×SSC
M	DNA:DNA	＞or equal 50	50 ℃.; 4 * SSC-or-40 ℃.; 6 * SSC, 50% methane amide	50℃.；2×SSC
					N	DNA:DNA	＜50	Tn *；6×SSC	Tn *；6×SSC
O	DNA:RNA	＞or equal 50	55 ℃.; 4 * SSC-or-42 ℃.; 6 * SSC, 50% methane amide	55×C.；2×SSC
					P	DNA:RNA	＜50	Tp *；6×SSC	Tp *；6×SSC
Q	RNA:RNA	＞or equal 50	60 ℃.; 4 * SSC-or-45 ℃.; 6 * SSC, 50% methane amide	60℃.；2×SSC
					R	RNA:RNA	＜50	Tr *；4×SSC	Tr *；4×SSC

" hybrid length " is the expection length of hybrid nucleic acid.When the nucleic acid hybridization of known array, can and differentiate that conservative region as herein described determines hybrid length by sequence alignment.

In hybridization and lavation buffer solution, can (1 * SSPE be 0.15M NaCl, 10mM NaH with SSPE ₂PO ₄With 1.25mM EDTA, pH7.4) replace SSC (1 * SSC is 0.15M NaCl and 15mM Trisodium Citrate); Hybridization was finished after scouring 15 minutes.Hybridization and washing can additionally comprise the fragmentation salmon sperm DNA, 0.5% trisodium phosphate of 5 * Denhardt ' s reagent, 0.5-1.0%SDS, 100 μ g/ml sex change and up to 50% methane amide. ^*Tb-Tr: for the heterozygote of expection length less than 50 base pairs, hybridization temperature should be than the melting temperature(Tm) T of heterozygote _mLow 5-10 ℃; Determine T according to following equation _mFor hybrid length less than 18 base pairs, T _m(℃)=2 * (A+T base number)+4 * (G+C base number).For hybrid length between 18 to 49 base pairs, Tm (℃)=81.5+16.6 * log ₁₀[Na ⁺]+0.41 * (%G+C)-(600/N), wherein N is the base number in the heterozygote, [Na ⁺] be the ([Na of 1 * SSC of Na ion concentration in the hybridization buffer ⁺]=0.165M).± the present invention is contained with peptide nucleic acid(PNA) (PNA) or nucleic acid arbitrary or a plurality of DNA of replacement that modifies or RNA hybridization mating partner.

Rna binding protein coding nucleic acid or its variant can be derived from any natural or artificial source.Can be from microbe-derived as bacterium, yeast or fungi, or from plant, algae or animal (comprising the people) source isolating nucleic acid/gene or its variant.Can modify this nucleic acid from its natural form composition and/or genome environment by meticulous manual operation.This nucleic acid preferred plant source, or from identical plant species (for example from will to exotic plant species wherein) or from different plant species.Described nucleic acid can preferably from tobacco (Nicotianae) section, more preferably separate from tobacco from dicotyledonous species.More preferably, represent by SEQ ID NO:1 from the isolating rna binding protein coding nucleic acid of tobacco, and rna binding protein aminoacid sequence such as SEQ ID NO:2 representative.

Rbp1 nucleic acid or its variant can be derived from any natural or artificial source.Can be from microbe-derived as bacterium, yeast or fungi, or from plant, algae or animal (comprising the people) source isolating nucleic acid/gene or its variant.Can modify this nucleic acid from its natural form composition and/or genome environment by meticulous manual operation.This nucleic acid preferred plant source, or from identical plant species (for example from will to exotic plant species wherein) or from different plant species.Described nucleic acid can preferably from cruciate flower (Brassicaceae) section, more preferably separate from Arabidopis thaliana from dicotyledonous species.More preferably, represent by SEQ ID NO:14 from the isolating rbp1 of Arabidopis thaliana, and RBP1 aminoacid sequence such as SEQ ID NO:15 representative.

Can increase the activity of rna binding protein or its homologue by introducing genetic modification (preferably at rna binding protein encoding gene seat).Similarly, can increase the activity of RBP1 polypeptide or its homologue by introducing genetic modification (preferably at the rbp1 locus).Here Ding Yi locus means the genome district, and it comprises goal gene and upstream of coding region or downstream 10KB.

For example, can introduce genetic modification by following any (or multiple) method: TDNA activation, TILLING, site-directed mutagenesis, homologous recombination or by in plant, introducing and express the nucleic acid of conjugated protein or its homologue of coding RNA or by in plant, introducing and express the nucleic acid of coding RBP1 polypeptide or its homologue.Introduce after the genetic modification, succeeded by the activity of selecting the active of rna binding protein increase or selecting the RBP1 polypeptide to increase, described active increase makes plant have the growth characteristics of improvement.

T-DNA activation tagging (Science (1992) 1350-1353 such as Hayashi) relates to the insertion of T-DNA, this T-DNA contains promotor (also can be translational enhancer or intron) usually, it is at genome district or the gene coding region upstream or the downstream 10KB of goal gene, and makes promotor can instruct target gene expression in configuration.Usually natural promoter is destroyed to the regulation and control of expression of target gene, and gene is by the promotor control of new introducing.Promotor generally is contained among the T-DNA.For example, infect by Agrobacterium (Agrobacterium) and this T-DNA inserted Plant Genome at random and cause near the gene overexpression that inserts the T-DNA.The transgenic plant that obtain are owing near the gene overexpression promotor of introducing shows the dominant phenotype.The promotor of introducing can be arbitrarily can be in desirable organism (is plant in this kind situation) instruct the promotor of genetic expression.For example, composing type, that organize preference, the cell type preference and promotor induction type are applicable to that all T-DNA activates.

Also can genetic modification be introduced rna binding protein encoding gene seat by TILLING (the local sudden change of the genome of targeted induction) technology.This is a kind of induced-mutation technique, is used to produce and/or identify, reach the variant that can present the active rna binding protein coding nucleic acid of rna binding protein (or rbp1 coding nucleic acid) of last separation mutagenesis.TILLING also allows to select to carry the plant of this kind mutation variants.These mutant even may present higher rna binding protein activity than its natural form gene.TILLING combines high-density mutagenesis and high-throughput screening method.The step that TILLING generally follows is: (a) EMS mutagenesis (Redei and Koncz, 1992; Feldmann etc., 1994; Lightner and Caspar, 1998); (b) DNA preparation and individual the merging; (c) pcr amplification in purpose zone; (d) sex change and annealing are to form heteroduplex; (e) DHPLC, wherein the heteroduplex that exists in the storehouse can detect extra peak on color atlas; (f) evaluation of mutated individual; (g) order-checking of sudden change PCR product.The method of TILLING is (McCallum NatBiotechnol.2000 Apr well-known in this area; 18 (4): 455-7, by Stemple summary, 2004 (TILLING-ahigh-throughput harvest for functional genomics.Nat Rev Genet.2004Feb; 5 (2): 145-50.)).

Site-directed mutagenesis can be used for producing the variant of rna binding protein coding nucleic acid or its part, described variant retentive activity, and promptly RNA is in conjunction with activity.Can finish site-directed mutagenesis by Several Methods, (current protocols in molecular biology.Wiley edits the method for the modal PCR of being based on Http:// www.4ulr.com/products/currentprotocols/index.html).Site-directed mutagenesis can be used for producing the variant of rna binding protein coding nucleic acid or its part, described variant retentive activity, and promptly RNA is in conjunction with activity.Similarly, site-directed mutagenesis can be used for producing the variant of RBP1 coding nucleic acid or its part, described variant retentive activity, and promptly RNA is in conjunction with activity.Site-directed mutagenesis also can be used for producing the variant of RBP1 coding nucleic acid or its part, described variant retentive activity, and promptly RNA is in conjunction with activity.

TDNA activation, TILLING and site-directed mutagenesis are to produce the example that new allelotrope and rna binding protein variant (this variant keeps the rna binding protein function) maybe can produce the technology of new allelotrope and rbp1 variant (this variant keeps the RBP1 function), so it is useful in the method for the invention.

Homologous recombination allows the appointment selected location in genome to introduce selected nucleic acid.Homologous recombination is the conventional standard technique of using in the bio-science, and it is used for low organism such as yeast or the mosses (as physcomitrella) of waiting.The method of in plant, carrying out homologous recombination not only in model plant, be described (Offringa etc., Extrachromosomal homologous recombination and genetargeting in plant cells after Agrobacterium-mediated transformation.1990 EMBO is Oct J.1990; 9 (10): 3077-84), and at crop plants, as obtaining describing (Terada R in the rice, Urawa H, Inagaki Y, Tsugane K, Iida S.Efficient genetargeting by homologous recombination in rice.Nat Biotechnol.2002.lida and Terada:A tale of two integrations, transgene and T-DNA:gene targetingby homologous recombination in rice.Curr Opin Biotechnol.2004 Apr; 15 (2): 132-8).The nucleic acid of institute's target (it may be that the rna binding protein coding nucleic acid that above defines or its variant or its may be rbp1 nucleic acid or its variants that above defines) does not need targeted rna binding-protein gene seat or target rbp1 locus, but can be introduced into for example zone of high expression level.The nucleic acid of institute's target can be the allelotrope of improvement, and it is used for replacing native gene or additionally is incorporated into native gene.

According to the preferred embodiments of the invention, by in plant, introducing and expressing coding RNA and can improve the growth characteristics of plant in conjunction with the nucleic acid of polypeptide or its homologue, described polypeptide or its homologue have RNA and discern motifs (RRM) in conjunction with active and 2 or 3 RNA, and it comprises with motif I:PYEAAVVALPVVVKERLVRILRLGIATRYD and has the motif of at least 75% sequence identity and/or the motif that has at least 50% sequence identity with motif II:RFDPFTGEPYKFDP.

The preferred method of introducing genetic modification (it does not need in the rna binding protein locus in this case) is the nucleic acid of introducing and expressing conjugated protein or its homologue of the coding RNA of definition as mentioned in plant.

Another preferred embodiment according to the present invention can be by introducing and express the nucleic acid improvement plant growth characteristics of coding RBP1 polypeptide or its homologue in plant.

A preferred method introducing genetic modification (it does not need in the rbp1 locus in this case) is to introduce and express the coding RBP1 polypeptide of definition as mentioned or the nucleic acid of its homologue in plant.RBP1 polypeptide or its homologue as above mentioned have: (a) RNA is in conjunction with activity; (b) two RRM structural domains; (c) following two motifs: (i) KIFVGGL and (ii) RPRGFGF allow three aminoacid replacement and conservative arbitrarily change the at the most in the motif; (d) amino acid according to preference ordering that increases and SEQID NO:15 representative has at least 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95% sequence identity.

Proteinic " homologue " contains peptide, oligopeptides, polypeptide, protein and enzyme, it has amino acid whose replacement, disappearance and/or insertion with respect to the unmodified protein matter of being discussed, and have to its derived from similar biologic activity and the functionally active of unmodified protein matter.In order to produce such homologue, proteinic amino acid can be replaced by other amino acid with similar quality (as similar hydrophobicity, wetting ability, antigenicity, form or break the tendency of αLuo Xuanjiegou or β laminated structure).Conservative replacement table is well-known in this area (for example seeing Creighton (1984) Proteins.W.H.Freeman and Company).Following table provides the example that conserved amino acid replaces.

Table 4: the example that conserved amino acid replaces

Residue	The conservative replacement	Residue	The conservative replacement
				Ala	Ser	Leu	Ile、Val
Arg	Lys	Lys	Arg、Gln
				Asn	Gln、His	Met	Leu、Ile
Asp	Glu	Phe	Met、Leu、Tyr
				Gln	Asn	Ser	Thr、Gly
Cys	Ser	Thr	Ser、Val
				Glu	Asp	Trp	Tyr
Gly	Pro	Tyr	Trp、Phe
				His	Asn、Gln	Val	Ile、Leu
Ile	Leu、Val

The homologue of two kinds of specific forms also contained in term " homologue ", and it comprises lineal homologous sequence and collateral line homologous sequence, and it contains the evolution notion that is used to describe the gene ancestral relationship.Term " collateral line homology " relates to the collateral line gene that the gene replication in species gene group inside produces.Term " lineal homology " relates to because species form the homologous gene in the different organisms that produce.

For example can easily find for example lineal homologue in the monocotyledons species by carrying out so-called mutual (reciprocal) blast search.This can by use the sequence of studying (for example SEQID NO:1 or 2 or SEQ ID NO:14 or 15) realize by a blast at any sequence library (such as the public obtainable ncbi database that can find at http://www.ncbi.nlm.nih.gov).For example, if the lineal homologue in the searching rice should carry out in 28,469 full length cDNA clones at rice (Oryza sativa) Nipponbare that can obtain from NCBI the blast that studies sequence.When Nucleotide begins, using BLASTn or tBLASTX, maybe when when protein begins, using BLASTP or TBLASTN with standard default with standard default.Blast result can filter.Then use the full length sequence among filtering result or the unfiltered result to carry out reverse blast (secondary blast) at the sequence of research sequence source organism certainly.The result who compares the first time and second time blast then.When described secondary blast result provides when having the highest similarity with rna binding protein coding nucleic acid or rna binding protein polypeptide, be and find lineal homologue, for example,, find the collateral line homologue so if one of organism is a tobacco.For RBP1, when described secondary blast result provides when having the highest similarity with rbp1 nucleic acid or RBP1 polypeptide, be and find lineal homologue, for example,, find the collateral line homologue so if one of organism is an Arabidopis thaliana.Can use Clustal W under the situation of extended familys, it is visual to help cluster succeeded by contiguous threaded tree.

Homologue may be the form of proteinic " replacement variant ", promptly has at least a residue to be removed in aminoacid sequence, and inserts different residues in this position.Aminoacid replacement is the replacement of single residue normally, but determines it also may is that cluster replaces by the functional limitations factor of polypeptide; Insert usually at about 1 to 10 amino-acid residue order of magnitude.Preferably, aminoacid replacement comprises conservative aminoacid replacement.

Homologue also can be the form of proteinic " insertion variant ", promptly introduces one or more amino-acid residues in proteinic predetermined position.Insertion can comprise the fusion of aminoterminal and/or carboxyl terminal, and the inner insertion of single or multiple amino acid whose sequence.The general order of magnitude will be less than the fusion of amino or carboxyl terminal in the insertion of the aminoacid sequence inside of about 1 to 10 residue.The example of amino or carboxyl terminal fused protein or peptide is included in binding domains or activation structure territory, bacteriophage coat protein matter, (Histidine) 6-label, glutathione S-transferase label, a-protein, maltose binding protein, Tetrahydrofolate dehydrogenase, Tag100 epi-position, c-myc epi-position, FLAG  epi-position, lacZ, CMP (calmodulin binding peptide), HA epi-position, protein C epi-position and the VSV epi-position of the activating transcription factor of using in the yeast two-hybrid system.

The homologue of protein " disappearance variant " form is characterised in that removes one or more amino acid from protein.

Can use well-known peptide synthetic technology in this area,, or prepare proteinic amino acid variant by recombinant DNA processing ease ground as the solid phase method of peptide synthesis etc.The dna sequence dna working method that is used to produce proteinic replacement, insertion or lack variant is well-known in this area.For example, producing the technology that replaces sudden change in the DNA predetermined position is that those skilled in that art are well-known, comprise M13 mutagenesis, T7-Gen vitro mutagenesis (USB, Cleveland, OH), QuickChange site-directed mutagenesis (Stratagene, San Diego, CA), site-directed mutagenesis or other site-directed mutagenesis method of PCR mediation.

Rna binding protein or its homologue can be that derivative or RBP1 polypeptide or its homologue can be derivatives." derivative " comprises peptide, oligopeptides, polypeptide, protein and enzyme, with the natural generation form of protein (for example SEQ ID NO:2 representative or under the RBP1 situation SEQ ID NO:15 representative) aminoacid sequence compares, it can comprise replacement, disappearance or the interpolation natural and amino-acid residue that non-natural produces.Proteinic " derivative " contains peptide, oligopeptides, polypeptide, protein and enzyme, compares with the aminoacid sequence of the natural generation form of polypeptide, and it can comprise the amino-acid residue that natural that change, glycosylated, acylations or non-natural produces.The one or more non-aminoacid replacement that derivative can also comprise with respect to its aminoacid sequence that is derived from (for example is incorporated into reporter molecules or other part of aminoacid sequence covalently or non-covalently, and the amino-acid residue that non-natural produces for the proteinic aminoacid sequence of natural generation the reporter molecules that helps detecting as bonded with it).

Rna binding protein or its homologue can be by the alternative splicing variant codings of rna binding protein nucleic acid/gene.RBP1 polypeptide or its homologue can be by the alternative splicing variant codings of rbp1 nucleic acid/gene.The intron of selecting contained wherein in term used herein " alternative splicing variant " and/or exon is cut, replace or the variant of the nucleotide sequence that adds.Such variant has kept proteinic biological activity, and this can be by the functional fragment realization of retaining protein selectively.Such splice variant can be natural or artificial.The method that produces this splice variant is well-known in the art.Preferred splice variant is the splice variant of the nucleic acid of SEQ ID NO:1, SEQ ID NO:3, SEQ IDNO:5, SEQ ID NO:7 and SEQ ID NO:9 representative.Preferred splice variant encoded polypeptides has kept RNA in conjunction with active and have at least one RRM, at least one among preferred 2 or 3 RRM and motif I or the II, and preferably both have concurrently.The splice variant of preferred RBP1 is the splice variant by the nucleic acid of SEQ ID NO:15, SEQ ID NO:17, SEQ ID NO:19, SEQ ID NO:21, SEQ ID NO:23, SEQ ID NO:25 or SEQ ID NO:27, SEQID NO:29, SEQ ID NO:31, SEQ ID NO:33, SEQ ID NO:35 and SEQ IDNO:37 representative.How preferred splice variant encoded polypeptides has kept RNA in conjunction with active and have preferred two RRM structural domains and further preferred following two motifs: (i) KIFVGGL and (ii) RPRGFGF, the change that allows three aminoacid replacement at the most in the motif and guard arbitrarily.

Homologue can also be conjugated protein by coding RNA or the allele variant coding of the nucleic acid of its homologue, the allele variant of the nucleic acid of arbitrary representative among preferred SEQ ID NO:1, SEQ ID NO:3, SEQ ID NO:5, SEQ ID NO:7 and the SEQ ID NO:9.Further preferably have RNA in conjunction with active and have at least one RRM by the allele variant encoded polypeptides, preferred 2 or 3 RRM and motif I or II genuine at least one, preferably both have concurrently.Homologue can also be by the allele variant coding of the nucleic acid of coding RBP1 polypeptide or its homologue, preferably by the allele variant of the nucleic acid of SEQ ID NO:15, SEQ ID NO:17, SEQ ID NO:19, SEQ ID NO:21, SEQID NO:23, SEQ ID NO:25 or SEQ ID NO:27, SEQ ID NO:29, SEQ IDNO:31, SEQ ID NO:33, SEQ ID NO:35 and SEQ ID NO:37 representative.Further preferably has RNA in conjunction with active and preferred 2 RRM structural domains and following two motifs: (i) KIFVGGL and (ii) RPRGFGF, permission three aminoacid replacement and the change of guarding arbitrarily at the most in the motif by the allele variant encoded polypeptides.The natural existence of allele variant, and these natural allelic purposes are contained in the method for the present invention.Allele variant comprises single nucleotide polymorphism (SNP), and small-sized insertion/deletion polymorphism (INDEL).The size of INDEL is usually less than 100bp.SNP and INDEL form one group of maximum sequence variants in the naturally occurring polymorphism strain of most of organisms.

According to a preferred aspect of the present invention, the expression of rna binding protein coding nucleic acid or its variant increases or increases.According to a preferred aspect of the present invention, the expression of rbp1 nucleic acid or its variant increases or increases.The gene that to improve or increase or the method for gene product expression have sufficient record in this area, it comprises, for example by the use of crossing expression, transcriptional enhancer or translational enhancer of suitable promoters driven.The appropriate location (generally being the upstream) that the isolating nucleic acid that is used as promotor or enhancer element can be introduced non-allos form polynucleotide, thereby the expression of rise rna binding protein coding nucleic acid or its variant.For example, can and/or replace by sudden change, disappearance, body changes endogenesis promoter interiorly and (sees Kmiec, U.S. Patent number 5,565,350; Zarling etc., PCT/US93/03868), perhaps with isolating promotor in the suitable direction of gene of the present invention with in apart from the introduced plant cell, thereby the expression of controlling gene.

If wish expression of polypeptides, to comprise polyadenylation region at the 3 '-end in peptide coding zone usually.Polyadenylation region can be derived from natural gene, multiple other plant gene or T-DNA.For example, 3 ' of adding end sequence can be derived from nopaline synthase or octopine synthase gene or alternatively from other plant gene or less preferred from any other eukaryotic gene.

Also can in 5 ' non-translational region of part encoding sequence or encoding sequence, add intron sequences, be increased in the quantity of the ripe courier of cumulative in the kytoplasm.Show, but the intron of the montage that comprises in the transcription unit of plant and animal expression construct all can increase genetic expression up to 1000 times at mRNA and protein level simultaneously, Buchman and Berg, Mol.Cell biol.8:4395-4405 (1988); Callis etc., Genes Dev.1:1183-1200 (1987).Usually when this intron be placed on transcription unit 5 ' terminal near the time, its effect that improves genetic expression reaches maximum.Corn intron A dh1-S introne 1,2 and 6, the use of Bronze-1 intron is well known in the art.Usually see The Maize Handbook, the 116th chapter, Freeling and Walbot edit, Springer, N.Y. (1994).

The present invention also provides genetic constructs and carrier, is used for the introducing and/or the expression of the nucleotide sequence of the inventive method with promotion.

Therefore, the gene construct that provides comprises:

(i) rna binding protein coding nucleic acid or its variant;

The control sequence that (ii) one or more nucleotide sequences that can drive (i) are expressed; With optional

(iii) transcription termination sequence.

The gene construct that also provides comprises:

(i) rbp1 nucleic acid or its variant;

The control sequence that (ii) one or more nucleotide sequences that can drive (i) are expressed; With optional

(iii) transcription termination sequence.

Can use the well-known recombinant DNA technology of those skilled in the art to make up the construct that is used for the inventive method.Gene construct can be inserted in (commercially available) carrier, described carrier is suitable for transforming and enters plant and be suitable for expressing goal gene in cell transformed.

The carrier that use comprises aim sequence (being rna binding protein coding nucleic acid or its variant or rbp1 nucleic acid or its variant) transforms plant.Aim sequence effectively is connected in one or more control sequences (being connected in promotor at least).Term " controlling element ", " control sequence " and " promotor " all tradable in this article use, and general reference can influence the regulation and control nucleotide sequence that its catenation sequence is expressed in context.Above-mentioned term is contained and is derived from typical eukaryotic gene group gene and (comprises the TATA box that has or do not have CCAAT box sequence, it is essential for accurate transcription initiation) transcription regulating nucleotide sequence, and other controlling element (being upstream activating sequence, enhanser and silencer), it is by replying growth stimulation and/or outside stimulus or changing genetic expression in tissue-specific mode.This term has also comprised the transcription regulating nucleotide sequence of classical prokaryotic gene, and it can comprise-35 box sequences and/or-10 box transcription regulating nucleotide sequences in the case.Synthetic fusion molecule or derivative also contained in term " controlling element ", and it gives, activates or improve the expression of cell, tissue or organ amplifying nucleic acid molecule.Term used herein " effectively connects " and refers to the functional connection between promoter sequence and goal gene, thereby makes transcribing of the promoter sequence initial goal gene of energy.

Advantageously, can use the expression of the promoters driven nucleotide sequence of any type.Promotor can be an inducible promoter, promptly replys the stimulation of growth, chemistry, environment or physics, has the transcription initiation of inductive or increase.The example of inducible promoter is a stress induced promoter, promptly is exposed to multiple stress conditions activated of following time promotor when plant.In addition or alternative, described promotor can be to organize the promotor of preference, promptly can organize at some, as preferentially initial promotor of transcribing in tissues such as leaf, root, seed.

Preferably, rna binding protein coding nucleic acid or its variant effectively are connected in the promotor of seed preference.The promotor of seed preference is a preference, but need not to drive in seed tissue expression promoter single-mindedly.Preferred seed tissue is an endosperm.Preferred promotor is prolamine (prolamin) promotor, for example from the prolamine promotor (SEQ ID NO:11) of rice.Should be clear and definite be the rna binding protein coding nucleic acid that application of the present invention is not limited to SEQ ID NO:1 representative, the expression of rna binding protein coding nucleic acid when also being not limited to by the prolamine promoters driven.

Preferably, rbp1 nucleic acid or its variant effectively be connected in can be in bud the promotor of preference ground express nucleic acid.Preferably, can in bud, the promotor of preference ground express nucleic acid have and the similar express spectra of β expansion protein promoter, for example shown in Figure 5.Most preferably, can be in bud the promotor of preference ground express nucleic acid from the β expansion protein promoter (SEQ ID NO:38) of rice.Should be clear and definite be the rbp1 nucleic acid that application of the present invention is not limited to SEQ ID NO:14 representative, rbp1 expression of nucleic acids when also being not limited to drive by β expansion protein promoter.

