Tumorous stem BjuSPL10c gene for regulating and controlling swelling of tumorous stem, overexpression vector, construction method and application of tumorous stem BjuSPL10c gene

Abstract

The invention belongs to the technical field of plant molecular breeding, and particularly relates to a tumorous stem mustard BjuSPL10c gene for regulating and controlling expansion of tumorous stems, an overexpression vector, a construction method and application of the tumorous stem mustard BjuSPL10c gene. The nucleotide sequence of the tumorous stem mustard BjuSPL10c gene for regulating and controlling expansion of tumorous stems is shown as SEQ ID NO.1, and the amino acid sequence of the tumorous stem mustard BjuSPL10c gene is shown as SEQ ID NO.2. According to the invention, the full-length sequence of the BjuSPL10c gene of the tumorous stem mustard is cloned for the first time, and a protein sequence coded by the BjuSPL10c gene is obtained. By constructing an overexpression vector, a tumorous stem mustard plant with the overexpressed BjuSPL10c gene is obtained. The overexpression of the tumorous stem mustard BjuSPL10c can promote the expansion of the tumorous stem of the tumorous stem mustard, so that an effective gene is provided for the genetic engineering of the tumorous stem mustard and other plants. The invention also provides an important target for molecular genetic improvement of metamorphosis stem development plants, and has important value and significance for cultivation and production of a new variety of stem nodule mustard.

Description

Technical Field

[0001] The invention belongs to the technical field of plant molecular breeding, and specifically relates to a tumor-bearing mustard BjuSPL10c gene for regulating tumor expansion, an overexpression vector, a construction method and applications thereof. Background Art

[0002] Tuberous stem mustard, Brassica juncea var.Tumida Tsen et Lee, is not only a fresh vegetable, but also an important pickled and processed vegetable. The metamorphosed tubers of Brassica juncea are important organs for fresh eating and processing, and are the main raw materials and breeding traits for making Fuling Zhacai. Therefore, tubers are the main target traits of Brassica juncea breeding. Tuberous stems of Brassica juncea are aboveground metamorphosed stems. The 1 to 5 tumor-like protrusions formed by the metamorphosis of the petiole base are formed with the lateral expansion of the stem. There are few reports on the relevant molecular regulatory mechanisms. Previous studies have found that the expansion of Brassica juncea tubers is a process of increasing the number of cells and increasing the volume. The cambium cells of Brassica juncea first divide tangentially, then divide radially to form the long axis of the tuber, and finally form an expanded tuber. In addition, nuclear polyploidy is also involved in the expansion of Brassica juncea tubers. The external myelocytes are the source of the increase in the number of cells in the expanded tuber, while nuclear re-replication is involved in the expansion of the volume of the internal myelocytes.

[0003] SPLs are plant-specific transcription factors that contain a highly conserved SBP functional domain consisting of 76 amino acid residues, which acts on nuclear import and sequence-specific binding to flanking regions containing the GTAC core motif and gene-specificity. Studies have found that there are 17 SPLs genes in Arabidopsis, whose proteins contain a highly conserved DNA-binding domain, SBP-DBD and two zinc-finger like structures consisting of 79 amino acid residues, and are involved in various stages of plant growth and development. Literature reports that 10 SPLs members in Arabidopsis are downregulated by miR156. According to the different functions of these SPLs, they can be divided into three categories: 1. Regulating the transition from juvenile to adult vegetative growth period and from vegetative growth period to reproductive growth period, including SPL2, SPL9, SPL10, SPL11, SPL13 and SPL15; 2. Promoting the transition of floral meristem, including SPL3, SPL4 and SPL5; 3. Unknown functions, including SPL16. In addition, SPL1 and SPL12 are involved in regulating the heat tolerance of plants during their reproductive growth period, SPL8 is involved in GA-regulated plant development, SPL6 is involved in plant immune response, SPL7 is involved in regulating copper homeostasis within plants, and SPL14 is involved in regulating plant morphological construction.

