CN109536509B

CN109536509B - Sesame drought-resistant, moisture-resistant and salt-tolerant gene SiNAC56, protein coded by same and application of sesame drought-resistant, moisture-resistant and salt-tolerant gene SiNAC56

Info

Publication number: CN109536509B
Application number: CN201811408071.XA
Authority: CN
Inventors: 黎冬华; 山姆; 游均; 玛丽; 周瑢; 刘爱丽; 张秀荣
Original assignee: Oil Crops Research Institute of Chinese Academy of Agriculture Sciences
Current assignee: Oil Crops Research Institute of Chinese Academy of Agriculture Sciences
Priority date: 2018-11-23
Filing date: 2018-11-23
Publication date: 2021-11-23
Anticipated expiration: 2038-11-23
Also published as: CN109536509A

Abstract

The invention belongs to the technical field of genetic engineering, and provides a sesame drought-resistant, moisture-resistant and salt-tolerant gene SiNAC56, and a protein coded by the same and application of the sesame drought-resistant, moisture-resistant and salt-tolerant gene SiNAC 56; the sesame drought-resistant, moisture-resistant and salt-tolerant gene SiNAC56 is a nucleotide sequence shown in SEQ ID NO. 1; or a nucleotide sequence with drought resistance generated by adding, substituting, inserting or deleting one or more nucleotides in the nucleotide sequence shown in SEQ ID NO. 1; the invention constructs the SiNAC56 gene into an expression vector, and transfers the SiNAC56 gene into arabidopsis thaliana by an agrobacterium-mediated transformation method to obtain transgenic arabidopsis thaliana; through the overexpression of the sesame gene SiNAC56 related to drought resistance, moisture resistance and salt tolerance in arabidopsis thaliana, the gene is found to be capable of remarkably improving the drought resistance, moisture resistance and salt tolerance of plants, so that the invention has a good application prospect in improving the drought resistance, moisture resistance, salt tolerance and other properties of the plants.

Description

Sesame drought-resistant, moisture-resistant and salt-tolerant gene SiNAC56, protein coded by same and application of sesame drought-resistant, moisture-resistant and salt-tolerant gene SiNAC56

Technical Field

The invention relates to the technical field of genetic engineering, in particular to a sesame drought-resistant, moisture-resistant and salt-tolerant gene SiNAC56, and a protein coded by the same and application thereof.

Background

Sesame (Sesamum indicum L.) is also known as an orphan crop. Nowadays, the demand for its seeds is increasing worldwide due to its high oil content (around 50%), protein (around 25%) and rich antioxidant substances (sesamin, lignan, etc.). In addition to these nutrients, sesame planting has many agricultural advantages: it grows well in tropical and temperate climates, does not require rainfall or irrigation, can grow on stored soil moisture, can grow in pure forests with low input, or can grow in mixed forests of multiple crops. Despite the many advantages of sesame, sesame has been studied less, and thus, the work for genetic and breeding improvement of sesame has been greatly limited. The main reason for the limited sesame production is that sesame is mainly distributed in developing countries, usually in small farmers. Although sesame has a high yield potential, the actual yield is rather low and unstable due to the combined action of biotic and abiotic stress, which is also a significant bottleneck problem for the development of the sesame industry.

Sesame is mainly planted in arid and semiarid regions, is easy to suffer from extreme and intermittent drought stress, is difficult to obtain stable and high yield, and has great influence on the quality of the sesame due to the drought stress. In recent years, the drought damage of sesame production in China is increased continuously, and the yield is reduced seriously. The main sesame production areas in Huang-Huai and Changjiang river watersheds are easy to be seasonally arid, and seriously affect the growth and the yield of the sesame. The spring sowing areas of sesame in the west and the northeast of China and the south and autumn sowing areas of Yangtze river suffer from drought threats all year round, and the yield is influenced. The moisture damage is a main abiotic adversity factor influencing the sesame yield stability and improvement of the main producing area of Jianghuai sesame in China, the sesame in the main producing area is likely to suffer from moisture damage stress in the whole growing period (usually 6 months to 9 months), particularly, the seedling stage is just in rainy seasons in most areas, and the moisture damage stress is also usually accompanied with the occurrence of diseases, so that the yield of the sesame is reduced, and even the sesame is completely harvested. While sesame production in western regions and coastal regions of China is mainly influenced by soil salinization, salt tolerance breeding is a decisive factor for improving sesame yield in the regions. Therefore, high and stable yield is obtained under the abiotic stress environment, related resistance genotypes are needed, high and stable yield is promoted by improving drought resistance, moisture resistance and salt tolerance of the sesame, and the method is necessary for promoting the development of the sesame industry in China and solving the problem of insufficient total self-supply. Sesame abiotic stress is weak in research, and molecular related research is less. The development of the functional genes related to abiotic stress resistance of the sesame is an important basis and reliable way for developing the breeding of related sesame resistance molecules so as to improve the abiotic stress resistance of the sesame.

