Drought-resistant gene OsDEL1, amplification method and application of drought-resistant gene OsDEL1 in plant drought-resistant improvement

Abstract

The invention discloses a drought-resistant gene OsDEL1, an amplification method and application of the drought-resistant gene OsDEL1 in plant drought-resistant improvement, and belongs to the technical field of plant genetic engineering. The invention provides a drought-resistant gene OsDEL1. The nucleotide sequence of the drought-resistant gene OsDEL1 is as shown in SEQ ID NO. 1. The invention also provides a primer pair for amplifying the drought-resistant gene OsDEL1, a method for amplifying the drought-resistant gene OsDEL1 by using the primer pair, and an application of the drought-resistant gene OsDEL1 in improving the drought resistance of plants. The drought-resistant gene OsDEL1 and the corresponding protein are applied to genetic engineering genetic breeding of crops, a new variety of drought-resistant rice is cultivated, the safety hazard of abnormal drought caused by global climate change to food crops is reduced, and the drought-resistant gene OsDEL1 and the corresponding protein have important reference significance for related research on drought resistance of other crops.

Description

Technical Field

[0001] The invention relates to the technical field of plant genetic engineering, and in particular to a drought-resistant gene OsDEL1, an amplification method and an application thereof in improving plant drought resistance. Background Art

[0002] Rice (Oryza sativa L.) is one of the most important food crops in the world. Its production is affected by a variety of environmental factors, among which drought is one of the main abiotic stresses that limit rice yield. Therefore, improving rice drought resistance is crucial to ensuring food security. When plants perceive drought stress, they will start the expression of a series of drought stress-related genes to regulate the tolerance of plants to drought. Transcription factors play a very important role in the molecular regulatory network of plant drought resistance. They can activate or inhibit the transcription of downstream target genes in response to drought stress by binding to specific cis-acting elements of downstream target gene promoters. Among them, transcription factor families such as bZIP, NAC MYB, bHLH, WRKY, AP2 / EREBP and GRF can all participate in regulating the response of rice to adverse stress. Members of the E2F / DP transcription factor family can also participate in regulating plant drought resistance. For example, overexpression of the maize ZmE2F gene in Arabidopsis can improve the drought resistance of Arabidopsis. However, there are relatively few studies on members of this family.

[0003] Mining new rice drought-resistant genetic resources and clarifying its molecular mechanism of drought resistance can provide a theoretical basis for breeding new high-quality drought-resistant rice varieties. It is of great significance to ensure the safe production and sustainable development of rice. Since there is no relevant report on whether the OsDEL1 gene can improve the drought resistance of rice, cloning the OsDEL1 gene from rice and exploring the biological function of OsDEL1 in improving rice drought resistance are of great significance for breeding new drought-resistant rice varieties. Summary of the invention

[0004] The purpose of the present invention is to provide a drought-resistant gene OsDEL1, an amplification method and an application thereof in improving plant drought resistance, so as to solve the problems existing in the above-mentioned prior art. The OsDEL1 gene provided by the present invention can improve the resistance of rice to drought stress and has a good application prospect in breeding drought-resistant rice varieties. The present invention lays an important foundation for rice drought-resistant breeding.

[0005] To achieve the above object, the present invention provides the following solutions:

[0006] Technical solution 1: A drought-resistant gene OsDEL1, the nucleotide sequence of the drought-resistant gene OsDEL1 is shown in SEQ ID NO.1.

[0007] Technical solution 2: The protein encoded by the drought-resistant gene OsDEL1.

[0008] Furthermore, the amino acid sequence of the protein is shown in SEQ ID NO.2.

[0009] Technical solution three: a set of primer pairs for amplifying the drought resistance gene OsDEL1, including an upstream primer with a nucleotide sequence such as SEQ ID NO.3 and a downstream primer with a nucleotide sequence such as SEQ ID NO.4.

