CN109678940A - Protein B hDnaJ6 and its encoding gene and application - Google Patents

Protein B hDnaJ6 and its encoding gene and application Download PDF

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CN109678940A
CN109678940A CN201710969733.XA CN201710969733A CN109678940A CN 109678940 A CN109678940 A CN 109678940A CN 201710969733 A CN201710969733 A CN 201710969733A CN 109678940 A CN109678940 A CN 109678940A
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bhdnaj6
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CN109678940B (en
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邓馨
刘杰
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Institute of Botany of CAS
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    • C12N15/82—Vectors or expression systems specially adapted for eukaryotic hosts for plant cells, e.g. plant artificial chromosomes (PACs)
    • C12N15/8241—Phenotypically and genetically modified plants via recombinant DNA technology
    • C12N15/8261—Phenotypically and genetically modified plants via recombinant DNA technology with agronomic (input) traits, e.g. crop yield
    • C12N15/8271—Phenotypically and genetically modified plants via recombinant DNA technology with agronomic (input) traits, e.g. crop yield for stress resistance, e.g. heavy metal resistance
    • C12N15/8273—Phenotypically and genetically modified plants via recombinant DNA technology with agronomic (input) traits, e.g. crop yield for stress resistance, e.g. heavy metal resistance for drought, cold, salt resistance
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    • C07—ORGANIC CHEMISTRY
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    • C07K2319/00—Fusion polypeptide
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    • C07K2319/20—Fusion polypeptide containing a tag with affinity for a non-protein ligand
    • C07K2319/21—Fusion polypeptide containing a tag with affinity for a non-protein ligand containing a His-tag
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    • C07K2319/00—Fusion polypeptide
    • C07K2319/20—Fusion polypeptide containing a tag with affinity for a non-protein ligand
    • C07K2319/22—Fusion polypeptide containing a tag with affinity for a non-protein ligand containing a Strep-tag
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K2319/00—Fusion polypeptide
    • C07K2319/40—Fusion polypeptide containing a tag for immunodetection, or an epitope for immunisation
    • C07K2319/41—Fusion polypeptide containing a tag for immunodetection, or an epitope for immunisation containing a Myc-tag

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Abstract

The invention discloses a kind of protein B hDnaJ6 and its encoding gene and applications.Protein provided by the invention is obtained from rotation capsule lettuce tongue fur, is named as BhDnaJ6, is following (a) or (b): (a) protein that the amino acid sequence shown in sequence 1 in sequence table forms；(b) by the amino acid sequence of sequence 1 by the substitution and/or deletion and/or addition of one or several amino acid residues and the protein as derived from sequence 1 relevant to plant drought resistance.The gene (BhDnaJ6 gene) for encoding the BhDnaJ6 albumen also belongs to protection scope of the present invention.By the channel genes wildtype Arabidopsis thaliana, what available drought resistance significantly improved turns BhDnaJ6 arabidopsis.The new varieties such as crop, the woods grass that the present invention improves cultivation drought resistance have important theory and practical significance, the cultivation and identification of resistance plant kind needed for can be used for farming and animal husbandry and ecological environment treatment.

Description

Protein B hDnaJ6 and its encoding gene and application
Technical field
The present invention relates to field of biotechnology, and in particular to a kind of protein B hDnaJ6 and its encoding gene and application.
Background technique
Arid has become the serious worldwide problem for restricting agricultural production, and the change of weather will bring it is more frequent Arid, therefore, the drought resistance that crop is improved using Protocols in Molecular Biology become one of important scientific issues urgently to be resolved.
Resurrection plant is lived in mostly in frequent drought environment, and nutritive issue can be resistant to severe drought, is met fast after water Quick-recovery normal condition.The resurrection plant having now been found that is mainly distributed in moss and pteridophyte, only sends out in angiosperm It is 135 kinds existing, it is distributed mainly on eastern Africa and south, Australia and South American region, it is scattered to be distributed in East Asia and Balkan half Island.Rotation capsule lettuce tongue fur (Boeahygrometrica) is distributed across a kind of Gesneriaceae resurrection plant of China, and popular name hygrometric boea herb turns over Soul grass, cat ear etc..Not only a part of the drought-resistant dehydration of whole plant or even excised leaf or blade is resistant to the plant Drought, and recover after meeting water, normal life state can be restored in several days.Moreover, revolving capsule lettuce tongue fur chloroplaset during the dehydration process Structure is still kept completely, and chlorophyll content is basically unchanged, and thylakoid Pigment-protein Complexes keep stable structure.
