EmSAMS1 protein derived from elaeagnus pungens and application of encoding gene of EmSAMS1 protein in improvement of plant salt tolerance

Abstract

The invention discloses an application of Elaeagnus pungens EmSAMS1 protein from Elaeagnus pungens and a coding gene of the Elaeagnus pungens EmSAMS1 protein in improvement of plant salt tolerance, and belongs to the technical field of gene engineering. According to the invention, elaeagnus pungens is used as a material, and the salt stress-induced EmSAMS1 gene is obtained through cloning. Meanwhile, an overexpression vector is constructed to transform arabidopsis thaliana, which proves that the salt stress resistance of transgenic arabidopsis thaliana is enhanced, and the gene has important application value in the aspects of plant salt-tolerant molecular breeding and variety improvement.

Description

TECHNICAL FIELD

[0001] The present application relates to the field of genetic engineering, and particularly relates to application of EmSAMS1 protein and its coding gene from Elaeagnus oldhamii in improving plant salt tolerance. BACKGROUND

[0002] The saline-alkali soil has poor soil structure, low fertility and is easy to be hardened, thus resulting in decreased soil quality, reduced biodiversity and difficulty for plants to grow normally, and further causing regional ecological environment degradation. The saline-alkali soil can cause osmotic stress, ion toxicity damage and active oxygen damage to plants, and is one of the main non-biological stress factors affecting plant growth, development and reproduction. Mining salt-tolerant genes of plants, analyzing salt-tolerance mechanism of salt-tolerant plants, and breeding and popularizing the use of salt-tolerant plants have become the main way for development and utilization of saline-alkali land.

[0003] Elaeagnus oldhamii Elaeagnus macrophylla Thunb. is an evergreen straight shrub of Elaeagnaceae and Elaeagnus, and is a precious ornamental plant. It is distributed in China, Japan and Korea. In China, it is distributed in coastal islands of Shandong, Jiangsu and Zhejiang and Taiwan. E. oldhamii has strong resistance to sea wind and sea fog, and has developed root system, and is a good tree species for construction of coastal protection forest and greening of low mountain bare rock. In the coastal area of Shandong, E. oldhamii can be used as an optimal species for combination of saline-alkali land restoration and characteristic planting, and has ecological benefits and economic potential. Research on salt-tolerance genes of E. oldhamii and analysis of salt-tolerance mechanism thereof have important roles for development and utilization of plants in saline-alkali land.

[0004] SAMS1 S-adenosylmethionine synthetase (SAMS) is a key enzyme for catalyzing reaction of methionine and ATP to synthesize S-adenosylmethionine (SAM), and plays an important role in regulating dynamic balance of SAM in animals and plants. In plants, SAM as an active primary metabolite provides a methyl donor for methylation of DNA, RNA, protein, hormone and lipid, and directly participates in biosynthesis of substances such as polyamine, ethylene and nicotinamide, and widely participates in regulation of physiological processes such as seed germination, growth and development, flowering and aging of plants. However, the research on related functional genes in E. oldhamii is still relatively less. SAMS1 SUMMARY

[0005] In view of the above prior art, the present application aims to provide application of EmSAMS1 protein and its coding gene from E. oldhamii in improving plant salt tolerance.

[0006] To achieve the above object, the present application adopts the following technical scheme: In a first aspect of the present application, there is provided EmSAMS1 ​The gene is used in the following (1) or (2): (1) improving the tolerance of plants to salt stress; (2) breeding salt-tolerant plant varieties; The EmSAMS1 The gene is a nucleic acid molecule as shown in (i) or (ii) below: (i) a nucleic acid molecule with a nucleotide sequence as shown in SEQ ID NO. 1; (ii) a nucleic acid molecule other than (i) encoding an amino acid sequence as shown in SEQ ID NO. 2.

[0007] In the above application, the tolerance of plants to salt stress is improved by overexpressing the EmSAMS1 gene to promote antioxidant enzyme activity and the accumulation of osmotic regulators.

[0008] In the above application, the plant is Arabidopsis thaliana or Elaeagnus macrophylla.

