CN101831436A - Method for breeding adverse-resistant plant - Google Patents
Method for breeding adverse-resistant plant Download PDFInfo
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- CN101831436A CN101831436A CN 201010116830 CN201010116830A CN101831436A CN 101831436 A CN101831436 A CN 101831436A CN 201010116830 CN201010116830 CN 201010116830 CN 201010116830 A CN201010116830 A CN 201010116830A CN 101831436 A CN101831436 A CN 101831436A
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Abstract
The invention discloses a method for breeding an adverse-resistant plant. The method provided by the invention for breeding the adverse-resistant plant comprises a step of guiding code gene of OsEIL1 protein into a start plant to obtain a transgenic plant of which the adverse resistance is higher than that of the start plant; the OsEIL1 protein is the protein of the following (a) or (b): (a) protein consisting of an amino acid sequence expressed as sequence 1 in a sequence table; and (b) protein consisting of the amino acid sequence of the sequence 1 by substituting and/or deleting and/or adding one or more amino acid residues, related with the adverse resistance of the plant and derived from the sequence 1. The code gene of the OsEIL1 protein is guided into the plant so that the adverse resistance (such as drought and low temperature stress) of the plant to abiotic stress can be obviously improved, and the plant can grow normally in the abiotic stress and is not stressed to die. The method provided by the invention can improve the adverse resistance of the plant to the abiotic stress and has broad application prospect.
Description
Technical field
The present invention relates to a kind of method of cultivating plant with adverse resistance.
Background technology
The shortage of water resources and the soil salinization are the severe facts that global agriculture production faces.Because drought is frequent, China's annual area suffered from drought reaches 3.27 hundred million mu, and along with global warming, drought will be on the rise, and is subjected to drought to threaten the farmland area may be above 700,000,000 mu.It is estimated that arid, low temperature and abiotic stress such as saline and alkaline reach 40% to the influence of crop yield, every year is caused hundred million kilograms of underproduction 700-800 because of lack of water in northern China main food producing region; Seasonal drought also frequently takes place in the abundant south of quantity of precipitation, as the annual area suffered from drought in Guangdong above 7,500,000 mu; Sichuan Province's crop area suffered from drought in 2006 reaches 2,100 ten thousand mu, accounts for 35% of sown area, has no harvest 3,970,000 mu, causes direct economic loss to reach 6,100,000,000 yuan.Paddy rice is as China's important crops, and water consumption accounts for 70% of agricultural water amount, so water resources shortage and arid, low temperature and abiotic stress such as saline and alkaline have become and influence the critical limitation of the rice high yield stable yields factor.Plant stress-resistance mechanism is very complicated, and improvement has certain effect though traditional breeding technique is for the resistance of crop, also has very big distance from people's desired destination.Cultivating anti-rice varieties against the current by biotechnology is to make full use of China saltings, alleviation shortage of water resources, the most economical and valid approach of assurance rice high yield stable yields, to ensureing that agricultural sustainable development has important practical significance.
Studies show that, after plant was subjected to arid, low temperature and abiotic stress such as saline and alkaline, the signal transduction by a series of complexity also activated specific transcriptional regulator, again by specific cis-acting elements, regulate and control the expression of a large amount of target genes, finally improve the resistance of reverse of plant.In improving the molecular breeding of crop to environment-stress, the transgenosis strategy of traditional single resistance functional gene has certain limitation to the improvement of farm crop resistance, and improvement or strengthen component in the signal transduction pathway of a key, just may regulate and control the expression of a plurality of downstreams resistance functional gene simultaneously, be more effective ways and the approach that improves crop anti-adversity.
Summary of the invention
The purpose of this invention is to provide a kind of method of cultivating plant with adverse resistance.
The method of cultivation plant with adverse resistance provided by the invention is that the proteic encoding gene of OsEIL1 is imported in the plant that sets out, and obtains the transgenic plant that resistance of reverse is higher than the described plant that sets out; Described OsEIL1 albumen is following (a) or protein (b):
(a) protein of forming by the aminoacid sequence shown in the sequence in the sequence table 1;
(b) with the aminoacid sequence of sequence 1 through the replacement of one or several amino-acid residue and/or disappearance and/or interpolation and relevant with plant stress tolerance by sequence 1 deutero-protein.
