CN1687124A - Drought resistant correlative protein and coded gene of plant and application - Google Patents
Drought resistant correlative protein and coded gene of plant and application Download PDFInfo
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- CN1687124A CN1687124A CN 200510066436 CN200510066436A CN1687124A CN 1687124 A CN1687124 A CN 1687124A CN 200510066436 CN200510066436 CN 200510066436 CN 200510066436 A CN200510066436 A CN 200510066436A CN 1687124 A CN1687124 A CN 1687124A
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Abstract
The present invention discloses a plant drought-resistance related protein, its coded gene and application. The coded gene of said plant drought-resistance related protein can be used to raise the drought resistance of plant, for example it can be used for inhibiting expression of the above-mentioned plant drought-resistance related protein coded gene in the plant. The invented plant drought-resistance related protein and its coded gene can be used for breeding new variety of drought-resistant plant.
Description
Technical field
The present invention relates to a kind of plant drought associated protein and encoding gene and application in the bioengineering field, particularly utilize this gene to strengthen the method for plant drought resistance.
Background technology
Food problem is one of several hang-ups of facing of the world today.By improve per unit area yield or enlarge cultivated area, the approach such as improve the low and medium-yield farmland of increasing investment in agriculture increases grain yield and all can run into and need overcome the adverse circumstance restriction or alleviate the problem of adverse circumstance harm.Therefore, the understanding plant is improved the resistance of plant to the reaction mechanism of adverse circumstance, has become the further important foundation research of volume increase of agricultural, and extremely countries in the world government and scientist's concern also is the focus of current life science.
Arid is the main adverse circumstance that limiting plant growth is grown, destruction along with the mankind's Economic development, population expansion and biotic population, the shortage of water resources phenomenon is on the rise, it has seriously influenced agriculture production and ecotope, become the problem that the whole world is paid close attention to, therefore seek plant particularly farm crop drought resisting approach be very necessary.Improve the drought resistance of crop, except utilizing traditional breeding method, at present, one of field that the using gene engineering breeding has become the scientific worker to be paid close attention to.
Arabidopis thaliana is a kind of typical model plant, is widely used in plant genetics, developmental biology and molecular biological research.Most of genes of Arabidopis thaliana can both find in other plant, and any discovery of relevant Arabidopis thaliana can both be applied to other plant research.Therefore, the research to Arabidopis thaliana drought resistance The Molecular Biology Mechanism will greatly help to find raising farm crop drought resistance, the method for increase output.
Arabidopis thaliana has about 1.3 hundred million base pairs, 2.9 ten thousand genes.The function of most gene is not clear at present, utilizes mutating technology research gene function to become a kind of effective means.By research, known the function of some anti-drought genes, as DREB, CBF, ABRE etc. to mutant.
At present, also do not find effective drought resisting functional gene,, seek new drought resisting functional gene and illustrate its function and have important theory and practice significance in the face of serious day by day crop arid problem.
Summary of the invention
The purpose of this invention is to provide a kind of new plant drought associated protein and encoding gene thereof.
Plant drought associated protein provided by the present invention, name is called LEW2-1 (LEaf Wilt 2-1), and it derives from the ecotypic Arabidopis thaliana of Colombia, is the protein with one of following amino acid residue sequences:
1) the SEQ ID № in the sequence table: 2;
2) with SEQ ID № in the sequence table: 2 amino acid residue sequence is through replacement and/or disappearance and/or the interpolation and the protein relevant with plant drought of one or several amino-acid residue.
Sequence 2 in the sequence table is made up of 985 amino-acid residues.
The replacement of described one or several amino-acid residue and/or disappearance and/or interpolation are meant replacement and/or disappearance and/or the interpolation that is no more than 10 amino-acid residues.
The encoding gene of LEW2-1 (LEW2-1) also belongs to protection scope of the present invention.
The cDNA gene of LEW2-1 can have one of following nucleotide sequence:
1) SEQ ID № in the sequence table: 1 dna sequence dna;
2) SEQ ID № in the code sequence tabulation: the polynucleotide of 2 protein sequences;
3) under the rigorous condition of height can with SEQ ID № in the sequence table: the nucleotide sequence of the 1 dna sequence dna hybridization that limits;
4) with sequence table in SEQ ID №: 1 dna sequence dna that limits has 90% above homology, and the identical function protein DNA sequence of encoding.
The rigorous condition of described height can be 0.1 * SSPE (or 0.1 * SSC), in the solution of 0.1%SDS, under 65 ℃, hybridize and wash film.
Sequence 1 in the sequence table is by 3217 based compositions, its open reading frame (ORF) be from the 68th at 5 ' end to 3025 bit bases, the protein of encoding sequence 2.
The expression vector that contains LEW2-1 of the present invention, clone and host bacterium all belong to protection scope of the present invention.Arbitrary segmental primer is to also belonging to protection scope of the present invention among the amplification LEW2-1.