[0009] The present application first found that the EmSAMS1 gene has a positive regulatory effect on the ability of plants to tolerate salt stress, and under salt stress conditions, the germination rate and main root length of plants with high expression of the EmSAMS1 gene are better than those of wild type. EmSAMS1 It can be used as a new salt-tolerant gene to provide important genetic resources for improving the salt tolerance of plants.

[0010] In a second aspect of the present application, the application of EmSAMS1 protein in improving the salt tolerance of plants is provided; the EmSAMS1 protein is a protein as shown in (A1) or (A2) below: (A1) a protein with an amino acid sequence as shown in SEQ ID NO. 2; (A2) a protein obtained by connecting a protein tag to the N-terminus and / or C-terminus of the protein defined in (A1).

[0011] In the above protein, the protein tag refers to a polypeptide or protein fused and expressed with the target protein by DNA in vitro recombination technology to facilitate the expression, detection, tracking and / or purification of the target protein. The protein tag can be a Flag tag, a His tag, an MBP tag, an HA tag, a myc tag, a GST tag and / or a SUMO tag, etc.

[0012] In the above application, the improvement of the salt tolerance of plants specifically refers to improving the seed germination rate and / or main root length of plants under salt stress treatment.

[0013] In a third aspect of the present application, a method for improving the salt tolerance of plants is provided. EmSAMS1A substance that promotes expression of the gene or a substance that increases the activity and / or content of the EmSAMS1 protein is used in (1) or (2) below: (1) to increase the salt stress tolerance of a plant; (2) to breed a salt-tolerant plant variety.

[0014] In some preferred embodiments of the present application, the substance that promotes expression of the gene is EmSAMS1 The substance that promotes expression of the gene can be any of the following: C1) an expression cassette containing the gene; EmSAMS1 C2) a recombinant vector containing the gene, or a recombinant vector containing the expression cassette of C1); C3) a recombinant microorganism containing the gene, or a recombinant microorganism containing the expression cassette of C1), or a recombinant microorganism containing the recombinant vector of C2); EmSAMS1 C4) a transgenic plant cell line containing the gene, or a transgenic plant cell line containing the expression cassette of C1). EmSAMS1 In a fourth aspect of the present application, a method for increasing the salt stress tolerance of a plant is provided, comprising the step of overexpressing the gene in the plant. EmSAMS1 In the above method, the overexpression of the gene in the plant is achieved by the following means: introducing a gene from an external source; or introducing a DNA fragment capable of activating or increasing the transcription level or translation level or protein activity of the gene.

[0015] EmSAMS1 Advantages of the present application: The present application uses Elaeagnus macrophylla as a material to clone a gene that is induced by salt stress. Meanwhile, an overexpression vector is constructed to transform Arabidopsis thaliana, which proves to enhance the salt stress resistance of the transgenic Arabidopsis thaliana. The present application has important application value in plant salt-tolerant molecular breeding and variety improvement.

[0016] BRIEF DESCRIPTION OF DRAWINGS EmSAMS1 EmSAMS1 : Cloning electropherogram of Elaeagnus macrophylla gene; in the figure, lanes 1-4 are PCR reaction products of Elaeagnus macrophylla gene. EmSAMS1

[0017] EmSAMS1 Advantages of the present application: BRIEF DESCRIPTION OF DRAWINGS

[0018] Figure 1 : Cloning electropherogram of Elaeagnus macrophylla gene; in the figure, lanes 1-4 are PCR reaction products of Elaeagnus macrophylla gene. EmSAMS1 EmSAMS1

[0019] Figure 2 ​​​​​​​​: The pRI101 vector enzyme digestion identification figure; in the figure, lane 1 is the PCR reaction product of the pRI101 vector obtained by double enzyme digestion; lanes 2-4 are the PCR reaction products of the pRI101 vector without double enzyme digestion.

[0020] Figure 3 : The PCR test result of the pRI101-EmSAMS1 recombinant vector monoclonal E. coli.

[0021] Figure 4 : Overexpression EmSAMS1 The PCR and qPCR identification figure of the Arabidopsis thaliana plant.

[0022] Figure 5 : The germination situation figure of the different strains in example 4 of the application under salt stress.

[0023] Figure 6 : The main root length growth situation figure of the different strains in example 4 of the application under salt stress Figure 7 : The phenotype change figure of the different strains in example 4 of the application under salt stress.

