CN110129291A - Barley moisture-proof controlling gene HvACO1, albumen and its application in breeding - Google Patents
Barley moisture-proof controlling gene HvACO1, albumen and its application in breeding Download PDFInfo
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- CN110129291A CN110129291A CN201910302541.2A CN201910302541A CN110129291A CN 110129291 A CN110129291 A CN 110129291A CN 201910302541 A CN201910302541 A CN 201910302541A CN 110129291 A CN110129291 A CN 110129291A
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Abstract
The invention discloses barley moisture-proof controlling gene HvACO1, albumen and its applications in breeding.Barley moisture-proof geneHvACO1CDS sequence as shown in SEQ ID NO:1.Clone obtains moisture-proof gene to the present invention from moisture-proof barley for the first timeHvACO1, and prove that the gene is related with barley moisture-proof, it, will using the method for genetic engineeringHvACO1Gene overexpression is into arabidopsis, and compared with the control group, transgenic plant moisture-proof ability significantly increases, explanationHvACO1The overexpression of gene improves the moisture-proof of plant.In the present inventionHvACO1The technologies such as the clone of gene and conversion, transgenosis are that the molecule mechanism for studying barley moisture-proof and Breeding Application are laid a good foundation, and have wide Breeding Application prospect and certain economic value.
Description
Technical field
The invention belongs to gene engineering technology field more particularly to a barley moisture-proof controlling genesHvACO1And its breeding
Using.
Background technique
To harm caused by crop normal growth and development when wet injury is soil moisture saturation or supersaturation.Wet injury is in the world
There is generation in range, wet injury occurs often in the states such as North America, Australia, China, India especially.According to the United Nations's grain and agricultural
The data that tissue is announced show that there are about 10% cultivated areas to be influenced by wet injury in world wide.Barley Dry crop, wet injury
Seriously affect barley production and quality.Wet injury can generally make barley underproduction 20%-30%, serious even to have no harvest.
Wet injury stress hinders the free flow of gas in plant rhizosphere tissue, and the photosynthesis and breathing for influencing plant are made
With.Plant can regulate and control Translation of Programmed Cell Death in Plants and newborn adventitious root and aerating tissue by bio-hormones such as synthesizing ethylenes
Formation, adapt to the adverse circumstance environment of anoxic, the moisture-proof that gene pairs relevant to ethylene synthase improves plant has adjustment effect.
Ethylene synthetase (ACS) and ethylene enzyme (ACO) are two key enzymes in ethylene synthase approach, and existing research shows will
CottonGhACOGene is transferred in arabidopsis,GhACOGene expression enhances the biosynthesis of ethylene in arabidopsis body, in turn
Arabidopsis is enhanced to the tolerance of abiotic stress.In recent years, about the anti contravariance related genes such as crop moisture-proof, salt tolerant, drought-enduring
Clone and the existing many report of breeding utilization, provide the new technological approaches of one kind for the breeding of resistance to inverse crop varieties.Currently,
The research of the relevant gene of barley moisture-proof is concentrated mainly on QTL positioning level, there is not yet barley moisture-proof gene cloning is answered with breeding
It is reported with research.
Summary of the invention
It is an object of the present invention to provide a barley moisture-proof controlling genesHvACO1And its Breeding Application, it clones and constructsHvACO1Expression vector is imported in arabidopsis using transgenic technology, carries out wet injury Stress treatment to transgenic arabidopsis,
VerifyingHvACO1Moisture-proof function, provide genetic resources and theoretical foundation for barley moisture-proof breed improvement.