Second purpose of the present invention provides a kind of method of utilizing this gene to strengthen plant drought resistance.
The method of enhancing plant drought resistance provided by the present invention is the expression that suppresses the above-mentioned plant drought resistance associated protein encoding gene in the plant.
Suppress the expression of the above-mentioned plant drought resistance associated protein encoding gene LEW2-1 in the plant and can pass through accomplished in many ways, method as the plant viral vector mediated gene silencing, the method of antisense technology silencer, the gene silencing methods of siRNA mediation etc.The method of inhibition of gene expression of the present invention is not limited to above-mentioned several method, all can as long as can suppress the LEW2-1 expression.
Utilize any plant gene to knock out technology, behind this gene knockout (or reticent), plant shows as drought resisting; Utilize any carrier that can guide foreign gene to express in plant, LEW2-1 provided by the present invention is changed in the plant, it is responsive that plant just shows drought.
LEW2-1 gene of the present invention or its antisense nucleic acid can add any enhancing promotor or inducible promoter in being building up to plant expression vector the time before its transcription initiation Nucleotide.For the ease of transgenic plant cells or plant being identified and screening, can process employed carrier, as the antibiotic marker thing (gentamicin, kantlex etc.) that adds the alternative mark (gus gene, luciferase genes etc.) of plant or have resistance.By the plant transformed host both can be monocotyledons, also can be dicotyledons, as: paddy rice, wheat, corn, cucumber, tomato, willow, turfgrass or lucerne place etc.From the security consideration of transgenic plant, can not add any selected marker, directly screen transformed plant with plant seedling leaf dehydration degree.Carry that LEW2-1 expression carrier of the present invention can Ti-plasmids, Ri plasmid, plant viral vector, directly DNA conversion, microinjection, electricity be led, conventional biological method transformed plant cells or tissue such as agriculture bacillus mediated by using, and plant transformed is become plant through tissue cultivating.
Plant drought associated protein of the present invention and encoding gene thereof provide the support (guarantee) of gene and technology for farm crop drought resisting breeding.
Below in conjunction with embodiment technical scheme of the present invention is further described.
Description of drawings
Fig. 1 is that the drought resistance of lew2-1, wild-type plant compares photo
Fig. 2 is that the osmotic adjustment ability of lew2-1, wild-type plant compares photo
Fig. 3 is the expression characteristic of lew2-1, wild-type plant adversity gene under normal and environment stress condition
Embodiment
Experimental technique among the following embodiment is ordinary method if no special instructions.
The acquisition of embodiment 1, LEW2-1 and encoding gene thereof
Seedling 100-200mg with the environmental Arabidopis thaliana of wild-type Colombia is a material, extracts root and the total RNA of stem respectively with Trizol, detects the integrity of RNA through 1% agarose electrophoresis.Ss cDNA's is synthetic according to SuperScript
TMThe method of II RNase H-Reverse Transcriptase, synthesizing single-stranded (ss) cDNA.With 10 times of synthetic strand cDNA dilutions, template as following PCR reaction: 20 μ l systems include 10 * PCR damping fluid, 2 μ l, 2.5mM dNTP (dATP, dGTP, dCTP, dTTP) mixture 1.6 μ l, each 1.0 μ l of the primer 1 of 5 μ m and primer 2, Taq enzyme (15U/ μ l) 0.1 μ l.Wherein, primer 1:5 ' ATGATGGAGTCTAGGTCTCCCAT 3 ', primer 2: 5 ' ACTGTCTT GATCGTTATTGCTAA3 '.On PE 9700 instrument, increase: 94 ℃ of pre-sex change 3min of elder generation; 94 ℃ of 30sec again, 55 ℃ of 30sec, 72 ℃ of 1min 30sec amount to 35 circulations; Last 72 ℃ are extended 4min, with the PCR product that obtains, make 1% agarose gel electrophoresis.The cDNA fragment that PCR is obtained is connected in the pGEM-T carrier, obtain containing the segmental carrier pT-LEW2-1 of purpose, the evaluation of checking order, the result shows the dna sequence dna that cDNA fragment that PCR obtains has sequence 1 in the sequence table, cDNA gene for LEW2-1, by 3217 based compositions, its encoding sequence is that coding has the protein of the amino acid residue sequence of sequence 2 in the sequence table from 5 ' end the 68th bit base to the, 3025 bit bases.