The proteic encoding gene of described OsEIL1 can be following 1) or 2) or 3) dna molecular:
1) dna molecular shown in the sequence 2 in the sequence table;
2) under stringent condition with 1) the dna sequence dna hybridization that limits and the dna molecular of coding stress tolerance correlative protein;
3) with 1) or 2) dna sequence dna that limits has 90% above homology, and the dna molecular of the stress tolerance correlative protein of encoding.
Above-mentioned stringent condition can be at 6 * SSC, in the solution of 0.5%SDS, 65 ℃ of hybridization down, uses 2 * SSC then, and 0.1%SDS and 1 * SSC, 0.1%SDS respectively wash film once.
Available existing plant expression vector construction contains the recombinant expression vector of described gene.
Described plant expression vector comprises the double base agrobacterium vector and can be used for the carrier etc. of plant micropellet bombardment.Described plant expression vector also can comprise 3 ' end untranslated zone of foreign gene, promptly comprises the dna fragmentation of polyadenylic acid signal and any other participation mRNA processing or genetic expression.The bootable polyadenylic acid of described polyadenylic acid signal joins 3 ' end of mRNA precursor, and the non-translational region of inducing (Ti) plasmid gene (as kermes synthetic enzyme Nos gene), plant gene (as soybean storage protein gene) 3 ' end to transcribe as the Agrobacterium crown-gall nodule all has similar functions.
When using described gene constructed recombinant plant expression vector, before its transcription initiation Nucleotide, can add any enhancement type promotor or constitutive promoter, as the ubiquitin promoter (Ubiquitin) of cauliflower mosaic virus (CAMV) 35S promoter, corn, they can use separately or be used in combination with other plant promoter; In addition, when using gene constructed plant expression vector of the present invention, also can use enhanser, comprise translational enhancer or transcriptional enhancer, these enhanser zones can be ATG initiator codon or neighboring region initiator codon etc., but must be identical with the reading frame of encoding sequence, to guarantee the correct translation of whole sequence.The source of described translation control signal and initiator codon is widely, can be natural, also can be synthetic.Translation initiation region can be from transcription initiation zone or structure gene.
For the ease of transgenic plant cells or plant being identified and screening, can process used plant expression vector, can produce the enzyme of colour-change or the gene of luminophor (gus gene, luciferase genes etc.) as adding the coding that in plant, to express, have the antibiotic marker thing (gentamicin marker, kantlex marker etc.) of resistance or anti-chemical reagent marker gene (as anti-weedkiller gene) etc.From the security consideration of transgenic plant, can not add any selected marker, directly with adverse circumstance screening transformed plant.
The proteic encoding gene of described OsEIL1 can import in the plant by pCAMBIA1301-OsEIL1; Described pCAMBIA1301-OsEIL1 inserts the recombinant plasmid that the proteic encoding gene of described OsEIL1 obtains in the multiple clone site of pCAMBIA 1301.Described pCAMBIA1301-OsEIL1 specifically can be and insert the recombinant plasmid that the proteic encoding gene of described OsEIL1 obtains between the BamH of pCAMBIA 1301 I and Pmac I restriction enzyme site.
Described plant with adverse resistance can be the plant of anti-abiotic stress.Described abiotic stress can be drought stress, low temperature stress (as 4 ℃) etc.
Utilize any carrier that can guide foreign gene in plant, to express,, can obtain anti-contrary ability enhanced transgenic cell line and transfer-gen plant the gene transfered plant cell of encoding said proteins.Carry that described expression carrier 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 the plant transformed tissue cultivating is become plant.By the plant transformed host both can be monocotyledons, it also can be dicotyledons, as nursery stock, flowers and plants etc., as cotton, soybean, rape, tobacco, Root or stem of Littleleaf Indianmulberry, Arabidopis thaliana, paddy rice (as the fine paddy rice of Japan), wheat, corn, cucumber, tomato, willow, turfgrass, lucerne place etc.
Described OsEIL1 albumen or its encoding gene can be applicable to cultivate plant with adverse resistance.
Make up OsEIL1 overexpression vector and rice transformation callus among the present invention, obtained the transgenic paddy rice strain system of a series of overexpressions.Cross and express OsEIL1 and can improve rice seedling significantly, make plant energy normal growth when running into abiotic stress such as arid and low temperature the arid and the patience of low temperature stress, unlikely coerce deadly.Method provided by the invention can improve the resistance of reverse of plant to abiotic stress, has broad application prospects.
