CN114525298A - Application of soybean protein GmFVE in plant salt tolerance regulation - Google Patents

Abstract

The invention discloses application of soybean protein GmFVE in plant salt tolerance regulation. The invention provides an application of a GmFVE protein in regulating and controlling plant salt tolerance. Specifically, the regulation is negative regulation. Namely, the content and/or activity of the GmFVE protein is reduced, and the salt tolerance of the plant is increased. The invention also provides a method for preparing a transgenic plant with improved salt stress tolerance, which comprises the following steps: a substance which inhibits the expression of a nucleic acid molecule encoding a GmFVE protein is introduced into a receptor plant, so that a transgenic plant with improved salt stress tolerance is obtained. The invention has important theoretical and practical significance for cultivating high-salt-tolerance plant varieties.

Description

Application of soybean protein GmFVE in regulation of plant salt tolerance

technical field

The invention relates to the field of biotechnology, in particular to the application of soybean protein GmFVE in the regulation of plant salt tolerance.

Background technique

Changes in physical and chemical factors in the environment, such as drought, salinity, chilling damage, freezing damage, waterlogging and other stress factors are one of the reasons for serious crop yield reduction. During the 40 years from 1939 to 1978 in the United States, the insurance industry's compensation statistics for crop yield reduction showed that the compensation ratio for yield reduction due to salt damage and drought accounted for about 40.8%, which was higher than that of waterlogging (16.4%) and low temperature (13.8%). , hail (11.3%) and wind (7.0%), which are much higher than insect pests (4.5%), diseases (2.7%) and other factors. Therefore, the cultivation of salt/drought tolerant crops is one of the main goals of the planting industry. To improve the salt/drought tolerance of crops, in addition to using traditional breeding methods, molecular genetic breeding has become one of the fields of concern for scientific and technological workers.

FVE/MSI4 has a WD40 repeat domain, which is highly homologous to the RbAp protein (retinoblastoma-associated protein) in mammals. It is one of the components of the histone deacetylase (HDAC) complex and is involved in transcriptional repression. In Arabidopsis thaliana, FVE/MSI4 regulates plant flowering by inhibiting the expression of FLC by maintaining a low level of histone H3ac in the key flowering gene FLC. There are relatively few functional studies of this protein in soybean. So far, there are no reports on its association with salt tolerance.

SUMMARY OF THE INVENTION

The purpose of the present invention is to provide the application of soybean protein GmFVE in the regulation of plant salt tolerance.

The present invention provides the application of GmFVE protein in regulating the salt tolerance of plants. Specifically, the regulation is negative regulation. That is, reducing the content and/or activity of GmFVE protein increases the salt tolerance of plants. That is, when the content and/or activity of the GmFVE protein is increased, the salt tolerance of the plant is decreased.

The invention also protects the application of biological materials related to GmFVE protein in regulating the salt tolerance of plants;

The biological material related to GmFVE protein is any one of the following (d1) to (d12):

(d1) a nucleic acid molecule encoding a GmFVE protein;

(d2) an expression cassette containing the nucleic acid molecule of (d1);

(d3) a recombinant vector containing the nucleic acid molecule of (d1);

(d4) a recombinant vector containing the expression cassette of (d2);

(d5) a recombinant microorganism containing the nucleic acid molecule of (d1);

(d6) a recombinant microorganism containing the expression cassette of (d2);

(d7) a recombinant microorganism containing the recombinant vector of (d3);

(d8) a recombinant microorganism containing the recombinant vector of (d4);

(d9) a transgenic plant cell line containing the nucleic acid molecule of (d1);

(d10) a transgenic plant cell line containing the expression cassette of (d2);

(d11) a transgenic plant cell line containing the recombinant vector of (d3);

(d12) A transgenic plant cell line containing the recombinant vector of (d4).

Specifically, the regulation is negative regulation. That is, by suppressing the expression of the nucleic acid molecule encoding the GmFVE protein, the salt tolerance of the plant is increased. That is, the expression of the nucleic acid molecule encoding the GmFVE protein is promoted, and the salt tolerance of the plant is reduced.

The present invention also protects the use of substances inhibiting GmFVE protein in plant breeding for the purpose of cultivating plants with increased stress tolerance to salt stress.