In order to achieve the above-mentioned object of the invention, the technical scheme adopted by the invention is as follows: a kind of barley moisture-proof regulates and controls related egg
White, the albumen is for following (a) or (b):
(a) protein that the amino acid sequence shown in SEQ ID NO:2 forms；Particular sequence is MEIPVIDLQGLDGDASQ
RSQTMARLHEACKDWGFFWVDSHGVDAALMEEVKRFVYAHYDEHLKDRFYASDLAKDLLLPAEESKAVSGEVDWET
AYFIRHRPANNVADFPEIPPATREMLDVYIGQMVSLAERLAECMSLNLGLDGGRVKDTFAPPFVGTKFAMYPACPR
PDLLWGLRAHTDAGGIILLLQDDVVGGLEFFRGDREWVPVGPTKGSRIFVNLGDQLEVMSGGAYRSVLHRVAAVAE
GRRLSVATFYNPGAEAVVAPAPTARQPAAQVYPGPYRFGDYLDYYQGTKFADKAARLQAVKELFGSRILPD；
(b) by the amino acid sequence of SEQ ID NO:2 by one or several amino acid residues substitution and/or missing and/or
Addition and the protein as derived from SEQ ID NO:2 relevant to the regulation of barley moisture-proof.
The present invention also provides the genes of coding foregoing proteins.The gene is DNA any in following (a1)-(a3)
Molecule；
(a1) DNA molecular shown in SEQ ID NO:1；
(a2) hybridize under strict conditions with (a1) DNA sequence dna limited and the DNA of encoding barley moisture-proof regulation GAP-associated protein GAP divides
Son；
(a3) at least have 70% with (a1) DNA sequence dna limited, at least have 75%, at least with 80%, at least with 85%, extremely
Less with 90%, at least with 95%, at least with 96%, at least with 97%, at least with 98% or at least have it is 99% homologous
Property and encoding barley moisture-proof regulation GAP-associated protein GAP DNA molecular.
The present invention also provides the expression cassette containing forementioned gene, recombinant vector, recombinant microorganism or transgenic cell lines.
It is a further object to provide the applications of (b1) or (b2) or (b3):
(b1) albumen, or, the gene, or, containing the expression cassette of the gene, recombinant vector, recombinant microorganism or turning
Gene cell system, the application in anti-barley wet injury stress；
(b2) albumen, or, the gene, or, containing the expression cassette of the gene, recombinant vector, recombinant microorganism or turning
Gene cell system, the application in cultivation barley, arabidopsis moisture-proof new varieties；
(b3) albumen, or, the gene, or, containing the expression cassette of the gene, recombinant vector, recombinant microorganism or turning
Gene cell system, the application in anti-arabidopsis wet injury stress.
The present invention also provides a kind of methods for cultivating plant moisture-proof kind, and the gene is transferred to purpose plant or institute
Expression cassette, the recombinant vector stated convert purpose plant, obtain genetically modified plants；The wet injury stress resistance of the genetically modified plants is high
In the purpose plant.The purpose plant is barley, arabidopsis.The resistance to wetting phase of barley described in the Expressed in Transgenic Plant
Close albumen.
HvACO1DNA segment described in gene is as shown in sequence table SEQ ID NO.1, or substantially corresponds to SEQ ID
DNA sequence or its function shown in NO.1 are equivalent to the Partial Fragment of sequence shown in SEQ ID NO.1.The gene is carried out
Sequence analysis showsHvACO1Gene coding region overall length be 952 bp, coding 317 amino acid, molecular weight 35.02kDa, etc.
Electricity point is 5.16, contains conservative D10x-N and 20G-FelI-Oxy structural domain.Overexpress sequence shown in sequence table SEQ ID NO.1
Column can enhance the plants such as arabidopsis and barley to the patience of wet injury.
Said gene can be applied to improvement barley moisture-proof ability, and concrete operations are as follows:
(1) acquisition of gene: proteomic assays discovery is carried out to the root system of the barley TF58 of resistance to wet stock, wet injury stress causes
In root systemHvACO1The obvious up-regulated expression of gene extracts TF58 root system total serum IgE, is arrived using RT-PCR amplificationHvACO1Gene sequence
Column, amplified production is connected on pGEM-Teasy carrier, obtains target gene clone through sequencing.