Description of drawings
Fig. 1 is the structural representation of pCAMBIA1301-OsEIL1.
Fig. 2 is the expression level analytical results of OsEIL1 in the OsEIL1 overexpression plant.
Fig. 3 is the phenotype analytical before and after the OsEIL1 overexpression plant drought stress; A: the seedling of cultivating for two weeks; B: the seedling after arid is handled.
Fig. 4 is the phenotype analytical behind the OsEIL1 overexpression plant low temperature stress.
Embodiment
Following embodiment is convenient to understand better the present invention, but does not limit the present invention.Experimental technique among the following embodiment if no special instructions, is ordinary method.Used test materials among the following embodiment if no special instructions, is to buy from routine biochemistry reagent shop and obtains.% among the following embodiment if no special instructions, is the quality percentage composition.
" Japan is fine ": Institute of Crop Science, Chinese Academy of Agricultural Science, numbering WD-10576.
Acquisition of embodiment 1, OsEIL1 overexpression plant and resistance of reverse are identified
One, the structure of OsEIL1 overexpression vector pCAMBIA1301-OsEIL1
The a pair of Auele Specific Primer of nucleotide sequence (GENBANK ACCESSION NUMBER:DQ153245) design according to OsEIL1 is as follows:
Upstream primer: 5 '-C GG
ATGATGGGAGGTGGTCTGG-3 ';
Downstream primer: 5 '-TGCC
TCAGTAGTACCAATTCGAGC-3 '.
CDNA with japonica rice variety " Japan is fine " (Oryza sativa subsp.japonica cv.Nipponbare) is a template, carry out pcr amplification with above-mentioned Auele Specific Primer, reclaim pcr amplification product, carry out double digestion, reclaim enzyme and cut product with restriction enzyme BamHI and Pmac; With restriction enzyme BamH I and Pmac I double digestion pCAMBIA1301 (Cambia, GPO Box 3200, Canberra, ACT 2601, Australia), reclaim skeleton carrier; Described enzyme is cut product to be connected with described skeleton carrier, check order then, sequencing result shows, obtained pCAMBIA1301-OsEIL1 (, having inserted the OsEIL1 shown in the sequence 2 of sequence table between BamH I and the Pmac I restriction enzyme site) in the downstream of the 35S of pCAMBIA 1301 CaMV promotor.The structure iron of pCAMBIA1301-OsEIL1 is seen Fig. 1.
Two, the acquisition of OsEIL1 overexpression plant
1, with pCAMBIA1301-OsEIL1 by electric method for transformation import Agrobacterium LBA4404 (
Http:// www.cbs.knaw.nl/, NCCB bacteria/plasmids database, NCCB number2760), obtain the Agrobacterium of recombinating.
2, the acquisition of Agrobacterium tumefaciens mediated OsEIL1 overexpression paddy rice
Reorganization Agrobacterium bacterium liquid with step 1 is contaminated " Japan is fine " rice callus tissue; The callus of contaminating is blotted on aseptic filter paper, forward 24 ℃ of dark cultivations 2-4 days on the common substratum again to; The callus that cleans forwards on the selection substratum that contains Totomycin and carries out resistance screening; Kanamycin-resistant callus tissue after selecting forwards pre-differentiation culture medium culturing to and forwards division culture medium after 7-10 days again to and carry out illumination cultivation; Treat to forward to when seedling grows to 2-4cm growth 2-3 week in the test tube that root media is housed, well-grown seedling just can be transplanted to (T in the greenhouse after 2-3 days hardening
0Generation).Obtain the positive seedling (T1-8, T2-1, T1-6) of 3 strains.Respectively seedling is gone down to posterity, obtain T
1Generation.
Get the wild-type paddy rice (Japan is fine) and the T in 2 weeks respectively
1For rice seedling, extract mRNA and carry out reverse transcription, pass through the expression level (upstream primer: 5 '-AGCACGGAGAACAAGCCAT-3 ' of OsEIL1 in the fluorescent quantitative PCR technique analyzing rice behind the acquisition cDNA; Downstream primer: 5 '-GTTGAAACCAGAGCCGAACC-3 ').As 1, the expression level of OsEIL1 as shown in Figure 2 in T1-8, T2-1, three strains systems of T1-6 with the expression amount of OsEIL1 in the wild-type paddy rice.The result shows: T1-8, T2-1, T1-6 are OsEIL1 overexpression plant, and the expression amount of OsEIL1 is 2.5-3.8 a times of wild-type paddy rice, shows that the expression of this gene in transgenic paddy rice is significantly improved.
