Use of hsfb2a in regulating oleic acid content in plant seeds

Abstract

The present application discloses HSFB2A Use in regulating the content of oleic acid in plant seeds. HSFB2A Use of the gene or protein or biological material related thereto in any one of the following: A1) increasing the content of oleic acid in Arabidopsis; A2) preparing a product for increasing the content of oleic acid in Arabidopsis; A3) cultivating Arabidopsis with increased content of oleic acid. The present application finds that the use of the gene for molecular breeding can direct the improvement of the fatty acid composition of Arabidopsis, specifically significantly increasing the content of oleic acid, which provides a new regulation target and gene tool for solving the core bottleneck of current high-oleic-acid breeding, and has important agricultural application value. HSFB2A The present application discloses

Description

Technical Field

[0001] This invention belongs to the field of biology, specifically relating to HSFB2A Application in regulating oleic acid content in plant seeds. Background Technology

[0002] Oleic acid (C18:1), as a monounsaturated fatty acid, is a core indicator for evaluating the quality of vegetable oils. High-oleic vegetable oils with an oleic acid content ≥75% are widely recognized as healthy and high-quality oils, and are a core focus of global oilseed crop breeding. On the one hand, high-oleic oils have received widespread attention for their nutritional and health benefits: increasing the oleic acid ratio can improve the stability of oils (antioxidant, extended shelf life) and has many beneficial effects on human cardiovascular health; high-oleic vegetable oils can lower LDL cholesterol and raise HDL cholesterol, reducing the risk of cardiovascular disease; they have strong oxidative stability, a high smoke point, and are less likely to produce harmful substances during high-temperature cooking; they also have antioxidant, anti-inflammatory, and fat-soluble vitamin absorption-promoting effects, making them an important direction for optimizing the quality of vegetable oils. On the other hand, from an industrial perspective, high-oleic vegetable oils have a long shelf life, good processing tolerance, and increase economic added value, thus becoming a priority target for breeding improvement.

[0003] Current mainstream strategies for breeding high-oleic acid varieties focus on the targeted modification of key fatty acid synthesis enzyme genes, with FAD2 (fatty acid desaturase 2) and FAE1 (fatty acid elongation enzyme 1) as the core targets. By knocking out or silencing FAD2, the conversion of oleic acid to linoleic acid is blocked; by knocking out or silencing FAE1, the elongation of oleic acid into long-chain fatty acids is inhibited, thereby significantly increasing oleic acid content. FAD2 / FAE1 mutants created using CRISPR / Cas9, RNAi, and other technologies have been successfully applied in crops such as rapeseed, soybean, and peanut, and some varieties have been commercialized, laying an important foundation for high-oleic acid breeding.

[0004] However, the existing "enzyme gene blocking" strategy has technical bottlenecks that restrict the breakthrough development of high oleic acid breeding: (1) Narrow genetic basis and single gene resources: Over-reliance on a few enzyme structural genes such as FAD2 / FAE1, single regulatory pathway, easy to lead to variety homogenization, and difficulty in breaking through the existing upper limit of oleic acid content (most crops are stable at 75%-85%). (2) Prominent contradiction in trait synergy: High oleic acid trait is often linked with undesirable agronomic traits such as low oil content, low yield, and weakened stress resistance, making it difficult to achieve synergistic improvement of "high oleic acid, high yield, and high oil content". (3) Limited regulatory mechanism: Enzyme gene modification is only "blocking" regulation, which only inhibits the downstream metabolism of oleic acid and cannot activate the oleic acid synthesis pathway from the transcriptional regulation level, lacking the ability to systematically regulate the fatty acid metabolism network.