Choose wantonly, can also in the construct of introduced plant, use one or more terminator sequences.Control sequence contained in term " terminator ", and this dna sequence dna is positioned at transcription unit's end, 3 ' processing of transmission signal initiation primary transcript and polyadenylation and the termination of transcribing.Other controlling element can comprise the enhanser of transcribing and translating.Those skilled in the art will know and be suitable for carrying out terminator of the present invention and enhancer sequence.

Genetic constructs of the present invention also is included in the starting point of keeping in the particular cell types and/or duplicating required replication sequence.Example be when needs with genetic constructs as additive type genetic elements (as a plasmid or a glutinous grain molecule) when in bacterial cell, keeping.Preferred replication orgin includes, but is not limited to f1-ori and colE1.

Genetic constructs can randomly comprise selectable marker gene.As used herein, term " selectable marker gene " comprises any gene of giving cell phenotype, and the expression of this gene in cell helps identifying and/or selecting use nucleic acid construct transfection of the present invention or cell transformed.Suitable mark can be selected from the mark that has microbiotic or Herbicid resistant, and it is introduced new metabolic characteristic or allows visual selection.The example of selectable marker gene comprises the gene (for example npt II of phosphorylation Xin Meisu and kantlex, or the hpt of phosphorylation Totomycin) of giving antibiotics resistance, and (for example bar provides the resistance to Basta to the gene of conferring herbicide resistance; AroA or gox provide the resistance to glyphosate), or the gene (using the manA of seminose as sole carbon source as allowing plant) of metabolic characteristic is provided.Visual mark cause form color (β-glucuronidase for example, GUS), luminous (for example luciferase) or fluorescence (green fluorescent protein GFP and derivative thereof).

The present invention is also contained can be by the plant of the inventive method acquisition.Therefore the present invention provides and can introduce rna binding protein coding nucleic acid or its variant or rbp1 nucleic acid or its variant in this plant by the plant of the inventive method acquisition.

The present invention also is provided for producing the method for the transgenic plant with improvement growth characteristics, and it is included in the plant and introduces and the conjugated protein coding nucleic acid of expressed rna or its variant.

More specifically, the invention provides the method that is used to produce the transgenic plant with improvement growth characteristics, this method comprises:

(i) in plant or vegetable cell, introduce rna binding protein coding nucleic acid or its variant; With

(ii) culturing plants cell under the condition that promotes plant-growth and growth.

The present invention also is provided for producing the method for the transgenic plant with improvement growth characteristics, and it is included in introduces and express rbp1 nucleic acid or its variant in the plant.

More specifically, the invention provides the method that is used to produce the transgenic plant with improvement growth characteristics, this method comprises:

(iii) in plant or vegetable cell, introduce rbp1 nucleic acid or its variant; With

(iv) culturing plants cell under the condition that promotes plant-growth and growth.

Can be with direct introduced plant cell of nucleic acid or plant itself (comprising any other parts of introducing tissue, organ or plant).According to preferred feature of the present invention, preferably by transforming with the nucleic acid introduced plant.

Here indication term " conversion " is contained exogenous polynucleotide is shifted into host cell, and does not consider to shift used method.Can use genetic constructs of the present invention to transform by organ takes place or the embryogenetic plant tissue that carries out clonal expansion that can continue and from its regeneration whole plants.Concrete tissue is selected and will be determined by the clonal expansion system of available and the concrete species of conversion that are suitable for most and change.Exemplary target tissue comprises the meristematic tissue (for example apical meristem, axillalry bud and root meristematic tissue) and the inductive meristematic tissue (for example cotyledon meristematic tissue and hypocotyl meristematic tissue) of leaf dish, pollen, embryo, cotyledon, hypocotyl, megagamete, callus, existence.Can be with polynucleotide instantaneous or stably introduce host cell, and can, for example keep nonconformable state as plasmid.Alternatively, it can be integrated into host genome.The transformed plant cells that obtains can be then used in well known to a person skilled in the art the mode plant transformed of regenerating.

The conversion of plant species is a kind of quite conventional technology at present.Advantageously, can use arbitrary in several method for transformation that goal gene is introduced suitable ancester cell.Method for transformation comprises with the chemical of liposome, electroporation, the picked-up of enhancing dissociative DNA, directly bombards, transforms and microprojection (microprojection) with virus or pollen to plant injection DNA, particle gun.Method can be selected from calcium/polyoxyethylene glycol method of being used for protoplastis (Krens, F.A. etc., 1882, Nature 296,72-74; Negrutiu I. etc., June 1987, Plant Mol.Biol.8,363-373), electroporation (the Shillito R.D. etc. of protoplastis, 1985 Bio/Technol 3,1099-1102), microinjection (Crossway A. etc., 1986, the Mol.Gen Genet 202 of vegetable material, 179-185), particle bombardment (the Klein T.M. etc. of DNA or RNA bag quilt, 1987, Nature 327,70), (nonconformable) virus infection or the like.The preferred protein-bonded transgenosis rice of conversion generation expressed rna of using any rice method for transformation of knowing via the Agrobacterium mediation, the described rice method for transformation method of describing in the document of how playing for example in office: disclosed European patent application EP 1198985 A1, Aldemita and Hodges (Planta, 199,612-617,1996); Chan etc. (Plant Mol.Biol.22 (3) 491-506,1993), Hiei etc. (Plant is (2) 271-282 J.6,1994), its disclosure is introduced into as a reference as the full content of its statement.Transform as for corn, preferable methods is as at (Nat.Biotechnol.1996 such as Ishida, 14 (6): 745-50) or Frame etc. (Plant Physiol.2002,129 (1): 13-22) described, its disclosure is introduced into as a reference as the full content of its statement.

Usually after transforming, select the vegetable cell or the groups of cells that there are one or more marks, described mark is by the expressive gene of plant coding that moves with the goal gene corotation, and the material regeneration with transforming that continues becomes whole plants.

After DNA transfer and the regeneration, can assess the conversion plant of supposition, for example form with existence, copy number and/or the genome of Southern analysis purposes gene.Alternative or extra, available Northern and/or Western analyze the expression level of the new DNA of introducing of monitoring, and these two kinds of technology all are that those skilled in the art are well-known.

The conversion plant that produces can breed in several ways, as the breeding technique with clonal propagation or classics.For example, the first-generation (or T1) but the s-generation (or T2) transformant that the plant transformed selfing obtains isozygotying, further by classical breeding technique breeding T2 plant.

The inverting biological body that produces can have various ways.For example, they can be the mosaics of transformant and no transformed cells; Clone's transformant (for example through transforming all cells that contains expression cassette); Transform and the graft of unconverted tissue (for example in plant, the conversion rhizome of grafting to the unconverted scion).

The present invention obviously extends to any vegetable cell or the plant that is produced by methods described herein, and the part of all plants and its vegetative propagule.The former generation conversion that produced by any aforesaid method or the offspring of cells transfected, tissue, organ or whole plants are also contained in the present invention, and unique requirement of described offspring is that the parent who produces with the inventive method presents same genotype and/or phenotypic characteristic.The present invention also comprises the host cell that contains separative rna binding protein nucleic acid or its variant.The preferred host cell of the present invention is a vegetable cell.The present invention also extends to the part that plant can be gathered in the crops, and for example, but is not limited to seed, leaf, fruit, flower, stem culture, rhizome, stem tuber and bulb.

The purposes of rna binding protein nucleic acid or its variant and the purposes of rna binding protein or its homologue are also contained in the present invention.

A kind of such purposes relates to the growth characteristics that improve plant, particularly improves productive rate, especially the seed productive rate.The seed productive rate can comprise following one or more: (full) seed number of increase, the seed weight of increase, harvest index of increase or the like.

Can use rna binding protein coding nucleic acid or its variant in the procedure of breeding, perhaps rna binding protein or its homologue are wherein identified the dna marker that can be connected in rna binding protein encoding gene or its variant hereditarily.Can use rna binding protein coding nucleic acid or its variant, perhaps rna binding protein or its homologue define molecule marker.Then this DNA or protein labeling can be used for the procedure of breeding, have the plant of the growth characteristics of change with selection.For example, rna binding protein encoding gene or its variant can be the nucleic acid by any one representative among SEQ ID NO:1, SEQ ID NO:3, SEQID NO:5, SEQ ID NO:7 and the SEQ ID NO:9.

The allele variant of rna binding protein encoding gene/nucleic acid also can be used for the auxiliary procedure of breeding of mark.Such procedure of breeding needs to use sometimes, and for example EMS mutagenesis is introduced allele variant by the mutagenic treatment of plant; Alternative, this program can begin with the allele variant of collecting what is called " natural " origin that is not intended to generation.Identify allele variant by for example PCR then.Be to select step in order to select the better allele variant of described sequence subsequently, described allele variant produces the growth characteristics of improvement in plant.Generally the growth behavior that contains the different allele variant plants of studying sequence to some extent by monitoring is selected, for example arbitrary different allele variants among SEQ ID NO:1, SEQ IDNO:3, SEQ ID NO:5, SEQ ID NO:7 and the SEQ ID NO:9.Can in greenhouse or field, monitor growth behavior.More optional step comprise, will contain plant and another plant hybridization of better allele variant through evaluation.For example, can make the combination that produces the purpose phenotypic characteristic in this way.

Rna binding protein coding nucleic acid or its variant can also be as probes, be used for it is carried out the mapping of heredity and physics for the gene of a gene part, and the chain proterties mark of those genes of being linked to each other of conduct.These information can be used in plant breeding, to obtain having the strain of desired phenotype.This class of rna binding protein coding nucleic acid or its variant is used the one section nucleotide sequence that only needs at least 15 Nucleotide long.Rna binding protein coding nucleic acid or its variant also can be used as restriction fragment length polymorphism (RFLP) mark.Available rna binding protein coding nucleic acid or its variant are surveyed the Southern trace (Maniatis) of the plant genome DNA of restriction digest.The program that uses a computer subsequently such as MapMaker (Lander etc., 1987) carry out genetic analysis to the banding pattern that produces, to make up genetic map.In addition, can use nucleic acid to survey the Southern trace in the genomic dna that the restriction enzyme that contains one group of individuality is handled, described one group of individuality is the parent of the clear and definite genetic cross of representative and one group of individuality of filial generation.The separation of dna polymorphism is recorded and is used for calculating formerly with the genetic map rna binding protein coding nucleic acid of this colony's acquisition or the position (Botstein etc. (1980) Am.J.Hum.Genet.32:314-331) of its variant.

The derive generation and the purposes of probe of the plant gene that uses in genetic mapping is described among Bematzky and Tanksley (1986) the Plant Mol.Biol.Reporter 4:37-41.Described in a lot of publications with aforesaid method or its flexible form specific cDNA clone was carried out genetic mapping.For example, can use F2 hybridization colony, backcross population, panmictic population, the homogenic system of close relative and the individual mapping of other group.These methods are that those skilled in the art are well-known.

Nucleic acid probe also can be used for physical mapping and (promptly settle sequence on physical map; See Hoheisel etc.: Non-mammalian Genomic Analysis:A Practical Guide, Academicpress 1996, the 319-346 pages or leaves, and the reference of wherein quoting).

In another embodiment, nucleic acid probe can be used for direct fluorescence in situ hybridization (FISH) mapping (Trask (1991) Trends Genet.7:149-154).Although the method inclination of FISH mapping at present uses big clone (several to a hundreds of KB; See (1995) Genome Res.5:13-20 such as Laan), but the raising of susceptibility allows to use short probe in the FISH mapping.

Can use described nucleic acid to be used for the multiple method of heredity and physical mapping based on nucleic acid amplification.Example comprises the polymorphism (CAPS of allele specific amplification (Kazazian (1989) J.Lab.Clin.Med11:95-96), pcr amplified fragment; Sheffield etc. (1993) Genomics16:325-332), allele-specific connects (Landegren etc. (1988) Science241:1077-1080), Nucleotide extension (Sokolov (1990) Nucleic Acid Res.18:3671), radiation hybridization mapping (Walter etc. (1997) Nat.Genet.7:22-28) and Happy mapping (Dear and Cook (1989) Nucleic Acid Res.17:6795-6807).For implementing these methods, it is right to use the nucleotide sequence design and produce the primer that is used for amplified reaction or primer extension reaction.This class primer design is that those skilled in that art are well-known.Use the method for the genetic mapping of PCR-based, may need to identify the difference of leap corresponding to dna sequence dna between the parent of current nucleotide sequence zone mapping.Yet this is dispensable usually to drawing method.

Also find rna binding protein coding nucleic acid or its variant, perhaps rna binding protein or its homologue are as the purposes of growth regulator.Because it is useful that these molecules have been presented in the improvement plant growth characteristics, they also are useful growth regulators, as weedicide or growth stimulator.Therefore, the invention provides the composition as growth regulator, it comprises rna binding protein coding nucleic acid/gene or its variant or rna binding protein or its homologue and appropriate carriers, thinner or vehicle.

The purposes of rbp1 nucleic acid or its variant and the purposes of RBP1 polypeptide or its homologue are also contained in the present invention.

A kind of such purposes relates to the growth characteristics that improve plant, particularly improves productive rate, especially the seed productive rate.The seed productive rate can comprise following one or more: (full) seed number of increase, the seed weight of increase, harvest index of increase or the like.

Can use rbp1 nucleic acid or its variant in the procedure of breeding, perhaps RBP1 polypeptide or its homologue are wherein identified the dna marker that can be connected in rbp1 gene or its variant hereditarily.Can use rbp1 nucleic acid or its variant, perhaps RBP1 or its homologue limit molecule marker.Then this DNA or protein labeling can be used for the procedure of breeding, have the plant of the growth characteristics of change with selection.For example, rbp1 gene or its variant can be the nucleic acid by arbitrary representative among SEQ ID NO:14, SEQ IDNO:16, SEQ ID NO:18, SEQ ID NO:20, SEQ ID NO:22, SEQ ID NO:24, SEQ ID NO:26, SEQ ID NO:28, SEQ ID NO:30, SEQ ID NO:32, SEQ IDNO:34 and the SEQ ID NO:36.

The allele variant of rbp1 also can be used for the auxiliary procedure of breeding of mark.Such procedure of breeding needs to use sometimes, and for example EMS mutagenesis is introduced allele variant by the mutagenic treatment of plant; Alternative, this program can begin with the allele variant of collecting what is called " natural " origin that is not intended to generation.Identify allele variant by for example PCR then.Be to select step in order to select the better allele variant of described sequence subsequently, described allele variant produces the growth characteristics of improvement in plant.Generally the growth behavior that contains the different allele variant plants of studying sequence to some extent by monitoring is selected, for example the arbitrary different allele variants among SEQ ID NO:14, SEQ ID NO:16, SEQ ID NO:18, SEQ ID NO:20, SEQ ID NO:22, SEQ ID NO:24, SEQ ID NO:26, SEQ IDNO:28, SEQ ID NO:30, SEQ ID NO:32, SEQ ID NO:34 and the SEQ ID NO:36.Can in greenhouse or field, monitor growth behavior.More optional step comprise, and will contain plant and another plant hybridization of better allele variant through evaluation.For example, can make the combination that produces the purpose phenotypic characteristic in this way.

Rbp1 nucleic acid or its variant can also be as probes, be used for it is carried out the mapping of heredity and physics for the gene of a gene part, and the chain proterties mark of those genes of being linked to each other of conduct.These information can be used in plant breeding, to obtain having the strain of desired phenotype.This class of rbp1 nucleic acid or its variant is used the one section nucleotide sequence that only needs at least 15 Nucleotide long.Rbp1 nucleic acid or its variant also can be used as restriction fragment length polymorphism (RFLP) mark.Available rbp1 nucleic acid or its variant are surveyed the Southern trace (Maniatis) of the plant genome DNA of restriction digest.The program that uses a computer subsequently such as MapMaker (Lander etc., 1987) carry out genetic analysis to the banding pattern that produces, to make up genetic map.In addition, can use nucleic acid to survey the Southern trace in the genomic dna that the restriction enzyme that contains one group of individuality is handled, described one group of individuality is the parent of the clear and definite genetic cross of representative and one group of individuality of filial generation.The separation of record dna polymorphism also is used for calculating formerly with the genetic map rbp1 nucleic acid of this colony's acquisition or the position (Botstein etc. (1980) Am.J.Hum.Genet.32:314-331) of its variant.

The derive generation and the purposes of probe of the plant gene that uses in genetic mapping is described among Bematzky and Tanksley (1986) the Plant Mol.Biol.Reporter 4:37-41.Described in a lot of publications with aforesaid method or its flexible form specific cDNA clone was carried out genetic mapping.For example, can use F2 hybridization colony, backcross population, panmictic population, the homogenic system of close relative and the mapping of other group of individuals.These class methods are that those skilled in the art are well-known.

Nucleic acid probe also can be used for physical mapping and (promptly settle sequence on physical map; See Hoheisel etc.: Non-mammalian Genomic Analysis:A Practical Guide, Academicpress 1996, the 319-346 pages or leaves, and the reference of wherein quoting).

In another embodiment, nucleic acid probe can be used for direct fluorescence in situ hybridization (FISH) mapping (Trask (1991) Trends Genet.7:149-154).Although the method inclination of FISH mapping at present uses big clone (several to a hundreds of KB; See (1995) Genome Res.5:13-20 such as Laan), but the raising of susceptibility allows to use short probe in the FISH mapping.

The multiple method based on nucleic acid amplification that is used for heredity and physical mapping can use described nucleic acid to carry out.Example comprises the polymorphism (CAPS of allele specific amplification (Kazazian (1989) J.Lab.Clin.Med11:95-96), pcr amplified fragment; Sheffield etc. (1993) Genomics16:325-332), allele-specific connects (Landegren etc. (1988) Science241:1077-1080), Nucleotide extension (Sokolov (1990) Nucleic Acid Res.18:3671), radiation hybridization mapping (Walter etc. (1997) Nat.Genet.7:22-28) and Happy mapping (Dear and Cook (1989) Nucleic Acid Res.17:6795-6807).For implementing these methods, it is right to use the nucleotide sequence design and produce the primer that is used for amplified reaction or primer extension reaction.This class primer design is that those skilled in that art are well-known.Use the method for the genetic mapping of PCR-based, may need to identify the difference of leap corresponding to dna sequence dna between the parent of current nucleotide sequence zone mapping.Yet this is dispensable usually to drawing method.

Also find rbp1 nucleic acid or its variant, perhaps RBP1 polypeptide or its homologue are as the purposes of growth regulator.Because it is useful that these molecules have been presented in the improvement plant growth characteristics, they also are useful growth regulators, as weedicide or growth stimulator.Therefore, the invention provides the composition as growth regulator, it comprises rbp1 or its variant or RBP1 polypeptide or its homologue and appropriate carriers, thinner or vehicle.

The method according to this invention is had the plant of improvement growth characteristics as previously mentioned.These favourable growth characteristics can also make up other favourable economically proterties, as the proterties of further raising output, to multiple tolerance of coercing, change the proterties of various structural attitudes and/or biochemistry and/or physiologic character.

Accompanying drawing is described.

With reference to following accompanying drawing the present invention is described, wherein:

Fig. 1 shows that the protein-bonded CLUSTAL multiple ratio of plant RNA is right.Motif I and II are added frame (BAC83046 lacks M2), and the RRM structural domain underlines.

Fig. 2 shows binary vector, and it is used for the tobacco rna binding protein under the control of rice expression prolamine promotor.

Fig. 3 shows that the multiple ratio of plant RBP1 polypeptide is right.Gene pool protein or its coding nucleic acid have been shown.At refers to Arabidopis thaliana, and Os refers to rice.

Fig. 4 shows binary vector, and it is used for the Arabidopis thaliana RBP1 (confidential reference items CDS0078) under rice expression β expansion protein promoter (confidential reference items PRO0061) control.

Fig. 5 shows by β expansion protein promoter driven GUS expression photo." C plant " photo is the GUS dyeing photo when about 5cm size is grown to by rice plant." B plant " photo is the GUS dyeing photo when about 10cm size is grown to by rice plant.Also can use promotor to be used for method of the present invention with similar express spectra.

Fig. 6 has described the example of implementing useful sequence in the method according to this invention in detail.Begin from SEQ ID NO:14, refer to for the At that provides number the MIP accession number ( Http:// mips.gsf.de/); Other identifier refers to the gene pool accession number.Capitalization is represented encoding sequence, and lowercase refers to non-translational region (comprising 5 ' leader sequence, 3 ' non-translational region and intron).The chromosome position of gene is with the coordinate representation of ORF in contig number and the contig.

Embodiment

Describe the present invention with reference to the following example, described embodiment only is intended to explanation.

DNA operation: unless otherwise indicated, recombinant DNA technology is according to being described in (Sambrook etc. (2001) molecular cloning: laboratory manual, the third edition, cold spring harbor laboratory publishes, CSH, New York) or be described in Ausubel etc. (1994), Current Protocols in Molecular Biology, the first roll of Current Protocols and second volume) standard method carry out.The standard material and the method for plant molecular operation are described in Plant Molecular Biology Labfase (1993) by R.D.D.Croy, are published by BIOS Scientific Publications Ltd (UK) and Blackwell ScientificPublications (UK).

Embodiment 1: gene clone-tobacco rna binding protein encoding gene

The protein-bonded gene of coding RNA identifies from tobacco BY2 cell as expressed sequence tag at first and separates as partial sequence that described CDNA-AFLP experiment is used from the cDNA of synchronized tobacco BY2 cell culture (Nicotiana tabacum L.cv.BrightYellow-2) preparation and carried out in the CDNA-AFLP experiment.Based on this CDNA-AFLP experiment, the BY2 label of identification of cell periodic adjustment also selects to be used for further clone.Use the sequence label screening tobacco cDNA library of expression and separate complete sequence cDNA.

The synchronization of BY2 cell

Following by using the aphid rhzomorph of dwelling that cell is arrested in S phase synchronization tobacco BY2 (Nicotiana tabacum L.cv.Bright Yellow-2) culturing cell suspension early.Keep the culturing cell suspension (Int.Rev.Cytol.132 such as Nagata, 1-30,1992) of tobacco Bright Yellow-2 as described.Use 10 times of dilutions of fresh culture to be used for synchronization the static culture in 7 day age, described fresh culture has added aphid dwell rhzomorph (Sigma-Aldrich, St.Louis, MO; 5mg/l), a kind of archaeal dna polymerase α suppresses medicine.After 24 hours, make cell releasing retardance for several times and reenter cell cycle progression with the fresh culture washed cell.

RNA extracts and cDNA synthesizes

(Qiagen, Hilden Germany) extract poly (A according to manufacturer's explanation from total RNA of 500 μ g to use LiCl precipitation preparation total RNA (Sambrook etc., 2001) and use Oligotex post ⁺) RNA.Use biotinylated oligo-dT ₂₅(France) (Life Technologies, Gaithersburg is MD) by the poly (A of reverse transcription from 1 μ g with Superscript II for Genset, Paris for primer ⁺) RNA begins the synthetic first chain cDNA.(USB, Cleveland is OH) with RNA enzyme H (USB) synthesizing by chain replacement carrying out second chain to use intestinal bacteria (Escherichia coli) ligase enzymes (Life Technologies), dna polymerase i.

CDNA-AFLP analyzes

The double-stranded cDNA of 500ng carries out aflp analysis (Vos etc., Nucleic Acids Res.23 (21) 4407-4414,1995 as described; Bachem etc., Plant are (5) 745-53 J.9, and 1996) and modification is arranged.The restriction enzyme of using is BstYI and MseI (Biolabs) and digests in two other steps of branch.Using after one of described enzyme carries out the restrictive diges-tion first time, 3 ' end fragment by its biotinylated tail be collected in the Dyna pearl (Dynal, Oslo, Norway) on, and other fragment is by flush away.Use after second enzymic digestion, collect the restricted fragment of release and it is used as template in follow-up AFLP step.There are not the MseI primer of selective kernel thuja acid and the BstYI combination of primers that contains T or C at 3 ' least significant end to be used for pre-amplification.The PCR condition is (Vos etc., 1995) as described.The amplification mixture that obtains is diluted 600 times, get 5 μ l and be used to use P ³³The BstYI primer of-mark and Amplitaq-Gold polymerase (Roche Diagnostics, Brussels, selective amplification Belgium).In 5% polyacrylamide gel, use the system (Biorad) of Sequigel to separate amplified production.The exsiccant gel on Kodak Biomax film exposure and at phosphoImager (Amersham Pharmacia Biotech, Little Chalfont UK) scans.

The segmental sign of AFLP

To from gel, separate corresponding to the band of differential expression transcript (wherein containing transcript), and under the condition identical, the DNA of wash-out is used for increasing again with selective amplification corresponding to SEQ ID NO 1.By to use polymerase chain reaction product that selectivity BstYI primer increases again directly order-checking or pGEM-T easy (Promega, Madison, WI) in directly order-checking after the cloned sequence, or by the individual clone's acquisition of order-checking sequence information.By Nucleotide and the protein sequence that exists in BLAST sequence alignment (Altschul etc., Nucleic Acids Res.25 (17) 3389-3402 1997) sequence that relatively obtains and the database that can openly obtain.When feasible, the probability that uses long EST or isolating cDNA sequence replacing sequence label to find remarkable homology with increase.Then clone from the physics cDNA of commercial tobacco cDNA amplified library corresponding to SEQ ID NO 1 according to following steps.