(2) building of expression vector and genetic transformation: barley is constructed using the Gateway technology of Invitrogen companyHvACO1Gene overexpression vector.Using BP Clonase enzyme, target gene is connect with pDONR221 carrier, structure
Entry vector is built, LR Clonase enzyme is recycled, entry vector is connect with pB2GW7 carrier, is constructedHvACO1Base
Because of overexpression vector.It will by frozen-thawed methodHvACO1Gene overexpression vector imports in Agrobacterium Gv3101.Utilize agriculture bar
The genetic transformation that bacterium mediates, willHvACO1It is expressed in channel genes arabidopsis.Sun is screened by antibiotic-screening, RT-PCR etc.
Property transgenic Arabidopsis plants.
(3) transgenic plant moisture-proof identification andHvACO1Gene function verifying: by 35 days transgenosis of normal growth and
WT strain carries out submerging treatment.After submerging treatment 2 weeks, observe that the moisture-proof of transgenic plant is considerably better than wild type plant
The phenotype of strain, to proveHvACO1Gene is remarkably improved the moisture-proof of arabidopsis.
The present invention provides a kind of new method to the resistance of wet injury for improvement plant.It is cultivated by genetic engineering means resistance to
Wet plant overcomes the shortcomings of traditional breeding method, and not only breeding cycle is short, and it is easy to operate, be easy to get highly resistance material.The present invention
It is cloned into barley moisture-proof gene for the first timeHvACO1, and the gene is transferred in arabidopsis by mediated by agriculture bacillus method, through overly moist
Evil identification and analysis, it was demonstrated that transgenic plant is significantly improved compared with the moisture-proof ability of WT lines, it was demonstrated that gene of the invention exists
It is with a wide range of applications in terms of improving barley moisture-proof.
Detailed description of the invention
Fig. 1 barley HvACO1 gene order RT-PCR amplified production,
Marker:DL2000DNA Marker (the precious biology in Dalian), by 2,000bp, 1,000bp, 750bp, 500bp, 250bp
And six DNA segment compositions of 100bp；
Fig. 2 turnsHvACO1Gene arabidopsis strain and WT strain RT-PCR are tested and analyzed；
Fig. 3 arabidopsis transgenic line and the sex expression of WT strain moisture-proof and the variation of some growth index,
(A) phenotype of transgenic arabidopsis and wild type under normal growing conditions and after waterflooding 2 weeks；(B) plant height；(C) stem
Leaf fresh weight；(D) cauline leaf dry weight；(E) root length；(F) survival rate.
Specific embodiment
Invention is further described in detail combined with specific embodiments below.
Embodiment 1HvACO1The clone of gene
Proteome analysis is carried out using barley strain TF58 as a result, designHvACO1The special primer P of gene1Forward primer:
5 '-ATGGACATGGAGATCCCGGT-3 ' (SEQ ID NO.3) and P2Reverse primer: 5 '-TCAATCGGGCAGAATCCGTG-
3 ' (SEQ ID NO.4) extract moisture-proof barley strain TF58 root system total serum IgE using CTAB method, and reverse transcription is at cDNA, then with
CDNA is template, utilizes primer P1And P2Amplify the CDS sequence of the barley moisture-proof gene as shown in SEQ ID NO:1, geneHvACO1CDS sequence 952bp (Fig. 1).
