Application of TaHSP70 gene in regulation and control of plant salt tolerance

Abstract

The invention discloses application of a TaHSP70 gene in regulation and control of plant salt tolerance, and belongs to the technical field of molecular breeding. The nucleotide sequence of the TaHSP70 gene in the invention is as shown in SEQ ID NO: 1, SEQ ID NO: 2 and / or SEQ ID NO: 3. According to the invention, three TaHSP70 family genes are cloned from wheat, and the expression quantity of the three genes in wheat salt-stressed leaves is obviously improved. After the three TaHSP70 family genes in the wheat are mutated by adopting a Crispr / cas 9 method, the salt tolerance of the mutated wheat is obviously reduced. In conclusion, the three TaHSP70 family genes are related to the salt tolerance of the wheat, can be used for regulating and controlling the salt tolerance of the wheat and enriching wheat germplasm resources, and have important application value in breeding of the salt-tolerant wheat.

Description

Technical Field

[0001] The invention belongs to the technical field of molecular breeding, and specifically relates to the application of TaHSP70 gene in regulating plant salt tolerance. Background Art

[0002] As the degree of global soil salinization increases, the improvement of wheat yield and quality is severely restricted. Soil salinization can cause wheat growth retardation, and in severe cases can cause its death, and its yield and quality will also decrease. It is one of the important factors affecting agricultural production, greatly limiting the stable production and increase of grain crops, and seriously affecting grain production and agricultural sustainable development. Therefore, salt stress has become the main bottleneck for further improving its yield and quality.

[0003] Breeding salt-tolerant wheat varieties is of great significance for improving the yield and quality of wheat under salt stress. Traditionally, wheat varieties with salt tolerance and excellent comprehensive traits are bred through hybrid selection breeding, but the selection cycle is long and the workload is large. Using molecular biological methods to breed salt-tolerant wheat varieties can greatly reduce the workload of traditional methods and shorten the breeding cycle. Therefore, exploring the molecular mechanism of wheat salt tolerance and identifying the key genes using molecular biology will have important theoretical significance and practical value for breeding salt-tolerant varieties. Summary of the invention

[0004] In view of the problems existing in the prior art, the object of the present invention is to provide an application of the TaHSP70 gene in regulating plant salt tolerance.

[0005] In order to achieve the above object, the present invention adopts the following technical solution:

[0006] The application of TaHSP70 gene in regulating plant salt tolerance, the amino acid sequence of the TaHSP70 gene is shown in SEQ ID NO:4 and / or SEQ ID NO:5.

[0007] Based on the above scheme, the nucleic acid sequence of the TaHSP70 gene is shown as SEQ ID NO:1, SEQ ID NO:2 and / or SEQ ID NO:3.

[0008] On the basis of the above scheme, the salt tolerance of plants is reduced by mutating or silencing the TaHSP70 gene in plants; or the TaHSP70 gene in plants is overexpressed to improve the salt tolerance of plants.

[0009] Based on the above scheme, the plant is wheat.

[0010] A method for regulating plant salt tolerance, comprising: reducing plant salt tolerance by mutating or silencing a TaHSP70 gene in the plant; or overexpressing the TaHSP70 gene in the plant to improve plant salt tolerance; the amino acid sequence of the TaHSP70 gene is shown in SEQ ID NO:4 and / or SEQ ID NO:5.

[0011] Based on the above scheme, the nucleic acid sequence of the TaHSP70 gene is shown as SEQ ID NO:1, SEQ ID NO:2 and / or SEQ ID NO:3.

[0012] Based on the above scheme, the method of mutating the TaHSP70 gene in the plant is to use the Crispr / cas 9 method to mutate at least one sequence in SEQ ID NO:1-SEQ ID NO:3 in the plant.

[0013] Based on the above scheme, the mutation is the insertion, deletion or substitution of bases.

[0014] Based on the above scheme, the plant is wheat.

[0015] The use of the TaHSP70 gene in the preparation of a product for regulating plant salt tolerance, wherein the amino acid sequence of the TaHSP70 gene is shown in SEQ ID NO:4 and / or SEQ ID NO:5.

