Application of BnCYP90A1 gene in regulating rapeseed grain size and silique length

Abstract

The invention discloses the application of the BnCYP90A1 gene in regulating the seed size and silique length of rapeseed, belonging to the technical field of plant molecular biology. The invention for the first time discloses the application of the BnCYP90A1 gene of rapeseed in regulating the silique length and seed size of rapeseed, and the nucleotide sequence of the BnCYP90A1 gene is shown as SEQ ID NO.1. By overexpressing the BnCYP90A1 gene in the Brassica napus dm1-s1 mutant with reduced plant height, reduced seeds and shortened silique length, the invention constructs a complementation line and examines the silique length and 1000-seed weight of each line. The experimental results show that the BnCYP90A1 gene can significantly promote the elongation of siliques and the increase of seeds in rapeseed plants. The invention provides a new application direction for the BnCYP90A1 gene and provides new technical support for the breeding improvement of rapeseed seed yield and silique phenotype.

Description

Technical Field

[0001] The invention relates to the technical field of plant molecular biology, and in particular to application of a BnCYP90A1 gene in regulating rapeseed grain size and silique length. Background Art

[0002] Rapeseed, as one of the most important oil crops in my country, has important economic value, ornamental value and ecological value. With the growth of population, especially the continuous improvement of people's living standards, the supply and demand structure of edible vegetable oil in my country is unbalanced, the external dependence is getting higher and higher, and the oil security situation in my country is becoming increasingly severe. Limited by the influence of limiting factors such as land resources and planting costs in my country, the planting area of rapeseed in my country cannot be expanded on a large scale. Therefore, the increase in the yield of rapeseed per unit area is an important way to increase the yield of Brassica napus. Seed size and grain weight are significantly positively correlated and directly affect the yield of rapeseed per plant. Rapeseed silique is the organ where seeds are attached. It not only provides sufficient growth space for seed development, but also, in the late stage of seed maturity, as the main photosynthetic organ, it continuously provides nutrients for seed development to ensure the normal development of seeds. Changing the silique traits will directly or indirectly lead to yield changes. Therefore, improving the seed size and silique traits of rapeseed is of great significance to improving the yield of rapeseed.

[0003] Grain weight and silique length of rapeseed are important yield components at the individual level of rapeseed, and are also hot topics in the study of rapeseed yield traits. A large number of studies have been conducted to explore related genes and segments through QTL, and some genes have been cloned. For example, Auxin Response Factor 18 (ARF18) is a negative regulatory gene for silique length and grain weight of rapeseed. This gene controls silique length and seed size by inhibiting the expression of downstream auxin-related genes. The gene BnaA9.CYP78A9 encodes a P450 protein. After overexpression of this gene, rapeseed with longer siliques and increased grain weight was obtained. Some related genes have also been screened through reverse genetics. For example, overexpression of the Arabidopsis ubiquitin receptor mutant gene AtDA1R358K in rapeseed will lead to downregulation of endogenous BnDA1 expression in rapeseed, which significantly increases the seed size of rapeseed. At the same time, combined transcriptomics was used to identify genetic variation in the regulation of BnaUPL3.C03 in Brassica napus. BnaUPL3.C03 encodes HECTE3 ubiquitin ligase. After reducing the expression of BnaUPL3.C03, rapeseed seeds became significantly larger. Although researchers have cloned some genes that regulate seed size and silique length, a large number of genes have not been reported, and the specific pathways involved in their regulation and the key factors are still unknown. It is urgent to explore genes that have the function of regulating rapeseed seed size and silique length. Summary of the invention

[0004] The object of the present invention is to provide the application of the BnCYP90A1 gene in regulating the seed size and silique length of rapeseed, so as to solve the problems existing in the above-mentioned prior art. The BnCYP90A1 gene provided by the present invention can significantly promote the elongation of siliques and the increase of seeds in rapeseed plants.

[0005] To achieve the above object, the present invention provides the following solutions:

[0006] The present invention provides the application of the BnCYP90A1 gene in any one of the following:

[0007] (1) Application in regulating the seed size of rapeseed;

[0008] (2) Application in cultivating rapeseed varieties with large seeds;

[0009] (3) Application in regulating the silique length of rapeseed;

[0010] (4) Application in cultivating rapeseed varieties with long siliques;

[0011] The nucleotide sequence of the BnCYP90A1 gene is shown as SEQ ID NO.1.