Embodiment 2, cultivation drought resistance enhanced Arabidopis thaliana
1, the acquisition of the homozygous mutation body lew2-1 that knocked out of LEW2-1 gene
According to the specific fragment in the LEW2-1 gene order, design primer: RNAi-F:5 ' ATGATGGAGTCTAGGTCTCCCAT 3 ' and RNAi-R:5 ' ACTGTCTTGATCGTTATTGCTAA 3 ', with the environmental arabidopsis thaliana genomic dna of Colombia is template, carry out pcr amplification, obtain the purpose fragment that length is about 350bp, (entry vector makes up conversion system to utilize the gateway system, available from INVITROGEN company) this 350bp purpose fragment is incorporated among the plasmid pJawoh-8 (available from INVITROGEN company), form inverted repeats, correct through order-checking detection integration fragment and direction of insertion, (selection markers is a kantlex to transform Agrobacterium, penbritin, gentamicin), utilize Agrobacterium to infect the method arabidopsis thaliana transformation, the F1 that obtains is used weedicide grass fourth phosphorus (basta for plant, available from INVITROGEN company) screening conversion seedling, separation and purification.To not having the strain that separates the proterties phenomenon through careless fourth phosphorus screening is that Trizol extracts total RNA, changes nylon membrane after the agarose electrophoresis, carries out Northern-blot and detects.Wherein, used probe is for being template with wild-type Arabidopis thaliana group DNA, utilize primer 1 and primer 2 to carry out the dna fragmentation that pcr amplification obtains, wherein 20 μ l PCR reaction systems include 10 * PCR damping fluid, 2 μ l, 2.5mM dNTP (dATP, dGTP, dCTP, dTTP) mixture 1.6 μ l, each 1.0 μ l of the primer 1 of 5 μ M and primer 2, Taq enzyme (15U/ μ l) 0.1 μ l.On PE 9700 instrument, increase: 94 ℃ of pre-sex change 3min; 94 ℃ of 30sec, 55 ℃ of 30sec, 72 ℃ of 1min 30sec amount to 35 circulations; 72 ℃ are extended 4min, with the PCR product that obtains, make 1% agarose gel electrophoresis.With total RNA hybridization, the result does not obtain hybridizing band, proves that the RNA of LEW2-1 genetic expression is degraded, obtains homozygous mutation body lew2-1 behind the probe P32 mark.
2, the drought resistance of lew2-1, wild-type plant relatively
Lew2-1 plant, the wild-type plant in 2 weeks of will growing in soil stopped respectively watering 10-14 days, and other culture condition is all identical, and here situation is to determine its drought resistance to observe that relatively plant survival rate and blade are withered.The result as shown in Figure 1, the survival rate that shows wild-type plant (WT) is 2%, the survival plant 1/3rd blade withered and yellow; The lew2-1 plant that the LEW2-1 gene is knocked out all survives, and does not have the withered and yellow blade of wilting; After LEW2-1 genetically deficient is described, can make the plant drought resisting,
3, the osmotic adjustment ability of lew2-1, wild-type plant relatively
The seed of lew2-1, wild-type plant is sowed respectively on common (MS) substratum, cultivated 5-6 days under the dark condition in 8 hours illumination in 22 ℃, 16 hours after following vernalization 2-3 days for 4 ℃, transfer to then on the substratum of the seminose that contains different concns (0,420,520mmol/L) and cultivated 7-10 days under the dark condition in 8 hours, observe relatively plant root and whole strain growing state to determine their osmotic adjustment ability illumination in 22 ℃, 16 hours.The result shows the rising along with mannose concentration as shown in Figure 2, and the root growth of wild-type plant (WT) slows down, and changes obviously; When mannose concentration reaches 420mmol/L, all 3/4ths blade flavescence of plant, when mannose concentration reaches 520mmol/L, all whole blade flavescence of plant; Though the root growth of the lew2-1 plant that the LEW2-1 gene is knocked out also slows down, change being not so good as the obvious of wild-type plant, when mannose concentration reaches 520mmol/L, have only 1/3rd blade flavescence of 1/5th plant; After LEW2-1 genetically deficient was described, the plant osmotic adjustment ability strengthened, and helps drought resisting.
Adversity gene expression in embodiment 3, detection lew2-1, the wild-type plant
Be chosen at the lew2-1 in 2 weeks of growth in common (MS) substratum, the wild-type plant carries out following processing respectively:
Water planting is handled: 2 weeks of growth in common liq (MS) substratum;
Arid is handled: at room temperature expose and placed 3 hours;
High optical processing: in common (MS) substratum under high light intensity (260 micromoles/second square centimeter) condition 2 weeks of growth;
Extract the plant RNA of above-mentioned processing respectively, be that (used probe is to be template with the plant genomic dna in contrast with rRNA, utilize primer ATGATGGAGTCTAGGTCTCCCAT and ACTGTCTTGATCGTTATTGCTAA to carry out the dna fragmentation that PCR obtains), utilize the RNA blot hybridization technique to detect adversity gene: Arabidopis thaliana arid, high salt and low temperature response gene RD29A, drought-enduring gene P5CS of anti-salt and ABA (dormin) synthetic gene SDR1, expression in lew2-1, wild-type plant respectively is to determine that it is to corresponding resistance size of coercing.Wherein, detecting the used probe of RD29A and be with the plant genomic dna is template, utilizes primer 5 ' GACGAG TCAGGA GCTGAGCTG3 ' and 5 ' CGATGCTGCCTTCTCGGTAGAG3 ' to carry out the dna fragmentation that PCR obtains; Detecting the used probe of P5CS and be with the plant genomic dna is template, utilizes primer 5 ' AGCCTTGGCACAGGAGCAACG3 ' and 5 ' TGAGAC CAGTGACAGCATCAAACA3 to carry out the dna fragmentation that PCR obtains; The used probe of SDR1 is to be template with the plant genomic dna, utilizes primer 5 ' TGATCACTGGAGGAGCCACAGG 3 ' and 5 ' CTCCGCTTATGTACC GCGAGTC3 ' to carry out the dna fragmentation that PCR obtains.