[0005] Transcription factors, acting as "molecular switches" of gene expression, can systematically regulate the expression of key genes involved in multiple stages of fatty acid synthesis, desaturation, and accumulation by binding to target gene promoters. They represent ideal targets for overcoming current limitations. Therefore, developing novel transcription factor tools capable of bidirectional and precise regulation of the fatty acid metabolism network has significant theoretical and practical value. Summary of the Invention

[0006] To address the problems in existing technologies, this invention is the first to discover a transcription factor. HSFB2A This gene has not been reported to participate in fatty acid metabolism regulation, but its specific overexpression in Arabidopsis seeds significantly increases oleic acid content. Unlike existing enzyme genes such as FAD2 / FAE1, this gene... HSFB2A Transcription factors can positively activate the oleic acid synthesis pathway at the transcriptional level, breaking through the limitations of traditional "blocking" regulation and providing new regulatory targets and gene tools for solving the core bottleneck of current high oleic acid breeding.

[0007] First aspect of protection of the present invention HSFB2A The application of genes or proteins or related biological materials in any of the following: A1) Increase the oleic acid content in plants; A2) Prepare products that increase the oleic acid content in plants; A3) Cultivate plants with increased oleic acid content.

[0008] In some embodiments, the HSFB2A Nucleotide sequence of the gene coding region (BnaC03G0559700ZS).

[0009] ATGGTGAGCGATAACGGTGACCCGATGATGGTAATAACAAATGAGTCTCAGAGAACTGTTCCGACGCCGTTTCTCATCAAGACCTTTAACCTCGTTGAAGATCCTTCCATCGACGACGTCATCTCATGGAACGAAGACGGTTCCTCCTTCGTCGTCTGGAACCCGACTGACTTCGCTAGAGACTTGCTTCCTAAACACTTTAAACACAACAACTTCTCCAGCTTTGTCCGTCAGCTCAACACTTACGGGTTCAAGAAAGTAGTCCCTGATCGGTGGGAGTTTTCGAATGATTTCTTCAGGAGAGGAGAGAAACGTCTTCTCAAAGAGATCCAACGTCGAAAGTTAACGCCTCAAGCAGCTGTTACTCCGCCGCCTGAAGTGGTTGCGGCGGCGGCGGCTCAGAGAGTTCAGACGACGAAAACGGTCGTGTCTCCGTCGAGTTCTGCGGAAGATCAGGCGGTATCTCCGTCGTGGTATTGTCAAACCGGAAACGGTGGTGGCTTGTCTGTTGAGCTGTTAGAAGAGAACGAGAAGCTTCGGAGTCAGAACATCCAGCTTAACCGTGAGCTTACTCAGATGAAATCCCTCTGCGATAACATCTTCAGCCTCATGTCCAACTTCTCCGGATCTCCTCCTGAACGGAGCTCATCTCCCGGAGCTAGTACCAGTCATGACGCGATGAAAACGGTGGAGTTTTTACCGGCGAAGCGGTTTTCGGTTGAGACGGAGGGTGAGGAAGAAGCGAGTCATAGGCTTTTCGGTGTTTCGATAGGGCTGAAACGGACGAGAAGCGAAGGTGTTCAGGTGAAGGCGGTGGGAGAGAGACCCGGCGGAGGAGGGGAGAAGGAGGAGACGCCGTGGCTGAGACACTATAATAGGACCAATCAGAGAGTCTGTAATTGA(SEQ ID No.1) In some embodiments, the HSFB2A amino acid sequence of the protein comprises the sequence set forth in SEQ ID No.2.

[0010] MVSDNGDPMMVITNESQRTVPTPFLIKTFNLVEDPSIDDVISWNEDGSSFVVWNPTDFARDLLPKHFKHNNFSSFVRQLNTYGFKKVVPDRWEFSNDFFRRGEKRLLKEIQRRKLTPQAAVTPPPEVVAAAAAQRVQTTKTVVSPSSSAEDQ AVSPSWYCQTGNGGGLSVELLEENEKLRSQNIQLNRELTQMKSLCDNIFSLMSNFSGSPERSSSPGASTSHDAMKTVEFLPAKRFSVETEGEEEASHRLFGVSIGLKRTRSEGVQVKAVGERPGGGGEKEETPWLRHYNRTNQRVCN(SEQ ID No.2) In some implementations, overexpression HSFB2A Genes or proteins can increase the oleic acid content in plants.

