Application of tomato SlFMO1 gene in regulation and control of growth and development of tomato plants

Abstract

The invention discloses an SlFMO1 gene and application thereof in regulation and control of growth and development of tomato plants, and belongs to the technical field of gene engineering. According to the method, a CRISPR / Cas9 gene editing technology is utilized, firstly, a CRISPR / Cas9-gRNA vector is constructed to accurately edit an SlFMO1 gene in tomatoes, GV3101 agrobacterium tumefaciens is utilized to conduct mediated transformation on the tomatoes, and kanamycin serves as a resistance marker to conduct screening to obtain SlFMO1-deleted mutant tomatoes. Phenotype observation and analysis show that after the SlFMO1 gene is knocked out, the plant height and the plant width of a tomato plant are obviously increased, the apical development is enhanced, the stem diameter of the plant is increased, and the inflorescence spacing is increased, which indicates that the SlFMO1 gene negatively regulates the growth and development of the tomato, and meanwhile, the expression of related genes synthesized by auxin, gibberellin and brassinolide is obviously enhanced. The discovery has important theoretical significance and practical application value for follow-up cultivation of ideal tomato varieties.

Description

Technical Field

[0001] This invention belongs to the field of genetic engineering technology, specifically relating to tomatoes. SlFMO1 Application of genes in regulating the growth and development of tomato plants. Background Technology

[0002] Tomatoes are an important economic horticultural crop widely cultivated globally. Plant type characteristics, as key agronomic traits determining tomato yield, suitability for mechanized cultivation, and land use efficiency, have always been important targets for variety improvement. Among them, plant height is a core indicator of plant type, affecting not only ventilation and light penetration and lodging resistance, but also determining planting density per unit area, which is of great significance for modern greenhouse cultivation and urban agriculture. Generally, taller tomatoes have stronger light acquisition capabilities and larger fruiting space, providing a better biological basis for high yield potential. Therefore, identifying and regulating key genes that regulate tomato plant height has important theoretical and practical application value for cultivating ideal plant types and improving tomato yield efficiency.

[0003] Existing research on strategies for improving tomato plant height regulation mainly focuses on the intervention and application of genes related to endogenous plant hormones (such as gibberellins, auxins, and brassinolide). However, plant height regulation strategies based on single hormone pathways have certain limitations in practical applications. Global interference with hormone synthesis or signal transduction often leads to multiple negative effects. Therefore, overcoming the shortcomings of existing global hormone regulation and creating new tomato germplasm that balances ideal plant architecture and stable yield is a pressing technical challenge in the field of horticultural molecular breeding. Summary of the Invention

[0004] The technical problem to be solved by this invention is: to provide tomatoes SlFMO1 The application of genes in regulating the growth and development of tomato plants aims to solve the technical problem of synergistically improving ideal plant type and stable yield.

[0005] To achieve the above objectives, the technical solution adopted by the present invention is: to provide a method for regulating the growth and development of tomato plants. SlFMO1 Gene, SlFMO1 The nucleotide sequence of the gene is as follows: a. A nucleotide sequence as shown in SEQ ID NO: 1; or b. Under stringent conditions, the nucleotide sequence is identical to that shown in SEQ ID NO: 1, and encodes a functional protein that regulates the growth and development of tomato plants; or c. A nucleotide sequence as shown in SEQ ID NO: 1, wherein one or more nucleotides have been substituted, deleted, and / or added, and / or one or more base pairs have been mutated, and which encodes a functional protein that regulates the growth and development of tomato plants.

[0006] The present invention also discloses a method comprising the above. SlFMO1 Recombinant gene expression vectors.

[0007] The present invention also discloses an engineered bacterium comprising the above-mentioned recombinant expression vector.

[0008] The present invention also discloses the above-mentioned SlFMO1 Application of genes, recombinant expression vectors, or engineered bacteria in regulating the growth and development of tomato plants.

[0009] Based on the above technical solution, the present invention can be further improved as follows: Furthermore, by knocking out SlFMO1 Genes enable the regulation of tomato plant growth and development.

[0010] Furthermore, by knocking out SlFMO1 Genetic changes significantly increased tomato plant height and width, enhanced apical development, increased stem thickness, and wider inflorescence spacing; simultaneously SlTAR2 , SlToFZY4 , SlToFZY8 , SlGA20x1 , SlGA20x2 , SlBZR1 and SlCYPB3 Gene expression was significantly increased.