The result shows lew2-1 plant (control) under normal operation as shown in Figure 3, and water planting, under the arid and high optical processing condition, and RD29A, the expression amount of P5CS and SDR1 all is significantly higher than the wild-type plant.After LEW2-1 genetically deficient is described, can make the plant drought resisting, among Fig. 3, W represents the wild-type plant, and M represents the lew2-1 plant.
Sequence table
<160>2
<210>1
<211>3217
<212>DNA
＜213〉Arabidopsis Arabidopis thaliana (Arabidopsis thaliana)
<400>1
atccatccaa?atctcaatcc?ctaattaggg?ttcatttctc?tgtttctcca?aacaggggaa 60
ttcgaagatg?atggagtcta?ggtctcccat?ctgcaacact?tgtggtgaag?agattggtgt 120
aaaatcaaac?ggagagttct?ttgtggcttg?tcatgagtgt?agtttcccga?tctgcaaagc 180
ttgtcttgag?tatgaattca?aagaaggtcg?aagaatttgc?ttgcgttgcg?gcaatcctta 240
cgatgagaat?gtgtttgatg?atgttgagac?aaagacatct?aaaactcaat?ccattgttcc 300
aacacagacc?aataacactt?ctcaggattc?agggattcat?gctagacata?taagtacagt 360
ctcaacaata?gacagtgaac?tgaatgatga?atatggcaat?ccaatttgga?agaacagagt 420
ggagagctgg?aaagacaaga?aagacaagaa?gagcaagaag?aagaagaaag?atccaaaagc 480
aacaaaagct?gaacaacatg?aggctcagat?tcctacccaa?cagcacatgg?aagatacgcc 540
accgaacaca?gaatctggtg?ctacagatgt?gctttcggtt?gtgattccta?tcccaaggac 600
aaaaatcact?tcatatagga?ttgtcatcat?catgcggttg?atcatcttgg?ctctgttttt 660
taactaccgt?atcacgcatc?ctgtcgatag?cgcttacggt?ttatggctaa?catctgtgat 720
atgtgagatt?tggtttgctg?tttcttgggt?gttggatcag?ttccctaaat?ggtctcctat 780
taaccgagaa?acttacatcg?accggttatc?cgcaagattc?gaaagagaag?gcgagcaatc 840
acagcttgca?gctgtagatt?tctttgttag?tacggtagat?ccattaaagg?agccaccttt 900
gataactgca?aacacggttc?tttcgatcct?cgcgcttgat?tatccggtgg?ataaagtctc 960
ttgctatgta?tctgatgatg?gtgctgcaat?gctttcgttt?gagtctttgg?ttgagacagc 1020
agattttgct?aggaaatggg?tacctttctg?caaaaagtac?tccatcgagc?cacgagctcc 1080
cgagttttac?ttctcgctta?aaatcgatta?cttgagggat?aaagttcaac?cttcttttgt 1140
gaaagaacgt?agagccatga?aaagagatta?tgaagagttt?aaaataagaa?tgaatgcttt 1200
agtcgccaag?gctcaaaaga?caccagaaga?aggatggaca?atgcaagatg?gaacatcttg 1260
gcccgggaac?aacactcgtg?accatcccgg?gatgattcag?gtttttcttg?gatatagcgg 1320
tgctcgcgac?attgaaggaa?atgaacttcc?aagattagtt?tacgtctcta?gagagaagag 1380
acctggttat?cagcatcaca?aaaaggccgg?ggcagagaac?gcattggtga?gggtgtctgc 1440
ggttttaacg?aatgctccat?tcattcttaa?ccttgattgt?gatcactacg?tcaacaatag 1500
caaagccgtg?cgtgaagcaa?tgtgcttttt?aatggatcct?gttgttggtc?aagacgtttg 1560
ctttgttcag?ttcccacaga?gatttgatgg?aatcgacaag?agtgatcgat?atgctaaccg 1620
