CN109678940B - Protein BhDnaJ6, and coding gene and application thereof - Google Patents

Abstract

The invention discloses a protein BhDnaJ6, and a coding gene and application thereof. The protein provided by the invention, named as BhDnaJ6, is (a) or (b) as follows: (a) a protein consisting of an amino acid sequence shown in a sequence 1 in a sequence table; (b) and (b) a protein which is obtained by substituting and/or deleting and/or adding one or more amino acid residues in the amino acid sequence shown in the sequence 1, is related to the drought resistance of plants and is derived from the sequence 1. The gene for coding the BhDnaJ6 protein (BhDnaJ6 gene) also belongs to the protection scope of the invention. The gene is introduced into wild arabidopsis thaliana to obtain transgenic BhDnaJ6 arabidopsis thaliana with obviously improved drought resistance. The invention has important theoretical and practical significance for cultivating new varieties of crops, forest and grass and the like with improved drought resistance, and can be used for cultivating and identifying resistant plant varieties required by agriculture and animal husbandry and ecological environment management.

Description

Protein BhDnaJ6 and its encoding gene and application

technical field

The invention relates to the field of biotechnology, in particular to a protein BhDnaJ6 and its encoding gene and application.

Background technique

Drought has become a worldwide problem that severely restricts agricultural production, and climate change will bring more frequent droughts. Therefore, the use of molecular biology techniques to improve the drought resistance of crops has become one of the important scientific issues to be solved urgently.

Most of the resuscitated plants live in frequent drought environments, and their vegetative tissues can tolerate severe drought and quickly return to normal after encountering water. The recovered plants that have been found are mainly distributed in mosses and ferns, and only 135 species have been found in angiosperms, mainly distributed in eastern and southern Africa, Australia and South America, and scattered in East Asia and the Balkans. Boeahygrometrica (Boeahygrometrica) is a resuscitated plant of the Gesneriaceae family distributed in China. The plant is not only resistant to drought and dehydration of the whole plant, but even the isolated leaves or a part of the leaves can be drought-tolerant, and can be recovered after encountering water, and can return to a normal living state within a few days. In addition, the chloroplast structure of Capsula citrifolia remained intact during the dehydration process, the chlorophyll content was basically unchanged, and the thylakoid pigment protein complex remained stable in structure.

SUMMARY OF THE INVENTION

The purpose of the present invention is to provide a protein BhDnaJ6 and its encoding gene and application.

The protein provided by the present invention, obtained from the spinosa, named BhDnaJ6, is as follows (a) or (b):

(a) a protein consisting of the amino acid sequence shown in Sequence 1 in the Sequence Listing;

(b) A protein derived from SEQ ID NO: 1 wherein the amino acid sequence of SEQ ID NO: 1 has undergone substitution and/or deletion and/or addition of one or several amino acid residues and is related to plant drought resistance.

In order to facilitate purification and detection of the BhDnaJ6 protein in (a), a tag as shown in Table 1 can be attached to the amino terminus or carboxyl terminus of the protein consisting of the amino acid sequence shown in SEQ ID NO: 1 in the sequence listing.

Table 1 Sequences of tags

Label Residues 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

The BhDnaJ6 protein in (b) above can be obtained by artificial synthesis, or by first synthesizing its encoding gene and then biologically expressing it. The coding gene of the BhDnaJ6 protein in the above (b) can be obtained by deleting the codons of one or several amino acid residues in the DNA sequence shown in SEQ ID NO: 2 in the Sequence Listing, and/or making one or several base pair errors. A sense mutation, and/or the coding sequence of the tag shown in Table 1 is attached to its 5' end and/or 3' end.

The gene encoding the BhDnaJ6 protein (BhDnaJ6 gene) also belongs to the protection scope of the present invention.

The BhDnaJ6 gene is the DNA molecule described in any one of the following (1)-(3):

(1) a DNA molecule whose coding region is shown in sequence 2 in the sequence listing;

(2) a DNA molecule that hybridizes with the DNA sequence defined in (1) under stringent conditions and encodes a protein related to plant drought resistance;

(3) A DNA molecule that has more than 90% homology with the DNA sequence defined in (1) or (2) and encodes a protein related to plant drought resistance.