[0004] In summary, the problem existing in the prior art is that although the SPL10 gene of Arabidopsis thaliana has been studied, there is no research report on the function of the BjuSPL10c gene of Tuber mustard, and its specific function is still unclear. Summary of the invention

[0005] The purpose of the present invention is to provide a BjuSPL10c gene for regulating tumor expansion of stem mustard, an overexpression vector, a construction method and an application thereof, and a gene that can directly regulate the development of tumors of stem mustard is discovered.

[0006] The technical solution adopted by the present invention is a BjuSPL10c gene for regulating tumor expansion, the nucleotide sequence of which is shown in SEQ ID NO.1; and the amino acid sequence of which is shown in SEQ ID NO.2.

[0007] SEQ ID NO.1:

[0008] 5-’ATGGACTGCAACATGGTATCTCCGATGTGGGACTGGGAGCATTTGA TCATCCCCAACCTGTCAAAGACTGAAACTGACAAGAAACCGCCTTCTACTACTGATTGGGATATTGAGAAAGGTGAAGGGATTGAATCTATATTATTTCCCTCTTTCACTGGCTCCTCCTCCACCACCGGTTTCCTAAGCTCACAGTTGACCTCCACCAACTCATCATCTCCCAAACTCAAAAGTTCCCTTGGAGATTTTGACCAAGTCAAGGCATCCACAGCTCTCCAGGTGGTATCTGCTGAATCAGACCTTTGTTTAAAACTTGGGAAGCGGAGTTACTCTGATGAAGAGACTTGGGGTAGAAACAACAATAATGACATTTCAGCCGCTGTTTCTGTGAAGCTCTTGACTCCTCCATCTGTTGTTGCTCTGAAGAAATCCAAATCATCATGTGGTCAGAGCATGCAAGTACCTCGTTGTCAGATTGATGGCTGTGAACTGGATCTCTCATCTGCCAAGGATTATCATCGCAAGCATAGAGTCTGCGAACGCCATTCAAAGTGCCCTGTAGTTACTGTGGGTGGCTTTGAACGTCGATTCTGCCAACAGTGTAGCAGGTTGCATGCGGTCTCTGAATTTGATGAGAAGAAACGTAGCTGCCGTAAACGTCTTTCGCATCATAATGCGAGGCGCCGCAAGTCACAAGGAATGTTTCCTTTGAATCCAGAGAGGGTGTACAATGGAAGACAGCATACGAATATGTTGTGGAATGAGTTGTCCCTTAACACAGGATCTGAAGAAAAGTATGCATGGGGTACTACTACTTATGAGACAAAGCCTACGCAGATGGAAAGCGGCTTTACTCTGAACTTCCAGAGAGGTAGTGGCGCTGAGGAGCAGCAGGTGTTTGCTAGTAGCAACCTTTCTTTCTCTGCGTATCAACCCTCAGCAGGGAAATCCAACGTTCAACTTCCTAGCAAAGGTGTAGGAGAATACTCAGGAG GCCTCTATGAAACGCAAGATTTCTACAGTGCTCTCTCTCTTCTGTCAACGTCTTCGGATTCACATGGGACCAAACACCATCCCATGGTTGAATCACAGCCAATACATGGCACCTTCCCTACTCATTTTTTATGA-3'.

[0009] SEQ ID NO.2:

[0010] MDCNMVSPMWDWEHLIIPNLSKTETDKKPPSTTDWDIEKGEGIESILFPSFFTGSSSTTGFLSSQLTSTNSSSPKLKSSLGDFDQVKASTALQVVSAESDLCLKLGKRSYSDEETWGRNNNNNDISAAVSVKLLTPPSVVALKKSKSSCGQSMQVPRCQIDGCELDLSSAKDYHRKHRVCERHSKCPVVTVGGFE RRFCQQCSRLHAVSEFDEKKRSCRKRLSHHNARRRKSQGMFPLNPERVYNGRQHTNMLWNELSLNTGSEEKYAWGTTTYETKPTQMESGFTLNFQRGSGAEEQQVFASSNLSFSAYQPSAGKSNVQLPSKGVGEYSGGLYETQDFYSALSLLSTSSDSHGTKHHPMVESQPIHGTFPTHFL*, where * represents the termination signal.