Summary of the invention
The object of the present invention is to provide a kind of protein B hDnaJ6 and its encoding gene and applications.
Protein provided by the invention is obtained from rotation capsule lettuce tongue fur, is named as BhDnaJ6, is following (a) or (b):
(a) protein that the amino acid sequence shown in sequence 1 in sequence table forms；
(b) amino acid sequence of sequence 1 by the substitution of one or several amino acid residues and/or missing and/or is added Add and the protein as derived from sequence 1 relevant to plant drought resistance.
In order to make BhDnaJ6 albumen in (a) convenient for purifying and detection, can in as sequence table amino shown in sequence 1 The amino terminal or carboxyl terminal of the protein of acid sequence composition connect upper label as shown in Table 1.
The sequence of 1 label of table
Label Residue Sequence
Poly-Arg 5-6 (usually 5) RRRRR
Poly-His 2-10 (usually 6) HHHHHH
FLAG 8 DYKDDDDK
Strep-tag II 8 WSHPQFEK
c-myc 10 EQKLISEEDL
BhDnaJ6 albumen in above-mentioned (b) can be artificial synthesized, can also first synthesize its encoding gene, then carry out biological expression It obtains.The encoding gene of BhDnaJ6 albumen in above-mentioned (b) can be by will lack in DNA sequence dna shown in sequence 2 in sequence table The codon of one or several amino acid residues, and/or carry out the missense mutation of one or several base-pairs, and/or its 5 ' The coded sequence that end and/or 3 ' ends connect label shown in table 1 obtains.
The gene (BhDnaJ6 gene) for encoding the BhDnaJ6 albumen also belongs to protection scope of the present invention.
The BhDnaJ6 gene is any DNA molecular in following (1)-(3):
(1) code area DNA molecular as shown in sequence 2 in sequence table；
(2) hybridize and encode and the DNA of plant drought resistance GAP-associated protein GAP with the DNA sequence dna that (1) limits under strict conditions Molecule；
(3) there is 90% or more homology and coding egg related with plant drought resistance to the DNA sequence dna that (1) or (2) limits White DNA molecular.
Above-mentioned stringent condition can be for 0.1 × SSPE (or 0.1 × SSC), the solution of 0.1%SDS be miscellaneous in DNA or RNA It hands over and hybridizes at 65 DEG C in experiment and wash film.
Recombinant expression carrier, expression cassette, transgenic cell line or recombinant bacterium containing the BhDnaJ6 gene belong to this The protection scope of invention.
The recombinant expression carrier of BhDnaJ6 gene can be contained with existing plant expression vector construction.The plant expression Carrier includes double base agrobacterium vector and the carrier etc. that can be used for plant micropellet bombardment.Use the gene constructed recombination table of BhDnaJ6 When up to carrier, it can be opened before its transcription initiation nucleotide plus any enhanced, composing type, organizing specific type or induction type Mover, they can be used alone or are used in combination with other plant promoters；In addition, using the gene constructed recombination of BhDnaJ6 When expression vector, also enhancer, including translational enhancer or transcriptional enhancer, these enhancer regions can be used to can be ATG Initiation codon or neighboring region initiation codon etc., but must be identical as the reading frame of coded sequence, to guarantee entire sequence Correct translation.The source of the translation control signal and initiation codon be it is extensive, can be natural, be also possible to close At.Translation initiation region can come from transcription initiation region or structural gene.For the ease of to transgenic plant cells or plant Object is identified and is screened, and can be processed to plant expression vector used, and the expression in plant, which is such as added, can produce color change The enzyme of change or gene, resistant antibiotic marker or the anti-chemical reagent marker gene of luminophor etc..From turn The security consideration of gene plant can also be added without any screening-gene, directly be screened by environment stress.
The recombinant expression carrier concretely passes through the method for the homologous recombination insetion sequence table into pLEELA carrier The recombinant plasmid for holding DNA molecular shown in 1-471 nucleotide to obtain from 5 ' of sequence 2.