The above stringent conditions can be used in DNA or RNA hybridization experiments using a solution of 0.1×SSPE (or 0.1×SSC), 0.1% SDS, hybridization and membrane washing at 65° C. in DNA or RNA hybridization experiments.

Recombinant expression vectors, expression cassettes, transgenic cell lines or recombinant bacteria containing the BhDnaJ6 gene all belong to the protection scope of the present invention.

The recombinant expression vector containing the BhDnaJ6 gene can be constructed by using the existing plant expression vector. The plant expression vectors include binary Agrobacterium vectors and vectors that can be used for plant microprojectile bombardment, and the like. When using the BhDnaJ6 gene to construct a recombinant expression vector, any enhanced, constitutive, tissue-specific or inducible promoter can be added before its transcription initiation nucleotide, which can be used alone or with other plant promoters. Combined use; in addition, when using the BhDnaJ6 gene to construct a recombinant expression vector, enhancers can also be used, including translation enhancers or transcription enhancers. These enhancer regions can be ATG initiation codons or adjacent region initiation codons, etc., but Must be in the same reading frame as the coding sequence to ensure correct translation of the entire sequence. The translation control signals and initiation codons can be derived from a wide variety of sources, either natural or synthetic. The translation initiation region can be derived from a transcription initiation region or a structural gene. In order to facilitate the identification and screening of transgenic plant cells or plants, the plant expression vector used can be processed, such as adding genes that express enzymes or light-emitting compounds that can produce color changes in plants, antibiotic markers with resistance or resistance to Chemical reagents to label genes, etc. Considering the safety of transgenic plants, it is also possible to directly screen by adversity stress without adding any screening genes.

Specifically, the recombinant expression vector can be a recombinant plasmid obtained by inserting the DNA molecule represented by nucleotides 1-471 at the 5' end of sequence 2 of the sequence listing into the pLEELA vector by means of homologous recombination.

The invention also protects the application of BhDnaJ6 protein or BhDnaJ6 gene in regulating plant drought resistance.

The plants are monocotyledonous or dicotyledonous. The dicotyledonous plant may be a Caperaceae plant. The Caperaceae plant may be a cruciferous plant. The cruciferous plant may be a plant of the Arabidopsis family. The Arabidopsis plant may be an Arabidopsis plant. The Arabidopsis plant can specifically be Arabidopsis thaliana, such as Colombia ecotype Arabidopsis thaliana.

The present invention also protects a method for cultivating a transgenic plant, which is to introduce the BhDnaJ6 gene into a target plant to obtain a transgenic plant; the transgenic plant has higher drought resistance than the target plant.

In the method, the BhDnaJ6 gene can be introduced into the target plant through any of the above recombinant expression vectors. The recombinant expression vector can be transformed into plant cells or tissues by conventional biological methods such as Ti plasmid, Ri plasmid, plant virus vector, direct DNA transformation, microinjection, electrical conductivity, and Agrobacterium-mediated transformation.

The target plant is a monocotyledonous plant or a dicotyledonous plant. The dicotyledonous plant may be a Caperaceae plant. The Caperaceae plant may be a cruciferous plant. The cruciferous plant may be a plant of the Arabidopsis family. The Arabidopsis plant may be an Arabidopsis plant. The Arabidopsis plant can specifically be Arabidopsis thaliana, such as Colombia ecotype Arabidopsis thaliana.

The present invention also protects a method for improving the drought resistance of plants, which is to increase the expression and/or activity of the BhDnaJ6 protein in the target plant to obtain plants with improved drought resistance.

The present invention also protects the application of the BhDnaJ6 protein, the BhDnaJ6 gene or any of the above methods in plant breeding.

The purpose of the breeding is to select plants with high drought resistance.

The plants are monocotyledonous or dicotyledonous. The dicotyledonous plant may be a Caperaceae plant. The Caperaceae plant may be a cruciferous plant. The cruciferous plant may be a plant of the Arabidopsis family. The Arabidopsis plant may be an Arabidopsis plant. The Arabidopsis plant can specifically be Arabidopsis thaliana, such as Colombia ecotype Arabidopsis thaliana.