The present invention also protects the use of a substance that inhibits the expression of a nucleic acid molecule encoding a GmFVE protein for cultivating transgenic plants with increased stress tolerance to salt stress. The substance that inhibits the expression of the nucleic acid molecule encoding the GmFVE protein can be a substance that inhibits the expression of the nucleic acid molecule encoding the GmFVE protein based on RNAi technology. The substance that inhibits the expression of the nucleic acid molecule encoding the GmFVE protein can be a specific DNA molecule or a recombinant plasmid having the specific DNA molecule. Among the specific DNA molecules, there are DNA molecule A and DNA molecule B. DNA molecule A is shown in Sequence 3 of the sequence table, and DNA molecule B is reverse complementary to DNA molecule A. Specifically, the recombinant plasmid can be as follows: DNA molecule A is inserted between the Sac I and Kpn I restriction sites of the pZH01 vector, and DNA molecule B is inserted between the Sal I and Xba I restriction sites. The specific method of the application is as follows: the substance that inhibits the expression of the nucleic acid molecule encoding the GmFVE protein is introduced into the target plant through Agrobacterium rhizogenes to obtain a chimeric plant with hairy roots; the chimeric plant is resistant to salt stress. Sexual improvement.

The present invention also protects a method for preparing a transgenic plant with improved stress tolerance to salt stress, comprising the steps of: introducing a substance that inhibits the expression of a nucleic acid molecule encoding a GmFVE protein into the recipient plant to obtain stress tolerance to salt stress Enhanced transgenic plants. The substance that inhibits the expression of the nucleic acid molecule encoding the GmFVE protein can be a substance that inhibits the expression of the nucleic acid molecule encoding the GmFVE protein based on RNAi technology. The substance that inhibits the expression of the nucleic acid molecule encoding the GmFVE protein can be a specific DNA molecule or a recombinant plasmid having the specific DNA molecule. Among the specific DNA molecules, there are DNA molecule A and DNA molecule B. DNA molecule A is shown in Sequence 3 of the sequence table, and DNA molecule B is reverse complementary to DNA molecule A. The recombinant plasmid can be specifically as follows: DNA molecule A is inserted between the Sac I and Kpn I restriction sites of the pZH01 vector, and DNA molecule B is inserted between the Sal I and Xba I restriction sites. In the method, a substance that inhibits the expression of a nucleic acid molecule encoding a GmFVE protein is introduced into a recipient plant through Agrobacterium rhizogenes, to obtain a chimeric plant with hairy roots with improved stress tolerance to salt stress.

The present invention also protects a plant breeding method, comprising the steps of: reducing the content and/or activity of GmFVE protein in the target plant, thereby improving the stress tolerance of the target plant to salt stress.

The increased stress tolerance to salt stress is reflected as follows (f1) or (f2):

(f1) Increased survival in salt-stressed environments;

(f2) Membrane damage is alleviated in a salt-stressed environment.

In any of the above recombinant vectors, any enhanced, constitutive, tissue-specific or inducible promoter, such as cauliflower mosaic virus (CAMV) 35S promoter, can be added before the transcription initiation nucleotide. , ubiquitin gene Ubiquitin promoter (pUbi), etc., they can be used alone or in combination with other plant promoters; in any of the above recombinant vectors, enhancers can also be used, including translation enhancers or transcription enhancers , These enhancer regions can be ATG start codons or adjacent region start codons, etc., but must be the same as the reading frame of the coding sequence to ensure the 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 vectors used can be processed, such as adding genes (GUS gene, luciferase gene, etc. ), antibiotic markers with resistance (gentamicin marker, kanamycin marker, etc.) or marker genes for chemical resistance (such as herbicide resistance genes). Considering the safety of transgenic plants, the transformed plants can be directly screened under stress without adding any selectable marker gene.

In any of the above recombinant plasmids, any enhanced, constitutive, tissue-specific or inducible promoter, such as cauliflower mosaic virus (CAMV) 35S promoter, can be added before the transcription initiation nucleotide. , ubiquitin gene Ubiquitin promoter (pUbi), etc., which can be used alone or in combination with other plant promoters; in any of the above recombinant plasmids, enhancers can also be used, including translation enhancers or transcription enhancers . 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 vectors used can be processed, such as adding genes (GUS gene, luciferase gene, etc. ), antibiotic markers with resistance (gentamicin marker, kanamycin marker, etc.) or marker genes for chemical resistance (such as herbicide resistance genes). Considering the safety of transgenic plants, the transformed plants can be directly screened under stress without adding any selectable marker gene.

