Application of wheat gene tagamyb-like in improving plant salt tolerance

Abstract

The application discloses application of a wheat gene TaGAMYB-like in improving plant salt tolerance and belongs to the technical field of functional genomics. The nucleotide sequence of the wheat gene TaGAMYB-like is shown in SEQ ID NO:1, and the gene can improve the salt stress tolerance of plants. The application has the advantages that the wheat gene TaGAMYB-like is transformed into a model plant Arabidopsis thaliana, and the salt tolerance of the Arabidopsis thaliana can be obviously improved. If the wheat gene TaGAMYB-like is transformed into plants such as wheat, rice, corn and cauliflower, new germplasm with salt tolerance characteristics can be obtained, and the application has important significance for the cultivation of excellent crop new varieties and the wide application of the excellent crop new varieties in production.

Description

TECHNICAL FIELD

[0001] The application relates to application of a wheat gene, in particular to application of a wheat gene TaGAMYB-like in improving plant salt tolerance, and belongs to the technical field of functional genomics. BACKGROUND

[0002] Salt stress is a serious adverse environmental factor, which can cause osmotic stress, cell ion toxicity and oxidative stress on crops, and further causes seed germination inhibition, photosynthetic performance decline, growth and development stagnation and the like, and seriously affects crop yield. Soil salinization has become one of important adverse factors affecting wheat growth and yield increase. Therefore, improving the salt tolerance of wheat is one of important ways to ensure food security, and cultivating salt-tolerant varieties is an effective measure to improve the salt tolerance of wheat, and further mining and identifying key salt-tolerant genes is a scientific and reasonable biological technology means for cultivating salt-tolerant varieties. SUMMARY

[0003] The application screens and clones a wheat gene TaGAMYB-like from Chinese spring (CS) wheat, and proves that the gene can effectively improve the salt tolerance of Arabidopsis.

[0004] In order to achieve the above target, the application adopts the following technical scheme:

[0005] The application of the wheat gene TaGAMYB-like in improving plant salt tolerance, the nucleotide sequence of the wheat gene TaGAMYB-like is shown as SEQ ID NO: 1, and the gene can improve the salt stress tolerance of plants.

[0006] Preferably, the aforementioned application comprises the following steps:

[0007] (1) PCR amplification and cloning of the wheat gene TaGAMYB-like;

[0008] (2) connecting the cloned wheat gene TaGAMYB-like into an expression vector to obtain an overexpression recombinant vector;

[0009] (3) transforming the overexpression recombinant vector into agrobacterium to obtain an overexpression recombinant strain;

[0010] (4) transforming the overexpression recombinant strain into plants, and screening and obtaining an overexpression line of the wheat gene TaGAMYB-like.

[0011] More preferably, the aforementioned expression vector is super1300 (GFP-C), and the aforementioned agrobacterium is Agrobacterium tumefaciens GV3101.

[0012] The wheat gene TaGAMYB-like is transformed into the model plant Arabidopsis thaliana, and the salt tolerance of the Arabidopsis thaliana is obviously improved; the wheat gene TaGAMYB-like is transformed into wheat, rice, corn, cauliflower and other plants, and new germplasm with salt tolerance characteristics can be obtained, which has important significance for the cultivation of excellent crop new varieties and the wide application of excellent crop new varieties in production. BRIEF DESCRIPTION OF DRAWINGS

[0013] Figure 1 is a structural schematic diagram of the target vector super1300 (GFP-C);

[0014] Figure 2 is a PCR identification diagram of the TaGAMYB-like overexpression transgenic Arabidopsis thaliana strain (OE1, OE2);

[0015] Figure 3 is a relative expression amount statistical diagram of the gene TaGAMYB-like of the TaGAMYB-like overexpression transgenic Arabidopsis thaliana strain (OE1, OE2) and the wild type (WT) of the Columbia ecotype Arabidopsis thaliana;

[0016] Figure 4 is a germination rate statistical diagram of the TaGAMYB-like overexpression transgenic Arabidopsis thaliana seed (OE1, OE2) and the wild type Arabidopsis thaliana seed (WT) of the Columbia ecotype Arabidopsis thaliana after ABA treatment;

[0017] Figure 5 is a germination rate statistical diagram of the TaGAMYB-like overexpression transgenic Arabidopsis thaliana seed (OE1, OE2) and the wild type Arabidopsis thaliana seed (WT) of the Columbia ecotype Arabidopsis thaliana after salt stress treatment;