Therefore, the development of a drought-resistant, moisture-resistant and salt-tolerant sesame gene is urgently needed, so that the sesame drought-resistant, moisture-resistant and salt-tolerant molecular breeding is developed and is used as one of important bases and reliable ways for improving the drought-resistant, moisture-resistant and salt-tolerant capacities of the sesame.

Disclosure of Invention

The invention aims to overcome the defects of the prior art and provides a sesame drought-resistant, moisture-resistant and salt-tolerant gene SiNAC56, and a protein coded by the same and application of the gene SiNAC56, wherein the gene SiNAC56 can be used for improving the drought resistance, moisture resistance and salt tolerance of plants so as to be used for breeding drought-resistant, moisture-resistant and salt-tolerant varieties of oil crops, so that the drought resistance, moisture resistance and salt tolerance of the crops are improved, and the high yield and stable yield of the crops are ensured.

To achieve the above object, the present invention is realized by:

in the first aspect of the invention, a drought-resistant, moisture-resistant and salt-tolerant sesame gene SiNAC56 is provided, wherein the drought-resistant, moisture-resistant and salt-tolerant sesame gene SiNAC56 is a nucleotide sequence shown as SEQ ID NO. 1; or a nucleotide sequence with drought resistance, moisture resistance and salt resistance, which is generated by adding, substituting, inserting or deleting one or more nucleotides in the nucleotide sequence shown in SEQ ID NO. 1.

The sesame drought-resistant, moisture-resistant and salt-tolerant gene SiNAC56 is obtained by sequencing based on the whole sesame genome and performing a large amount of bioinformatics analysis and screening by the applicant. The applicant names the gene SiNAC56 for resisting drought, humidity and salt of sesame.

In a second aspect of the invention, the sesame SiNAC56 protein encoded by the sesame drought-resistant, moisture-resistant and salt-tolerant gene SiNAC56 is provided, and the protein is as shown in SEQ ID NO: 2; or an amino acid sequence which has at least 90 percent of homology with the amino acid sequence shown in SEQ ID NO.2 and has drought resistance, moisture resistance and salt resistance.

In a third aspect of the invention, a cloning method of the drought-resistant, moisture-resistant and salt-tolerant sesame gene SiNAC56 is provided, and the method comprises the following steps: extracting sesame RNA, carrying out reverse transcription on the sesame RNA to obtain cDNA, taking the cDNA as a template, and taking the following sequence as a primer, and obtaining a SiNAC56 gene sequence by RT-PCR, wherein the primer sequence is as follows: SiNAC 56-F: SEQ ID NO. 3; SiNAC 56-R: shown as SEQ ID NO. 4.

In the fourth aspect of the invention, an expression vector of the drought-resistant, moisture-resistant and salt-tolerant sesame gene SiNAC56 is provided.

The invention specifically provides an expression vector, wherein the expression vector constructs a pCAMBIA1301S-SiNAC56 vector through a pCAMBIA1301S vector.

In a fifth aspect of the present invention, there is provided a transformant of the expression vector.

In some embodiments, the transformant is an agrobacterium tumefaciens, and/or a plant cell (or organism); the organism is a transgenic drought-resistant plant, and is one of rice, corn, wheat, barley, tobacco, soybean, sorghum, cotton, hemp, peanut, rape, sesame, sugarcane and beet, wherein sesame is preferred.

In the sixth aspect of the invention, a method for improving drought resistance, moisture resistance and salt tolerance of plants is provided, wherein the pCAMBIA1301S-SiNAC56 vector mediates genetic transformation of plants by means of genetic engineering to obtain transgenic plants.