The recombinant vector or the recombinant plasmid can transform plant cells or tissues by using conventional biological methods such as Ti plasmid, Ri plasmid, plant virus vector, direct DNA transformation, microinjection, electrical conductivity, Agrobacterium-mediated, etc. Plant cells or tissues are grown into plants.

Any of the above-mentioned Agrobacterium rhizogenes may specifically be Agrobacterium rhizogenes K599.

The GmFVE protein described above, obtained from Glycine max (L.) Merrill, is as follows (a) or (b) or (c):

(a) the protein shown in Sequence 1 of the Sequence Listing;

(b) a protein derived from soybean and having more than 98% identity and the same function as the protein shown in SEQ ID NO: 1 of the Sequence Listing;

(c) A protein with the same function obtained by substituting and/or deleting and/or adding one or several amino acid residues to the protein shown in SEQ ID NO: 1 of the sequence listing.

Any one of the above-mentioned nucleic acid molecules encoding GmFVE proteins is as follows (e1), (e2) or (e3):

(e1) a DNA molecule whose coding region is shown in Sequence 2 of the Sequence Listing;

(e2) a DNA molecule having 75% or more identity with (e1) and encoding the GmFVE protein;

(e3) a DNA molecule that hybridizes to (el) or (e2) under stringent conditions and encodes the GmFVE protein.

Any of the above-mentioned plants are monocotyledonous or dicotyledonous. The monocotyledonous plant can be rice, wheat or corn, and the like. The dicotyledonous plant can be soybean, tobacco or cotton and the like. The dicotyledonous plant is specifically soybean, and the soybean is specifically Kefeng No. 1.

In the present invention, GmFVE-RNAi that reduces GmFVE activity is transferred into the hairy roots of recipient soybeans to obtain transgenic hairy roots and chimeric plants. The GmFVE protein was significantly improved, indicating that GmFVE protein negatively regulates plant salt tolerance. The invention has important theoretical and practical significance for cultivating high salt-tolerant plant varieties.

Description of drawings

Figure 1 is a schematic diagram of the structure of the pZH01 vector.

Figure 2 is the molecular identification of GmFVE-RNAi soybean hairy roots.

Figure 3 shows the salt-tolerant phenotype of GmFVE-RNAi soybean hairy roots and chimeras.

Figure 4 shows the relative conductance of leaves of transgenic GmFVE-RNAi hairy root chimeras under high salt stress.

Figure 5 shows the survival rate of transgenic GmFVE-RNAi hairy root chimeras under high salt stress.

Detailed ways

The present invention will be further described in detail below with reference to the specific embodiments, and the given examples are only for illustrating the present invention, rather than for limiting the scope of the present invention. The examples provided below can serve as a guide for those of ordinary skill in the art to make further improvements, and are not intended to limit the present invention in any way.

The experimental methods in the following examples, unless otherwise specified, are conventional methods, and are performed according to the techniques or conditions described in the literature in the field or according to the product specification. The materials, reagents, etc. used in the following examples can be obtained from commercial sources unless otherwise specified.

Unless otherwise specified, the quantitative tests in the following examples are set to repeat the experiments three times, and the results are averaged.

The primers used in the following examples were all synthesized by Sanbo Biological Company.

Glycine max L.Merr.Kefeng 1 in the following examples is described in the following documents: W.K.Zhang, Y.J.Wang, G.Z.Luo, J.S.Zhang, C.Y.He, X.L.Wu, J.Y.Gai, S.Y.Chen , QTLmapping of ten agronomic traits on the soybean(Glycine max L.Merr.) geneticmap and their association with EST markers, Theor.Appl.Genet, 2004, 108:1131-1139; the public can obtain from the Chinese Academy of Sciences Genetics and Developmental Biology Research obtained.

The pZH01 vector, Stratagene Company, is described in the following documents: Han Xiao, et al. Functional analysis of the rice AP3 homologue OsMADS16 by RNA interference, Plant Molecular Biology, 2003, 52, 957-966. A schematic diagram of the pZH01 vector structure is shown in Figure 1.

Agrobacterium rhizogenes K599 is described in the following documents: Attila Kereszt, et al., Agrobacterium rhizogenes-mediaded transformation of soybean to study of root biology, Nature Protocols, 2007, 2(4), 549-552.