The invention obtains a BhDnaJ6 gene whose expression is induced by drought from the resuscitated plant Boeahygrometrica, and introduces the gene into wild-type Arabidopsis thaliana to obtain transgenic BhDnaJ6 Arabidopsis. Compared with wild-type Arabidopsis thaliana, the transgenic The drought resistance of BhDnaJ6 in Arabidopsis was significantly improved, indicating that BhDnaJ6 is a protein related to plant drought resistance. The invention has important theoretical and practical significance for cultivating new varieties of crops, forest and grass with improved drought resistance, and can be used for breeding and identification of resistant plant varieties required for agriculture and animal husbandry and ecological environment management.

Description of drawings

Figure 1 shows the expression of BhDnaJ6 gene in the process of drought recovery of Boeahygrometrica detected by fluorescence quantitative PCR in Example 2.

FIG. 2 shows the subcellular localization of BhDnaJ6 protein in cells in Example 3. FIG.

FIG. 3 is the RT-PCR detection result of Arabidopsis thaliana transduced with T ₃ generation in Example 4. FIG.

FIG. 4 is the drought resistance phenotype analysis of wild-type and T ₃ generation transgenic BhDnaJ6 Arabidopsis in Example 4. FIG.

Figure 5 shows the final survival rate statistics of wild-type and T ₃ generation transgenic BhDnaJ6 Arabidopsis in Example 4 after drought and rehydration.

Detailed ways

The following examples facilitate a better understanding of the present invention, but do not limit the present invention. The experimental methods in the following examples are conventional methods unless otherwise specified. The test materials used in the following examples were purchased from conventional biochemical reagent stores unless otherwise specified. The quantitative tests in the following examples are all set to repeat the experiments three times, and the results are averaged.

Capsule: collected from the cherry ditch of Beijing Botanical Garden, and identified as Capsule; after 3 generations of self-crossing, it was cultivated in greenhouse soil.

pEXSG vector: Invitrogen.

pLEELA vector: Invitrogen.

Vector pENTR D-TOPO: Invitrogen.

Agrobacterium tumefaciens GV3101: Reference: Agrobacterium tumefaciens strain GV3101; Literature: Binary Agrobacterium vectors for plant transformation, M Bevanin, Nucleic Acids Research (1984) 12(22): 8711-8721; publicly available from the Institute of Botany, Chinese Academy of Sciences.

Arabidopsis Col-0: Reference: Arabidopsis, a useful weed. Meyerowitz EM, Cell (1989): 263-270; publicly available from the Institute of Botany, Chinese Academy of Sciences.

Example 1. Acquisition of protein BhDnaJ6 and its encoding gene

Total RNA was extracted from the leaves of C. radix and reverse transcribed into cDNA. After extensive sequence analysis, expression analysis and functional verification, a DNA coding sequence was found from the cDNA, as shown in Sequence 2 of the Sequence Listing, and the encoded protein was shown in Sequence 1 of the Sequence Listing.

The protein shown in SEQ ID NO: 1 of the Sequence Listing is named BhDnaJ6 protein, which consists of 156 amino acid residues. The gene encoding the BhDnaJ6 protein is named BhDnaJ6 gene, and its open reading frame is shown in SEQ ID NO: 2 of the sequence listing.

Example 2. Expression of BhDnaJ6 gene in the process of recovery from drought in C.

1. Drought treatment is carried out to the normal growth of C. serrata plants in the soil, and 4 kinds of samples are collected, which are respectively:

(1) Plants growing normally in soil (CK);

(2) The plants that normally grow in the soil stop watering and gradually become dry, and the plants (D5) when they are dry for 5 days;

(3) The plants that normally grow in the soil stop watering and gradually become dry, and the plants are dry for 14 days (D14);

(4) Plants after 14 days of drought restarted watering, and plants after 3 days of watering (A).

2. After the completion of step 1, the total RNA of the leaves of C. radix in each treatment group was extracted, and reverse transcribed into cDNA.