Specific step is as follows:
(1) CTAB (cetyl trimethylammonium bromide) Extraction buffer [2% (W/V) CTAB, NaCl is added into centrifuge tube
1 .4mol/L, EDTA (ethylenediamine tetra-acetic acid) 20mmol/L, TrisHCl 100mmol/L, 2% (W/V) PVP] and 10% β-
Mercaptoethanol preheats in water-bath；
(2) grind barley root system liquid nitrogen is cooling, be added in extracting solution, mix, 65 DEG C water-bath 10 minutes；
(3) isometric chloroform is added: isoamyl alcohol (volume ratio 24:1) mixed liquor is mixed by inversion, and stands 10min, and 4 DEG C
12000g is centrifuged 10min；
(4) supernatant is taken, is repeated step (3)；
(5) supernatant is taken, the LiCl of final concentration of 2mol/L, ice bath 10-12 hours, 11000rpm, 4 DEG C centrifugation 15min is added,
Supernatant is abandoned, cleans precipitating twice with 75% ethyl alcohol, is dissolved in stand-by in suitable DEPC (pyrocarbonic acid diethyl ester) processing water；
(6) it is template that root system total serum IgE is extracted from barley variety TF58, (is purchased from Thermo Fisher using reverse transcriptase
Scientific company) by its reverse transcription synthesize first chain of cDNA, reaction condition are as follows: 65 DEG C of 5min, 42 DEG C of 50min, 70 DEG C
10min；
(7) barley ethylene enzyme gene is amplified from the cDNA that RNA reverse transcription obtains using above-mentioned primer P1 and P2HvACO1CDS sequence；
Reaction condition: 94 DEG C of initial denaturation 4min；94 DEG C of 30sec, 55 DEG C of 30sec, 72 DEG C of 1min, 33 circulations；72 DEG C of extensions
10min.The PCR product that amplification obtains is connected into pMD18-T carrier (purchased from precious bioengineering Dalian Co., Ltd), converts large intestine
Bacillus competent cell, screening positive clone are simultaneously sequenced, and obtain required full-length gene.It extracts and carries from positive colonyHvACO1The plasmid of gene C DS sequence.
Embodiment 2HvACO1The building of gene overexpression carrier and genetic transformation
In order to preferably analyze geneHvACO1Biological function, which is overexpressed in arabidopsis, utilize
The Gateway technology of Invitrogen company constructs barleyHvACO1Gene overexpression vector.Utilize BP Clonase
Target gene is connect by enzyme with pDONR221 carrier, constructs entry vector, then utilizes LR Clonase enzyme,
Entry vector is connect with pB2GW7 carrier, constructs the overexpression vector containing target gene.It will be contained by frozen-thawed methodHvACO1The overexpression vector of gene imports in Agrobacterium Gv3101.It, will using Agrobacterium-mediated genetic transformation methodHvACO1
Coded sequence is imported in the plants such as arabidopsis and is expressed.Positive transgenic plant is screened by antibiotic-screening and RT-PCR etc..
It is as follows that this tests practical genetic transforming method:
(1) culture of Agrobacterium
With corresponding resistance selection solid LB media (10g/L peptone, 5g/L yeast extract, 10g/L sodium chloride,
Kan 100mg/L, agar 1.5g/L) on preculture carry target geneHvACO1Agrobacterium 48 hours, cultivation temperature 28
℃；Picking preculture Agrobacterium single colonie is inoculated in LB liquid medium (the 10g/L peptone, 5g/L ferment of corresponding resistance selection
Female extract, 10g/L sodium chloride, Kan 100mg/L) in, it is stayed overnight in 28 DEG C of 200rpm shaking table cultures, until bacterial concentration OD600
Value is about the .0 of 0 .8~1.
(2) inflorescence method infects arabidopsis
Using the inflorescence dip method arabidopsis thaliana transformation of improvement.Picking contains the Agrobacterium single colonie of target gene plasmid, is transferred to
In 100mL LB liquid medium, it is placed in 200r/min in 28 DEG C of shaking tables and is incubated overnight, Agrobacterium bacterial concentration is made to reach OD600
In 0 .8~1 .0,4000r/min is centrifuged 10min, after abandoning supernatant, contains 50 μ L of sucrose 5g, SilwetL-77 with 100mL
LB liquid medium suspension Agrobacterium bacterium solution again.By the back-off of arabidopsis flowerpot in the beaker containing bacteria suspension, by quasi- south
The inflorescence of mustard is immersed in 10~20s in bacterium solution and is placed in 25 DEG C of temperature, humidity 60%, intensity of illumination after the lower culture for 24 hours of dark
It is cultivated in 3000~4000lx and the illumination box of 16h/8h photoperiod, waters and cultivate plant every other day, harvested to Post flowering
Seed.After disinfection 5min in the arabidopsis seed of harvest 8%NaClO alcoholic solution (95% alcohol), trained in selection
It supports and is screened on base (MS+50mg/L kanamycins+7g/L agar+30g/L sucrose).