In a seventh aspect of the invention, application of sesame drought resistance, moisture resistance and salt tolerance gene SiNAC56 in improvement of drought resistance, moisture resistance and salt tolerance of plants is provided.

In the eighth aspect of the invention, the application of the expression vector in improving the drought resistance, the moisture resistance and the salt tolerance of plants is provided.

In the ninth aspect of the invention, the application of the transformant in improving the drought resistance, the moisture resistance and the salt tolerance of plants is provided.

The invention constructs the SiNAC56 gene into an expression pCAMBIA1301S vector (pCAMBIA1301S is reconstructed on the basis of a plant genetic transformation vector pCAMBIA1301 commonly used internationally and carries a genetic transformation vector of a cauliflower mosaic virus CaMV35S promoter with constitutive and over-expression characteristics). The SiNAC56 gene carried by pCAMBIA1301S is transferred into arabidopsis thaliana by an agrobacterium-mediated transformation method to obtain an arabidopsis thaliana transformation plant. Experimental results show that the SiNAC56 has the function of improving drought resistance, moisture resistance and salt tolerance of plants.

The invention has the beneficial effects that:

the sesame drought-resistant, moisture-resistant and salt-tolerant gene SiNAC56 provided by the invention is firstly reported at home and abroad, the SiNAC56 gene is constructed into an expression pCAMBIA1301S vector, and the SiNAC56 gene carried by pCAMBIA1301S is transferred into arabidopsis thaliana by an agrobacterium-mediated transformation method to obtain transgenic arabidopsis thaliana; through the overexpression of the sesame gene SiNAC56 related to drought resistance, moisture resistance and salt tolerance in arabidopsis thaliana, the gene is found to be capable of remarkably improving the drought resistance, moisture resistance and salt tolerance of plants. Therefore, the overexpression of the drought-resistant, moisture-resistant and salt-tolerant gene SiNAC56 of the sesame can be used for improving the drought-resistant, moisture-resistant and salt-tolerant capability of plants so as to be used for breeding drought-resistant varieties of oil crops, and therefore the high and stable yield of the crops is guaranteed.

Drawings

FIG. 1 shows the construction of a plant expression vector pCAMBIA1301S-SiNAC56 containing SiNAC56 gene;

FIG. 2 shows the cleavage map of vector pCAMBIA1301S-SiNAC56 (M1: 1kb DNA ladder; M2: 2000DNA Marker; 1-2: pCAMBIA1301S-SiNAC56 cleavage fragment);

FIG. 3 is a schematic diagram showing the screening results of positive plants of the T1 generation of Arabidopsis thaliana transformed with SiNAC56 gene;

FIG. 4 is a schematic diagram showing PCR identification results of T1 generations of SiNAC56 transgenic Arabidopsis thaliana (M: Marker; 1-6: transgenic T1 plants; WT: wild type Arabidopsis thaliana);

FIG. 5 is a schematic diagram showing the screening results of positive plants of the T2 generation of Arabidopsis thaliana transformed with SiNAC56 gene;

FIG. 6 is a schematic diagram showing PCR identification results of T2 generations of SiNAC56 transgenic Arabidopsis thaliana (M: Marker; 1-9: transgenic T2 plants);

FIG. 7 is a diagram showing the result of verification of quantitative expression of T2 generation plants of SiNAC56 transgenic Arabidopsis;

FIG. 8 is a soil drying method for determining drought resistance of Arabidopsis T2 generation plants transformed with SiNAC56 gene;

FIG. 9 shows moisture resistance of SiNAC56 transgenic Arabidopsis T2 plant measured by pot flooding method;

FIG. 10 shows the salt tolerance of Arabidopsis T2 generation plants with SiNAC56 gene transferred by NaCl salt stress method.