[0011] This invention also discloses a method for promoting the growth and development of tomato plants, comprising the above-mentioned... SlFMO1 Gene knockout expression vectors were transferred into Agrobacterium and then used to infect tomato explants. After cultivation, the height and width of tomato plants increased significantly, apical development was enhanced, stem diameter increased, and inflorescence spacing increased, thus promoting the growth and development of tomato plants.

[0012] The beneficial effects of this invention are as follows: This invention utilizes CRISPR / Cas9 gene editing technology, firstly constructing a CRISPR / Cas9-gRNA vector to precisely edit the SlFMO1 gene in tomato, and then using GV3101 Agrobacterium-mediated transformation of tomatoes, using hygromycin as a resistance marker to screen for SlFMO1-deficient mutant tomatoes. Phenotypic observation and analysis revealed that knockout… SlFMO1 Genetically modified tomato plants showed significantly increased plant height and width, enhanced apical development, larger stem diameter, and wider interflorescence spacing, indicating... SlFMO1 Genes negatively regulate the growth and development of tomatoes. Subsequent analysis of the expression of related genes affecting growth and development revealed... SlFMO1 Gene knockout lines of plants with auxin synthesis-related genes ( SlTAR2 , SlToFZY4 , SlToFZY8 ), gibberellin synthesis-related genes ( SlGA20x1 , SlGA20x2) and genes related to brassinolide synthesis ( SlBZR1 , SlCYPB3 The expression of ) was significantly increased, further validating the knockout effect. SlFMO1 Genes can promote the growth and development of tomato plants. This discovery has significant theoretical and practical value for subsequent improvements in tomato varieties and the cultivation of ideal tomato cultivars. Attached Figure Description

[0013] Figure 1 for SlFMO1 A schematic diagram of gene editing in gene knockout strains; Figure 2 Wild type and SlFMO1 Phenotypic diagram of plant height in gene knockout lines; Figure 3 Wild type and SlFMO1 Plant width phenotype of gene knockout lines; Figure 4 Wild type and SlFMO1 Statistical results of plant height of gene knockout lines; Figure 5 Wild type and SlFMO1 Statistical results of plant width of gene knockout lines; Figure 6 Wild type and SlFMO1 Statistical results of stem diameter of gene knockout lines; Figure 7 Wild type and SlFMO1 Statistical results of inflorescence spacing in gene knockout lines; Figure 8 Wild type and SlFMO1 In the leaves of gene knockout strains SlTAR2 Gene expression comparison; Figure 9 Wild type and SlFMO1 In the leaves of gene knockout strains SlToFZY4 Gene expression comparison; Figure 10 Wild type and SlFMO1 In the leaves of gene knockout strains SlToFZY8 Gene expression comparison; Figure 11 Wild type and SlFMO1 In the leaves of gene knockout strains SlGA20x1 Gene expression comparison; Figure 12 Wild type and SlFMO1 In the leaves of gene knockout strains SlGA20x2 Gene expression comparison; Figure 13 Wild type and SlFMO1 In the leaves of gene knockout strains SlBZR1 Gene expression comparison; Figure 14 Wild type and SlFMO1 In the leaves of gene knockout strains SlCYPB3 Gene expression comparison. Detailed Implementation

[0014] The specific embodiments of the present invention are described below to facilitate understanding of the invention by those skilled in the art. Unless otherwise specified, specific conditions are applied according to conventional conditions or the manufacturer's recommendations. Reagents or instruments whose manufacturers are not specified are all commercially available conventional products. However, it should be understood that the present invention is not limited to the scope of the specific embodiments. For those skilled in the art, various modifications are obvious as long as they fall within the spirit and scope of the invention as defined and determined by the appended claims. All inventions utilizing the concept of this invention are protected.

