CN102351950A - Rice drought-tolerance related transcription factor gene OsWTF1, and coding protein and application thereof - Google Patents
Rice drought-tolerance related transcription factor gene OsWTF1, and coding protein and application thereof Download PDFInfo
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Abstract
The invention discloses a rice drought-tolerance related transcription factor gene OsWTF1, and a coding protein and application thereof. The rice drought-tolerance related protein is (a) protein composed of amino acid sequence disclosed as SEQ ID NO:2 in the sequence table; or (b) plant stress-tolerance related protein which is derived from (a) and is one or more amino acid residues of substitution, and/or deletion and/or addition of the amino acid sequence disclosed as SEQ ID NO:2 in the sequence table. The rice drought-tolerance related protein and the coding gene thereof can be used for cultivating drought-tolerant rice.
Description
Technical field
The present invention relates to plant biotechnology field, be specifically related to separating clone, functional verification and the application of the AP2/ERF of a rice transcription factor family superfamily ERF subfamily wax genes involved.
Background technology
Paddy rice is the most important food crop of China, and paddy rice also is first rich and influential family of agriculture water consumption.Owing to the influence of ecotope and climate warming, water resources shortage has seriously restricted the production of China's grain, and has become the major cause of the paddy rice underproduction in recent years.For cultivating siccocolous paddy rice new lines, people attempt being familiar with the reaction mechanism of plant to arid through the biotechnology approach, and utilize molecular biology method to improve the drought resistance of plant.Research shows that plant can secrete wax in stratum corneum surface or stratum corneum, limits the non-gas porosity loss of water of plant, has the effect of water conservation drought resisting.On the other hand; The ERF transcription factor that the further investigation of plant transcription factor is also found to contain 58-59 high conservative amino acid structure territory can participate in regulating the answering that plant is coerced biology and abiotic factor, improves the tolerance effect that plant is coerced arid, sick worm, low temperature adverse circumstance etc.To the more and more serious water shortage problem that China is faced with, can find the ERF class transcription factor of adjusting and controlling rice wax related gene expression, will improve rice yield and have great importance cultivating the rice varieties of novel drought resisting.
Summary of the invention
The purpose of this invention is to provide a kind of wax transcription factor gene and the proteins encoded thereof relevant with paddy drought resistance.The wax transcription factor gene relevant provided by the present invention with paddy drought resistance; Name is called OsWTF1 (with WIN1 homology in the Arabidopis thaliana); This gene is numbered Os02g0202000 in rice genome DB TIGR; The ERF type paddy rice wax metabolism transcription factor of encoding one and containing " AP2 " structural domain, the CDS total length 618bp of this gene, the albumen of coding 205aa.Also has no research report up till now about the OsWTF1 function.Gene according to the invention promptly is the gene that is numbered Os02g0202000 among the rice genome DB TIGR, is one of following nucleotide sequences:
1) dna sequence dna of sequence table SEQ ID No:1;
2) polynucleotide of SEQ ID No:2 protein sequence in the code sequence tabulation;
3) with sequence table in the dna sequence dna that limits of SEQ ID No:1 have the homology 90% or more, and the identical function protein DNA sequence of encoding.
The dna sequence dna of SEQ ID No:1 contains 2 exons by 912 based compositions in the sequence table, be respectively from 5 ' the 154th at end to the 236th bit base and from the 357th at 5 ' end to the 892nd bit base.The nucleotide residue sequence of this sequence encoding SEQ ID No:2.
The proteins encoded OsWTF1 of the gene OsWTF1 relevant with rice drought tolerance; Be protein, or the aminoacid sequence of SEQ ID No:2 is passed through replacement, disappearance or the interpolation of one or several amino-acid residues and has identical active by SEQ ID No:2 deutero-protein with the amino acid residue sequence of SEQ ID No:2 with SEQ ID No:2 amino acid residue sequence.
The protein that SEQ ID No:2 amino acid residue sequence is made up of 205 amino-acid residues in the sequence table.
The expression vector, transgenic cell line and the host bacterium that contain gene OsWTF1 of the present invention all belong to protection scope of the present invention.