Embodiment 3HvACO1Gene transgenic T3 is detected for seedling RT-PCR
In order to verify transgenic arabidopsis T3 for strain moisture-proof ability change whether be transferred toHvACO1Gene is related, uses
RT-PCR method is in partial transgenic Arabidopsis plantHvACO1Gene expression is detected, and as a result sees Fig. 2, it is known that turns base
Because arabidopsis T3 for strain moisture-proof ability change be transferred toHvACO1The overexpression of gene is related.
Specific step is as follows:
It is extracted from transgenic arabidopsis T3 3 strains of generation using TRIZOL reagent (purchased from precious bioengineering Dalian Co., Ltd)
The total serum IgE (extracting method is operated referring to TRIZOL reagent specification) of plant, (is purchased from Thermo Fisher using reverse transcriptase
Scientific company) by its reverse transcription synthesize first chain of cDNA, reaction condition be 65 DEG C of 5min, 42 DEG C of 50min, 70 DEG C
10min.Detection first is carried out to the cDNA that reverse transcription obtains with the reference gene Actin of report and concentration adjusts, carries out PCR inspection
It surveys, it is ensured that reference gene can expand in control wildtype Arabidopsis thaliana and transgenic Arabidopsis plants.Then, according to
The sequence of HvACO1 gene carries out RT-PCR detection, reaction condition are as follows: 94 DEG C of initial denaturation 4min using primer P1 and P2；94℃
30sec, 55 DEG C of 30sec, 72 DEG C of 1 .5min, 33 circulations；72 DEG C of extension 10min.The agarose gel electrophoresis knot of amplified production
Fruit shows (Fig. 2), and the expression of HvACO1 gene is detected in 3 transgenic lines.
4 turns of embodimentHvACO1The measurement of Arabidopsis plant moisture-proof
It is obtained from screeningHvACO1Genetic transformation arabidopsis T3 for selecting each 3 strain kinds at random in positive transgenic strain
Son compares (WT) with wildtype Arabidopsis thaliana seed, and culture is to 4 leaf phases on 1/2MS culture medium, by seedling replanting to equipped with nutrition
It is middle in the controlled environment chamber to cultivate 35 days in the basin alms bowl of soil.Transgenic line and wild type are put into water tank, carried out at waterflooding
Reason after waterflooding 2 weeks, observes the difference of transgenic line and wild type material moisture-proof, and measures the height of seedling of different materials, Ye Lv
The characters such as cellulose content, plant fresh weight, dry weight, root length, survival rate.Wild type plant height has dropped 49.09%, and 3 transgenosis
Strain has dropped 11.70%, 11.40%, 10.28% respectively；Wild type chlorophyll content has dropped 61.56%, and 3 transgenic lines
System has dropped 20.45%, 31.82%, 34.23% respectively；Wild type cauline leaf fresh weight has dropped 65.8%, 3 transgenic line difference
Have dropped 36.11%, 42.25%, 44.03%；Wild type cauline leaf dry weight has dropped 51.01%, and 3 transgenic lines have dropped respectively
17.98%, 36.48%, 31.10%；Wild type root long has dropped 74.87%, and 3 transgenic lines have dropped 49.62% respectively,
40.59%, 38.39%；The survival rate of wild type material is only 27.61%, and the survival rate of transgenic line is substantially higher Yu Feizhuan
Genetic material, respectively 68.85%, 68.79% and 71.82%.The result shows that passing through overexpression in arabidopsisHvACO1Gene
Transgenic line is clearly enhanced to the resistance of wet injury.
Basic principles and main features and advantages of the present invention of the invention have been shown and described above.The skill of the industry
Art personnel it should be appreciated that the present invention is not limited to the above embodiments, the above embodiments and description only describe
The principle of the present invention, without departing from the spirit and scope of the present invention, various changes and improvements may be made to the invention, these
Changes and improvements all fall within the protetion scope of the claimed invention.The claimed scope of the invention by appended claims and
Its equivalent thereof.