[0012] The present invention provides the application of the recombinant vector comprising the BnCYP90A1 gene in any one of the following:

[0013] (1) Application in regulating the seed size of rapeseed;

[0014] (2) Application in cultivating rapeseed varieties with large seeds;

[0015] (3) Application in regulating the silique length of rapeseed;

[0016] (4) Application in cultivating rapeseed varieties with long siliques.

[0017] The present invention provides the application of the engineering bacteria comprising the recombinant vector in any one of the following:

[0018] (1) Application in regulating the seed size of rapeseed;

[0019] (2) Application in cultivating rapeseed varieties with large seeds;

[0020] (3) Application in regulating the silique length of rapeseed;

[0021] (4) Application in cultivating rapeseed varieties with long siliques.

[0022] The present invention also provides the application of the BnCYP90A1 protein in any one of the following:

[0023] (1) Use in regulating rapeseed grain size;

[0024] (2) Use in cultivating rapeseed varieties with large grains;

[0025] (3) Use in regulating the silique length of rapeseed;

[0026] (4) Use in cultivating rapeseed varieties with long siliques;

[0027] The amino acid sequence of the BnCYP90A1 protein is shown in SEQ ID NO.2.

[0028] Preferably, up-regulating the expression level of the BnCYP90A1 gene promotes the enlargement of rapeseed grains and / or promotes the elongation of rapeseed siliques.

[0029] The present invention also provides a method for promoting the enlargement of rapeseed grains, which includes the step of overexpressing the BnCYP90A1 gene in rapeseed, and the nucleotide sequence of the BnCYP90A1 gene is shown in SEQ ID NO.1.

[0030] The present invention also provides a method for promoting the elongation of rapeseed siliques, which includes the step of overexpressing the BnCYP90A1 gene in rapeseed, and the nucleotide sequence of the BnCYP90A1 gene is shown in SEQ ID NO.1.

[0031] Preferably, the overexpression of the BnCYP90A1 gene in rapeseed includes the following steps: constructing a recombinant plasmid by ligating the BnCYP90A1 gene with an expression vector; transferring the recombinant plasmid into rapeseed by Agrobacterium-mediated transformation to overexpress the BnCYP90A1 gene.

[0032] The present invention also provides a method for cultivating rapeseed with large grains, which includes the step of overexpressing the BnCYP90A1 gene in rapeseed, and the nucleotide sequence of the BnCYP90A1 gene is shown in SEQ ID NO.1.

[0033] The present invention also provides a method for cultivating rapeseed with long siliques, which includes the step of overexpressing the BnCYP90A1 gene in rapeseed, and the nucleotide sequence of the BnCYP90A1 gene is shown in SEQ ID NO.1.

[0034] The present invention discloses the following technical effects:

[0035] The present invention discloses for the first time the application of the rapeseed BnCYP90A1 gene in regulating the silique length and seed size of rapeseed. By overexpressing the BnCYP90A1 gene in the Brassica napus dm1-s1 mutant with reduced plant height, reduced seeds and shortened silique length, a complementation line was constructed, and the silique length and 1000-grain weight of each line were investigated. The experimental results show that the BnCYP90A1 gene can significantly promote the elongation of siliques and the enlargement of seeds in rapeseed plants. The present invention provides a new application direction for the BnCYP90A1 gene and provides new technical support for the breeding improvement of rapeseed seed yield and silique phenotype. Brief Description of the Drawings

[0036] In order to more clearly illustrate the technical solutions in the embodiments of the present invention or the prior art, the following will briefly introduce the drawings required for use in the embodiments. Obviously, the drawings in the following description are only some embodiments of the present invention. For those of ordinary skill in the art, without creative efforts, other drawings can be obtained based on these drawings.