Gene clone

Use poly (A ⁺) the average inset of separation is the cDNA library of 1400bp from enliven the not synchronized BY2 tobacco cell of splitted.These library insets are cloned among the carrier pCMVSPORT6.0 (Life Technologies) that contains attB gateway box.From then on select 46000 clones to list in the 384 hole microtiter plates in the library, and follow duplicate place on NF.By using the radiolabeled tag library of the hundreds of kind clone that screening is arranged as probe (wherein containing the BY2 label corresponding to SEQ ID NO1).Separate positive colony (wherein contain with corresponding to the BY2 tag reactant of SEQ IDNO 1 clone), compare to its order-checking and with sequence label.If use label hybridization failure, by the full-length cDNA of following pcr amplification selection corresponding to label.Use primer3 program (http://www-genome.wi.mit.edu/genome_software/other/primer3.html) tag design Auele Specific Primer and be used to the cDNA inset partly that increases with the universal support combination of primers.In pcr amplification, use the dna library of cloning as template from 50000,100000,150000 and 300000 cDNA.Separate amplified production from sepharose, compare to its clone, order-checking and with label.

Then, will be cloned in the suitable plant expression vector via LR Gateway reaction from the full-length cDNA corresponding to SEQ ID NO 1 of pCMVsport6.0 carrier library.

LR gateway reaction is cloned in CDS0701 in the plant expression vector

Then pCMV Sport 6.0p2461 is used for specifying the LR reaction of carrier with the Gateway that is suitable for the rice conversion.This carrier contains within the T-DNA border as the plant selectable marker of functional element and Gateway box, and described Gateway box is used for and has cloned in the aim sequence of donor carrier and carried out recombinating in the LR body.The upstream of this Gateway box is a paddy prolamine promotor, and it is used for the seed-specific expression of gene.

After the reconstitution steps, the expression vector (see figure 2) that produces transformed to enter agrobacterium strains LBA4404 and then transform enter rice plant.

Embodiment 2: rice transforms

Ripe dry seeds shelling with Japanese cultivated rice Nipponbare (NB).By in 70% ethanol, hatching one minute, then at 0.2%HgCl ₂In 30 minutes, the usefulness that continues distillation washing 6 times carried out disinfection in each 15 minutes.Then aseptic seed is being contained 2, sprouting on the substratum of 4-D (callus inducing medium).After around hatching in the dark, downcut embryogenetic scultellum deutero-callus and in identical substratum, breed.After two weeks, breed callus in other words by the other 2 all propagation of succeeding transfer culture in same medium.Before cultivating altogether 3 days, upload to be commissioned to train at fresh culture and support embryo's generation callus fragment (in order to promote the cell fission activity).The agrobacterium strains LBA4404 that will contain double base T-DNA carrier is used for common cultivation.Agrobacterium is inoculated in to contain in the suitable antibiotic AB substratum and at 28 ℃ and cultivated 3 days.Then collect bacterium and be suspended in liquid altogether in the culture medium to about 1 optical density(OD) (OD600).Then suspension is transferred to culture dish and callus was dipped in suspension 15 minutes.Subsequently callus is stained with dry doubling and is transferred to altogether substratum and hatching 3 days in 25 ℃ in the dark of solid on filter paper.In the presence of the selection material of suitable concn, the callus of cultivating is containing 2 altogether, in the substratum of 4-D around 28 ℃ of dark growths.During this period, grow the resistant calli island (resistant callus islands) of quick growth.This substance transfer is hatched under illumination to regeneration culture medium, demonstrate embryo's generation potentiality and grow and sprout in ensuing four to five weeks.Bud is downcut and hatched for 2 to 3 weeks at the substratum that contains growth hormone from callus, with its from this media transfer to soil.The bud of hardening is grown in the greenhouse under the condition in high humidity and short daytime.Then after five months, gather in the crops seed in transplanting three.This method is to provide transformant (Aldemita and Hodges, Planta, 199612-617,1996 of individual gene seat above 50 ratio; Chan etc., Plant Mol.Biol.22 (3) 491-506,1993, Hiei etc., PlantJ., 6 (2) 271-282,1994).

Embodiment 3: assessment and result

15 to 20 independently T0 rice transformants have approximately been produced.In former generation,, transformant transferred to greenhouse growth and results T1 seed by tissue culture room.5 incidents are kept, and wherein separation in 3: 1 of transgenosis existence/shortage take place the T1 offspring.By the expression of monitoring visable indicia, in each incident, select about 10 and contain the T1 seedling of transgenosis (heterozygote and homozygote) and about 10 T1 seedling that lack transgenosis (invalid zygote) of similar number.4 T1 incidents were further assessed according to the appraisal procedure identical with T1 generation in T2 generation, but each incident that described quilt is further assessed has more individuality.

Statistical study: F-check

Use the statistical model of double factor ANOVA (analysis of variance) as plant phenotype characteristic total evaluation.In all plants, all measuring parameters are carried out the F check by all incidents of gene transformation of the present invention.Carry out F and check the validity of checking gene in all transformation events, and checking also is called whole genetic effect to the group effect of gene.With the threshold setting of real whole genetic effect significance is 5% probability level of F check.Significance F test value refers to genetic effect, and it means the difference that the existence of being not only gene or position cause phenotype.

3.1 the measurement of seed correlation parameter

Sophisticated former generation panicle gathered in the crops, packing, slug font code, in 37 ℃ of baking ovens dry three days then.Beat panicle then and to all seed collections and counting.With blowing device full shell is separated with ghost.Abandon ghost, and count rest parts once more.Full shell is weighed on analytical balance.Remaining full hull number is defined as the full seed number after separating step.Measure total seed productive rate by weighing from the whole full shell of plant results.Harvest index among the present invention is defined as total seed productive rate and ground area (mm ²) ratio multiply by the factor 10 ⁶

Following table as a result shows the p value to the F check of T1 and T2 assessment.The difference that also shows percentage ratio between transgenosis and the corresponding invalid zygote.For example, for the total seed weight in T1 generation, 3 in 4 strains are total seed weight positive (promptly compare with the seed weight of corresponding invalid zygophyte, being shown as total seed weight increases (greater than 32%)).2 strains in 4 strains like this show that total seed weight significantly increases, and the p value in the F check is 0.061.

Show the 5:T1 result in generation

T1	Show the product coefficient that increases	Difference	Show the product coefficient that significantly increases	The p value of F check
					Total seed weight	In 43	＞32％	In 42	＜0.061
Harvest index	In 42	＞32％	In 42	＜0.09

Show the 6:T2 result in generation

T2	Show the product coefficient that increases	Difference	Show the product coefficient that significantly increases	The p value of F check
					Total seed weight	In 41	＞30％	In 41	＜0.064

Harvest index

In 41

＞40％

In 41

＜0.001

The gene clone of embodiment 4:AtRBP1

(Invitrogen, Paisley UK) pass through pcr amplification Arabidopis thaliana AtRBP1 (CDS0078) as template to use Arabidopis thaliana seedling cDNA library.After the RNA reverse transcription that seedling is extracted, cDNA is cloned into pCMV Sport 6.0.The average inset size in library is 1.5kb, and original clone's number is 1.59 * 10 ⁷Cfu.Original titre is confirmed as 9.6 * 10 ⁵Cfu/ml is 6 * 10 after amplification for the first time ¹¹Cfu/ml.Plasmid is used for 50 μ l PCR mixtures with the 200ng template after extracting.The primer prm00405 (justice 5 ' ggggacaagtttgtacaaaaaagcaggcttcacaatggattatgatcggtacaagt tat 3 ') and the prm00406 (reverse complemental: 5 ' ggggaccactttgtacaagaaagctgggtttaaaagagtccaaagaatttcact 3 ') be used to pcr amplification that comprise the AttB site that is used for the Gateway reorganization.Under standard conditions, use Hifi Taq archaeal dna polymerase to carry out PCR.The PCR fragment of 1209bp is amplified and also uses the standard method purifying.Then carry out the first step of Gateway operation, the BP reaction, generation " entering the clone " p0073 according to the Gateway term during this period recombinates in PCR fragment and the pDONR201 plasmid body.Plasmid pDONR201 as the part of Gateway  technology available from Invitrogen.

Embodiment 5:AtRBP1 vector construction

Then will enter clone p0073 is used for reacting with the LR of p03069 (being used for the appointment carrier that rice transforms).This carrier contains as functional element within the T-DNA border: plant selectable marker; Visable indicia expression cassette and Gateway box, described Gateway box are used for and have cloned in the aim sequence of " entering the clone " and carried out reorganization in the LR body.The β expansion protein promoter of expressing in bud is positioned at the upstream of this Gateway box.

After the LR reconstitution steps, the expression vector (Fig. 2) that produces transformed to enter agrobacterium strains LBA4404 and then transform enter rice plant.Make rice plant's growth of conversion and follow inspection in the parameter described in the embodiment 6.

The assessment of embodiment 6:AtRBP1 and result

15 to 20 independently T0 rice transformants have approximately been produced.In former generation,, transformant transferred to greenhouse growth and results T1 seed by tissue culture room.5 incidents are kept, and wherein separation in 3: 1 of transgenosis existence/shortage take place the T1 offspring.By the expression of monitoring visable indicia, in each incident, select about 10 and contain the T1 seedling of transgenosis (heterozygote and homozygote) and about 10 T1 seedling that lack transgenosis (invalid zygote) of similar number.4 T1 incidents were further assessed according to the appraisal procedure identical with T1 generation in T2 generation, but each incident that described quilt is further assessed has more individuality.In the first round is that strain of neutral no longer continues.In T2 assessment, what relatively contain genetically modified 15 T2 seedling and similar number lacks genetically modified plant (invalid zygote).

Statistical study: F-check

Use the statistical model of double factor ANOVA (analysis of variance) as plant phenotype characteristic total evaluation.In all plants, all measuring parameters are carried out the F check by all incidents of gene transformation of the present invention.Carry out F and check the validity of checking gene in all transformation events, and the whole structure of checking gene, whole genetic effect also be called.With the threshold setting of real whole genetic effect significance is 5% probability level of F check.Significance F test value refers to genetic effect, and it means the difference that the existence of being not only gene or position cause phenotype.

6.1 the measurement of seed correlation parameter

Sophisticated former generation panicle gathered in the crops, packing, slug font code, in 37 ℃ of baking ovens dry three days then.Beat panicle then and to all seed collections and counting.With blowing device full shell is separated with ghost.Abandon ghost, and count rest parts once more.Full shell is weighed on analytical balance.This operation produces seed correlation parameter group described below.

The following demonstration of table as a result is from the p value of the F check that T1 assessment, T2 are assessed and the combination p value of checking from the F to T1 and T2 assessment.Combinatory analysis can be thought same incident is carried out two experiments.The confidence level that this can be used to detect the consistence of two experiment effects and increase conclusion.The method of using is the mixed mode method, and it considers multilevel data structure (i.e. experiment-incident-segregant).Probability Detection by relatively card side's distribution obtains the P value.Each table give per generation transgenosis and corresponding invalid zygote between % difference.

6.1.1 ground area

Sum of all pixels by ground plant part after the calculating eliminating background is determined plant area on the ground.This value is to put the mean value that obtains picture from different perspectives at one time, and is converted into the physical surface value of representing with square millimeter by calibration.Experiment shows that the over-ground part plant area of this method measurement is relevant with the biomass that plant shoot divides.The results are shown in the following table 7 of T1 and T2 assessment.As shown in the table, be significance from the p value (the p value is 0.0011) of the F check that T2 is assessed and the data (having the p value is 0.0287) of combination, the construct that its explanation exists in plant has remarkable positive the influence to the ground area of transgenic plant.

Table 7: ground area

The ground area
				% difference	The P value
T1 is overall	8	0.1779
			T2 is overall	15	0.0011
Combination		0.0012

6.1.2 the total seed production of every strain plant

Measure total seed production by weighing from the whole full shell of plant results.As shown in table 8 below, the p value of checking from the F to T1 and T2 combined evaluation is significance (the p value is 0.0287), and the existence of its explanation construct in plant has the significance influence to total seed weight of transgenic plant.

Table 8

Total seed weight
				% difference	The P value

T1	12	0.3397
			T2	16	0.1356
Combination		0.0287

6.1.3 seed sum

As shown in table 9 below, the p value of checking from the F to T1 and T2 combination (and T2 is independent) assessment is significance (the p value is 0.0006), and the existence of its explanation construct in plant has the significance influence to the seed sum of transgenic plant.

Table 9

The seed sum
				% difference	The P value
T1	6	0.4044
			T2	23	0.0003
Combination		0.0006

Embodiment 7: expressed by β expansion protein promoter driven GUS

Using " BP recombining reaction " that β is expanded protein promoter clones in the pDONR201 of GatewayTM system and enters plasmid (Life Technologies).The identity and the base pair of the inset of determining the clone by checking order are formed, and additionally detect the plasmid that produces via restrictive diges-tion.

For promotor being cloned in before the reporter gene, then each being entered the clone and be used for " LR recombining reaction " (GatewayTM) with specifying carrier.Design effectively is connected in intestinal bacteria β glucuronidase (GUS) gene with this appointment carrier, and described connection is carried out via the replacement of the reorganization of the Gateway before gus gene box.Then use the standard conversion technology that the report carrier (comprising the promotor that effectively is connected in GUS) that produces is transformed the conversion that enters agrobacterium strains LBA4044 and continue and enter rice plant.

Produce transgenosis rice plant from cell transformed.Plant-growth is carried out under normal operation.

Use 90% ice-cold acetone to cover plant or plant part to be tested, hatched 30 minutes at 4 ℃.[15.76g Trizma HCl (Sigma T3253)+2.922g NaCl is in 1 liter of distilled water to use the Tris damping fluid, use NaOH to be adjusted to pH7.0] wash 3 times, after each 5 minutes, this material is covered with Tris/ ferricyanate/X-Gluc solution [9.8ml Tris damping fluid+0.2ml ferricyanate mother liquor (0.33g potassium ferricyanate (Sigma P3667) is in 10ml Tris damping fluid)+0.2ml X-Gluc mother liquor (26.1mg X-Gluc (Europa Bioproducts ML 113A) is in 500 μ l DMSO)].Vacuum filtration 15 to 30 minutes.Hatching plant or plant part at 37 ℃ reaches 16 hours and develops as seen until blueness.With Tris damping fluid washing sample 3 times 5 minutes.Ethanol series with 50%, 70% and 90% is extracted chlorophyll (each is 30 minutes).

Sequence table

＜110〉Cropdesign NV

＜120〉has the plant and preparation method thereof of improvement growth characteristics

<130>CD-120-PCT

<150>EP 04103926.4

<151>2004-08-16

<150>US 60/602,680

<151>2004-08-19

<160>40

<170>PatentIn version 3.3

<210>1

<211>2098

<212>DNA

＜213〉tobacco (Nicotiana tabacum)

<400>1

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agacttccat tgttcccagc ggctaagatg agccggttga ttgagcatca cctagcaaat 120

aataaacagg acatgaaagg gacagaggtt tttgttggtg gtttggcccg tactactact 180

gaaagcaaaa ttcatgaggt attttcttca tgtggtgaga ttgtggaaat acggttgata 240

aaagaccaga caggcgttcc taaggggttt tgctttgtac gatttgcaac aaaatatgct 300

gctgacaaag ctctgaagga aaaatctgga tatgtgctgg atgggaagaa actcggggtt 360

cgcccctcag ttgagcagga cactttattt cttggaaatc ttaacaaagg ttggggtgcg 420

gaggaatttg agagtattgt gcgccaggtt tttccagatg ttgtatctgt tgatcttgca 480

cttcttggag atgtccaacc tggtcagaag caacggaatc ggggttttgc tttcgtgaaa 540

ttcccatctc atgctgctgc ggctcgtgct tttcgggtag gctcccaatc tgattttctc 600

attgatggca agttacatcc atctgtacag tgggctgagg aacctgatcc caatgaactt 660

gctcagatca aagcagcctt cgttagaaat gtacctcctg gtgctgatga agattacttg 720

aagaagctct ttcagccctt tggcaatgta gagaggatag ctctatccag gaaaggtagc 780

tccaccattg gattcgttta cttcgataag cgatctgatc ttgacaatgc tattatggcg 840

ttgaatgaga aaactgtaca agggccaatg ggaggtccct catgcaagct tcaggtcgaa 900

gttgctaggc caatggacaa gaacaggaaa cgaggtcgtg aggatccaaa catgtccagt 960

accattgaga gtcattccaa gcttttgaag gatgatccag atgttgagat gattagggct 1020

cctaaatcaa ctgctcaact ggagatggat tattcggatc cttatgaagc tgctgtagtt 1080

gcattacctg tggttgtcaa ggagcgttta gttcggatct tgcggcttgg tattgctact 1140

agatatgata tagatgttga aagtttaacc agtcttaaga tattgcccca gtcagctgcc 1200

atatctattc ttgaccagtt catgttgtct ggagctgata tgcagaacaa gggaggatat 1260

ctagcttcat taatttctaa gcaggttgaa aaactgggac cgaaacaatt cgatagtagg 1320

tcaaggatag aagatgttgg cttgagggtg ccagaaccag acaggttctc tacaagagtt 1380

cgtttgccag atctagattc atatgcctca cgagtaccct tgcccatgcc taggactgat 1440

gtttacacat ctcactattc agcgtattta gatccccatc tgtctggtcg gatgacagca 1500

aagaggatgg aggaagcaag ttcccatttg caggcgactt cacttctgtc tagtcgggtg 1560

gcaacgagga tggaggaggc aggttccact ttgcagtcgc tcctatctgg tggggtgacg 1620

acaagaagga tggaggaagc aagtccgatt ttgcaggcaa cactccttcc atctggtcgg 1680

gtatcaagga tggatgaagc aagtcccaat ttgcaggcaa catggagccc ttctcctact 1740

aatgacagaa ttggacttca ttcacacatt accgcaactg ctgatcatca acatactcga 1800

ccacggatca ggtttgatcc cttcactggt gagccataca aatttgaccc cttcactggc 1860

gagccaattg ttcccaagag ctcaagtcat catcgaagcc tgtactgaac gttctgagca 1920

ttctaattta caaatggctt attgccaaac ctatgtaaca taatgatgcg tatttttgtt 1980

catccgcagc tgtaaaatag tagctgttag caggattatt tggttatgtt tctcattgac 2040

ttcattgatt gcgaaggtgc atttggaatc tcggcaatca caatttatag ccggtgca 2098

<210>2

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＜213〉tobacco

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Gly Tyr Val Leu Asp Gly Lys Lys Leu Gly Val Arg Pro Ser Val Glu

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115 120 125

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130 135 140

Arg Gly Phe Ala Phe Val Lys Phe Pro Ser His Ala Ala Ala Ala Arg

145 150 155 160

Ala Phe Arg Val Gly Ser Gln Ser Asp Phe Leu Ile Asp Gly Lys Leu

165 170 175

His Pro Ser Val Gln Trp Ala Glu Glu Pro Asp Pro Asn Glu Leu Ala

180 185 190

Gln Ile Lys Ala Ala Phe Val Arg Asn Val Pro Pro Gly Ala Asp Glu

195 200 205

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210 215 220

Ala Leu Ser Arg Lys Gly Ser Ser Thr Ile Gly Phe Val Tyr Phe Asp

225 230 235 240

Lys Arg Ser Asp Leu Asp Asn Ala Ile Met Ala Leu Asn Glu Lys Thr

245 250 255

Val Gln Gly Pro Met Gly Gly Pro Ser Cys Lys Leu Gln Val Glu Val

260 265 270

Ala Arg Pro Met Asp Lys Asn Arg Lys Arg Gly Arg Glu Asp Pro Asn

275 280 285

Met Ser Ser Thr Ile Glu Ser His Ser Lys Leu Leu Lys Asp Asp Pro

290 295 300

Asp Val Glu Met Ile Arg Ala Pro Lys Ser Thr Ala Gln Leu Glu Met

305 310 315 320

Asp Tyr Ser Asp Pro Tyr Glu Ala Ala Val Val Ala Leu Pro Val Val

325 330 335

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340 345 350

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355 360 365

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Pro Ile Val Pro Lys Ser Ser Ser His His Arg Ser Leu Tyr

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<210>3

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<212>DNA

＜213〉rice (Oryza sativa)

<400>3

atggtgcgtg ctcgagactc aatccgcgaa atcctccctg ttttttcgat tcaatccgcc 60

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cgtgggggat ttcaggagaa gaggggggcg gcgtcgcatg gcgactacga cgagcaaggt 240

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gcgccgaggt gttcgtcggc gggttgccgc ggtcggtgac ggagcgggcg ctccgagagg 360

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aagtttgatc cctacaccgg tgaacccatc aggccagaat cgaacccacg tcgctcagga 1020

agcttatact gactttgatt gattgaagca acagtttgga tatggtagat tagatttaca 1080

tccctgaacc aaaaggacca tat 1103

<210>4

<211>680

<212>PRT

＜213〉rice

<400>4

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20 25 30

Val Ala Ala Ala Ser Asp Leu Val Arg Ile Ser Ser Glu Lys Ser Arg

35 40 45

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Gly Asn Leu Cys Lys Gly Ser Asp Trp Gly Ile Glu Glu Phe Glu Glu

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Leu Ile Arg Lys Val Arg Pro Val Val Asp Leu Ala Met Ala Arg Asn

195 200 205

His Asp Ser Ser Val Gly Lys Arg Arg Leu Asn Arg Gly Phe Ala Phe

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Val Arg Phe Ser Ser His Ala Val Ser Gln Val Lys Thr Ala Phe Val

225 230 235 240

Gly Asn Leu Pro Ala Asn Val Thr Glu Glu Tyr Leu Arg Lys Leu Phe

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Glu His Cys Gly Glu Val Cys Tyr Ala Val Val Arg Val Ala Val Ser

260 265 270

Arg Lys Gly Gln Tyr Pro Val Gly Phe Val His Phe Ala Ser Arg Thr

275 280 285

Trp Lys Glu Leu Asp Asn Ala Ile Lys Glu Met Asp Gly Glu Thr Val

290 295 300

Arg Gly Pro Asp Arg Gly Ala Thr Phe Arg Ile Gln Val Ser Val Ala

305 310 315 320

Arg Pro Val Val Glu Asn Asp Lys Lys Arg Ile Arg Glu Glu Val Lys

325 330 335

Thr Arg Arg Ser Asn Val Ser Thr Asp Lys Pro Asp His Ser Tyr Gly

340 345 350

Arg Arg Gly His Asp Ser Tyr Asp Arg Gln Ala Lys Ala Pro Arg Leu

355 360 365

Tyr Asn Glu Val Leu His Thr Asn Asp Lys Val Asp Met Ile Thr Leu

370 375 380

Phe Cys Gln Phe Ser Phe Leu Gln Val Ser Asp Thr Asp Pro Tyr Glu

385 390 395 400

Ala Ala Val Val Ser Leu Pro Ser Ala Val Lys Glu Leu Leu Leu Arg

405 410 415

Ile Leu Arg Leu Arg Ile Gly Thr Arg Tyr Asp Val Ser Asn Leu Tyr

420 425 430

Ile Arg Ser Leu Leu Val Ser Ile Leu Leu Phe Gln Ile Asp Ile His

435 440 445

Cys Ile Arg Ser Leu Asn Glu Leu Pro Glu Lys Ala Ala Val Ala Val

450 455 460

Leu Asn Gln Cys Ser Gln Phe Leu Ile Ser Gly Ala Asp Lys His Asn

465 470 475 480

Lys Gly Asp Tyr Phe Ala Ser Leu Ile Ala Lys Glu Thr Phe Ser Ser

485 490 495

Ala Leu Arg Leu Gln Gly Ser Thr Tyr Leu Pro Arg Asn Pro Glu Ile

500 505 510

Gln Asn Lys Arg Phe Pro His Ser Ser Arg Tyr Ser Ser Leu Gly Asp

515 520 525

Tyr Pro Ser Ser Ser Tyr Val Asp Asp Pro Ala Ser Ser Gln Ser Arg

530 535 540

Asn Arg Arg Tyr Asp Glu Tyr Arg Pro Asp Leu Val Arg Tyr Pro Asp

545 550 555 560

Ser Arg Ser Arg Gln Glu Glu Ile Val Arg Ile Glu Arg Tyr Pro Glu

565 570 575

Pro Arg Phe Ala His Glu Pro Arg Gln Asp Thr Gly Arg His Leu Asp

580 585 590

Leu Gly Tyr Val Gln Glu Arg Asn Ser Asn Ile Glu Arg Ser Ala Gln

595 600 605

Val Ala Phe Ser Ser Arg Glu Gly Gly Tyr Leu Ser Ala Ser Arg Tyr

610 615 620

Asn Thr Asn Ile Val Pro Glu Phe Ser Ser Arg Ser Ser Ala Glu Tyr

625 630 635 640

Ser Thr Ala Arg Gln Gln Val Arg Phe Asp Pro Phe Thr Gly Glu Pro

645 650 655

Tyr Lys Phe Asp Pro Tyr Thr Gly Glu Pro Ile Arg Pro Glu Ser Asn

660 665 670

Pro Arg Arg Ser Gly Ser Leu Tyr

675 680

<210>5

<211>1758

<212>DNA

＜213〉corn (Zea mays)

<220>

<221>misc_feature

<222>(1632)..(1632)

＜223〉n is a, c, g or t

<220>

<221>misc_feature

<222>(1699)..(1699)

＜223〉n is a, c, g or t

<220>

<221>misc_feature

<222>(1722)..(1722)