[0010] Technical solution 4: A method for amplifying the drought-resistant gene OsDEL1 using the primer pair, comprising the following steps: using a plant nucleic acid molecule as a template, preparing the template and the primer pair into a PCR amplification system for PCR amplification to obtain the drought-resistant gene OsDEL1.

[0011] Technical Solution 5: A method for improving plant drought resistance, which improves plant drought resistance by reducing the expression level of the OsDEL1 gene.

[0012] Technical solution six: Application of the drought-resistant gene OsDEL1, the protein or the drought-resistant gene OsDEL1 amplified by the method in improving plant drought resistance.

[0013] Furthermore, the plant includes rice.

[0014] Technical Solution 7: Application of the drought-resistant gene OsDEL1, the protein or the drought-resistant gene OsDEL1 amplified by the method in cultivating different drought-resistant crop germplasms.

[0015] Furthermore, the drought resistance is reduced by overexpressing the expression level of the drought resistance gene OsDEL1; and the drought resistance is enhanced by reducing the expression level of the drought resistance gene OsDEL1.

[0016] The present invention finds that knocking out the OsDEL1 gene can improve the drought resistance of rice.

[0017] The present invention provides an application of an OsDEL1 gene in regulating rice drought resistance. Specifically, overexpression of the OsDEL1 gene can reduce the survival rate of rice under drought conditions and reduce the drought resistance of rice; knocking out the OsDEL1 gene can increase the survival rate of rice under drought and improve the drought resistance of rice, and knocking out the OsDEL1 gene can increase the sensitivity of rice to ABA; the nucleotide sequence of the OsDEL1 gene is shown in SEQ ID NO.1, and the amino acid sequence of the protein encoded by the rice OsDEL1 gene is shown in SEQ ID NO.2.

[0018] The method for reducing the expression level of the OsDEL1 gene in rice is to knock out the OsDEL1 gene in rice and transfer it into wild-type rice recipient materials to improve the drought resistance of rice.

[0019] Furthermore, a knockout vector of the rice OsDEL1 gene is constructed using the CRISPR / Cas9 method, and the knockout vector is transferred into a wild-type rice recipient material to improve the drought resistance of rice.

[0020] Furthermore, the variety of rice is Zhonghua 11.

[0021] The present invention discloses the following technical effects:

[0022] The invention constructs an overexpression and gene knockout vector of rice OsDEL1 gene, and transfers the overexpression and knockout vector into rice receptor material by Agrobacterium-mediated method. Fluorescence quantitative PCR and sequencing analysis are used to detect positive plants. Drought resistance analysis is performed on the obtained positive transgenic plants, and the results show that OsDEL1 gene can improve the tolerance of rice to drought stress, knocking out the gene in rice can significantly improve the drought resistance of rice, and overexpression of OsDEL1 gene significantly reduces the drought resistance of rice. Therefore, rice OsDEL1 gene can be applied to genetic engineering breeding of crops, has good application prospects in cultivating drought-resistant rice varieties, and lays an important foundation for drought-resistant rice breeding. BRIEF DESCRIPTION OF THE DRAWINGS

[0023] In order to more clearly illustrate the embodiments of the present invention or the technical solutions in the prior art, the drawings required for use in the embodiments will be briefly introduced below. Obviously, the drawings described below are only some embodiments of the present invention. For ordinary technicians in this field, other drawings can be obtained based on these drawings without paying creative work.