caacattgtt?ttcttcgatg?ttaatatgag?agggcttgat?gggattcaag?gtccagttta 1680
tgttggtaca?ggtactgtct?ttagaagaca?agcactttac?ggatacagtc?caccttcaaa 1740
accgaggatt?ttaccgcaat?cttcatcatc?gtcgtgttgc?tgtctaacca?agaagaaaca 1800
acctcaagat?ccttccgaga?tttacaaaga?tgcaaagcga?gaagaacttg?atgctgcaat 1860
ctttaatctt?ggggacctag?acaactacga?tgagtacgac?agatcgatgc?tgatttcaca 1920
aacaagcttt?gagaaaacgt?ttggtctctc?tacggttttc?atcgagtcta?ctcttatgga 1980
gaatggcggt?gttcccgact?ctgtaaaccc?gtcaacgctc?atcaaagaag?ctattcatgt 2040
cattagctgt?ggatacgaag?agaaaactga?atggggaaaa?gaaataggat?ggatttacgg 2100
gtcgatcacc?gaagacattt?tgacgggttt?caagatgcat?tgtcgtggat?ggaggtcgat 2160
ttactgtatg?ccattaagac?cagcatttaa?aggatctgct?ccaatcaatc?tatcagatag 2220
gcttcaccag?gttctacgtt?gggctcttgg?ctcggttgag?atcttcctta?gccgacattg 2280
tcctttgtgg?tacggttgca?gcggaggccg?cctcaagttg?ctccagagat?tagcttatat 2340
aaacactatt?gtctacccat?tcacttcttt?gcctcttgtt?gcttactgta?ctcttccagc 2400
tatttgcctt?cttaccggca?aatttatcat?cccaacgcta?tcaaacctag?caagcatgct 2460
gtttctaggt?ctctttatat?caatcatctt?aacgagtgtc?ctcgagcttc?gatggagcgg 2520
agtcagtatc?gaagacttat?ggagaaacga?acagttttgg?gttattggag?gtgtctcagc 2580
tcatctcttt?gccgttttcc?aaggattcct?caaaatgctc?gctggtctcg?acacaaattt 2640
cacagtcaca?tcgaaaaccg?cagatgattt?agaattcggt?gagctttaca?ttgtcaaatg 2700
gacaactctc?ttgatccctc?cgacgtcact?tcttataatc?aacttggtcg?gagttgttgc 2760
tggattctct?gacgctctta?acaaaggtta?tgaagcttgg?ggacctttgt?ttgggaaggt 2820
atttttcgcc?ttttgggtga?ttcttcatct?ttatccattc?ctcaaaggtc?ttatgggaag 2880
acaaaacaga?acacctacta?ttgttattct?ctggtctatc?ttgcttgctt?ctgtgttttc 2940
acttgtttgg?gttcgtatca?atcctttcgt?ctccaaaacc?gatacgactt?ccctttctct 3000
gaactgtctt?ttgatcgatt?gctaagagaa?gatacgttat?gtttgtattt?tgaaagattg 3060
atcatatgtg?ttttggttgt?tttataaatt?ttcatatggt?tgcttgagcc?acaagttaag 3120
taatgttctt?attttagcaa?caagtcttgg?cgggttccgc?aagttaggtt?tctattgttt 3180
caacatcaac?atgtttttaa?aaagcaaaat?attttat 3217
<210>2
<211>985
<212>PRT
＜213〉Arabidopsis Arabidopis thaliana (Arabidopsis thaliana)
<400>2
Met?Met?Glu?Ser?Arg?Ser?Pro?Ile?Cys?Asn?Thr?Cys?Gly?Glu?Glu?Ile
1 5 10 15
Gly?Val?Lys?Ser?Asn?Gly?Glu?Phe?Phe?Val?Ala?Cys?His?Glu?Cys?Ser
20 25 30
Phe?Pro?Ile?Cys?Lys?Ala?Cys?Leu?Glu?Tyr?Glu?Phe?Lys?Glu?Gly?Arg
35 40 45
Arg?Ile?Cys?Leu?Arg?Cys?Gly?Asn?Pro?Tyr?Asp?Glu?Asn?Val?Phe?Asp
50 55 60
Asp?Val?Glu?Thr?Lys?Thr?Ser?Lys?Thr?Gln?Ser?Ile?Val?Pro?Thr?Gln
65 70 75 80
Thr?Asn?Asn?Thr?Ser?Gln?Asp?Ser?Gly?Ile?His?Ala?Arg?His?Ile?Ser
85 90 95