＜223〉n is a, c, g or t

<220>

<221>misc_feature

<222>(1747)..(1747)

＜223〉n is a, c, g or t

<220>

<221>misc_feature

<222>(1754)..(1754)

＜223〉n is a, c, g or t

<400>5

tctagctgtg ttcttgtggc tgtgaaatta tatctcccat gctgatactt gattccctta 60

tctttgcttc attactacac cacagtaatt tggatctgcc attatgttac tatgtaactc 120

tcatttgata tcaatcacag ctgccacata caaaatacaa gtatgtttat ctagataaga 180

tcttgattca tcaatcacca ctgatctgag ttttcgccac tgcgatgcga ggaaaagaca 240

gatatctaat aacatcttgg tgaagatgtt cttaggtcct ttgctttctc ttcaagtcag 300

cttcctttga tttcattcct caaactatca atcacaggct gcagcacgtg taatccgcat 360

cggttcaaga acagatttca tgcttggtga tattttgcat cctgcgataa attgggctga 420

taaagagtct catctggatc ctgatgaaat ggccaagatg aagtctgctt ttattggtaa 480

cctgccagaa gatgttaatg aggagtactt gagaaagctt tttggacagt tcggtgaggt 540

agtacgggtt gctatctcaa gaaaaggaca atgtccagtt gcttttgttc acttcgccaa 600

acgttcagag cttgagaatg ctatagaaga aatggatggt aaaacggtga gaggacctgg 660

tcgagggccg tctttcaaga tccaggtgtc agttgctcga cctacggcag acaacgacaa 720

gaagcgatct cgtgaagaag tgagaactag aagatcaaat gcatcaggag ataggcgaga 780

ttattctcat ggaagatatg gacacgattc acttgatcgt caagtgaaag ctccaagatt 840

atctaattat gtggccgatg ctgctgaccc ctatgaatca gctgttaatt cattaccttc 900

agctgtcaag gaagtcttgc ttcgaattct acgtctaaga attggtactc gatatgatat 960

tgatatccat tgtgttaaaa gccttgatga gcttcctgag tcatctgctc ttgctgtcct 1020

taatcagttt ttgatatcag gtggagacaa acacaacaaa ggagattatt ttgcatcgtt 1080

ggttgctaag caccaggctg agacctttgg cttaacacat gcattacacg gtaccactta 1140

tttgtcaaga aatccggaaa tgcatagcaa gcgataccca catgaagatt atgattttgt 1200

gacacccagg agcagtaggt acgattcgtc agcccatcat ccttcaacat actacgaaga 1260

cgatccacca gtgtctgagt caagggttag aagatatgct gaagaaaggt ccaccattgt 1320

aagaagccca gaaccacgtc cgcgatatga cgaaacagac ataagaataa acccagaacc 1380

aagattacca tatgaatcaa gacacaacgc cgaaaagcat ctcgatcgaa gatacataca 1440

agagcatagt tcaaatattg aaagaccagc tgaagaagct ctcctttcta gggaaaggag 1500

atttctgcct gctgcagggt acatgccgaa cccaggcggc tcggatttcc gctccaggtc 1560

gcccgccgaa tattcagcac aacgccaaca aatgaggttt gatccattca caggtgaacc 1620

ttacaagttt gnacccttca caggggagcc catcaggcca gatccgaacc cagcgccgct 1680

caggaagcct gtaattgant cagaataagt ttggaagccg anaatgccag attaagaacc 1740

ctgaaancaa agcnaaga 1758

<210>6

<211>448

<212>PRT

＜213〉corn

<220>

<221>misc_feature

<222>(424)..(424)

＜223〉Xaa can be any naturally occurring amino acid

<220>

<221>misc_feature

<222>(446)..(446)

＜223〉Xaa can be any naturally occurring amino acid

<400>6

Gly Ser Arg Thr Asp Phe Met Leu Gly Asp Ile Leu His Pro Ala Ile

1 5 10 15

Asn Trp Ala Asp Lys Glu Ser His Leu Asp Pro Asp Glu Met Ala Lys

20 25 30

Met Lys Ser Ala Phe Ile Gly Asn Leu Pro Glu Asp Val Asn Glu Glu

35 40 45

Tyr Leu Arg Lys Leu Phe Gly Gln Phe Gly Glu Val Val Arg Val Ala

50 55 60

Ile Ser Arg Lys Gly Gln Cys Pro Val Ala Phe Val His Phe Ala Lys

65 70 75 80

Arg Ser Glu Leu Glu Asn Ala Ile Glu Glu Met Asp Gly Lys Thr Val

85 90 95

Arg Gly Pro Gly Arg Gly Pro Ser Phe Lys Ile Gln Val Ser Val Ala

100 105 110

Arg Pro Thr Ala Asp Asn Asp Lys Lys Arg Ser Arg Glu Glu Val Arg

115 120 125

Thr Arg Arg Ser Asn Ala Ser Gly Asp Arg Arg Asp Tyr Ser His Gly

130 135 140

Arg Tyr Gly His Asp Ser Leu Asp Arg Gln Val Lys Ala Pro Arg Leu

145 150 155 160

Ser Asn Tyr Val Ala Asp Ala Ala Asp Pro Tyr Glu Ser Ala Val Asn

165 170 175

Ser Leu Pro Ser Ala Val Lys Glu Val Leu Leu Arg Ile Leu Arg Leu

180 185 190

Arg Ile Gly Thr Arg Tyr Asp Ile Asp Ile His Cys Val Lys Ser Leu

195 200 205

Asp Glu Leu Pro Glu Ser Ser Ala Leu Ala Val Leu Asn Gln Phe Leu

210 215 220

Ile Ser Gly Gly Asp Lys His Asn Lys Gly Asp Tyr Phe Ala Ser Leu

225 230 235 240

Val Ala Lys His Gln Ala Glu Thr Phe Gly Leu Thr His Ala Leu His

245 250 255

Gly Thr Thr Tyr Leu Ser Arg Asn Pro Glu Met His Ser Lys Arg Tyr

260 265 270

Pro His Glu Asp Tyr Asp Phe Val Thr Pro Arg Ser Ser Arg Tyr Asp

275 280 285

Ser Ser Ala His His Pro Ser Thr Tyr Tyr Glu Asp Asp Pro Pro Val

290 295 300

Ser Glu Ser Arg Val Arg Arg Tyr Ala Glu Glu Arg Ser Thr Ile Val

305 310 315 320

Arg Ser Pro Glu Pro Arg Pro Arg Tyr Asp Glu Thr Asp Ile Arg Ile

325 330 335

Asn Pro Glu Pro Arg Leu Pro Tyr Glu Ser Arg His Asn Ala Glu Lys

340 345 350

His Leu Asp Arg Arg Tyr Ile Gln Glu His Ser Ser Asn Ile Glu Arg

355 360 365

Pro Ala Glu Glu Ala Leu Leu Ser Arg Glu Arg Arg Phe Leu Pro Ala

370 375 380

Ala Gly Tyr Met Pro Asn Pro Gly Gly Ser Asp Phe Arg Ser Arg Ser

385 390 395 400

Pro Ala Glu Tyr Ser Ala Gln Arg Gln Gln Met Arg Phe Asp Pro Phe

405 410 415

Thr Gly Glu Pro Tyr Lys Phe Xaa Pro Phe Thr Gly Glu Pro Ile Arg

420 425 430

Pro Asp Pro Asn Pro Ala Pro Leu Arg Lys Pro Val Ile Xaa Ser Glu

435 440 445

<210>7

<211>1599

<212>DNA

＜213〉rice

<400>7

atcgatcaca ggctgcagca cgcgtacttc gtattggttc cagaacagat tttctgcttg 60

gtggattgca tccttcaata aattgggctg agaaggagtc tcatgtagat gaggacgaaa 120

tggccaaggt taagacagct ttcgttggaa atttaccagc aaatgttaca gaggagtatt 180

taagaaagct ttttgaacat tgtggagagg tagtacgggt tgcagtctca aggaaaggac 240

aatatccagt tggatttgtc cactttgcca gtcgtacaga gctcgacaat gcaataaaag 300

aaatggatgg tgaaacagtg agaggacctg accgaggggc aactttcagg atccaggtct 360

cagttgctcg gcctgtggta gagaacgata aaaagagaat tcgtgaagaa gtgaaaacta 420

gaagatcaaa cgtatcaaca gacaagccgg accattctta tggaagacgt ggacatgatt 480

catatgatcg tcaagcaaaa gctccaaggc tatataatga ggtgtctgat acggacccct 540

atgaagcagc tgttgtttca ctaccttcag ccgtcaagga actcctactt cgtattctac 600

gtcttagaat tggcactcga tatgatatag acattcattg cataaggagt cttaatgaac 660

ttcctgaaaa ggctgcagtt gctgtcctta atcagttttt gatatcaggt gcagataaac 720

acaataaagg agactatttc gcttcattaa ttgctaagta ccaggctgag acatttagct 780

cagcactaag attgcagggt tctacttatt tgccaagaaa tcctggaata cagaacaaga 840

gattcccaca tcaagattac gagtacacag catccgggag tagtagatac agttccttag 900

gtgattatcc ttcctcatct tatgtggatg atcccgcatc atctcagtca aggaatagaa 960

ggtatgatga atacagacct gatcttgtaa gatatccaga ttcaagatca cggcaagagg 1020

aaatagtccg cattgaaaga tatccagaac caagatttgc acatgaacca agacaggata 1080

ctggaaggca tctcgatcta gggtacgtac aagaacggaa ttcgaatatt gagagatcag 1140

ctcaagtagc tttttcatct agggaaggag gatacttatc tgcttcaagg tacaacacaa 1200

acatagtccc agaattcagc tccaggtcat ctgctgaata ctctactgct cgccaacaag 1260

taaggtttga tccattcaca ggggaaccat acaagtttga tccctacacc ggtgaaccca 1320

tcaggccaga atcgaaccca cgtcgctcag gaagcttata ctgactttga ttgattgaag 1380

caacagtttg gatatggtag attagattta catccctgaa ccaaaaggac catatactgc 1440

tcttgcatgt tgtaaaccta gtgtatttga tgtgcctcag cattgtaatg ttagaaatcc 1500

attttcatcc atgtcactgg aaaactatgg ttgaaacaac agtaataagt tctatcattt 1560

atgatggcat ctgatgatat gaattaggga aaactaagc 1599

<210>8

<211>414

<212>PRT

＜213〉rice

<400>8

Met Ala Lys Val Lys Thr Ala Phe Val Gly Asn Leu Pro Ala Asn Val

1 5 10 15

Thr Glu Glu Tyr Leu Arg Lys Leu Phe Glu His Cys Gly Glu Val Val

20 25 30

Arg Val Ala Val Ser Arg Lys Gly Gln Tyr Pro Val Gly Phe Val His

35 40 45

Phe Ala Ser Arg Thr Glu Leu Asp Asn Ala Ile Lys Glu Met Asp Gly

50 55 60

Glu Thr Val Arg Gly Pro Asp Arg Gly Ala Thr Phe Arg Ile Gln Val

65 70 75 80

Ser Val Ala Arg Pro Val Val Glu Asn Asp Lys Lys Arg Ile Arg Glu

85 90 95

Glu Val Lys Thr Arg Arg Ser Asn Val Ser Thr Asp Lys Pro Asp His

100 105 110

Ser Tyr Gly Arg Arg Gly His Asp Ser Tyr Asp Arg Gln Ala Lys Ala

115 120 125

Pro Arg Leu Tyr Asn Glu Val Ser Asp Thr Asp Pro Tyr Glu Ala Ala

130 135 140

Val Val Ser Leu Pro Ser Ala Val Lys Glu Leu Leu Leu Arg Ile Leu

145 150 155 160

Arg Leu Arg Ile Gly Thr Arg Tyr Asp Ile Asp Ile His Cys Ile Arg

165 170 175

Ser Leu Asn Glu Leu Pro Glu Lys Ala Ala Val Ala Val Leu Asn Gln

180 185 190

Phe Leu Ile Ser Gly Ala Asp Lys His Asn Lys Gly Asp Tyr Phe Ala

195 200 205

Ser Leu Ile Ala Lys Tyr Gln Ala Glu Thr Phe Ser Ser Ala Leu Arg

210 215 220

Leu Gln Gly Ser Thr Tyr Leu Pro Arg Asn Pro Gly Ile Gln Asn Lys

225 230 235 240

Arg Phe Pro His Gln Asp Tyr Glu Tyr Thr Ala Ser Gly Ser Ser Arg

245 250 255

Tyr Ser Ser Leu Gly Asp Tyr Pro Ser Ser Ser Tyr Val Asp Asp Pro

260 265 270

Ala Ser Ser Gln Ser Arg Asn Arg Arg Tyr Asp Glu Tyr Arg Pro Asp

275 280 285

Leu Val Arg Tyr Pro Asp Ser Arg Ser Arg Gln Glu Glu Ile Val Arg

290 295 300

Ile Glu Arg Tyr Pro Glu Pro Arg Phe Ala His Glu Pro Arg Gln Asp

305 310 315 320

Thr Gly Arg His Leu Asp Leu Gly Tyr Val Gln Glu Arg Asn Ser Asn

325 330 335

Ile Glu Arg Ser Ala Gln Val Ala Phe Ser Ser Arg Glu Gly Gly Tyr

340 345 350

Leu Ser Ala Ser Arg Tyr Asn Thr Asn Ile Val Pro Glu Phe Ser Ser

355 360 365

Arg Ser Ser Ala Glu Tyr Ser Thr Ala Arg Gln Gln Val Arg Phe Asp

370 375 380

Pro Phe Thr Gly Glu Pro Tyr Lys Phe Asp Pro Tyr Thr Gly Glu Pro

385 390 395 400

Ile Arg Pro Glu Ser Asn Pro Arg Arg Ser Gly Ser Leu Tyr

405 410

<210>9

<211>1842

<212>DNA

＜213〉rice

<400>9

atggaaccga cgcgccgttg cgtccccggc catctcgcca ccgccgccgc cgccgccgcc 60

gcctcgccgt tctccccgcc gccgtcgctg ccgctgccgt ccgcgctcat gccccccaag 120

aagcgccgcc tcttcacgcc cgcccctcgc cacgccgcca ccccgccacc accaccacct 180

ccccccaccc ccgccgtcga gcccacccta ccaatccccc ccgcctcgac accgccgacg 240

ccgcctcagc cctccgcctc cacggagccc tcgacggcgc cgcctcccgc tgtcgacgac 300

gcggcggcga ggtcgtcgtc gtcgtcgtcg ccggcgtcgg cggcggcggc gcggaaggtt 360

cggaaagtgg ttaagaaggt catcgtcaag aaggtcgtcc ccaagggcac gttcgccgct 420

cggaaggccg cggcggcggc ggttgctgct gctgcggcgg tctccggagc agcagcatca 480

tcggaggcag ggggagaagc cccaaccgac gagccagcaa gtgatcagga cggcggagtt 540

gggaatgagc aaaaattgga tgaatccaaa cctgccacgg attgcaatgc cgttgcggtg 600

gtggaagaat cggtgtgtaa ggaggaggag gaggtggcct tagtggtggg taagggagtg 660

gaggaggagg aggcggggat gtcggagcgg cggaagagga tgaccatgga ggtgtttgtt 720

ggtgggcttc accgggacgc caaggaggat gatgtgaggg cggtgttcgc caaggccggg 780

gaaatcaccg aggtccggat gataatgaat cctcttgcag ggaagaacaa ggggtactgc 840

ttcgtgcgct accgccacgc cgcgcaggcg aagaaggcca tcgcggaatt cggcaatgtg 900

aagatttgtg ggaagctctg tcgagctgca gttccagttg ggaatgacag aatttttctt 960

ggaaacatca acaagaaatg gaaaaaagaa gatgtcatca agcagctaaa gaaaattgga 1020

attgagaaca ttgattctgt aacacttaag tctgattcaa ataatccagt ctgtaatcgt 1080

ggttttgcat ttcttgaact ggaaactagt agagatgcac ggatggcata caaaaagctt 1140

tcacagaaaa atgcttttgg caaaggcctg aatataagag ttgcatgggc tgaaccattg 1200

aatgatccag atgagaaaga tatgcaggtt aaatcgattt ttgtggatgg gataccaacg 1260

tcctgggatc atgctcagct aaaagaaatc ttcaagaaac atgggaagat tgaaagtgtg 1320

gttctgtcac gcgatatgcc gtcagctaaa aggagggact ttgcctttat taattacatt 1380

actcgtgagg ctgcaatctc gtgtcttgaa tcttttgaca aggaagagtt cagtaagaac 1440

ggctcaaagg tgaatattaa agtttcattg gctaaacctg cccaacagag caagcagacc 1500

aaggaagacc ataaatctag tattactggg gaaggcaaaa tgaagacttc taaaataaga 1560

taccctgttc aagattatac ccacatttat tctggagaga agcgtccctt ttcaacactg 1620

ggtgatcctt attatccatt gagaggtcat tcttgtcgtc gtcatgaggg tagcacctat 1680

actacagcag catcaagcta tggtgcgctg ccccctgcta ctgctgaatc ttctctgcca 1740

cattatcatg acagcaatag atatcctcca cacctaggtg aggcaatcaa gttctcgcca 1800

accagcgcag tcctatcgaa gcaggcatgg caaaaaatgt aa 1842

<210>10

<211>613

<212>PRT

＜213〉rice

<400>10

Met Glu Pro Thr Arg Arg Cys Val Pro Gly His Leu Ala Thr Ala Ala

1 5 10 15

Ala Ala Ala Ala Ala Ser Pro Phe Ser Pro Pro Pro Ser Leu Pro Leu

20 25 30

Pro Ser Ala Leu Met Pro Pro Lys Lys Arg Arg Leu Phe Thr Pro Ala

35 40 45

Pro Arg His Ala Ala Thr Pro Pro Pro Pro Pro Pro Pro Pro Thr Pro

50 55 60

Ala Val Glu Pro Thr Leu Pro Ile Pro Pro Ala Ser Thr Pro Pro Thr

65 70 75 80

Pro Pro Gln Pro Ser Ala Ser Thr Glu Pro Ser Thr Ala Pro Pro Pro

85 90 95

Ala Val Asp Asp Ala Ala Ala Arg Ser Ser Ser Ser Ser Ser Pro Ala

100 105 110

Ser Ala Ala Ala Ala Arg Lys Val Arg Lys Val Val Lys Lys Val Ile

115 120 125

Val Lys Lys Val Val Pro Lys Gly Thr Phe Ala Ala Arg Lys Ala Ala

130 135 140

Ala Ala Ala Val Ala Ala Ala Ala Ala Val Ser Gly Ala Ala Ala Ser

145 150 155 160

Ser Glu Ala Gly Gly Glu Ala Pro Thr Asp Glu Pro Ala Ser Asp Gln

165 170 175

Asp Gly Gly Val Gly Asn Glu Gln Lys Leu Asp Glu Ser Lys Pro Ala

180 185 190

Thr Asp Cys Asn Ala Val Ala Val Val Glu Glu Ser Val Cys Lys Glu

195 200 205

Glu Glu Glu Val Ala Leu Val Val Gly Lys Gly Val Glu Glu Glu Glu

210 215 220

Ala Gly Met Ser Glu Arg Arg Lys Arg Met Thr Met Glu Val Phe Val

225 230 235 240

Gly Gly Leu His Arg Asp Ala Lys Glu Asp Asp Val Arg Ala Val Phe

245 250 255

Ala Lys Ala Gly Glu Ile Thr Glu Val Arg Met Ile Met Asn Pro Leu

260 265 270

Ala Gly Lys Asn Lys Gly Tyr Cys Phe Val Arg Tyr Arg His Ala Ala

275 280 285

Gln Ala Lys Lys Ala Ile Ala Glu Phe Gly Asn Val Lys Ile Cys Gly

290 295 300

Lys Leu Cys Arg Ala Ala Val Pro Val Gly Asn Asp Arg Ile Phe Leu

305 310 315 320

Gly Asn Ile Asn Lys Lys Trp Lys Lys Glu Asp Val Ile Lys Gln Leu

325 330 335

Lys Lys Ile Gly Ile Glu Asn Ile Asp Ser Val Thr Leu Lys Ser Asp

340 345 350

Ser Asn Asn Pro Val Cys Asn Arg Gly Phe Ala Phe Leu Glu Leu Glu

355 360 365

Thr Ser Arg Asp Ala Arg Met Ala Tyr Lys Lys Leu Ser Gln Lys Asn

370 375 380

Ala Phe Gly Lys Gly Leu Asn Ile Arg Val Ala Trp Ala Glu Pro Leu

385 390 395 400

Asn Asp Pro Asp Glu Lys Asp Met Gln Val Lys Ser Ile Phe Val Asp

405 410 415

Gly Ile Pro Thr Ser Trp Asp His Ala Gln Leu Lys Glu Ile Phe Lys

420 425 430

Lys His Gly Lys Ile Glu Ser Val Val Leu Ser Arg Asp Met Pro Ser

435 440 445

Ala Lys Arg Arg Asp Phe Ala Phe Ile Asn Tyr Ile Thr Arg Glu Ala

450 455 460

Ala Ile Ser Cys Leu Glu Ser Phe Asp Lys Glu Glu Phe Ser Lys Asn

465 470 475 480

Gly Ser Lys Val Asn Ile Lys Val Ser Leu Ala Lys Pro Ala Gln Gln

485 490 495

Ser Lys Gln Thr Lys Glu Asp His Lys Ser Ser Ile Thr Gly Glu Gly

500 505 510

Lys Met Lys Thr Ser Lys Ile Arg Tyr Pro Val Gln Asp Tyr Thr His

515 520 525

Ile Tyr Ser Gly Glu Lys Arg Pro Phe Ser Thr Leu Gly Asp Pro Tyr

530 535 540

Tyr Pro Leu Arg Gly His Ser Cys Arg Arg His Glu Gly Ser Thr Tyr

545 550 555 560

Thr Thr Ala Ala Ser Ser Tyr Gly Ala Leu Pro Pro Ala Thr Ala Glu

565 570 575

Ser Ser Leu Pro His Tyr His Asp Ser Asn Arg Tyr Pro Pro His Leu

580 585 590

Gly Glu Ala Ile Lys Phe Ser Pro Thr Ser Ala Val Leu Ser Lys Gln

595 600 605

Ala Trp Gln Lys Met

610

<210>11

<211>654

<212>DNA

＜213〉rice

<400>11

cttctacatc ggcttaggtg tagcaacacg actttattat tattattatt attattatta 60

ttattttaca aaaatataaa atagatcagt ccctcaccac aagtagagca agttggtgag 120

ttattgtaaa gttctacaaa gctaatttaa aagttattgc attaacttat ttcatattac 180

aaacaagagt gtcaatggaa caatgaaaac catatgacat actataattt tgtttttatt 240

attgaaatta tataattcaa agagaataaa tccacatagc cgtaaagttc tacatgtggt 300

gcattaccaa aatatatata gcttacaaaa catgacaagc ttagtttgaa aaattgcaat 360

ccttatcaca ttgacacata aagtgagtga tgagtcataa tattattttc tttgctaccc 420

atcatgtata tatgatagcc acaaagttac tttgatgatg atatcaaaga acatttttag 480

gtgcacctaa cagaatatcc aaataatatg actcacttag atcataatag agcatcaagt 540

aaaactaaca ctctaaagca accgatggga aagcatctat aaatagacaa gcacaatgaa 600

aatcctcatc atccttcacc acaattcaaa tattatagtt gaagcatagt agta 654

<210>12

<211>30

<212>PRT

＜213〉artificial sequence

<220>

＜223〉motif I-consensus sequence

<400>12

Pro Tyr Glu Ala Ala Val Val Ala Leu Pro Val Val Val Lys Glu Arg

1 5 10 15

Leu Val Arg Ile Leu Arg Leu Gly Ile Ala Thr Arg Tyr Asp

20 25 30

<210>13

<211>14

<212>PRT

＜213〉artificial sequence

<220>

＜223〉motif II-consensus sequence

<400>13

Arg Phe Asp Pro Phe Thr Gly Glu Pro Tyr Lys Phe Asp Pro

1 5 10

<210>14

<211>2166

<212>DNA

＜213〉Arabidopis thaliana (Arabidopsis thaliana)