[0018] Figure 6 is a comparison diagram of the root growth of each strain under normal conditions (control group);

[0019] Figure 7 is a root length statistical diagram of each strain under normal conditions (control group);

[0020] Figure 8 is a comparison diagram of the root growth of each strain under salt stress treatment conditions;

[0021] Figure 9 is a root length statistical diagram of each strain under salt stress treatment conditions;

[0022] Figure 10 is a plant height statistical diagram of each strain under normal conditions and under salt stress treatment conditions;

[0023] Figure 11Figure 3 is a fresh weight statistical graph of each strain under normal conditions and under salt stress treatment conditions;

[0024] Figure 12 Figure 4 is a dry weight statistical graph of each strain under normal conditions and under salt stress treatment conditions;

[0025] Figure 13 Figure 5 is a relative conductivity statistical graph of each strain under normal conditions and under salt stress treatment conditions;

[0026] Figure 14 Figure 6 is a chlorophyll content statistical graph of each strain under normal conditions and under salt stress treatment conditions;

[0027] Figure 15 Figure 7 is a SOD activity statistical graph of each strain under normal conditions and under salt stress treatment conditions;

[0028] Figure 16 Figure 8 is a POD activity statistical graph of each strain under normal conditions and under salt stress treatment conditions. DETAILED DESCRIPTION

[0029] The present application will be described in detail below in conjunction with the accompanying drawings and examples. The reagents, materials and instruments used in the examples can be obtained from commercial channels unless otherwise specified. The terms used in the examples have the meanings generally understood by those of ordinary skill in the art unless otherwise specified. The experimental methods used in the examples are conventional methods unless otherwise specified.

[0030] Example 1: Cloning of wheat gene TaGAMYB-like

[0031] Wheat is an important food crop with a wide planting range, and is also a crop with strong adaptability to harsh external environments. The present application attempts to screen salt-tolerant genes from wheat, and ultimately screens a salt-tolerant gene from Chinese Spring (CS) wheat. In the Ensembl Plants database, the gene number of the gene is TraesCS1A02G308100.2, which is located on wheat chromosome 1A, the ORF (open reading frame) is 1380 bp, the mRNA length is 2244 bp, the nucleotide sequence is shown as SEQ ID NO: 1, the size of the encoded product is 459 amino acids, the molecular weight is 50578.18, and the isoelectric point is 5.12. The gene is named TaGAMYB-like, and it is cloned by cloning technology, and the steps are as follows:

[0032] Chinese Spring (CS) wheat was taken and total RNA was extracted from leaves by Trizol method after 7 days of water culture. After purification, cDNA was obtained by reverse transcription using HiScript III RT SuperMix for qPCR (+gDNA wiper) reverse transcriptase. The cDNA was used as a template, and the upstream primer L1 shown in SEQ ID NO: 3 and the downstream primer R1 shown in SEQ ID NO: 4 were used to amplify the gene TaGAMYB-like. The amplified product was sequenced to determine the nucleotide sequence of the target gene.

[0033] The nucleotide sequences of the upstream primer L1 and the downstream primer R1 are as follows:

[0034] The upstream primer L1 is 5'-ATGACTCGGGCTAAGAGC-3' (SEQ ID NO: 3);

[0035] The downstream primer R1 is 5'-CTGGAAGATGGGCGGCAT-3' (SEQ ID NO: 4).

[0036] The PCR reaction system for amplifying the gene TaGAMYB-like is as follows: ddH2O 8 μL, 2×Phanta Max MasterMix (Dye Plus) 12.5 μL, upstream primer L1 1 μL, downstream primer R1 1 μL, and cDNA template 2.5 μL.

[0037] The PCR reaction conditions for amplifying the gene TaGAMYB-like are as follows: 95℃, 3 min; 95℃, 15 s, 59℃, 15 s, 72℃, 30 s, repeat 35 cycles; 72℃, extend for 5 min, and finally store at 4℃.

[0038] The obtained PCR amplification product was electrophoresed in a 1% agarose gel, and the fragment with a sequence length of about 1377 bp was recovered. Then, 5 min TM TA / Blunt-Zero Cloning Kit was connected to pCE2 TA / Blunt-Zero, and Escherichia coli Fast-T1 competent cells were transformed. Positive clones were selected for sequencing.