[0016] A product for regulating plant salt tolerance is a recombinant expression vector, an expression cassette, a recombinant bacterium, a recombinant virus or a transgenic cell line containing a TaHSP70 gene sequence; the amino acid sequence of the TaHSP70 gene is shown in SEQ ID NO:4 and / or SEQ ID NO:5.

[0017] Advantages of the technical solution of the present invention:

[0018] HSP70 (Heat shock 70kDa protein) is a heat shock protein with a molecular weight of about 70kDa. It is an important member of the heat shock protein family and is known as the major heat shock protein. It includes more than 20 proteins with molecular weights of 68, 72, 73, 75, 78kDa, etc.

[0019] The present invention clones and obtains three TaHSP70 family genes from wheat, namely TaHSP70-4, TaHSP70-10 and TaHSP70-14; the expression levels of these three genes in wheat leaves after salt stress are significantly increased. After the TaHSP70-4, TaHSP70-10 and TaHSP70-14 genes in wheat are mutated by Crispr / cas 9 method, the TaHSP70 Crispr stable strain is obtained. Under salt stress, the TaHSP70 Crispr stable strain wilts significantly and the leaves turn yellow seriously, the SOD and POD enzyme activities decrease, the MDA content increases, the seed width becomes narrower, and the grain length becomes shorter. It can be seen that the salt tolerance of wheat is reduced after the TaHSP70-4, TaHSP70-10 and TaHSP70-14 genes are mutated. In summary, the three TaHSP70 genes cloned in the present invention are related to the salt tolerance of wheat, can be used to regulate the salt tolerance of wheat, enrich the germplasm resources of wheat, and have important application value in salt-tolerant wheat breeding. BRIEF DESCRIPTION OF THE DRAWINGS

[0020] Figure 1 It is an amino acid sequence alignment diagram of TaHSP70A, TaHSP70B, and TaHSP70D;

[0021] Figure 2 The expression analysis of TaHSP70 gene in wheat leaves before and after salt stress;

[0022] Figure 3 The expression analysis of TaHSP70 gene in roots and leaves before and after salt stress (Figure A is the expression analysis of TaHSP70 gene in leaves; Figure B is the expression analysis of TaHSP70 gene in roots);

[0023] Figure 4 This is the electrophoresis diagram of the screening of TaHSP70 Crispr stable strains (wherein, lane M is a marker; lane 1 is the recipient variety CB037, lanes 2-23 are transgenic strains hsp70-1 to hsp70-22; water refers to the negative control);

[0024] Figure 5 are the mutation sites of TaHSP70A, TaHSP70B, and TaHSP70D in CB037 and Crispr stable strains;

[0025] Figure 6 is the salt-tolerant phenotype of the TaHSP70 Crispr stable strain (where A is the salt-tolerant phenotype of CB037 and Crispr stable strains under normal conditions; B is the salt-tolerant phenotype of CB037 and Crispr stable strains under salt stress; C is the relative plant height, leaf length and leaf width of CB037 and Crispr stable strains after salt stress);

[0026] Figure 7 The physiological indexes of TaHSP70 Crispr stable strains were measured (A is SOD activity after salt stress; B is POD activity after salt stress; C is MDA content after salt stress);

[0027] Figure 8 This is the grain phenotype of the TaHSP70 Crispr stable strain at maturity (where A is grain width and B is grain length). DETAILED DESCRIPTION

[0028] The terms used in the present invention, unless otherwise specified, generally have the meanings commonly understood by those of ordinary skill in the art. The present invention will be further described in detail below in conjunction with specific examples and with reference to data. The following examples are intended to illustrate the present invention and are not intended to limit the scope of the present invention in any way.

[0029] The experimental methods in the following examples, unless otherwise specified, are all conventional methods, and are performed according to the techniques or conditions described in the literature in the field or according to the product instructions. The experimental materials, reagents, drugs, etc. used in the following examples, unless otherwise specified, can all be purchased through general channels.

