CN1184231C - Wheat TaDREB, its code gene and method for culturing adverse-resistant plant - Google Patents
Wheat TaDREB, its code gene and method for culturing adverse-resistant plant Download PDFInfo
- Publication number
- CN1184231C CN1184231C CNB011365692A CN01136569A CN1184231C CN 1184231 C CN1184231 C CN 1184231C CN B011365692 A CNB011365692 A CN B011365692A CN 01136569 A CN01136569 A CN 01136569A CN 1184231 C CN1184231 C CN 1184231C
- Authority
- CN
- China
- Prior art keywords
- plant
- ala
- tadreb
- ser
- glu
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Expired - Fee Related
Links
- 241000196324 Embryophyta Species 0.000 title claims abstract description 52
- 108090000623 proteins and genes Proteins 0.000 title claims abstract description 31
- 241000209140 Triticum Species 0.000 title claims abstract description 26
- 235000021307 Triticum Nutrition 0.000 title claims abstract description 26
- 238000000034 method Methods 0.000 title claims abstract description 16
- 230000002411 adverse Effects 0.000 title abstract description 9
- 238000012258 culturing Methods 0.000 title 1
- 108091023040 Transcription factor Proteins 0.000 claims abstract description 22
- 102000040945 Transcription factor Human genes 0.000 claims abstract description 22
- 239000013604 expression vector Substances 0.000 claims description 17
- 241000589158 Agrobacterium Species 0.000 claims description 12
- 238000006243 chemical reaction Methods 0.000 claims description 5
- 125000003275 alpha amino acid group Chemical group 0.000 claims description 4
- 230000001404 mediated effect Effects 0.000 claims description 4
- 241000209510 Liliopsida Species 0.000 claims description 3
- 241000193830 Bacillus <bacterium> Species 0.000 claims description 2
- 241001233957 eudicotyledons Species 0.000 claims description 2
- 239000002773 nucleotide Substances 0.000 claims description 2
- 125000003729 nucleotide group Chemical group 0.000 claims description 2
- 239000002245 particle Substances 0.000 claims description 2
- 239000013598 vector Substances 0.000 claims description 2
- 238000004113 cell culture Methods 0.000 claims 1
- 239000002299 complementary DNA Substances 0.000 description 9
- 102220023256 rs387907547 Human genes 0.000 description 7
- 241000282326 Felis catus Species 0.000 description 5
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 description 5
- 108020004999 messenger RNA Proteins 0.000 description 5
- 108090000790 Enzymes Proteins 0.000 description 4
- 102000004190 Enzymes Human genes 0.000 description 4
- 150000003839 salts Chemical class 0.000 description 4
- 108020004414 DNA Proteins 0.000 description 3
- 108091034057 RNA (poly(A)) Proteins 0.000 description 3
- 108091081024 Start codon Proteins 0.000 description 3
- 244000098338 Triticum aestivum Species 0.000 description 3
- 235000001014 amino acid Nutrition 0.000 description 3
- 150000001413 amino acids Chemical class 0.000 description 3
- 239000003623 enhancer Substances 0.000 description 3
- 239000012634 fragment Substances 0.000 description 3
- 239000013612 plasmid Substances 0.000 description 3
- 239000011780 sodium chloride Substances 0.000 description 3
- 108091032973 (ribonucleotides)n+m Proteins 0.000 description 2
- IJGRMHOSHXDMSA-UHFFFAOYSA-N Atomic nitrogen Chemical compound N#N IJGRMHOSHXDMSA-UHFFFAOYSA-N 0.000 description 2
- 241000894006 Bacteria Species 0.000 description 2
- 241000701489 Cauliflower mosaic virus Species 0.000 description 2
- HEDRZPFGACZZDS-UHFFFAOYSA-N Chloroform Chemical compound ClC(Cl)Cl HEDRZPFGACZZDS-UHFFFAOYSA-N 0.000 description 2
- ZRALSGWEFCBTJO-UHFFFAOYSA-N Guanidine Chemical compound NC(N)=N ZRALSGWEFCBTJO-UHFFFAOYSA-N 0.000 description 2
- 240000007594 Oryza sativa Species 0.000 description 2
- 235000007164 Oryza sativa Nutrition 0.000 description 2
- 238000004458 analytical method Methods 0.000 description 2
- 102220369446 c.1274G>A Human genes 0.000 description 2
- 238000010276 construction Methods 0.000 description 2
- 238000013016 damping Methods 0.000 description 2
- 239000012530 fluid Substances 0.000 description 2
- 230000001939 inductive effect Effects 0.000 description 2
- 230000026731 phosphorylation Effects 0.000 description 2
- 238000006366 phosphorylation reaction Methods 0.000 description 2
- 238000012545 processing Methods 0.000 description 2
- 238000013138 pruning Methods 0.000 description 2
- 238000003757 reverse transcription PCR Methods 0.000 description 2
- 235000009566 rice Nutrition 0.000 description 2
- 239000000523 sample Substances 0.000 description 2
- 239000000126 substance Substances 0.000 description 2
- 230000009466 transformation Effects 0.000 description 2
- 238000013519 translation Methods 0.000 description 2
- 229910052902 vermiculite Inorganic materials 0.000 description 2