[0011] In some embodiments, the biomaterial is at least one of the following: B1) includes the above HSFB2A Nucleic acid constructs of genes or expression HSFB2A Nucleic acid constructs of proteins; B2) Genetically engineered bacteria containing the nucleic acid constructs described in B1).

[0012] In some embodiments, the starting plasmid of the nucleic acid construct is selected from pBinGlyRed3.

[0013] In some embodiments, the plant is selected from one or both of rapeseed and Arabidopsis thaliana.

[0014] A second aspect of the present invention protects a method for cultivating plants with increased oleic acid content, comprising: increasing the oleic acid content in recipient plants. HSFB2A The protein content and / or activity were improved to obtain plants with increased oleic acid content.

[0015] In some embodiments, the method for increasing the content and / or activity of HSFB2A protein in the recipient plant is to overexpress it in the recipient plant. HSFB2A protein.

[0016] In some embodiments, the overexpression method is to... HSFB2A The protein-coding gene was introduced into the recipient plant.

[0017] In some embodiments, the recipient plant is selected from one or both of rapeseed and Arabidopsis thaliana.

[0018] A third aspect of the present invention protects a method for increasing the oleic acid content in plants, comprising: increasing the oleic acid content in recipient plants. HSFB2A Protein content and / or activity.

[0019] In some embodiments, the enhancement of recipient plants HSFB2A The method for determining the protein content and / or activity is to overexpress the HSFB2A protein in the recipient plant.

[0020] In some embodiments, the overexpression method is to... HSFB2A The protein-coding gene was introduced into the recipient plant.

[0021] In some embodiments, the recipient plant is selected from one or both of rapeseed and Arabidopsis thaliana.

[0022] Compared with the prior art, the present invention has the following beneficial effects: 1) This invention is the first to clearly define HSFB2A Biological functions of genes in regulating oleic acid in plants and their application value.

[0023] 2) This invention discovers and utilizes HSFB2A Molecular breeding using genes can directionally improve the fatty acid composition of plants, specifically by significantly increasing oleic acid content. This provides a novel regulatory target and gene tool for addressing the core bottleneck of high oleic acid breeding, and has significant agricultural application value.

[0024] 3) This invention establishes a highly efficient overexpression method. HSFB2A Gene technology systems can achieve [the following]: HSFB2A Precise regulation of gene expression levels provides a practical molecular tool for targeted improvement of oil quality in oilseed crops such as rapeseed. Attached Figure Description

[0025] Figure 1 The rapeseed source for the present invention BnaHSFB2A The results of oleic acid percentage content in seeds after overexpression in Arabidopsis thaliana are shown in the figure. CK represents the wild-type control in Arabidopsis thaliana; BnaHSFB2A-OE#1-3 represents the overexpression lines of BnaHSFB2A in Brassica napus, and three stars indicate highly significant differences (P<0.001, n>4).

[0026] Figure 2 This is a spectrum of pBinGlyRed3 of the present invention.

[0027] Figure 3 The map shows the overexpression vector constructed in Example 2 of this invention. Detailed Implementation

[0028] The present invention will be further described below with reference to the accompanying drawings and specific embodiments, but the scope of protection of the present invention is not limited to the described contents. Unless otherwise specified, the methods in this embodiment are operated according to conventional methods, and the reagents used are conventional reagents unless otherwise specified.

[0029] Plant material: The developing seeds of Zhongshuang 11 Brassica napus were selected and purchased from Wuhan Zhongyou Seed Industry Technology Co., Ltd.

[0030] Strains and vectors: Agrobacterium ( Agrobacterium tumefaciens (Strain GV3101 was purchased from Shanghai Weidi Biotechnology, CAT#: AE1001); the overexpression vector pBinGlyRed3 was kindly provided by Professor Zhang Chunyu of Huazhong Agricultural University (plasmid map can be found here). Figure 2 ).