The present invention also protects the application of BhDnaJ6 albumen or BhDnaJ6 gene in regulation plant drought resistance.
The plant is monocotyledon or dicotyledon.The dicotyledon can be Chinese lime mesh plant.The mountain Mandarin orange mesh plant can be crucifer.The crucifer can be Nan Jie race plant.Nan Jie race plant can be quasi- Arabis plant.The Arabidopsis plant concretely arabidopsis, such as Columbia ecotype arabidopsis.
The present invention also protects a kind of method for cultivating genetically modified plants, is obtained in BhDnaJ6 channel genes purpose plant To genetically modified plants；The genetically modified plants drought resistance is higher than the purpose plant.
In the method, the BhDnaJ6 gene can import purpose by any description above recombinant expression carrier and plant Object.The recombinant expression carrier can pass through Ti-plasmids, Ri plasmid, plant viral vector, directly delivered DNA, microinjection, electricity Lead, the conventional biology methods such as mediated by agriculture bacillus are transformed into plant cell or tissue in.
The purpose plant is monocotyledon or dicotyledon.The dicotyledon can be Chinese lime mesh plant.Institute Stating Chinese lime mesh plant can be crucifer.The crucifer can be Nan Jie race plant.Nan Jie race plant can For Arabidopsis plant.The Arabidopsis plant concretely arabidopsis, such as Columbia ecotype arabidopsis.
The present invention also protects a kind of method for improving plant drought resistance, is to improve BhDnaJ6 albumen described in purpose plant Expression quantity and/or activity, obtain drought resistance raising plant.
The present invention also protects the BhDnaJ6 albumen, the BhDnaJ6 gene or any description above method to educate in plant Application in kind.
The purpose of the breeding is the high plant of breeding drought resistance.
The plant is monocotyledon or dicotyledon.The dicotyledon can be Chinese lime mesh plant.The mountain Mandarin orange mesh plant can be crucifer.The crucifer can be Nan Jie race plant.Nan Jie race plant can be quasi- Arabis plant.The Arabidopsis plant concretely arabidopsis, such as Columbia ecotype arabidopsis.
The present invention obtains one by drought-induced expression from resurrection plant rotation capsule lettuce tongue fur (Boeahygrometrica) BhDnaJ6 gene obtains the channel genes wildtype Arabidopsis thaliana to turn BhDnaJ6 arabidopsis, compared with wildtype Arabidopsis thaliana, The drought resistance for turning BhDnaJ6 arabidopsis significantly improves, and illustrates that BhDnaJ6 is albumen relevant to plant drought.The present invention for Cultivating the new varieties such as crop, the woods grass that drought resistance improves has important theory and practical significance, can be used for farming and animal husbandry and ecology The cultivation and identification of resistance plant kind needed for environmental improvement.
Detailed description of the invention
Fig. 1 is using fluorescence quantitative PCR detection BhDnaJ6 gene in embodiment 2 in rotation capsule lettuce tongue fur (Boeahygrometrica) expression in arid resuscitation process.
Fig. 2 is subcellular localization of the BhDnaJ6 albumen in cell in embodiment 3.
Fig. 3 is T in embodiment 43In generation, turns the RT-PCR testing result of BhDnaJ6 arabidopsis.
Fig. 4 is wild type and T in embodiment 43In generation, turns the drought resisting phenotypic analysis of BhDnaJ6 arabidopsis.
Fig. 5 is wild type and T in embodiment 43In generation, turns final survival rate statistics after BhDnaJ6 arabidopsis Drying and rewatering.
Specific embodiment
Embodiment below facilitates a better understanding of the present invention, but does not limit the present invention.Experiment in following embodiments Method is unless otherwise specified conventional method.Test material as used in the following examples is unless otherwise specified certainly What routine biochemistry reagent shop was commercially available.Quantitative test in following embodiment is respectively provided with three repeated experiments, as a result makes even Mean value.
Rotation capsule lettuce tongue fur: picking up from Beijing Botanical Garden cherry ditch, by being accredited as rotation capsule lettuce tongue fur；After being selfed for 3 generations, greenhouse earth culture.
PEXSG carrier: Invitrogen company.
PLEELA carrier: Invitrogen company.
Carrier pENTR D-TOPO:Invitrogen company.