- 235000019354 vermiculite Nutrition 0.000 description 2
- 239000010455 vermiculite Substances 0.000 description 2
- 238000005406 washing Methods 0.000 description 2
- FRXSZNDVFUDTIR-UHFFFAOYSA-N 6-methoxy-1,2,3,4-tetrahydroquinoline Chemical compound N1CCCC2=CC(OC)=CC=C21 FRXSZNDVFUDTIR-UHFFFAOYSA-N 0.000 description 1
- 240000008067 Cucumis sativus Species 0.000 description 1
- 235000010799 Cucumis sativus var sativus Nutrition 0.000 description 1
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 description 1
- CEAZRRDELHUEMR-URQXQFDESA-N Gentamicin Chemical compound O1[C@H](C(C)NC)CC[C@@H](N)[C@H]1O[C@H]1[C@H](O)[C@@H](O[C@@H]2[C@@H]([C@@H](NC)[C@@](C)(O)CO2)O)[C@H](N)C[C@@H]1N CEAZRRDELHUEMR-URQXQFDESA-N 0.000 description 1
- 229930182566 Gentamicin Natural products 0.000 description 1
- 102000003960 Ligases Human genes 0.000 description 1
- 108090000364 Ligases Proteins 0.000 description 1
- 108060001084 Luciferase Proteins 0.000 description 1
- 239000005089 Luciferase Substances 0.000 description 1
- 235000007688 Lycopersicon esculentum Nutrition 0.000 description 1
- 240000004658 Medicago sativa Species 0.000 description 1
- 235000010624 Medicago sativa Nutrition 0.000 description 1
- CHJJGSNFBQVOTG-UHFFFAOYSA-N N-methyl-guanidine Natural products CNC(N)=N CHJJGSNFBQVOTG-UHFFFAOYSA-N 0.000 description 1
- 101150005851 NOS gene Proteins 0.000 description 1
- 108700026244 Open Reading Frames Proteins 0.000 description 1
- ISWSIDIOOBJBQZ-UHFFFAOYSA-N Phenol Chemical compound OC1=CC=CC=C1 ISWSIDIOOBJBQZ-UHFFFAOYSA-N 0.000 description 1
- 108700001094 Plant Genes Proteins 0.000 description 1
- 241000220317 Rosa Species 0.000 description 1
- 241000124033 Salix Species 0.000 description 1
- 240000003768 Solanum lycopersicum Species 0.000 description 1
- 238000002105 Southern blotting Methods 0.000 description 1
- NINIDFKCEFEMDL-UHFFFAOYSA-N Sulfur Chemical compound [S] NINIDFKCEFEMDL-UHFFFAOYSA-N 0.000 description 1
- 239000005864 Sulphur Substances 0.000 description 1
- 230000010632 Transcription Factor Activity Effects 0.000 description 1
- 240000008042 Zea mays Species 0.000 description 1
- 235000005824 Zea mays ssp. parviglumis Nutrition 0.000 description 1
- 235000002017 Zea mays subsp mays Nutrition 0.000 description 1
- 239000002253 acid Substances 0.000 description 1
- 239000003513 alkali Substances 0.000 description 1
- 239000003242 anti bacterial agent Substances 0.000 description 1
- 229940088710 antibiotic agent Drugs 0.000 description 1
- 239000002585 base Substances 0.000 description 1
- 238000010523 cascade reaction Methods 0.000 description 1
- 239000003153 chemical reaction reagent Substances 0.000 description 1
- 150000001875 compounds Chemical class 0.000 description 1
- 238000007796 conventional method Methods 0.000 description 1
- 235000005822 corn Nutrition 0.000 description 1
- 229960000935 dehydrated alcohol Drugs 0.000 description 1
- 238000011161 development Methods 0.000 description 1
- 230000018109 developmental process Effects 0.000 description 1
- SWSQBOPZIKWTGO-UHFFFAOYSA-N dimethylaminoamidine Natural products CN(C)C(N)=N SWSQBOPZIKWTGO-UHFFFAOYSA-N 0.000 description 1
- 230000000694 effects Effects 0.000 description 1
- 238000004520 electroporation Methods 0.000 description 1
- 238000005516 engineering process Methods 0.000 description 1
- 230000007613 environmental effect Effects 0.000 description 1
- 230000006353 environmental stress Effects 0.000 description 1
- 239000013613 expression plasmid Substances 0.000 description 1
- 238000013467 fragmentation Methods 0.000 description 1
- 238000006062 fragmentation reaction Methods 0.000 description 1
- 230000002068 genetic effect Effects 0.000 description 1
- 229960002518 gentamicin Drugs 0.000 description 1
- 239000004009 herbicide Substances 0.000 description 1
- YQYJSBFKSSDGFO-FWAVGLHBSA-N hygromycin A Chemical compound O[C@H]1[C@H](O)[C@H](C(=O)C)O[C@@H]1Oc1ccc(\C=C(/C)C(=O)N[C@@H]2[C@@H]([C@H]3OCO[C@H]3[C@@H](O)[C@@H]2O)O)cc1O YQYJSBFKSSDGFO-FWAVGLHBSA-N 0.000 description 1
- 238000003780 insertion Methods 0.000 description 1
- 230000037431 insertion Effects 0.000 description 1
- 230000003834 intracellular effect Effects 0.000 description 1
- JEIPFZHSYJVQDO-UHFFFAOYSA-N iron(III) oxide Inorganic materials O=[Fe]O[Fe]=O JEIPFZHSYJVQDO-UHFFFAOYSA-N 0.000 description 1
- 239000007788 liquid Substances 0.000 description 1
- 238000004519 manufacturing process Methods 0.000 description 1
- 239000003550 marker Substances 0.000 description 1
- 238000000520 microinjection Methods 0.000 description 1
- 239000000203 mixture Substances 0.000 description 1
- 229910052757 nitrogen Inorganic materials 0.000 description 1
- 102000039446 nucleic acids Human genes 0.000 description 1