[0030] Chinese Spring (CS) wheat seeds are maintained by the Wheat Genetics and Breeding Laboratory, College of Agriculture, Qingdao Agricultural University.

[0031] Example 1

[0032] Cloning of TaHSP70 family genes

[0033] Using the leaves of wheat variety Chinese Spring as materials, total RNA was extracted, reverse transcribed into cDNA as a template, and PCR amplification was performed using specific primers of the TaHSP70 gene to obtain three TaHSP70 family gene sequences, namely TaHSP70-4, TaHSP70-10 and TaHSP70-14, whose nucleic acid sequences are shown in SEQ ID NO: 1, SEQ ID NO: 2, and SEQ ID NO: 3, respectively. For the convenience of expression, TaHSP70-4, TaHSP70-10 and TaHSP70-14 are represented by TaHSP70A, TaHSP70B and TaHSP70D, respectively, in the following text and in the accompanying drawings, the amino acid sequences encoded by TaHSP70-4 and TaHSP70-10 are shown in SEQ ID NO: 4, and the amino acid sequence encoded by TaHSP70-14 is shown in SEQ ID NO: 5. Unlike TaHSP70-4 and TaHSP70-10, the amino acid at position 637 of the amino acid sequence of TaHSP70-14 changes from A to G ( Figure 1 ).

[0034] SEQ ID NO:1(5’→3’)

[0035]

[0036] SEQ ID NO:2(5’→3’)

[0037]

[0038] SEQ ID NO:3(5’→3’)

[0039]

[0040] SEQ ID NO:4

[0041] *

[0042] SEQ ID NO:5

[0043] MAKGEGPAIGIDLGTTYSCVGVWQHDRVEIIANDQGNRTTPSYVAFTDTERLIGDAAKNQVAMNPTNTVFDAKRLIGRRFSDPSVQSDMKLWPFKVIPGPADKPMIVVNYKGEEKQFAAEEISSMVLIKMREIAEAFLGNSVKNAVVTVPAYFNDSQRQATKDAGAIAGLNVLRIINEPTAAAIAYGLDKKATSTGEKNVLIFDLGGGTFDVSLLTIEEGIFEVKATAGDTHLGGEDFDNRMVNHFVQEFKRKHKKDITGNPRALRRLRTACERAKRTLSSTAQTTIEIDSLYEGVDFYTTITRARFEELNMDLFRKCMEPVEKCLRDAKMDKSTVHDVVLVGGSTRIPKVQQLLQDFFNGKELCKSINPDEAVAYGASVQAAILSGEGNEKVQDLLLLDVTPLSLGLETAGGVMTTLIPRNTTIPTKKEQVFSTYSDNQPGVLIQVYEGERARTKDNNLLGKFELSGIPPAPRGVPQITVCFDIDANGILNVSAEDKTTGQKNKITITNDKGRLSKEEIEKMVQEAERYKAEDEEHKKKVDAKNALENYAYNMRNTVKDDKIASKLGADDKKKVEEAIEGTISWLDANQLAEADEFEDKMKELEGICNPIIAKMYQGAAPDMGGGMGMDEDMPAGGGGAGPKIEEVD*

[0044] The specific primer sequences of the TaHSP70 gene are as follows:

[0045] TaHSP70-F: 5'-ATGGCCAAGGGCGAGGGG-3' (SEQ ID NO:6);

[0046] TaHSP70-R: 5'-TTAGTCCACCTCCTCGATCTT-3' (SEQ ID NO:7);

[0047] PCR reaction system (25 μL): 8 μL of ddH2O, 12.5 μL of 2×Phanta Max Master Mix, 1 μL of each upstream and downstream primer, and 2.5 μL of cDNA template.

[0048] PCR reaction conditions:

[0049] Pre-denaturation at 95°C for 3 min; denaturation at 95°C for 15 s, annealing at 59°C for 15 s, extension at 72°C for 2 min (1 kb / min), 35 cycles; further extension at 72°C for 5 min; storage at 4°C.