<400>14

aagatttggg cttacaatct ttatcacaaa ggctttttta aagcccatta gttacattca 60

tcattatctc tcgacattaa aaaaaaaaag ttaaactgaa gaagctaaaa agagttttta 120

acttttaact ctcttcgtct tctccctcgt gccgtgtcaa atcaatctac tgttctctct 180

cctatctggt aaacttttcc tcttcgccat gaaatttttt tcttgctagg gttttagttt 240

ctacagttcg cttcccaaaa attaggggtt ttgtcacaat ttctcaattt cttgttccat 300

ttttcttctt ttctccataa tcattgctta atttagaatc ccaaatttta caaattaggg 360

tttttgttta attttagggg tttttgattt tcaactgtta atagtgttct cgatgtcata 420

attctgattt tttttattat ctattccgaa attagggcaa aaatctcaga caaacctgca 480

aaattagggt atttgaggat atggattatg atcggtacaa gttatttgtt ggtggtattg 540

cgaaagagac aagtgaagaa gctctgaagc agtattttag cagatatgga gctgtgttgg 600

aagctgttgt agctaaagag aaagtcactg gaaaacctag aggttttggg tttgttcgct 660

ttgctaatga ttgtgatgtt gttaaagctc ttagagacac tcacttcatt ctcggtaaac 720

ccgtaagtgt taccgccttt ttatgcttgt gtcaattggg ttttgtgtat actctgtgga 780

ttgattatgt gtgtgtttgt attaggttga tgtgagaaag gcgattagga aacatgaact 840

ataccaacag ccgtttagca tgcagttttt ggagagaaaa gtgcaacaga tgaatggtgg 900

tttgcgtgag atgtcgagta atggtgtgac cagtaggact aagaagatat ttgttggggg 960

tttgtcgtct aacacgactg aggaagagtt taagagttac tttgagaggt ttggtaggac 1020

tactgatgta gttgtgatgc atgacggtgt gactaacagg ccaaggggtt ttgggtttgt 1080

tacttatgat tcggaggact ctgttgaggt tgttatgcag agtaatttcc atgagttgag 1140

tgataaacgc gtggaagtga aacgggcaat acctaaagaa ggaatccaga gcaataacgg 1200

taatgctgtt aatattcctc cttcctacag cagctttcaa gcaacacctt atgtccctga 1260

gcaaaacgga tatgggatgg ttttacagtt tcctcctcct gtctttggtt atcatcacaa 1320

tgtccaagcc gttcaatatc cttatggtta ccaattcaca gcacaagtgg ctaacgtttc 1380

atggaacaat ccgattatgc aacccaccgg tttttactgt gctcctcctc atcctactcc 1440

tcctcccacc aacaatcttg gttatatcca atacatgaac gggtttgatc tttcgggtac 1500

gaacatttcc gggtacaatc ctctagcatg gcctgtaacg ggggatgcag ctggtgcgct 1560

aatacatcag tttgtagatt tgaagcttga tgtccacagt caagcccatc agagaatgaa 1620

tggaggtaac atgggaatac cattgcagaa tggtacatat atatgacagt tgcagaatga 1680

taaatgcaaa taggctcaca agggtagtga aattctttgg actcttttaa atggtttttt 1740

aggttcctca tctttcttca ttaactcttt ggtaaatgtg ttgggttggt ttggttacct 1800

tgtatattgt ttaggtattt gattttaacc ccaagactta tgtatcatat attactgcat 1860

ttgtaatata tcacactcat ttagttcatt ttgttgcttt tatggttttg ttgattttgt 1920

ggtttcgttg attaaattgg caatgatgtt ttaaattcat caaggaaaac aaagaaatag 1980

attgtcgatt aaacagtaga aaaaggaaat agttttgtag aaataggaac tgaatctgga 2040

aatctctaag aataccatat tgtagaaaga aaataaatct gagacgggag aaactatcga 2100

gcatccttga gctttaagtt ggagaaaccg ggtaagcgtt tgtgggattt tgttgtaaga 2160

ttgaac 2166

<210>15

<211>360

<212>PRT

＜213〉Arabidopis thaliana

<400>15

Met Asp Tyr Asp Arg Tyr Lys Leu Phe Val Gly Gly Ile Ala Lys Glu

1 5 10 15

Thr Ser Glu Glu Ala Leu Lys Gln Tyr Phe Ser Arg Tyr Gly Ala Val

20 25 30

Leu Glu Ala Val Val Ala Lys Glu Lys Val Thr Gly Lys Pro Arg Gly

35 40 45

Phe Gly Phe Val Arg Phe Ala Asn Asp Cys Asp Val Val Lys Ala Leu

50 55 60

Arg Asp Thr His Phe Ile Leu Gly Lys Pro Val Asp Val Arg Lys Ala

65 70 75 80

Ile Arg Lys His Glu Leu Tyr Gln Gln Pro Phe Ser Met Gln Phe Leu

85 90 95

Glu Arg Lys Val Gln Gln Met Asn Gly Gly Leu Arg Glu Met Ser Ser

100 105 110

Asn Gly Val Thr Ser Arg Thr Lys Lys Ile Phe Val Gly Gly Leu Ser

115 120 125

Ser Asn Thr Thr Glu Glu Glu Phe Lys Ser Tyr Phe Glu Arg Phe Gly

130 135 140

Arg Thr Thr Asp Val Val Val Met His Asp Gly Val Thr Asn Arg Pro

145 150 155 160

Arg Gly Phe Gly Phe Val Thr Tyr Asp Ser Glu Asp Ser Val Glu Val

165 170 175

Val Met Gln Ser Asn Phe His Glu Leu Ser Asp Lys Arg Val Glu Val

180 185 190

Lys Arg Ala Ile Pro Lys Glu Gly Ile Gln Ser Asn Asn Gly Asn Ala

195 200 205

Val Asn Ile Pro Pro Ser Tyr Ser Ser Phe Gln Ala Thr Pro Tyr Val

210 215 220

Pro Glu Gln Asn Gly Tyr Gly Met Val Leu Gln Phe Pro Pro Pro Val

225 230 235 240

Phe Gly Tyr His His Asn Val Gln Ala Val Gln Tyr Pro Tyr Gly Tyr

245 250 255

Gln Phe Thr Ala Gln Val Ala Asn Val Ser Trp Asn Asn Pro Ile Met

260 265 270

Gln Pro Thr Gly Phe Tyr Cys Ala Pro Pro His Pro Thr Pro Pro Pro

275 280 285

Thr Asn Asn Leu Gly Tyr Ile Gln Tyr Met Asn Gly Phe Asp Leu Ser

290 295 300

Gly Thr Asn Ile Ser Gly Tyr Asn Pro Leu Ala Trp Pro Val Thr Gly

305 310 315 320

Asp Ala Ala Gly Ala Leu Ile His Gln Phe Val Asp Leu Lys Leu Asp

325 330 335

Val His Ser Gln Ala His Gln Arg Met Asn Gly Gly Asn Met Gly Ile

340 345 350

Pro Leu Gln Asn Gly Thr Tyr Ile

355 360

<210>16

<211>3041

<212>DNA

＜213〉Arabidopis thaliana

<400>16

cttcattgag agagagatat agagagagaa aagagagaga ggccatattt tgataagaga 60

agaagaaccc ttatagagaa agagaaagag agagacagag agagtggatg gatgtcttat 120

agaatgaaca aaacatcctc tgtttctctt gtccttgtcc ctttttccag atcttaaggt 180

tttccacatt ttatcatctg ggtcctctcc ttaatggtga attctccatc tttacaagtt 240

tgatgttttt gttcatcaaa tctggcgttt ttttttctct tctaatatat attgtctctg 300

ctcattttcc gtttctcttc ccattgattg ttctgtttca tttctgtttt ttttttttca 360

atagttttga ttggatgctt tgatgatcca ttgtcagatt tgaagacact caattcctat 420

ttgatcgggg actagaattt ggattctgtt tcagacaaaa gtagatttcc ctgtctcttt 480

cccgtttgat tttcaataag atgaatccgg aggtaaaaca ttgaacaatt cttcataaat 540

ctcagaactt tgagcttttt tgaatcttaa aacacgatcg aagtaaaaaa tcgaattgtt 600

agatgaaatg ggcaatcgtc attttcgcaa atctgatccg tatttgtgag atcggattca 660

ttggatcgac tttggggttt tgcaggagca aaagatggaa tctgcatcgg atctgggcaa 720

gctcttcatt ggcgggattt catgggacac agatgaagaa cgactgcaag agtattttgg 780

caagtatgga gatttggttg aagctgtgat catgagagac cgtactaccg gacgtgcccg 840

tggctttggg tttatcgttt ttgcagatcc ttctgttgcc gagagagtca tcatggacaa 900

acacatcatt gatggccgca cggttagtat tcttggatcc attgcttgac aattcatcta 960

attatcagtc ttgagtaatc gagtgttcta aagtctcgat ctttctgtaa tgattctgtc 1020

ttagaggtct tattggtctc gctgctcgtt aatgagcaac ggattgttct ataatctcga 1080

tctttctgta ttcatgctct cttagagatc tgtttggtgt catccattaa tgagttttaa 1140

gcagcaacgt ttagatcttt ctgtaatcat gctcttttcg aaatcttctg ttgtcattag 1200

cttctggatt tgctgttact gttataactt gtgagaatgt gttgttgctt tgtgttgaag 1260

tggcaatgtt agtgttagat caatgagaaa agaatgaaag atcttttttt atttctttgt 1320

tgcaggtcga ggcgaagaaa gctgtcccgc gggatgatca gcaagtgcta aaacgacacg 1380

ccagtccaat gcaccttatc tcacctagcc atggtggtaa tggtggtgga gcacggacaa 1440

agaagatctt tgttggaggt ttaccgtcta gcattactga ggccgagttc aagaactact 1500

ttgatcagtt tggtacaatt gctgatgttg tggtaatgta tgatcataat acacagaggc 1560

caagaggctt tggcttcatc acttttgatt ccgaagagtc tgttgatatg gttctccaca 1620

agacctttca tgagctaaac ggaaaaatgg ttgaggttaa aagagcagtg ccaaaggagc 1680

tctcctcgac tactcctaac cgaagcccac ttattgggta tggtaacaac tatggagtag 1740

tccctaatag gtcttctgct aatagctact tcaatagttt tcctcctggt tataataata 1800

ataatctagg ctctgctggc cggtttagtc ctattggtag cggtagaaat gctttctcta 1860

gcttcgggct cggattgaat caagaactga atttgaattc aaactttgat ggaaacactc 1920

ttgggtatag ccggatccct ggcaaccaat acttcaacag tgcttcacca aaccgttaca 1980

actctccaat tgggtacaac agaggagact ctgcttacaa cccgagcaac agagacttgt 2040

ggggaaacag aagcgattcc tctggtccag gttggaactt gggagtttcg gttggtaaca 2100

acagaggaaa ctggggactt tcttctgtgg tgagcgataa caatggctat ggaagaagct 2160

atggggctgg ttctggactt tcggggttat cattcgcggg taatacaaac ggttttgatg 2220

gctctatagg ggaattgtat agaggcagct cagtttatag cgactcaaca tggcagcagt 2280

caatgcctca tcatcagtct tctaatgagt tagacggctt gtctcgctct tatggctttg 2340

gtattgacaa tgtaggctca gacccatcag ccaatgcctc agaaggatac tccggaaact 2400

acaatgtcgg aaatagacaa acacatagag gtacactcat cgatgtcaaa cttttttcct 2460

tttgcatctc atctgctaca tttatttttg cctgttgaaa agtaattaga ttgattaacg 2520

ttttcaggta ttgaagcata gaaagaaatc gacgaagaga agtgagaatt gtagatcaag 2580

aagaacagcc atttccgttg cagagtttga agagttgtta tttcgatatc aagtagagaa 2640

agaaaccaac tttcttcatc acagtgagtt tcttgttttg tttttttcgt cgttagcatc 2700

acaaacacaa aaaagagaag tttattttta ctttaaaaat tcttacataa gataagatca 2760

gattggtagc tgcaaagata caacatggat gataaaaaaa gatttggttt cgtctccata 2820

gcaataacca gagatcgttg attctcgatc actattcttt aggtttctct ccttcttctt 2880

ccatgatttc ttgatgttgt gtgctctgtt tgtaactcta attgttaaaa ttttttatgt 2940

tacagatttt ttttttcttt tggtttttaa actttggatt cgaattgttc atgggaactt 3000

ttggattttt ctattagcgt gagagaaaac acattgtgca a 3041

<210>17

<211>455

<212>PRT

＜213〉Arabidopis thaliana

<400>17

Met Asn Pro Glu Glu Gln Lys Met Glu Ser Ala Ser Asp Leu Gly Lys

1 5 10 15

Leu Phe Ile Gly Gly Ile Ser Trp Asp Thr Asp Glu Glu Arg Leu Gln

20 25 30

Glu Tyr Phe Gly Lys Tyr Gly Asp Leu Val Glu Ala Val Ile Met Arg

35 40 45

Asp Arg Thr Thr Gly Arg Ala Arg Gly Phe Gly Phe Ile Val Phe Ala

50 55 60

Asp Pro Ser Val Ala Glu Arg Val Ile Met Asp Lys His Ile Ile Asp

65 70 75 80

Gly Arg Thr Val Glu Ala Lys Lys Ala Val Pro Arg Asp Asp Gln Gln

85 90 95

Val Leu Lys Arg His Ala Ser Pro Met His Leu Ile Ser Pro Ser His

100 105 110

Gly Gly Asn Gly Gly Gly Ala Arg Thr Lys Lys Ile Phe Val Gly Gly

115 120 125

Leu Pro Ser Ser Ile Thr Glu Ala Glu Phe Lys Asn Tyr Phe Asp Gln

130 135 140

Phe Gly Thr Ile Ala Asp Val Val Val Met Tyr Asp His Asn Thr Gln

145 150 155 160

Arg Pro Arg Gly Phe Gly Phe Ile Thr Phe Asp Ser Glu Glu Ser Val

165 170 175

Asp Met Val Leu His Lys Thr Phe His Glu Leu Asn Gly Lys Met Val

180 185 190

Glu Val Lys Arg Ala Val Pro Lys Glu Leu Ser Ser Thr Thr Pro Asn

195 200 205

Arg Ser Pro Leu Ile Gly Tyr Gly Asn Asn Tyr Gly Val Val Pro Asn

210 215 220

Arg Ser Ser Ala Asn Ser Tyr Phe Asn Ser Phe Pro Pro Gly Tyr Asn

225 230 235 240

Asn Asn Asn Leu Gly Ser Ala Gly Arg Phe Ser Pro Ile Gly Ser Gly

245 250 255

Arg Asn Ala Phe Ser Ser Phe Gly Leu Gly Leu Asn Gln Glu Leu Asn

260 265 270

Leu Asn Ser Asn Phe Asp Gly Asn Thr Leu Gly Tyr Ser Arg Ile Pro

275 280 285

Gly Asn Gln Tyr Phe Asn Ser Ala Ser Pro Asn Arg Tyr Asn Ser Pro

290 295 300

Ile Gly Tyr Asn Arg Gly Asp Ser Ala Tyr Asn Pro Ser Asn Arg Asp

305 310 315 320

Leu Trp Gly Asn Arg Ser Asp Ser Ser Gly Pro Gly Trp Asn Leu Gly

325 330 335

Val Ser Val Gly Asn Asn Arg Gly Asn Trp Gly Leu Ser Ser Val Val

340 345 350

Ser Asp Asn Asn Gly Tyr Gly Arg Ser Tyr Gly Ala Gly Ser Gly Leu

355 360 365

Ser Gly Leu Ser Phe Ala Gly Asn Thr Asn Gly Phe Asp Gly Ser Ile

370 375 380

Gly Glu Leu Tyr Arg Gly Ser Ser Val Tyr Ser Asp Ser Thr Trp Gln

385 390 395 400

Gln Ser Met Pro His His Gln Ser Ser Asn Glu Leu Asp Gly Leu Ser

405 410 415

Arg Ser Tyr Gly Phe Gly Ile Asp Asn Val Gly Ser Asp Pro Ser Ala

420 425 430

Asn Ala Ser Glu Gly Tyr Ser Gly Asn Tyr Asn Val Gly Asn Arg Gln

435 440 445

Thr His Arg Gly Ile Glu Ala

450 455

<210>18

<211>2524

<212>DNA

＜213〉Arabidopis thaliana

<400>18

atatgtgaga ctaactattg ttctctgtct ctttttttct ttttaattat caaagaaaga 60

aactctttct taatggaaac catttacaga taaaaaaaac attaaaagga aaggttttta 120

ataaagcctt tgagagagaa gatgtttatt ataggatgaa caaaaacatc ctctgtttct 180

ctcttttcat atttttctcc acatttcctc atctgggtca tctccaaaaa tggtgctttt 240

ttttaataat tcttcacgtt tctgggtttt tggttttgtg atttgatgat gctttttttt 300

tgtttttttc agatttgatg ataacccaaa ttcgcaattt gattaggaca acaacaacaa 360

ctttatttat ctgattccgt ctttgatttt cagacaagaa aagtatgttg tttctaagtc 420

ttttgatttt tttcaatttc atctccttac tcgatttttt tttttttggg tttctctgaa 480

ttggagcaga aaaaaaaaag atggaatcgg atctggggaa gctcttcatt ggtgggattt 540

cgtgggatac agacgaagaa aggttaagag actactttag caactatggt gatgttgttg 600

aagctgtgat catgagagat cgtgccacag gtcgtgcacg tggcttcggc ttcattgtct 660

ttgcagaccc ctgtgtctca gagagagtga tcatggataa acacatcatc gatggccgca 720

cggtttgtga tttcaatcat ttctcaatct ttcagcagaa caaacaaagt tcagatctta 780

ttgcaacttc ctcaatttgc gtttttgaat catctctcaa tctttgtttc tcaaagtgta 840

aagatcaaat ttatgttttg caggttgagg cgaagaaggc tgtgcctcga gatgatcagc 900

aggtgctaaa gcgacacgct agtcctatcc accttatgtc acctgtccat ggtggtggtg 960

gaaggacaaa gaagatcttc gttggaggtt taccgtctag cattaccgag gaggagttca 1020

agaactactt tgatcagttt ggtactattg ctgatgttgt tgtaatgtat gatcataaca 1080

cgcagaggcc aagaggtttt ggcttcatca catttgattc agatgatgct gttgatagag 1140

ttcttcacaa gaccttccat gagctcaatg ggaaactagt tgaggtcaaa agagctgtac 1200

ctaaggagat ttcccctgtt tctaatatcc gaagcccgct tgctagcggt gttaactatg 1260

gaggcgggtc taataggatg cctgctaata gctactttaa caactttgct cctggtcctg 1320

gtttttataa cagtctaggt cctgttggtc gtcggtttag tcctgttatt ggtagtggta 1380

gaaatgcggt ttctgctttt ggcctcggtt tgaatcatga cttgagtttg aatttgaatc 1440

caagctgcga tgggacaagt tctacgtttg gttataaccg tattccaagc aacccttact 1500

tcaacggtgc ttccccgaac cgttacacct ctccaatcgg gcacaataga actgagtctc 1560

cttacaattc gaacaataga gacttatggg gaaacagaac cgacactgca ggtcccggtt 1620

ggaacttgaa tgtctcgaat ggaaacaaca gaggaaattg gggacttcct tcttcttctg 1680

ctgttagtaa tgataacaat ggctttggaa ggaactatgg gacaagttct ggactttcct 1740

cgtccccatt taatggtttt gaaggttcta taggggaact gtacagaggc ggctcagtct 1800

acagcgactc aacgtggcag caacagcagc taccatctca gtcttctcac gagctagaca 1860

atttgtctcg cgcttacggt tatgatattg acaatgtagg ttcagaccca tctgcaaatg 1920

acccagaaac ttacaatgga agctacaatg ttggaaatag acaaactaat agaggtaaca 1980

aaaaaattca tctcaataaa acttgtaact tggatacatt ttgatcgcaa tcgaaatgtt 2040

ctgatctgtg ttttatttac ttgttgaggt attgctgcat aggttatcaa aaaccaagaa 2100

aacaaaaaaa aaagttgaga gatttgtaga ttgaaagcaa ccaaatttca gttgcagagt 2160

ttgaacaggt tctcatgaca aagaaaccaa ctttgttgat cacagtgcca aagattatgg 2220

tttgctttct cttttgttag accaaaaaaa aaaaaaaaaa agagaaaaac aaagaaccgt 2280

ttttgttttt cttcttctta cataaagatc agatcgtagc agccagacaa ccaaagatac 2340

tacaaggtgg atttagattt gcttctcaaa aaagtttttt tttttctttc atagaataac 2400

caaacaaaga tcgtagaatt ttcgatcaaa gattcttcag agttctgtgc tctgttttgt 2460

aattgtactt tttttttctt gtttacaaaa tgaattgttc atgaaaactt tgttttctta 2520

aaaa 2524

<210>19

<211>460

<212>PRT

＜213〉Arabidopis thaliana

<400>19

Met Glu Ser Asp Leu Gly Lys Leu Phe Ile Gly Gly Ile Ser Trp Asp

1 5 10 15

Thr Asp Glu Glu Arg Leu Arg Asp Tyr Phe Ser Asn Tyr Gly Asp Val

20 25 30

Val Glu Ala Val Ile Met Arg Asp Arg Ala Thr Gly Arg Ala Arg Gly

35 40 45

Phe Gly Phe Ile Val Phe Ala Asp Pro Cys Val Ser Glu Arg Val Ile

50 55 60

Met Asp Lys His Ile Ile Asp Gly Arg Thr Val Glu Ala Lys Lys Ala

65 70 75 80

Val Pro Arg Asp Asp Gln Gln Val Leu Lys Arg His Ala Ser Pro Ile

85 90 95

His Leu Met Ser Pro Val His Gly Gly Gly Gly Arg Thr Lys Lys Ile

100 105 110

Phe Val Gly Gly Leu Pro Ser Ser Ile Thr Glu Glu Glu Phe Lys Asn

115 120 125

Tyr Phe Asp Gln Phe Gly Thr Ile Ala Asp Val Val Val Met Tyr Asp

130 135 140

His Asn Thr Gln Arg Pro Arg Gly Phe Gly Phe Ile Thr Phe Asp Ser

145 150 155 160

Asp Asp Ala Val Asp Arg Val Leu His Lys Thr Phe His Glu Leu Asn

165 170 175

Gly Lys Leu Val Glu Val Lys Arg Ala Val Pro Lys Glu Ile Ser Pro

180 185 190

Val Ser Asn Ile Arg Ser Pro Leu Ala Ser Gly Val Asn Tyr Gly Gly

195 200 205

Gly Ser Asn Arg Met Pro Ala Asn Ser Tyr Phe Asn Asn Phe Ala Pro

210 215 220

Gly Pro Gly Phe Tyr Asn Ser Leu Gly Pro Val Gly Arg Arg Phe Ser

225 230 235 240

Pro Val Ile Gly Ser Gly Arg Asn Ala Val Ser Ala Phe Gly Leu Gly

245 250 255

Leu Asn His Asp Leu Ser Leu Asn Leu Asn Pro Ser Cys Asp Gly Thr

260 265 270

Ser Ser Thr Phe Gly Tyr Asn Arg Ile Pro Ser Asn Pro Tyr Phe Asn

275 280 285

Gly Ala Ser Pro Asn Arg Tyr Thr Ser Pro Ile Gly His Asn Arg Thr

290 295 300

Glu Ser Pro Tyr Asn Ser Asn Asn Arg Asp Leu Trp Gly Asn Arg Thr

305 310 315 320

Asp Thr Ala Gly Pro Gly Trp Asn Leu Asn Val Ser Asn Gly Asn Asn

325 330 335

Arg Gly Asn Trp Gly Leu Pro Ser Ser Ser Ala Val Ser Asn Asp Asn

340 345 350

Asn Gly Phe Gly Arg Asn Tyr Gly Thr Ser Ser Gly Leu Ser Ser Ser

355 360 365

Pro Phe Asn Gly Phe Glu Gly Ser Ile Gly Glu Leu Tyr Arg Gly Gly

370 375 380

Ser Val Tyr Ser Asp Ser Thr Trp Gln Gln Gln Gln Leu Pro Ser Gln

385 390 395 400

Ser Ser His Glu Leu Asp Asn Leu Ser Arg Ala Tyr Gly Tyr Asp Ile

405 410 415

Asp Asn Val Gly Ser Asp Pro Ser Ala Asn Asp Pro Glu Thr Tyr Asn

420 425 430

Gly Ser Tyr Asn Val Gly Asn Arg Gln Thr Asn Arg Gly Asn Lys Lys

435 440 445

Ile His Leu Asn Lys Thr Cys Asn Leu Asp Thr Phe

450 455 460

<210>20

<211>2607

<212>DNA

＜213〉Arabidopis thaliana

<400>20

ctgtaatgtg gagtttggaa ttttcgacaa caaagtgcac atctggcaca gagattgtca 60

cagcacgaaa gatttttttg tcgttcttgt aggatttgct ggcacgtgtg gaatagaaaa 120

cacacgagtg aaaccatcgt cggtctttgt agcccattat ttatacttct attgggctgg 180

acttaagccc ataagtaagc atctctgtta caagaaaacg ggaaacagat ctgaaccgtt 240

aataatatta gaaaggatct agaccgttga tttatttatc tgctgacaga ttcgtacctt 300

cgcgaatatc aataccaaac caatagaaat attcgttcgc tgtcttcttc ctcttcctcc 360

tctcaaatcg gctacagcca ttggaaaagc taaagccttt tcgtaatttc tggaagtttc 420

tgcagtcggt tttcacggtt tcgtagattg aggtggattt gtgattctgg gtcagaagta 480

agatagtgga atataaattc atggattcgg atcaaggaaa gctttttgtc ggtggtattt 540

catgggaaac tgatgaagat aagctgagag aacatttcac caactatgga gaggtttctc 600

aggctattgt gatgagagac aagctcacag gtcgacctag gggttttggg ttcgttatct 660

tctcggatcc ttctgttctc gatagggttc ttcaagagaa acacagcatt gataccagag 720

aggttattat tgttctctta tagctccatt tctctaattg tgttaaagtt ttatcctttt 780

tgcgttttgc tgtgttgatt gagaacgaga gtaaatatag aattttgttt ggttggcaaa 840

ttcgccttag tgtttcttag attctaggat tggttttaac ttgtataaga ggtattatag 900

ggtactcgat atatgttaat cgtacactct atgaagtgat tgagtatagt attagaaaag 960

agagcttggt ttggtttatt aggataagga aaaacagatg tatatatttt ctgttgcgtt 1020

atgttctcga tttgggtaaa gtatgattct tggaagttta ttatgagctt tattgatttt 1080

ggttaatgtt taggttgatg tgaagagagc catgtcaaga gaggagcagc aagtctctgg 1140

aagaactggg aatcttaata catctagaag ttctggaggt gatgcttaca ataaaaccaa 1200

gaagatcttt gttggaggct tgccacctac tttgactgat gaagagtttc gccagtactt 1260