- 108020004707 nucleic acids Proteins 0.000 description 1
- 150000007523 nucleic acids Chemical class 0.000 description 1
- 230000031787 nutrient reservoir activity Effects 0.000 description 1
- 230000001151 other effect Effects 0.000 description 1
- 238000004806 packaging method and process Methods 0.000 description 1
- 229920001184 polypeptide Polymers 0.000 description 1
- 239000002244 precipitate Substances 0.000 description 1
- 239000002243 precursor Substances 0.000 description 1
- 238000000327 preparative centrifugation Methods 0.000 description 1
- 102000004196 processed proteins & peptides Human genes 0.000 description 1
- 108090000765 processed proteins & peptides Proteins 0.000 description 1
- 239000000047 product Substances 0.000 description 1
- 235000018102 proteins Nutrition 0.000 description 1
- 102000004169 proteins and genes Human genes 0.000 description 1
- 238000000746 purification Methods 0.000 description 1
- 238000011160 research Methods 0.000 description 1
- 238000010839 reverse transcription Methods 0.000 description 1
- JQXXHWHPUNPDRT-WLSIYKJHSA-N rifampicin Chemical compound O([C@](C1=O)(C)O/C=C/[C@@H]([C@H]([C@@H](OC(C)=O)[C@H](C)[C@H](O)[C@H](C)[C@@H](O)[C@@H](C)\C=C\C=C(C)/C(=O)NC=2C(O)=C3C([O-])=C4C)C)OC)C4=C1C3=C(O)C=2\C=N\N1CC[NH+](C)CC1 JQXXHWHPUNPDRT-WLSIYKJHSA-N 0.000 description 1
- 229960001225 rifampicin Drugs 0.000 description 1
- 238000005096 rolling process Methods 0.000 description 1
- 102220023258 rs387907548 Human genes 0.000 description 1
- 238000012216 screening Methods 0.000 description 1
- 238000012882 sequential analysis Methods 0.000 description 1
- 230000011869 shoot development Effects 0.000 description 1
- YZHUMGUJCQRKBT-UHFFFAOYSA-M sodium chlorate Chemical compound [Na+].[O-]Cl(=O)=O YZHUMGUJCQRKBT-UHFFFAOYSA-M 0.000 description 1
- 239000007787 solid Substances 0.000 description 1
- 239000006228 supernatant Substances 0.000 description 1
- 239000000725 suspension Substances 0.000 description 1
- 230000005026 transcription initiation Effects 0.000 description 1
- 230000002103 transcriptional effect Effects 0.000 description 1
- 230000009261 transgenic effect Effects 0.000 description 1
- 230000014621 translational initiation Effects 0.000 description 1
- 239000005418 vegetable material Substances 0.000 description 1
- 235000013311 vegetables Nutrition 0.000 description 1
- 239000013603 viral vector Substances 0.000 description 1
Images
Landscapes
- Breeding Of Plants And Reproduction By Means Of Culturing (AREA)
Abstract
The present invention provides new wheat TaDREB genes which comprise DREB transcription factors of an AP2/EREBP structural domain. The present invention also provides a method for cultivating adverse resisting plant by using the genes.
Description
Technical field
The present invention relates to a kind of AP
2/ EREBP class transcription factor DREB specifically describes, and the present invention relates to a kind of AP that derives from wheat (Triticum aestivum)
2/ EREBP class transcription factor DREB, called after TaDREB.The invention still further relates to the gene of this class transcription factor of coding, contain the carrier of this genoid and utilize this genoid to cultivate the method for plant with adverse resistance.
Technical background
The variation of physical chemical factor in the environment, for example: arid, saline and alkaline and damage to plants caused by sudden drop in temperature and make the extensive underproduction of farm crop.Therefore, cultivating the resistance of reverse crop is one of major objective of plant husbandry.Higher plant cell can have number of ways to experience the variation of physico-chemical parameter in the external environment, thereby the signal that born of the same parents are outer becomes intracellular signal, phosphorylation cascade reaction by series passes to transcription factor with signal, transcription factor remakes and is used for functional gene, thereby the expression of gene that the startup adverse circumstance is replied improves the resistance of reverse of plant.In plant, AP
2Class in the/EREBP class DREB transcription factor family can be accepted the environment-stress signal and start the adverse circumstance response gene.Wheat (Triticum aestivum) is an important crops.Cultivate and resist cold, salt tolerant, drought-enduring crop new variety are one of vital tasks in the Wheat Production.The contriver has cloned DREB transcription factor, called after TaDREB from wheat.This expression of gene is subjected to arid, salt, cold inducing.Make up the expression vector that is applicable to single, double cotyledon plant respectively, transformed unifacial leaf model plant paddy rice.The tangible resistance of reverse of transfer-gen plant tool.