[0024] Figure 1 The results of real-time fluorescence quantitative PCR analysis of the expression pattern of OsDEL1 in wild-type ZH11 at different time points after treatment with 20% PEG (A) and 50μM ABA (B); the untreated time was taken as the control and recorded as 0h; * represents a significant difference compared with the control (*, P<0.05, **, P<0.01, ***, P<0.001), and the statistical method was T-test;

[0025] Figure 2 The expression level of OsDEL1 in OsDEL1 overexpressing transgenic rice was detected by real-time fluorescence quantitative PCR; * indicates significant difference compared with the wild type (*, P<0.05, **, P<0.01, ***, P<0.001), and the statistical method was T-test;

[0026] Figure 3 The target sequence information of the first and second exon regions of the OsDEL1 gene (A) and the editing effect of the target sequence of knockout transgenic rice (B);

[0027] Figure 4 The rehydration conditions of rice in each treatment group, where A is before treatment, B is 24 hours after rehydration, and C is 8 days after rehydration;

[0028] Figure 5 The rehydration conditions of rice in each treatment group, where A is before treatment, B is 24 hours after rehydration, and C is 8 days after rehydration;

[0029] Figure 6 The survival status of rice in each treatment group under drought stress, where A is wild-type and OsDEL1-ox rice, and B is wild-type and del1 transgenic rice;

[0030] Figure 7 The water loss rate of OsDEL1 gene overexpression and knockout transgenic rice under drought stress was measured; * indicates significant difference compared with the wild type (*, P<0.05, **, P<0.01, ***, P<0.001), and the statistical method was T-test;

[0031] Figure 8 The sensitivity of wild-type rice (ZH11) and OsDEL1 gene knockout transgenic rice (del1-1 and del1-2) to ABA, where A is the germination of the CK group (0 μM ABA), and B is the germination under 5 μM ABA conditions;

[0032] Fig. 9 Figure 2 shows the statistics of ABA sensitivity in wild-type rice (ZH11) and OsDEL1 gene knockout transgenic rice (del1-1 and del1-2), where A is the root length and shoot length statistics of the CK group (0 μM ABA), and B is the root length and shoot length statistics under 5 μM ABA conditions. DETAILED DESCRIPTION

[0033] Various exemplary embodiments of the present invention will now be described in detail. This detailed description should not be considered as limiting the present invention, but should be understood as a more detailed description of certain aspects, features, and embodiments of the present invention.

[0034] It should be understood that the terms described in the present invention are only for describing a particular embodiment and are not intended to limit the present invention. In addition, for the numerical range in the present invention, it should be understood that each intermediate value between the upper and lower limits of the scope is also specifically disclosed. The intermediate value in any stated value or stated range, and each smaller range between any other stated value or intermediate value in the described range is also included in the present invention. The upper and lower limits of these smaller ranges can be independently included or excluded in the scope.

[0035] Unless otherwise indicated, all technical and scientific terms used herein have the same meanings as those generally understood by those skilled in the art. Although the present invention describes only preferred methods and materials, any methods and materials similar or equivalent to those described herein may also be used in the implementation or testing of the present invention. All documents mentioned in this specification are incorporated by reference to disclose and describe the methods and / or materials associated with the documents. In the event of a conflict with any incorporated document, the content of this specification shall prevail.

[0036] It will be apparent to those skilled in the art that various modifications and variations may be made to the specific embodiments of the present invention description without departing from the scope or spirit of the present invention. Other embodiments derived from the present invention description will be apparent to those skilled in the art. The present invention description and examples are exemplary only.

[0037] The words “include,” “including,” “have,” “contain,” etc. used in this document are open-ended terms, meaning including but not limited to.

[0038] The present invention first screened the OsDEL1 gene that responds to drought stress from the wild type rice ZH11 (Zhonghua 11, conventional purchase) by transcriptome sequencing analysis, indicating that OsDEL1 may be involved in regulating the drought resistance of rice. In order to verify this hypothesis, the present invention used real-time fluorescence quantitative PCR to detect the expression of OsDEL1 in wild type rice ZH11 after 20% PEG treatment (simulating drought stress), and found that the expression of OsDEL1 was significantly downregulated after 20% PEG treatment for 0.5h-24h, indicating that OsDEL1 can respond to drought stress.