[0039] The sequencing results showed that a DNA fragment with a sequence length of 1377 bp was inserted into the pCE2 TA / Blunt-Zero vector. The nucleotide sequence of the DNA fragment is shown in SEQ ID NO: 2, which is consistent with the nucleotide sequence of the gene with gene number TraesCS1A02G308100.2 in the Ensembl Plants database without a stop codon, and it is determined to be the full-length ORF sequence of the wheat gene TaGAMYB-like.

[0040] Example 2: Construction of a recombinant expression vector

[0041] 1. Amplification of the target gene

[0042] XbaI restriction sites were added to the 5' ends of the upstream primer L1 and the downstream primer R1 to obtain the upstream primer L2 and the downstream primer R2. The nucleotide sequences of the upstream primer L2 and the downstream primer R2 are shown as follows, respectively:

[0043] Upstream primer L2:

[0044] 5'-atacaccaaatcgactctagaATGACTCGGGCTAAGAGCGAG-3' (SEQ ID NO: 5);

[0045] Downstream primer R2:

[0046] 5'-cataggtacccgggctctagaCTGGAAGATGGGCGGCAT-3' (SEQ ID NO: 6).

[0047] The positive E. coli liquid containing the pCE2 TA / Blunt-Zero recombinant vector obtained in Example 1 was used as a template for PCR amplification.

[0048] The PCR reaction system was as follows: ddH2O 8 μL, 2x Phanta Max Master Mix (Dye Plus) 12.5 μL, upstream primer L2 1 μL, downstream primer R2 1 μL, and template 2.5 μL.

[0049] The PCR reaction program was as follows: 95°C, 3 min; 95°C, 15 s, 59°C, 15 s, 72°C, 30 s, repeated for 35 cycles; 72°C, 5 min for extension, and finally 4°C storage.

[0050] The PCR product (target gene) with a sequence length of about 1377 bp was recovered for subsequent reactions.

[0051] 2. Linearization of the vector

[0052] QuickCut TM The target vector super1300 (GFP-C) was linearized by XbaI restriction endonuclease, and the structure of the target vector super1300 (GFP-C) is shown in Figure 1 , to obtain a linearized cloning vector.

[0053] Enzyme digestion system: 1 μg of plasmid, 2 μL of 10×QuickCut Green Buffer, 1 μL of QuickCut XbaI, and ddH2O to 20 μL. TM XbaI 1 μL, and ddH2O to 20 μL.

[0054] Reaction condition: 37°C incubation for 3 h.

[0055] 3. Ligation of the target gene and the cloning vector

[0056] The target gene obtained in step 1 was connected to the linearized cloning vector obtained in step 2 by using a CloneExpress II One step Cloning Kit (Vayme) to obtain a recombinant expression vector super1300 (GFP-C)-TaGAMYB-like.

[0057] The ligation system was as follows: 3 μL of linearized super1300 (GFP-C), 1 μL of the target gene, 2 μL of 5×CE II Buffer, 1 μL of Exnase II, and 3 μL of ddH2O.

[0058] Reaction condition: the ligation system was mixed by gently blowing with a pipette, the liquid was collected at the bottom of the tube by brief centrifugation, and the reaction was performed at 37°C for 30 min.

[0059] 4. Screening and identification of the cloned target gene

[0060] The screening and identification of the cloned target gene were as follows:

[0061] (1) 10 μL of the recombinant reaction product obtained in step 3 was used to transform the competent cells of Escherichia coli Fast-T1, which was incubated at 37°C for 12 h (the culture medium was LB solid medium with the addition of 50 μg / mL kanamycin sulfate);

[0062] (2) The single colony obtained in step (1) was selected and incubated at 37°C and 180 rpm for 12 h (the culture medium was LB liquid medium with the addition of 50 μg / mL kanamycin sulfate);

[0063] (3) The bacterial solution obtained in step (2) was used as a template, and PCR identification was performed by using the upstream primer L1 and the downstream primer R1;

[0064] (4) The bacterial solution with positive PCR identification in step (3) was sent to GenScript Biotech (Shanghai) Co., Ltd. for sequencing.

[0065] The sequencing result of the TaGAMYB-like construct vector is shown in SEQ ID NO: 7. The reference sequence of the TaGAMYB-like construct vector is shown in SEQ ID NO: 8. The sequencing result shows that the recombinant expression vector containing the target gene (TaGAMYB-like), i.e., the recombinant expression vector super1300 (GFP-C)-TaGAMYB-like, is obtained, and the recombinant expression vector super1300 (GFP-C)-TaGAMYB-like is a linearized vector super1300 (GFP-C) to which the TaGAMYB-like sequence is added at the XbaI site.