[0015] SlFMO1 The nucleotide sequence of the gene is as follows:

[0016] SlFMO1 The amino acid sequence of the protein encoded by the gene is as follows: MAIIRSKIGIIGGGISGIAAAKQLAKYDPMVFEATECVGGVWKHCSYRSTKLQTPRCDYEFSDFPWTQRDNTNFPTYEEILDYLYSYAKHFDVLKFFNFNSKVVEIKFVGDREMNYFGEYGSLLTGNPV WEVAVRNNQSQTLQWYAFDFVVMCTGKYGDIPNIPNFPPKKGPKEFNGQVLHTLDYSKLDQEASTKLMKGKKVVVGYKKSAIDLAVECAEANQGPEGQPCTMVVRTLHWTVPHYSIWGLPFYLFYSTR SSQFLHERPNQGLFRTLLCHMLSPMRKAASKLIESYLEWKLPLEKYGLKPEHPFEEDYASCQMAILPENFFTEADKGKIMFKRTSKWWFWEGGVEFEDNTKLEADVVILATGFDGKKKIKAILPDPFRS LVEFPSGMIPLYRGTIHPLIPNMAFVGYLESVSNLHSAEIRCIWLSRLVDDLFKLPSVEKMLEQITQEMEIMKRTTRFYKRNCISTFSINHSDEICQEMGWQVWRKTNWLAEAFSPYNSQDYAEEK (SEQ ID NO: 2).

[0017] The primer sequences used in the following examples are shown in Table 1.

[0018] Table 1 Primer sequence listing

[0019] Example 1 Construction SlFMO1 CRISPR / Cas9 gene knockout system 1. Target sequence selection: The target site was selected using the CRISPR / Cas9 target online design website (https: / / chopchop.cbu.uib.no). The selected target site sequence was T1 (nucleotides 98-118 of SEQ ID NO: 1). SlFMO1 Location in genes such as Figure 1 As shown.

[0020] 2. Based on the target site sequence of gRNA, design gRNA amplification primers gRNA-F (SEQ ID NO: 3) and gRNA-R (SEQ ID NO: 4).

[0021] 3. Construction of gRNA expression vector: (1) Using plasmids as amplification templates, PCR amplification was performed using gRNA-F and gRNA-R, followed by purification by agarose gel electrophoresis to obtain gRNA fragments. The PCR amplification system is shown in Table 2.

[0022] Table 2 PCR amplification system

[0023] The PCR amplification program was as follows: 98℃ for 3 min pre-denaturation; 98℃ for 15 s denaturation, 55℃ for 10 s annealing, 72℃ for 1 min extension, 35 cycles; store at 4℃.

[0024] PCR products were recovered using the Omega Gel Extraction Kit from Beijing Yuntai Biotechnology Co., Ltd.

[0025] 4. The above PCR product was ligated to a CRISPR vector using a cut-and-ligate method to obtain a2-hyg-gfp-nCas9-tomato U6- SlFMO1 Expression carriers. The connection system is shown in Table 3.

[0026] Table 3 PCR Ligation System

[0027] The reaction conditions were: 37℃ for 5 min, 16℃ for 5 min, 50 cycles; 65℃ for 5 min; and stored at 16℃.

[0028] Example 2 Obtained SlFMO1 Gene knockout line plants 1. Screening of engineered bacteria (1) Transformation of Escherichia coli Place a2-hyg-gfp-nCas9-tomato U6- SlFMO1 The expression vector was transformed into Escherichia coli DH5α and then plated on LB medium containing 50 mg / mL kanamycin. Single colonies were picked the next day for PCR amplification to check for successful ligation. The PCR primers were upstream primer F1 (SEQ ID NO: 5) and downstream primer R1 (SEQ ID NO: 6). The PCR amplification band was approximately 600 bp.

[0029] (2) Transformation of Agrobacterium Select a single colony that was correctly amplified by PCR, extract plasmids, and sequence them. The sequence of sequencing primer R is shown in SEQ ID NO: 7.

[0030] The plasmid with completely correct sequencing was transformed into Agrobacterium GV3101 and plated on LB solid medium containing 50 mg / mL rifampin and 50 mg / mL kanamycin. After a single colony grew, colony PCR was performed to verify whether it was a positive clone.

[0031] 2. Tomato conversion (1) Seed disinfection In a clean bench, prepare two 50mL test tubes, one with 75% alcohol and the other with 10-20% available chlorine sodium hypochlorite, to sterilize tomato seeds. First, soak the seeds in 75% alcohol for 2 minutes, then rinse three times with sterile water. Next, soak them in 10% (v / v) sodium hypochlorite solution for 10 minutes, then rinse six to seven times with sterile water. Inoculate the seeds into pre-prepared, sterilized 1 / 2 MS (Murashige and Skoog Medium) medium at room temperature, inoculating 25 seeds into each medium. Seal the medium and incubate in the dark. After 2-3 days of dark incubation at 25℃, transfer to light (16h light + 8h dark) for 5-7 days. Closely observe the cotyledon angle and true leaf growth on the 3rd-4th day under light.