Example 1. Discovery of soybean protein GmFVE protein and its encoding gene (GmFVE gene)

The inventor's research found that soybean transcription factor GmNFYA regulates plant salt stress response. GmNFYA was overexpressed in soybean Jack and significantly improved the salt tolerance of transgenic soybean. The inventors used the soybean hairy root system to obtain transgenic hairy roots overexpressing GFP and NFYA-GFP, respectively. The protein interacting specifically with NFYA-GFP was obtained by IP-MS analysis. The related proteins were screened and verified by yeast two-hybrid experiments. It was found that GmNFYA could directly interact with the protein encoded by Glyma.09G063100. BiFC and LUC complementation experiments showed that the two proteins also interacted in vivo. Glyma.09G063100 encodes FVE/MSI4 protein with a WD40 repeat domain, which may be involved in the regulation of soybean salt tolerance.

Total RNA was extracted from soybean Kefeng 1 seedlings, and cDNA was obtained by reverse transcription. Using cDNA as a template, PCR amplification was performed using a primer pair composed of GmFVE-up and GmFVE-dp, and the PCR amplification product was recovered and sequenced, as shown in Sequence 2 of the sequence table. The DNA molecule shown in Sequence 2 of the Sequence Listing encodes the protein shown in Sequence 1 of the Sequence Listing. The protein shown in SEQ ID NO: 1 of the Sequence Listing is named GmFVE protein, and consists of 513 amino acid residues. The gene encoding the GmFVE protein was named GmFVE gene.

GmFVE-up: 5'-ATGGAGACTCCTCCTCCCCAACAAG-3';

GmFVE-dp: 5'-TCATTTTTCAGTCTTTGAAGCACAC-3'.

Example 2. Construction of pZH01-GmFVE-RNAi plant vector

1. Extract the total RNA of soybean Kefeng No. 1 seedlings, and reverse-transcribe to obtain cDNA.

2. Using the cDNA obtained in step 1 as a template, a primer pair composed of FVE-RNAi-F and FVE-RNAi-R is used for PCR amplification, and the PCR amplification product is recovered.

FVE-RNAi-F: 5'-TGC TCTAGA GAGCTC ACGACTGGCTCGCCAACCAC-3';

FVE-RNAi-R: 5'-ACGC GTCGAC GGTACC ACTGTTTTGTCCTTTCCTCCTG-3'.

3. Take the PCR amplification product obtained in step 2, carry out double digestion with restriction enzymes Sac I and Kpn I, and recover the digestion product.

4. Take the pZH01 vector, carry out double digestion with restriction enzymes Sac I and Kpn I, and recover the vector backbone.

5. Connect the enzyme cut product obtained in step 3 and the vector backbone obtained in step 4 to obtain a recombinant plasmid.

6. Take the PCR amplification product obtained in step 2, carry out double digestion with restriction endonucleases Xba I and Sal I, and recover the digestion product.

7. Take the recombinant plasmid obtained in step 5, carry out double digestion with restriction enzymes Sal I and Xba I, and recover the vector backbone.

8. Connect the digested product obtained in step 6 with the vector backbone obtained in step 7 to obtain a recombinant plasmid.

9. Take the recombinant plasmid obtained in step 8 and perform sequencing verification. The sequencing results showed that: in the recombinant plasmid, DNA molecule A was inserted between the Sac I and Kpn I restriction sites of the pZH01 vector, and DNA molecule B was inserted between the Sal I and Xba I restriction sites; DNA molecule As shown in Sequence 3 of the sequence listing, DNA molecule B is reverse complementary to DNA molecule A. The recombinant plasmid was named pZH01-GmFVE-RNAi plant vector.

Example 3, the acquisition of chimeric plants

Agrobacterium rhizogenes infection method is slightly improved according to methods such as Attila Kereszt (Attila Kereszt, et al., Agrobacterium rhizogenes-mediaded transformation of soybean to study of rootbiology, Nature Protocols, 2007, 2 (4), 549-552), For reference, Wang, Fang; Chen, Hao-Wei; Li, Qing-Tian; Wei, Wei; Li, Wei; Zhang, Wan-Ke; Ma, Biao; Bi, Ying-Dong; Lai, Yong-Cai; Liu, Xin-Lei; Man, Wei-Qun; Zhang, Jin-Song; Chen, Shou-Yi, GmWRKY27 interacts with GmMYB174 to reduce expression of GmNAC29 for stress tolerance in soybeanplants, 2015, The Plant Journal, 83, 224–236, or authorized patents, Chen Shouyi et al., Plant stress tolerance-related transcription factor GmWRKY78 and its encoding gene and application, grant number: ZL2011 1 0053083.7, grant date 2013.10.09.