Detailed Description

Example 1 obtaining of drought, moisture and salt resistant Gene SiNAC56 Gene of sesame

Firstly, discovery of drought-resistant, moisture-resistant and salt-tolerant sesame gene SiNAC56

1. A pair of sesame drought-resistant materials ZZM0635 and drought-sensitive materials ZZM4728 are selected, potted and planted in a drought shed, and drought stress treatment is carried out in the initial flowering period (47 d after sowing). Taking 2 leaves of materials at 5 time points of d0 (soil moisture content 35%), d1 (soil moisture content 15%), d2 (soil moisture content 9%), d3 (soil moisture content 6%) and d4 (soil moisture content 35% after rehydration) respectively for drought-resistant transcriptome analysis;

2. using the Illumina Hiseq4000 sequencing platform, transcriptome sequencing was performed on 30 samples, yielding an average of 6.86Gb data per sample. After aligning sequencing reads to sesame reference genome and reconstructing transcripts, finally detecting 24218 new transcripts in total, wherein 20236 transcripts belong to the new alternative splicing subtype of the known protein coding gene, 1806 transcripts belong to the new protein coding gene, and the remaining 2176 transcripts belong to long-chain non-coding RNA;

3. through differential expression gene analysis, 682 Differential Expression Genes (DEGs) are found in the drought-resistant material, and the genes comprise the functions of interleukin related functional receptor related kinase, cytochrome P450, HSP20 family protein, glutathione S transferase, superoxide dismutase and the like. Through analysis of genes involved in transcription regulation, 2164 transcription factors belonging to 46 families are found to participate in drought response, wherein MYB family members are the most abundant and active transcription factors, and are transcription factors such as AP2-EREBP, bHLH, WRKY, C2H2, bZIP, GRAS, NAC and the like;

4. through sesame reference genome (http:// ocri-genomics.org/Sinbase/index. html) comparison and gene annotation analysis and quantitative expression verification (drought resistance, moisture resistance and salt tolerance), 1 NAC transcription factor gene SIN _1026079 is found to have very high expression level under 3 adverse conditions of drought, moisture and salt stress, the Arabidopsis homologous gene of the gene is ANAC104, is a transcription factor gene involved in plant growth and development and is related to plant adverse stress regulation, so that the gene is presumed to be possibly related to drought resistance sesame, moisture resistance and salt tolerance, and is named SiNAC 56.

Secondly, extracting RNA of sesame root system and cloning of drought-resistant, moisture-resistant and salt-tolerant sesame gene SiNAC56

1. The extraction of root system RNA adopts an improved CTAB method, and the synthesis of cDNA is carried out according to the procedures of a reverse transcription synthesis kit of TaKaRa company. Primers capable of amplifying the complete CDS sequence of the SiNAC56 gene were designed using Primer5.0, based on the CDS sequence of the SiNAC56 gene, and the sequences (including modified bases) and names were as follows:

SiNAC 56-F: 5'-GCTTTCGCGAGCTCGGTACCATGGCTGAAGGGAGGAAATG-3' (shown as SEQ ID NO. 3)

SiNAC 56-R: 5'-AGATCAGCTTGCCTAACTAGCTAGTTAGGCAAGCTGATCT-3' (shown in SEQ ID NO. 4);

2. taking leaves of the early flowering stage drought resistant germplasm ZZM1805 subjected to drought stress for 5 days, extracting total RNA of a root system, carrying out reverse transcription to generate cDNA, taking the reversed cDNA as a template, carrying out RT-PCR amplification by using a primer SiNAC56-F/R in the step (5), and sequencing the amplified fragments to obtain a SiNAC56 gene sequence for improving the drought resistance of sesame, wherein the SiNAC56 gene sequence is shown in SEQ ID NO. 1.

Example 2 construction and genetic transformation of SiNAC56 Gene expression vector

First, construction of overexpression vector

The sesame gene SiNAC56 related to drought resistance, moisture resistance and salt tolerance cloned in the embodiment 1 is connected with pCAMBIA1301S (provided by the laboratory) plasmid by utilizing a homologous recombination method to construct a plant expression vector, which is named as pCAMBIA1301S-SiNAC56 (shown in figure 1), and the specific operation is as follows:

1. firstly, a linearized vector is obtained by a double enzyme digestion (BamHI and KpnI) (Takara) method, and then the linearized vector with high purity is obtained by agarose gel electrophoresis and purification with a gel recovery kit (Tiangen Biochemical technology Co., Ltd.).

2. Adding the target fragment DNA and the linearized vector into a 1.5ml centrifuge tube in a molar ratio of 3:1 for recombination reaction, uniformly mixing, standing at 37 ℃ for about 30min, adding 10 ul of reaction solution into 50 ul of DH5a competent cells, gently mixing by using a pipette, incubating on ice for 20min, performing heat shock in a 42 ℃ water bath for 45 seconds, and rapidly cooling on ice for 2 min.