[0039] Example 1 Real-time fluorescence quantitative PCR analysis of OsDEL1 response to drought

[0040] 1. Primer Design for Real-time Quantitative PCR

[0041] Nucleotide sequence of OsDEL1:

[0042]

[0043] Amino acid sequence of the protein encoded by OsDEL1:

[0044] MATAAVMAAVPSSSPADAAEAVVMTEAVPSLPQRQQPVFVEGRGGKLRDHAYSRKQKSLGLLCSNFVALYNRDDVESIGLDDAARRLGVERRRIYDIVNVLESVGILVRKA KNRYSWIGFGGVPMALRELKERALREKSGLAPLPVEEPSAAIMSDDEDEDKMGDADGDTESEKLSQPVDNPSDNKPGAPRCRLRSDHRKEKSLGLLTQNFVKLFLTMEVDTI SLDEAAKLLLGEGHAENSMRTKVRRLYDIANVLSSLNFIDKIQQADSRKPAFRWLGSAGKPKAENGVTIAVPPPGKTISNKRAFGTELTNIDINRSRLDSTIPKKAKLTLS GGEILKNCKLSVQKQLGQGSKGGFVYGPFHPAGARKQELDNGNKGHTDNVQNWESLAASFRPQYQNQALGDLFAHYVEAWKSWYSEFAQGSSMMQQHFGMPVINQFL*(SEQ IDNO.2).

[0045] The following real-time quantitative PCR primers were designed based on the nucleotide sequence of OsDEL1:

[0046] OsDEL1-qF (upstream primer): GACTCAACAATCCCAAAGAA (SEQ ID NO. 3);

[0047] OsDEL1-qR (downstream primer): TTTCCAGGCTTCCACATA (SEQ ID NO. 4).

[0048] 2. Material cultivation and stress treatment

[0049] (1) The experimental rice material was wild type rice ZH11, which was cultured in hydroponic boxes until the four-leaf stage.

[0050] (2) Leaves of rice seedlings were collected at different time points, such as 0.5 h, 2 h, 6 h, 8 h, 12 h and 24 h after treatment with 20% PEG, and 2 h, 6 h and 12 h after treatment with 50 μm MABA. They were quickly frozen in liquid nitrogen and kept in a -80 °C refrigerator. Total RNA of rice was extracted using Trizol reagent, and RNA was reverse transcribed into cDNA. cDNA was used as a template for real-time fluorescence quantitative PCR (real-time fluorescence quantitative PCR reaction system and procedure: refer to Thermo Fisher Scientific Prepare the reaction system according to the instructions of Green qPCR SuperMix (operate on ice). Use two-step PCR reaction and the standard amplification procedure is as follows:

[0051]

[0052] The results are as follows Figure 1 As shown in the figure, the expression of OsDEL1 gene in ZH11 was significantly downregulated after treatment with 20% PEG and 50 μM ABA, indicating that this gene plays an important role in drought stress.

[0053] Example 2 Construction of OsDEL1 overexpression and knockout transgenic rice

[0054] 1. The PCAMBIA1301 vector (purchased from Newpro Biotech) was used to construct an OsDEL1 overexpression vector, and the 35S promoter was used to drive the expression of OsDEL1, as follows:

[0055] (1) Based on the nucleotide sequence of OsDEL1, the following primers were designed to amplify its CDS sequence:

[0056] OsDEL1-ox-F: TGACCATGGTAGATCTCTATGGCGACGGCGGCGGT (SEQ ID NO.5);

[0057] OsDEL1-ox-R: AATTCGAGCTGGTGACCCAAAAACTGGTTAATGACAGGCA (SEQ ID NO. 6).

[0058] (2) Total RNA was extracted from leaves of wild-type ZH11 grown under normal conditions and reverse transcribed to obtain cDNA. Using cDNA as a template, a high-fidelity DNA polymerase Max Master Mix was used to amplify the rice OsDEL1 gene. The PCR product was detected by 1% agarose gel electrophoresis, and a single DNA band (1321 bp) was obtained, which was the OsDEL1 gene fragment obtained by PCR amplification. The OsDEL1 fragment was recovered by agarose gel electrophoresis according to the instructions of Magen's HiPure Gel Pure DNA Kits.