Arbitrary segmental primer is to also within protection scope of the present invention among the amplification OsWTF1.
Another object of the present invention provides a kind of method that improves plant drought resistance.
The method of improvement plant drought resistance provided by the present invention is to make up expression vector importing plant tissue or cell with drought resistance genes involved OsWTF1, the drought resistance of improvement plant with said.
The carrier that sets out that is used to make up said plant expression vector comprises double base agrobacterium vector and the carrier that can be used for the plant micropellet bombardment etc.Utilize any one carrier that can guide the outer rim gene in plant, to express, OsWTF1 gene of the present invention overexpression in plant is shown characteristics of drought tolerance.
Gene of the present invention can add any one strong promoter or inducible promoter on being building up to plant expression vector the time before its transcription initiation Nucleotide, also can use enhanser.
Carry OsWTF1 expression carrier of the present invention and can change vegetable cell over to, and cultivate the plant that drought resistance obtains improveing through using conventional biotechnological meanss such as Ti-plasmids, Ri plasmid, plant viral vector, directly DNA conversion, microinjection and electroporation.By the plant transformed host can be monocotyledonss such as paddy rice, corn, wheat, also is applicable to dicotyledonss such as tobacco, soybean, cultivates the plant variety of drought resisting.
Below in conjunction with accompanying drawing and specific embodiment the present invention is explained further details.
Description of drawings
Fig. 1 adopts ClustalW software (public use software) that homologous protein in rice Os WTF1 albumen and its Arabidopis thaliana is carried out the homology comparative result.
Wherein: black is represented the amino acid of high conservative.
Fig. 2 is the conservative domain analysis figure of OsWTF1 gene coded protein.
Fig. 3 is the full length cDNA clone of OsWTF1
From left to right be followed successively by among the figure: M, molecular weight size Marker; 1,5 ' RACE amplified production; The negative control of 2,5 ' RACE amplification; 3,3 ' RACE amplified production; The negative control of 4,3 ' RACE amplification.5 ' RACE amplification obtains the fragment about about 750bp; 3 ' RACE amplification obtains the fragment about about 300bp; 5 ' RACE and 3 ' RACE product are cut the glue recovery, and change pGEM-T Easy Vector over to, obtain the 791bp full-length cDNA through the order-checking splicing; Coding region length 618bp, predictive coding protein are 205aa.Clone's full-length cDNA Genbank login (DQ468387, ABE98440).
Fig. 4 is the gene expression analysis figure of OsWTF1 under arid and the uv-radiation condition.
Wherein: CK representes OsWTF1 expression of gene in the fine blade of untreated Japan, and 1 expression arid is handled OsWTF1 expression of gene in the rice leaf of back, under the 2 expression ultraviolet treatment condition, and OsWTF1 expression of gene in the rice leaf.The OsActin expression of gene is used as confidential reference items, and (expression one of OsActin is taken turns, 28cycles).
Fig. 5 is PCR and the RT-PCR detected result figure of OsWTF1 overexpression transgenic plant.
Wherein: the PCR of A:OsWTF1 overexpression transgenic plant detects.The plasmid that P representes to contain the OsWTF1 gene fragment is as positive control; 1,2 contrasts as recessiveness for the non-transgenic plant; 3-16 is that independent transfer-gen plant strain is.
The RT-PCR of B:OsWTF1 overexpression transgenic plant detects.P representes to contain the plasmid of OsWTF1 gene fragment; CK1 and CK2 are the non-transgenic adjoining tree, and all the other are independent transgenic line, and the OsActin gene is used as confidential reference items.
Fig. 6 is OsWTF1 overexpression rice plant blade epicutile wax crystalline scanning electron microscopic observation result.
Wherein: a
1Be the wild-type adjoining tree blade outside of belly; a
2For being wild-type adjoining tree vacuum side of blade; b
1Be the OsWTF1 overexpression transgenic paddy rice blade outside of belly; b
2Be OsWTF1 overexpression transgenic paddy rice vacuum side of blade.