[0031] Example 1: Brassica napus ZS11 BnaHSFB2A Cloning of genes 1.1. Seed RNA extraction during the development of Brassica napus ZS11 All pipette tips, centrifuge tubes, and solutions used in the following operations were free of RNase contamination, and gloves were worn during all operations.

[0032] 1) Preparation of extraction solution: Take 500 μL of RL lysis buffer (Beijing Tiangen, catalog number DP441), then add 5 μL of β-mercaptoethanol and mix well.

[0033] 2) Homogenization: Remove the pods from the developing seeds in liquid nitrogen, quickly grind them into powder, take 0.1 g, add 500 μL of RL lysis buffer, and vortex vigorously to mix. Centrifuge at 12000 rpm for 5 min, and collect the supernatant.

[0034] 3) Transfer all the supernatant from step 2 to the CS filter column, centrifuge at 12000 rpm for 2 min, and carefully aspirate the supernatant from the collection tube into a new RNase-free centrifuge tube, avoiding contact between the pipette tip and cell debris in the collection tube as much as possible.

[0035] 4) Add 0.5 times the volume of supernatant to the centrifuge tube and mix thoroughly. Then transfer it to the adsorption column CR3, centrifuge at 12000 rpm for 1 min, discard the waste liquid in the collection tube, and put the adsorption column CR3 back into the collection tube.

[0036] 5) Add 350 μL of protein removal solution RW1 to the adsorption column CR3, centrifuge at 12000 rpm for 1 min, discard the waste liquid in the collection tube, and put the adsorption column CR3 back into the collection tube.

[0037] 6) Add 80 μL of DNase I digestion solution to the adsorption column CR3 and incubate at room temperature for 15 min to digest DNA.

[0038] 7) Add 350 μL of protein removal solution RW1 to the adsorption column CR3, centrifuge at 12000 rpm for 1 min, discard the waste liquid in the collection tube, and put the adsorption column CR3 back into the collection tube.

[0039] 8) Add 500 μL of washing buffer RW to the adsorption column CR3, let stand at room temperature for 2 min, centrifuge at 12000 rpm for 1 min, discard the waste liquid in the collection tube, and put the adsorption column CR3 back into the collection tube. Repeat step 8) once.

[0040] 9) Centrifuge at 12000 rpm for 2 min and discard the waste liquid. Place the adsorption column CR3 at room temperature for several minutes to thoroughly dry any residual rinsing liquid in the adsorption material.

[0041] 10) Place the adsorption column CR3 into a new RNase-Free centrifuge tube, add 50 μL of RNase-Free water dropwise to the center of the adsorption membrane, incubate at room temperature for 2 min, and centrifuge at 12000 rpm for 2 min to obtain the RNA solution. Measure the RNA concentration and the OD ratio (260:280). If the ratio is between 1.8 and 2.0, the RNA is considered usable and can be stored at -70℃ for later use, but should not be stored for more than one week.

[0042] 1.2 Synthesis of cDNA strands from reverse transcription of total RNA The ReverTra Ace-α-reverse transcription kit from Toyobo Corporation was used.

[0043] 1) Thermal denaturation of RNA: Add 10 μL of RNA (1 μg / μL), 2 μL of Oligo(dT) (10 pmol / μL), and 12 μL of DEPC water sequentially to an ice box. Mix gently, centrifuge in a hand centrifuge to remove residual liquid from the tube wall, and react at 65°C for 5 min in a metal bath and immediately place on ice. This step is optional and is used to disrupt the tertiary or secondary structure of RNA, causing the strand to open.

[0044] 2) Preparation of reaction solution: Add 8 μL of 5×Reaction Buffer, 2 μL of RNase Inhibitor (10 U / μL), 4 μL of dNTPs Mixture (10 mM), and 2 μL of ReverTra Ace-α-reaction solution to a 40 μL reaction mixture on ice. Gently mix and incubate in a metal bath at 42℃ for 1 h, followed by heat denaturation at 85℃ for 10 min to terminate the reaction. After the reaction is complete, store the reaction product at -20℃ for later use.