[0037] Figure 1 It is a phenotypic diagram of the dm1-s1 mutant plant and Brassica napus ZS11; among them, a is the overall phenotypic appearance of the plant, and the scale bar is 10 cm; b is the silique phenotype, and the scale bar is 1 cm; c is the statistical chart of seed weight, Thousand grain weight is the 1000-grain weight, dm1 is the dm1-s1 mutant plant, and *** indicates P<0.001;

[0038] Figure 2 It is a phenotypic diagram of the dm1-s1 mutant plant, Brassica napus ZS11, and the BnCYP90A1 transgenic positive plants com.#1 and com.#2 at the seedling stage; the scale bar in the figure is 10 cm;

[0039] Figure 3 It is a morphological phenotypic diagram of the siliques of the dm1-s1 mutant plant, Brassica napus ZS11, and the BnCYP90A1 transgenic positive plants com.#1 and com.#2; the scale bar in the figure is 5 cm;

[0040] Figure 4 It is a statistical analysis diagram of the silique length of the dm1-s1 mutant plant, Brassica napus ZS11, and the BnCYP90A1 transgenic positive plants com.#1 and com.#2; among them, Silique length is the silique length, ns indicates no significant difference, * indicates P<0.05, and **** indicates P<0.0001;

[0041] Figure 5Statistical analysis chart of 1000-grain weight of dm1-s1 mutant plants, Brassica napus ZS11, and BnCYP90A1 transgenic positive plants com.#1 and com.#2; among them, Thousand grain weight represents the 1000-grain weight, and *** indicates P<0.001. Detailed implementation mode

[0042] The various exemplary embodiments of the present invention will now be described in detail. This detailed description should not be considered as a limitation of the present invention, but rather as a more detailed description of certain aspects, characteristics, and implementation schemes of the present invention.

[0043] It should be understood that the terms used in the present invention are only for describing specific embodiments and are not used to limit the present invention. Additionally, for the numerical ranges in the present invention, it should be understood that each intermediate value between the upper and lower limits of the range is also specifically disclosed. Any intermediate value within any stated value or stated range, as well as each smaller range between any other stated value or intermediate value within the stated range, is also included in the present invention. The upper and lower limits of these smaller ranges can be independently included or excluded from the range.

[0044] Unless otherwise specified, all technical and scientific terms used herein have the same meaning as commonly understood by those of ordinary skill in the art to which the present invention pertains. Although the present invention only describes preferred methods and materials, any methods and materials similar or equivalent to those described herein can also be used in the implementation or testing of the present invention. All documents mentioned in this specification are incorporated by reference to disclose and describe the methods and / or materials related to the documents. In case of conflict with any incorporated document, the content of this specification shall prevail.

[0045] Without departing from the scope or spirit of the present invention, various improvements and changes can be made to the specific implementation modes of the present invention specification, which are obvious to those skilled in the art. Other implementation modes obtained from the present invention specification are obvious to those skilled in the art. The present invention specification and examples are only exemplary.

[0046] Regarding the use of "comprising", "including", "having", "containing", etc. in this article, they are all open-ended terms, meaning including but not limited to.

[0047] The overall experimental concept of the embodiments of the present invention is as follows:

[0048] Cloning of BnCYP90A1 gene: Using the DNA and RNA of Brassica napus ZS11 as templates respectively, and amplifying with primers BnCYP90A1-F / BnCYP90A1-R and primers pBnCYP90A1-F / pBnCYP90A1-R to obtain the promoter of BnCYP90A1 and the coding region fragment of BnCYP90A1. Then, the above fragments were fused with the pCAMBIA1300 vector by homologous recombination and transferred into Escherichia coli DH5α to screen for positive clones.

[0049] Agrobacterium-mediated genetic transformation: The obtained positive plasmid was transferred into Agrobacterium tumefaciens GV3101, and the hypocotyl of dm1-s1 was used as the infection site to transfer BnCYP90A1 into the mutant material. After screening with hygromycin, independent resistant calli were obtained and further differentiated and regenerated into complete transgenic lines.

[0050] Application of BnCYP90A1 gene: By comparing the silique length and 1000-seed weight of dm1-s1 and the complementation material, it was found that compared with dm1-s1, the silique length and 1000-seed weight of the complementation material were much greater than those of dm1-s1.

[0051] The method for preparing the culture media used in the examples of the present invention is as follows:

[0052] M1 solid medium: Weigh 4.4 g of MS, 30 g of sucrose, 18 g of mannitol, 5.5 g of phytagel, make up to 1 L of aqueous solution, after autoclaving, add 1 mg of 2,4-dichlorophenoxyacetic acid, 0.3 mg of kinetin, 5.5 g of phytagel and 100 μmol of acetosyringone that are filter-sterilized, and mix evenly to prepare a plate.

