Application of CsABF8 gene in regulation and control of plant drought resistance

Abstract

The invention provides application of a CsABF8 gene in regulation and control of plant drought resistance, and belongs to the technical field of molecular breeding. According to the application disclosed by the invention, a drought-sensitive variety 'Fuyun No.6' ('FY') and a drought-tolerant variety 'Taigu No.12' ('TC') are taken as experimental materials, and the CsABF8 gene is determined to be a potential drought-resistant target gene through drought stress treatment and non-targeted metabolome screening of differential metabolites. The CsABF8 gene can respond to drought stress, and the content of raffinose is increased by regulating CsGolS1, CsGolS2 and CsRaf6 promoters in a tea tree body, so that the tea tree can cope with the drought stress.

Description

Technical Field

[0001] The invention belongs to the technical field of molecular breeding, and in particular relates to the application of CsABF8 gene in regulating plant drought resistance. Background Art

[0002] Tea tree is an important perennial evergreen leaf cash crop. Tea yield is directly affected by water supply. Water shortage can lead to a 33% to 40% reduction in tea yield. The plant hormone Abscisic acid (ABA) plays a key role in integrating multiple abiotic stress signals such as drought, cold and salinity and inducing protective stress responses. When plants are subjected to drought stress, ABA accumulates rapidly. ABA can activate transcriptional and post-transcriptional gene expression, regulate stomatal closure and metabolite changes, and thus resist drought. Therefore, discovering genes related to the regulation of tea tree drought response and clarifying their regulatory mechanisms are of great significance for cultivating drought-resistant tea varieties that efficiently utilize water resources.

[0003] However, there is no report on the involvement of CsABFs genes in drought stress in tea plants. Summary of the invention

[0004] The purpose of the present invention is to provide an application of the CsABF8 gene in regulating plant drought resistance. The CsABF8 gene can be used to regulate the drought resistance of tea plants.

[0005] The present invention provides application of CsABF8 gene in regulating plant drought resistance; the amino acid sequence of the protein encoded by the CsABF8 gene is shown in SEQ ID NO.1.

[0006] Preferably, the nucleotide sequence of the CsABF8 gene is as shown in SEQ ID NO.2.

[0007] Preferably, the application includes the application of negatively regulating the CsABF8 gene in improving plant drought resistance or the application of positively regulating the CsABF8 gene in constructing a drought-sensitive plant model.

[0008] Preferably, the negative regulation includes transient silencing; the reagent used for the negative regulation includes an antisense oligonucleotide combination targeting the CsABF8 gene; the antisense oligonucleotide combination includes the first antisense oligonucleotide shown in SEQ ID NO.8, the second antisense oligonucleotide shown in SEQ ID NO.9 and the third antisense oligonucleotide shown in SEQ ID NO.10.

[0009] Preferably, the positive regulation includes transient overexpression; the reagent used for the positive regulation includes a CsABF8 gene overexpression vector plasmid; the CsABF8 gene overexpression vector plasmid uses pCAMBIA 1305 as a backbone plasmid, into which the CsABF8 gene is inserted.

[0010] Preferably, the plant comprises tea tree.

[0011] The present invention also provides a method for cultivating drought-resistant plants, comprising the following steps: injecting an antisense oligonucleotide combination solution targeting the CsABF8 gene into mature leaves of tea trees to achieve instantaneous silencing of the CsABF8 gene in the plant.

[0012] The present invention also provides a method for constructing a drought-sensitive plant model, comprising the following steps: introducing a CsABF8 gene overexpression vector plasmid into Agrobacterium tumefaciens to obtain a recombinant bacterium; and infecting a plant with the recombinant bacterium to obtain a drought-sensitive plant model.

[0013] The present invention also provides the use of the CsABF8 gene in regulating the raffinose content of plants; the amino acid sequence of the protein encoded by the CsABF8 gene is shown in SEQ ID NO.1.

[0014] The present invention also provides the use of CsABF8 gene in regulating CsGolS1 / 2 promoter and / or CsRaf6 promoter; the amino acid sequence of the protein encoded by the CsABF8 gene is shown in SEQ ID NO.1.