Sequence table
<110>Yangzhou University
<120>barley moisture-proof controlling gene HvACO1, albumen and its application in breeding
<130> xhx2019041601
<141> 2019-04-16
<160> 4
<170> SIPOSequenceListing 1.0
<210> 1
<211> 952
<212> DNA
<213> Hordeum vulgare L.
<400> 1
atggagatcc cggtcatcga cctccagggg ctcgacggcg acgcctccca gcggtcgcaa 60
accatggcgc ggctccacga agcctgcaag gactggggct tcttctgggt ggacagccat 120
ggcgtcgacg ccgcgctgat ggaggaggtg aagcgcttcg tgtatgccca ctacgacgag 180
catctcaagg ataggttcta cgcctcggac ctcgccaaag acctgctgct gccggcggag 240
gaatccaaag ccgtctccgg tgaggtagac tgggagaccg cctacttcat ccggcaccgt 300
cccgccaaca acgtcgccga cttcccggag atcccgccgg ccacacgaga gatgctcgac 360
gtgtacatcg gacagatggt gtcgctcgca gagcggctgg ccgagtgcat gagcctgaac 420
ctgggcctgg acggcggccg cgtcaaagac accttcgcgc cgccgttcgt cgggaccaag 480
ttcgccatgt acccggcctg cccgcggccg gacctcctgt ggggcctccg cgcccacacc 540
gacgccggcg gcatcatcct gctcctgcag gacgacgtgg tcggcgggct ggagttcttc 600
aggggcgacc gggagtgggt ccccgtgggc cccaccaagg gcagcagaat cttcgtcaac 660
ctcggggacc agctggaggt gatgagcggc ggcgcctaca ggagcgtgct gcaccgcgtc 720
gccgccgtcg cggaggggcg gcggctgtcg gtggcgacgt tctacaaccc cggggcggaa 780
gccgtcgtgg cgccggcgcc cacggcgagg cagccggcgg cgcaggtgta ccccgggccg 840
tacaggttcg gcgactacct ggactactac cagggcacca agtttgccga caaggcggcg 900
aggttgcagg ccgtcaagga gctgttcggc tcacggattc tgcccgatat ga 952
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Met Glu Ile Pro Val Ile Asp Leu Gln Gly Leu Asp Gly Asp Ala Ser
1 5 10 15
Gln Arg Ser Gln Thr Met Ala Arg Leu His Glu Ala Cys Lys Asp Trp
20 25 30
Gly Phe Phe Trp Val Asp Ser His Gly Val Asp Ala Ala Leu Met Glu
35 40 45
Glu Val Lys Arg Phe Val Tyr Ala His Tyr Asp Glu His Leu Lys Asp
50 55 60
Arg Phe Tyr Ala Ser Asp Leu Ala Lys Asp Leu Leu Leu Pro Ala Glu
65 70 75 80
Glu Ser Lys Ala Val Ser Gly Glu Val Asp Trp Glu Thr Ala Tyr Phe
85 90 95
Ile Arg His Arg Pro Ala Asn Asn Val Ala Asp Phe Pro Glu Ile Pro
100 105 110
Pro Ala Thr Arg Glu Met Leu Asp Val Tyr Ile Gly Gln Met Val Ser
115 120 125
Leu Ala Glu Arg Leu Ala Glu Cys Met Ser Leu Asn Leu Gly Leu Asp
130 135 140
Gly Gly Arg Val Lys Asp Thr Phe Ala Pro Pro Phe Val Gly Thr Lys
145 150 155 160
Phe Ala Met Tyr Pro Ala Cys Pro Arg Pro Asp Leu Leu Trp Gly Leu
165 170 175
Arg Ala His Thr Asp Ala Gly Gly Ile Ile Leu Leu Leu Gln Asp Asp
180 185 190
Val Val Gly Gly Leu Glu Phe Phe Arg Gly Asp Arg Glu Trp Val Pro
195 200 205
Val Gly Pro Thr Lys Gly Ser Arg Ile Phe Val Asn Leu Gly Asp Gln
210 215 220
Leu Glu Val Met Ser Gly Gly Ala Tyr Arg Ser Val Leu His Arg Val
225 230 235 240
Ala Ala Val Ala Glu Gly Arg Arg Leu Ser Val Ala Thr Phe Tyr Asn
245 250 255
Pro Gly Ala Glu Ala Val Val Ala Pro Ala Pro Thr Ala Arg Gln Pro
260 265 270
Ala Ala Gln Val Tyr Pro Gly Pro Tyr Arg Phe Gly Asp Tyr Leu Asp
275 280 285
Tyr Tyr Gln Gly Thr Lys Phe Ala Asp Lys Ala Ala Arg Leu Gln Ala
290 295 300
Val Lys Glu Leu Phe Gly Ser Arg Ile Leu Pro Asp
305 310 315
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Claims (8)
1. a kind of barley moisture-proof regulates and controls GAP-associated protein GAP, which is characterized in that the albumen is for following (a) or (b):
(a) protein that the amino acid sequence shown in SEQ ID NO:2 forms；
(b) by the amino acid sequence of SEQ ID NO:2 by one or several amino acid residues substitution and/or missing and/or
Addition and the protein as derived from SEQ ID NO:2 relevant to the regulation of barley moisture-proof.