Three, OsEIL1 overexpression plant is to the patience of abiotic stress
T
1T is shown in representative
0The seed that produces for selfing and by plant that it grew up to.
Seed (T with the positive seedlings of 3 strains (T1-8, T2-1, T1-6)
1Generation) uses Totomycin to select to sprout, sprout after 3 days and to carry out following resistance functional analysis after seedling gone to soil; The seed of wild-type paddy rice (Japan is fine) is adopted same treatment, in contrast; Every kind of plant is got 30 plant, test triplicate, results averaged.
1, drought tolerance is identified
Paddy rice to 2 weeks of growing carries out arid processing (not watering in continuous 8 days), observes rice leaf then.Photo before and after rice drought is handled is seen Fig. 3.After arid 8 days, here the blade of wild-type paddy rice all withers; OsEIL1 overexpression paddy rice presents normal growth conditions, and blade is unfolded.The result shows that OsEIL1 has improved the drought tolerance of paddy rice.
2, change of the evaluation of OsEIL1 gene plant to low temperature patience
Paddy rice to 2 weeks of growing carries out subzero treatment (handling 2 days for 4 ℃), observes rice leaf.Photo after the paddy rice subzero treatment is seen Fig. 4.Handle after 2 days for 4 ℃, here withering appears in the blade of wild-type paddy rice; OsEIL1 overexpression paddy rice presents normal growth conditions.The result shows that OsEIL1 has improved paddy rice to cryogenic patience.
Sequence table
＜110〉Biological Technology institute, Chinese Academy of Agricultural Sciences
＜120〉a kind of method of cultivating plant with adverse resistance
<130>CGGNARY102120
<160>2
<210>1
<211>640
<212>PRT
＜213〉Japan fine (Oryza sativa subsp.japonica cv.Nipponbare)
<400>1
Met?Gly?Gly?Gly?Leu?Val?Met?Asp?Gln?Gly?Met?Met?Phe?Pro?Gly?Val
1 5 10 15
His?Asn?Phe?Val?Asp?Leu?Leu?Gln?Gln?Asn?Gly?Gly?Asp?Lys?Asn?Leu
20 25 30
Gly?Phe?Gly?Ala?Leu?Val?Pro?Gln?Thr?Ser?Ser?Gly?Glu?Gln?Cys?Val
35 40 45
Met?Gly?Glu?Gly?Asp?Leu?Val?Asp?Pro?Pro?Pro?Glu?Ser?Phe?Pro?Asp
50 55 60
Ala?Gly?Glu?Asp?Asp?Ser?Asp?Asp?Asp?Val?Glu?Asp?Ile?Glu?Glu?Leu
65 70 75 80
Glu?Arg?Arg?Met?Trp?Arg?Asp?Arg?Met?Lys?Leu?Lys?Arg?Leu?Lys?Glu
85 90 95
Leu?Gln?Leu?Ser?Arg?Gly?Lys?Asp?Pro?Ala?Gly?Gly?Val?Val?Gly?Asp
100 105 110
Pro?Ser?Lys?Pro?Arg?Gln?Ser?Gln?Glu?Gln?Ala?Arg?Arg?Lys?Lys?Met
115 120 125
Ser?Arg?Ala?Gln?Asp?Gly?Ile?Leu?Lys?Tyr?Met?Leu?Lys?Met?Met?Glu
130 135 140
Val?Cys?Arg?Ala?Gln?Gly?Phe?Val?Tyr?Gly?Ile?Ile?Pro?Glu?Lys?Gly
145 150 155 160
Lys?Pro?Val?Ser?Gly?Ala?Ser?Asp?Asn?Leu?Arg?Gly?Trp?Trp?Lys?Glu
165 170 175