[0050] Example 2

[0051] Expression analysis of TaHSP70 family genes in wheat leaves before and after salt stress

[0052] The Chinese spring was hydroponically cultured until the one-leaf-one-heart stage, and treated with 250mmol / L NaCl solution for 0h, 1h, 3h, 6h, 12h, and 24h, and the leaves were taken to extract total RNA using the Trizol method, and the final concentration of RNA was controlled at about 1000ng / μL for reverse transcription. The first-strand cDNA synthesis kit used was the Hiscript kit from Nanjing Novozyme Biotechnology Co., Ltd. RT SuperMix for qPCR (+gDNA wiper). Using the reverse transcribed cDNA as a template, the ChamQTMSYBRColor qPCR Master Mix (Vazyme) kit, the expression level of the wheat TaActin gene as an internal reference, and the fluorescent quantitative PCR primers of the TaHSP70 gene were used to analyze the expression pattern of the TaHSP70 family genes at different time points after salt stress.

[0053] The comparative threshold method is used to quantitatively analyze the results of real-time quantitative PCR, set the fluorescence threshold, and determine the cycle number Ct value under the fluorescence threshold. According to the Ct value, calculate the C value. C = 2 -△Ct , △Ct=Ct 目的基因 -C t内参基因 The average of the three repeated C values ​​was calculated as the relative expression level of the target gene.

[0054] Reaction system (10 μL): 2×ChamQ SYBR Color qPCR Master Mix 5 μL, 50×ROX Reference Dye II 0.2 μL, Forward Primer (10 μM) 0.2 μL, Reverse Primer (10 μM) 0.2 μL, 3-fold diluted first-strand cDNA solution 0.5 μL, ddH2O 3.9 μL.

[0055] The qRT-PCR amplification procedure uses a three-step approach:

[0056] Step 1: pre-denaturation at 95°C for 3 min;

[0057] Step 2: PCR reaction: 95°C for 15 s, 59°C for 20 s, 2°C for 20 s, for a total of 40 cycles;

[0058] Step 3: Melting curve: 95℃1s, 60℃20s, 95℃1s.

[0059] The fluorescent quantitative PCR primers of the TaHSP70 gene are as follows:

[0060] TaActin-qRT-F: 5'-TATGCCAGCGGTCGAACAAC-3' (SEQ ID NO: 8)

[0061] TaActin-qRT-R: 5'-GGAACAGCACCTCAGGGCAC-3' (SEQ ID NO: 9)

[0062] TaHSP70-qRT-AL:5'-GAGGAGGTGGACTAAGCTGTTC-3'(SEQ ID NO:10)

[0063] TaHSP70-qRT-AR: 5'-CACGACGACAGACGACTGG-3' (SEQ ID NO: 11)

[0064] TaHSP70-qRT-BL: 5'-CGACAAGAAGAAGGTGGAGGAC-3' (SEQ ID NO: 12)

[0065] TaHSP70-qRT-BR:5'-GGCATCGAAAGGACACCC-3'(SEQ ID NO:13)

[0066] TaHSP70-qRT-DL: 5'-CGCGCTACTTGATGGGTTC-3'(SEQ ID NO:14)

[0067] TaHSP70-qRT-DR: 5'-CCAACGACTCCATCGAACG-3' (SEQ ID NO: 15)

[0068] The results showed that the expression levels of TaHSP70A, TaHSP70B, and TaHSP70D genes were the highest 1 h after salt stress, and the expression level of TaHSP70D was significantly higher than that of TaHSP70A and TaHSP70B, indicating that TaHSP70A, TaHSP70B, and TaHSP70D were all involved in the response to salt stress in wheat, among which TaHSP70D played a more important role ( Figure 2 ).