3. Adding 300. mu.l LB liquid medium, and incubating at 37 ℃ for 45-60 min. Centrifuging at 5,000rpm for 2min, collecting thallus, discarding part of supernatant, re-suspending thallus with the rest culture medium, lightly spreading on LB solid culture medium containing Kan resistance with sterile spreading rod, and culturing at 37 deg.C for 16-24 hr by inversion in incubator.

4. Selecting a plurality of clones on the recombinant reaction conversion plate to carry out colony PCR identification, identifying as positive colonies, selecting corresponding single colonies to culture in a liquid LB culture medium containing Kan antibiotics at 37 ℃ and 200rpm for overnight, extracting plasmids or directly sequencing bacterial liquid, and identifying the carrier accuracy through enzyme digestion electrophoresis (figure 2).

Second, genetic transformation

Transferring the vector pCAMBIA1301S-SiNAC56 prepared in the step 1) into Agrobacterium tumefaciens LBA4404 (Shanghai Weidi Biotechnology Co., Ltd.), and then introducing into an Arabidopsis plant, wherein the specific operations are as follows:

1. the recombinant vector is transferred into agrobacterium LBA 4404:

(1) add 2. mu.g plasmid DNA into every 100. mu.l LBA4404 Agrobacterium tumefaciens competent cells, dial the tube bottom by hand and mix evenly, and stand on ice for 5min, liquid nitrogen for 5min, water bath at 37 ℃ for 5min, ice bath for 5min in turn.

(2) Adding 700 μ l LB liquid culture medium without antibiotics, and shake culturing at 28 deg.C for 5 h; and (3) centrifuging at 6000rpm for 1min to collect thalli, reserving about 100 mu l of supernatant, slightly blowing and beating the resuspended thalli, uniformly coating the thalli on an LB solid culture medium containing Kan and Rif, inversely placing the thalli in an incubator at 28 ℃ for 2 days, and picking a plurality of positive clones to simply verify the result by utilizing colony PCR.

2. Plate culture of Arabidopsis thaliana:

(1) a certain amount of arabidopsis seeds are counted according to the experiment requirement and are filled in a sterile 1.5mL centrifuge tube.

(2) 1mL of 75% ethanol was added, the mixture was inverted and mixed, and the supernatant was discarded and repeated 1 time. Placing into a shaker at 37 deg.C and 200rpm, and shaking for 10min for surface sterilization.

(3) Discard 75% ethanol, add 1mL 95% ethanol, mix by inversion, discard the supernatant, and repeat 1 time. In a clean bench, 300-.

(4) After the ethanol is volatilized, the arabidopsis seeds are dibbled on a prepared plate by using toothpicks.

(5) Sealing the flat plate, reversely placing the flat plate at 4 ℃ under the dark condition for vernalization for 48h, placing the flat plate in an illumination incubator for vertical culture after vernalization is finished, and transplanting after seedling emergence for one week.

(6) The seedlings were planted in soil of a small pot with tweezers, first kept wet for 24h with a preservative film, placed in the plant growth room and cultured until the growth of Arabidopsis thaliana bolting (about one month) for transformation experiments.

3. Genetic transformation:

(1) activating agrobacterium: 20 mu.L of Rif and Kan are added into 20mL of LB liquid culture medium respectively, shaken and inoculated, and activated by shaking at 220rpm at 28 ℃ for 8-10 h.

(2) And (3) agrobacterium tumefaciens enlarged culture: respectively adding 200 mul of Rif and Kan into 200mL of LB liquid culture medium, adding 5-10mL of activated bacterium liquid, shake culturing at 28 ℃ and 220rpm for 14-16h until OD value is 1.6-2.0, centrifuging at 4500rpm for 10min, removing supernatant from the precipitated thallus, and naturally drying.

(3) And adding 100mL of 5% sucrose solution into the precipitated bacteria to resuspend the bacteria, and blowing and beating the bacteria uniformly by a pipette to resuspend the bacteria.

(4) Adding the bacterial liquid in the centrifuge flask into a plate, adding 100mL of 5% sucrose solution, adding 40 μ L of Silwet-L-77 (0.02%) before conversion, shaking the plate, and mixing uniformly.