Fig. 7 is the chlorophyll extraction rate analysis of OsWTF1 overexpression rotaring gene plant blade.
Fig. 8 is the resistance reaction and display figures of OsWTF1 transgenic plant to arid.Be the rice plant in tillering phase that is incubated at the greenhouse to be carried out drought resisting handle experiment.
Wherein: a is the Japanese fine plant of wild-type (WT), and b is the OsWTF1 transfer-gen plant.
Be respectively like figure: before arid is handled, OsWTF1 overexpression transgenic paddy rice b
1With wild-type a
1Comparison;
After arid is handled a week, OsWTF1 overexpression transgenic paddy rice b
2With wild-type a
2Comparison;
Arid is handled after a week one week of rehydration again, OsWTF1 overexpression transgenic paddy rice b
3With wild-type a
3Comparison.
Embodiment
Experimental technique among the following embodiment like no specified otherwise, is ordinary method.
The acquisition of embodiment 1, plant adversity resistance related protein OsWTF1 and encoding sox thereof
The result that little display (Microarray) is analyzed shows that rice Os 02g0202000 gene is the transcription factor gene of an AP2 type, the ERF type paddy rice wax metabolism transcription factor of encoding and containing " AP2 " structural domain.Utilize blast program find it with Arabidopis thaliana in WIN1 height homology, the research of WIN1 is shown that it is a transcription factor of participation wax pathways metabolism.So we are OsWTF1 with the Os02g0202000 unnamed gene.According to BAC library AP004869 sequences Design 5 ' RACE amplimer: WTF1R1 and the WTF1XbaR that paddy rice has been announced, 3 ' RACE amplimer: WTF1F1 and WTF1BamF carries out the full-length cDNA amplification of OsWTF1 gene.
Amplification condition is with reference to SMART
TMRACE cDNA Amplification Kit explanation is operated.
5 ' RACE: under the effect of Superscript III reversed transcriptive enzyme, be that template, 5 ' CDS primer and SMARTII A oligo are primer with the total RNA of 4 μ L rice young panicles, the strand cDNA of synthetic 5 ' RACE.
3 ' RACE: under the effect of Superscript III reversed transcriptive enzyme, total RNA is a template with 4 μ L rice young panicles, adds 3 ' CDS primer A and DTT, dNTP and First-strand Buffer, and 42 ℃ add 1.5h, the strand cDNA of synthetic 3 ' RACE.
After 5 ' RACE strand cDNA and 3 ' RACE strand cDNA are synthetic, 100 times of dilution reverse transcription products, 5 ' RACE is a template with 5 ' RACE strand cDNA, is primer with UPM and WTF1R1; 3 ' RACE is a template with 3 ' RACE strand cDNA, is that primer carries out touchdown PCR, reaction conditions with UPM and WTF1F1: 94 ℃ of 5s, 72 ℃ of 3min, 5 circulations; 4 ℃ of 5s, 70 ℃ of 10s, 72 ℃ of 3min, 5 circulations; 94 ℃ of 5s, 68 ℃ of 10s, 72 ℃ of 3min, 25 circulations; 4 ℃ of preservations.First round PCR product is carried out the PCR reaction with UPM, WTF1XbaR and UPM, WTF1BamF respectively, and amplification condition is 94 ℃ of 3min; 94 ℃ of 30s, 60 ℃ of 30s, 72 ℃ of 1min, 35 circulations; 72 ℃ of 5min; 4 ℃ of preservations.
The PCR product is carried out agarose gel electrophoresis to be detected; Reclaim test kit with sepharose and reclaim 5 ' RACE and 3 ' RACE fragment; To reclaim fragment is connected with carrier pGEM-T Easy; To connect product transformed into escherichia coli DH5 α competent cell,, obtain containing the segmental recombinant plasmid of recovery according to the carboxylic Bian penicillin resistance label screening positive colony on the pGEM-T Easy carrier.With T7 on this recombinant plasmid vector and SP6 promoter sequence is that primer carries out nucleotide sequencing to it; In conjunction with Fig. 3; Sequencing result shows the protein-coding region fragment of increased OsWTF1 gene 5 ' end and 3 ' end, and then from 2 the full-length cDNA that obtains total length OsWTF1 gene 3 ' of overlapped sequence-RACE and 5 '-RACE product is arranged.The encoding histone region sequence of OsWTF1 gene is made up of 618 deoxyribonucleotides shown in SEQ No:3, the albumen shown in the coding SEQ No:2.