[0069] Example 3

[0070] Expression analysis of three genes of TaHSP70 family in wheat leaves and roots before and after salt stress

[0071] Chinese spring was hydroponically cultured to the one-leaf-one-heart stage, treated with 250mmol / L NaCl solution for 1h, and the leaves and roots before and after salt stress were taken as materials for real-time fluorescence quantitative analysis. The method was as in Example 2. The results showed that the expression levels of TaHSP70A, TaHSP70B, and TaHSP70D genes were all upregulated in the leaves after salt stress, and TaHSP70D had the highest expression level, but their expression levels were all downregulated in the roots ( Figure 3 ). Therefore, all three genes of the TaHSP70 family are involved in the salt stress response of wheat leaves, and the TaHSP70D gene has a stronger response than TaHSP70A and TaHSP70B.

[0072] Example 4

[0073] Functional identification of TaHSP70 family genes

[0074] 1. Determination of TaHSP70 CRISPR stable strains and mutation types

[0075] The TaHSP70 Crispr strain was constructed by Crispr / cas 9 technology. The receptor variety CB037 and Crispr strain DNA were extracted by CTAB method. First, Cas 9 primers were used to detect whether the T2 generation TaHSP70 Crispr strain still had Cas 9 protein. The detection primers are as follows:

[0076] Cas 9-F: 5'-CTCACAAATTATCAGCACGCTAGTC-3' (SEQ ID NO: 16);

[0077] Cas 9-R: 5'-TCACCTTAGTCAGCCCGTTG-3' (SEQ ID NO: 17);

[0078] Then, the mutation sites of TaHSP70A, TaHSP70B, and TaHSP70D genes were detected for the Crispr strains that did not have Cas 9 protein to determine their mutation types. The mutation detection primers are as follows:

[0079] TaHSP70AL: 5'-GCGAGGAGAAGCAGTTTGCT-3' (SEQ ID NO: 18);

[0080] TaHSP70AR: 5'-TCTCAATGGTGGTCTGTGCA-3' (SEQ ID NO: 19);

[0081] TaHSP70BL: 5'-ATTGGCAGGAGGTTCTCTGAT-3' (SEQ ID NO: 20);

[0082] TaHSP70BR: 5'-GCGGTTGTCGAAGTCCTCT-3' (SEQ ID NO: 21);

[0083] TaHSP70DL: 5'-GTTCTGTTCCTCTGTTTCTGCTTA-3' (SEQ ID NO: 22);

[0084] TaHSP70DR: 5'-CGAGCCCGTGTGATGGTA-3' (SEQ ID NO: 23);

[0085] Using the leaf DNA of the T2 generation TaHSP70 Crispr strain as material, PCR amplification was performed using Cas 9 primers, and four Crispr strains without Cas 9 protein were identified, which were named hsp70-1, hsp70-2, hsp70-3 and hsp70-4 ( Figure 4 ). The four Crispr strains without Cas 9 protein were used as research materials, and primers for detecting mutation sites of TaHSP70A, TaHSP70B, and TaHSP70D were designed outside the left and right targets. The mutation sites of these four wheat strains were analyzed, and the results showed that ( Figure 5 ), the TaHSP70A gene in hsp70-1 and hsp70-2 lacks 6 bases at the left target site;

[0086] The left target site of TaHSP70A gene in hsp70-3 and hsp70-4 lacked 7 and 2 bases, respectively; the first base of the right target site of TaHSP70B gene in hsp70-1 mutated from G to T; the right target sites of TaHSP70A and TaHSP70D genes in hsp70-2 lacked 2 and 4 bases, respectively; the right target sites of TaHSP70A, TaHSP70B and TaHSP70D genes in hsp70-3 lacked 6, 2 and 8 bases, respectively, and the first base of the right target site of TaHSP70B gene mutated from G to T; the right target sites of TaHSP70A, TaHSP70B and TaHSP70D genes in hsp70-4 lacked 2, 2 and 8 bases, respectively, and the first base of the right target site of TaHSP70B gene also mutated from G to T. In summary, the results showed that hsp70-1 was a double mutation type of A and B, hsp70-2 was a double mutation type of A and D, and hsp70-3 and hsp70-4 were triple mutation types of A, B and D.