[0045] 1.3 PCR amplification and sequencing of the full-length gene and coding region Primer design reference sequences were primarily obtained from the Brassica napus ZS11 genome database. Specific primer pairs were designed based on the predicted sequence information from the genome database. BnaHSFB2A The gene coding region sequence was amplified.

[0046] Add 2 μL of 10×PCR Buffer, 2 μL of 25 mM MgSO4, and 2 μL of dNTP (2.5 mM) sequentially to ice. BnaHSFB2A Upstream primer (25 μM) 0.3 μL, BnaHSFB2 Downstream primer (25 μM) 0.3 μL, cDNA (200 ng / μL) 1 μL, KOD plus (1 U / μL) 1 μL, ddH2O 11.8 μL, total volume 20 μL.

[0047] BnaHSFB2A Upstream primer sequence: 5'-ATGGTGAGCGATAACGGTGACCC-3' (SEQ ID No. 3); BnaHSFB2A Downstream primer sequence: 5'-TCAATTACAGACTCTCTGATTGGTCCTA-3' (SEQ ID No. 4).

[0048] The reaction procedure is as follows: Pre-denaturation at 94℃ for 3 min, denaturation at 94℃ for 30 s, annealing at 60℃ for 30 s, extension at 68℃ for 40 s-1 min, and extension at 68℃ for another 10 min, for a total of 30 cycles.

[0049] After amplification, the amplification products were subjected to agarose gel electrophoresis. The target band was then observed under UV light. The target band was cut off with a blade and transferred to a centrifuge tube. The DNA fragment was recovered using an agarose gel extraction kit and ligated into the T vector. On ice, 5 μL of Solution I, 4 μL of the PCR product, and 1 μL of pMD18-T vector (50 ng / μL) were added to a PCR tube, bringing the total volume to 10 μL. The tube was then placed in a metal bath and incubated overnight at 16°C. The ligation product was added to competent E. coli cells and mixed. The tube was incubated on ice for 30 min, then heat-shocked in a 42°C water bath for 90 s, and immediately placed on ice for 3 min. 700 μL of LB medium was added, and the tube was incubated at 37°C and 200 rpm for 30 min. 200 μL of the culture was transferred to solid LB medium (100 µg / mL) containing ampicillin and spread evenly with a spreader.

[0050] The plates were inverted and incubated overnight at 37°C. Positive plaques were sent to the company for sequencing. The sequence fragments obtained contained coding regions and were named as follows: BnaHSFB2A Its amino acid and nucleotide sequences are shown in SEQ ID No. 2 and SEQ ID No. 1, respectively.

[0051] Example 2: Construction of overexpression vector, transformation into Arabidopsis thaliana, and fatty acid analysis in transgenic lines. 2.1 Construction of plant expression vectors and Agrobacterium-mediated transformation 1) Cloned using Example 1 BnaHSFB2A The fragment was used as a template and amplified using the high-fidelity enzyme KOD plus from TOYOBO. The amplified product was recovered via agarose gel for later use, which is the amplified fragment.

[0052] 2) Take 40 μL of plant expression vector pBinGlyRed3 plasmid, add 4 μL EcoR I enzyme and 5 μL enzyme digestion buffer, mix well, and digest at 37℃ for 30 min. The vector is then recovered via agarose gel for later use, which is the digested plasmid.

[0053] 3) Take 2 μL of the amplified fragment obtained in step 1) and 2 μL of the enzyme-digested plasmid obtained in step 2), add 1 μL of infusion kit mix, mix well, and ligate at 50℃ for 20 min to obtain the ligation product.

[0054] The ligation product was incubated in a water bath at 42°C for 90 seconds, followed by ice shock for 2 minutes to transform *E. coli* DH5α, resulting in positive plaques. The plasmid was then extracted for later use; this is the recombinant plasmid (i.e., the overexpression vector; plasmid map can be found in...). Figure 3 ).