[0024] Figure 8 : The physiological and biochemical index determination result figure of the different strains in example 4 of the application under salt stress. DETAILED DESCRIPTION

[0025] It should be noted that the following detailed description is exemplary and is intended to provide further explanation of the present application. Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this application belongs.

[0026] As described previously, researching the salt-tolerant gene in Elaeagnus macrophylla and analyzing the salt-tolerance mechanism have important effects on the development and utilization of plants in saline-alkali land.

[0027] The present application takes Elaeagnus macrophylla as the material, and clones a salt stress-induced gene EmSAMS1 The nucleotide sequence of the gene is shown as SEQ ID NO. 1. EmSAMS1 The amino acid sequence of the EmSAMS1 protein encoded by the gene is shown as SEQ ID NO. 2; and the specific sequence is as follows: EmSAMS1 The gene:

[0028] EmSAMS1 protein: METFLFTSESVNEGHPDKLCDQVSDAVLDACLAQDPDSKVACETCTKTNMVMVFGEITTKATIDYEKIVRDTCRNIGFVSDDVGLDADNCKVLVNIEQ QSPDIAQGVHGHLSKRPEEIGAGDQGHMFGYATDETPELMPLSHVLATKIGAKLTEVRKNGTCPWLRPDGKTQVTVEYYNDKGAMVPIRVHTVLISTQ HDETVTNDEIATDLKKHVIKPVVPEKYLDEKTIFHLNPSGRFVIGGPHGDAGLLTGRKIIIDTYGGWGAHGGGAFSGKDPTKVDRSGAYIVRQAAKSIVASGLAKRCIVQVSYAIGVPEPLSVFVDTYGTGSIPDKEILKIVKESFDFRPGMIAINLDLKRGGNGRRFLKTAAYGHFGRDDSDFTWEVVKPLKWDKVQA .

[0029] To verify EmSAMS1 The function of genes, this invention constructs EmSAMS1 Gene overexpression vectors were developed, and overexpression vectors were created using Agrobacterium-mediated transformation. EmSAMS1 Transgenic Arabidopsis thaliana was used to synthesize genes; then, the germination rate and taproot length of the transgenic Arabidopsis thaliana under salt stress were examined. The results showed that the germination rate and taproot length of the transgenic Arabidopsis thaliana under salt stress were both superior to those of the wild type; further research revealed that: EmSAMS1 Arabidopsis thaliana improves its tolerance to salt stress by alleviating cell membrane damage, promoting antioxidant enzyme activity, and accumulating osmotic regulators.

[0030] To enable those skilled in the art to better understand the technical solution of this application, the technical solution of this application will be described in detail below with reference to specific embodiments.

[0031] In this embodiment of the invention, the experimental materials required for salt tolerance phenotype studies (such as conventional biochemical reagents and plant culture consumables) are all conventional materials for plant stress tolerance research and can be legally obtained through commercial channels. Specific experimental conditions and operating procedures are generally performed according to conventional conditions for plant salt tolerance research, and it is recommended that the experimental reagents and instruments used be performed according to the operating instructions provided by the manufacturer.

[0032] Example 1: Elaeagnus macrocarpa EmSAMS1 Gene cloning and identification 1. Extraction of total RNA from Elaeagnus oldhami leaves: The centrifuge tubes, gun heads and other plastic consumables used in the RNA extraction experiment were all enzyme-free. The mortar and pestle were sterilized in a 180°C oven for 8 hours. The total RNA was extracted from the leaves of Elaeagnus oldhami according to the method of the FastPure Universal Plant Total RNA Isolation Kit of Novagen.

[0033] 2. Reverse transcription to synthesize cDNA: The total RNA extracted from the leaves of Elaeagnus oldhami was reverse transcribed to synthesize cDNA according to the method described in the TVazyme Reverse Transcription Kit.

[0034] 3、 EmSAMS1 Gene cloning: The cDNA synthesized by reverse transcription was used as the template, and specific primers EmSAMS1-F and EmSAMS1-R with SalI and Smal enzyme digestion sites were designed for PCR reaction. The sequences of the specific primers are as follows: EmSAMS1-F: TTGATACATATGCCCGTCGACATGGAGACCTTTTTGTTCACCTCC; EmSAMS1-R: TTCGGATCCGGTACCCCCGGGCTAAGCTTGAACCTTGTCCCACTT.