[0011] An overexpression vector constructed using the BjuSPL10c gene of stem tumor mustard.

[0012] An overexpression vector constructed using the stem tumor mustard BjuSPL10c gene, the construction method of which comprises the following steps: synthesizing a pair of primers, the primer sequences are shown in SEQ ID NO.3 and SEQ ID NO.4; extracting total RNA from wild-type stem tumor mustard tuber, and reverse transcribing it into cDNA; using the cDNA as a template, performing PCR amplification with the primers to obtain the stem tumor mustard BjuSPL10c target gene; and sequentially performing enzyme digestion, connection and transformation on the expression vector and the stem tumor mustard BjuSPL10c target gene to obtain the overexpression vector.

[0013] Preferably, the tuber is a tuber in the middle stage of expansion.

[0014] Preferably, the expression vector is p1300-GFP.

[0015] Application of the BjuSPL10c gene or an overexpression vector of the BjuSPL10c gene in promoting the enlargement of tumor stems of the BjuSPL10c gene.

[0016] The overexpression vector is used to prepare a tumorous mustard plant that overexpresses the BjuSPL10c gene, and the preparation process comprises the following steps: the overexpression vector is used to transform Agrobacterium competent cells to obtain positive recombinant Agrobacterium; the positive recombinant Agrobacterium is used to infect the cotyledons of the tumorous mustard seedlings, and the cotyledons of the tumorous mustard seedlings are transferred to a co-culture medium and cultured in the dark; the cotyledons of the tumorous mustard seedlings are sequentially transferred to a callus induction culture medium to produce callus, an adventitious bud differentiation culture medium to produce adventitious buds, and a rooting culture medium to produce roots, so as to obtain a tumorous mustard plant that overexpresses the BjuSPL10c gene.

[0017] Preferably, when the positive recombinant Agrobacterium infects the cotyledons of the tumorous mustard seedlings, the bacterial solution OD 600 is 0.05.

[0018] Preferably, the callus induction medium formula is: add 2 mg of 6-benzylaminopurine, 0.15 mg of α-naphthylacetic acid, 20 mg of hygromycin, 500 mg of timentin and 8 g of agar per liter of MS medium.

[0019] The formula of adventitious bud differentiation medium is: add 2 mg of 6-benzylaminopurine, 0.15 mg of α-naphthylacetic acid, 20 mg of hygromycin, 500 mg of timentin and 8 g of agar per liter of MS medium.

[0020] The formula of the rooting medium is as follows: add 0.1 mg of α-naphthylacetic acid, 20 mg of hygromycin, 500 mg of timentin and 8 g of agar per half liter of MS medium.

[0021] The beneficial effects of the present invention are that the present invention cloned the full-length sequence of the stem tumor mustard BjuSPL10c gene for the first time, obtained the protein sequence encoded by the gene, and obtained the stem tumor mustard plant overexpressing the BjuSPL10c gene by constructing an overexpression vector. The results show that overexpression of the BjuSPL10c gene can promote the enlargement of the stem tumor mustard. This provides an effective gene for genetic engineering of stem tumor mustard and other plants. The present invention provides an important target for molecular genetic improvement of plants with abnormal stem development, and has important value and significance for the cultivation and production of new varieties of stem tumor mustard. BRIEF DESCRIPTION OF THE DRAWINGS

[0022] Figure 1 This is the electrophoresis diagram of target gene amplification and restriction enzyme digestion of the expression vector of B.

[0023] Figure 2 This is the electrophoresis diagram of gDNA detection results in tumorous mustard plants overexpressing the BjuSPL10c gene.

[0024] Figure 3 This is a graph showing the results of detecting the expression level of BjuSPL10c mRNA in tumorous mustard plants that overexpress the BjuSPL10c gene.