Auele Specific Primer information is following:
Upstream primer WTF1BamF 5 '-AT
ATCTCTGCCTCCCCTGTCCTTC-3 '; The base of overstriking band underscore is the BamHI restriction enzyme site
Downstream primer WTF1XbaR 5 '-AT
CGTCGTTTCATCCAAGCCTTTC-3 ', the base of overstriking band underscore is the XbaI enzyme cutting site;
Upstream primer WTF1F1:5 '-GACTCCAACTGGGTGATGACGGTG-3 ',
Downstream primer WTF1R1:5 '-TTGCTACGGGCGTCCTGGTCTGC-3 '.
Implement the Blast comparison of row 2, OsWTF1 gene, the Subcellular Localization of proteins encoded and conserved structure domain analysis
Referring to Fig. 1 and Fig. 2, CLUSTAL W and DNAMAN software through http://www.ncbi.nlm.nih.gov/BLAST/ website carry out homology analysis.The homology of rice Os WTF1 gene and Arabidopis thaliana wax metabolism transcription factor gene WIN1 (SHN1) is higher; OsWTF1 and the amino acid whose homology of arabidopsis gene WIN1 (at1g15360) are 61%; With the amino acid identity of At5g25390 be 54%, with the amino acid identity of At5g11190 be 55%.Arabidopsis gene WIN1 (SHN1, at1g15360), At5g11190, At5g25390 gene be for regulation and control wax metabolism related gene, infers that the OsWTF1 gene maybe be relevant with the metabolism of paddy rice wax.Through http://wolfpsort.org/ website Subcellular Localization analysis software, finding with the higher known of OsWTF1 homology has 11 to be positioned in the nucleus, and 2 are positioned in the chloroplast(id).Preliminary definite, the albumen of OsWTF1 genes encoding is positioned in the nucleus.Sequence conserved regions according to rice Os WTF1 gene coded protein is carried out functional analysis with the software that http://www.ncbi.nlm.nih.gov/Structure/cdd/wrpsb.cgi provides; And carry out protein structure prediction with http://swissmodel.expasy.org/ software; Find that OsWTF1 has an AP2 structural domain; At the 6-66aa place, belong to the transcription factor of EREF class.
Be the reaction of check OsWTF1 gene under adverse environmental factor, adopt arid and uv-radiation method processing rice leaf also to detect and handle OsWTF1 expression of gene in the plant of back.
The paddy rice of conventional cultivation to late tillering state, be divided into three groups, one group of contrast, growth under natural light; The ultraviolet treatment group adds the UVB pipe in addition under the natural lighting condition provide UVB radiation (15W, Shanghai Gu Cun photoelectric instrument factory).Ultraviolet lamp tube is positioned plant upper end 20-30cm, and UVB radiation photophase 8h/d (treatment time is 9:00-17:00 every day) handles week age.The arid treatment group then under natural light growth add 6 days arids in addition and handle.Get blade after the processing and adopt the Trizol extract of Invitrogen company to extract total RNA, and to be template handle in the rear blade OsWTF1 expression of gene to adverse circumstance carries out RT-PCR and analyze with total RNA.Reverse transcription system: 2 μ L 50ng/ μ L Olig (dt); 4.0 μ L 5X first strand buffer, 0.4 μ L 100mmol/L DDT, 2.0 μ L 10mmol/L dNTP, 1.0 μ L M-MLV 200U/ μ L add H2O to the 20 μ L that DEPC handles again.In 37 ℃ of water-bath 1h.Place 95 ℃ of following 5min, deactivation M-MLV adds 2 μ L TE (pH=8.0), synthesizing single-stranded cDNA.With each cDNA first chain of synthetic is template, detects the OsWTF1 expression of gene with WTF1BamF and WTF1XbaR.Find that under the normal cultivation condition, the expression of OsWTF1 gene in rice leaf is lower, just see the band of expression behind the cDNA process two-wheeled PCR (first round 28cycles second takes turns 30cycles) of reverse transcription.