[0055] The recombinant plasmid carries the soybean globulin promoter (originally present in the pBinGlyRed3 plasmid). BnaHSFB2A The coding region sequence of the gene is specifically expressed in seeds. The recombinant plasmid was transformed into Agrobacterium GV3101, and positive Agrobacterium strains were screened: 200 ng of the recombinant plasmid was added to a 2 mm electroporation cuvette (Bio-Rad Laboratories, USA) containing GV3101 competent cells (purchased from Shanghai Weidi Biotechnology, CAT#: AE1001). After electroporation at 2.5 KV for 5 ms, 1 ml of LB liquid medium was added, and the culture was allowed to stand for 30 min. Then, 50 μL of the bacterial culture was spread onto an LB agar plate containing 50 μg / ml kanamycin. Two days later, single Agrobacterium-positive plaques were selected and shaken to activate the bacteria.

[0056] 2.2 Agrobacterium infection and transformation of Arabidopsis thaliana 1) Add the positive strain obtained in step 2.1 to LB liquid medium (0.5% yeast extract, 1% peptone, 1% sodium chloride) containing gentamicin (50 μg / mL), kanamycin (50 μg / mL), and rifampin (25-50 μg / mL), and incubate overnight at 28°C and 200 rpm.

[0057] 2) Then, centrifuge at 5000 rpm for 3 min to collect the bacterial cells, and dilute Agrobacterium with transformation osmotic buffer (5% sucrose, 0.02% Silwet L-77 (Beijing Lanbolide, catalog number: S9430)) to an OD600 value of about 0.8.

[0058] 3) Select Arabidopsis thaliana in the early flowering stage for flower-dipping conversion. Before conversion, the siliques of Arabidopsis thaliana should be cut off. Immerse the Arabidopsis thaliana inflorescence in the above-mentioned conversion solution with an OD600 value of 0.8 for 0.5-1 min.

[0059] 4) Place the Arabidopsis seedlings in dark, humid conditions overnight. Then return the Arabidopsis to the culture room and repeat the transformation after 3-5 days. Harvest the seeds after the Arabidopsis matures.

[0060] Because the plant expression vector carries the DsRed3 fluorescent protein, when the seeds are excited to release fluorescence under 520 nm green light, the red seeds that pass through the filter are the transgenic T1 generation.

[0061] 2.3 Preliminary screening of transgenic T1 generation single-copy lines After the T1 generation plants matured, their seeds were harvested, which became the T2 generation. Copy number screening was then performed on these seeds. Transgenic T2 seeds were excited under 520nm green light using a fluorescence stereomicroscope (Zeiss, Axio Zoom. Vl6), and bright-field and excitation-light-based images were captured after passing through a red filter.

[0062] The collected photos were used to count the number of seeds using statistical software such as Wanshen CG Seeding Software or ImageJ, and the final ratio of red seeds to non-red seeds was obtained.

[0063] Select strains with a calculated ratio close to 3:1, and choose 10 red seeds from them for sowing. After the seeds mature, harvest the seeds as T3 generation seeds.

[0064] The T3 generation is divided into three groups, namely BnaHSFB2A- OE#1、 BnaHSFB2A- OE#2 BnaHSFB2A- OE#3.

[0065] Meanwhile, wild-type Arabidopsis thaliana was used as the wild control group (CK).

[0066] 2.4 Fatty acid analysis and expression analysis of overexpression lines 2.4.1 Extraction and Methyl Esterification of Oils 1) Weigh 5-10 mg of Arabidopsis thaliana T3 generation seeds and put them into a 10 mL glass test tube.