[0032] (2) Cut cotyledons Add 0.2 mg / L of 2,4-D (2,4-Dichlorophenoxyacetic acid) and 0.1 mg / L of KT (Kinetin) to MS1 liquid medium (M519 medium). Remove seedlings with fully extended cotyledons, cut off the leaf blades, and then cut off a small cotyledon with a wound from each leaflet. Sow the cotyledons into MS1 medium, seal the seedlings, and culture them under light (16 h light + 8 h darkness) for 24-36 h.

[0033] (3) Infection Transformed Agrobacterium GV3101 was inoculated into LB medium containing 50 μg / mL rifampin (Rif) and 50 μg / mL kanamycin (Kan) and cultured at 28°C for 2 days. Single colonies were then picked and cultured in 2 mL of LB medium containing 50 μg / mL Rif and 50 μg / mL Kan in the dark at 28°C and 200 rpm for 1.5 days. Then, 100 μL of the bacterial culture was added to 10 mL of fresh LB medium containing 50 μg / mL Rif, 50 μg / mL Kan, and 100 μM acetylsyl syringone (AS) and cultured overnight at 28°C and 200 rpm. The cells were then collected by centrifugation at 4000 rpm for 10 min at room temperature, and centrifuged at 4000 rpm for 8 min in 2 mL of MS medium containing 1.0 mg / L indoleacetic acid (IAA) and 1.75 mg / L zeatin nucleoside (ZR). Finally, the bacterial cells were resuspended in 2 mL of MS salt medium, and the OD was adjusted. 600 Up to 0.3-0.4.

[0034] After suspending the bacterial culture, pour the culture into MS1 medium (MS liquid medium + 1.0 mg / L IAA + 1.75 mg / L ZR + 100 μM AS) containing 15 mL of infection solution, and adjust to OD200. 600 A suitable infection concentration is 0.3-0.4. Use tweezers to pick up explants from MS1 medium and place them in the infection solution with the wound side down. Wrap the medium in aluminum foil and place in a shaker at 28°C and 80 rpm for 10 minutes. Use sterilized tweezers to pick up the infected explants, place them on filter paper to absorb excess bacterial solution, and then place them on MS1 ​​medium. Incubate in the dark for 2 days.

[0035] (4) Transfer to MS2 callus culture After 48 hours of infection, the cotyledons were placed in MS2 medium (MS + 1.0 mg / L IAA + 1.75 mg / L ZR + 50 mg / L Kan + 200 mg / L Tim (Temperature)) and cultured at 26°C (16 hours light) / 18°C ​​(8 hours darkness) for 2 weeks.

[0036] (5) Transplantation of MS2 resistant buds Transfer the cotyledons that have grown a lot of callus to MS2 medium and culture for 1-2 weeks.

[0037] (6) Transplanting to MS3 for rooting culture The cotyledons of the resistant buds that have grown larger are transferred out, the callus is removed in a large dish, and then they are transferred to MS3 medium (MS + 0.2 mg / L IAA + 50 mg / L Kan + 200 mg / L Tim) and cultured for 2 weeks.

[0038] (7) Transplanting Transfer the mature and well-rooted seedlings out of the culture medium, add nutrient soil to sterilized pots, gently lift the plants from the MS3 medium, and wash the solid culture medium off the tomato roots in a slow stream of water to prevent fungal growth on the roots later. Dig a small hole in the center of the nutrient soil, gently place the tomato roots around the hole, press firmly, clearly label the variety type and insert it into the soil, cover each plant with a plastic cup, and place it in the culture room for cultivation.

[0039] 3. Identification of genetically modified plants Eight seedling lines were harvested through tomato transformation, denoted as CR- SlFMO1 -1 and CR- SlFMO1 -2. PCR was used to detect whether the target gene had been edited in the seedlings. Sequencing primers were designed based on the target site location, namely CR- SlFMO1 -F (SEQ ID NO: 8) and CR- SlFMO1 -R (SEQ ID NO: 9).

[0040] Genomic DNA was extracted from the leaves of eight transgenic lines and from the leaves of wild-type lines, and then PCR amplification was performed. The PCR reaction system is shown in Table 4.