Thr?Val?Ser?Thr?Ile?Asp?Ser?Glu?Leu?Asn?Asp?Glu?Tyr?Gly?Asn?Pro
100 105 110
Ile?Trp?Lys?Asn?Arg?Val?Glu?Ser?Trp?Lys?Asp?Lys?Lys?Asp?Lys?Lys
115 120 125
Ser?Lys?Lys?Lys?Lys?Lys?Asp?Pro?Lys?Ala?Thr?Lys?Ala?Glu?Gln?His
130 135 140
Glu?Ala?Gln?Ile?Pro?Thr?Gln?Gln?His?Met?Glu?Asp?Thr?Pro?Pro?Asn
145 150 155 160
Thr?Glu?Ser?Gly?Ala?Thr?Asp?Val?Leu?Ser?Val?Val?Ile?Pro?Ile?Pro
165 170 175
Arg?Thr?Lys?Ile?Thr?Ser?Tyr?Arg?Ile?Val?Ile?Ile?Met?Arg?Leu?Ile
180 185 190
Ile?Leu?Ala?Leu?Phe?Phe?Asn?Tyr?Arg?Ile?Thr?His?Pro?Val?Asp?Ser
195 200 205
Ala?Tyr?Gly?Leu?Trp?Leu?Thr?Ser?Val?Ile?Cys?Glu?Ile?Trp?Phe?Ala
210 215 220
Val?Ser?Trp?Val?Leu?Asp?Gln?Phe?Pro?Lys?Trp?Ser?Pro?Ile?Asn?Arg
225 230 235 240
Glu?Thr?Tyr?Ile?Asp?Arg?Leu?Ser?Ala?Arg?Phe?Glu?Arg?Glu?Gly?Glu
245 250 255
Gln?Ser?Gln?Leu?Ala?Ala?Val?Asp?Phe?Phe?Val?Ser?Thr?Val?Asp?Pro
260 265 270
Leu?Lys?Glu?Pro?Pro?LeuIle?Thr?Ala?Asn?Thr?Val?Leu?Ser?Ile?Leu
275 280 285
Ala?Leu?Asp?Tyr?Pro?Val?Asp?Lys?Val?Ser?Cys?Tyr?Val?Ser?Asp?Asp
290 295 300
Gly?Ala?Ala?Met?Leu?Ser?Phe?Glu?Ser?Leu?Val?Glu?Thr?Ala?Asp?Phe
305 310 315 320
Ala?Arg?Lys?Trp?Val?Pro?Phe?Cys?Lys?Lys?Tyr?Ser?Ile?Glu?Pro?Arg
325 330 335
Ala?Pro?Glu?Phe?Tyr?Phe?Ser?Leu?Lys?Ile?Asp?Tyr?Leu?Arg?Asp?Lys
340 345 350
Val?Gln?Pro?Ser?Phe?Val?Lys?Glu?Arg?Arg?Ala?Met?Lys?Arg?Asp?Tyr
355 360 365
Glu?Glu?Phe?Lys?Ile?Arg?Met?Asn?Ala?Leu?Val?Ala?Lys?Ala?Gln?Lys
370 375 380
Thr?Pro?Glu?Glu?Gly?Trp?Thr?Met?Gln?Asp?Gly?Thr?Ser?Trp?Pro?Gly
385 390 395 400
Asn?Asn?Thr?Arg?Asp?His?Pro?Gly?Met?Ile?Gln?Val?Phe?Leu?Gly?Tyr
405 410 415
Ser?Gly?Ala?Arg?Asp?Ile?Glu?Gly?Asn?Glu?Leu?Pro?Arg?Leu?Val?Tyr
420 425 430
Val?Ser?Arg?Glu?Lys?Arg?Pro?Gly?Tyr?Gln?His?His?Lys?Lys?Ala?Gly
435 440 445
Ala?Glu?Asn?Ala?Leu?Val?Arg?Val?Ser?Ala?Val?Leu?Thr?Asn?Ala?Pro
450 455 460
Phe?Ile?Leu?Asn?Leu?Asp?Cys?Asp?His?Tyr?Val?Asn?Asn?Ser?Lys?Ala
465 470 475 480
Val?Arg?Glu?Ala?Met?Cys?Phe?Leu?Met?Asp?Pro?Val?Val?Gly?Gln?Asp
485 490 495
Val?Cys?Phe?Val?Gln?Phe?Pro?Gln?Arg?Phe?Asp?Gly?Ile?Asp?Lys?Ser
500 505 510
Asp?Arg?Tyr?Ala?Asn?Arg?Asn?Ile?Val?Phe?Phe?Asp?Val?Asn?Met?Arg
515 520 525
Gly?Leu?Asp?Gly?Ile?Gln?Gly?Pro?Val?Tyr?Val?Gly?Thr?Gly?Thr?Val
530 535 540
Phe?Arg?Arg?Gln?Ala?Leu?Tyr?Gly?Tyr?Ser?Pro?Pro?Ser?Lys?Pro?Arg
545 550 555 560
Ile?Leu?Pro?Gln?Ser?Ser?Ser?Ser?Ser?Cys?Cys?Cys?Leu?Thr?Lys?Lys
565 570 575
Lys?Gln?Pro?Gln?Asp?Pro?Ser?Glu?Ile?Tyr?Lys?Asp?Ala?Lys?Arg?Glu
580 585 590
Glu?Leu?Asp?Ala?Ala?Ile?Phe?Asn?Leu?Gly?Asp?Leu?Asp?Asn?Tyr?Asp
595 600 605