[0066] The plasmid of the bacterial liquid with correct sequencing is extracted and stored at -20°C, and used for subsequent Agrobacterium transformation experiments.

[0067] Example 3: Transformation of Arabidopsis thaliana with the wheat gene TaGAMYB-like

[0068] 1. Construction of recombinant Agrobacterium

[0069] The recombinant expression vector super1300 (GFP-C)-TaGAMYB-like prepared in Example 2 is used to transform the competent cells of Agrobacterium tumefaciens GV3101, and the positive monoclonal is selected and cultured in LB solid medium containing 50 μg / mL kanamycin sulfate and 20 μg / mL rifampicin at 28°C.

[0070] The positive monoclonal is selected and cultured at 28°C and 180 rpm for 12 h (the culture medium is LB liquid medium, and 50 μg / mL kanamycin sulfate and 20 μg / mL rifampicin are added).

[0071] 1 μL of the bacterial liquid is aspirated, and PCR identification is performed using the upstream primer L1 and the downstream primer R1.

[0072] The bacterial liquid that is positive in PCR identification is the recombinant Agrobacterium tumefaciens containing the recombinant expression vector super1300 (GFP-C)-TaGAMYB-like, and the recombinant Agrobacterium tumefaciens is named GV3101 / super1300 (GFP-C)-TaGAMYB-like.

[0073] 2. Obtaining of transgenic Arabidopsis thaliana

[0074] The method for obtaining of transgenic Arabidopsis thaliana is as follows:

[0075] (1) Plant preparation before infection: Wild-type seeds of Arabidopsis thaliana ecotype Columbia were vernalized at 4°C for 72 h, sowed in MS solid medium, and cultured at 22°C, 15 h light / 9 h dark, and humidity of 60%-70% in a culture room. When two true leaves were grown, the plants were transplanted into planting pots containing mixed culture medium (nutrient soil and vermiculite mixed at a mass ratio of 3:1). After the plants flowered, the top of the main branch was cut off (to promote the development of lateral branches), and sufficient water was poured the day before infection;

[0076] (2) Activation of Agrobacterium: 2 mL of recombinant Agrobacterium tumefaciens GV3101 / super1300 (GFP-C) - TaGAMYB-like bacterial solution was poured into a sterilized 250 mL conical flask in a clean bench, 150 mL of LB liquid medium containing 50 μg / mL kanamycin sulfate and 20 μg / mL rifampicin was added, and the mixture was cultured at 28°C in a full-temperature shaker for 16 h, until the OD 600 reached 0.8. The bacterial solution was divided into three 50 mL centrifuge tubes, centrifuged at 5500 g for 20 min, and the supernatant was discarded, leaving the bacterial cells;

[0077] (3) Preparation of resuspension solution: 2.5 g of sucrose was dissolved in 50 mL of distilled water, and 10 μL of silmet-77 was added;

[0078] (4) The resuspension solution was added to the activated Agrobacterium bacterial cells in batches, 10 mL each time, and the OD 600 was measured until it reached 0.8, obtaining the resuspended bacterial solution;

[0079] (5) Dipping flower method to infect Arabidopsis thaliana: at 9 o'clock in the morning (when flowering is most vigorous, which is conducive to infection), 1 mL of mixed resuspended bacterial solution was taken with a pipette gun head each time and dropped on the inflorescences of Arabidopsis thaliana for dipping, and after the inflorescences were completely dipped, dark treatment was immediately performed for 1 d, and water was poured during light. The plants were infected twice a week;

[0080] (6) Harvesting of seeds from infected Arabidopsis thaliana plants: the T0 generation plants treated by the second dipping were cultured to fruiting using a conventional method, and mature T0 generation seeds were harvested;

[0081] (7) Cultivation of T0 generation positive seedlings: the T0 generation seeds were disinfected and then uniformly spread on MS solid medium containing 60 μL / 100 mL of hygromycin. The medium was incubated until Arabidopsis thaliana seedlings grew true leaves, and some seedlings had healthy true leaves and longer roots (about 14 d). T0 generation positive seedlings were obtained and moved to planting pots containing mixed culture medium (nutrient soil and vermiculite mixed at a mass ratio of 3:1) for cultivation. The seeds harvested from a single plant were T1 generation;