Be used as confidential reference items with the OsActin gene, reaction conditions is: 94 ℃ of preparatory sex change 3min; 94 ℃ of 30sec, 60 ℃ of 30sec; 72 ℃ of 35sec, 28 circulations; 72 ℃ are extended 10min.Its Auele Specific Primer is following:
ActinBamF：5’-ATGGATCCAGGTAATAAGATCTTCAACAC-3’
ActinSpeR：5’-ATACTAGTACCTGGAAACACACAAGACA-3’
As shown in Figure 4, after ultraviolet and the arid of for some time were handled, the OsWTF1 expression of gene all increased than illumination is apparent in the rice leaf, and arid is handled the increase ratio that OsWTF1 is expressed and handled more remarkable through ultraviolet.
Implement the structure and the conversion of row 4, OsWTF1 gene overexpression carrier
In order to illustrate the function of OsWTF1 gene better, the applicant carries out the functional verification of OsWTF1 gene with its overexpression in paddy rice from the phenotype of transgenic plant.
The overexpression carrier construction method is following: total length OsWTF1 full-length cDNA (SEQ ID No:3) directed cloning to pCambia1300M carrier 35S promoter downstream, is used to make up the plant binary expression vector of paddy rice 35S-OsWTF1 overexpression.After freeze-thaw method changes among the Agrobacterium EHA105; Through agriculture bacillus mediated rice genetic method for transformation it is imported in the rice varieties " Japan is fine "; Through the callus of cultivating in advance, infecting, cultivating altogether, screening having hygromycin resistance, break up, take root, acclimatization and transplants, obtain transfer-gen plant.Agriculture bacillus mediated rice genetic method for transformation is used people's reported method (Qu et al., 2006) such as Qu.
Implement the Molecular Detection of row 5, OsWTF1 gene overexpression transgenic plant
OsWTF1 gene overexpression transgenic Molecular Detection of the present invention adopts PCR and RT-PCR method to carry out.
5.1OsWTF1 the PCR Molecular Detection on the gene overexpression transfer-gen plant DNA aspect
Extract oryza sativa genomic dna with the CTAB method: the about 100mg of clip rice leaf with behind the liquid nitrogen pulverize, adds 600 μ L DNA extraction damping fluids of 60 ℃ of preheatings in mortar, 60 ℃ of water bath heat preservation 30-60min, and gentleness is shaken frequently.Add 600 μ L equal-volume chloroform/primary isoamyl alcohol, put upside down mixing, leave standstill 5-10min under the room temperature, make water and organic phase layering.The centrifugal 30min of 10000r/min under the room temperature.Get supernatant to another 1.5mL eppendorf pipe, add the long-pending Virahol of monoploid, mixing, the centrifugal 10min of 10000r/min behind-20 ℃ of placement 30min; 70% alcohol washes twice, dries, and adds the dissolving of 100 μ L TE solution.
The PCR of OsWTF1 gene overexpression transfer-gen plant identifies:
Get commentaries on classics OsWTF1 rice plant genomic dna 100ng with hygromycin resistance; Carry out pcr amplification with HygF and HygR primer and identify the Totomycin encoding sox; Carrying out pcr amplification with primer WTF1BamF and WTF1XbaR identifies; The positive contrast of expression plasmid was set simultaneously, the not negative contrast of transfer-gen plant genomic dna.The result is shown in A among Fig. 5; All transgenic positive plant can both amplify the dna fragmentation and the cDNA fragment that is inserted into the about 700bp of OsWTF1 total length in the rice genome of about 820bp band intron that rice genome itself has; And the negative plant of not genetically modified negative control plant and the transgenic dna fragmentation that the rice genome self of about 820bp band intron has that all can only increase, plasmid contrasts and also can expand the OsWTF1 full-length cDNA fragment that about 700bp.PCR detects positive plant 22 strains that obtain to change the OsWTF1 full-length cDNA.