Glu?Tyr?Asp?Arg?Ser?Met?Leu?Ile?Ser?Gln?Thr?Ser?Phe?Glu?Lys?Thr
610 615 620
Phe?Gly?Leu?Ser?Thr?Val?Phe?Ile?Glu?Ser?Thr?Leu?Met?Glu?Asn?Gly
625 630 635 640
Gly?Val?Pro?Asp?Ser?Val?Asn?Pro?Ser?Thr?Leu?Ile?Lys?Glu?Ala?Ile
645 650 655
His?Val?Ile?Ser?Cys?Gly?Tyr?Glu?Glu?Lys?Thr?Glu?Trp?Gly?Lys?Glu
660 665 670
Ile?Gly?Trp?Ile?Tyr?Gly?Ser?Ile?Thr?Glu?Asp?Ile?Leu?Thr?Gly?Phe
675 680 685
Lys?Met?His?Cys?Arg?Gly?Trp?Arg?Ser?Ile?Tyr?Cys?Met?Pro?Leu?Arg
690 695 700
Pro?Ala?Phe?Lys?Gly?Ser?Ala?Pro?Ile?Asn?Leu?Ser?Asp?Arg?Leu?His
705 710 715 720
Gln?Val?Leu?Arg?Trp?Ala?Leu?Gly?Ser?Val?Glu?Ile?Phe?Leu?Ser?Arg
725 730 735
His?Cys?Pro?Leu?Trp?Tyr?Gly?Cys?Ser?Gly?Gly?Arg?Leu?Lys?Leu?Leu
740 745 750
Gln?Arg?Leu?Ala?Tyr?Ile?Asn?Thr?Ile?Val?Tyr?Pro?Phe?Thr?Ser?Leu
755 760 765
Pro?Leu?Val?Ala?Tyr?Cys?Thr?Leu?Pro?Ala?Ile?Cys?Leu?Leu?Thr?Gly
770 775 780
Lys?Phe?Ile?Ile?Pro?Thr?Leu?Ser?Asn?Leu?Ala?Ser?Met?Leu?Phe?Leu
785 790 795 800
Gly?Leu?Phe?Ile?Ser?Ile?Ile?Leu?Thr?Ser?Val?Leu?Glu?Leu?Arg?Trp
805 810 815
Ser?Gly?Val?Ser?Ile?Glu?Asp?Leu?Trp?Arg?Asn?Glu?Gln?Phe?Trp?Val
820 825 830
Ile?Gly?Gly?Val?Ser?Ala?His?Leu?Phe?Ala?Val?Phe?Gln?Gly?Phe?Leu
835 840 845
Lys?Met?Leu?Ala?Gly?Leu?Asp?Thr?Asn?Phe?Thr?Val?Thr?Ser?Lys?Thr
850 855 860
Ala?Asp?Asp?Leu?Glu?Phe?Gly?Glu?Leu?Tyr?Ile?Val?Lys?Trp?Thr?Thr
865 870 875 880
Leu?Leu?Ile?Pro?Pro?Thr?Ser?Leu?Leu?Ile?Ile?Asn?Leu?Val?Gly?Val
885 890 895
Val?Ala?Gly?Phe?Ser?Asp?Ala?Leu?Asn?Lys?Gly?Tyr?Glu?Ala?Trp?Gly
900 905 910
Pro?Leu?Phe?Gly?Lys?Val?Phe?Phe?Ala?Phe?Trp?Val?Ile?Leu?His?Leu
915 920 925
Tyr?Pro?Phe?Leu?Lys?Gly?Leu?Met?Gly?Arg?Gln?Asn?Arg?Thr?Pro?Thr
930 935 940
Ile?Val?Ile?Leu?Trp?Ser?Ile?Leu?Leu?Ala?Ser?Val?Phe?Ser?Leu?Val
945 950 955 960
Trp?Val?Arg?Ile?Asn?Pro?Phe?Val?Ser?Lys?Thr?Asp?Thr?Thr?Ser?Leu
965 970 975
Ser?Leu?Asn?Cys?Leu?Leu?Ile?Asp?Cys
980 985
Claims (10)
1, plant drought associated protein is the protein with one of following amino acid residue sequences:
1) the SEQ ID № in the sequence table: 2;
2) with SEQ ID № in the sequence table: 2 amino acid residue sequence is through replacement and/or disappearance and/or the interpolation and the protein relevant with plant drought of one or several amino-acid residue.
2, plant drought associated protein according to claim 1 is characterized in that: described plant drought associated protein has the SEQ ID № in the sequence table: 2 amino acid residue sequence.
3, the encoding gene of claim 1 or 2 described plant drought associated protein.