Agrobacterium tumefaciems GV3101: bibliography: Agrobacterium tumefaciens strain GV3101；Text It offers: Binary Agrobacterium vectors for plant transformation, M Bevanin, Nucleic Acids Research(1984)12(22):8711-8721；The public can obtain from Institute of Botany, Chinese Academy of Sciences.
Arabidopsis Col-0: bibliography: Arabidopsis, a useful weed.Meyerowitz EM, Cell (1989):263-270；The public can obtain from Institute of Botany, Chinese Academy of Sciences.
The acquisition of embodiment 1, protein B hDnaJ6 and its encoding gene
The total serum IgE of rotation capsule lettuce tongue fur blade is extracted, and reverse transcription is cDNA.By a large amount of sequences analysis, expression analysis with Functional verification has found a DNA encoding sequence from cDNA, as shown in the sequence 2 of sequence table, the protein such as sequence of coding Shown in the sequence 1 of list.
Protein shown in sequence 1 by sequence table is named as BhDnaJ6 albumen, is made of 156 amino acid residues.It will The unnamed gene for encoding BhDnaJ6 albumen is BhDnaJ6 gene, and open reading frame is as shown in the sequence 2 of sequence table.
The expression of embodiment 2, BhDnaJ6 gene in rotation capsule lettuce dried lactuca drought resuscitation process
1, Osmotic treatment is carried out to the rotation capsule lettuce tongue fur plant of normal growth in soil, acquires 4 kinds of samples, is respectively as follows:
(1) in soil normal growth plant (CK)；
(2) plant of normal growth stops watering Progressive drought, the plant (D5) when 5 days arid in soil；
(3) plant of normal growth stops watering Progressive drought, the plant (D14) when 14 days arid in soil；
(4) restart to water by 14 days arid plant, the plant (A) when watering 3 days.
2, after completing step 1, the rotation capsule lettuce tongue fur blade total serum IgE of each processing group is extracted, and reverse transcription is cDNA.
3, the cDNA obtained using step 2 is template, using the method detection BhDnaJ6 gene of qRT-PCR in different disposal Expression (using 18SrRNA gene as reference gene) in group rotation capsule lettuce tongue fur, using primer BhDnaJ6-F and primer The expression of the primer pair detection BhDnaJ6 gene of BhDnaJ6-R composition, the primer formed using primer 18S-F and primer 18S-R Expression to detection 18SrRNA gene.
BhDnaJ6-F:5 '-TCCGACGAATGTCTGCTTT-3 '；
BhDnaJ6-R:5 '-ACGAGGCGTTGGAATGAA-3 '；
18S-F:5 '-CTTAGTTGGTGGAGCGATTTG-3 '；
18S-R:5 '-CCTGTTATTGCCTCAAACTTCC-3 '.
As a result as shown in Figure 1.The result shows that BhDnaJ6 gene is obviously by drought-induced.
The subcellular localization of embodiment 3, BhDNAJ6 albumen in cell
1, fusion expression vector pEXSG-35S-BhDnaJ6-YFP: in the I restriction enzyme site insetion sequence of Hpa of carrier pENSG The sequence 2 of table holds DNA molecular shown in 1-468 nucleotide from 5 ', obtains fusion expression vector pEXSG-35S- BhDnaJ6-YFP (sequence verification).
2, the fusion expression vector pEXSG-35S-BhDnaJ6-YFP for obtaining step 1 converts Agrobacterium tumefaciems GV3101, Obtain recombinational agrobacterium.
3, using floral organ infusion method (bibliography: Clough SJ, Bent AF (1998) .Floral dip:a simplified method for Agrobacterium-mediated transformation of Arabidopsis Thaliana.Plant J16,735-743) Agrobacterium that obtains step 2 infects arabidopsis Col-0, obtain T0For transgenosis Arabidopsis seed.By T0The MS solid medium containing 10 μ g/mL cremarts is equably seeded in for transgenic arabidopsis seed On, resistant surviving seedling is moved into hot-house culture (22 DEG C of cultivation temperature, 16h illumination/8h is dark), obtains T1In generation, turns base Because of arabidopsis seed.By T1It is seeded in the MS culture medium solid containing 10 μ g/mL cremarts, will divide for transgenic arabidopsis seed From than the T for 3:11Hot-house culture is moved to for surviving seedling, collects T2For transgenic arabidopsis seed.By T2For the quasi- south of transgenosis Canola seed is seeded in the MS solid medium containing 10 μ g/mL cremarts, by the T of all strains of display resistance2It is moved for seedling To hot-house culture, transgenic homozygous strain arabidopsis T is obtained3For seed, T is obtained by culture3For transgenic arabidopsis.