[0087] 2. Phenotypic identification of salt-tolerant strains of TaHSP70 CRISPR stable strains

[0088] 25 seeds of CB037 with the same size and 4 Crispr strains (hsp70-1, hsp70-2, hsp70-3 and hsp70-4) without Cas 9 protein after mutation detection were selected, disinfected with 1% NaClO for 10 minutes, rinsed with distilled water 4-5 times, placed in a culture dish and cultured for 3 days, and then transplanted into a 20 cm diameter flower pot for soil culture. Two treatments and two replicates were set for each variety. After transplanting, they were treated with 1.5 mol / L NaCl solution for seedling phenotypic identification.

[0089] The results showed that compared with the control CB037, after salt stress, the TaHSP70 Crispr strains hsp70-1, hsp70-2, hsp70-3 and hsp70-4 wilted significantly and their leaves turned yellow severely; among them, the phenotypes of the A, B, D triple mutant wheat strains hsp70-3 and hsp70-4 were more obvious than those of the A, B double mutant wheat strains hsp70-1 and the A, D double mutant wheat strains hsp70-2, indicating that the functions of the three partial homologous genes A, B, and D have an additive effect ( Figure 6 A and B in ).

[0090] The statistical results showed that after salt stress treatment, the plant height, leaf length and leaf width of the four TaHSP70 Crispr lines were smaller than those of the control variety CB037 ( Figure 6 C).

[0091] 3. Determination of physiological indicators

[0092] After the salt stress treatment phenotypes of CB037 and Crispr strains (hsp70-1, hsp70-2, hsp70-3 and hsp70-4) emerge, take their leaves and measure the physiological indicators SOD, POD and MDA. The measurement method is as follows:

[0093] SOD enzyme activity assay

[0094] (1) Weigh 0.2 g of wheat leaves in a pre-cooled mortar, add 1 mL of pre-cooled extract (the extract is PBS sodium phosphate buffer), grind until homogenized, 4°C, 10,000 r, 15 min, and take the supernatant as the crude SOD extract;

[0095] (2) Add 1.5 mL of 0.05 mol / L PBS solution, 0.3 mL of 130 mmol / L Met solution, 750 μmol / L NBT solution, 20 μmol / L riboflavin solution, and 100 μmol / L EDTA-Na2 solution to the control tube and the assay tube in sequence. Add 0.1 mL of enzyme solution to the assay tube. Do not add enzyme solution to the control tube, but add 0.1 mL of buffer solution instead. Fill the total volume to 3.3 mL with distilled water.

[0096] (3) Set up three replicates, shield one of the control tubes from light and place it under a 4,000-lx fluorescent light for 20-30 min at 25-35°C, then compare the color at 560 nm.

[0097] The SOD enzyme activity was calculated according to the following formula:

[0098]

[0099] Ao: light absorption value of the illuminated control tube; As: light absorption value of the sample tube; Vt: total volume of sample solution (mL); V1: amount of sample used during measurement (mL); FW: fresh weight of sample (g).

[0100] POD enzyme activity assay

[0101] (1) Weigh 0.25 g of leaves, add 5 times the amount of (M / V) pH 7.0 PBS, grind on ice at 15000 r for 15 min, and the supernatant is the crude POD extract;

[0102] (2) Add 1 mL of 0.3% H2O2, 0.95 mL of 0.2% guaiacol, and 1 mL of PBS at pH 7.0 to the reaction tube in sequence, and finally add 0.05 mL of enzyme solution to start the reaction, and record the rate of increase of OD at 470 nm. An increase of 0.01 OD per minute is defined as 1 activity unit; calculate the POD enzyme activity according to the following formula:

[0103]

[0104] ΔA 470 : Change in absorbance during reaction time; W: plant fresh weight (g); Vt: total volume of extracted enzyme solution (mL); Vs: volume of enzyme solution used in determination (mL); t: reaction time, min.

[0105] Determination of MDA content

[0106] (1) Weigh 1 g of wheat leaves, add 2 mL of 10% TCA and grind in a mortar, then add 8 mL of TCA and continue grinding until homogenized, at 4000 r / min for 10 min. The supernatant is the crude MDA extract.