[0035] The PCR reaction system is shown in Table 1.

[0036] Table 1: PCR reaction system The PCR reaction program is shown in Table 2.

[0037] Table 2: PCR reaction program 4、 EmSAMS1 Gene gel recovery: The PCR reaction product obtained by cloning was subjected to agarose (1%) gel electrophoresis, and then recovered according to the method of the DNA Gel Recovery Kit of Tiangen. The sequence of the PCR reaction product was sequenced and verified.

[0038] The agarose (1%) gel electrophoresis of the PCR reaction product is shown in Figure 1 The PCR amplification product was a fragment of about 1182 bp. The sequencing results of the PCR reaction product showed that the nucleotide sequence was as shown in SEQ ID NO. 1.

[0039] Example 2: Construction of pRI101-EmSAMS1 recombinant expression vector The plasmid DNA of plant eukaryotic expression vector pRI101 was double digested with Sal I and Smal, and the digested plasmid was subjected to PCR detection, and the results are shown in Figure 2 .

[0040] The digested vector pRI101 and the recovered EmSAMS1 gene containing Sal I and Smal homologous arms in Example 1 were ligated, and the ligation reaction system is shown in Table 3. EmSAMS1

[0041] Table 3: Ligation reaction system The above mixture was placed in a 37°C constant temperature water bath for 30 min, and then cooled at 4°C or immediately placed on ice. The ligation product was prepared.

[0042] The ligation product was transformed into E. coli competent DH5α, and then placed in an ice water bath for 30 min, followed by 42°C water bath heat shock for 45 sec. Immediately after heat shock, the ligation product was transferred into an ice water bath for 2-3 min. 900 μL of LB liquid medium without antibiotics was added to the competent cells, and the cap was sealed with sealing film. The mixture was cultured at 37°C and 200 rpm for 1 h. The bacteria were collected by centrifugation at 5000 rpm for 5 min, resuspended with a small amount of supernatant, and evenly coated on LB solid medium containing kanamycin. The medium was incubated in a 37°C incubator for 12-16 h. After the colonies grew, single colonies were selected, and positive strains were screened by colony PCR and sequencing.

[0043] The results are shown in Figure 3 . Clones 1-4 showed a band of about 1182 bp, indicating that the pRI101-EmSAMS1 recombinant expression vector was successfully constructed.

[0044] Example 3: Creation of Arabidopsis thaliana overexpressing EmSAMS1 gene EmSAMS1 1. Preparation of recombinant Agrobacterium infection solution: ​​The pRI101-EmSAMS1 recombinant expression vector constructed in Example 2 was transformed into Agrobacterium competent GV3101, the competent was placed in ice water bath for 45 min, then immediately placed in liquid nitrogen for 1 min; immediately after taking out from liquid nitrogen, heat shock in 37°C water bath for 3 min, then immediately ice water bath for 3 min; 900 μL YEP liquid medium without antibiotics was added to the competent, 28°C shaker culture for 3 h, 10000 rpm centrifugal for 1 min, the bacteria were collected, the bacteria were resuspended with a small amount of supernatant, the bacteria liquid was evenly coated on YEP solid medium containing kanamycin (50 μg / mL) and rifampicin (100 μg / mL) resistance, and cultured in 28°C incubator for 2 days. After the colonies grew, single colony bacteria were selected, and positive strains were screened by colony PCR.

[0045] The obtained positive Agrobacterium strain was inoculated in YEP liquid medium containing kanamycin (50 μg / mL), 28°C, 200 rpm shaking culture for 8 h, until the OD 600 value was 0.8.

[0046] The activated bacteria liquid was divided into 50 mL centrifuge tubes, 4°C, 6000 rpm, centrifugal for 5 min, the bacteria were resuspended with the prepared infection liquid, and the infection liquid preparation system is shown in Table 4.

[0047] Table 4: Infection liquid configuration system 2, Agrobacterium-mediated Arabidopsis genetic transformation (inflorescence infection method): (1) Wild-type Arabidopsis was cultured until the main inflorescence grew, the top main inflorescence was cut off to promote the generation of lateral inflorescence, and the inflorescence was ready for infection and transformation when all the inflorescences were in the white stage.