1. Obtainment of recombinant Agrobacterium

The pZH01-GmFVE-RNAi plant vector was introduced into Agrobacterium rhizogenes K599 by electroporation to obtain a recombinant Agrobacterium, which was named as recombinant Agrobacterium K599/pZH01-GmFVE-RNAi. Recombinant Agrobacterium K599/pZH01-GmFVE-RNAi was inoculated into liquid LB medium containing 50 mg/L hygromycin and cultured until the system OD600nm value was above 1.0.

The pZH01 vector was introduced into Agrobacterium rhizogenes K599 by electroporation to obtain a recombinant Agrobacterium, which was named as recombinant Agrobacterium K599/pZH01. Recombinant Agrobacterium K599/pZH01 was inoculated into liquid LB medium containing 50 mg/L hygromycin and cultured until the OD600nm value of the system was above 1.0.

2. Preparation of transgenic chimeric plants

The seeds of soybean Kefeng No. 1 were sown in vermiculite and cultivated until 2 true leaves were formed. Use a syringe to inoculate the recombinant Agrobacterium K599/pZH01-GmFVE-RNAi bacterial solution prepared in step 1 to the base of the plant stem (about 50 microliters per plant), using "light for 16 hours/dark for 8 hours, temperature 25 ℃, humidity 50%” conditions were cultured for 2 weeks, at which time the plants had grown hairy roots, and a total of 100 plants with hairy roots were obtained, which were named as transgenic chimeric plants.

3. Preparation of empty vector chimeric plants

The seeds of soybean Kefeng No. 1 were sown in vermiculite and cultivated until 2 true leaves were formed. Use a syringe to inoculate the recombinant Agrobacterium K599/pZH01 bacterial solution prepared in step 1 to the base of the plant stem (about 50 microliters per plant), using "light for 16 hours/dark for 8 hours, temperature 25 ℃, humidity 50%". Conditional culture was carried out for 2 weeks, at which time the plants had grown hairy roots. A total of 98 plants with hairy roots were obtained, which were named as empty vector chimera plants.

4. The level of GmFVE gene in hairy roots

Test samples: hairy roots of transgenic chimeric plants or hairy roots of chimeric plants transduced with an empty vector.

Total RNA from the test sample was extracted and reverse transcribed into cDNA. Quantitative PCR was performed with primer pairs consisting of FVE-qPCR-F and FVE-qPCR-R with cDNA as template to detect the relative expression level of GmFVE gene. The soybean GmTubulin gene was used as the internal standard, and the primer pair used to detect the internal standard consisted of Primer-TF and Primer-TR.

FVE-qPCR-F: 5'-GGAAGGGTTTGAAGGAAGGTAGG-3';

FVE-qPCR-R: 5'-TCGTAGAGGACAGGAACAAGGGA-3'.

Primer-TF: 5'-AACCTCCTCCTCATCGTACT-3';

Primer-TR: 5'-GACAGCATCAGCCATGTTCA-3'.

The results are shown in Figure 2. The level of GmFVE gene in the hairy roots of the transgenic chimeric plants was taken as 1, and the relative expression level of the GmFVE gene in the hairy roots of the transgenic chimeric plants was only 0.38.

Example 4. Identification of plant salt tolerance

Test plants: The transgenic chimeric plants or the empty vector chimeric plants prepared in Example 3 are all plants 12 days after inoculation with the bacterial solution.

One, group processing

The test plants were divided into 2 groups with 15 plants in each group. The first group (salt stress group): 100 mM NaCl aqueous solution was used to hydroponic test plants (plant roots were immersed in NaCl aqueous solution). The second group (control group): the 100 mM NaCl aqueous solution was replaced with water, and the others were the same as the first group.

Photographs were taken after 3 days of treatment, as shown in Figure 3. Control group: There was no significant difference in phenotype between transgenic chimeric plants and empty vector chimeric plants. Salt stress group: the leaves (especially the lower leaves) of the chimera plants transformed with the empty vector had withered or started to wilt, and the leaves of the transgenic chimera plants were basically not wilted.