[0015] The present invention provides an application of the CsABF8 gene in regulating plant drought resistance; the amino acid sequence of the protein encoded by the CsABF8 gene is shown in SEQ ID NO.1. The present invention uses the drought-sensitive variety 'Fuyun No. 6' ('FY')' and the drought-tolerant variety 'Taicha No. 12' ('TC') as experimental materials, and through drought stress treatment and non-targeted metabolomics screening of differential metabolites, it is clarified that the CsABF8 gene is a potential drought-resistant target gene. The CsABF8 gene can respond to drought stress, and in the tea tree, by regulating the CsGolS1, CsGolS2 and CsRaf6 promoters to increase the raffinose content, thereby enabling the tea tree to cope with drought stress. BRIEF DESCRIPTION OF THE DRAWINGS

[0016] In order to more clearly illustrate the embodiments of the present invention or the technical solutions in the prior art, the drawings required for use in the embodiments will be briefly introduced below. Obviously, the drawings described below are only some embodiments of the present invention. For ordinary technicians in this field, other drawings can be obtained based on these drawings without paying creative labor.

[0017] Figure 1 This is a schematic diagram of the CsABF8 gene silencing experiment treatment;

[0018] Figure 2 This is the result of the determination of the expression level of related genes after inhibition treatment and the determination of the raffinose content in the leaves;

[0019] Figure 3 Schematic diagram of the potential binding region between CsABF8 and the promoters of CsGolS1, CsGolS2, and CsRaf6;

[0020] Figure 4 Schematic diagram of overexpression of pCAMBIA 1305-CsABF8;

[0021] Figure 5 The figure shows the expression changes of galactose pathway related genes and the determination results of raffinose content in leaves after overexpression of pCAMBIA 1305-CsABF8;

[0022] Figure 2 and Figure 5 All data were obtained from at least three independent biological replicates (n ≥ 3), *P < 0.05; **P < 0.01; ***P < 0.001, and asterisks represent significant differences under independent sample T-test. DETAILED DESCRIPTION

[0023] The present invention provides an application of the CsABF8 gene in regulating plant drought resistance; the amino acid sequence of the protein encoded by the CsABF8 gene is shown in SEQ ID NO.1, specifically:

[0024] MNFKNFGNEPPEEGGAGGGGRPLGIFPLARQTSIYSLTFDEFQSTIGGSGKDFGSMNMDELLKNIWRTEETQTVGTTSGLQGQGQGQEGVAPNGHLQRQGSLTLPRTLSQKTVDE VWRDLAKEYGGGKDGIGAVNSNQPQRQPTLGEMTLEEFLVRAGVVREELQLPHGNPNNTGFGDLSRPNNNTALGFAFQQMGQNTGLMGNRIPESNNQNTIQSANLPLNVNGVRST QQQQQQQQQQRGQQQQQLFPKQPNMGYANPMSIPSSTQMSSPGFRGGIVGIADSTITNNLVQSAALQGGGMGMMGLGAGAVATGSPAFSSDGLGKSNGDTSSVSPVPYVFNGGL RGRKSSHAVEKVVERRQRRMIKNRESAARSRARKQAYTMELEAEVAKLKEEENQELRKKQMNITIINFVAGGNNGIAEKSGLGDDEHAAGRKETMPEANTDWAMVKGRDYALIIQI.

[0025] In the specific implementation of the present invention, the nucleotide sequence of the CsABF8 gene is shown in SEQ ID NO.2, specifically:

[0026]

[0027] In the specific implementation process of the present invention, the application includes the application of negatively regulating the CsABF8 gene in improving plant drought resistance or the application of positively regulating the CsABF8 gene in constructing a drought-sensitive plant model.

[0028] In the specific implementation process of the present invention, the negative regulation includes transient silencing; the reagent used for the negative regulation includes an antisense oligonucleotide combination targeting the CsABF8 gene; the antisense oligonucleotide combination includes the first antisense oligonucleotide shown in SEQ ID NO.8, the second antisense oligonucleotide shown in SEQ ID NO.9 and the third antisense oligonucleotide shown in SEQ ID NO.10.

[0029] In one embodiment of the present invention, the antisense oligonucleotide combination consists of a first antisense oligonucleotide shown in SEQ ID NO.8, a second antisense oligonucleotide shown in SEQ ID NO.9 and a third antisense oligonucleotide shown in SEQ ID NO.10; the first antisense oligonucleotide, the second antisense oligonucleotide and the third antisense oligonucleotide simultaneously act on the upstream, midstream and downstream of the CsABF8 gene, thereby inhibiting the expression of the CsABF8 gene.