[0059] The PCAMBIA1301 vector was double-digested with BglII and BstEII and the digestion products were recovered. According to the instructions of ClonExpress II One Step Cloning Kit, the OsDEL1 gene fragment recovery product was connected to the PCAMBIA1301 vector using the homologous recombination method and the connection system in Table 1 to obtain the OsDEL1 overexpression vector (OsDEL1-ox).

[0060] Table 1 Connection system

[0061]

[0062]

[0063] Note: X / Y is calculated according to the formula to obtain the amount of vector and insert (calculation formula: optimal amount of cloning vector = [0.02 × number of base pairs of cloning vector] ng; optimal amount of insert = [0.04 × number of base pairs of insert] ng). In Table 1, X is 4 μL and Y is 10 μL.

[0064] 2. Use pYLCRISPR / Cas9Pubi-H vector to construct the knockout vector of OsDEL1, as follows:

[0065] (1) Based on the nucleotide sequence of OsDEL1, the following target sequences were selected on the first exon and the second exon:

[0066] Target 1: TCGTCTTCGCCGGCCGACGC (SEQ ID NO. 7);

[0067] Target 2: GCGACGACGTGGAGTCTATC (SEQ ID NO. 8);

[0068] The target primers were synthesized according to the target sequence as follows:

[0069] OsDEL1-gRT1: CGTCGGCCGGCGAAGACGAGTTTTAGAGCTAGAAAT (SEQ ID NO. 9);

[0070] OsDEL1-OsU6aT1: TCGTCTTCGCCGGCCGACGGGCAGCCAAGCCAGCA (SEQ ID NO. 10);

[0071] OsDEL1-gRT2: CGACGACGTGGAGTCTATCGTTTTAGAGCTAGAAAT (SEQ ID NO. 11);

[0072] OsDEL1-OsU6aT2: GATAGACTCCACGTCGTCGGGCAGCCAAGCCAGCA (SEQ ID NO. 12);

[0073] (2) Construction of sgRNA expression cassette (taking target 1 as an example):

[0074] 1) Take 2-5 ng pYLgRNA-OsU6a plasmid as template, primers UF and OsDEL1-OsU6aT1 0.2 μM each, gR-R and OsDEL1-gRT1 0.1 μM each, and use high-fidelity enzyme Max Master Mix was used for the first round of PCR reaction. The PCR reaction procedure was as follows: 25-28 cycles: 94°C for 10 s, 58°C for 15 s, and 72°C for 20 s. During the amplification process, overlapping PCR was used to generate the sgRNA expression cassette fragment of the two fragments. The primer UF sequence is shown in SEQ ID NO.13, and the primer gR-R sequence is shown in SEQ ID NO.14;

[0075] UF: CTCCGTTTTACCTGTGGAATCG (SEQ ID NO.13);

[0076] gR-R: CGGAGGAAAATTCCATCCAC (SEQ ID NO. 14).

[0077] 2) Take 1 μl of the first round PCR product and hydrate it with ddH 2 O was diluted 10 times and 1 μl was used as template. Pps-GGL and Pgs-GGR were added as primers (final concentration 0.15 μM). High-fidelity enzyme Max Master Mix amplification, PCR program: 17-20 cycles: 94°C 10s, 58°C 15s, 72°C 20s. PCR products were detected by 1% agarose gel electrophoresis, and the sgRNA expression cassette was recovered. The primer Pps-GGL sequence is shown in SEQ ID NO.15, and the primer Pgs-GGR sequence is shown in SEQ ID NO.16;

[0078] Pps-GGL: TTCAGAGGTCTCTCTCGACTAGTATGGAATCGGCAGCAAAGG (SEQ ID NO. 15);

[0079] Pgs-GGR: AGCGTGGGTCTCGACCGACGCGTATCCATCCACTCCAAGCTC (SEQ ID NO. 16).