[0067] 2) Add 1.5 mL of 2.5% H₂SO₄ methanol solution (containing 0.01% BHT), 200 μL of C17:0 toluene solution (concentration 2 mg / mL), and 0.4 mL of toluene solution sequentially. After filling with nitrogen gas, be sure to tighten the cap to prevent evaporation at high temperatures. C17:0 (where 0 represents the number of double bonds) represents the internal reference fatty acid (Sigma-Aldrich, H3500), heptadecanoic acid, with the chemical formula C... 17 H 34 O2 has a molecular weight of 270.45 g / mol.

[0068] 3) Heat in a 99℃ water bath for 1 h, remove and let cool, then add 1.8 mL of double-distilled water and 1 mL of n-hexane in sequence.

[0069] 4) After mixing, let stand for 12 h, use a 1 mL syringe to draw up the supernatant, and then filter it through a 0.45 μm pore size into a sample vial for testing.

[0070] 2.4.2 Gas Chromatographic Analysis of Fatty Acid Methyl Esters The gas chromatograph used was an Agilent 7890A, and the column was an Agilent J&W GC Columns. The injection volume was 1 μL, and the column oven heating program was: initial temperature 180℃, heating to 225℃ at a rate of 10℃ / min, and holding for 7 min. The constant flow rate of the column was 1 mL / min, and the carrier gas was N2.

[0071] Peak area was calculated using software provided by Agilent Chemical Workstation. The internal standard method was used to calculate oil content (C17:0 was used as the internal standard for determining total fatty acids). The calculation formula is: m(C17:0 mass) / s(C17:0 peak area) = m(oil mass) / s(peak area of ​​fatty acids other than C17:0).

[0072] BnaHSFB2A Results of fatty acid composition of transgenic lines are shown below Figure 1 .

[0073] from Figure 1 It can be known that Arabidopsis pilosula BnaHSFB2A Oleic acid (C) in the dry seeds of seed-overexpressing strains 18 H 34O2 (i.e., C18:1), where 1 indicates that the percentage of double bonds is 25-27%, which is 10% to 13% higher than that of wild type (14-15%).

[0074] This invention is not limited to the specific textual description above. Various changes can be made to this invention within the scope outlined in the claims, and all such changes are within the scope of this invention.

Claims

1. Application of the HSFB2A gene or HSFB2A protein or related biological materials in any of the following: A1) Increase the oleic acid content in Arabidopsis thaliana; A2) Prepare products that increase the oleic acid content in Arabidopsis thaliana; A3) Cultivate Arabidopsis thaliana with increased oleic acid content; The nucleotide sequence of the HSFB2A gene is shown in SEQ ID No.

1. Overexpression of the HSFB2A gene can increase the oleic acid content in Arabidopsis thaliana. The biomaterial is at least one of the following: B1) A nucleic acid construct containing the HSFB2A gene or a nucleic acid construct expressing the HSFB2A protein; B2) Genetically engineered bacteria containing the nucleic acid constructs described in B1).

2. The application as described in claim 1, characterized in that, The amino acid sequence of the HSFB2A protein is shown in SEQ ID No.

2.

3. The application as described in claim 1, characterized in that, The starting plasmid of the nucleic acid construct was selected from pBinGlyRed3.

4. A method for cultivating Arabidopsis thaliana with increased oleic acid content, characterized in that, include: To increase the content and / or activity of HSFB2A protein in Arabidopsis thaliana, Arabidopsis thaliana with increased oleic acid content was obtained. The nucleotide sequence of the gene encoding the HSFB2A protein is shown in SEQ ID NO:

1.

5. A method for increasing the oleic acid content in Arabidopsis thaliana, characterized in that, include: To increase the content and / or activity of HSFB2A protein in Arabidopsis thaliana, wherein the nucleotide sequence of the gene encoding the HSFB2A protein is shown in SEQ ID NO:

1.

6. The method as described in claim 4 or 5, characterized in that, The method for increasing the content and / or activity of HSFB2A protein in Arabidopsis thaliana is to overexpress HSFB2A protein in Arabidopsis thaliana.

7. The method according to claim 6, characterized in that, The overexpression method involves introducing the gene encoding the HSFB2A protein into Arabidopsis thaliana.

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