4, gene according to claim 3 is characterized in that: the cDNA gene of described plant drought associated protein has one of following nucleotide sequence:
1) SEQ ID № in the sequence table: 1 dna sequence dna;
2) SEQ ID № in the code sequence tabulation: the polynucleotide of 2 protein sequences;
3) under the rigorous condition of height can with SEQ ID № in the sequence table: the nucleotide sequence of the 1 dna sequence dna hybridization that limits;
4) with sequence table in SEQ ID №: 1 dna sequence dna that limits has 90% above homology, and the identical function protein DNA sequence of encoding.
5, the expression vector that contains claim 3 or 4 described plant drought associated protein encoding genes.
6, the clone that contains claim 3 or 4 described plant drought associated protein encoding genes.
7, the host bacterium that contains claim 3 or 4 described plant drought associated protein encoding genes.
8, the primer of amplification claim 3 or 4 described plant drought associated protein encoding genes.
9, a kind of method of utilizing the encoding gene enhancing plant drought resistance of claim 3 or 4 described plant drought resistance associated protein is the expression that suppresses the described plant drought associated protein encoding gene in the plant.
10, method according to claim 9, it is characterized in that: the expression method that suppresses the described plant drought resistance associated protein encoding gene in the plant comprises the method for plant viral vector mediated gene silencing, the gene silencing methods of the method for antisense technology silencer and siRNA mediation.
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| CNB2005100664361A CN1293095C (en) | 2005-04-25 | 2005-04-25 | Drought resistant correlative protein and coded gene of plant and application |
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Cited By (6)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN102203261A (en) * | 2008-06-13 | 2011-09-28 | 波夫曼斯种植公司 | Methods and means of increasing the water use efficiency of plants |
| CN111018959A (en) * | 2019-12-31 | 2020-04-17 | 中国农业大学 | Application of BMDR protein and coding gene thereof in regulating and controlling plant drought resistance |
| CN114644701A (en) * | 2020-12-21 | 2022-06-21 | 中国农业大学 | Use of proteins derived from corn and related biomaterials |
| CN114644696A (en) * | 2020-12-17 | 2022-06-21 | 中国农业大学 | Protein ZMPCPK 6 and coding gene and application thereof |
| CN114656537A (en) * | 2020-12-22 | 2022-06-24 | 中国农业大学 | GRMZM2G071330 protein and application thereof |
| CN114717243A (en) * | 2020-12-18 | 2022-07-08 | 中国农业大学 | GRMZM2G063882 gene, encoding protein, biological material and application thereof in plant drought resistance |
Family Cites Families (4)
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| US20040031072A1 (en) * | 1999-05-06 | 2004-02-12 | La Rosa Thomas J. | Soy nucleic acid molecules and other molecules associated with transcription plants and uses thereof for plant improvement |
| US7732667B2 (en) * | 2003-08-27 | 2010-06-08 | Syngenta Participations Ag | Transgenic plants and progeny and seed thereof |
| CN1333075C (en) * | 2003-09-18 | 2007-08-22 | 中国科学院遗传与发育生物学研究所 | Dry resisting related gene and its coding protein and application |
| CN1239704C (en) * | 2003-11-25 | 2006-02-01 | 中国农业大学 | Gene associated with plant salt resistance and drought resistance, encoded protein and application thereof |
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| Publication number | Priority date | Publication date | Assignee | Title |
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| CN102203261A (en) * | 2008-06-13 | 2011-09-28 | 波夫曼斯种植公司 | Methods and means of increasing the water use efficiency of plants |
| CN111018959A (en) * | 2019-12-31 | 2020-04-17 | 中国农业大学 | Application of BMDR protein and coding gene thereof in regulating and controlling plant drought resistance |
| CN111018959B (en) * | 2019-12-31 | 2021-06-25 | 中国农业大学 | Application of BMDR protein and coding gene thereof in regulating and controlling plant drought resistance |
| CN114644696A (en) * | 2020-12-17 | 2022-06-21 | 中国农业大学 | Protein ZMPCPK 6 and coding gene and application thereof |
| CN114644696B (en) * | 2020-12-17 | 2023-03-21 | 中国农业大学 | Protein ZMCPK6 and coding gene and application thereof |
| CN114717243A (en) * | 2020-12-18 | 2022-07-08 | 中国农业大学 | GRMZM2G063882 gene, encoding protein, biological material and application thereof in plant drought resistance |
| CN114644701A (en) * | 2020-12-21 | 2022-06-21 | 中国农业大学 | Use of proteins derived from corn and related biomaterials |
| CN114644701B (en) * | 2020-12-21 | 2023-03-21 | 中国农业大学 | Use of proteins derived from corn and related biomaterials |
| CN114656537A (en) * | 2020-12-22 | 2022-06-24 | 中国农业大学 | GRMZM2G071330 protein and application thereof |
| CN114656537B (en) * | 2020-12-22 | 2023-02-21 | 中国农业大学 | GRMZM2G071330 protein and application thereof |
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| CN1293095C (en) | 2007-01-03 |
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