4, the T that step 3 is obtained by two-photon laser flying-spot microscope3It is observed for transgenic arabidopsis, as a result As shown in Figure 2.The result shows that BhDnaJ6 is positioned in chloroplaset when stablizing expression in arabidopsis.
Embodiment 4, BhDnaJ6 are improving the application in plant drought ability
One, the acquisition of over-express vector is recombinated
1, the total serum IgE of rotation capsule lettuce tongue fur blade is extracted, and reverse transcription is cDNA.
2, cDNA is obtained as template using step 1, is carried out using primer BhDnaJ6-CDS-F and primer BhDnaJ6-CDS-R PCR amplification obtains amplified production.
BhDnaJ6-CDS-F:5 '-CACCTTCAGCAATGTCTTCGATAC-3 '；
BhDnaJ6-CDS-R:5 '-CTACCAGCACTGTTCAGTTTCCC-3 '.
3, it is reacted by BP, the amplified production that step 2 is obtained imports carrier pENTR D-TOPO, obtains containing ordered list Sequence 2 from 5 ' end 1-471 nucleotide shown in DNA molecular positive Entry clone plasmids pENTR-BhDnaJ6 ( Through sequence verification).
BP reaction system: 1.0 μ L, Salt solution of pcr amplification product, 0.5 μ L, pENTR D-TOPO carrier, 0.5 μ L。
BP reaction condition: 22.5 DEG C of connection reactions overnight.
4, the positive Entry clone plasmids pENTR-BhDnaJ6 that step 3 obtains is taken, LR is carried out with carrier pLEELA and reacts, Obtain the sequence 2 containing ordered list holds the 35S:BhDnaJ6 of DNA molecular shown in 1-471 nucleotide to be overexpressed from 5 ' Carrier (sequence verification).
LR reaction system: 2.0 μ L, pLEELA carrier of pENTR D-TOPO carrier, 1.0 μ L, LR mix, 0.5 μ L
LR reaction condition: in 22.5 DEG C of connection reactions overnight.
Product in above-mentioned reaction is all from the Gateway@Enzyme Mix product of Invitrogen company.
Two, the building of transgenic arabidopsis
1, the 35S:BhDnaJ6 over-express vector for obtaining step 1 imports Agrobacterium tumefaciems GV3101, obtains recombination agriculture Bacillus.
2, using floral organ infusion method (bibliography: (Clough SJ, Bent AF (1998) .Floral dip:a simplified method for Agrobacterium-mediated transformation of Arabidopsis Thaliana.Plant J 16,735-743), arabidopsis Col-0 is infected with the recombinational agrobacterium that step 1 obtains, obtains T0Generation Transgenic arabidopsis seed.
3, the T for taking step 2 to obtain0It is equably seeded in for transgenic arabidopsis seed containing 50 μ g/mL kanamycins On MS solid medium, resistant surviving seedling is moved into hot-house culture (22 DEG C of cultivation temperature, 16h illumination/8h is dark), Obtain T1For transgenic arabidopsis seed.By T1The culture of the MS containing 50 μ g/mL kanamycins is seeded in for transgenic arabidopsis seed In base solid, the T for being 3:1 by segregation ratio1Hot-house culture is moved to for surviving seedling, collects T2For transgenic arabidopsis seed.By T2 It is seeded in the MS solid medium containing 50 μ g/mL kanamycins for transgenic arabidopsis seed, the T that there will be no separation2Generation Surviving seedling moves to hot-house culture, obtains T3For transgenic arabidopsis seed, T is obtained by culture3For transgenic arabidopsis.
4, the T obtained from step 33For three strains (OE-9, OE-10 and OE-12) are selected in transgenic arabidopsis, extract Primer BhDnaJ6-RT-F and primer BhDnaJ6-RT-R composition is respectively adopted using the total serum IgE as template in blade total serum IgE Primer pair and the primer pair of primer Actin-F and primer Actin-R composition carry out RT-PCR amplification (using Actin gene as internal reference Gene)；Wildtype Arabidopsis thaliana Col-0 blade total serum IgE is also similarly expanded simultaneously；Amplified production is subjected to electrophoresis sight It examines.