[0080] 3) Assemble the sgRNA expression cassette into the pYLCRISPR / Cas9Pubi-H vector. Perform the enzyme digestion-ligation reaction on the pYLCRISPR / Cas9Pubi-H plasmid and the sgRNA expression cassette according to the enzyme digestion-ligation system shown in Table 2, and perform enzyme digestion and ligation using variable temperature cycles: first 3 cycles (37°C 10 min, 10°C 5 min, 20°C 5 min); then 10 cycles (37°C 3 min, 10°C 5 min, 20°C 5 min), and finally 37°C 5 min. The ligation product was obtained;

[0081] Table 2 Enzyme digestion-ligation system

[0082]

[0083] 4) The ligation product in step 3) was transformed into DH5α competent cells by heat stimulation, and positive clones were identified by blue-white colony screening and bacterial liquid PCR detection to obtain the OsDEL1 gene knockout vector.

[0084] Example 3 Obtaining OsDEL1 overexpression and knockout transgenic rice and identifying positive plants

[0085] The overexpression and knockout vector plasmids of OsDEL1 obtained in Example 2 were transferred into Agrobacterium EHA105. The overexpression and knockout vectors were transformed into the wild-type ZH11 variety by the genetic transformation method mediated by Agrobacterium EHA105 to obtain transgenic plants. The hygromycin gene Hpt was detected by PCR, and the expression of OsDEL1 was detected by real-time quantitative PCR to obtain transgenic positive plants OsDEL1-ox-1, OsDEL1-ox-16 and OsDEL1-ox-17 ( Figure 2 ).

[0086] The following method was used to detect the targeting effect of OsDEL1 gene knockout transgenic rice:

[0087] (1) T extracted by CTAB method 0The DNA of the knockout transgenic rice was used to amplify the target sequences of the first exon and the second exon region of the OsDEL1 gene by PCR, and the amplified products were sent to Shanghai Sangon Biotechnology Co., Ltd. for sequencing. The PCR amplification primer sequences for the target sequences of the first exon and the second exon region of the OsDEL1 gene are as follows.

[0088] OsDEL1-csa9-JC-target1-F: GCCAAATCAAAGCCCCCTCT (SEQ ID NO. 17);

[0089] OsDEL1-csa9-JC-target1-R: CCAAAATCCACGAATCCCCC (SEQ ID NO. 18).

[0090] OsDEL1-csa9-JC-target2-F:TTATGACGGAGGCGGTGC (SEQ ID NO. 19);

[0091] OsDEL1-csa9-JC-target2-R: AATGCCATGGGACGCC (SEQ ID NO. 20).

[0092] (2) Compare the sequencing results with the target sequence to determine whether the target site has a single base insertion, deletion, mutation, or a deletion of several bases to determine whether the target site is successful.

[0093] The results are as follows Figure 3 As shown, the target sequence in OsDEL1 gene knockout transgenic rice del1-1 and del1-2 produced a 1 bp base insertion compared with the wild type WT, proving that the target sequence of OsDEL1 gene knockout transgenic rice was successfully targeted.

[0094] Example 4 Identification of Drought Resistance of OsDEL1 Gene Overexpression and Knockout Transgenic Rice Plants

[0095] The drought resistance of the positive offspring of the overexpression and knockout transgenic rice obtained in Example 3 was tested. The specific testing method is as follows:

[0096] The OsDEL1-ox, del1 transgenic rice and ZH11 seedlings that had been hydroponically cultured in the conventional rice nutrient solution of the International Rice Research Institute for about 15 days were dehydrated for 24 hours to induce water shortage stress. Then a rehydration experiment was conducted. After 6-7 days of rehydration, photos were taken and the survival rate was calculated ( Figure 4-Figure 6 ). The results showed that the survival rate of transgenic rice with overexpression of OsDEL1 gene under drought stress was lower and its drought resistance was weaker; while the survival rate of del1 transgenic rice under drought stress was higher and its drought resistance was stronger.