[0053] M2 solid medium: Weigh 4.4 g of MS, 30 g of sucrose, 18 g of mannitol, 5.5 g of phytagel, make up to 1 L, after autoclaving, supplement 1 mg of 2,4-dichlorophenoxyacetic acid, 0.3 mg of kinetin, 300 mg of ticarcillin, 150 μmol of sodium thiosulfate and 50 mg of hygromycin that are filter-sterilized, and mix evenly to prepare a plate

[0054] M3 solid medium: Weigh 4.4 g of MS, 10 g of glucose, 0.25 g of xylose, 0.6 g of 2-(N-morpholino)ethanesulfonic acid, make up to 1 L, add 5.5 g of phytagel, after autoclaving for 20 min, wait for the medium to cool and then add 2 mg of zeatin, 0.1 mg of indoleacetic acid, 300 mg of ticarcillin, 150 μmol of silver nitrate that are filter-sterilized in sequence, and mix evenly to prepare a plate.

[0055] M4 medium: Weigh 4.4 g of MS and 10 g of sucrose, dissolve them in water, make up the volume to 1 L, and finally add 5.5 g of phytagel, then sterilize.

[0056] DM liquid medium: Weigh 4.4 g of MS and 30 g of sucrose, dissolve them in 1 L of medium, sterilize, and then add 100 μmol of filter-sterilized acetosyringone.

[0057] Table 1 Sequences of primers used in the examples of the present invention

[0058]

[0059] The Brassica napus ZS11 (Zhongshuang No. 11) used in the examples of the present invention was cultivated by the Oil Crops Research Institute of the Chinese Academy of Agricultural Sciences in Wuhan, and has been publicly reported in the literature "Wu Yongcheng, Ma Ni, Huang Xiaoming, etc. Effects of nitrogen application rate on agronomic traits, yield, quality and nitrogen use efficiency of Brassica napus cv. Zhongshuang 11 [J]. Journal of Sichuan Agricultural University, 201, 32(03): 260-264+282." The applicant undertakes to make the above biological materials available to the public within twenty years from the filing date of the present invention.

[0060] The Brassica napus dm1-s1 mutant material used in the examples of the present invention was obtained by EMS mutagenesis of the excellent cultivated variety ZS11 of Brassica napus. The applicant undertakes to make the above biological materials available to the public within twenty years from the filing date of the present invention.

[0061] As Figure 1 shown, compared with Brassica napus ZS11, the dm1-s1 mutant showed shorter plant height, shorter silique length and significantly smaller seeds.

[0062] Example 1 Cloning of the promoter of Brassica napus BnCYP90A1 and the BnCYP90A1 coding region fragment

[0063] 1. Extract the RNA and DNA of Brassica napus ZS11 using the Trizol method and the CTAB method respectively

[0064] RNA extraction by the Trizol method: Collect the seedlings of Brassica napus ZS11, after sufficient fragmentation with liquid nitrogen, add Trizol solution (Invitrogen, 15596026CN), vortex and mix evenly, then add chloroform, and react on ice for 5-10 min. Subsequently, centrifuge and aspirate the supernatant, mix it with an equal volume of isopropanol to precipitate RNA. Finally, rinse the RNA with absolute ethanol, air dry at room temperature, and add RNase-free water to obtain the RNA of Brassica napus ZS11.

[0065] DNA extraction by CTAB method: Small seedlings of Brassica napus ZS11 were collected, added with CTAB solution for disruption, and placed in an incubator at 65 °C for 1 hour. Subsequently, a chloroform / isopentanol mixture was added, and after mixing evenly, centrifugation was performed and the supernatant was transferred and mixed with an equal volume of absolute ethanol. After white precipitate was precipitated, the precipitate was retained, rinsed twice, and double-distilled water was added to obtain DNA.

[0066] 2. Obtaining the promoter and coding region fragment of BnCYP90A1 by PCR amplification