Summary of the invention
An object of the present invention is to provide a kind of TaDREB transcription factor with stronger anti-reverse effect, this transcription factor derives from wheat (Triticum aestivum), and its aminoacid sequence is shown in Fig. 1 and SEQ ID NO:2.Sequence shown in Fig. 1 has been represented DREB transcription factor family in the wheat.Should be pointed out that transcription factor of the present invention is carried out one or more amino acid whose replacements, insertion and/or lack resulting functional analogue also reaching purpose of the present invention.Thereby the present invention also comprises with the aminoacid sequence shown in the SEQ ID NO:2 having at least 90% homology, preferably has at least 95% homology, but has the functional analogue of wheat DREB transcription factor activity simultaneously.
Another object of the present invention provides the gene of a kind of encoding transcription factor TaDREB, has represented a kind of DREB transcription factor family in the wheat at the gene shown in Fig. 1 and the SEQ ID NO:1, and they all contain conservative AP
2/ EREBP structural domain sequence.But be not limited to listed sequence.
A further object of the present invention provides a kind of method of cultivating anti-contrary plant, and this method comprises the gene constructed plant expression vector with transcription factor TaDREB of the present invention; Transform plant tissue with the expression vector that makes up; The plant transformed tissue cultivating is become plant.Described conversion can be undertaken by agriculture bacillus mediated or particle bombardment.
Another purpose of the present invention provides a kind of expression vector (Fig. 2) that contains mentioned TaDREB class transcription factor.Can use any expression vector that can guide foreign gene in plant, to express.These plant expression vectors include but not limited to, the double base agrobacterium vector, for example Bin19 (Bevan, M., 1984, nucleic acids research, 12:8711-8721), the carrier that contains the promotor of DRE element and be used for the monocotyledons micropellet bombardment.Carrier also can comprise 3 ' untranslated zone.3 ' untranslated zone is meant a kind of part of gene, but it contain a kind of comprise the polyadenylic acid signal and any other effect mRNA processing or the dna fragmentation of genetic expression.Polyadenylic acid signal guidance polyadenylic acid joins 3 ' end of mRNA precursor.The example in 3 ' zone comprises that containing the Agrobacterium crown-gall nodule induces (Ti) plasmid gene (for example rouge alkali synthetase (Nos gene)), and the 3 ' non-translational region of transcribing of plant gene (for example big storage protein gene).3 ' untranslated zone of DREB transcription factor of the present invention can be used to express in plant.
Carrier of the present invention also can contain suitable promotor.Can use any strong promoter in the present invention.These promotors include but not limited to cauliflower mosaic virus (CAMV 35S) and Ubiqutin promotor.It can use separately or be used in combination with other plant promoter.
Expression vector of the present invention also can comprise enhanser when needed, no matter be translational enhancer or transcriptional enhancer.These enhanser zones include but not limited to ATG initiator codon and neighboring region.Initiator codon must with the reading frame homophase of encoding sequence, to guarantee the translation of whole sequence.The translation control signal can be multiple different source with initiator codon, can be natural or synthetic.Translation initiation region can be from the transcription initiation zone, or from structure gene.
For the ease of transgenic plant cells or plant being identified and screening, can process employed carrier, comprise adding the alternative mark of plant.Spendable selected marker comprises the enzyme to antibiotics resistance, and microbiotic comprises gentamicin, Totomycin, kantlex etc.Equally, can use the enzyme that produces by the compound of colour-change (for example GUS) or the identification of luminous (for example luciferase) fork-like farm tool used in ancient China, or anti-chemical reagent (for example weedkiller).In addition, also can any selection markers.
Expression vector of the present invention can be by using Ti-plasmids, Ri plasmid, plant viral vector, directly DNA conversion, microinjection, importing such as electroporation vegetable cell.The summary of these technology is consulted Weissbach and Weissbach, molecular biology of plants method (Method for PlantMolecular Biology), Academy Press, New York VIII, pp.421-463 (1988); With Geiserson and Corey, molecular biology of plants (Plant Molecular Biology), second edition (1988).
Can use method plant transformed host of the present invention to comprise monocotyledons and dicotyledons, for example: paddy rice, wheat, corn, cucumber, tomato, willow, turfgrass, lucerne place etc.
Brief Description Of Drawings
Fig. 1 represents the Nucleotide and the deduced amino acid of TaDREB gene;
Fig. 2 represents the structure of TaDREB plant expression vector;
Fig. 3 represents that difference coerces down the TaDREB expression of gene feature that detects through RT-PCR;
Fig. 4 represents the compositional analysis of TaDREB gene in the wheat cdna group.