[0082] (8) The T1 generation seeds containing hygromycin resistance were cultured into T1 generation lines according to the method described in step (7), if the ratio of positive seedlings to dead seedlings was about 3:1, it was a single copy line, then the plants in the planting pot were cultured to bear seeds by using conventional method, and the mature T2 generation seeds on each single plant of the T1 generation line were harvested;

[0083] (9) 10 T2 generation line seeds were randomly selected and subjected to hygromycin resistance screening according to the same method, and no hygromycin resistance separation was obtained, which was a homozygous line, and finally two T2 generation homozygous lines of TaGAMYB-like overexpression Arabidopsis were obtained;

[0084] (10) The seeds of the T2 generation homozygous lines of TaGAMYB-like overexpression Arabidopsis were harvested as T3 generation (OE1 and OE2, respectively), and the next step of phenotype identification and analysis was carried out.

[0085] T0 generation represents the seeds and plants grown from the transformed generation; T1 generation represents the seeds and plants grown from the T0 generation; T2 generation represents the seeds and plants grown from the T1 generation; T3 generation represents the seeds and plants grown from the T2 generation; line represents the seeds or plant population produced by selfing of the same plant of the previous generation.

[0086] Example 4: PCR identification of transgenic Arabidopsis

[0087] The leaves of the two T2 generation homozygous lines of TaGAMYB-like overexpression Arabidopsis (OE1 and OE2) obtained in Example 3 were taken, the DNA in the leaves was extracted by CTAB method, and PCR amplification was carried out using upstream primer L1 and downstream primer R1.

[0088] The electrophoresis result of PCR product is shown in Figure 2 The PCR identification result shows that line OE1 and line OE2 are both TaGAMYB-like overexpression transgenic Arabidopsis lines.

[0089] Example 5: Real-time fluorescent quantitative PCR detection of transgenic Arabidopsis

[0090] The roots of the T3 generation homozygous transgenic lines (line OE1 and line OE2) and the roots of the wild type line of Columbia ecotype Arabidopsis (denoted as line WT) were photographed, and total RNA was extracted by Trizol method, and cDNA was obtained by reverse transcription with HiScript III RT SuperMix for qPCR (+gDNA wiper) reverse transcriptase after purification. Using wheat TaActin gene as an internal reference gene, real-time fluorescent quantitative PCR detection of TaGAMYB-like was carried out by qRT-PCR primers.

[0091] ABI QuantStudio3 real-time fluorescence quantitative PCR instrument, ChamQ SYBR Color qPCR Master Mix reagent, wheat TaActin gene expression level as internal reference, comparative threshold method for real-time fluorescence quantitative PCR result quantitative analysis, set fluorescence domain value, determine the cycle number Ct value under the fluorescence domain value. According to the Ct value, C=2 -△Ct , △Ct=Ct 目的基因 -Ct 内参基因 , calculate the average value of three repeated C values as the relative expression of the target gene

[0092] The reaction system is: 2×ChamQ SYBR Color qPCR Master Mix 5μL, 50×ROX Reference Dye II 0.2μL, upstream primer (10μM) 0.2μL, downstream primer (10μM) 0.2μL, 3-fold diluted first strand cDNA solution 0.5μL, ddH2O 3.9μL.

[0093] The qRT-PCR amplification procedure adopts three-step method:

[0094] (1) Pre-denaturation: 95℃ for 3min;

[0095] (2) Cycle reaction: 95℃ for 15s, 59℃ for 20s, a total of 40 cycles;

[0096] (3) Melting curve: 95℃ for 15s, 60℃ for 60s, 95℃ for 15s.

[0097] The primer sequence used in this step for real-time fluorescence quantitative PCR is as follows:

[0098] Actin-L: 5'-TATGCCAGCGGTCGAACAAC-3' (SEQ ID NO: 9);

[0099] Actin-R: 5'-GGAACAGCACCTCAGGGCAC-3' (SEQ ID NO: 10);

[0100] qRT-L: 5'-TGCCCACTGAAGTACGTAAG-3' (SEQ ID NO: 11);

[0101] qRT-R: 5'-ACCTACTCATTCAACCTGGT-3' (SEQ ID NO: 12).

[0102] qRT-PCR amplification results are shown inFigure 3 .Depend on Figure 3 It can be seen that the expression level of the exogenous gene TaGAMYB-like in strains OE1 and OE2 is significantly higher than that in strain WT, that is, the expression level of the exogenous gene TaGAMYB-like in transgenic Arabidopsis plants is significantly higher than that in wild-type plants.