3. Using the cDNA obtained in step 2 as the template, the expression of BhDnaJ6 gene in different treatment groups was detected by qRT-PCR (with 18SrRNA gene as the internal reference gene), using primer BhDnaJ6-F and primer BhDnaJ6- The primer pair composed of R was used to detect the expression of BhDnaJ6 gene, and the primer pair composed of primer 18S-F and primer 18S-R was used to detect the expression of 18SrRNA gene.

BhDnaJ6-F: 5'-TCCGACGAATGTCTGCTTT-3';

BhDnaJ6-R: 5'-ACGAGGCGTTGGAATGAA-3';

18S-F: 5'-CTTAGTTGGTGGAGCGATTTG-3';

18S-R: 5'-CCTGTTATTGCCTCAAACTTCC-3'.

The results are shown in Figure 1. The results showed that the BhDnaJ6 gene was obviously induced by drought.

Example 3. Subcellular localization of BhDNAJ6 protein in cells

1. Fusion expression vector pEXSG-35S-BhDnaJ6-YFP: insert the DNA molecule shown in nucleotides 1-468 from the 5' end of sequence 2 in the HpaI restriction site of the vector pENSG to obtain fusion expression Vector pEXSG-35S-BhDnaJ6-YFP (verified by sequencing).

2. Transform the fusion expression vector pEXSG-35S-BhDnaJ6-YFP obtained in step 1 into Agrobacterium tumefaciens GV3101 to obtain recombinant Agrobacterium.

3. The Agrobacterium infection obtained in step 2 was simulated by the method of soaking the flower organs (reference: Clough SJ, Bent AF (1998). Floral dip: asmplified method for Agrobacterium-mediated transformation of Arabidopsisthaliana. Plant J16, 735-743). Arabidopsis Col-0 to obtain T ₀ generation transgenic Arabidopsis seeds. The T ₀ generation transgenic Arabidopsis seeds were evenly sown on MS solid medium containing 10 μg/mL glufosinate, and the resistant surviving seedlings were moved to the greenhouse for cultivation (culturing temperature 22°C, 16h light/8h dark). , to obtain T ₁ generation transgenic Arabidopsis seeds. The T ₁ generation transgenic Arabidopsis seeds were sown in MS medium solid containing 10 μg/mL glufosinate, and the T ₁ generation surviving seedlings with a separation ratio of 3:1 were moved to the greenhouse for cultivation, and the T ₂ generation transgenic Arabidopsis was collected. mustard seeds. The T ₂ generation transgenic Arabidopsis seeds were sown in MS solid medium containing 10 μg/mL glufosinate, and the T ₂ generation seedlings of all resistant lines were transferred to the greenhouse for cultivation to obtain transgenic homozygous lines. Arabidopsis T ₃ generation seeds were cultured to obtain T ₃ generation transgenic Arabidopsis thaliana.

4. The T ₃ generation transgenic Arabidopsis obtained in step 3 was observed by a two-photon laser scanning microscope, and the results are shown in FIG. 2 . The results showed that BhDnaJ6 localized in the chloroplast when stably expressed in Arabidopsis.

Example 4. Application of BhDnaJ6 in improving plant drought resistance

First, the acquisition of recombinant overexpression vector

1. Extract the total RNA from the leaves of C. spp. and reverse-transcribe it into cDNA.

2. Using the cDNA obtained in step 1 as a template, use primer BhDnaJ6-CDS-F and primer BhDnaJ6-CDS-R to perform PCR amplification to obtain an amplification product.

BhDnaJ6-CDS-F: 5′-CACCTTCAGCAATGTCTTCGATAC-3′;

BhDnaJ6-CDS-R: 5'-CTACCAGCACTGTTCAGTTTCCC-3'.

3. The amplified product obtained in step 2 is introduced into the vector pENTR D-TOPO by BP reaction, and a positive entry clone containing the DNA molecule shown in the 1-471 nucleotides from the 5' end of SEQ ID NO: 2 of the sequence listing is obtained. Plasmid pENTR-BhDnaJ6 (verified by sequencing).

BP reaction system: PCR amplification product 1.0 μL, Salt solution 0.5 μL, pENTR D-TOPO vector 0.5 μL.

BP reaction conditions: ligation reaction at 22.5°C overnight.