[0030] In the specific implementation process of the present invention, the positive regulation includes transient overexpression; the reagent used in the positive regulation includes CsABF8 gene overexpression vector plasmid; the CsABF8 gene overexpression vector plasmid uses pCAMBIA 1305 as a backbone plasmid, and is inserted with the CsABF8 gene.

[0031] In the specific implementation of the present invention, the plant includes tea trees. In the specific implementation of the present invention, the varieties of the tea trees include 'Taicha No. 12' or 'Shucha Zao'; the tea trees include tea seedlings; and the tea seedlings include tea tree cutting seedlings.

[0032] The present invention also provides a method for cultivating drought-resistant plants, comprising the following steps:

[0033] The antisense oligonucleotide combination solution targeting the CsABF8 gene was injected into the mature leaves of the tea plant to achieve instantaneous silencing of the CsABF8 gene in the plant.

[0034] In the specific implementation process of the present invention, the plant includes tea tree; the plant leaves include the second and third leaves of tea seedlings; and the reaction concentration of the antisense oligonucleotide combination of the CsABF8 gene is 100 μM.

[0035] The present invention also provides a method for constructing a drought-sensitive plant model, comprising the following steps:

[0036] The CsABF8 gene overexpression vector plasmid is introduced into Agrobacterium tumefaciens to obtain a recombinant bacterium; the recombinant bacterium is used to infect plants to obtain a drought-sensitive plant model.

[0037] In the specific implementation of the present invention, the backbone plasmid of the CsABF8 gene overexpression vector plasmid includes pCAMBIA 1305; the CsABF8 gene is inserted between the BamHI restriction site and the Spe I restriction site of pCAMBIA 1305.

[0038] The present invention also provides the use of the CsABF8 gene in regulating the raffinose content of plants; the amino acid sequence of the protein encoded by the CsABF8 gene is shown in SEQ ID NO.1.

[0039] In the present invention, the CsABF8 gene has the function of catalyzing the synthesis of raffinose in the tea plant. In the specific implementation of the present invention, the application includes the application of negatively regulating the CsABF8 gene to reduce the raffinose content in the plant or the application of positively regulating the CsABF8 gene to increase the raffinose content in the plant; the plant is the tea plant.

[0040] The present invention also provides the use of CsABF8 gene in regulating CsGolS1 / 2 promoter and / or CsRaf6 promoter; the amino acid sequence of the protein encoded by the CsABF8 gene is shown in SEQ ID NO.1.

[0041] In the present invention, after the CsABF8 gene is silenced, the expression levels of galactose pathway related genes CsGolS1 / 2 and CsRaf6 are significantly reduced; when the CsABF8 gene is overexpressed in the tea plant, the expression levels of the target genes CsGolS1 / 2 and CsRaf6 are significantly increased. In the specific implementation of the present invention, the application includes the application of negatively regulating the CsABF8 gene in reducing the gene expression mediated by the CsGolS1 / 2 promoter and / or the CsRaf6 promoter in the plant or the application of positively regulating the CsABF8 gene in increasing the gene expression mediated by the CsGolS1 / 2 promoter and / or the CsRaf6 promoter in the plant; the plant is a tea plant.

[0042] To further illustrate the present invention, the application of the CsABF8 gene provided by the present invention in regulating plant drought resistance is described in detail below in conjunction with the accompanying drawings and examples, but they should not be construed as limiting the scope of protection of the present invention.

[0043] Example 1

[0044] 1. Plant materials and processing

[0045] Two-year-old cutting seedlings of 'Fuyun No. 6' and 'Taicha No. 12' were purchased from Qianhe Tea Seedling Co., Ltd., Quanzhou, Fujian. Tea seedlings with a plant height of 30-35 cm, complete roots, and consistent overall growth were selected and hydroponically cultivated in the artificial climate room of Nongcui Garden, Anhui Agricultural University (greenhouse conditions: natural light, temperature: 20-25°C, humidity: 35%-40%). The nutrient solution was the nutrient solution of Xiaoxi Maoyi tea tree (pH adjusted to 4.5-5.5). Drought stress was simulated by applying 5% (g / ml) PEG (dissolved in the nutrient solution of Xiaoxi Maoyi tea tree), and samples of the second and third leaves of the tea seedlings at 0d, 1d, and 3d were collected and placed at -80°C. Some samples (control group and drought treatment 3d) were tested for non-targeted metabolomics, and the remaining samples were used for tea tree total RNA extraction for subsequent experiments.