2. After 3 days of treatment in step 1, the relative ion permeability was detected.

When plant tissue is damaged by stress, the cell membrane function is damaged or the structure is damaged, and the permeability is increased, so that various water-soluble substances in the cell, including electrolytes, are extravasated. Immersion of plant tissue in deionized water increases the conductance of the water due to the extravasation of electrolytes. The heavier the damage, the greater the damage to the cell membrane, the greater the extravasation, and the greater the conductivity of the water. Therefore, the conductivity meter can be used to measure the change of the conductivity of the extravasation fluid, which indirectly reflects the degree of damage to the plant tissue. Therefore, the detection of electrical conductivity can calculate the relative ion permeability, which indicates the degree of damage to the plant cell membrane.

Detection method: Cut the leaves of the plant, put them in a clean screw-top glass bottle, rinse with deionized water 3 times; then add 80 mL of deionized water to completely soak the leaves, and vacuumize for 45 minutes; then stand at room temperature for 30 minutes; Then, the leaves were boiled for 15 min, after the temperature dropped to room temperature, mixed well, and the conductivity E2 was measured with a conductivity meter.

Relative ion permeability EL(%)=E1/E2×100.

The results are shown in Figure 4. Control group: The relative ion permeability of leaves of transgenic chimera plants and empty vector chimera plants were 8% and 10%, respectively, with no significant difference. Salt stress group: the relative ion permeability of the leaves of the transgenic chimeric plants was 40%, the relative ion permeability of the leaves of the vector chimera plants was 68%, and the damage to the cell membrane of the transgenic chimera plants was significantly less than that of the empty vector chimera plants. Vector chimeric plants.

3. After 7 days of treatment in step 1, the survival rate was counted.

The results of the survival rate of the salt stress group are shown in Figure 5. The survival rate of transgenic chimera plants was 71%, and the survival rate of chimera plants transformed with empty vector was 32%. The survival rate of transgenic chimera plants was significantly higher than that of chimera plants transformed with empty vector.

All the above results indicated that inhibiting the expression of GmFVE gene could significantly improve the salt tolerance of plants.

The present invention has been described in detail above. For those skilled in the art, without departing from the spirit and scope of the present invention, and without unnecessary experimentation, the present invention can be implemented in a wide range under equivalent parameters, concentrations and conditions. Although the present invention has given particular embodiments, it should be understood that the present invention can be further modified. In conclusion, in accordance with the principles of the present invention, this application is intended to cover any alterations, uses or improvements of the present invention, including changes made using conventional techniques known in the art, departing from the scope disclosed in this application. The application of some of the essential features can be made within the scope of the following appended claims.

sequence listing

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acgactggct cgccaaccac aacctcgtct ggccctctct ctcttgcagg tggggccccc 60

agcttgaaca agccacttac aagaatcgcc agagactcta cctttctgag cagactgatg 120

gtagtgtgcc gaatactctg gtgattgcga attgcgaggt tgtgaagcct agggttgctg 180

ctgctgagca catttcgcag tttaatgaag aggcgcggtc cccatttgtg aagaagtaca 240

agaccatcat acatcctggt gaggtaaaca gaattaggga attgccacaa aattccaaga 300

tagtggctac acatacagac agccctgatg tccttgtttg ggatgttgaa agtcaaccta 360

atcgccatgc tgtccttgga gctacaaact ctcgtcctga tttgatattg accggacacc 420

aagataatgc ggaatttgct cttgctatgt gcccaactga accctatgtt ctttcaggag 480

gaaaggacaa aacagt 496

Claims (10)