4. Take the positive entry clone plasmid pENTR-BhDnaJ6 obtained in step 3, and carry out LR reaction with the vector pLEELA to obtain 35S of the DNA molecule shown in the 1-471 nucleotides from the 5' end of Sequence 2 of the sequence listing: BhDnaJ6 overexpression vector (validated by sequencing).

LR reaction system: pENTR D-TOPO vector 2.0 μL, pLEELA vector 1.0 μL, LR mix 0.5 μL

LR reaction conditions: ligation reaction at 22.5°C overnight.

The products in the above reactions are all from the Gateway@Enzyme Mix product of Invitrogen.

2. Construction of transgenic Arabidopsis

1. The 35S:BhDnaJ6 overexpression vector obtained in step 1 was introduced into Agrobacterium tumefaciens GV3101 to obtain recombinant Agrobacterium.

2. Using the soaking method of flower organs (Reference: (Clough SJ, Bent AF (1998). Floral dip: asmplified method for Agrobacterium-mediated transformation of Arabidopsisthaliana. Plant J 16, 735-743), using the recombinant Agrobacterium obtained in step 1 to infiltrate Arabidopsis Col-0 was stained to obtain T ₀ generation transgenic Arabidopsis seeds.

3. Take the T ₀ generation transgenic Arabidopsis seeds obtained in step 2 and evenly sown them on MS solid medium containing 50 μg/mL kanamycin, and move the resistant surviving seedlings to the greenhouse for cultivation (cultivation temperature 22° C.). , 16h light/8h dark) to obtain T ₁ generation transgenic Arabidopsis seeds. The seeds of T ₁ generation transgenic Arabidopsis thaliana were sown in MS medium solid containing 50 μg/mL kanamycin, and the T ₁ generation surviving seedlings with a separation ratio of 3:1 were moved to the greenhouse for culture, and the T ₂ generation transgenic seedlings were collected. Arabidopsis seeds. The T ₂ generation transgenic Arabidopsis seeds were sown in MS solid medium containing 50 μg/mL kanamycin, and the T ₂ generation surviving seedlings that did not separate were moved to the greenhouse for cultivation to obtain the T ₃ generation transgenic Arabidopsis seeds. , the T ₃ generation transgenic Arabidopsis was obtained after culture.

4. Select three lines (OE-9, OE-10 and OE-12) from the T ₃ generation transgenic Arabidopsis obtained in step 3, extract total RNA from leaves, use the total RNA as a template, and use primers respectively A primer pair consisting of BhDnaJ6-RT-F and primer BhDnaJ6-RT-R and a primer pair consisting of primer Actin-F and primer Actin-R were used for RT-PCR amplification (with Actin gene as internal reference gene); The total RNA of Arabidopsis Col-0 leaves was also amplified in the same way; the amplified products were observed by electrophoresis.

BhDnaJ6-RT-F: 5'-TCCGACGAATGTCTGCTT-3';

BhDnaJ6-RT-R: 5'-ACGAGGCGTTGGAATGAA-3';

Actin-F: 5'-GTATGGTGAAGGCTGGATTTGC-3';

Actin-R: 5'-TG AGGTAATCAGTAAGGTCACGTCC-3'.

The results are shown in Figure 3. The results showed that no bands were amplified in wild-type Arabidopsis thaliana (WT), but bands were found in OE-9, OE-10 and OE-12, indicating that OE-9, OE-10 and OE-12 were all Positive transgenic Arabidopsis homozygous line.

3. Construction of the empty vector Arabidopsis thaliana

The pLEELA vector was used to replace the 35S:BhDnaJ6 overexpression vector, and the operation was performed according to step 2 to obtain the empty vector Arabidopsis thaliana.

4. Drought resistance test

The plants to be tested are: wild-type Arabidopsis (WT), T ₃ generation transgenic Arabidopsis (OE-9, OE-10 and OE-12) and Arabidopsis thaliana with an empty vector.