[0046] One-year-old cuttings of 'Shuchazao' were purchased from Anhui Hefei Niansheng Agricultural Ecology Co., Ltd. and grown in a greenhouse using soil culture. All samples were collected and immediately frozen in liquid nitrogen and stored at -80°C until use.

[0047] 2. Determination of Raffinose Content in Tea Leaves

[0048] After the tea leaves are fully ground in liquid nitrogen, freeze-dried in a freeze dryer for 2 to 3 days. Take about 300 mg of freeze-dried tea leaves and soak them in 7 ml of ethanol with a volume concentration of 80% ethanol aqueous solution in an 80°C water bath for 30 minutes, shake 3 to 5 times for 10 minutes, centrifuge at 12000rpm for 5 minutes, collect the supernatant and incubate in a 95°C water bath until the ethanol evaporates. After cooling the remaining sugar solution to room temperature, put it at -80°C overnight and freeze-dry it in a freeze dryer for 2 to 3 days. The dry powder sugar mixture is dissolved in 500 μL Watsons water, shaken and mixed thoroughly, centrifuged at 12000rpm for 5 minutes, and the supernatant is collected. The raffinose content of tea leaves was detected using the plant raffinose (Raffinose) ELISA research kit provided by Jiangsu Enzyme Biotechnology Co., Ltd. The raffinose content was calculated using the following formula 1.

[0049] Raffinose (mg / gDW) = X × V / W × 10 -3 ×5 Formula 1;

[0050] In Formula 1, X is the sample concentration determined by the standard curve (μg / mL), V is the sample volume (μL), and W is the sample mass (g).

[0051] 3. Cloning of the CsABF8 gene from tea plant

[0052] 3.1 Cloning of CsABF8 gene

[0053] The CDS sequence of the CsABF8 gene was obtained using the tea plant genome website (http: / / tpia.teaplants.cn / index.html) (shown in SEQ ID NO. 2). The full-length cDNA of tea plant was used as a template and 2×Hieff PlusPCRMasterMix(With Dye)(Yisun Biotechnology (Shanghai) Co., Ltd.) high-fidelity enzyme premix was used for cloning of target genes, and the primers required for PCR amplification were CsABF8-F / R.

[0054] Primer sequences:

[0055] CsABF8-F: ATGAACTTCAAGAACTTCGG (SEQ ID NO.3);

[0056] CsABF8-R: TACCATGGCCCAGTCTGTG (SEQ ID NO. 4).

[0057] Table 1 Reaction system

[0058]

[0059]

[0060] Table 2 PCR reaction program

[0061]

[0062] The product amplified by PCR was connected to the pEASY-BluntZero cloning vector, and the connected vector was transformed into Escherichia coli DH5α competent cells. The full-length sequence of the CsABF8 gene was obtained by selecting monoclonal bacteria, colony PCR verification, and sending for comparison. The nucleotide sequence of the CsABF8 gene is shown in SEQ ID NO.2, and the amino acid sequence of the CsABF8 transcription factor is shown in SEQ ID NO.1.

[0063] 4. Transient silencing of the CsABF8 gene in tea leaves

[0064] (1) Submit the cloned tea tree CsABF8 gene sequence to the Soligo website (https: / / sfold.wadsworth.org / cgi-bin / soligo.pl), output the antisense oligonucleotide sequence and sense oligonucleotide sequence of the target gene, and input them into the TPIA (http: / / tpdb.shengxin.ren / index.html) website to screen out the nucleotide combination with strong specificity as the candidate sequence. Then send it to Shanghai Shenggong Biotechnology Co., Ltd. for sequence synthesis. The primer sequences are as follows:

[0065] Table 3 Silencing primer sequences of CsABF8

[0066]

[0067] (2) Dissolve the sense oligonucleotide and antisense oligonucleotide sequences of the synthesized silencing primer sequence in ultrapure water and dilute them to a final concentration of 100 μM;

[0068] (3) Take the tea tree cuttings (hydroponics) of 'Taicha No. 12' with the same growth conditions, and inject the sODN (sODN-CsABF8-1, sODN-CsABF8-2 and sODN-CsABF8-3 mixed in equal proportions) solution and AsODN (AsODN-CsABF8-1, AsODN-CsABF8-2 and AsODN-CsABF8-3 mixed in equal proportions) solution into the back of different leaves (use a 1mL syringe to absorb the diluted 100μM sODN or AsODN solution, gently rub the back of the second and third leaves of the plant with the syringe needle, and inject the silencing primer until the whole leaf is injected). After normal cultivation for 6h, treat with 15% PEG for 24h, take samples in batches and store them at -80℃ for later use, and then carry out the subsequent raffinose content determination and qRT-PCR gene quantitative test. See the schematic diagram for details. Figure 1 .