[0048] (2) The Arabidopsis bracts in the white state were completely immersed in the prepared recombinant Agrobacterium infection liquid, and taken out after 15 s, and the infection was completed.

[0049] (3) The infected bract part was moisturized with plastic wrap, cultured in the dark, and the film was removed after 24 h and placed in a light incubator for normal culture.

[0050] (4) After 7 days, repeat the above steps for infection again; after one month, collect the seeds.

[0051] 3, Transgenic Arabidopsis seed screening: (1) Select an appropriate amount of transgenic Arabidopsis seeds and place them in a 1.5 mL centrifuge tube; add 1 mL of 75% anhydrous ethanol to the centrifuge tube, shake the centrifuge tube vigorously, sterilize for 8 min, 12000 rpm, micro centrifugal for 10 sec, discard the supernatant. Wash the seeds with sterile water 4-5 times.

[0052] (2) Spread the seeds evenly on MS solid medium (containing 50 μg / mL kanamycin), vernalize them in a refrigerator at 4°C for 2 days, and then place them in a light culture room (22°C, 16h light / 8h dark) for 10 days.

[0053] (3) Transplant the resistant seedlings into vermiculite to continue growing. After about 2 months, harvest the T1 generation seeds from the T0 plants.

[0054] 4. Overexpression EmSAMS1 Obtaining and identifying transgenic Arabidopsis thaliana plants: The collected T1 generation seeds were inoculated on MS solid medium, vernalized at 4°C for 2 days, and then transplanted into vermiculite to continue growth, thus obtaining transgenic lines 0E1-0E6.

[0055] When the plant had grown six true leaves, genomic DNA was extracted from the leaves using the Vazyme kit. PCR amplification was performed using specific primers EmSAMS1-F and EmSAMS1-R. The amplified products were analyzed by gel electrophoresis, and the results are as follows: Figure 4 As shown in Figure A, the results indicate that all transgenic lines 0E1-0E6 exhibit a 1182bp fragment; the WT and empty vector lines show no bands.

[0056] Total RNA was extracted from transgenic lines 0E1-0E6, empty vector, and wild-type Arabidopsis thaliana. After purification, cDNA was obtained by reverse transcription using HiScriptⅢ qRT SuperMix for qPCR (+gDNA wiper) reverse transcriptase.

[0057] Arabidopsis thaliana AtActin The gene is an internal reference gene. Primers for the internal reference gene were designed, and their sequences are as follows: AtActin- F: GATGCCCAGAAGTCTTGTTCCAG; AtActin- R: CTTTGCTCATACGGGTCAGCGATAC.

[0058] Designed using Primer 5.0 EmSAMS1 The specific primers for quantitative real-time PCR, qPCR-EmSAMS1-F and qPCR-EmSAMS1-R, have the following primer sequences: qPCR-EmSAMS1-F:ATGGAGACCTTTTTGTTCACCTCCG; qPCR-EmSAMS1-R:CGGAGGTGAACAAAAAGGTCTCCAT.

[0059] Using qRT-PCR primers for different strains EmSAMS1 Genes were detected using real-time quantitative PCR. Results analysis employed a comparative threshold method to quantify the real-time quantitative PCR results. A fluorescence threshold was set, and the cycle number Ct was determined within that threshold. The C value was calculated based on the Ct value. C=2 -△△Ct , △Ct=Ct 目的基因 -Ct 内参基因 The average C value of the three replicates was calculated as the relative expression level of the target gene.

[0060] The results of qRT-PCR detection are as follows Figure 4 As shown in B, the results indicate that in transgenic lines 0E1-0E6... EmSAMS1 Gene expression levels were all significantly upregulated.

[0061] In summary, this invention successfully constructs an overexpression... EmSAMS1 Transgenic Arabidopsis thaliana plants with the gene. OE1 was selected as the material for subsequent functional validation studies.