1. the application of the substance that suppresses GmFVE protein in plant breeding, the goal of described breeding is to cultivate the plant that the stress tolerance to salt stress improves; The GmFVE protein is as follows (a) or (b) or (c): (a) the protein shown in Sequence 1 of the Sequence Listing; (b) a protein derived from soybean and having more than 98% identity and the same function as the protein shown in SEQ ID NO: 1 of the Sequence Listing; (c) A protein with the same function obtained by substituting and/or deleting and/or adding one or several amino acid residues to the protein shown in Sequence 1 of the sequence listing. 2. the application of the substance that suppresses the expression of the nucleic acid molecule encoding the GmFVE protein, the application is to cultivate the transgenic plant with improved stress tolerance to salt stress; The GmFVE protein is as follows (a) or (b) or (c): (a) the protein shown in Sequence 1 of the Sequence Listing; (b) a protein derived from soybean and having more than 98% identity and the same function as the protein shown in SEQ ID NO: 1 of the Sequence Listing; (c) A protein with the same function obtained by substituting and/or deleting and/or adding one or several amino acid residues to the protein shown in Sequence 1 of the sequence listing. 3. a method for preparing a transgenic plant with improved stress tolerance to salt stress, comprising the steps of: importing a material that suppresses the expression of the nucleic acid molecule encoding the GmFVE protein in a recipient plant, to obtain an improved stress tolerance to salt stress. transgenic plants; The GmFVE protein is as follows (a) or (b) or (c): (a) the protein shown in Sequence 1 of the Sequence Listing; (b) a protein derived from soybean and having more than 98% identity and the same function as the protein shown in SEQ ID NO: 1 of the Sequence Listing; (c) A protein with the same function obtained by substituting and/or deleting and/or adding one or several amino acid residues to the protein shown in Sequence 1 of the sequence listing. 4. The application of claim 2 or the method of claim 3, wherein: Nucleic acid molecules encoding GmFVE proteins are as follows (e1), (e2) or (e3): (e1) a DNA molecule whose coding region is shown in Sequence 2 of the Sequence Listing; (e2) a DNA molecule having 75% or more identity with (e1) and encoding the GmFVE protein; (e3) a DNA molecule that hybridizes to (el) or (e2) under stringent conditions and encodes the GmFVE protein. 5. a plant breeding method, comprising the steps: reduce the content and/or activity of GmFVE protein in the target plant, thereby making the target plant improve the stress tolerance to salt stress; The GmFVE protein is as follows (a) or (b) or (c): (a) the protein shown in Sequence 1 of the Sequence Listing; (b) a protein derived from soybean and having more than 98% identity and the same function as the protein shown in SEQ ID NO: 1 of the Sequence Listing; (c) A protein with the same function obtained by substituting and/or deleting and/or adding one or several amino acid residues to the protein shown in Sequence 1 of the sequence listing. 6. Application of GmFVE protein in regulating plant salt tolerance; The GmFVE protein is as follows (a) or (b) or (c): (a) the protein shown in Sequence 1 of the Sequence Listing; (b) a protein derived from soybean and having more than 98% identity and the same function as the protein shown in SEQ ID NO: 1 of the Sequence Listing; (c) A protein with the same function obtained by substituting and/or deleting and/or adding one or several amino acid residues to the protein shown in SEQ ID NO: 1 of the sequence listing. 7. the application of the biological material relevant with the GmFVE protein described in claim 6 in regulating and controlling the salt tolerance of plants; The biological material is any one of the following (d1) to (d12): (d1) a nucleic acid molecule encoding a GmFVE protein; (d2) an expression cassette containing the nucleic acid molecule of (d1); (d3) a recombinant vector containing the nucleic acid molecule of (d1); (d4) a recombinant vector containing the expression cassette of (d2); (d5) a recombinant microorganism containing the nucleic acid molecule of (d1); (d6) a recombinant microorganism containing the expression cassette of (d2); (d7) a recombinant microorganism containing the recombinant vector of (d3); (d8) a recombinant microorganism containing the recombinant vector of (d4); (d9) a transgenic plant cell line containing the nucleic acid molecule of (d1); (d10) a transgenic plant cell line containing the expression cassette of (d2); (d11) a transgenic plant cell line containing the recombinant vector of (d3); (d12) A transgenic plant cell line containing the recombinant vector of (d4). 8. application as claimed in claim 7, is characterized in that: Nucleic acid molecules encoding GmFVE proteins are as follows (e1), (e2) or (e3): (e1) a DNA molecule whose coding region is shown in Sequence 2 of the Sequence Listing; (e2) a DNA molecule having 75% or more identity with (e1) and encoding the GmFVE protein; (e3) a DNA molecule that hybridizes to (el) or (e2) under stringent conditions and encodes the GmFVE protein. 9. The application according to claim 7 or 8, wherein the regulation is negative regulation. 10. The use or method of any one of claims 1 to 9, wherein the plant is a monocotyledonous or dicotyledonous plant.

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