(5) The Arabidopsis inflorescences were closed, immersed in a plate, and gently shaken for 15 s. After the conversion, the bacterial liquid is stirred evenly.

(6) The plants were covered with a black bag, protected from light and kept moist for 24 h.

(7) The transformation was repeated once more after one week.

Thirdly, screening and identifying over-expression plants

1. Screening positive plants of T1 generation.

Seeds harvested from T0 generation of arabidopsis thaliana are planted, seeds of T0 generation are disinfected, MS screening culture medium containing 30mg/L hygromycin (25 mg/L of cefamycin is added for bacteriostasis) is inoculated for illumination culture for 7-10 days at 22 ℃, positive plants (plants with normal growth of seedlings and roots) are obtained through screening (figure 3), 6 positive strains are obtained through the experiment, the positive seedlings are transplanted into soil, the positive seedlings are covered with preservative film for 2-3 days, then the film is uncovered, and then the plants grow normally. After the DNA of the leaves of the screened positive plants is extracted, the SiNAC56 gene is identified by a PCR method, the molecular verification of the target gene of the transgenic plants is carried out (figure 4), and finally the gene is confirmed to be transferred into T1 generation positive plants.

2. Positive detection of transgenic plant T2 generation

And (3) performing single plant seed collection on the T1 generation positive plants to obtain T1 generation seeds, continuously performing hygromycin screening to obtain T2 generation positive plants (figure 5), transplanting the obtained positive plants to grow, extracting leaf genome DNA (shown in figure 6) to perform PCR molecular identification, and determining the T2 generation positive plants.

3. Quantitative expression verification of transgenic T2 positive plants

A transgenic plant T2 generation positive plant and a young leaf of a wild type arabidopsis plant in the growth period are taken, total RNA of the leaf is extracted by an RNA extraction kit (Beijing Adela biotechnology limited), then cDNA is obtained by a reverse transcription kit (Nanjing NuoZan biotechnology limited), respective cDNA is taken as a template, arabidopsis beta-actin is taken as an internal reference, and qRT-PCR expression verification (qRT-PCR Mix: Nanjing NuoZan biotechnology limited; instrument: Roche LightCyclerR480) is carried out.

The qRT-PCR primer pair of the arabidopsis beta-actin internal reference comprises the following components:

aF: 5'-CCCGCTATGTATGTCGCCA-3' (shown in SEQ ID NO. 5);

aR: 5'-AACCCTCGTAGATTGGCACAG-3' (shown in SEQ ID NO. 6),

the target gene quantitative qRT-PCR primer pair is as follows:

NACF (NACF): 5'-TCATCGTGGTCGTTGTCG-3' (shown in SEQ ID NO. 7);

NACR: 5'-ATGGCTGAAGGGAGGAAA-3' (shown in SEQ ID NO. 8).

The result shows that the sesame SiNAC56 gene is significantly improved in expression level in the detected leaves of 3 Arabidopsis transgenic lines by using non-transgenic wild Arabidopsis as a control, and the expression of the sesame SiNAC56 gene is not detected in the leaves of the non-transgenic control Arabidopsis (FIG. 7). The SiNAC56 genes of sesame are transformed, inserted into the corresponding arabidopsis genome and expressed.

The 3 SiNAC56 overexpression transgenic T2 generation positive plants are subjected to drought resistance, moisture resistance, salt tolerance and other performances determination

First, determination of drought resistance of T2 generation positive Arabidopsis plants

When the transgenic plants of the T2 generation grow to 3 pairs of leaves, drought stress treatment is respectively carried out on the arabidopsis transgenic plants and the non-transgenic wild arabidopsis plants for 17 days. The result shows that compared with the wild type arabidopsis thaliana control, the drought resistance of the SiNAC56 transgenic arabidopsis thaliana strain obtained in the research is obviously higher than that of the wild type arabidopsis thaliana control (figure 8), and the overexpression of the sesame SiNAC56 gene can improve the drought resistance of the plant.

Second, T2 generation positive Arabidopsis thaliana plant moisture resistance assay

When the transgenic plants of the T2 generation grow to 3 pairs of leaves, the transgenic plants of arabidopsis thaliana and the non-transgenic wild arabidopsis thaliana plants are respectively treated by the wet damage stress (the water submerges 1cm in the soil) for 11 days. The result shows that compared with the wild type arabidopsis thaliana control, the moisture resistance of the SiNAC56 transgenic arabidopsis thaliana strain obtained in the research is obviously higher than that of the wild type arabidopsis thaliana control (figure 9), and the moisture resistance of the plant can be improved by overexpression of the sesame SiNAC56 gene.