Lys?Val?Arg?Phe?Asp?Arg?Asn?Gly?Pro?Ala?Ala?Ile?Ala?Lys?Tyr?Gln
180 185 190
Ala?Asp?Asn?Ala?Val?Pro?Gly?Phe?Glu?Ser?Glu?Leu?Ala?Ser?Gly?Thr
195 200 205
Gly?Ser?Pro?His?Ser?Leu?Gln?Glu?Leu?Gln?Asp?Thr?Thr?Leu?Gly?Ser
210 215 220
Leu?Leu?Ser?Ala?Leu?Met?Gln?His?Cys?Asp?Pro?Pro?Gln?Arg?Arg?Tyr
225 230 235 240
Pro?Leu?Glu?Lys?Gly?Val?Pro?Pro?Pro?Trp?Trp?Pro?Thr?Gly?Asp?Glu
245 250 255
Glu?Trp?Trp?Pro?Glu?Leu?Gly?Ile?Pro?Lys?Asp?Gln?Gly?Pro?Pro?Pro
260 265 270
Tyr?Lys?Lys?Pro?His?Asp?Leu?Lys?Lys?Ala?Trp?Lys?Val?Ser?Val?Leu
275 280 285
Thr?Ala?Val?Ile?Lys?His?Met?Ser?Pro?Asp?Ile?Glu?Lys?Ile?Arg?Arg
290 295 300
Leu?Val?Arg?Gln?Ser?Lys?Cys?Leu?Gln?Asp?Lys?Met?Thr?Ala?Lys?Glu
305 310 315 320
Ile?Ser?Thr?Trp?Leu?Ala?Val?Val?Lys?Gln?Glu?Glu?Glu?Leu?Tyr?Leu
325 330 335
Lys?Leu?Asn?Pro?Gly?Ala?Arg?Pro?Pro?Ala?Pro?Thr?Gly?Gly?Ile?Thr
340 345 350
Ser?Ala?Ile?Ser?Phe?Asn?Ala?Ser?Ser?Ser?Glu?Tyr?Asp?Val?Asp?Val
355 360 365
Val?Asp?Asp?Cys?Lys?Gly?Asp?Glu?Ala?Gly?Asn?Gln?Lys?Ala?Val?Val
370 375 380
Val?Ala?Asp?Pro?Thr?Ala?Phe?Asn?Leu?Gly?Ala?Ala?Met?Leu?Asn?Asp
385 390 395 400
Lys?Phe?Leu?Met?Pro?Ala?Ser?Met?Lys?Glu?Glu?Ala?Thr?Asp?Val?Glu
405 410 415
Phe?Ile?Gln?Lys?Arg?Ser?Ala?Ser?Gly?Ala?Glu?Pro?Glu?Leu?Met?Leu
420 425 430
Asn?Asn?Arg?Val?Tyr?Thr?Cys?His?Asn?Val?Gln?Cys?Pro?His?Ser?Asp
435 440 445
Tyr?Gly?Tyr?Gly?Phe?Leu?Asp?Arg?Asn?Ala?Arg?Asn?Ser?His?Gln?Tyr
450 455 460
Thr?Cys?Lys?Tyr?Asn?Asp?Pro?Leu?Gln?Gln?Ser?Thr?Glu?Asn?Lys?Pro
465 470 475 480
Ser?Pro?Pro?Ala?Ile?Phe?Pro?Ala?Thr?Tyr?Asn?Thr?Pro?Asn?Gln?Ala
485 490 495
Leu?Asn?Asn?Leu?Asp?Phe?Gly?Leu?Pro?Met?Asp?Gly?Gln?Arg?Ser?Ile
500 505 510
Thr?Glu?Leu?Met?Asn?Met?Tyr?Asp?Asn?Asn?Phe?Val?Ala?Asn?Lys?Asn
515 520 525
Leu?Ser?Asn?Asp?Asn?Ala?Thr?Ile?Met?Glu?Arg?Pro?Asn?Ala?Val?Asn
530 535 540
Pro?Arg?Ile?Gln?Ile?Glu?Glu?Gly?Phe?Phe?Gly?Gln?Gly?Ser?Gly?Ile
545 550 555 560
Gly?Gly?Ser?Asn?Gly?Gly?Val?Phe?Glu?Asp?Val?Asn?Gly?Met?Met?Gln
565 570 575
Gln?Pro?Gln?Gln?Thr?Thr?Pro?Ala?Gln?Gln?Gln?Phe?Phe?Ile?Arg?Asp
580 585 590
Asp?Thr?Pro?Phe?Gly?Asn?Gln?Met?Gly?Asp?Ile?Asn?Gly?Ala?Ser?Glu
595 600 605
Phe?Arg?Phe?Gly?Ser?Gly?Phe?Asn?Met?Ser?Gly?Ala?Val?Glu?Tyr?Pro
610 615 620
Gly?Ala?Met?Gln?Gly?Gln?Gln?Lys?Asn?Asp?Gly?Ser?Asn?Trp?Tyr?Tyr
625 630 635 640
<210>2
<211>1923
<212>DNA
＜213〉Japan fine (Oryza sativa subsp.japonica cv.Nipponbare)
<400>2
atgggaggtg?gtctggtgat?ggaccagggc?atgatgttcc?ccggcgtgca?caacttcgtg 60
gatctcctgc?agcagaacgg?cggcgacaag?aacctcggct?tcggcgcgct?cgtgccgcag 120
acgtcgtcgg?gggagcagtg?cgtgatgggg?gagggcgacc?tcgtggaccc?gccgccggag 180
agcttcccgg?acgccggtga?ggacgacagc?gacgacgacg?tggaggacat?cgaggagctg 240