5.2OsWTF1 the RT-PCR Molecular Detection on the gene overexpression transfer-gen plant RNA aspect
Extract the total RNA of the fine young fringe of young fringe total RNA of portion and not genetically modified Japan of test positive plant on the dna level; It is template that reverse transcription becomes cDNA; Use WTF1BamF and WTF1XaR to carry out PCR as primer; The expression of analysis purposes gene on transcriptional level, and with the ACTIN expression of gene as confidential reference items.The method of the extraction of RNA and RT-PCR reaction is with the RT-PCR method of implementing in the row 3.The result compares with transfer-gen plant not shown in B among Fig. 5, and OsWTF1 has obtained the enhancing of different expression degree and expressed in the part transfer-gen plant, and wherein No. 6 transfer-gen plants obtain strongly expresseds, can be used for further phenotypic evaluation.
Implement the scanning electron microscopic observation of row 6, OsWTF1 gene overexpression rotaring gene plant blade surface wax
Get same vegetative period overexpression change the OsWTF1 gene plant and not the paddy rice of genetically modified adjoining tree open up the leaf blade entirely and fix with LUTARALDEHYDE; Be stored in 4 ℃ the refrigerator; After the requirement dehydration of ESEM, drying; With sample metal spraying plated film on JEOL JFC-1600 coating equipment, under the JSM-6360LV ESEM, observe and take pictures.The result is as shown in Figure 6: the OsWTF1 overexpression plant leaf back side (b2) and the outside of belly (b1) have all that epicutile wax obviously thickens, mastoid process increases increase, the mastoid process form changes; The mastoid process height obviously increases; Become near modal variations such as cylindrical shapes, and these change in the outside of belly (b1) performance of changeing OsWTF1 trans-genetic hybrid rice blade more obvious.
Implement the chlorophyll extraction rate of row 7, OsWTF1 gene overexpression rotaring gene plant blade
The chlorophyll extraction rate that changes OsWTF1 gene plant blade is undertaken by the per-cent that certain hour inner chlorophyll lixiviate amount accounts for the chlorophyll total amount.
Get OsWTF1 gene overexpression transfer-gen plant and adjoining tree (not genetically modified wild-type plant) first functional leaf under same heading stage sword-like leave; Weigh after the tap water flushing; Respectively get 5g and put into triangular flask, add the ethanol of 30mL 80%, place under the dark condition and shake once in a while.Preceding 1h is every to get 400 μ L vat liquors at a distance from 10min from each sample, respectively get 400 μ L vat liquors again at 90min, 120min then, measures its absorption value under λ 664nm and λ 647nm wavelength.By formula calculate chlorophyll total amount (mmol/g)=7.93A664+19.53A647.The result is as shown in Figure 7, and As time goes on, transgenic improves with the chlorophyll extraction rate of contrast wild-type plant gradually; But in all processes of lixiviate; Change the leaf chlorophyll extraction rate of OsWTF1 gene plant and all compare according to low always, behind the alcohol lixiviate 120min, about 80% chlorophyll is not come out by lixiviate in the transgenic wild-type plant leaf; And the blade extraction rate that changes the OsWTF1 gene plant shows that less than 40% transfer-gen plant reduces chlorophyllous permeability.
The reaction that enforcement row 8, OsWTF1 gene overexpression transgenic plant are handled arid
As a result under the prerequisite, the present invention has at first carried out the homozygote screening to OsWTF1 gene overexpression transgenic plant T3 for plant at embodiment 5.Concrete steps are following: each plant is chosen 30 seeds and shells, 70% ethanol surface sterilization 1min, 30% sodium hypochlorite solution sterilization 30min; Sterilization washing 5 times; Containing the 1/2MS of Totomycin (Murashige & Skong) the enterprising row filter of substratum plate with the aseptic nipper sowing, wild-type Japan is fine as contrast, and culture condition is 28 ℃; Continuous light intensity is 2500lux, and 16h illumination/8h is dark.Back statistics germination of two weeks and surviving rate, the plant that the present invention gives tacit consent to 100% surviving rate is a homozygote, each homozygote plant is taken a sample separately and carries out corresponding Molecular Detection.Get the Molecular Detection strain system consistent and be used for ensuing phenotypic evaluation with the hygromycin selection result.