[0103] Example 6: Phenotypic identification of transgenic Arabidopsis plants

[0104] 1. Phenotypic identification of germination rate during germination of TaGAMYB-like overexpression transgenic Arabidopsis lines

[0105] ABA stress treatment medium: Add 41.4g of MS solid powder to 1L of distilled water, stir thoroughly, adjust the pH to 6.0, sterilize at 121℃ for 15min, and when the medium cools to 40-50℃, add an appropriate amount of abscisic acid (ABA) to make the ABA concentration 0.5μM. Mix well and pour into round petri dishes to prepare ABA stress treatment medium with an ABA concentration of 0.5μM (0.5μM ABA group). The control group (CK group) did not add ABA.

[0106] Salt stress treatment medium: Add 41.4g MS solid powder and 11.7g NaCl to 1L distilled water, stir thoroughly, adjust the pH to 6.0, sterilize at 121℃ for 15min, and pour the medium into round petri dishes when it cools to 40-50℃ to prepare a salt stress treatment medium with a NaCl concentration of 200mM (200mM NaCl group). The control group (CK group) did not add NaCl.

[0107] The TaGAMYB-like overexpression transgenic Arabidopsis seeds (OE1, OE2) obtained in Example 3 and wild-type seeds (WT) of Colombian ecotype Arabidopsis were respectively spread on ABA stress treatment medium containing 0.5 μM ABA or salt stress treatment medium containing 200 mM NaCl and a control group (CK group) were set up. The germination of seeds in each group was observed and the germination rate was statistically analyzed after 10 days.

[0108] The germination rates of TaGAMYB-like overexpressing transgenic Arabidopsis seeds (OE1, OE2) and wild-type seeds (WT) of the Colombian ecotype Arabidopsis under ABA stress treatment are shown in the table below. Figure 4 .Depend on Figure 4It can be seen that under normal conditions (CK group), the germination rate of TaGAMYB-like overexpression transgenic Arabidopsis seeds OE1 and OE2 has no significant difference compared with the germination rate of wild type Arabidopsis seeds WT; under ABA stress (0.5 μM ABA group), the germination rate of TaGAMYB-like overexpression transgenic Arabidopsis seeds OE1 and OE2 is significantly reduced by 7.64% and 7.14% respectively compared with the germination rate of wild type Arabidopsis seeds WT.

[0109] The germination rate statistics results of TaGAMYB-like overexpression transgenic Arabidopsis seeds (OE1, OE2) and wild type seeds of Columbia ecotype Arabidopsis (WT) under salt stress treatment are shown in Table 2. Figure 5 Figure 5 It can be seen that under normal conditions (CK group), the germination rate of TaGAMYB-like overexpression transgenic Arabidopsis seeds OE1 and OE2 has no significant difference compared with the germination rate of wild type Arabidopsis seeds WT; under salt stress (200 mM NaCl group), the germination rate of TaGAMYB-like overexpression transgenic Arabidopsis seeds OE1 and OE2 is significantly increased by 32.26% and 29.03% respectively compared with the germination rate of wild type Arabidopsis seeds WT.

[0110] The above results show that the transgenic lines have higher sensitivity to ABA and higher tolerance to salt stress during seed germination.

[0111] 2, Phenotypic identification of TaGAMYB-like overexpression transgenic Arabidopsis lines at seedling stage

[0112] Square plate salt stress treatment medium: 41.4 g of MS solid dry powder and 8.775 g of NaCl are added to 1 L of distilled water, fully stirred and mixed, the pH value is adjusted to 6.0, 121 ℃ high temperature sterilization for 15 min, when the medium is cooled to 40-50 ℃, pour the plate (square plate), and the square plate salt stress treatment medium (NaCl group) with NaCl concentration of 150 mM is prepared. The control group (CK group) does not add NaCl.

[0113] The TaGAMYB-like overexpression transgenic Arabidopsis lines (OE1, OE2) and wild type lines of Columbia ecotype Arabidopsis (WT) are cultured in a incubator under normal conditions for 5 days, then transferred to square plate salt stress treatment medium added with NaCl for vertical culture (OE1-NaCl group, OE2-NaCl group, WT-NaCl group), and the control groups (OE1-CK group, OE2-CK group, WT-CK group) are set, and the root length of the plants is statistically analyzed after 10 days of culture.