gagcgccgca?tgtggcgcga?ccgcatgaag?ctgaagcggc?tcaaggagct?gcagctgagc 300
cggggcaagg?accccgcggg?cggcgtcgtg?ggcgacccgt?ccaagccgcg?gcagtcgcag 360
gagcaggcgc?ggcggaagaa?gatgtcgcgc?gcgcaggacg?gcatcctcaa?gtacatgctc 420
aagatgatgg?aggtgtgccg?cgcgcagggg?ttcgtgtacg?ggatcatccc?ggagaagggc 480
aagccggtga?gcggcgcctc?cgacaacctc?cgcggctggt?ggaaggagaa?ggtccgcttc 540
gaccgcaacg?gccccgccgc?catcgccaag?taccaggccg?acaacgccgt?cccgggcttc 600
gagagcgagc?tcgcctccgg?caccgggagc?ccgcactcgc?tgcaggagct?gcaggacacc 660
accctcgggt?cgctgctctc?ggcgctcatg?cagcactgcg?accctccgca?gcggcggtac 720
ccgctcgaga?agggcgtccc?tccgccgtgg?tggcccaccg?gcgacgagga?gtggtggccg 780
gagctcggca?tccccaagga?ccagggcccg?cctccgtaca?agaagcccca?tgacctcaag 840
aaggcctgga?aggtcagcgt?gctcaccgct?gtcatcaagc?acatgtcgcc?ggacatcgag 900
aagatccgcc?ggctggtccg?gcagtccaag?tgcctccagg?acaagatgac?cgccaaggag 960
atctccacct?ggctggccgt?cgtcaagcag?gaagaggagc?tgtacctgaa?gctgaacccc 1020
ggtgcccgcc?ctccggcacc?taccggcggc?atcaccagcg?ccatatcgtt?caacgccagc 1080
tcaagtgagt?acgacgtcga?cgtcgtcgac?gactgcaagg?gcgacgaggc?cggcaaccag 1140
aaggctgttg?ttgtcgccga?cccgaccgcg?ttcaacctcg?gcgcggctat?gctgaacgac 1200
aagttcctca?tgccggcgtc?catgaaggag?gaggccaccg?atgtcgagtt?catccagaag 1260
aggagcgcgt?ctggcgcgga?gcctgagctg?atgctgaaca?accgtgtcta?cacctgccac 1320
aatgtccagt?gcccgcatag?cgactatgga?tacgggttcc?ttgaccggaa?cgcgcgcaac 1380
agccaccaat?acacttgcaa?gtacaatgat?ccactccagc?agagcacgga?gaacaagcca 1440
tcgccaccgg?ccatcttccc?ggcaacctac?aacacgccga?accaggctct?gaacaatctg 1500
gatttcggcc?tgcccatgga?tggccagagg?tcaattacag?agctgatgaa?catgtacgac 1560
aacaacttcg?tggccaacaa?gaaccttagc?aacgacaatg?ccacgatcat?ggagaggcct 1620
aatgcagtca?acccaaggat?acagattgaa?gaaggctttt?ttggacaggg?aagtggcatc 1680
ggcggcagca?acggaggtgt?gttcgaagat?gtcaatggca?tgatgcagca?accgcagcag 1740
accaccccgg?cacagcagca?gttcttcatc?cgcgacgata?ctccattcgg?taaccagatg 1800
ggcgacatca?atggcgcatc?ggagttcagg?ttcggctctg?gtttcaacat?gtcaggtgcc 1860
gtcgaatacc?ccggcgcaat?gcagggccag?cagaagaatg?acggctcgaa?ttggtactac 1920
tga 1923
Claims (10)
1. a method of cultivating plant with adverse resistance is that the proteic encoding gene of OsEIL1 is imported in the plant that sets out, and obtains the transgenic plant that resistance of reverse is higher than the described plant that sets out; Described OsEIL1 albumen is following (a) or protein (b):
(a) protein of forming by the aminoacid sequence shown in the sequence in the sequence table 1;
(b) with the aminoacid sequence of sequence 1 through the replacement of one or several amino-acid residue and/or disappearance and/or interpolation and relevant with plant stress tolerance by sequence 1 deutero-protein.