[0025] Figure 4 Comparison results of tumors between tumorous mustard plants overexpressing the BjuSPL10c gene and wild-type tumorous mustard plants; a is the tumor weight statistics; b is the phenotypic diagram. DETAILED DESCRIPTION

[0026] The present invention is further described below by specific examples, but the scope of the present invention is not limited thereto. The details and forms of the technical solution of the present invention may be modified or replaced without departing from the spirit and scope of the present invention, but these modifications or replacements all fall within the protection scope of the present invention.

[0027] The inventive concept of the present invention is as follows: The present invention provides a BjuSPL10c gene for regulating tumor expansion, wherein the nucleotide sequence of the BjuSPL10c gene is shown in SEQ ID NO.1; and the amino acid sequence is shown in SEQ ID NO.2.

[0028] The inventors discovered the BjuSPL10c gene and overexpressed the BjuSPL10c gene in tumorous stem mustard to promote the expansion of plant tubers and increase the yield of tumorous stem mustard. Tumorous stem mustard is a biennial crop harvested in the vegetative period. The base of its petiole is transformed into 1-5 tumor-like protrusions, which are formed with the lateral expansion of the stem. The tubers are the main harvesting organs, involving many factors such as morphology, physiology, biochemistry and genetics. Morphological studies have found that cell division and nuclear re-replication are closely related to tuber expansion, among which the division of external myelocytes increases the number of cells in the expanded tuber, while nuclear re-replication promotes the expansion of internal myelocytes. The expansion of tumors in tumorous stem mustard is always accompanied by the expansion of cell volume. First, the cambium cells divide tangentially to increase the number of cell layers, and then radial division forms the long axis of the tuber, followed by non-directional division in the tuber, and the tuber develops in all directions, finally forming an expanded tuber. Previous studies have found that there is nuclear polyploidization during the process of tuber expansion, the cell ploidy continues to increase, the degree of nuclear polyploidy increases, and the cell ploidy decreases first and then increases from the outside to the inside of the mature stem, among which the cell ploidy near the cambium is the lowest and the inner layer of the pith is the highest. In-depth studies have found that BjXTH1, xyloglucanendotransglucosylase / hydrolase 1, BjXTH2, xyloglucan endotransglucosylase / hydrolase 2, BjuBXL1-1, β-D-xylosidase 1-1 and BjuBXL1-2, β-D-xylosidase 1-2 are involved in regulating the expansion of pith cells, while BjRBRs, Retinoblastoma-related family genes regulate cell division and nuclear re-replication.

[0029] Before the invention, the inventors had never imagined that the SPL10c gene in tumorous stem mustard was related to the formation of tumor expansion. The inventors also promoted the expansion of tumorous stems of tumorous stem mustard and increased the yield of tumorous stem mustard by overexpressing the BjuSPL10c gene.

[0030] In order to achieve overexpression of the BjuSPL10c gene, the present invention also provides an overexpression vector of the BjuSPL10c gene, and the construction method thereof comprises the following steps:

[0031] A pair of primers was synthesized, and the primer sequences were shown in SEQ ID NO.3 and SEQ ID NO.4.

[0032] Total RNA was extracted from the tumors of wild-type Brassica juncea and reverse transcribed into cDNA.

[0033] The cDNA was used as a template and the primers were used for PCR amplification to obtain the target gene BjuSPL10c of the tumorous stem mustard.

[0034] The expression vector and the target gene of stem tumor mustard BjuSPL10c were sequentially digested, connected and transformed to obtain an overexpression vector.

[0035] The present invention is further described in detail below in conjunction with the examples, but the embodiments of the present invention are not limited thereto. Unless otherwise specified, the technical means used in the following examples are conventional means well known to those skilled in the art; the experimental methods used are all conventional methods and can be completed according to the described recombinant technology, see Molecular Cloning, Laboratory Manual, 2nd Edition, Cold Spring Harbor Laboratory Press, Cold Spring Harbor, New York; MaX et al, Arobust CRISPR / Cas9 system for convenient, high-efficiency multiplex genome editing in monocot and dicot plants. Molecular Plant. 2015, 8 (8): 1274-1284.; the materials, reagents, etc. used can all be obtained from commercial sources.