BhDnaJ6-RT-F:5 '-TCCGACGAATGTCTGCTT-3 '；
BhDnaJ6-RT-R:5 '-ACGAGGCGTTGGAATGAA-3 '；
Actin-F:5 '-GTATGGTGAAGGCTGGATTTGC-3 '；
Actin-R:5 '-TG AGGTAATCAGTAAGGTCACGTCC-3 '.
As a result as shown in Figure 3.The result shows that do not amplify band in wildtype Arabidopsis thaliana (WT), OE-9, OE-10 and All there is band in OE-12, illustrates that OE-9, OE-10 and OE-12 are positive transgenic arabidopsis homozygote strain.
Three, turn the building of empty carrier arabidopsis
35S:BhDnaJ6 over-express vector is substituted using pLEELA carrier, is operated according to step 2, obtains turning zero load Body arabidopsis.
Four, drought resistance detects
Plant to be measured are as follows: wildtype Arabidopsis thaliana (WT), T3For transgenic arabidopsis (OE-9, OE-10 and OE-12) and turn sky Carrier arabidopsis.
1, plant seed to be measured is seeded on 1/2MS solid medium, 4 DEG C after Stratificated treatment 3 days, move to 22 ± 3 DEG C, It cultivates 5 days under 50% humidity, 16h illumination/8h dark condition, then arabidopsis is transferred in soil (after Nutrition Soil sterilizes It is packed into back cut diameter after mixing according to the ratio of volume ratio 1:2 with vermiculite as 7cm, in the small flower of a height of 8cm, every basin is equal 45g is filled, then flowerpot is placed in pallet, 4L water is added in pallet, sufficiently impregnates in 4h, is sealed with preservative film), 22 It is raised after being cultivated 5 days at ± 3 DEG C, continues culture 20 days, during which paid attention to guaranteeing that the supplies such as moisture are abundant, keep its growth consistent, no It is coerced.
2, the Arabidopsis thaliana Seedlings for taking step 1 to cultivate, selection is of the same size to be grouped processing, experimental group: stopping watering 21 days, phenotype was observed at arid 0 day, arid 9 days and arid 21 days and is taken a picture；Subsequent rehydration is observed phenotype and is shone after 3 days Phase, while counting survival rate；Control group: positive row culture keeps moisture sufficient, observes phenotype in same time in experimental group and shines Phase.Every group 6 plants of every plant.
As a result as shown in Figure 4 and Figure 5.Fig. 4 is Phenotypic Observation result.Fig. 5 is survival rate statistical result.
The result shows that Osmotic treatment is observed after 21 days, wildtype Arabidopsis thaliana and transgenic arabidopsis seedling are raw in control group Length is almost the same, and in experimental group, there is serious dehydration, wilting and crimp in the yellow leaf of wildtype Arabidopsis thaliana, leaf Handle loses enabling capabilities, entire lotus throne leaf can not normal extension, show approximate withered state, transgenic arabidopsis strain (OE- 9, OE-10 and OE-12) growing state be substantially better than wild type, especially OE-9 and the blade of OE-12 still keeps preferably stretching Exhibition state, the phenomenon that not wilting, crimp.It is identical as wildtype Arabidopsis thaliana to turn empty carrier arabidopsis phenotype.
After Osmotic treatment 21 days, rehydration is carried out to processing group, arabidopsis survival rate is observed and counted after 3 days, is found wild Type arabidopsis is all dead, and the survival rate of OE-9 and OE-12 is 100%, and the survival rate of OE-10 is 80%.Turn empty carrier Arabidopsis survival rate statistical result is identical as wildtype Arabidopsis thaliana.
In conclusion the drought resistance of BhDnaJ6 gene overexpression plant is substantially better than the plant of non-transgenosis, explanation BhDnaJ6 is albumen relevant to plant drought.