[0107] (2) Take 2 mL of the supernatant, add 2 mL of 0.6% TBA, mix well, react in boiling water for 15 min, take the supernatant, and measure the absorbance at 450, 532 and 600 nm; calculate the concentration and content of MDA according to the following formula:

[0108] MDA concentration = 6.45*(D 532 -D 600 )-0.56*D 450

[0109] MDA content = 2MDA concentration*extract volume (ml) / fresh weight (g) / 1000.

[0110] The results showed that after salt stress, the SOD and POD activities of the TaHSP70 Crispr stable strains decreased, and the MDA content increased. Among them, hsp70-3 and hsp70-4 changed most significantly, indicating that these two strains were more sensitive to salt stress than hsp70-1 and hsp70-2 ( Figure 7 ). This is consistent with the results of seedling phenotype identification of TaHSP70 Crispr stable strain.

[0111] 4. Identification of grain phenotypes of TaHSP70 Crispr stable strains at maturity

[0112] The seeds of CB037 and TaHSP70 Crispr stable strains (hsp70-1, hsp70-2, hsp70-3 and hsp70-4) were harvested and the grain phenotypes were identified at maturity. The results showed that after salt stress, the seeds of TaHSP70 Crispr strains became narrower in width and shorter in length ( Figure 8 ).

[0113] The above is only a preferred embodiment of the present invention, and does not limit the present invention in other forms. Any technician familiar with the profession may use the above disclosed technical content to change or modify it into an equivalent embodiment with equivalent changes. However, any simple modification, equivalent change and modification made to the above embodiment according to the technical essence of the present invention without departing from the technical solution of the present invention still belongs to the protection scope of the technical solution of the present invention.

Claims

1. Application of TaHSP70 gene in regulating plant salt tolerance, characterized in that: The amino acid sequence of the TaHSP70 gene is shown in SEQ ID NO:4 and / or SEQ ID NO:

5.

2. The use of the TaHSP70 gene in regulating plant salt tolerance according to claim 1, characterized in that: The nucleic acid sequence of the TaHSP70 gene is shown in SEQ ID NO:1, SEQ ID NO:2 and / or SEQ ID NO:

3.

3. The use of the TaHSP70 gene in regulating plant salt tolerance according to claim 1 or 2, characterized in that: The salt tolerance of plants can be reduced by mutating or silencing the TaHSP70 gene in plants; or the salt tolerance of plants can be improved by overexpressing the TaHSP70 gene in plants.

4. The use of the TaHSP70 gene in regulating plant salt tolerance according to claim 3, characterized in that: The plant is wheat.

5. A method for regulating plant salt tolerance, characterized in that: The salt tolerance of the plant is reduced by mutating or silencing the TaHSP70 gene in the plant; or the TaHSP70 gene in the plant is overexpressed to improve the salt tolerance of the plant; the amino acid sequence of the TaHSP70 gene is shown in SEQ ID NO:4 and / or SEQ ID NO:

5.

6. The method for regulating plant salt tolerance according to claim 5, characterized in that: The nucleic acid sequence of the TaHSP70 gene is shown in SEQ ID NO:1, SEQ ID NO:2 and / or SEQ ID NO:

3.

7. The method for regulating plant salt tolerance according to claim 6, characterized in that: The method of mutating the TaHSP70 gene in the plant is to use the Crispr / cas 9 method to mutate at least one sequence of SEQ ID NO:1-SEQ ID NO:3 in the plant.

8. The method for regulating plant salt tolerance according to claim 7, characterized in that: The mutation is insertion, deletion or substitution of bases.

9. The method for regulating plant salt tolerance according to any one of claims 6 to 8, characterized in that: The plant is wheat.

10. Application of TaHSP70 gene in the preparation of a product for regulating plant salt tolerance, characterized in that: The amino acid sequence of the TaHSP70 gene is shown in SEQ ID NO:4 and / or SEQ ID NO:5.

Images