[0114] The root growth of the control groups of each line is shown in Table 3.​Figure 6 , root length statistics results are shown in Figure 7 . It can be seen from Figure 6 and Figure 7 that under normal conditions, the root length of TaGAMYB-like overexpression transgenic Arabidopsis thaliana lines OE1 and OE2 has no significant difference compared with the root length of wild type Arabidopsis thaliana line WT.

[0115] The root growth of each line of square plate salt stress group is shown in Figure 8 , and the root length statistics results are shown in Figure 9 . It can be seen from Figure 8 and Figure 9 that under salt stress, the root length of TaGAMYB-like overexpression transgenic Arabidopsis thaliana lines OE1 and OE2 is significantly increased by 42.54% and 26.19% respectively compared with the root length of wild type Arabidopsis thaliana line WT.

[0116] 3. Phenotypic identification of TaGAMYB-like overexpression transgenic Arabidopsis thaliana lines in the adult stage

[0117] The TaGAMYB-like overexpression transgenic Arabidopsis thaliana lines (OE1, OE2) and the wild type strain of Columbia ecotype Arabidopsis thaliana (WT) were cultured in a round culture dish under normal conditions for 5d, and then were transferred to a plastic pot containing mixed culture medium (nutrient soil and vermiculite mixed in a mass ratio of 3:1) for culture, and were cultured under normal conditions for 12d, and were subjected to NaCl (150mM) stress treatment (NaCl group) and normal treatment (CK group), and the growth of plants in each group was observed on the 8th day, and the plant height was measured, and the fresh weight, dry weight, relative conductivity, chlorophyll content, and SOD and POD activities of plants in each group were determined on the 12th day.

[0118] The plant height statistics results of each group are shown in Figure 10 . It can be seen from Figure 10 that under normal conditions, the plant height of TaGAMYB-like overexpression transgenic Arabidopsis thaliana lines OE1 and OE2 has no significant difference compared with the plant height of wild type Arabidopsis thaliana line WT; under salt stress conditions, the plant height of TaGAMYB-like overexpression transgenic Arabidopsis thaliana lines OE1 and OE2 is significantly increased by 44.16% and 49.86% respectively compared with the plant height of wild type Arabidopsis thaliana line WT.

[0119] The fresh weight statistics results of plants in each group are shown in Figure 11 . It can be seen from Figure 11It can be seen that under normal conditions, the fresh weight of TaGAMYB-like overexpressing transgenic Arabidopsis thaliana lines OE1 and OE2 was not significantly different from that of wild-type Arabidopsis thaliana line WT; under salt stress conditions, the fresh weight of TaGAMYB-like overexpressing transgenic Arabidopsis thaliana lines OE1 and OE2 was significantly increased by 147.94% and 135.62% respectively compared with that of wild-type Arabidopsis thaliana line WT.

[0120] The dry weight statistics of each group of plants are shown in the figure. Figure 12 .Depend on Figure 12 It can be seen that under normal conditions, the dry weight of TaGAMYB-like overexpressing transgenic Arabidopsis thaliana lines OE1 and OE2 was not significantly different from that of wild-type Arabidopsis thaliana line WT; under salt stress conditions, the dry weight of TaGAMYB-like overexpressing transgenic Arabidopsis thaliana lines OE1 and OE2 was significantly increased by 73.90% and 60.87% respectively compared with that of wild-type Arabidopsis thaliana line WT.

[0121] The results of the measurements of plant height, fresh weight, and dry weight indicate that the TaGAMYB-like overexpression transgenic Arabidopsis thaliana lines have higher tolerance to salt stress.

[0122] The statistical results of the relative electrical conductivity of each group of plants are shown in the figure. Figure 13 .Depend on Figure 13 It can be seen that under normal conditions, the relative electrical conductivity of the TaGAMYB-like overexpressing transgenic Arabidopsis thaliana lines OE1 and OE2 was not significantly different from that of the wild-type Arabidopsis thaliana line WT; under salt stress, the relative electrical conductivity of the TaGAMYB-like overexpressing transgenic Arabidopsis thaliana lines OE1 and OE2 was significantly lower than that of the wild-type Arabidopsis thaliana line WT, decreasing by 16.92% and 13.85%, respectively.