The present invention choose three independently the pure and mild paddy rice of overexpression transgenic be used for phenotypic screen.Get pure and mild transgenic line seed and wild-type fine seed germination of Japan and routine and be cultured to mid-term in tillering phase, do not water and carry out the arid processing.
After one week of arid, as shown in Figure 8, OsWTF1 overexpression plant is keeping normal growth conditions (b basically
2), and contrast wild-type (a
2) here great majority wither, blade is sagging, serious drought occurs and coerces performance.
In one week back one week of rehydration of arid, as shown in Figure 8, most contrast wild-type plant blades occur expendable withered, only have two of minority to tiller and turn green, and see a
3And OsWTF1 overexpression plant all can return to the normal growth state, sees b
3The rehydration ability of plant after arid also is an important indicator of drought resisting.Arid and rehydration experiment show that the OsWTF1 overexpression has strengthened the drought tolerance of plant, and the result confirms that OsWTF1 has play a part important to the response of arid.
Claims (8)
1. albumen is (a) or protein (b) as follows:
(a) protein of forming by the aminoacid sequence shown in the SEQ ID No:2 in the sequence table;
(b) with the aminoacid sequence of SEQ ID No:2 in the sequence table through the replacement of one or several amino-acid residue and/or disappearance and/or interpolation and relevant with stress resistance of plant by (a) deutero-protein.
2. albumen according to claim 1 is characterized in that: the replacement of said one or several amino-acid residue and/or disappearance and/or interpolation are meant that the 1st amino acids to the 206 amino acids at SEQ ID No:2 replace and/or lack and/or add.
3. claim 1 or 2 said proteic encoding soxs.
4. gene according to claim 3 is characterized in that: (a) or (b) or gene (c) as follows:
(a) its nucleotides sequence is classified SEQ ID No:1 in the sequence table as from the dna molecular shown in 5 ' terminal the 1st to the 912nd deoxyribonucleotide,
(b) its nucleotides sequence is classified the dna molecular shown in the SEQ ID No:1 in the sequence table as,
The dna molecular of the code for said proteins of (c) under stringent condition, hybridizing with the dna sequence dna that (a) limits.
5. the recombinant expression vector that contains claim 3 or 4 said genes.
6. recombinant expression vector according to claim 5; It is characterized in that: said recombinant expression vector is pCambia1300M-OsWTF1, and said pCambia1300M-OsWTF1 inserts the recombinant expression vector that claim 3 or 4 said genes obtain at the MCS of pCambia1300M carrier.
7. the transgenic cell line or the reorganization bacterium that contain claim 3 or 4 said genes.
8. a method of cultivating the drought resisting paddy rice is that claim 5 or 6 described recombinant expression vectors are imported in the rice cell, obtains the drought resisting paddy rice.
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| CN107326033B (en) * | 2017-08-28 | 2019-08-09 | 中国科学院东北地理与农业生态研究所 | Application of the OsROC4 gene in the synthesis of adjusting and controlling rice epicutile wax |
| CN114605513A (en) * | 2022-01-24 | 2022-06-10 | 国际竹藤中心 | Phyllostachys pubescens waxy synthesis transcription factor gene PeWST and application thereof |
| CN114605513B (en) * | 2022-01-24 | 2023-12-22 | 国际竹藤中心 | Moso bamboo waxy synthetic transcription factor gene PeWST and application thereof |
| CN114561404A (en) * | 2022-04-20 | 2022-05-31 | 河南农业大学 | Apple MdSHN1 gene and application thereof in improving waterlogging tolerance of plants |
| CN114561404B (en) * | 2022-04-20 | 2023-06-16 | 河南农业大学 | Apple MdSHN1 gene and application thereof in improving waterlogging tolerance of plants |
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