[0036] Example 1: Construction of the overexpression vector of B. juncea BjuSPL10c

[0037] The wild-type tumorous stem mustard tubers stored at -80°C were used as materials. The total RNA of the wild-type tumorous stem mustard tubers was extracted using the Tiangen plant total RNA extraction kit RNAprep pure Plant Kit according to the instructions. 2 μL of total RNA was taken and reverse transcribed to synthesize the first-strand cDNA according to the instructions of All-in-One 5×RT MasterMix. All-in-One 5×RT MasterMix was purchased from abm, product number G592.

[0038] The reverse transcription program was: 37°C, 15 min; 60°C, 10 min; 95°C, 3 min.

[0039] The first-strand cDNA synthesized by reverse transcription was used as a template to amplify the BjuSPL10c coding sequence using the high-fidelity enzyme MegaFiTM Fidelity2×PCRMasterMix. PCR amplification system: Primix 25μL, 10μM BjuSPL10c-F forward primer 2μL, 10μM BjuSPL10c-R reverse primer 2μL, cDNA template 2μL, sterile ddH 2 The forward primer sequence is shown in SEQ ID NO.3, and the reverse primer sequence is shown in SEQ ID NO.4.

[0040] SEQ ID NO. 3: BjuSPL10c-F, 5'-GAGAACACGGGGGACTCTAGAATGGACTGCAACATGGTATCTCCG-3'.

[0041] SEQ ID NO. 4: BjuSPL10c-R, 5'-GCCCTTGCTCACCATTCTAGATCATAAAAAATGAGTAGGGAAG-3'.

[0042] The obtained PCR amplification products were analyzed by agarose gel electrophoresis. Figure 1 As shown, under ultraviolet light irradiation, a specific amplified band can be observed at 1kb to 2kb, namely the BjuSPL10c target gene. The BjuSPL10c target gene was recovered using an agarose gel recovery kit D1200 purchased from Solarbio and then used for later use.

[0043] Figure 1 This is the electrophoresis diagram of the BjuSPL10c gene amplification and expression vector restriction enzyme digestion of the stem tumor mustard. M: Marker; Lane 1: electrophoresis detection results after BjuSPL10c target gene amplification; Lane 2: electrophoresis detection results of the expression vector p1300-GFP restriction enzyme digestion product.

[0044] The p1300-GFP vector was linearized using the restriction endonuclease Xba I, which was purchased from Thermo Scientific Fast Digest, catalog number 91252720. The BjuSPL10c target gene obtained after amplification and recovery was recombined with the linearized p1300-GFP vector using the recombinant ligase 2×Pro Ligation-Free Cloning Master Mix to obtain a ligation product, which was then transformed into Escherichia coli DH5α. The recombinant ligase 2×Pro Ligation-Free Cloning Master Mix was purchased from abm, catalog number E086-1. The Escherichia coli DH5α containing the ligation product was plated and cultured, and three positive clones were picked from the LB culture plate containing 50 mg / L karatomycin sulfate for colony PCR detection, and then the bacterial solution containing the target band obtained by amplification was sent to BGI for sequencing and screening to obtain the overexpression vector p1300-BjuSPL10c-GFP. The results showed that the full-length sequence of the stem tumor mustard BjuSPL10c gene is shown in SEQ ID NO.1, including the stop codon, which contains a total of 1125bp reading frame. According to SEQ ID NO.1, the amino acid sequence of the stem tumor mustard BjuSPL10c was obtained by translation with Primer premier software, and its amino acid sequence is shown in SEQ ID NO.2, which contains 375 amino acid residues, where * indicates a stop signal.