[0041] Table 4. PCR reaction system for identifying transgenic edited plants

[0042] The PCR amplification program was as follows: 95℃ for 3 min pre-denaturation; 95℃ for 15 s denaturation, 55℃ for 10 s annealing, 72℃ for 1 min extension, 35 cycles; store at 4℃.

[0043] The PCR products were sequenced using CR- sequencing primers. SlFMO1 -F and CR- SlFMO1 -R. By comparing the sequencing results, plants that had undergone editing and those that had not were identified. The edited plants were determined to be either homozygous or heterozygous. If homozygous, phenotypic observation and recording were performed; if heterozygous, propagation was carried out to await the next generation of plants before sequencing. Seeds from the edited tomato T0 generation were collected, planted, and T1 generation seedlings were obtained for PCR testing. Further testing and screening of homozygous edited tomato plants followed.

[0044] like Figure 1 As shown, based on the sequencing results analysis, two homozygous lines were selected and denoted as CR- SlFMO1 -1 and CR- SlFMO1 -2, CR- SlFMO1 -1 represents the addition of a base G, CR- SlFMO1 -2 represents the addition of a base T.

[0045] Example 3 SlFMO1 Gene regulation of tomato plant growth and development 1. Comparison of growth phenotypes SlFMO1 Gene knockout line (CR-) SlFMO1 -1 and CR- SlFMO1 -2) and wild-type growing materials such as Figure 2 and Figure 3 As shown, it can be initially seen that SlFMO1 The plant height and width of the gene knockout line were significantly higher than those of the wild type, and they visually exhibited a distinctly tall phenotype.

[0046] right SlFMO1 The plant height, plant width, stem diameter, and distance between 2-3 inflorescences of the gene knockout line plants were statistically analyzed, and the results are as follows: Figures 4-7 As shown in Table 5.

[0047] Table 5. Statistical results of tomato plant height and inflorescence spacing.

[0048] As Figures 4-7 As shown in Table 5, compared to the wild type, SlFMO1 Gene knockout plants showed significantly increased plant height and width, larger stem diameter, and greater interflorescence spacing.

[0049] 2. Analysis of the expression of genes regulating tomato plant height Take wild type and SlFMO1 RNA was extracted from the young shoot tip tissue of gene knockout line plants, and RNA was used to analyze the positive transgenic plants (CR-). SlFMO1 -1 and CR- SlFMO1 -2) The expression levels of genes regulating tomato plant height were detected, including the expression of genes related to auxin, gibberellin, and brassinolide synthesis. The specific steps are as follows: (1) RNA extraction After grinding young shoot tip tissue with liquid nitrogen, 0.1 g of the sample was transferred to a 2.0 mL centrifuge tube frozen in liquid nitrogen. 1 mL of Trizol was added, and the tube was inverted and allowed to stand for 5 min. 200 μL of chloroform was added, and the tube was vigorously inverted for 15 s. The tube was then incubated at room temperature for 3 min and centrifuged at 12000 rpm for 15 min at 4 °C. 400 μL of the supernatant was transferred to a new 1.5 mL centrifuge tube, and an equal volume of isopropanol (stored at -20 °C) was added. The tube was gently inverted and mixed, then incubated at -20 °C for 10 min and centrifuged at 12000 rpm for 10 min at 4 °C. The supernatant was discarded, and 1 mL of 75% ethanol (diluted with DEPC water) was added to suspend the precipitate. The tube was centrifuged at 12000 rpm for 1 min, the supernatant was discarded, and the centrifuge tube was air-dried in a fume hood for 5-10 min. 40 μL of DEPC water was then added to dissolve the precipitate. RNA quality was detected by agarose gel electrophoresis, and RNA concentration was determined by absorbance method. The RNA was then stored at -80℃ for later use.

[0050] (2) RNA reverse transcription Based on the RNA sample concentration, the RNA samples were uniformly diluted to a specific concentration, and reverse transcription was performed using a reverse transcription kit (TakaraBio). The reverse transcription process was as follows: Prepare the following mixture in an RNase-free centrifuge tube: 5 μL of 4× All-in-one qRT SuperMix, 1 pg-1 μg of total RNA, and 20 μL of RNase-free ddH2O. Pipettes were used to mix the mixture, and after centrifugation, the tube was placed in a PCR instrument. The PCR instrument reaction program was set as follows: 50℃, 15 min; 85℃, 5 s. After the reaction program was completed, the product could be used directly for qPCR or stored at -20℃.