[0062] Example 4: Overexpression EmSAMS1 Salt tolerance study of transgenic Arabidopsis thaliana 1. Test method: (1) Germination rate assessment: OE1 homozygous T3 generation transgenic Arabidopsis seeds were selected for experiments. Wild-type Arabidopsis (WT), empty vector (pRI101), and overexpression were used to express the transgenic Arabidopsis seeds. EmSAMS1 The seeds of Arabidopsis thaliana were sown on MS medium and 100 mM NaCl + MS medium. Germination was observed after 5 days and the germination rate was recorded.

[0063] (2) Examination of taproot length: To analyze root development after germination, wild-type Arabidopsis thaliana (WT), the empty vector (pRI101), and the overexpressing rheumatoid arthritis were first introduced into the root system. EmSAMS1 After vernalization at 4°C for 3 days, Arabidopsis thaliana seeds were sown on MS medium. After growing for 7 days under normal light conditions, the seedlings were transferred to MS and 100mM NaCl+MS medium. Root growth was observed on the 10th and 20th days, and the length of the taproot was measured.

[0064] (3) Investigation of plant phenotype and physiological indicators under salt treatment conditions: Wild-type Arabidopsis thaliana (WT), empty vector (pRI101), and overexpression were respectively used. EmSAMS1After vernalization at 4°C for 3 days, Arabidopsis thaliana seeds were sown on MS medium and cultured in a light incubator for 12 days. Seedlings with uniform growth were selected, transplanted into vermiculite, and cultured in an incubator (16 h day / 8 h night, 22°C, 60% relative humidity) for another 3 weeks. Then, a 200 mM NaCl solution was prepared for salt stress treatment. The seedlings were watered with this salt solution daily to ensure that there was always salt solution seeping out of the tray. Samples were taken at 0, 6, 12, 24, and 72 h after salt stress, flash-frozen in liquid nitrogen, and stored at -80°C for subsequent physiological experiments (determination of MDA, SOD, POD, and proline). Phenotypic changes in Arabidopsis thaliana were also photographed and recorded.

[0065] 2. Test Results: On NaCl-free MS medium, germination rate and taproot length were observed in wild-type, empty vector, and overexpression samples. EmSAMS1 There were no significant differences among the plants. However, on MS medium containing 100 mM NaCl, overexpression... EmSAMS1 Arabidopsis thaliana plants had better germination rates and taproot lengths than wild-type and empty-carrier plants. Figure 5 and Figure 6 ).

[0066] Treatment with 200 mM NaCl significantly stressed wild-type and empty vector-transformed Arabidopsis plants: leaf color deepened, and some leaves yellowed or even withered. Overexpression... EmSAMS1 Arabidopsis showed almost no significant changes. Figure 7 ).

[0067] Malondialdehyde (MDA) is a key product of membrane lipid peroxidation and can be used to measure the degree and extent of membrane lipid peroxidation and damage under stress. Antioxidant enzymes such as superoxide dismutase (SOD) and peroxidase (POD) are major components of the enzymatic antioxidant system that scavenges reactive oxygen species (ROS), playing a crucial role in clearing excess ROS generated under stress and regulating ROS balance. Proline enhances salt tolerance through osmotic regulation, antioxidant activity, and ion homeostasis. In this study, the contents of MDA, SOD, POD, and proline in different strains changed significantly at 0, 6, 12, 24, and 72 hours after salt stress. The MDA content in each strain showed a trend of first increasing and then decreasing after salt stress, and overexpression of proline increased significantly within 72 hours of salt stress. EmSAMS1 The MDA content in the strains was consistently lower than that in the wild-type and empty vector strains. Figure 8 -A). Comparing the differences in antioxidant enzyme activities, it was found that both SOD and POD activities showed a trend of first increasing and then decreasing, and EmSAMS1 The enzyme activities of the transgenic lines were significantly higher than those of the control at all time points. Figure 8B, C). The SOD and POD activities in the transgenic lines reached the maximum at 24 h of salt stress. Under salt stress, the proline content in each line gradually increased within 72 h and reached the maximum at 72 h. The proline content in the transgenic lines reached 181.37% of that in the WT under the same conditions Figure 8 D). This indicates EmSAMS1 By alleviating the degree of cell membrane damage, promoting antioxidant enzyme activity and the accumulation of osmotic adjustment substances, the salt tolerance of Arabidopsis was improved.