[0045] Example 2: Application of the BjuSPL10c gene overexpression vector of Brassica juncea

[0046] The above overexpression vector p1300-BjuSPL10c-GFP was transformed into Agrobacterium GV3101 competent cells. Adjust the concentration of Agrobacterium OD 600The cotyledons of the seedlings of the tumorous stem mustard were infected with 0.05 for 10 minutes; the cotyledons of the seedlings were then placed in a co-culture medium for dark culture for 2 days, the formula of the co-culture medium being: adding 2 mg of 6-benzylaminopurine, 0.15 mg of α-naphthaleneacetic acid and 8 g of agar per liter of MS culture medium; the co-cultured cotyledons of the seedlings were then transferred to a callus induction medium for further culture to induce callus tissue of the tumorous stem mustard, the formula of the callus induction medium being: adding 2 mg of 6-benzylaminopurine, 0.15 mg of α-naphthaleneacetic acid, 20 mg of hygromycin, 500 mg of timentin and 8 g of agar per liter of MS culture medium; the callus tissue was ... The adventitious buds were transferred to an adventitious bud differentiation medium, the formula of which was as follows: 2 mg of 6-benzylaminopurine, 0.15 mg of α-naphthylacetic acid, 20 mg of hygromycin, 500 mg of timentin and 8 g of agar were added to each liter of MS medium; when the adventitious buds grew to 3 cm, they were transferred to a rooting medium, the composition of which was as follows: 0.1 mg of α-naphthylacetic acid, 20 mg of hygromycin, 500 mg of timentin and 8 g of agar were added to each half liter of MS medium; the rooted strains were transferred to nutrient soil, placed in a light culture room for culture, and the stem tumor mustard plants overexpressing the BjuSPL10c gene were obtained.

[0047] gDNA of the stem tumorous mustard plants overexpressing the BjuSPL10c gene was extracted and PCR was performed. Figure 2 As shown, the detection primers are: p1300-GFP-F and p1300-GFP-R, and the primer sequences are shown in SEQ ID NO.5 and SEQ ID NO.6. Figure 2 In the middle, M: Marker; Lanes 1 to 3: 3 wild-type Brassica juncea plants; Lanes 4 to 13: 10 T. 0 Tumor mustard plants.

[0048] SEQ ID NO.5: p1300-GFP-F, 5'-GAATTCGCATGCCTGCAGGTC-3';

[0049] SEQ ID NO.6: p1300-GFP-R, 5'-GTGACAGATAGCTGGGCAATG-3'.

[0050] The T cells overexpressing the BjuSPL10c gene that could amplify the GFP nucleotide fragment obtained in the preliminary screening were 0After the three generations of self-pollination, RNA was extracted from the plants. The expression of BjuSPL10c mRNA in the three generations of self-pollination was detected by qRT-PCR. The transgenic plants of BjuSPL10c with significantly increased expression of BjuSPL10c mRNA were screened. The results of BjuSPL10 mRNA expression were shown in Figure 2. Figure 3 As shown, the transgenic plants of tumorous stem mustard BjuSPL10c obtained by screening were transplanted into the experimental field to observe the tumor phenotype. Figure 3 This is a graph showing the results of detecting the expression level of BjuSPL10 cmRNA in the BjuSPL10c overexpressing tumor mustard strains after three generations of self-pollination. WT is the wild-type control, and OE4, OE8, OE9, OE10, OE11, OE12 and OE13 are the BjuSPL10c overexpressing tumor mustard strains after three generations of self-pollination.

[0051] OE4, OE8, OE9, OE10, OE11, OE12, and OE13 were cultured, and the weight of the tubers was observed and measured on the 30th, 50th, 70th, 90th, 100th, and 130th days of their growth. Among them, OE12 had the highest expression of BjuSPL10c mRNA and the most significant phenotype. Only the phenotypic statistics of OE12 are shown here. The phenotypic statistical results are shown in Figure 4 Compared with the wild type, the tumor weight and volume of the tumorous stem of the B. juncea plants overexpressing BjuSPL10c were significantly increased. Figure 4 Figure 2 is a diagram of the enlarged tumor phenotype of the BjuSPL10c overexpressing stem tumor mustard strain and the wild-type plant. WT is the wild-type control, OE12 is the BjuSPL10c overexpressing strain 12, Figure 4 a is the statistics of tuber weight of BjuSPL10c overexpressing plants; Figure 4 b shows the observation of tumor expansion phenotype in plants overexpressing BjuSPL10c.