[0051] Determine auxin synthesis-related genes according to the following procedure and SYBR mix system. SlTAR2 , SlToFZY4 , SlToFZY8 ), gibberellin synthesis-related genes ( SlGA20x1 , SlGA20x2 ) and genes related to brassinolide synthesis ( SlBZR1 , SlCYPB3 The expression level of ) was determined, and the selected internal reference gene was Solyc11g005330. The reaction system is shown in Table 6.

[0052] Table 6 Reaction System Table

[0053] The primers for detecting the expression level of the internal reference gene Solyc11g005330 were QF (SEQ ID NO: 10) and QR (SEQ ID NO: 11).

[0054] Genes related to auxin synthesis That is, SlTAR2 The primers for gene expression detection are upstream primer F2 (SEQ ID NO: 12) and downstream primer R2 (SEQ ID NO: 13); SlToFZY4 The primers for gene expression detection are upstream primer F3 (SEQ ID NO: 14) and downstream primer R3 (SEQ ID NO: 15). SlToFZY8 The primers for gene expression detection were upstream primer F4 (SEQ ID NO: 16) and downstream primer R4 (SEQ ID NO: 17).

[0055] Gibberellin synthesis-related genes, namely SlGA20x1 The primers for gene expression detection are upstream primer F5 (SEQ ID NO: 18) and downstream primer R5 (SEQ ID NO: 19). SlGA20x2 The primers for gene expression detection were upstream primer F6 (SEQ ID NO: 20) and downstream primer R6 (SEQ ID NO: 21).

[0056] Genes related to brassinolide synthesis, namely SlBZR1 The primers for gene expression detection are upstream primer F7 (SEQ ID NO: 22) and downstream primer R7 (SEQ ID NO: 23); SlCYPB3 The primers for gene expression detection are upstream primer F8 (SEQ ID NO: 24) and downstream primer R8 (SEQ ID NO: 25).

[0057] The results of the detection of related gene expression levels are as follows: Figures 8-14 As shown, compared to the wild type (WT), SlFMO1 Gene knockout lines of plants with auxin synthesis-related genes ( SlTAR2 , SlToFZY4 , SlToFZY8 ), gibberellin synthesis-related genes ( SlGA20x1 , SlGA20x2 ) and genes related to brassinolide synthesis ( SlBZR1 , SlCYPB3 The expression of ) was significantly increased, further validating the knockout effect. SlFMO1 Genes can promote the growth and development of tomato plants.

Claims

1. Regulating the growth and development of tomato plants SlFMO1 Genes, characterized by, The SlFMO1 The nucleotide sequence of the gene is as follows: a. A nucleotide sequence as shown in SEQ ID NO: 1; or b. A nucleotide sequence that hybridizes under stringent conditions with the nucleotide sequence shown in SEQ ID NO: 1 and encodes a functional protein that regulates the growth and development of tomato plants; or c. A nucleotide sequence as shown in SEQ ID NO: 1, wherein one or more nucleotides have been substituted, deleted, and / or added, and / or one or more base pairs have been mutated, and which encodes a functional protein that regulates the growth and development of tomato plants.

2. A recombinant expression vector, characterized in that, Includes the claims 1 SlFMO1 Gene.

3. An engineered bacterium, characterized in that, It includes the recombinant expression vector as described in claim 2.

4. The claim 1 SlFMO1 The application of genes, the recombinant expression vector of claim 2, or the engineered bacteria of claim 3 in regulating the growth and development of tomato plants.

5. The application according to claim 4, characterized in that, By knocking SlFMO1 Genes enable the regulation of tomato plant growth and development.

6. The application according to claim 5, characterized in that, By knocking SlFMO1 Genetic changes significantly increased tomato plant height and width, enhanced apical development, increased stem thickness, and wider inflorescence spacing; simultaneously SlTAR2 , SlToFZY4 , SlToFZY8 , SlGA20x1 , SlGA20x2 , SlBZR1 and SlCYPB3 Gene expression was significantly increased.

7. A method for promoting the growth and development of tomato plants, characterized in that, Will include the contents of claim 1 SlFMO1 Gene knockout expression vectors were transferred into Agrobacterium and then used to infect tomato explants. After cultivation, the height and width of tomato plants increased significantly, apical development was enhanced, stem diameter increased, and inflorescence spacing increased, thus promoting the growth and development of tomato plants.

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