[0097] Example 5 Measurement of water loss rate of OsDEL1 gene transgenic rice plants

[0098] The leaves of ZH11 and OsDEL1 gene overexpression and knockout transgenic rice plants at the heading stage were cut into 2 cm lengths and divided into three equal parts, with 3 replicates. They were placed in a culture dish and naturally dehydrated at room temperature (25°C). The weight of the leaves was measured at 0, 1, 2, 4, 6, 18, and 20 hours, and the dehydration rate was calculated as (weight before dehydration - weight after dehydration) / weight before dehydration. "ZH11" represents the wild type, and * represents a significant difference compared with the wild type (*, P<0.05, **, P<0.01, ***, P<0.001). The statistical method used was the T-test test. Figure 7 The results showed that the water loss rate of OsDEL1 overexpressed transgenic rice was significantly higher than that of ZH11, and the water loss rate of OsDEL1 knockout transgenic rice was significantly lower than that of ZH11. This indicates that knocking out the OsDEL1 gene can improve rice drought resistance by reducing the water loss rate.

[0099] Example 6 Experiment on ABA sensitivity of OsDEL1 gene knockout transgenic rice plant del1

[0100] The del1 transgenic rice and wild-type ZH11 seeds were aseptically sown in 1 / 2MS medium with ABA concentrations of 0 μM and 5 μM, respectively, and germinated at 28°C under a 15 / 24 h photoperiod. Phenotypic analysis and photography were performed 10 days after treatment, and root length and shoot length ( Figure 8 and Fig. 9 ), the results showed that del1 transgenic rice seeds were more sensitive to ABA during germination, indicating that OsDEL1 may affect rice drought resistance through the ABA signal transduction pathway.

[0101] The embodiments described above are only descriptions of the preferred modes of the present invention, and are not intended to limit the scope of the present invention. Without departing from the design spirit of the present invention, various modifications and improvements made to the technical solutions of the present invention by ordinary technicians in this field should all fall within the protection scope determined by the claims of the present invention.

Claims

1. A drought resistance gene OsDEL1, characterized in that: The nucleotide sequence of the drought resistance gene OsDEL1 is shown in SEQ ID NO.

1.

2. The protein encoded by the drought resistance gene OsDEL1 according to claim 1.

3. The protein according to claim 2, characterized in that The amino acid sequence of the protein is shown in SEQ ID NO.

2.

4. A set of primer pairs for amplifying the drought resistance gene OsDEL1 according to claim 1, characterized in that: It includes an upstream primer with a nucleotide sequence such as SEQ ID NO.3 and a downstream primer with a nucleotide sequence such as SEQ ID NO.

4.

5. A method for amplifying the drought resistance gene OsDEL1 according to claim 1 using the primer pair according to claim 4, characterized in that: The following steps are involved: The plant nucleic acid molecule is used as a template, the template and a primer pair are prepared into a PCR amplification system for PCR amplification to obtain the drought resistance gene OsDEL1.

6. A method for improving drought resistance of plants, characterized in that: By reducing the expression level of the OsDEL1 gene according to claim 1, the drought resistance of plants is improved.

7. Use of the drought-resistant gene OsDEL1 according to claim 1, the protein according to claim 2 or 3, or the drought-resistant gene OsDEL1 amplified by the method according to claim 5 in improving plant drought resistance.

8. The use according to claim 7, characterized in that: The plants include rice.

9. Use of the drought-resistant gene OsDEL1 according to claim 1, the protein according to claim 2 or 3, or the drought-resistant gene OsDEL1 amplified by the method according to claim 5 in cultivating different drought-resistant crop germplasms.

10. The use according to claim 9, characterized in that: The drought resistance is reduced by overexpressing the expression level of the drought resistance gene OsDEL1; and the drought resistance is enhanced by reducing the expression level of the drought resistance gene OsDEL1.

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