[0068] In summary, the gene of Elaeagnus macrophylla Turcz. EmSAMS1 EmSAMS1 EmSAMS1 EmSAMS1 EmSAMS1 EmSAMS1 EmSAMS1 EmSAMS1 EmSAMS1 EmSAMS1 EmSAMS1 EmSAMS1 EmSAMS1 EmSAMS1 EmSAMS1 EmSAMS1 EmSAMS1 EmSAMS1 EmSAMS1 EmSAMS1 EmSAMS1 EmSAMS1 EmSAMS1 EmSAMS1 EmSAMS1 EmSAMS1 EmSAMS1 EmSAMS1 EmSAMS1 EmSAMS1 EmSAMS1 EmSAMS1 EmSAMS1 EmSAMS1 EmSAMS1 EmSAMS1 EmSAMS1 EmSAMS1 EmSAMS1 EmSAMS1 EmSAMS1 EmSAMS1 EmSAMS1 EmSAMS1 EmSAMS1 EmSAMS1 EmSAMS1 EmSAMS1 EmSAMS1 EmSAMS1 EmSAMS1 EmSAMS1 EmSAMS1 EmSAMS1 EmSAMS1 EmSAMS1 EmSAMS1 EmSAMS1 EmSAMS1 EmSAMS1 EmSAMS1 EmSAMS1 EmSAMS1 Transformed into the model plant Arabidopsis thaliana, the salt tolerance of transgenic Arabidopsis thaliana can be significantly improved.

[0069] The above only describes the preferred embodiments of the present application and is not intended to limit the present application. For those skilled in the art, the present application can have various modifications and changes. Any modification, equivalent replacement, improvement, etc. made within the spirit and principles of the present application shall be included in the protection scope of the present application.

Claims

1. EmSAMS1 The application of genes in the following (1) or (2): (1) Improve the plant's tolerance to salt stress; (2) Cultivate salt-tolerant plant varieties; The EmSAMS1 A gene is a nucleic acid molecule as shown in (i) or (ii): (i) Nucleic acid molecules with nucleotide sequences as shown in SEQ ID NO.1; (ii) Nucleic acid molecules other than (i) that encode the amino acid sequence shown in SEQ ID NO.

2.

2. The application according to claim 1, characterized in that, Through overexpression EmSAMS1 Genes that enhance plant tolerance to salt stress by promoting antioxidant enzyme activity and the accumulation of osmotic regulators.

3. The application according to claim 1, characterized in that, The plant in question is either Arabidopsis thaliana or Elaeagnus macrophylla.

4. Application of EmSAMS1 protein in improving plant salt tolerance; the EmSAMS1 protein is the protein shown in (A1) or (A2) below: (A1) A protein with the amino acid sequence shown in SEQ ID NO.2; (A2) The protein obtained by attaching a protein tag to the N-terminus and / or C-terminus of the protein defined in (A1).

5. The application according to claim 4, characterized in that, Specifically, improving plant salt tolerance means increasing the seed germination rate and / or taproot length of plants under salt stress treatment.

6. Promote EmSAMS1 The application of substances that enhance gene expression or increase the activity and / or content of EmSAMS1 protein in the following (1) or (2): (1) Improve the plant's tolerance to salt stress; (2) Cultivate salt-tolerant plant varieties.

7. The application according to claim 6, characterized in that, Promote EmSAMS1 The substance expressed by the gene is any one of the following: C1) contains EmSAMS1 Gene expression cassettes; C2) contains EmSAMS1 Recombinant vectors of genes, or recombinant vectors containing the expression cassette described in C1); C3) contains EmSAMS1 Recombinant microorganisms containing genes, or recombinant microorganisms containing the expression cassette described in C1), or recombinant microorganisms containing the recombinant vector described in C2); C4) contains EmSAMS1 Transgenic plant cell lines containing the gene, or transgenic plant cell lines containing the expression cassette described in C1).

8. A method for improving plant tolerance to salt stress, characterized in that, include: In plants EmSAMS1 The steps of gene overexpression.

9. The method according to claim 8, characterized in that, In plants EmSAMS1 Gene overexpression is achieved through the following methods: Exogenous transfer EmSAMS1 Genes; or the introduction of genes that can activate or enhance [something]. EmSAMS1 DNA fragments at the transcriptional or translational level or for protein activity.

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