[0069] Through the transient silencing experiment in tea plants, the two and three leaves of tea hydroponic seedlings (TC) were treated with CsABF8 gene-specific sense oligonucleotides (sODN) or antisense oligonucleotides (AsODN) to achieve transient silencing of the CsABF8 gene in tea plants. qRT-PCR results showed that 6 hours after injection, AsODN treatment significantly inhibited the expression level of the CsABF8 gene compared with the control sODN treatment. qRT-PCR detection of galactose pathway-related genes CsGolS1 / 2 and CsRaf6 showed that the expression levels of the above genes decreased significantly after the CsABF8 gene was silenced. The tea plants that were successfully silenced were subjected to drought treatment for 24 hours, and it was found that the expression levels of CsABF8 in both the control group and the experimental group increased, indicating that the CsABF8 gene responded to drought stress, but the expression level of the experimental group was still lower than that of the control group, indicating that the expression of the gene was still inhibited at 24 hours of drought treatment. After 24 hours of drought treatment, the target genes CsGolS1 and CsGolS2 showed the same change trend as the CsABF8 gene, indicating that CsGolS1 and CsGolS2 were strongly regulated by the CsABF8 gene in the tea tree. The results of raffinose content showed that compared with the control group, the raffinose content in the leaves of the AsODN-CsABF8 treatment group was significantly reduced. After 24 hours of drought treatment, the raffinose content in both the sODN-CsABF8 control group and the AsODN-CsABF8 treatment group increased slightly, and was highly correlated with the CsABF8 gene expression results. The above results further confirmed that the CsABF8 gene has the function of catalyzing the synthesis of raffinose in the tea tree. See Figure 2 and Figure 3 .

[0070] Example 2

[0071] Transient overexpression of CsABF8 gene in tea plants

[0072] 1. Construction of CsABF8 gene overexpression vector

[0073] The pCAMBIA 1305 vector was double-digested with restriction endonucleases BamHI and Spe I to obtain a linearized vector. Recombinant primers were designed according to the CsABF8 gene sequence as follows:

[0074] CsABF8-1305-F:aggacagcccagatcactagtATGAACTTCAAGAACTTCGGAAATG (SEQ IDNO.11);

[0075] CsABF8-1305-R:gcccttgctcaccatggatccTCACCATGGCCCAGTCTGTG (SEQ IDNO.12);

[0076] According to the above-mentioned amplification system and amplification procedure, the target gene fragment was obtained, the target gene fragment was connected to the linearized vector using a one-step cloning method, the constructed pCAMBIA 1305-CsABF8 recombinant plasmid was transferred into Escherichia coli DH5α competent cells, a single clone was picked for colony PCR, the bacteria with the correct band were picked for sequencing, and after the sequence alignment was correct, the plasmid was extracted to obtain the pCAMBIA 1305-CsABF8 overexpression vector.

[0077] 2. Transient overexpression of tea tree

[0078] 1) Take out the competent cells of Agrobacterium tumefaciens GV3101 (pSoup) from the -80°C freezer, thaw on ice, and add no more than 1 μg of pCAMBIA-CsABF8 overexpression vector plasmid and pCAMBIA 1305 empty vector;

[0079] 2) Place the centrifuge tube in ice and let it stand for 5 minutes, then freeze it in liquid nitrogen for 5 minutes, bathe it in 37°C water for 5 minutes, and finally bathe it in ice for 5 minutes;

[0080] 3) Add 700 μL of liquid LB medium without antibiotics and culture at 28°C in a shaker at 220 rpm for 2 to 3 hours;

[0081] 4) Take 100 μL of bacterial solution and spread it on solid culture medium (containing 50 mg / L rifampicin (Rif) and 100 mg / L kanamycin (Kan) antibiotics), and incubate it upside down in a 28°C incubator for 2 to 3 days;