tgaagtttat ggccctgtga ctgatgttgc aatcatgtat gaccaggcta ccaaccgtcc 1320

tcgtgggttt ggatttgttt ccttcgactc tgaagatgcg gtagacagtg ttttgcacaa 1380

gactttccat gatttgagcg gtaaacaagt tgaagtaaag cgtgctcttc ctaaagatgc 1440

caatcctgga ggtggtggac gatcaatggg tggtggtggc tctggtggtt accagggtta 1500

tggtggcaat gaaagcagtt atgatggacg tatggattcc aataggtttt tgcagcatca 1560

aagtgttgga aatggtttac catcttatgg ttcttctggt tatggcgctg gctatggaaa 1620

tggtagtaat ggtgccgggt atggtgccta tggaggttac actggttctg ctggaggtta 1680

tggcgctggt gctactgctg gatatggagc aacgaacatt ccaggtgctg gctatggaag 1740

tagtactgga gttgctccga gaaactcatg ggacactcca gcttctagtg gttatgggaa 1800

cccaggctat gggagtggtg ctgctcatag tggatatgga gttcctggtg cagctcctcc 1860

tacgcagtca ccatctggct atagtaacca aggctacggt tatggagggt acagtggaag 1920

tgattctggt tatggaaatc aagctgcata tggtgtggtt ggagggcgtc ctagtggtgg 1980

cggttcaaac aaccctggta gtggtggcta catgggaggt ggttatggtg atggatcttg 2040

gcgatctgac ccgtcacaag gttatggtgg tgggtacaat gatggtcagg gtcgacaagg 2100

ccagtagtga ctgtgtaagg ggattatgac cgccctggtt tctggatcct tgtcaagaag 2160

aatttagctc aaatcaaagg ttccacaact tcctaacggg ttggactgct tgaatctctt 2220

tataagcatg tgctatctat tacaataagt cacttctatt aagttatttt tcggttgagt 2280

gtacttttga gttttggcag agttattata actacaggct ttgctgtttt cgtattatgt 2340

ttgtcttcct agtattcttg ccggattgtt tgttttgatt gtgttatttt gttttggccc 2400

tgatggatat aacttaagca gggaataatg cttcagggta cttgttaaga aagcagatgg 2460

tgagagcaga actcgatgga ggtgagagtc aaattgctga atgtatggtt tgagtagaaa 2520

gtagaggtag ttggtaacgt tagtggtacc attaagaaga aggtgtagaa aatagtgaga 2580

ggtagctttg agaaaaaggc ataatca 2607

<210>21

<211>406

<212>PRT

＜213〉Arabidopis thaliana

<400>21

Met Asp Ser Asp Gln Gly Lys Leu Phe Val Gly Gly Ile Ser Trp Glu

1 5 10 15

Thr Asp Glu Asp Lys Leu Arg Glu His Phe Thr Asn Tyr Gly Glu Val

20 25 30

Ser Gln Ala Ile Val Met Arg Asp Lys Leu Thr Gly Arg Pro Arg Gly

35 40 45

Phe Gly Ile Arg Lys Asn Arg Cys Ile Tyr Phe Leu Leu Arg Tyr Val

50 55 60

Leu Asp Leu Gly Lys Val Asp Val Lys Arg Ala Met Ser Arg Glu Glu

65 70 75 80

Gln Gln Val Ser Gly Arg Thr Gly Asn Leu Asn Thr Ser Arg Ser Ser

85 90 95

Gly Gly Asp Ala Tyr Asn Lys Thr Lys Lys Ile Phe Val Gly Gly Leu

100 105 110

Pro Pro Thr Leu Thr Asp Glu Glu Phe Arg Gln Tyr Phe Glu Val Tyr

115 120 125

Gly Pro Val Thr Asp Val Ala Ile Met Tyr Asp Gln Ala Thr Asn Arg

130 135 140

Pro Arg Gly Phe Gly Phe Val Ser Phe Asp Ser Glu Asp Ala Val Asp

145 150 155 160

Ser Val Leu His Lys Thr Phe His Asp Leu Ser Gly Lys Gln Val Glu

165 170 175

Val Lys Arg Ala Leu Pro Lys Asp Ala Asn Pro Gly Gly Gly Gly Arg

180 185 190

Ser Met Gly Gly Gly Gly Ser Gly Gly Tyr Gln Gly Tyr Gly Gly Asn

195 200 205

Glu Ser Ser Tyr Asp Gly Arg Met Asp Ser Asn Arg Phe Leu Gln His

210 215 220

Gln Ser Val Gly Asn Gly Leu Pro Ser Tyr Gly Ser Ser Gly Tyr Gly

225 230 235 240

Ala Gly Tyr Gly Asn Gly Ser Asn Gly Ala Gly Tyr Gly Ala Tyr Gly

245 250 255

Gly Tyr Thr Gly Ser Ala Gly Gly Tyr Gly Ala Gly Ala Thr Ala Gly

260 265 270

Tyr Gly Ala Thr Asn Ile Pro Gly Ala Gly Tyr Gly Ser Ser Thr Gly

275 280 285

Val Ala Pro Arg Asn Ser Trp Asp Thr Pro Ala Ser Ser Gly Tyr Gly

290 295 300

Asn Pro Gly Tyr Gly Ser Gly Ala Ala His Ser Gly Tyr Gly Val Pro

305 310 315 320

Gly Ala Ala Pro Pro Thr Gln Ser Pro Ser Gly Tyr Ser Asn Gln Gly

325 330 335

Tyr Gly Tyr Gly Gly Tyr Ser Gly Ser Asp Ser Gly Tyr Gly Asn Gln

340 345 350

Ala Ala Tyr Gly Val Val Gly Gly Arg Pro Ser Gly Gly Gly Ser Asn

355 360 365

Asn Pro Gly Ser Gly Gly Tyr Met Gly Gly Gly Tyr Gly Asp Gly Ser

370 375 380

Trp Arg Ser Asp Pro Ser Gln Gly Tyr Gly Gly Gly Tyr Asn Asp Gly

385 390 395 400

Gln Gly Arg Gln Gly Gln

405

<210>22

<211>3178

<212>DNA

＜213〉Arabidopis thaliana

<400>22

ttgaaattgg gttaaatcgg tttgaatcgg attgaacaaa aactgtatta ataataattc 60

ttcctctact tttctctctg attgattcca atcttctttc attttcttct tcttcttctt 120

ctggggaagg ggcaggttaa aattatgcca tctattcaaa tcgtgcctat cctcagatct 180

taactctttt ctctacgaga ttcggcatct gggttttatt cttcttggtg ggtttttttt 240

tattcttctt cttctgatct cagatttccc ctgattggtt tttttttttg ctaaatccgt 300

tttatgtttt cccgatcaaa ctctcctggc agattctcgg atctgttgtt ttctagattc 360

aatctgaatt tgattttacg tttttgtctt tgtaaagatg tttccttttg atcagatttt 420

gataatccat tgacatctct gattcaagca aaagctaatt aactttgatc cgattccttt 480

gtgtgtgtgt gcagagcaaa atgcaatcgg ataatggaaa gcttttcatc ggtgggatat 540

cttgggacac caatgaggaa cgtctcaagg agtatttcag cagttttgga gaagtgatcg 600

aagctgtcat cttgaaagat cgtaccactg gtcgtgctcg tggtttcggt tttgttgttt 660

ttgctgatcc tgctgttgct gagattgtta tcaccgaaaa acataatatt gatggcagat 720

tggtatgttc actgttctct gcctttcgtt tttgtacaat gtaacttgtt ttcgaagctt 780

ccttatgcaa tcaagccttc aagagttaca gtttgttctc atttggttcc gattaatcat 840

ttttgtgctt tgattggatt tttgagaaga aatgagtgat ctttagttat atgagcttag 900

tttttcattt ttcaagttgt ttgatcttcc gcaggttgaa gccaagaaag ctgttcccag 960

agatgaccaa aacatggtaa atagaagcaa cagcagtagc atccaaggtt ctcccggtgg 1020

tccaggtcgc acaaggaaga tatttgttgg aggattacct tcttcggtta cagagagtga 1080

tttcaagacg tattttgagc agtttggtac aactacggat gtggttgtca tgtatgatca 1140

caacacacaa aggcctagag gtttcgggtt tataacctac gattccgagg aggcggttga 1200

aaaggtattg ctcaagacat tccatgaact aaatggtaaa atggttgagg ttaagcgagc 1260

tgttccaaag gagttatctc caggtccaag tcgcagtcct cttggtgcag gttacagcta 1320

tggagttaat agggtcaata acctccttaa tgggtatgct caagggttta atcccgctgc 1380

agttggaggc tacggactta ggatggatgg tcggttcagt ccggttggtg ctggaagaag 1440

cgggtttgca aattacagtt ctggatacgg gatgaatgtg aactttgatc agggattgcc 1500

cacagggttc acgggaggta caaattacaa tggaaatgtt gactatggcc gaggaatgag 1560

cccgtactac attggtaaca caaacaggtt tggtcctgcg gttggctatg aagggggcaa 1620

cggaggagga aactcatcct tcttcagttc ggttacacgg aacttatggg gaaacaatgg 1680

tggtcttaac tataacaaca ataatacaaa ctcaaactcc aatacatata tgggaggatc 1740

atcaagtggg aacaacacac ttagtggtcc atttggaaat tcaggagtca attggggtgc 1800

tcctggagga ggaaacaatg ctgtgagtaa cgagaatgtg aagtttggtt atggaggaaa 1860

cggtgaatct ggttttgggt tgggaacagg tggttatgca gcaagaaacc caggggctaa 1920

caaggcagca ccatcctctt cattctcttc tgcctcagca accaacaaca cgggttatga 1980

tacagcagga cttgcagagt tttacgggaa tggtgcagtt tatagtgacc ctacatggag 2040

atcaccaact cctgagacag aagggcctgc tccttttagc tatgggattg gaggaggggt 2100

tccttcttca gatgtttcag ctagaagttc atctccaggt tatgttggca gttacagtgt 2160

gaacaagaga caaccaaaca gaggtaattg agttcagagt aattttctgc tttaacatgt 2220

gattctatga aaagcaaagg actcttgaga aaaagaattt agaaagccta gatagtttcc 2280

aaatttttga ttatcctcgt cttctttctg gaatatacaa accatggttt agggtcttgc 2340

actaatggtg atctagaaca ccttcgtatc actagtgaat tggcttttcc tcagaaacac 2400

gaatatactt gcatgcagaa acagtagcca ttctgcatct ttattgtttt ttagttcatc 2460

agagattatt tagaggaaag tttctttccg tgctttagat ataagctcat ggaactagaa 2520

aactagttga atcttttatg ttgctcacac cagtgtctat gggaagtcta agaaacttgt 2580

gatgaagaaa ctcaattgca tgactggttt cttatcgctc ttctcttctc tgaattatat 2640

ttcccttttt cggttttgtt gcaggaattg ctacttagta caatcgtttt tgttttacca 2700

cgatattgta ggcgagccat cacggtgaac gatctgtgtc ttttggcgaa tcttttagat 2760

tatcttcttt tcccttcata caaagccagt gaggacgaaa cttgatcata tcatcaccta 2820

gagctaacca gagaatcccg cagacttttc tgtcatggtt tggttttcta aattcattgt 2880

tcctcctagg cttttttctg ctttcttttt ttttctattt ttgttttctt ttcttctttc 2940

aatgagggac agaagaaact gtatcagtct ccggcgaggc ggtaatacat aaggagagtt 3000

caaaacaaaa acccaaaaaa aaaaaaaaaa agatgatcct tcttcctcag ttttcttctt 3060

cattgtcatg taatggttct tcttcttttc ttcttcttgg gggttatggt taaggtttgt 3120

gttttgaggc agattgtact agagtttttt ttcatgtttc ttttgttttg tcgttttt 3178

<210>23

<211>494

<212>PRT

＜213〉Arabidopis thaliana

<400>23

Met Gln Ser Asp Asn Gly Lys Leu Phe Ile Gly Gly Ile Ser Trp Asp

1 5 10 15

Thr Asn Glu Glu Arg Leu Lys Glu Tyr Phe Ser Ser Phe Gly Glu Val

20 25 30

Ile Glu Ala Val Ile Leu Lys Asp Arg Thr Thr Gly Arg Ala Arg Gly

35 40 45

Phe Gly Phe Val Val Phe Ala Asp Pro Ala Val Ala Glu Ile Val Ile

50 55 60

Thr Glu Lys His Asn Ile Asp Gly Arg Leu Val Glu Ala Lys Lys Ala

65 70 75 80

Val Pro Arg Asp Asp Gln Asn Met Val Asn Arg Ser Asn Ser Ser Ser

85 90 95

Ile Gln Gly Ser Pro Gly Gly Pro Gly Arg Thr Arg Lys Ile Phe Val

100 105 110

Gly Gly Leu Pro Ser Ser Val Thr Glu Ser Asp Phe Lys Thr Tyr Phe

115 120 125

Glu Gln Phe Gly Thr Thr Thr Asp Val Val Val Met Tyr Asp His Asn

130 135 140

Thr Gln Arg Pro Arg Gly Phe Gly Phe Ile Thr Tyr Asp Ser Glu Glu

145 150 155 160

Ala Val Glu Lys Val Leu Leu Lys Thr Phe His Glu Leu Asn Gly Lys

165 170 175

Met Val Glu Val Lys Arg Ala Val Pro Lys Glu Leu Ser Pro Gly Pro

180 185 190

Ser Arg Ser Pro Leu Gly Ala Gly Tyr Ser Tyr Gly Val Asn Arg Val

195 200 205

Asn Asn Leu Leu Asn Gly Tyr Ala Gln Gly Phe Asn Pro Ala Ala Val

210 215 220

Gly Gly Tyr Gly Leu Arg Met Asp Gly Arg Phe Ser Pro Val Gly Ala

225 230 235 240

Gly Arg Ser Gly Phe Ala Asn Tyr Ser Ser Gly Tyr Gly Met Asn Val

245 250 255

Asn Phe Asp Gln Gly Leu Pro Thr Gly Phe Thr Gly Gly Thr Asn Tyr

260 265 270

Asn Gly Asn Val Asp Tyr Gly Arg Gly Met Ser Pro Tyr Tyr Ile Gly

275 280 285

Asn Thr Asn Arg Phe Gly Pro Ala Val Gly Tyr Glu Gly Gly Asn Gly

290 295 300

Gly Gly Asn Ser Ser Phe Phe Ser Ser Val Thr Arg Asn Leu Trp Gly

305 310 315 320

Asn Asn Gly Gly Leu Asn Tyr Asn Asn Asn Asn Thr Asn Ser Asn Ser

325 330 335

Asn Thr Tyr Met Gly Gly Ser Ser Ser Gly Asn Asn Thr Leu Ser Gly

340 345 350

Pro Phe Gly Asn Ser Gly Val Asn Trp Gly Ala Pro Gly Gly Gly Asn

355 360 365

Asn Ala Val Ser Asn Glu Asn Val Lys Phe Gly Tyr Gly Gly Asn Gly

370 375 380

Glu Ser Gly Phe Gly Leu Gly Thr Gly Gly Tyr Ala Ala Arg Asn Pro

385 390 395 400

Gly Ala Asn Lys Ala Ala Pro Ser Ser Ser Phe Ser Ser Ala Ser Ala

405 410 415

Thr Asn Asn Thr Gly Tyr Asp Thr Ala Gly Leu Ala Glu Phe Tyr Gly

420 425 430

Asn Gly Ala Val Tyr Ser Asp Pro Thr Trp Arg Ser Pro Thr Pro Glu

435 440 445

Thr Glu Gly Pro Ala Pro Phe Ser Tyr Gly Ile Gly Gly Gly Val Pro

450 455 460

Ser Ser Asp Val Ser Ala Arg Ser Ser Ser Pro Gly Tyr Val Gly Ser

465 470 475 480

Tyr Ser Val Asn Lys Arg Gln Pro Asn Arg Gly Ile Ala Thr

485 490

<210>24

<211>2351

<212>DNA

＜213〉Arabidopis thaliana

<400>24

atgatctaac attttttctc aaataataag gtcattgatc cttatataac atggaatcac 60

tataacattt ataacctaca ttcttgctca tatatctctc tccttttttt tccaacatat 120

taacgactaa taataaaatt tatcaaccat tttaaatctc taaatggaac ttattattac 180

atgactaaaa aataaaaata aataaataaa taaacgaagc tgatatggaa aagtcttctc 240

tttctttttt tttttttggt aagtcgatct ctctttcact cactttaacc caattggccg 300

ctattttcca aagtctgttt atttttttaa tctctctctc ttctctctca cccaatttca 360

caaacccgaa accctaattt tctcgggaca ctgaaatttt tacagcttct ttcctcttct 420

tcaccgggga gatttgtcgg tactaaatct agggtttttg ggtatcaccg gagggttgaa 480

gagagagaaa aaaactcaca atggaatcag atcagggaaa gctatttatc ggcgggattt 540

catgggatac cgacgagaat cttctgagag agtacttcag caatttcggc gaggttttgc 600

aggtcactgt tatgcgagag aaagctactg gtcgtcctag aggattcgga ttcgtcgcat 660

tctcggatcc tgctgttatt gatagggttc ttcaggacaa gcaccatatt gataatagag 720

atgtaagcaa aaatcttgtt tctcaaatgg gtctttctaa attttgaatc tttatagtaa 780

aaattgatac tttgaatctt gttgttgtcg aggtttgatt ttcatctttg atggatttaa 840

gttgtgttaa tttcttaggt tgatgtgaag agagcaatgt ctagagagga gcagagtcct 900

gctgggagat cagggacttt taatgcttct aggaattttg atagtggagc taacgtgagg 960

actaagaaga tattcgtggg aggtttgcct cctgcattaa catcagatga atttcgggct 1020

tactttgaga cttatggtcc tgtgagtgat gcagtcatta tgattgatca gactacacag 1080

cgtcctcgag gatttgggtt tgtttctttt gattctgaag attcggttga ccttgtttta 1140

cataagactt tccacgattt gaatggtaaa caagtcgaag ttaaaagagc tcttcctaaa 1200

gatgctaacc ctggaatagc cagtggtggt ggtcgtggca gtggtggagc tggagggttt 1260

ccgggctatg gtggttctgg tggaagtggc tatgagggtc gtgtggattc taatagatac 1320

atgcagccgc aaaacactgg aagtggttat cctccttatg gtggttctgg gtatggtact 1380

ggttatggtt atggaagcaa tggtgtaggt tatgggggtt ttggtgggta tggcaatcca 1440

gctggtgcgc cttatgggaa tcctagtgtc cctggagctg ggtttggaag tggtccaaga 1500

agttcatggg gcgctcaagc accatcgggt tatgggaatg tgggatatgg aaatgcagct 1560

ccgtggggtg gttctggtgg tcctggttca gcagtaatgg gtcaagctgg tgcatctgca 1620

ggttatggca gtcaaggtta tggctatggt ggaaatgatt cctcttacgg gactccatct 1680

gcctatggtg cagtaggggg gcgatctggg aatatgccta acaaccatgg tggcggtggc 1740

tatgcggatg ctttagatgg ctctggaggc tatgggaatc accaagggaa caacgggcaa 1800

gctggttatg gtggaggtta tggaagtggt aggcaagctc aacaacagtg attgaagaag 1860

aaatactact agaatgtggt tttatcgctg accttgaaac ctcctgcttt ccgccttaac 1920

catgtcacgt ctttggcggt tagaccagga ggtggaccta cgctggatta tctcttttgt 1980

tagtttctca ataagttgtt ttcaggcaat tccggatact atttcctatc aagttgtagt 2040

ttttaagttt gcgtgcttat ttatatttgt cgctttggaa tggttttctt tctctgttat 2100

cctctagtgt ttgtgtttaa cgatacatcc tccagattat cattattcat ctcccttttg 2160

gttcattcat ttttgttgaa tattccattc acagattctt gcttttgcat ctcctctgtt 2220

taggggaaga tgatttgctc agtgttcaat gtgatctaag aaaagtgttt ggtagagcaa 2280

gagctgcaat aaatcacttt gagattgcgt tgttacatga aggtcgtgtt ggcggaaact 2340

taacagtccc a 2351

<210>25

<211>404

<212>PRT

＜213〉Arabidopis thaliana

<400>25

Met Glu Ser Asp Gln Gly Lys Leu Phe Ile Gly Gly Ile Ser Trp Asp

1 5 10 15

Thr Asp Glu Asn Leu Leu Arg Glu Tyr Phe Ser Asn Phe Gly Glu Val

20 25 30

Leu Gln Val Thr Val Met Arg Glu Lys Ala Thr Gly Arg Pro Arg Gly

35 40 45

Phe Gly Phe Val Ala Phe Ser Asp Pro Ala Val Ile Asp Arg Val Leu

50 55 60

Gln Asp Lys His His Ile Asp Asn Arg Asp Val Asp Val Lys Arg Ala

65 70 75 80

Met Ser Arg Glu Glu Gln Ser Pro Ala Gly Arg Ser Gly Thr Phe Asn

85 90 95

Ala Ser Arg Asn Phe Asp Ser Gly Ala Asn Val Arg Thr Lys Lys Ile

100 105 110

Phe Val Gly Gly Leu Pro Pro Ala Leu Thr Ser Asp Glu Phe Arg Ala

115 120 125

Tyr Phe Glu Thr Tyr Gly Pro Val Ser Asp Ala Val Ile Met Ile Asp

130 135 140

Gln Thr Thr Gln Arg Pro Arg Gly Phe Gly Phe Val Ser Phe Asp Ser

145 150 155 160

Glu Asp Ser Val Asp Leu Val Leu His Lys Thr Phe His Asp Leu Asn

165 170 175

Gly Lys Gln Val Glu Val Lys Arg Ala Leu Pro Lys Asp Ala Asn Pro

180 185 190

Gly Ile Ala Ser Gly Gly Gly Arg Gly Ser Gly Gly Ala Gly Gly Phe

195 200 205

Pro Gly Tyr Gly Gly Ser Gly Gly Ser Gly Tyr Glu Gly Arg Val Asp

210 215 220

Ser Asn Arg Tyr Met Gln Pro Gln Asn Thr Gly Ser Gly Tyr Pro Pro

225 230 235 240

Tyr Gly Gly Ser Gly Tyr Gly Thr Gly Tyr Gly Tyr Gly Ser Asn Gly

245 250 255

Val Gly Tyr Gly Gly Phe Gly Gly Tyr Gly Asn Pro Ala Gly Ala Pro

260 265 270

Tyr Gly Asn Pro Ser Val Pro Gly Ala Gly Phe Gly Ser Gly Pro Arg

275 280 285

Ser Ser Trp Gly Ala Gln Ala Pro Ser Gly Tyr Gly Asn Val Gly Tyr

290 295 300

Gly Asn Ala Ala Pro Trp Gly Gly Ser Gly Gly Pro Gly Ser Ala Val

305 310 315 320

Met Gly Gln Ala Gly Ala Ser Ala Gly Tyr Gly Ser Gln Gly Tyr Gly

325 330 335

Tyr Gly Gly Asn Asp Ser Ser Tyr Gly Thr Pro Ser Ala Tyr Gly Ala

340 345 350

Val Gly Gly Arg Ser Gly Asn Met Pro Asn Asn His Gly Gly Gly Gly

355 360 365

Tyr Ala Asp Ala Leu Asp Gly Ser Gly Gly Tyr Gly Asn His Gln Gly

370 375 380

Asn Asn Gly Gln Ala Gly Tyr Gly Gly Gly Tyr Gly Ser Gly Arg Gln

385 390 395 400

Ala Gln Gln Gln

<210>26

<211>2731

<212>DNA

＜213〉Arabidopis thaliana

<400>26

tgagcattgc ttatttgctt ccatccattt ttgttccttt taattcgatt tggattgcag 60

aaaaagaaaa gaaaagaaaa gactaaaaat ttggacgata agcagaaaag agagaggagg 120

gcctctcgcc ctcttattaa aaccttgcct tctccaaatc tgaagatttc tcaatcctaa 180

aatctttttt ttttcctctt tctccgtttc tttattttcg gtattacaca catacataga 240

ttctctgtct tctgggtttt tcattccttc cttcctccaa gcttacacct ttattgatca 300

tttgtgtttt tttttgtttc tgcaggaatc caagatcgtg ggtcgatcgg tttttacaca 360

atccgatcac gacccatctg ctctttttca tcctattttg cttcccttga ggtgtttcta 420

tcgattccat tctccttctc acttagatcg atatagaatc tggaaccaaa aacaaacctt 480

tttttgtttg tttggcagaa atggaaatgg aatcatgtaa gctcttcatc ggtggtatat 540

cttgggaaac cagtgaagat cgtcttcgtg actattttca cagttttggt gaggttttag 600

aggctgttat tatgaaggat cgtgccactg gccgtgctcg tggctttggt ttcgttgtct 660

ttgctgatcc taatgttgct gaaagagtcg tcttgcttaa acatatcatt gatggtaaaa 720

ttgtaagttt cctcctgcta tataccaaca tacattgctt ccaatttcaa caatcttcct 780

gcttacttgc ttcattttga ggttgctgct tctcaaagca aagcaaagct actcactttt 840

attccttcct gttttagtta gtagactcta ttgtttacaa tcagctttgc cgctctgata 900

aatgcatatc tttgtcagaa gttgttcatt tcacactcac aaataaaaat gtaaaacttg 960

gatcgtttca tatcctcatg tgaaagaaag tggttcacaa tgaatgaaaa actgctttct 1020

ttgagttgtg tcgtgtgttg attttctcca tgatatacag gttgaggcaa agaaggctgt 1080

tccaagagat gatcacgtag tatttaataa aagtaacagc agccttcagg gatcacctgg 1140

cccatcaaac tccaagaaga tctttgtggg aggtttggca tcatccgtga cagaggctga 1200

gttcaaaaag tattttgctc agtttgggat gatcactgat gttgtggtga tgtatgacca 1260

cagaacccag cggcctagag gctttgggtt catttcatat gactctgagg aagctgttga 1320

caaagtactg cagaagacat tccacgaact caatggtaag atggtggagg tcaaactggc 1380

tgttcctaag gatatggctc tcaacacaat gcggaaccaa atgaatgtaa atagctttgg 1440

cactagtaga atcagttcat tactgaatga gtacacccag ggattcagcc cgagtccaat 1500

ctctggttat ggagtgaaac ctgaagttag gtacagtcca gcagtaggta ataggggagg 1560

attctcaccg tttggacatg gatacggaat cgagctgaat tttgagccaa accagactca 1620

gaactacggt tctggttcca gtggaggctt tggacgaccc tttagccctg gatatgctgc 1680

gagtctcggc aggttcggta gccaaatgga gtcgggagga gctagtgttg ggaacggttc 1740

tgtcctaaat gcagcaccaa agaaccattt atggggaaat ggtggtctag gttacatgtc 1800

aaactctccg atatcaagaa gcagcttcag tggaaactct ggaatgtctt cactaggcag 1860

cattggtgac aactggggaa cagttgcacg tgcacgcagt agctaccacg gtgagagagg 1920

aggtgtagga ttagaagcaa tgagaggagt tcatgttggt ggttacagca gcggctcaag 1980

catcttggag gcagactctc tgtacagcga ctcgatgtgg ctttcgctgc ctgcaaaggc 2040