1. The seeds of the plants to be tested were sown on 1/2MS solid medium, and after stratified treatment at 4°C for 3 days, moved to 22±3°C, 50% humidity, and 16h light/8h dark conditions for 5 days. Arabidopsis is transferred to the soil (the nutrient soil is sterilized and mixed with vermiculite in a volume ratio of 1:2, and then loaded into a small flower pot with an upper mouth diameter of 7cm and a height of 8cm, each pot is filled with 45g, Then put the flowerpot in the tray, add 4L water to the tray, soak it fully for 4h, seal it with plastic wrap), incubate it at 22±3℃ for 5 days, open it, and continue to cultivate for 20 days, during which time pay attention to ensure moisture, etc. Sufficient supply allows it to grow consistently without stress.

2. Take the Arabidopsis thaliana seedlings cultivated in step 1, select the same size for grouping treatment, the experimental group: stop watering for 21 days, observe the phenotype and take pictures at the time of drought 0 days, drought 9 days and drought 21 days; then rehydrate , 3 days later, the phenotype was observed and photographed, and the survival rate was counted at the same time; control group: normal culture, keeping sufficient water, the phenotype was observed and photographed at the same time in the experimental group. There are 6 plants in each group.

The results are shown in Figures 4 and 5. Figure 4 shows the phenotypic observations. Figure 5 shows the statistical results of the survival rate.

The results showed that after 21 days of drought treatment, the growth of wild-type Arabidopsis and transgenic Arabidopsis in the control group was basically the same, while in the experimental group, the leaves of the wild-type Arabidopsis turned yellow, with severe water loss, wilting and The curling phenomenon, the petioles lost their supporting ability, the whole rosette leaves could not stretch normally, showing a nearly dead state. The growth of the transgenic Arabidopsis lines (OE-9, OE-10 and OE-12) was significantly better than that of the wild type, especially The leaves of OE-9 and OE-12 still maintained a good stretched state without wilting and curling. The phenotype of the empty vector Arabidopsis is the same as that of wild-type Arabidopsis.

After 21 days of drought treatment, the treatment group was rehydrated. After 3 days, the survival rate of Arabidopsis thaliana was observed and counted. It was found that all wild-type Arabidopsis died, while the survival rates of OE-9 and OE-12 were both 100%. 10 has an 80% survival rate. The statistical results of the survival rate of Arabidopsis thaliana with the empty vector were the same as those of wild-type Arabidopsis thaliana.

In conclusion, the drought resistance of BhDnaJ6 gene overexpressing plants was significantly better than that of non-transgenic plants, indicating that BhDnaJ6 is a protein related to plant drought resistance.

<110> Institute of Botany, Chinese Academy of Sciences

<120> Protein BhDnaJ6 and its encoding gene and application

<160> 2

<210> 1

<211> 156

<212> PRT

<213> Capsule vulgaris

<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> Capsules

<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 (9)

1. the application of a protein or its encoding gene in regulating plant drought resistance; The amino acid sequence of the protein is shown in SEQ ID NO: 1 in the sequence listing. 2 . The application according to claim 1 , wherein the gene is the DNA molecule shown in sequence 2 in the sequence listing. 3 . 3. a method for cultivating a transgenic plant, comprising introducing a protein-encoding gene into a target plant to obtain a transgenic plant; the transgenic plant has a higher drought resistance than the target plant; The amino acid sequence of the protein is shown in SEQ ID NO: 1 in the sequence listing. 4 . The method according to claim 3 , wherein the gene is the DNA molecule shown in Sequence 2 in the sequence listing. 5 . 5. A method for improving plant drought resistance, which is to increase the expression and/or activity of a protein in a target plant to obtain a plant with improved drought resistance; the amino acid sequence of the protein is shown in SEQ ID NO: 1 in the sequence listing. 6. The method of any one of claims 3-5, wherein the target plant is Arabidopsis thaliana. 7. Application of a protein or its encoding gene in plant breeding; the amino acid sequence of the protein is shown in Sequence 1 in the sequence listing; the purpose of the breeding is to select plants with high drought resistance. 8 . The application according to claim 7 , wherein the gene is the DNA molecule shown in Sequence 2 in the sequence listing. 9 . 9. The application of the method of any one of claims 3-6 in plant breeding; the purpose of the breeding is to select and breed plants with high drought resistance.

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