Embodiment
One, wheat cDNA library construction and transcription factor TaDREB cDNA clone, sequential analysis
Little wheat seeds after 14 days, take out in basin, soak 3-5 hour in 2.0%NaCl solution.The cDNA library construction is undertaken by Sambrook (1987) method.Collect fresh complete stool blade 1g and grind in liquid nitrogen, be suspended from the 4mol/L sulphur hydracid guanidine, mixture adds dehydrated alcohol and precipitates total RNA with acid phenol, chloroform extracting in the supernatant, afterwards, water-soluble.Obtain mRNA through the processing of mRNA purification kit.Get 2ug mRNA and carry out reverse transcription reaction, and then carry out two chains and synthesize.Double-stranded cDNA is connected the back phosphorylation with the EcoR1 joint, (Sephacryl-400) removes unnecessary joint through the preparative centrifugation post, the dephosphorization pExcell that cuts with enzyme then, and carrier (Pharmacia) connects, and gets 5ul connection product adding packaging protein, builds up the cDNA library.
With Arabidopis thaliana DREB cDNA is that probe has screened 4.5 * 10
5The spot that do not increase obtains 3 positive spots, and it is 1.29kb that its skewer is gone into fragment.This fragment comprises the open reading frame of 834bp, the polypeptide that coding is made up of 278 amino acid, and its 5 ' end is 251bp, 3 ' end is 173bp.Wheat TaDREB cDNA complete sequence and aminoacid sequence are seen Fig. 1.
The structure of two .TaDREB cDNA plant expression plasmids and agrobacterium mediation converted Arabidopis thaliana
The structure of TaDREB cDNA plant expression vector carries out according to a conventional method.Binary expression vector pB1121 contains the fragment of 35S promoter and translational enhancer TMV.The plant expression vector that will contain 35S promoter directly transforms Agrobacterium, and environmental Co1-0 (the Arabidopis thaliana biology resource center) plant of Arabidopis thaliana is used for Agrobacterium-mediated Transformation, and agrobacterium strains is EH105 (Inst. of Genetics and Development Biology, CAS).
1. vegetable material is prepared
(1) with the vernalization 72 hours under 4 ℃ of conditions of Arabidopis thaliana Co1-0 seed.With 1/3 * B
5Nutritive medium soaks the plantation basin that vermiculite (vermiculite) is housed, after planting in 23 ℃ of cultivations.
(2) treat plant blossom after, cut off its major branch top, promote the side shoot development.In 4-6 after beta pruning days, set about preparing Agrobacterium-mediated Transformation.
2. Agrobacterium is cultivated
The expression of exogenous gene carrier electric shock of having that (3) will build transforms Agrobacterium, and PCR and enzyme are cut and checked and shake bacterium after errorless.
(4) Agrobacterium is inoculated in the 5ml LB+50 μ g/ml rifampin+50 μ g/ml kantlex substratum, shakes bacterium and spend the night.Rolling bottle to 1 rose in the LB substratum in second day, and 28 ℃ are cultured to OD
600About=0.8.
(5) centrifugal collection thalline transforms damping fluid suspension Agrobacterium with 1 liter.About 300-400ml can transform 2 basin plant.
3. vacuumize conversion
(6) 4-6 after the beta pruning days plant is inverted in contains in the vial that transforms damping fluid, vacuumize, kept 0.05Mpa pressure 5-10 minute.
(7) take out the plantation basin, be sidelong 24 hours.It is upright to plant basin then, cultivates plant to solid by usual method, the results mature seed.
Three, the resistance of reverse of gene plant is identified
Transfer-gen plant and adjoining tree be transplanted to respectively contain 150mM, continued growth in the MS substratum of 200mM and 250mMNaCl, under a plurality of salt concn, transfer-gen plant grows good and adjoining tree wiltings of turning white gradually, all death of back all around.
Four analyses of .TaDREB gene in the wheat cdna group
With TaDREB is probe, carries out Southern Blot and analyze under the rigorous degree of difference, when washing film under 65 ℃, presents many hybrid belts, shows a plurality of copies that exist the TaDREB gene in the wheat cdna group.Banding pattern when washing film under 45 ℃ of low rigorous conditions is consistent with 65 ℃, shows that the TaDREB gene does not have other homology analogue (Fig. 4) in the wheat cdna group.
Five, the expression characteristic of wheat TaDREB in adverse circumstance
With 2.0%NaCl, 20%PEG and 30mM ABA solution and 4 ℃ of subzero treatment wheats 1 day respectively 0,2, were collected blade in 5,24 hours, extracted that RNA makes RT-PCR and Northern analyzes.The result shows that transcribing of TaDREB is subjected to salt in leaf, and arid and ABA coerce and the inducing in various degree of low temperature adverse circumstance (Fig. 3).