2. encoding the gene of albumen described in claim 1.
3. gene according to claim 2, it is characterised in that: the gene is any in following (a1)-(a3)
DNA molecular；
(a1) DNA molecular shown in SEQ ID NO:1；
(a2) hybridize and the DNA molecular of encoding barley moisture-proof modulin with (a1) DNA sequence dna limited under strict conditions；
(a3) at least have 70% with (a1) DNA sequence dna limited, at least have 75%, at least with 80%, at least with 85%, extremely
Less with 90%, at least with 95%, at least with 96%, at least with 97%, at least with 98% or at least have it is 99% homologous
The DNA molecular of property and encoding barley moisture-proof modulin.
4. expression cassette, recombinant vector, recombinant microorganism or transgenic cell line containing gene described in Claims 2 or 3.
5.(b1) or the application of (b2) or (b3):
(b1) albumen described in claim 1, or, gene described in Claims 2 or 3, or, containing gene described in Claims 2 or 3
Expression cassette, recombinant vector, recombinant microorganism or transgenic cell line, anti-barley wet injury stress in application；
(b2) albumen described in claim 1, or, gene described in Claims 2 or 3, or, containing gene described in Claims 2 or 3
Expression cassette, recombinant vector, recombinant microorganism or transgenic cell line, cultivating barley, answering in arabidopsis moisture-proof new varieties
With；
(b3) albumen described in claim 1, or, gene described in Claims 2 or 3, or, containing gene described in Claims 2 or 3
Expression cassette, recombinant vector, recombinant microorganism or transgenic cell line, anti-arabidopsis wet injury stress in application.
6. a kind of method for cultivating plant moisture-proof kind, which is characterized in that gene described in claim 2 or 3 is transferred to purpose
Plant or expression cassette as claimed in claim 4, recombinant vector convert purpose plant, obtain genetically modified plants；The transgenosis is planted
The wet injury stress resistance of object is higher than the purpose plant.
7. the method according to claim 6 for cultivating plant moisture-proof kind, which is characterized in that the purpose plant is big
Wheat, arabidopsis.
8. a kind of method for cultivating plant moisture-proof kind according to claim 6, which is characterized in that it is characterized in that, institute
The Expressed in Transgenic Plant stated barley moisture-proof described in claim 1 regulates and controls GAP-associated protein GAP.
Priority Applications (1)
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| CN112521475A (en) * | 2020-12-25 | 2021-03-19 | 山东农业大学 | Wheat TaLAX1-A gene and application thereof in improving wheat immature embryo regeneration efficiency |
| CN114107333A (en) * | 2021-10-27 | 2022-03-01 | 上海市农业科学院 | Application of barley receptor kinase HvSERK1 in root hair growth |
| CN114874302A (en) * | 2022-05-27 | 2022-08-09 | 盐城师范学院 | Barley moisture-resistant gene HvMADS1, protein and application thereof in moisture stress resistance |
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