agaggaagga ttgggaatgg gaccattgga cttcatgtct agaggaccag ctggatacat 2100

caacaggcaa ccaaacggag gtatgaataa tgaatgaatg aacgcctttt ttctatccga 2160

gaattcaagc atttgtagaa aatctgatga tatcatatga aaatggtgtt gttgcaggaa 2220

ttgcagctta gagaagtgac aaatctatac catggagatc agatgattgc agaagagagt 2280

ttttagaaga ggaaaaaagt ttattaaaaa aaaaaaaatt attggtacca aaaagcttaa 2340

agcttttatt tactttttac tattttgatt tgttgttata gctttctttt cacccttttt 2400

tctaatttgg ggttttgttt cttttgtttt tatcgttaaa gaaaaaagat gtaaacttga 2460

gtgatataaa aagagacaaa gaaacaatga agtgtatttt gttcttgtct ttctctctct 2520

tttatcatct aaatccatat attgacaaat tcaaacatga aaacgaatta aaaaaagagc 2580

aatttgccta gaatgtaggc aacgtagtgt gaggacgacg tgtggcaaac atgtggatga 2640

tgataagcca caggacaaag aaagcaatcc ctcatccatc gcaataatat ccattaatgt 2700

gaagtggacc aaaagagaga gaagcgagtg t 2731

<210>27

<211>431

<212>PRT

＜213〉Arabidopis thaliana

<400>27

Met Glu Met Glu Ser Cys Lys Leu Phe Ile Gly Gly Ile Ser Trp Glu

1 5 10 15

Thr Ser Glu Asp Arg Leu Arg Asp Tyr Phe His Ser Phe Gly Glu Val

20 25 30

Leu Glu Ala Val Ile Met Lys Asp Arg Ala Thr Gly Arg Ala Arg Gly

35 40 45

Phe Gly Phe Val Val Phe Ala Asp Pro Asn Val Ala Glu Arg Val Val

50 55 60

Leu Leu Lys His Ile Ile Asp Gly Lys Ile Val Glu Ala Lys Lys Ala

65 70 75 80

Val Pro Arg Asp Asp His Val Val Phe Asn Lys Ser Asn Ser Ser Leu

85 90 95

Gln Gly Ser Pro Gly Pro Ser Asn Ser Lys Lys Ile Phe Val Gly Gly

100 105 110

Leu Ala Ser Ser Val Thr Glu Ala Glu Phe Lys Lys Tyr Phe Ala Gln

115 120 125

Phe Gly Met Ile Thr Asp Val Val Val Met Tyr Asp His Arg Thr Gln

130 135 140

Arg Pro Arg Gly Phe Gly Phe Ile Ser Tyr Asp Ser Glu Glu Ala Val

145 150 155 160

Asp Lys Val Leu Gln Lys Thr Phe His Glu Leu Asn Gly Lys Met Val

165 170 175

Glu Val Lys Leu Ala Val Pro Lys Asp Met Ala Leu Asn Thr Met Arg

180 185 190

Asn Gln Met Asn Val Asn Ser Phe Gly Thr Ser Arg Ile Ser Ser Leu

195 200 205

Leu Asn Glu Tyr Thr Gln Gly Phe Ser Pro Ser Pro Ile Ser Gly Tyr

210 215 220

Gly Val Lys Pro Glu Val Arg Tyr Ser Pro Ala Val Gly Asn Arg Gly

225 230 235 240

Gly Phe Ser Pro Phe Gly His Gly Tyr Gly Ile Glu Leu Asn Phe Glu

245 250 255

Pro Asn Gln Thr Gln Asn Tyr Gly Ser Gly Ser Ser Gly Gly Phe Gly

260 265 270

Arg Pro Phe Ser Pro Gly Tyr Ala Ala Ser Leu Gly Arg Phe Gly Ser

275 280 285

Gln Met Glu Ser Gly Gly Ala Ser Val Gly Asn Gly Ser Val Leu Asn

290 295 300

Ala Ala Pro Lys Asn His Leu Trp Gly Asn Gly Gly Leu Gly Tyr Met

305 310 315 320

Ser Asn Ser Pro Ile Ser Arg Ser Ser Phe Ser Gly Asn Ser Gly Met

325 330 335

Ser Ser Leu Gly Ser Ile Gly Asp Asn Trp Gly Thr Val Ala Arg Ala

340 345 350

Arg Ser Ser Tyr His Gly Glu Arg Gly Gly Val Gly Leu Glu Ala Met

355 360 365

Arg Gly Val His Val Gly Gly Tyr Ser Ser Gly Ser Ser Ile Leu Glu

370 375 380

Ala Asp Ser Leu Tyr Ser Asp Ser Met Trp Leu Ser Leu Pro Ala Lys

385 390 395 400

Ala Glu Glu Gly Leu Gly Met Gly Pro Leu Asp Phe Met Ser Arg Gly

405 410 415

Pro Ala Gly Tyr Ile Asn Arg Gln Pro Asn Gly Gly Ile Ala Ala

420 425 430

<210>28

<211>1395

<212>DNA

＜213〉rice

<400>28

atggagtcgg atcaggggaa gctgttcatc ggcggcatct cgtgggagac caccgaggag 60

aagctccgcg accacttcgc cgcctacggc gacgtctccc aggccgccgt catgcgcgac 120

aagctcaccg gccgcccccg cggcttcggc ttcgtcgtct tctccgaccc ttcctccgtc 180

gacgccgccc tcgtcgaccc ccacaccctc gacggccgca cggttgatgt gaagcgggcg 240

ctctcgcggg aggagcagca ggccgcgaag gcggcgaacc ctagcgcggg ggggaggcac 300

gcctccggtg ggggcggtgg tgggggaggc gccggtggtg gtggtggtgg cggcggtggt 360

gacgccggcg gtgcgcggac gaagaagatc ttcgtcggcg ggctgccctc caacctgacg 420

gaggacgagt tccggcagta cttccagacc tacggggtcg tcaccgacgt cgtcgtcatg 480

tacgaccaga acacgcagcg gccgaggggg ttcgggttca tcaccttcga cgcggaggac 540

gccgttgacc gcgtgctgca caagaccttc catgacctga gcgggaagat ggtggaggtg 600

aagcgcgccc tgcccaggga ggccaaccct ggctccggca gtggtggccg ttccatggga 660

ggcggcggtg ggggttacca gagtaacaat gggccgaact ccaattctgg gggctatgat 720

agcagaggtg acgctagcag gtatggtcag gcgcagcagg gtagtggtgg ttatcccggt 780

tatggtgctg gaggatatgg tgctggtacg gttggttatg gatatgggca tgctaaccct 840

ggaactgcgt atgggaatta tggggctgga ggatttggag gtgttcctgc tgggtatggt 900

gggcattatg gcaatccaaa tgcgcctggt tcaggttacc agggtggtcc tccaggagca 960

aacagaggac catggggtgg tcaagctccg tctggttatg gcactgggag ttatggtggc 1020

aatgcaggct atgctgcttg gaacaactct tctgctggag gtaatgcacc cactagtcag 1080

gccgctggtg caggcacagg ctatgggagc cagggctatg gatatggtgg atatggagga 1140

gatgcatcgt atggtaatca tggtggatat gggggttatg gaggaagggg agatggtgct 1200

ggcaatccag ctgctggcgg tggatctggg tatggtgctg gctatggaag cgggaatggc 1260

ggttctggtt atccaaatgc ttgggctgat ccttcacaag gtggagggtt tggggcttca 1320

gtcaatggag tgtctgaagg ccaatcaaat tatggcagtg gttatggtgg tgtgcaacct 1380

agggttgctc agtaa 1395

<210>29

<211>464

<212>PRT

＜213〉rice

<400>29

Met Glu Ser Asp Gln Gly Lys Leu Phe Ile Gly Gly Ile Ser Trp Glu

1 5 10 15

Thr Thr Glu Glu Lys Leu Arg Asp His Phe Ala Ala Tyr Gly Asp Val

20 25 30

Ser Gln Ala Ala Val Met Arg Asp Lys Leu Thr Gly Arg Pro Arg Gly

35 40 45

Phe Gly Phe Val Val Phe Ser Asp Pro Ser Ser Val Asp Ala Ala Leu

50 55 60

Val Asp Pro His Thr Leu Asp Gly Arg Thr Val Asp Val Lys Arg Ala

65 70 75 80

Leu Ser Arg Glu Glu Gln Gln Ala Ala Lys Ala Ala Asn Pro Ser Ala

85 90 95

Gly Gly Arg His Ala Ser Gly Gly Gly Gly Gly Gly Gly Gly Ala Gly

100 105 110

Gly Gly Gly Gly Gly Gly Gly Gly Asp Ala Gly Gly Ala Arg Thr Lys

115 120 125

Lys Ile Phe Val Gly Gly Leu Pro Ser Asn Leu Thr Glu Asp Glu Phe

130 135 140

Arg Gln Tyr Phe Gln Thr Tyr Gly Val Val Thr Asp Val Val Val Met

145 150 155 160

Tyr Asp Gln Asn Thr Gln Arg Pro Arg Gly Phe Gly Phe Ile Thr Phe

165 170 175

Asp Ala Glu Asp Ala Val Asp Arg Val Leu His Lys Thr Phe His Asp

180 185 190

Leu Ser Gly Lys Met Val Glu Val Lys Arg Ala Leu Pro Arg Glu Ala

195 200 205

Asn Pro Gly Ser Gly Ser Gly Gly Arg Ser Met Gly Gly Gly Gly Gly

210 215 220

Gly Tyr Gln Ser Asn Asn Gly Pro Asn Ser Asn Ser Gly Gly Tyr Asp

225 230 235 240

Ser Arg Gly Asp Ala Ser Arg Tyr Gly Gln Ala Gln Gln Gly Ser Gly

245 250 255

Gly Tyr Pro Gly Tyr Gly Ala Gly Gly Tyr Gly Ala Gly Thr Val Gly

260 265 270

Tyr Gly Tyr Gly His Ala Asn Pro Gly Thr Ala Tyr Gly Asn Tyr Gly

275 280 285

Ala Gly Gly Phe Gly Gly Val Pro Ala Gly Tyr Gly Gly His Tyr Gly

290 295 300

Asn Pro Asn Ala Pro Gly Ser Gly Tyr Gln Gly Gly Pro Pro Gly Ala

305 310 315 320

Asn Arg Gly Pro Trp Gly Gly Gln Ala Pro Ser Gly Tyr Gly Thr Gly

325 330 335

Ser Tyr Gly Gly Asn Ala Gly Tyr Ala Ala Trp Asn Asn Ser Ser Ala

340 345 350

Gly Gly Asn Ala Pro Thr Ser Gln Ala Ala Gly Ala Gly Thr Gly Tyr

355 360 365

Gly Ser Gln Gly Tyr Gly Tyr Gly Gly Tyr Gly Gly Asp Ala Ser Tyr

370 375 380

Gly Asn His Gly Gly Tyr Gly Gly Tyr Gly Gly Arg Gly Asp Gly Ala

385 390 395 400

Gly Asn Pro Ala Ala Gly Gly Gly Ser Gly Tyr Gly Ala Gly Tyr Gly

405 410 415

Ser Gly Asn Gly Gly Ser Gly Tyr Pro Asn Ala Trp Ala Asp Pro Ser

420 425 430

Gln Gly Gly Gly Phe Gly Ala Ser Val Asn Gly Val Ser Glu Gly Gln

435 440 445

Ser Asn Tyr Gly Ser Gly Tyr Gly Gly Val Gln Pro Arg Val Ala Gln

450 455 460

<210>30

<211>2469

<212>DNA

＜213〉rice

<400>30

ggtccattat ttataccatt tccgcgtccc cccaccctcc tcccccgctt tcccaatcga 60

ggcgagcacc gcaattgcag ggttccggag gccgaataaa aaagtttggc ctctccccgc 120

aaaaaagtaa aaaacccaaa acaaccatcc accagcgcat cgcggcaccg cgagcgagcg 180

agcggaggga gggaggtgga gagcaaaagt tcgataaaag gagaggagga gacgaagcgt 240

cgaagcccaa gtaacatccc cccaacctcc gcctcctcct cctccccctc ctcccatgcc 300

cgcatcgaga tcttagccgc gccggagatc gagagggagg agcggcgacg cgggcgcccc 360

cgatccctcc tcctcgccgc cgccgccgcc ggcggcgccg gagcagcagc agccgacgac 420

gacgacgacc gccgcagcag ccgatcgggg gaggagggga ggggaggacg cgatggaggc 480

ggactccggg aagctcttcg tcggcggcat ctcgtgggag acggacgagg accgcctccg 540

cgagtacttc agccggttcg gggaggtcac cgaggccgtc atcatgcggg accgcaacac 600

cggccgcgcc cgtgggttcg gcttcgtggt cttcaccgac gcaggcgtcg ccgagcgggt 660

caccatggat aagcacatga tcgacgggcg catggtggaa gcgaagaaag ctgttcccag 720

ggacgaccag agcatcacca gcaagaacaa tggcagcagc atagggtcac ctggaccagg 780

ccgtactaga aagatctttg ttggaggctt ggcctctaat gttactgagg ttgaatttag 840

aaggtatttt gagcaatttg gtgtgattac ggatgtggtt gtcatgtacg accacaacac 900

gcagaggcct aggggctttg gattcatcac ctatgactca gaagatgcgg tggacaaggc 960

actgcacaag aacttccatg agctgaatgg taagatggtt gaggtcaaga gagctgttcc 1020

aaaggagcaa tcacctggac ctgctgcacg ttcacctgcg ggagggcaga actatgctat 1080

gagcagggtc catagcttct tgaatggttt caaccagggt tataacccaa accctattgg 1140

aggttatggc atgagggttg atggaaggta tggtctgctt acaggcgcac ggaatggatt 1200

ctcttcattt ggccctggtt atggaatggg catgaattct gaatctggga tgaatgcgaa 1260

ttttggcgcc aattctagtt ttgtcaataa ctccaatggg cggcagatag gttcattcta 1320

caatggtagt tcaaacagat taggtagtcc tattggttat gttggtctta atgatgattc 1380

aggatcacta ttgagttcaa tgtcaaggaa tgtttggggt aatgaaaatc tgaactaccc 1440

aaacaacccc acaaacatga gttcttttgc accatctgga actggaggtc aaatgggtat 1500

taccagtgac ggtattaatt ggggagggcc tactcctggc catggaatgg gcaacatttc 1560

aagccttggg ctggctaacc ttggccgtgg agctggagac agttttggct tgccttctgg 1620

cagctatgga aggagcaatg caactggtac cattggtgaa cccttctctg caccacccaa 1680

tgcatatgaa gtgaacaatg cagatacata tggcagcagc tccatttatg gagactcaac 1740

ttggaggttc acgtcatctg agattgatat gcctcctttt ggtaatgacc ttggaaatgt 1800

tgatccagat atcaaatcaa acataccagc aagttacatg ggcaactata ctgttaataa 1860

taatcagaca agcagaggta tcacttccta gcgagagtac tattatattc atatatgact 1920

tgggatagat gaaagaagca ttatatcagg tattcaggtg catgactatg aattggtgat 1980

atcaggttaa tatacgggtt agttaattgt ttctagctaa ccagaggtgt ggtttatgga 2040

caccaccatg ctagaggagc gaatacaaac gttttgtgaa ggtttcagat tttagtttaa 2100

ttcctacatg tattaggtct tggtttttga atgagatgtg cagtggtgat tgcggcacat 2160

acttagagtg ttccaacata agctggaatc ctgtcatatg gacaaacttg tataccaaag 2220

gaatgcttta ttatcttgcc catttatggc tacattagct cgcttgtttt cattcccttt 2280

ttaaccaatt ccatttgtat actagagatc tgcttgactt actagtgaaa ctattcgggg 2340

acgccgatcc tatctttgca gttggctccc agaaataaag ccaccaaaag tgcatactta 2400

tttgttctac cttgatttgc catatgtata tgcttctgtt cgttttaaaa tagaactttg 2460

ggtttgatt 2469

<210>31

<211>472

<212>PRT

＜213〉rice

<400>31

Met Glu Ala Asp Ser Gly Lys Leu Phe Val Gly Gly Ile Ser Trp Glu

1 5 10 15

Thr Asp Glu Asp Arg Leu Arg Glu Tyr Phe Ser Arg Phe Gly Glu Val

20 25 30

Thr Glu Ala Val Ile Met Arg Asp Arg Asn Thr Gly Arg Ala Arg Gly

35 40 45

Phe Gly Phe Val Val Phe Thr Asp Ala Gly Val Ala Glu Arg Val Thr

50 55 60

Met Asp Lys His Met Ile Asp Gly Arg Met Val Glu Ala Lys Lys Ala

65 70 75 80

Val Pro Arg Asp Asp Gln Ser Ile Thr Ser Lys Asn Asn Gly Ser Ser

85 90 95

Ile Gly Ser Pro Gly Pro Gly Arg Thr Arg Lys Ile Phe Val Gly Gly

100 105 110

Leu Ala Ser Asn Val Thr Glu Val Glu Phe Arg Arg Tyr Phe Glu Gln

115 120 125

Phe Gly Val Ile Thr Asp Val Val Val Met Tyr Asp His Asn Thr Gln

130 135 140

Arg Pro Arg Gly Phe Gly Phe Ile Thr Tyr Asp Ser Glu Asp Ala Val

145 150 155 160

Asp Lys Ala Leu His Lys Asn Phe His Glu Leu Asn Gly Lys Met Val

165 170 175

Glu Val Lys Arg Ala Val Pro Lys Glu Gln Ser Pro Gly Pro Ala Ala

180 185 190

Arg Ser Pro Ala Gly Gly Gln Asn Tyr Ala Met Ser Arg Val His Ser

195 200 205

Phe Leu Asn Gly Phe Asn Gln Gly Tyr Asn Pro Asn Pro Ile Gly Gly

210 215 220

Tyr Gly Met Arg Val Asp Gly Arg Tyr Gly Leu Leu Thr Gly Ala Arg

225 230 235 240

Asn Gly Phe Ser Ser Phe Gly Pro Gly Tyr Gly Met Gly Met Asn Ser

245 250 255

Glu Ser Gly Met Asn Ala Asn Phe Gly Ala Asn Ser Ser Phe Val Asn

260 265 270

Asn Ser Asn Gly Arg Gln Ile Gly Ser Phe Tyr Asn Gly Ser Ser Asn

275 280 285

Arg Leu Gly Ser Pro Ile Gly Tyr Val Gly Leu Asn Asp Asp Ser Gly

290 295 300

Ser Leu Leu Ser Ser Met Ser Arg Asn Val Trp Gly Asn Glu Asn Leu

305 310 315 320

Asn Tyr Pro Asn Asn Pro Thr Asn Met Ser Ser Phe Ala Pro Ser Gly

325 330 335

Thr Gly Gly Gln Met Gly Ile Thr Ser Asp Gly Ile Asn Trp Gly Gly

340 345 350

Pro Thr Pro Gly His Gly Met Gly Asn Ile Ser Ser Leu Gly Leu Ala

355 360 365

Asn Leu Gly Arg Gly Ala Gly Asp Ser Phe Gly Leu Pro Ser Gly Ser

370 375 380

Tyr Gly Arg Ser Asn Ala Thr Gly Thr Ile Gly Glu Pro Phe Ser Ala

385 390 395 400

Pro Pro Asn Ala Tyr Glu Val Asn Asn Ala Asp Thr Tyr Gly Ser Ser

405 410 415

Ser Ile Tyr Gly Asp Ser Thr Trp Arg Phe Thr Ser Ser Glu Ile Asp

420 425 430

Met Pro Pro Phe Gly Asn Asp Leu Gly Asn Val Asp Pro Asp Ile Lys

435 440 445

Ser Asn Ile Pro Ala Ser Tyr Met Gly Asn Tyr Thr Val Asn Asn Asn

450 455 460

Gln Thr Ser Arg Gly Ile Thr Ser

465 470

<210>32

<211>2315

<212>DNA

＜213〉rice

<400>32

ttggagatag aatagagaga gacacacaaa cacctacaac accaacaaca acaagagaaa 60

gagagaaaga agagaaggaa aggagaggaa gaagaggtgg tggtggtggt ggtggtggtg 120

tgtggcctcc ttcccctccc tcctctcgcg aggttgccat gcctccccca agatcgatcc 180

aacccgatca tcaatcgggg cggggaagga ggaggagggg atggaggcgg acgccgggaa 240

gctgttcatc ggcggcatct cgtgggacac caacgaggac cgcctccgcg agtacttcga 300

caagtacggc gaggtggtgg aggccgtcat catgcgcgac cgcgccaccg gccgcgcccg 360

gggattcggc ttcatcgtct tcgctgaccc tgccgtcgcc gagcgggtca ttatggagaa 420

gcacatgatc gatggccgca tggtggaggc gaagaaagct gttcccaggg acgatcagca 480

cgctcttagc aagagcggcg ggagcgctca tggatcgccg gggcccagcc gcaccaagaa 540

gatattcgtt ggggggctag cgtccaccgt gacggaggcg gacttcagga agtactttga 600

gcagttcggg acgatcaccg atgtcgtggt gatgtatgat cacaacacgc agcgtcccag 660

aggttttggg ttcattacgt acgattcgga ggatgctgtg gacaaggcat tgttcaagac 720

cttccatgaa ctgaacggta agatggttga ggtcaagcgc gcggttccta aggaactatc 780

acctgggcct agcatgcgtt ctcctgtcgg tggattcaac tatgccgtga acagagccaa 840

taacttcctc aatggataca cccagggtta taatccgagc ccagtcggtg gctatggaat 900

gaggatggat gcaaggtttg ggcttctatc gggtggccgt agtagttatc cttcttttgg 960

tggtggttat ggagtcggta tgaattttga tccagggatg aaccctgcta ttgggggaag 1020

ctcaagcttc aacaacagtc tccagtatgg aaggcagctt aatccatact acagtggaaa 1080

ttctggtaga tacaatagca atgttagcta tggtggagtc aatgacagta ctgggtcagt 1140

gttcaactcg ctggctcgta atttatgggg taattcaggt cttagttact cttccaactc 1200

tgcaagctct aattccttca tgtcatctgc caatgggggc cttggtggaa ttgggaacaa 1260

caatgtgaat tggggaaacc ctcctgtgcc tgcacaaggt gctaatgctg gcccaggcta 1320

tggcagtggg aacttcggtt atggatccag tgaaaccaac tttggtctcg gtaccaatgc 1380

ttatggaagg aatgctggat ctggtgttgt taatacattc aatcaatcaa ccaatgggta 1440

tggaaggaac tttggagatt catcaggagg aggtggcggt ggtggcggtg gctccatcta 1500

tggagacaca acttggagat ccggatcttc tgagcttgat ggaaccagcc catttggcta 1560

tgggcttggg aatgcagctt cagatgttac agcaaagaac tcagcaggtt acatggggca 1620

ttaacaaata gagcaatgtc gccgcctagg aatctttttc acatacaaca tttgtcaaaa 1680

taggttgagg agagaaccac aggtgcatca ggtgcaaatt ttgaacctca catgatttac 1740

agaaatgggt tagttaatag agctaaccac cagggatttg gtcaatgaga tcagatatat 1800

atcctcagag aaccatttaa acgtatttcc attttatgta aggtttgaga ttgtggtttc 1860

ggatttctac agcgagttta ggttttggca accttgtgtt ttttcttggt tgagatgtga 1920

agtaagattg cgggatatat atatctgaag agtgttcagt tgtacggcgg cgctgccccc 1980

atataggccc ccctttttgg gtttttgttc ttatagtaga aactgctcta gcgttttgca 2040

aattgtgtgc tagctgttgt tatcaggatg ataatttttt tccccttctt ggtttttatc 2100

ttactgaagt gtatgtacca gagatcttgc tggtctgtgt ttttcctagt ggaacttttg 2160

agggatgccc cttctgggtc tcaaagaata ataatgctac attatattct aattcatttt 2220

gaggctttct aaggctatat attatttgta tgtaccctgc tggaacatct gtacattctg 2280

atgctctttg caatttgcct ttgtgctgct tttgc 2315

<210>33

<211>467

<212>PRT

＜213〉rice

<400>33

Met Glu Ala Asp Ala Gly Lys Leu Phe Ile Gly Gly Ile Ser Trp Asp

1 5 10 15

Thr Asn Glu Asp Arg Leu Arg Glu Tyr Phe Asp Lys Tyr Gly Glu Val

20 25 30

Val Glu Ala Val Ile Met Arg Asp Arg Ala Thr Gly Arg Ala Arg Gly

35 40 45

Phe Gly Phe Ile Val Phe Ala Asp Pro Ala Val Ala Glu Arg Val Ile

50 55 60

Met Glu Lys His Met Ile Asp Gly Arg Met Val Glu Ala Lys Lys Ala

65 70 75 80

Val Pro Arg Asp Asp Gln His Ala Leu Ser Lys Ser Gly Gly Ser Ala

85 90 95

His Gly Ser Pro Gly Pro Ser Arg Thr Lys Lys Ile Phe Val Gly Gly

100 105 110

Leu Ala Ser Thr Val Thr Glu Ala Asp Phe Arg Lys Tyr Phe Glu Gln

115 120 125

Phe Gly Thr Ile Thr Asp Val Val Val Met Tyr Asp His Asn Thr Gln

130 135 140

Arg Pro Arg Gly Phe Gly Phe Ile Thr Tyr Asp Ser Glu Asp Ala Val

145 150 155 160

Asp Lys Ala Leu Phe Lys Thr Phe His Glu Leu Asn Gly Lys Met Val

165 170 175

Glu Val Lys Arg Ala Val Pro Lys Glu Leu Ser Pro Gly Pro Ser Met

180 185 190

Arg Ser Pro Val Gly Gly Phe Asn Tyr Ala Val Asn Arg Ala Asn Asn

195 200 205

Phe Leu Asn Gly Tyr Thr Gln Gly Tyr Asn Pro Ser Pro Val Gly Gly

210 215 220

Tyr Gly Met Arg Met Asp Ala Arg Phe Gly Leu Leu Ser Gly Gly Arg

225 230 235 240

Ser Ser Tyr Pro Ser Phe Gly Gly Gly Tyr Gly Val Gly Met Asn Phe

245 250 255

Asp Pro Gly Met Asn Pro Ala Ile Gly Gly Ser Ser Ser Phe Asn Asn

260 265 270

Ser Leu Gln Tyr Gly Arg Gln Leu Asn Pro Tyr Tyr Ser Gly Asn Ser

275 280 285

Gly Arg Tyr Asn Ser Asn Val Ser Tyr Gly Gly Val Asn Asp Ser Thr

290 295 300

Gly Ser Val Phe Asn Ser Leu Ala Arg Asn Leu Trp Gly Asn Ser Gly

305 310 315 320

Leu Ser Tyr Ser Ser Asn Ser Ala Ser Ser Asn Ser Phe Met Ser Ser

325 330 335

Ala Asn Gly Gly Leu Gly Gly Ile Gly Asn Asn Asn Val Asn Trp Gly

340 345 350

Asn Pro Pro Val Pro Ala Gln Gly Ala Asn Ala Gly Pro Gly Tyr Gly

355 360 365

Ser Gly Asn Phe Gly Tyr Gly Ser Ser Glu Thr Asn Phe Gly Leu Gly

370 375 380

Thr Asn Ala Tyr Gly Arg Asn Ala Gly Ser Gly Val Val Asn Thr Phe

385 390 395 400

Asn Gln Ser Thr Asn Gly Tyr Gly Arg Asn Phe Gly Asp Ser Ser Gly

405 410 415

Gly Gly Gly Gly Gly Gly Gly Gly Ser Ile Tyr Gly Asp Thr Thr Trp

420 425 430