<110>Institute of Botany, Chinese Academy of Sciences
<120>protein B hDnaJ6 and its encoding gene and application
<160> 2
<210> 1
<211> 156
<212> PRT
<213>capsule lettuce tongue fur is revolved
<400> 1
Met Ser Ser Ile Ser Ser Phe Thr Pro Ser Glu Ile Thr Gly Arg Arg
1 5 10 15
Ile Ala Ala Val Pro Val Pro His Ile Pro Thr Asn Val Cys Phe Pro
20 25 30
His Gln Leu Arg Ile Ser Ala Gly Tyr Ser Thr Ala Asp Gln Arg Ala
35 40 45
Lys Glu Thr Ser Leu Gln Thr Gly Ser Leu Tyr Glu Ile Leu Gly Ile
50 55 60
His Ser Asn Ala Ser Cys Gln Glu Ile Lys Ser Ala Tyr Arg Lys Val
65 70 75 80
Ala Arg Leu Leu His Pro Asp Val Ala Ser Asn Ser Lys Gly Gly Gly
85 90 95
Ala Thr Thr Glu Glu Phe Met Arg Leu His Ala Ala Tyr Ala Thr Leu
100 105 110
Ser Asp Pro Glu Lys Arg Ala Met Tyr Asp Val Thr Leu Ser Arg Arg
115 120 125
Arg Arg Arg Glu Ala Arg Leu Ala Ala Ser Ser Phe Pro Glu Val Glu
130 135 140
Gly Arg Arg Arg Thr Trp Glu Thr Glu Gln Cys Trp
145 150 155
<210> 2
<211> 471
<212> DNA
<213>capsule lettuce tongue fur is revolved
<400> 2
atgtcttcga tatcatcgtt cacaccgagt gaaattacag gccgccgaat cgccgccgtg 60
cctgttccac atattccgac gaatgtctgc tttccacacc agctccgaat ctccgccggt 120
tactccacag ctgatcagag ggctaaagaa acatctctgc agaccggatc attgtacgaa 180
atcctgggga ttcattccaa cgcctcgtgt caagagatca agtcggcgta tagaaaagtg 240
gccagactgc tgcatccgga cgtcgcatcc aattccaaag gcggaggagc tactactgaa 300
gagtttatga gactgcacgc ggcgtatgct actctctccg accctgagaa gcgcgcgatg 360
tatgatgtga cgctttccag acgacggcgg agggaggcgc gcttggcggc gagttctttt 420
ccggaggtgg aagggaggag gcggacttgg gaaactgaac agtgctggta g 471

Claims (10)

  1. It is following (a) or (b) 1. a kind of protein:
    (a) protein that the amino acid sequence shown in sequence 1 in sequence table forms；
    (b) by the amino acid sequence of sequence 1 by one or several amino acid residues substitution and/or deletion and/or addition and The protein as derived from sequence 1 relevant to plant drought resistance.
  2. 2. encoding the gene of protein described in claim 1.
  3. 3. gene as claimed in claim 2, it is characterised in that: the gene is any the DNA points in following (1)-(3) Son:
    (1) code area DNA molecular as shown in sequence 2 in sequence table；
    (2) hybridize and encode and the DNA molecular of plant drought resistance GAP-associated protein GAP with the DNA sequence dna that (1) limits under strict conditions；
    (3) DNA sequence dna limited with (1) or (2) has 90% or more homology and encodes and plant drought resistance GAP-associated protein GAP DNA molecular.
  4. 4. recombinant expression carrier, expression cassette, transgenic cell line or recombinant bacterium containing gene described in Claims 2 or 3.
  5. 5. protein described in claim 1, or, gene described in Claims 2 or 3, the application in regulation plant drought resistance.
  6. 6. a kind of method for cultivating genetically modified plants, is turned in channel genes purpose plant described in Claims 2 or 3 Gene plant；The genetically modified plants drought resistance is higher than the purpose plant.
  7. 7. a kind of method for improving plant drought resistance, be improve in purpose plant the expression quantity of protein described in claim 1 and/ Or activity, obtain the plant of drought resistance raising.
  8. 8. method according to claim 6 or 7, it is characterised in that: the purpose plant is arabidopsis.
  9. 9. protein described in claim 1, or, gene described in Claims 2 or 3, or, any side of claim 6 to 8 Method, the application in plant breeding.
  10. 10. application as claimed in claim 9, it is characterised in that: the purpose of the breeding is the high plant of breeding drought resistance.
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