Sequence table
<110 〉
1Institute of Genetics, Academia Sinica
Tsing-Hua University
<120〉method of wheat TaDREB, its encoding gene and cultivation plant with adverse resistance
<130>I2001271
<160>2
<170>PatentIn?version?3.1
<210>1
<211>1292
<212>DNA
<213>Triticum?aestivum
<220>
<221>CDS
<222>(252)..(1085)
<223>
<400>1
caaaaccaag?gcggcggcag?cggggcggga?gagcggggag?caccgaccga?caccggccga 60
cagggtgggc?tgcatgcgga?gctgaggcga?ggcgaggcga?ggcggggaga?gatccggcgc 120
gggtgccacc?gccggccggc?cgcgggagat?ctggttggtg?gcgccgcccg?gataagggag 180
aggcggcgag?gggagagcag?ccgggggaga?ccgaggcgag?aggagatctc?tctcgtccct 240
cttctcgctc?c?atg?gag?acc?ggg?ggt?agc?aag?cgg?gaa?gga?gac?tgc?ccc 290
Met?Glu?Thr?Gly?Gly?Ser?Lys?Arg?Glu?Gly?Asp?Cys?Pro
1 5 10
ggg?cag?gaa?agg?aag?aag?aaa?gtg?cgc?agg?aga?agc?act?ggt?cct?gat 338
Gly?Gln?Glu?Arg?Lys?Lys?Lys?Val?Arg?Arg?Arg?Ser?Thr?Gly?Pro?Asp
15 20 25
tcg?gtt?gct?gaa?acc?atc?aag?aag?tgg?aag?gag?gaa?aac?cag?aag?ctc 386
Ser?Val?Ala?Glu?Thr?Ile?Lys?Lys?Trp?Lys?Glu?Glu?Asn?Gln?Lys?Leu
30 35 40 45
cag?caa?gag?aat?gga?tcc?cgg?aaa?gca?ccg?gcc?aag?ggt?tcc?aag?aaa 434
Gln?Gln?Glu?Asn?Gly?Ser?Arg?Lys?Ala?Pro?Ala?Lys?Gly?Ser?Lys?Lys
50 55 60
ggg?tgc?atg?gca?ggg?aaa?gga?ggt?cca?gag?aat?tca?aac?tgc?gct?tac 482
Gly?Cys?Met?Ala?Gly?Lys?Gly?Gly?Pro?Glu?Asn?Ser?Asn?Cys?Ala?Tyr
65 70 75
cgc?ggt?gtg?agg?cag?agg?acg?tgg?ggg?aaa?tgg?gtt?gct?gag?atc?cgt 530
Arg?Gly?Val?Arg?Gln?Arg?Thr?Trp?Gly?Lys?Trp?Val?Ala?Glu?Ile?Arg
80 85 90
gag?ccc?aac?cgt?ggc?aat?cgg?ctg?tgg?ctt?ggt?tca?ttc?cct?acc?gca 578
Glu?Pro?Asn?Arg?Gly?Asn?Arg?Leu?Trp?Leu?Gly?Ser?Phe?Pro?Thr?Ala
95 100 105
gtc?gaa?gct?gca?cgt?gca?tat?gat?gat?gcg?gca?agg?gca?atg?tat?ggc 626
Val?Glu?Ala?Ala?Arg?Ala?Tyr?Asp?Asp?Ala?Ala?Arg?Ala?Met?Tyr?Gly
110 115 120 125
gcc?aaa?gca?cgt?gtc?aac?ttc?tca?gag?cag?tcc?ccg?gat?gcc?aac?tct 674
Ala?Lys?Ala?Arg?Val?Asn?Phe?Ser?Glu?Gln?Ser?Pro?Asp?Ala?Asn?Ser
130 135 140
ggt?tgc?acg?ctg?gca?cct?cca?ttg?ccg?atg?tct?aat?ggg?gca?acc?gct 722
Gly?Cys?Thr?Leu?Ala?Pro?Pro?Leu?Pro?Met?Ser?Asn?Gly?Ala?Thr?Ala
145 150 155
gcg?tca?cat?cct?tct?gat?ggg?aag?gat?gaa?tcg?gag?tct?cct?cct?tct 770
Ala?Ser?His?Pro?Ser?Asp?Gly?Lys?Asp?Glu?Ser?Glu?Ser?Pro?Pro?Ser
160 165 170
ctt?atc?tca?aat?gcg?ccg?aca?gct?gcg?ctg?cat?cgg?tct?gat?gct?aag 818
Leu?Ile?Ser?Asn?Ala?Pro?Thr?Ala?Ala?Leu?His?Arg?Ser?Asp?Ala?Lys
175 180 185
gat?gag?tct?gag?tct?gca?ggg?acc?gtg?gca?cgt?aag?gtg?aaa?aaa?gaa 866
Asp?Glu?Ser?Glu?Ser?Ala?Gly?Thr?Val?Ala?Arg?Lys?Val?Lys?Lys?Glu
190 195 200 205
gtg?agc?aat?gat?ttg?aga?agt?acc?cat?gag?gag?cac?aag?acc?ctg?gaa 914
Val?Ser?Asn?Asp?Leu?Arg?Ser?Thr?His?Glu?Glu?His?Lys?Thr?Leu?Glu
210 215 220
gta?tcc?caa?cca?aaa?ggg?aag?gct?tta?cat?aaa?gca?gcg?aac?gta?agt 962
Val?Ser?Gln?Pro?Lys?Gly?Lys?Ala?Leu?His?Lys?Ala?Ala?Asn?Val?Ser
225 230 235
tat?gat?tac?ttc?aac?gtc?gag?gaa?gtt?ctt?gac?atg?ata?att?gtg?gaa 1010