Arg Ser Gly Ser Ser Glu Leu Asp Gly Thr Ser Pro Phe Gly Tyr Gly

435 440 445

Leu Gly Asn Ala Ala Ser Asp Val Thr Ala Lys Asn Ser Ala Gly Tyr

450 455 460

Met Gly His

465

<210>34

<211>1146

<212>DNA

＜213〉common wheat (Triticum aestivum)

<400>34

aaaaagcagg tgggaccggc ccggaattct cgggatatcg tcgacccacg cgtccgcgca 60

cccgagcgcg agagaatccg aggagaggag cggcgcaagg aggcggtgat ggagtcggat 120

cagggcaagc tcttcatcgg cggcatctcc tgggagacga cggaggagaa gctgcaggag 180

cacttctcca acttcggcga ggtctcccag gccgccgtca tgcgcgacaa gctcactggc 240

cgcccgcggg gcttcggctt cgtagtctac gccgaccccg ccgccgtcga cgccgccctc 300

caggagcccc acaccctcga cggccgcacg gtcgatgtga agcgggcgct ctcgcgggag 360

gagcagcagg ctaccaaggc ggtgaaccct agcgcaggaa ggaacgctgg aggtggtggc 420

ggcggcggcg gcggcggcgg cgatgccggt ggtgctagga caaagaagat ttttgtgggc 480

ggactgccct ccagtctgac agatgaggag ttccggcagt acttccagac cttcggggct 540

gtcaccgatg ttgtggtgat gtatgaccag acaacacagc gtccccgggg cttcggcttc 600

attacctttg actcggagga tgcggttgac cgtgtgctgc acaaaacctt ccacgatctt 660

ggagggaaga tggtagaggt gaagcgtgct ctgccccgag aggcgaatcc tggctctggc 720

ggcggcggcc gttccatggg aggtgggggg tttcatagta acaatggacc ccactccaat 780

gctagcagct atgatggcag aggcgatgct agcagatatg ggcaggcgca gcaaggcatg 840

ggtggctacc caggttatgg tgctggagct tatggcagtg ctccaactgg gtttggatat 900

gggccaccca atccgggaac tacttatgga aatattgggt ctgcagggtt aggagctttt 960

ccttggtgcg tatgcggggg gcttatgggc aacccaggtg gctgcgggtt tcgggttacc 1020

cgggggggcc cctccggggc cctaaataag ggaccctggg ggcagccaaa cctccgccct 1080

ggtttatggc acctgggggc tttatcctgg gcacgtgcgg ggctattggg tgcgtggaaa 1140

taaccc 1146

<210>35

<211>344

<212>PRT

＜213〉common wheat

<400>35

Met Glu Ser Asp Gln Gly Lys Leu Phe Ile Gly Gly Ile Ser Trp Glu

1 5 10 15

Thr Thr Glu Glu Lys Leu Gln Glu His Phe Ser Asn Phe Gly Glu Val

20 25 30

Ser Gln Ala Ala Val Met Arg Asp Lys Leu Thr Gly Arg Pro Arg Gly

35 40 45

Phe Gly Phe Val Val Tyr Ala Asp Pro Ala Ala Val Asp Ala Ala Leu

50 55 60

Gln Glu Pro His Thr Leu Asp Gly Arg Thr Val Asp Val Lys Arg Ala

65 70 75 80

Leu Ser Arg Glu Glu Gln Gln Ala Thr Lys Ala Val Asn Pro Ser Ala

85 90 95

Gly Arg Asn Ala Gly Gly Gly Gly Gly Gly Gly Gly Gly Gly Gly Asp

100 105 110

Ala Gly Gly Ala Arg Thr Lys Lys Ile Phe Val Gly Gly Leu Pro Ser

115 120 125

Ser Leu Thr Asp Glu Glu Phe Arg Gln Tyr Phe Gln Thr Phe Gly Ala

130 135 140

Val Thr Asp Val Val Val Met Tyr Asp Gln Thr Thr Gln Arg Pro Arg

145 150 155 160

Gly Phe Gly Phe Ile Thr Phe Asp Ser Glu Asp Ala Val Asp Arg Val

165 170 175

Leu His Lys Thr Phe His Asp Leu Gly Gly Lys Met Val Glu Val Lys

180 185 190

Arg Ala Leu Pro Arg Glu Ala Asn Pro Gly Ser Gly Gly Gly Gly Arg

195 200 205

Ser Met Gly Gly Gly Gly Phe His Ser Asn Asn Gly Pro His Ser Asn

210 215 220

Ala Ser Ser Tyr Asp Gly Arg Gly Asp Ala Ser Arg Tyr Gly Gln Ala

225 230 235 240

Gln Gln Gly Met Gly Gly Tyr Pro Gly Tyr Gly Ala Gly Ala Tyr Gly

245 250 255

Ser Ala Pro Thr Gly Phe Gly Tyr Gly Pro Pro Asn Pro Gly Thr Thr

260 265 270

Tyr Gly Asn Ile Gly Ser Ala Gly Leu Gly Ala Phe Pro Trp Cys Val

275 280 285

Cys Gly Gly Leu Met Gly Asn Pro Gly Gly Cys Gly Phe Arg Val Thr

290 295 300

Arg Gly Gly Pro Ser Gly Ala Leu Asn Lys Gly Pro Trp Gly Gln Pro

305 310 315 320

Asn Leu Arg Pro Gly Leu Trp His Leu Gly Ala Leu Ser Trp Ala Arg

325 330 335

Ala Gly Leu Leu Gly Ala Trp Lys

340

<210>36

<211>800

<212>DNA

＜213〉sugarcane (Saccharum officinarum)

<220>

<221>misc_feature

<222>(8)..(8)

＜223〉n is a, c, g or t

<220>

<221>misc_feature

<222>(381)..(381)

＜223〉n is a, c, g or t

<220>

<221>misc_feature

<222>(425)..(425)

＜223〉n is a, c, g or t

<220>

<221>misc_feature

<222>(428)..(428)

＜223〉n is a, c, g or t

<220>

<221>misc_feature

<222>(481)..(481)

＜223〉n is a, c, g or t

<220>

<221>misc_feature

<222>(514)..(514)

＜223〉n is a, c, g or t

<400>36

agaattcncg gttcgaccta cgcgtccgcc cggaatcccc aattccgctc tcttcctctc 60

tccctctctc ccccaccgca gcatcaggcg agcgcgaggc ggaggtggag gagagatgga 120

gttggaccag ggcaagctct tcatcggcgg catctcctgg gagacgacgg aggagaagct 180

gagcgagcac ttctccgcct acggcgaggt tacgcaggcc gccgtcatgc gggacaagat 240

caccggccgc ccccgtggct tcgggttcgt cgtcttcgcc gaccccgccg tcgtcgaccg 300

agcgctgcag gacccccaca ccctcgacgg ccgcacggtc gatgtgaagc gggcactctc 360

gcgggaggag cagcaggcct ncaaggccgc gaaccctagc ggtgggagga acactggcgg 420

tggangangc ggcgggtggc ggggcggcga tgcaagtggt gctcggaccc aggaagatct 480

ntggggggcc ggcttgcctt ctactctgac tganggatgg gtttcggcag tactttccgg 540

accttcggag gggtcactga tggttggtgg ccatggttga accggaacaa gcaattgccc 600

gcgttggttt tggaatcaat acttttgaac tttaagattc cggtgaaccg ctgctggcca 660

agaactttca tgacctggtg ggaagatggt ttaaggtgaa ccagcattgc gcccttgagg 720

cgaaccctgg gggttctgga acgggccgtt ctgggggaaa tgggggcttt ctagcaacca 780

tggccttacc cccgttttgg 800

<210>37

<211>1243

<212>DNA

＜213〉rice

<400>37

aaaaccaccg agggacctga tctgcaccgg ttttgatagt tgagggaccc gttgtgtctg 60

gttttccgat cgagggacga aaatcggatt cggtgtaaag ttaagggacc tcagatgaac 120

ttattccgga gcatgattgg gaagggagga cataaggccc atgtcgcatg tgtttggacg 180

gtccagatct ccagatcact cagcaggatc ggccgcgttc gcgtagcacc cgcggtttga 240

ttcggcttcc cgcaaggcgg cggccggtgg ccgtgccgcc gtagcttccg ccggaagcga 300

gcacgccgcc gccgccgacc cggctctgcg tttgcaccgc cttgcacgcg atacatcggg 360

atagatagct actactctct ccgtttcaca atgtaaatca ttctactatt ttccacattc 420

atattgatgt taatgaatat agacatatat atctatttag attcattaac atcaatatga 480

atgtaggaaa tgctagaatg acttacattg tgaattgtga aatggacgaa gtacctacga 540

tggatggatg caggatcatg aaagaattaa tgcaagatcg tatctgccgc atgcaaaatc 600

ttactaattg cgctgcatat atgcatgaca gcctgcatgc gggcgtgtaa gcgtgttcat 660

ccattaggaa gtaaccttgt cattacttat accagtacta catactatat agtattgatt 720

tcatgagcaa atctacaaaa ctggaaagca ataagaaata cgggactgga aaagactcaa 780

cattaatcac caaatatttc gccttctcca gcagaatata tatctctcca tcttgatcac 840

tgtacacact gacagtgtac gcataaacgc agcagccagc ttaactgtcg tctcaccgtc 900

gcacactggc cttccatctc aggctagctt tctcagccac ccatcgtaca tgtcaactcg 960

gcgcgcgcac aggcacaaat tacgtacaaa acgcatgacc aaatcaaaac caccggagaa 1020

gaatcgctcc cgcgcgcggc ggcgacgcgc acgtacgaac gcacgcacgc acgcccaacc 1080

ccacgacacg atcgcgcgcg acgccggcga caccggccgt ccacccgcgc cctcacctcg 1140

ccgactataa atacgtaggc atctgcttga tcttgtcatc catctcacca ccaaaaaaaa 1200

aaggaaaaaa aaacaaaaca caccaagcca aataaaagcg aca 1243

<210>38

<211>154

<212>PRT

＜213〉sugarcane

<220>

<221>misc_feature

<222>(89)..(89)

＜223〉Xaa can be any naturally occurring amino acid

<220>

<221>misc_feature

<222>(104)..(105)

＜223〉Xaa can be any naturally occurring amino acid

<220>

<221>misc_feature

<222>(133)..(133)

＜223〉Xaa can be any naturally occurring amino acid

<400>38

Met Glu Leu Asp Gln Gly Lys Leu Phe Ile Gly Gly Ile Ser Trp Glu

1 5 10 15

Thr Thr Glu Glu Lys Leu Ser Glu His Phe Ser Ala Tyr Gly Glu Val

20 25 30

Thr Gln Ala Ala Val Met Arg Asp Lys Ile Thr Gly Arg Pro Arg Gly

35 40 45

Phe Gly Phe Val Val Phe Ala Asp Pro Ala Val Val Asp Arg Ala Leu

50 55 60

Gln Asp Pro His Thr Leu Asp Gly Arg Thr Val Asp Val Lys Arg Ala

65 70 75 80

Leu Ser Arg Glu Glu Gln Gln Ala Xaa Lys Ala Ala Asn Pro Ser Gly

85 90 95

Gly Arg Asn Thr Gly Gly Gly Xaa Xaa Gly Gly Trp Arg Gly Gly Asp

100 105 110

Ala Ser Gly Ala Arg Thr Gln Glu Asp Leu Trp Gly Ala Gly Leu Pro

115 120 125

Ser Thr Leu Thr Xaa Gly Trp Val Ser Ala Val Leu Ser Gly Pro Ser

130 135 140

Glu Gly Ser Leu Met Val Gly Gly His Gly

145 150

<210>39

<211>59

<212>DNA

＜213〉artificial sequence

<220>

＜223〉primer: prm00405

<400>39

ggggacaagt ttgtacaaaa aagcaggctt cacaatggat tatgatcggt acaagttat 59

<210>40

<211>54

<212>DNA

＜213〉artificial sequence

<220>

＜223〉primer: prm00406

<400>40

ggggaccact ttgtacaaga aagctgggtt taaaagagtc caaagaattt cact 54

Claims (36)

1. the method for improvement plant growth characteristics, it comprises the activity that increases rna binding protein in the plant or its homologue, wherein said rna binding protein or its homologue are following arbitrary:

(i) has RNA in conjunction with active polypeptide and comprise 2 or 3 RNA identification motif RRM and have the motif of at least 75% sequence identity and/or have at least 50% sequence identity motif with motif II:RFDPFTGEPYKFDP with motif I:PYEAAVVALPVVVKERLVRILRLGIATRYD; Or

(ii) RBP1 polypeptide or its homologue, it has (a) RNA in conjunction with activity; (b) two RRM structural domains; (c) following two motifs: (i) KIFVGGL and (ii) RPRGFGF allow three aminoacid replacement and conservative arbitrarily change at the most in the motif; (d) amino acid with SEQ ID NO:15 representative has at least 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95% sequence identity; With

(iii) randomly select to have the plant of improvement growth characteristics.

2. according to the process of claim 1 wherein that the activity of described increase is to introduce genetic modification by locus conjugated protein at coding RNA or its homologue preferably to realize.

3. according to the method for claim 2, wherein said genetic modification is the arbitrary realization in activating by site-directed mutagenesis, homologous recombination, TILLING or T-DNA.

4. improve the method for plant growth characteristics, it is included in the plant and introduces and the conjugated protein coding nucleic acid of expressed rna or its functional variant, and the rna binding protein of wherein said coding or its homologue are following arbitrary:

(i) has RNA in conjunction with active polypeptide and comprise 2 or 3 RNA identification motif RRM and have the motif of at least 75% sequence identity and/or the motif that has at least 50% sequence identity with motif II:RFDPFTGEPYKFDP with motif I:PYEAAVVALPVVVKERLVRILRLGIATRYD; Or

(ii) RBP1 polypeptide or its homologue, it has (a) RNA in conjunction with activity; (b) two RRM structural domains; (c) following two motifs: (i) KIFVGGL and (ii) RPRGFGF allow three aminoacid replacement and conservative arbitrarily change at the most in the motif; (d) amino acid with SEQ ID NO:15 representative has at least 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95% sequence identity.

5. according to the method for claim 4, wherein said variant be the rna binding protein coding nucleic acid part or can with the sequence of rna binding protein coding nucleic acid hybridization.

6. according to the method for claim 4 or 5, wherein said rna binding protein coding nucleic acid or its functional variant are crossed in plant and are expressed.

7. according to each method in the claim 4 to 6, wherein the described rna binding protein coding nucleic acid of claim 4 (i) or its functional variant are plant origins, and preferably from dicotyledons, further preferably from tobacco section, more preferably nucleic acid is from tobacco.

8. according to each method in the claim 4 to 6, wherein the (ii) described rna binding protein coding nucleic acid of claim 4 or its functional variant are plant origins, preferably from dicotyledons, further preferably from Cruciferae, more preferably nucleic acid is from Arabidopis thaliana.

9. according to each method in the claim 4 to 7, wherein said rna binding protein coding nucleic acid or its functional variant are connected in the promotor of seed preference effectively.

10. according to the method for claim 9, wherein said promotor is the prolamine promotor, as the prolamine promotor from rice.

11. according to each method in claim 4 to 6 and the claim 8, wherein said rbp1 nucleic acid or its functional variant are connected in effectively and can express the promotor of described nucleic acid in preference ground in bud.

12. according to the method for claim 11, wherein said promotor has and the similar express spectra of β expansion protein promoter.

13. according to each method in the claim 1 to 12, the plant growth characteristics of wherein said improvement is the productive rate that increases with respect to corresponding wild type plant.

14. according to each method in the claim 1 to 13, the plant growth characteristics of wherein said improvement is the phytomass that increases.

15. according to the method for claim 13, the productive rate of wherein said increase is the seed productive rate that increases.

16. according to the method for claim 15, the seed productive rate of wherein said increase is selected from the following seed biomass of appointing one or more (i) to increase; (ii) (full) seed number of Zeng Jiaing; The (iii) seed size of Zeng Jiaing; The (iv) seed volume of Zeng Jiaing; (the v) harvest index of Zeng Jiaing; (the vi) thousand seed weight of Zeng Jiaing (TKW).

17. according to each method in the claim 1 to 16, the plant growth characteristics of wherein said improvement is the growth velocity that increases.

18. can be according to the plant of each method acquisition in the claim 1 to 17.

19. construct comprises:

(i) rna binding protein coding nucleic acid or its variant, this rna binding protein have RNA in conjunction with active and comprise 2 or 3 RNA identification motif RRM and have the motif of at least 75% sequence identity and/or the motif that has at least 50% sequence identity with motif II:RFDPFTGEPYKFDP with motif I:PYEAAVVALPVVVKERLVRILRLGIATRYD;

(ii) can drive one or more control sequences of the nucleotide sequence expression of (i); With optional

(iii) transcription termination sequence.

20. construct comprises:

(i) rbp1 coding nucleic acid or its variant, the RBP1 of wherein said coding has: (a) RNA is in conjunction with activity; (b) two RRM structural domains; (c) following two motifs: (i) KIFVGGL and (ii) RPRGFGF allow three aminoacid replacement and conservative arbitrarily change at the most in the motif; (d) amino acid with SEQ ID NO:15 representative has at least 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95% sequence identity;

(ii) can drive one or more control sequences of the nucleotide sequence expression of (i); With optional

(iii) transcription termination sequence.

21. according to the construct of claim 19, wherein said control sequence is to drive expression promoter in seed tissue.

22. according to the construct of claim 21, wherein said promotor is the prolamine promotor, as the prolamine promotor from rice.

23. according to the construct of claim 20, wherein said promotor can drive the expression in bud.

24. according to the construct of claim 23, wherein said promotor has similar express spectra to β expansion protein promoter.

25. use according to each construct plant transformed in the claim 19 to 24.

26. produce the method for the transgenic plant with improvement growth characteristics, this method comprises:

(i) introduce rna binding protein coding nucleic acid or its variant in plant, the rna binding protein of its coding has RNA in conjunction with active and comprise 2 or 3 RNA identification motif RRM and have the motif that the motif of at least 75% sequence identity and/or motif and motif II:RFDPFTGEPYKFDP have at least 50% sequence identity with motif I:PYEAAVVALPVVVKERLVRILRLGIATRYD; With

(ii) culturing plants cell under the condition that promotes plant-growth and growth.

27. produce the method for the transgenic plant with improvement growth characteristics, this method comprises:

(i) introduce rbp1 coding nucleic acid or its variant in plant, the RBP1 of wherein said coding has: (a) RNA is in conjunction with activity; (b) two RRM structural domains; (c) following two motifs: (i) KIFVGGL and (ii) RPRGFGF allow three aminoacid replacement and conservative arbitrarily change at the most in the motif; (d) amino acid with SEQ ID NO:15 representative has at least 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95% sequence identity; With

(ii) culturing plants cell under the condition that promotes plant-growth and growth.

28. have the transgenic plant of improvement growth characteristics, it is by rna binding protein coding nucleic acid or its variant are introduced described plant generation.

29. have the transgenic plant of improvement growth characteristics, it is by rbp1 nucleic acid or its variant are introduced described plant generation.

30. according to claim 18,25 or 28 and 29 transgenic plant, wherein said plant is a monocotyledons, as sugarcane, perhaps wherein said plant is a cereal, as rice, corn, wheat, barley, grain, rye, oat or Chinese sorghum.

31. the part gathered in the crops according to each plant in the claim 18,25 or 28 to 30.

32. according to the part gathered in the crops of claim 31, wherein said gather in the crops the part be seed.

33.RNA conjugated protein coding nucleic acid/gene or its variant or rna binding protein or its homologue particularly improve the especially purposes in the seed productive rate of productive rate at the improvement plant growth characteristics.

34.rbp1 or its variant or RBP1 polypeptide or its homologue particularly improve the especially purposes in the seed productive rate of productive rate at the improvement plant growth characteristics.

35. according to the purposes of claim 33 or 34, wherein said seed productive rate comprises following one or more: (full) seed number of increase, the seed weight of increase, the harvest index of increase and the TKW of increase.

36.RNA conjugated protein or its variant or rbp1 or its variant are as the purposes of molecule marker.

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