[0067] Using the reverse transcription kit (R423-01, Vazyme) of Novizan, reverse transcribe the RNA of Brassica napus ZS11 small seedlings according to the steps in the instruction manual to obtain ZS11 cDNA, and dilute the obtained cDNA product by 3 times as the PCR amplification template. For the amplification of the BnCYP90A1 promoter, 1 μl of ZS11 DNA was taken as the template, and the amplification primers pBnCYP90A1-F / pBnCYP90A1-R (the primer sequences are shown in Table 1, and the lowercase letters are homologous arm primers) were added for PCR reaction. For the amplification of the BnCYP90A1 coding region fragment, 1 μl of ZS11 cDNA was taken as the template, and the primers BnCYP90A1-F / BnCYP90A1-R (the primer sequences are shown in Table 1, and the lowercase letters are homologous arm primers) were added for amplification reaction. After the reaction ended, the amplification products of the BnCYP90A1 promoter and BnCYP90A1 coding region fragment were recovered by agarose gel electrophoresis; the amplification products were collected separately, purified, and sent to Shanghai Sangon Biological Engineering Co., Ltd. for sequencing to obtain the nucleotide sequence of the gene coding region (CDS sequence) of the BnCYP90A1 gene, as shown in SEQ ID NO.1; the amino acid sequence of the BnCYP90A1 protein can be obtained, as shown in SEQ ID NO.2; the nucleotide sequence of the BnCYP90A1 promoter, as shown in SEQ ID NO.3.

[0068] SEQ ID NO.1:

[0069]

[0070] SEQ ID NO.2:

[0071] MAFSFSSTALLLLLSSITAGFLLLLRRTRYRRMGLPPGSLGLPLIGETLQLIAAYKTENPEPFVDARVARYGSVFMTHIFGEPTVFSADPETNRFVLQNEGKLFECSYPASICNLLGKHSLLLMKGSLHKRMHSLTMSFANSSIIKDHLMLDIDRLVRFNLDSWSSRVLLMEEAKKITFELTVKQLMSFDPGEWSESLRKEYLLVIEGFFSIPLPLFSTTYRKAIKARTKVAEALTVVVMKRREEEEEGEERKKDMLAALLAAEERFSDEEIVDFLVALLVAGYETTSTIMTLAVKFLTETPSALAQLKEEHENIRAMKSDSYSLEWSDYKSMPFTQCVVSETLRVANIIGGVFRRATTDVEIKGYKIPKGWKVFASFRAVHLDPNHFKDARTFNPWRWQGNSVTTCPSNVFTPFGGGPRLCPGYELARVALSVFLHRLVTGFSWVPAEKDKLVFFPTTRTQKRYPIIVTRRDDGGLSAT。

[0072] SEQ ID NO.3:

[0073]

[0074] Example 2 Construction of Overexpression Vector

[0075] 1. Homologous Recombination of Overexpression Vector

[0076] Extract the plasmid expression vector pCAMBIA1300-pUBQ10-flag (the sequence of the expression vector pCAMBIA1300-pUBQ10-flag has been disclosed in Patent CN118127038A), and linearize the vector using the restriction enzymes EcoRI and SpeI. Then, use the Novoprotein homologous recombination kit (C116-01, Novoprotein) to perform homologous recombination according to the instructions of the manual to obtain the recombinant product.

[0077] 2. Transformation of Escherichia coli with Recombinant Product and Screening of Positive Clones

[0078] Add the recombinant product to the Escherichia coli competent cell DH5α, and transfer the recombinant product into Escherichia coli by heat shock amplification. The next day, use the identification primers (BnCYP90A1-GT-F and BnCYP90A1-GT-R) to identify each monoclonal grown one by one. For the obtained positive clones, expand and extract the plasmid using the Thermo Fisher Scientific plasmid extraction kit (K0503, Thermo Fisher Scientific), sequence it, and finally obtain the pCAMBIA1300-pBnCYP90A1-BnCYP90A1-flag plasmid.

[0079] Example 3 Genetic Transformation of Rapeseed

[0080] Sterilized ZS11 is placed in a germination box for germination. During seed germination, the target plasmid is transferred into Agrobacterium tumefaciens GV3101 to obtain the Agrobacterium tumefaciens required for transformation. After the hypocotyls of ZS11 seedlings germinate to an appropriate length, cut off the hypocotyls and co-incubate them with the Agrobacterium tumefaciens solution resuspended in DNA liquid. After the incubation, transfer the hypocotyls to M1 medium for dark culture. After 2 days, transfer the hypocotyls to M2 medium and culture for 15 days, and continue to culture on M3 medium until budding. After budding, transfer to M4 medium until rooting. When the seedlings grow to an appropriate size, transplant them into the soil for continued cultivation. The obtained transgenic positive plants (complemented lines) com.#1, com.#2 are as Figure 2 shown.