Tyr?Asp?Tyr?Phe?Asn?Val?Glu?Glu?Val?Leu?Asp?Met?Ile?Ile?Val?Glu
240 245 250
ttg?agt?gct?gat?gta?aaa?atg?gaa?gca?cat?gaa?gag?tac?caa?gat?ggt 1058
Leu?Ser?Ala?Asp?Val?Lys?Met?Glu?Ala?His?Glu?Glu?Tyr?Gln?Asp?Gly
255 260 265
gat?gat?ggg?ttt?agt?ctt?ttc?tca?tat?tagggtttta?gctatgaggg 1105
Asp?Asp?Gly?Phe?Ser?Leu?Phe?Ser?Tyr
270 275
ttgcagtcat?gcggagcaat?agggataact?ttcattctag?ctgctaggaa?atacttcaaa 1165
tctgcaaccc?gaagctttgt?agtcacttat?ggttttcatc?ttactggaga?gaatagcttt 1225
ataccataag?tcaacgggta?caagaagttg?tcctgtgcgt?tgagttcatg?tactatggta 1285
aaagttg 1292
<210>2
<211>278
<212>PRT
<213>Triticum?aestivum
<400>2
Met?Glu?Thr?Gly?Gly?Ser?Lys?Arg?Glu?Gly?Asp?Cys?Pro?Gly?Gln?Glu
1 5 10 15
Arg?Lys?Lys?Lys?Val?Arg?Arg?Arg?Ser?Thr?Gly?Pro?Asp?Ser?Val?Ala
20 25 30
Glu?Thr?Ile?Lys?Lys?Trp?Lys?Glu?Glu?Asn?Gln?Lys?Leu?Gln?Gln?Glu
35 40 45
Asn?Gly?Ser?Arg?Lys?Ala?Pro?Ala?Lys?Gly?Ser?Lys?Lys?Gly?Cys?Met
50 55 60
Ala?Gly?Lys?Gly?Gly?Pro?Glu?Asn?Ser?Asn?Cys?Ala?Tyr?Arg?Gly?Val
65 70 75 80
Arg?Gln?Arg?Thr?Trp?Gly?Lys?Trp?Val?Ala?Glu?Ile?Arg?Glu?Pro?Asn
85 90 95
Arg?Gly?Asn?Arg?Leu?Trp?Leu?Gly?Ser?Phe?Pro?Thr?Ala?Val?Glu?Ala
100 105 110
Ala?Arg?Ala?Tyr?Asp?Asp?Ala?Ala?Arg?Ala?Met?Tyr?Gly?Ala?Lys?Ala
115 120 125
Arg?Val?Asn?Phe?Ser?Glu?Gln?Ser?Pro?Asp?Ala?Asn?Ser?Gly?Cys?Thr
130 135 140
Leu?Ala?Pro?Pro?Leu?Pro?Met?Ser?Asn?Gly?Ala?Thr?Ala?Ala?Ser?His
145 150 155 160
Pro?Ser?Asp?Gly?Lys?Asp?Glu?Ser?Glu?Ser?Pro?Pro?Ser?Leu?Ile?Ser
165 170 175
Asn?Ala?Pro?Thr?Ala?Ala?Leu?His?Arg?Ser?Asp?Ala?Lys?Asp?Glu?Ser
180 185 190
Glu?Ser?Ala?Gly?Thr?Val?Ala?Arg?Lys?Val?Lys?Lys?Glu?Val?Ser?Asn
195 200 205
Asp?Leu?Arg?Ser?Thr?His?Glu?Glu?His?Lys?Thr?Leu?Glu?Val?Ser?Gln
210 215 220
Pro?Lys?Gly?Lys?Ala?Leu?His?Lys?Ala?Ala?Asn?Val?Ser?Tyr?Asp?Tyr
225 230 235 240
Phe?Asn?Val?Glu?Glu?Val?Leu?Asp?Met?Ile?Ile?Val?Glu?Leu?Ser?Ala
245 250 255
Asp?Val?Lys?Met?Glu?Ala?His?Glu?Glu?Tyr?Gln?Asp?Gly?Asp?Asp?Gly
260 265 270
Phe?Ser?Leu?Phe?Ser?Tyr
275
Claims (8)
1. AP who derives from wheat
2/ EREBP class transcription factor TaDREB, it has the aminoacid sequence shown in the SEQ IDNO:2.
2. AP of the described wheat of claim 1 that encodes
2The gene of/EREBP class transcription factor TaDREB.
3. according to the described gene of claim 2, it has the nucleotide sequence shown in the SEQ ID NO:1.
4. plant expression vector that contains the gene of claim 2 or 3 described TaDREB transcription factors.
5. according to the described plant expression vector of claim 4, wherein, described plant expression vector is the double base agrobacterium vector.
6. a method of cultivating the resistance of reverse plant comprises the gene constructed plant expression vector with claim 2 or 3 described TaDREB transcription factors; With the expression vector transformed plant cells that makes up; With the plant transformed cell culture is become plant.