[0052] The technical features of the above-described embodiments may be arbitrarily combined. To make the description concise, not all possible combinations of the technical features in the above-described embodiments are described. However, as long as there is no contradiction in the combination of these technical features, they should be considered to be within the scope of this specification.

[0053] The above-mentioned embodiments only express several implementation methods of the present invention, and the description is relatively specific and detailed, but it cannot be understood as limiting the scope of the invention patent. It should be pointed out that for ordinary technicians in this field, several modifications and improvements can be made without departing from the concept of the present invention, which all belong to the protection scope of the present invention.

Claims

1. A BjuSPL10c gene for regulating tumor expansion, characterized in that: Its nucleotide sequence is shown in SEQ ID NO.1; The amino acid sequence is shown in SEQ ID NO.

2.

2. An overexpression vector constructed using the BjuSPL10c gene of claim 1.

3. The method for constructing an overexpression vector according to claim 2, characterized in that: The following steps are involved: Synthesize a pair of primers, the primer sequences are shown in SEQ ID NO.3 and SEQ ID NO.4; Total RNA was extracted from the tumor stems of wild-type Brassica juncea and reverse transcribed into cDNA; Using cDNA as a template and the primers to perform PCR amplification, the target gene of BjuSPL10c of Brassica juncea was obtained; The expression vector and the target gene of stem tumor mustard BjuSPL10c were sequentially digested, connected and transformed to obtain an overexpression vector.

4. The method for constructing an overexpression vector according to claim 3, characterized in that: The tuber is a tuber in the middle stage of swelling.

5. The method for constructing an overexpression vector according to claim 3, characterized in that: The expression vector is p1300-GFP.

6. Use of the tumorous stem mustard BjuSPL10c gene according to claim 1 or the overexpression vector according to claim 2 in promoting tumor expansion of tumorous stems of tumorous stem mustard.

7. The use according to claim 6, characterized in that: The overexpression vector is used to prepare a tumorous stem mustard plant that overexpresses the BjuSPL10c gene, and the preparation process includes the following steps: The overexpression vector is transformed into Agrobacterium competent cells to obtain positive recombinant Agrobacterium; The positive recombinant Agrobacterium is infecting the cotyledons of the tumorous mustard seedlings, and the cotyledons of the tumorous mustard seedlings are transferred to the co-culture medium and cultured in the dark; The cotyledons of the tumorous mustard seedlings were transferred in sequence to a callus induction medium to produce callus, an adventitious bud differentiation medium to produce adventitious buds, and a rooting medium to produce roots, thereby obtaining tumorous mustard plants overexpressing the BjuSPL10c gene.

8. The use according to claim 7, characterized in that: When the positive recombinant Agrobacterium infects the cotyledons of the tumorous mustard seedlings, the bacterial solution OD 600 is 0.

05.

9. The use of the overexpression vector according to claim 7, characterized in that: The callus induction medium formula is: Add 2 mg of 6-benzylaminopurine, 0.15 mg of α-naphthylacetic acid, 20 mg of hygromycin, 500 mg of timentin and 8 g of agar per liter of MS medium; The formula of adventitious bud differentiation medium is: Add 2 mg of 6-benzylaminopurine, 0.15 mg of α-naphthylacetic acid, 20 mg of hygromycin, 500 mg of timentin and 8 g of agar per liter of MS medium; The rooting medium formula is: 0.1 mg of α-naphthylacetic acid, 20 mg of hygromycin, 500 mg of timentin and 8 g of agar were added to each half liter of MS medium.

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