[0081] Example 4 Identification of Positive Plants

[0082] Collect single leaves of rapeseed plants, quickly freeze them in liquid nitrogen, and extract DNA using the CTAB method. Design PCR primers BnCYP90A1-GT-F and BnCYP90A1-GT-R (the primer sequences are shown in Table 1) using Primer Design software. Use the extracted transgenic rapeseed DNA as a template for amplification, and then analyze it by DNA gel electrophoresis to confirm positive plants.

[0083] Example 5 Determination of Rapeseed Silique Length and 1000-Grain Weight

[0084] Transplant the above transgenic materials to a transgenic nursery for cultivation. There are 5 holes in each row in the nursery, with a hole spacing of 20 cm, 3 plants in each hole, and 5 rows are transplanted for each transgenic line, with a row spacing of 33 cm. After the transgenic materials grow naturally to maturity, collect 30 siliques from the main inflorescence for silique length measurement. At the same time, determine the 1000-grain weight of the harvested rapeseed grains.

[0085] The results are as Figures 3 - 5 shown. Compared with the mutant line dm1-s1, the silique lengths of the plants in the complementation lines com.#1 and com.#2 are significantly increased, and the 1000-grain weight is significantly increased. This shows that BnCYP90A1 can significantly promote the development of rapeseed siliques and the enlargement of seeds.

[0086] The above embodiments are only descriptions of the preferred embodiments of the present invention, and do not limit the scope of the present invention. Without departing from the design spirit of the present invention, various deformations and improvements made by those of ordinary skill in the art to the technical solutions of the present invention shall fall within the protection scope determined by the claims of the present invention.

Claims

1. Use of overexpressing the BnCYP90A1 gene in any of the following: (1) Use in promoting the enlargement of rapeseed grains; (2) Use in cultivating rapeseed varieties with large grains; (3) Use in promoting the elongation of rapeseed pods; (4) Use in cultivating rapeseed varieties with long pods; The nucleotide sequence of the BnCYP90A1 gene is shown as SEQ ID NO.

1.

2. Use of the overexpression vector comprising the BnCYP90A1 gene described in claim 1 in any of the following: (1) Use in promoting the enlargement of rapeseed grains; (2) Use in cultivating rapeseed varieties with large grains; (3) Use in promoting the elongation of rapeseed pods; (4) Use in cultivating rapeseed varieties with long pods.

3. Use of the engineered bacteria comprising the overexpression vector described in claim 2 in any of the following: (1) Use in promoting the enlargement of rapeseed grains; (2) Use in cultivating rapeseed varieties with large grains; (3) Use in promoting the elongation of rapeseed pods; (4) Use in cultivating rapeseed varieties with long pods.

4. Use of overexpressing the BnCYP90A1 protein in any of the following: (1) Use in promoting the enlargement of rapeseed grains; (2) Use in cultivating rapeseed varieties with large grains; (3) Use in promoting the elongation of rapeseed pods; (4) Use in cultivating rapeseed varieties with long pods; The amino acid sequence of the BnCYP90A1 protein is shown as SEQ ID NO.

2.

5. A method for promoting the enlargement of rapeseed grains, characterized in that, Including the step of overexpressing the BnCYP90A1 gene in rapeseed, and the nucleotide sequence of the BnCYP90A1 gene is shown as SEQ ID NO.

1.

6. A method for promoting the elongation of rapeseed pods, characterized in that, Including the step of overexpressing the BnCYP90A1 gene in rapeseed, and the nucleotide sequence of the BnCYP90A1 gene is shown as SEQ ID NO.

1.

7. The method according to claim 5 or 6, characterized in that, The step of overexpressing the BnCYP90A1 gene in rapeseed includes the following steps: constructing a recombinant plasmid by ligating the BnCYP90A1 gene with an expression vector; transferring the recombinant plasmid into rapeseed by Agrobacterium-mediated transformation to overexpress the BnCYP90A1 gene.

8. A method for cultivating large-seeded rapeseed, characterized in that, Including the step of overexpressing the BnCYP90A1 gene in rapeseed, and the nucleotide sequence of the BnCYP90A1 gene is shown as SEQ ID NO.

1.

9. A method for cultivating Brassica napus var. megacarpa, characterized in that, Including the step of overexpressing the BnCYP90A1 gene in rapeseed, and the nucleotide sequence of the BnCYP90A1 gene is shown as SEQ ID NO.1.

Images