Third, T2 generation positive Arabidopsis thaliana plant salt tolerance determination

When transgenic plants of T2 generation grew to 3 pairs of leaves, salt stress (1 time pouring 30ml of 200mM saline every 2 days) was applied to Arabidopsis transgenic plants and non-transgenic wild type Arabidopsis plants for 7 days. The results show that compared with wild type arabidopsis thaliana contrast, the salt tolerance of the SiNAC56 transgenic arabidopsis thaliana strain obtained in the research is obviously higher than that of the wild type arabidopsis thaliana contrast (figure 10), and the salt tolerance of the plant can be improved by overexpression of sesame SiNAC56 gene.

The above description is only for the purpose of illustrating the preferred embodiments of the present invention and is not to be construed as limiting the invention, and any modifications, equivalents, improvements and the like that fall within the spirit and principle of the present invention are intended to be included therein.

Sequence listing

<110> institute of oil crop of academy of agricultural sciences of China

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atggctgaag ggaggaaatg gtatttttac agcaggagga ctcaaagtag ggttactgag 60

agtggatatt ggcagtctat aggggttgaa gaaccaatct tttcaagctc tggccaaaga 120

attggtatga agaaatactg tgctttttac attggtgaac cctcggaagg cgtcaagacc 180

aactggatta tgcaggaata tagactcatg gattcaagtt ccaccagtag atcatctaaa 240

aggagaaatt ccaagataga ttatagtaaa tgggtggttt gtctagtgta tgagaaaggc 300

gacaacgacc acgatgaggg gacagagctt tcatgcttgg atgaagtttt tctgtcgatg 360

gacgatctcg atgagatcag cttgcctaac tag 393

<210> 2

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<213> sesame (Sesamum indicum)

<400> 2

Met Ala Glu Gly Arg Lys Trp Tyr Phe Tyr Ser Arg Arg Thr Gln Ser

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20 25 30

Ile Phe Ser Ser Ser Gly Gln Arg Ile Gly Met Lys Lys Tyr Cys Ala

35 40 45

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50 55 60

Gln Glu Tyr Arg Leu Met Asp Ser Ser Ser Thr Ser Arg Ser Ser Lys

65 70 75 80

Arg Arg Asn Ser Lys Ile Asp Tyr Ser Lys Trp Val Val Cys Leu Val

85 90 95

Tyr Glu Lys Gly Asp Asn Asp His Asp Glu Gly Thr Glu Leu Ser Cys

100 105 110

Leu Asp Glu Val Phe Leu Ser Met Asp Asp Leu Asp Glu Ile Ser Leu

115 120 125

Pro Asn

130

<210> 3

<211> 40

<212> DNA

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gctttcgcga gctcggtacc atggctgaag ggaggaaatg 40

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agatcagctt gcctaactag ctagttaggc aagctgatct 40

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<400> 5

cccgctatgt atgtcgcca 19

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aaccctcgta gattggcaca g 21

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tcatcgtggt cgttgtcg 18

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<212> DNA

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atggctgaag ggaggaaa 18

Claims

1. The application of the sesame drought-resistant, moisture-resistant and salt-tolerant gene SiNAC56 and/or an expression vector containing the sesame drought-resistant, moisture-resistant and salt-tolerant gene SiNAC56 in improving the drought resistance, moisture resistance and salt tolerance of plants is characterized in that the sesame drought-resistant, moisture-resistant and salt-tolerant gene SiNAC56 is a nucleotide sequence shown as SEQ ID NO. 1.

2. The use of claim 1, wherein the expression vector is constructed as pCAMBIA1301S-SiNAC56 vector by pCAMBIA1301S vector.

3. A method for improving drought resistance, moisture resistance and salt tolerance of plants is characterized in that the pCAMBIA1301S-SiNAC56 vector in claim 2 is used for mediating plant genetic transformation by means of genetic engineering to obtain transgenic plants, so that the drought resistance, the moisture resistance and the salt tolerance of the plants are improved.