[0123] When subjected to salt stress, cells are damaged, leading to electrolyte leakage and impairing vital functions, manifested as an increase in relative conductivity. The results showed that under salt stress, the TaGAMYB-like overexpressing transgenic Arabidopsis thaliana line suffered less cell damage and less electrolyte leakage than the wild-type Arabidopsis thaliana line WT, indicating that the former has stronger salt tolerance.

[0124] The statistical results of chlorophyll content of each group of plants are shown below. Figure 14 .Depend on Figure 14It can be seen that under normal conditions, the chlorophyll content of the TaGAMYB-like overexpression transgenic Arabidopsis lines OE1 and OE2 was not significantly different from that of the wild-type Arabidopsis line WT; however, under salt stress conditions, the chlorophyll content of the TaGAMYB-like overexpression transgenic Arabidopsis lines OE1 and OE2 was significantly increased by 15.74% and 14.74% respectively compared with the wild-type Arabidopsis line WT.

[0125] Chlorophyll is an important material basis for plant photosynthesis. Salt stress accelerates the decomposition of chlorophyll and leads to a decrease in net photosynthetic rate. The high chlorophyll content of the TaGAMYB-like overexpression transgenic Arabidopsis lines OE1 and OE2 indicates that their photosynthesis is less inhibited, allowing them to accumulate more photosynthetic assimilates and thus showing less impact from salt stress.

[0126] The results of SOD activity assays for each group of plants are shown below. Figure 15 .Depend on Figure 15 It can be seen that under normal conditions, the SOD activities of TaGAMYB-like overexpressing transgenic Arabidopsis thaliana lines OE1 and OE2 were not significantly different from those of wild-type Arabidopsis thaliana line WT; however, under salt stress conditions, the SOD activities of TaGAMYB-like overexpressing transgenic Arabidopsis thaliana lines OE1 and OE2 were significantly increased by 31.76% and 31.23% respectively compared with wild-type Arabidopsis thaliana line WT.

[0127] The results of POD activity assays for each group of plants are shown below. Figure 16 .Depend on Figure 16 It can be seen that under normal conditions, the POD activity of TaGAMYB-like overexpressing transgenic Arabidopsis thaliana lines OE1 and OE2 was not significantly different from that of wild-type Arabidopsis thaliana line WT; however, under salt stress conditions, the POD activity of TaGAMYB-like overexpressing transgenic Arabidopsis thaliana lines OE1 and OE2 was significantly increased by 15.77% and 11.62% respectively compared with wild-type Arabidopsis thaliana line WT.

[0128] SOD and POD are the main components of the enzymatic antioxidant system that scavenge reactive oxygen species (ROS). Under salt stress, the SOD and POD activities of the TaGAMYB-like overexpressing transgenic Arabidopsis lines OE1 and OE2 were significantly higher than those of the wild-type Arabidopsis line WT, indicating that OE1 and OE2 have a stronger ability to scavenge ROS and can effectively reduce membrane lipid peroxidation levels, thereby mitigating the harm of salt stress to the plants.

[0129] In summary, transforming the wheat gene TaGAMYB-like into the model plant Arabidopsis thaliana can significantly improve the salt tolerance of Arabidopsis thaliana.

[0130] It should be noted that the above examples are only used to clearly illustrate the present application, and are not intended to limit the embodiments of the present application. For those skilled in the art, on the basis of the above description, other different forms of changes or variations can also be made. Here, all the embodiments cannot be exhausted. Any obvious changes or variations derived from the technical scheme of the present application are still within the scope of protection of the present application.

Claims

1. The application of the wheat gene TaGAMYB-like in improving plant salt tolerance, characterized by: The nucleotide sequence of the wheat gene TaGAMYB-like is shown in SEQ ID NO:

1. Overexpression of this gene can improve the plant's tolerance to salt stress. The plant is wheat or Arabidopsis thaliana.

2. The application according to claim 1, characterized in that, The application includes the following steps: (1) PCR amplification and cloning of the wheat gene TaGAMYB-like; (2) The cloned wheat gene TaGAMYB-like was ligated into an expression vector to obtain an overexpression recombinant vector; (3) The overexpression recombinant vector was transformed into Agrobacterium to obtain the overexpression recombinant strain; (4) Transform the overexpressing recombinant strain into plants, screen and obtain wheat gene TaGAMYB-like overexpressing lines.

3. The application according to claim 2, characterized in that, The expression vector is super1300 containing the green fluorescent protein variant GFP-C.

4. The application according to claim 2, characterized in that, The Agrobacterium is Agrobacterium tumefaciens GV3101.

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