[0082] 5) Pick a single colony for colony PCR of the target gene, run gel electrophoresis and select the bacteria with the correct band for subsequent experiments;

[0083] 6) Add the bacteria carrying the pCAMBIA1305-CsABF8 recombinant vector and the bacteria containing the pCAMBIA 1305 empty vector to the liquid LB containing 50 mg / L Rif and 100 mg / L Kan antibiotics, and culture them in a shaking incubator at 28°C at 220 r / min until the OD 600 =0.8~1.0;

[0084] 7) Centrifuge at 5000 rpm for 10 min to collect the bacteria, and resuspend in MMA cell suspension to OD 600 =0.8~1.0, stand in the dark for 2h;

[0085] 8) Select soil-grown 'Shucha Zao' tea seedlings with the same growth status, and inject the dark-incubated mixed solution into tea leaves using a 1 mL syringe;

[0086] 9) After continuous cultivation at room temperature for 36 h, samples were collected and stored in a -80°C refrigerator for subsequent raffinose content determination and qRT-PCR gene quantitative testing.

[0087] Empty vectors pCAMBIA 1305 and pCAMBIA 1305-CsABF8 were overexpressed in tea plants, and the expression changes of genes related to the galactose pathway and the changes in the raffinose content in the tea plants were observed. The results showed that with the overexpression of the CsABF8 gene in the tea plants, the expression levels of the target genes CsGolS1 / 2 and CsRaf6 increased significantly, and the raffinose content in the tea leaves increased significantly. This once again verified that the CsABF8 gene in the tea plants increased the raffinose content by regulating the CsGolS1 / 2 and CsRaf6 promoters, thereby enabling the tea plants to cope with drought stress. Figure 4 and Figure 5 .

[0088] Although the above embodiment describes the present invention in detail, it is only a part of the embodiments of the present invention, not all of the embodiments. People can also obtain other embodiments based on this embodiment without creativity, and these embodiments all fall within the protection scope of the present invention.

Claims

1. Application of CsABF8 gene in regulating plant drought resistance; the amino acid sequence of the protein encoded by the CsABF8 gene is shown in SEQ ID NO.

1.

2. The use according to claim 1, characterized in that: The nucleotide sequence of the CsABF8 gene is shown in SEQ ID NO.

2.

3. The use according to claim 1, characterized in that: The application includes the application of negatively regulating the CsABF8 gene in improving plant drought resistance or the application of positively regulating the CsABF8 gene in constructing a drought-sensitive plant model.

4. The use according to claim 3, characterized in that: The negative regulation includes transient silencing; the reagent used for the negative regulation includes an antisense oligonucleotide combination targeting the CsABF8 gene; the antisense oligonucleotide combination includes the first antisense oligonucleotide shown in SEQ ID NO.8, the second antisense oligonucleotide shown in SEQ ID NO.9 and the third antisense oligonucleotide shown in SEQ ID NO.

10.

5. The use according to claim 3, characterized in that: The positive regulation includes transient overexpression; the reagent used in the positive regulation includes a CsABF8 gene overexpression vector plasmid; the CsABF8 gene overexpression vector plasmid uses pCAMBIA1305 as a backbone plasmid, and is inserted with the CsABF8 gene.

6. The use according to claim 1, characterized in that: The plants include tea plant.

7. A method for cultivating drought-resistant plants, characterized in that: The following steps are involved: The antisense oligonucleotide combination solution targeting the CsABF8 gene was injected into the mature leaves of the tea plant to achieve instantaneous silencing of the CsABF8 gene in the plant.

8. A method for constructing a drought-sensitive plant model, characterized in that: The following steps are involved: The CsABF8 gene overexpression vector plasmid is introduced into Agrobacterium tumefaciens to obtain a recombinant bacterium; the recombinant bacterium is used to infect plants to obtain a drought-sensitive plant model.

9. Application of CsABF8 gene in regulating the raffinose content of plants; the amino acid sequence of the protein encoded by the CsABF8 gene is shown in SEQ ID NO.

1.

10. Application of CsABF8 gene in regulating CsGolS1 / 2 promoter and / or CsRaf6 promoter; the amino acid sequence of the protein encoded by the CsABF8 gene is shown in SEQ ID NO.1.

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