7. in accordance with the method for claim 6, wherein, described conversion is undertaken by agriculture bacillus mediated or particle bombardment.
8. in accordance with the method for claim 6, wherein said plant is monocotyledons or dicotyledons.
Priority Applications (1)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
CNB011365692A CN1184231C (en) | 2001-10-18 | 2001-10-18 | Wheat TaDREB, its code gene and method for culturing adverse-resistant plant |
Applications Claiming Priority (1)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
CNB011365692A CN1184231C (en) | 2001-10-18 | 2001-10-18 | Wheat TaDREB, its code gene and method for culturing adverse-resistant plant |
Publications (2)
Publication Number | Publication Date |
---|---|
CN1412199A CN1412199A (en) | 2003-04-23 |
CN1184231C true CN1184231C (en) | 2005-01-12 |
Family
ID=4673742
Family Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
CNB011365692A Expired - Fee Related CN1184231C (en) | 2001-10-18 | 2001-10-18 | Wheat TaDREB, its code gene and method for culturing adverse-resistant plant |
Country Status (1)
Country | Link |
---|---|
CN (1) | CN1184231C (en) |
Families Citing this family (4)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN100344759C (en) * | 2005-11-08 | 2007-10-24 | 天津师范大学 | cDNA sequence of dehydrolysis responding transcription factor DREB gene of wheal |
CN102234320B (en) * | 2010-04-27 | 2013-07-31 | 中国农业科学院作物科学研究所 | Plant stress-tolerant associated protein TaDREB4B and encoding gene and application thereof |
CN103755793B (en) * | 2014-01-28 | 2016-08-24 | 中国科学院新疆生态与地理研究所 | The most drought-enduring 4 DREB transcription factor of the red moss of tooth rib and encoding gene thereof |
CN104910263B (en) * | 2014-03-12 | 2018-03-13 | 中国农业科学院作物科学研究所 | Plant stress tolerance correlative protein TaPPR and its encoding gene and application |
-
2001
- 2001-10-18 CN CNB011365692A patent/CN1184231C/en not_active Expired - Fee Related
Also Published As
Publication number | Publication date |
---|---|
CN1412199A (en) | 2003-04-23 |
Similar Documents
Publication | Publication Date | Title |
---|---|---|
CN1291021C (en) | Use of boea crassifolia BcBCP1 gene for breeding drought-salt-tolerant plants | |
CN107099544A (en) | The PL LbCpf1 RVR genes of identification specific site and its application in paddy gene target practice | |
CN107142271A (en) | The PL LbCpf1 RR genes with high mutation efficiency and its application in gene targeting | |
CN1223606C (en) | Rice DREB transcription factor and its encoding gene and use | |
CN1184231C (en) | Wheat TaDREB, its code gene and method for culturing adverse-resistant plant | |
CN1824779A (en) | Soyabean thermal excited transcryption factor, its coding gene and application | |
CN1296383C (en) | Plant DREB transcription factor and its coding gene and uses | |
CN1216904C (en) | Soybean ethylene response protein transcription factor and its coded gene and application | |
CN1940072A (en) | Antiglyphosate gene and its use | |
CN1821405A (en) | Xinjiang Saussurea involucrata cold regulation protein and its code gene and use | |
CN1908011A (en) | Plant inverse-resistant zinc finger protein, coding gene and application thereof | |
CN1218959C (en) | Paddy rice ethylene receptor protein, coded gene and use thereof | |
CN1900280A (en) | Rice tiller regulating gene OsTIL1 and its use | |
CN1044385C (en) | Rice genus resisting rice genus leaf stripe virus with transformed behavious and its mfg. method | |
CN1919867A (en) | Soybean Trihelix transcription factor, encode gene and application thereof | |
CN1548453A (en) | Frigostable correlative transcriptive factor of rice and its coding gene and application | |
CN1390937A (en) | Spider silk protein gene designed and synthesized by plant preference codon and its application | |
CN1281754C (en) | Cold resisting application of glycan aldehydic acid enzyme arrestin gene AtPGIP1 of Arabidopsis thalianum | |
CN1255538C (en) | Rice coronal root control gene CRL3 and its use | |
CN1385530A (en) | Method for raising taxol content by gene engineering technology | |
CN112626050B (en) | SpCas9-NRCH mutant for recognizing specific sites in rice gene targeting and application thereof | |
CN1614024A (en) | Transgened-tobacco for degrading organic mercury pollution | |
CN1609209A (en) | Wheat cysteine proteinase gene and its application | |
CN100336904C (en) | Pathogen from rice, chemically inducible promoter and their use | |
CN1778814A (en) | Binding protein of tall fescue drought reacting element, its coding gene and use thereof |
Legal Events
Date | Code | Title | Description |
---|---|---|---|
C10 | Entry into substantive examination | ||
SE01 | Entry into force of request for substantive examination | ||
C06 | Publication | ||
PB01 | Publication | ||
C10 | Entry into substantive examination | ||
SE01 | Entry into force of request for substantive examination | ||
C14 | Grant of patent or utility model | ||
GR01 | Patent grant | ||
C19 | Lapse of patent right due to non-payment of the annual fee | ||
CF01 | Termination of patent right due to non-payment of annual fee |