2. the method for claim 1, it is characterized in that: the proteic encoding gene of described OsEIL1 is following 1) or 2) or 3) dna molecular:
1) dna molecular shown in the sequence 2 in the sequence table;
2) under stringent condition with 1) the dna sequence dna hybridization that limits and the dna molecular of coding stress tolerance correlative protein;
3) with 1) or 2) dna sequence dna that limits has 90% above homology, and the dna molecular of the stress tolerance correlative protein of encoding.
3. method as claimed in claim 2 is characterized in that: the proteic encoding gene of described OsEIL1 imports in the plant by pCAMBIA1301-OsEIL1; Described pCAMBIA1301-OsEIL1 inserts the recombinant plasmid that the proteic encoding gene of described OsEIL1 obtains in the multiple clone site of pCAMBIA 1301.
4. method as claimed in claim 3 is characterized in that: described pCAMBIA1301-OsEIL1 inserts the recombinant plasmid that the proteic encoding gene of described OsEIL1 obtains between the BamH of pCAMBIA1301 I and Pmac I restriction enzyme site.
5. as arbitrary described method in the claim 1 to 4, it is characterized in that: described plant with adverse resistance is the plant of anti-abiotic stress.
6. method as claimed in claim 5 is characterized in that: described abiotic stress is drought stress or low temperature stress.
7. method as claimed in claim 6 is characterized in that: described low temperature is 4 ℃.
8. as arbitrary described method in the claim 1 to 7, it is characterized in that: the described plant that sets out is dicotyledons or monocotyledons.
9. method as claimed in claim 8 is characterized in that: the described plant that sets out is cotton, soybean, rape, corn, paddy rice or tobacco, preferred Japanese fine paddy rice.
10.OsEIL1 albumen or its encoding gene application in cultivating plant with adverse resistance.
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Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102337291A (en) * | 2011-07-27 | 2012-02-01 | 湖南农业大学 | Method for gene transient expression of citrus |
| CN105028173A (en) * | 2015-05-20 | 2015-11-11 | 阮积恩 | Hybridization breeding method of high-phosphorus-efficiency maize |
| CN111793119A (en) * | 2019-04-04 | 2020-10-20 | 中国科学院遗传与发育生物学研究所 | Protein for regulating and controlling plant drought resistance, coding gene and application thereof |
| CN113527451A (en) * | 2020-04-21 | 2021-10-22 | 中国农业科学院作物科学研究所 | Wheat heat stress related protein TaANK and coding gene and application thereof |
| CN116410279A (en) * | 2021-12-31 | 2023-07-11 | 中国科学院植物研究所 | Protein related to regulation of rice abiotic stress tolerance, related biological material and application thereof |
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Cited By (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102337291A (en) * | 2011-07-27 | 2012-02-01 | 湖南农业大学 | Method for gene transient expression of citrus |
| CN105028173A (en) * | 2015-05-20 | 2015-11-11 | 阮积恩 | Hybridization breeding method of high-phosphorus-efficiency maize |
| CN111793119A (en) * | 2019-04-04 | 2020-10-20 | 中国科学院遗传与发育生物学研究所 | Protein for regulating and controlling plant drought resistance, coding gene and application thereof |
| CN113527451A (en) * | 2020-04-21 | 2021-10-22 | 中国农业科学院作物科学研究所 | Wheat heat stress related protein TaANK and coding gene and application thereof |
| CN113527451B (en) * | 2020-04-21 | 2023-07-14 | 中国农业科学院作物科学研究所 | Wheat heat stress related protein TaANK, and coding gene and application thereof |
| CN116410279A (en) * | 2021-12-31 | 2023-07-11 | 中国科学院植物研究所 | Protein related to regulation of rice abiotic stress tolerance, related biological material and application thereof |
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