Application of gene GmSW19 in regulation and control of plant grain weight and quality

Abstract

The invention discloses an application of a gene GmSW19 in regulation and control of plant grain weight and quality. GWAS is developed by using genotype and hundred-grain weight phenotype data, haplotype analysis, gene expression profile data and homologous gene function annotation are combined, and the grain weight related gene GmSW19 is successfully cloned by virtue of research means of molecular biology and comparative genomics. The function of the gene GmSW19 in regulating and controlling the grain weight and quality of the plant is disclosed for the first time through a CRISPR-Cas9 system, the target of increasing the hundred-grain weight, fat and per unit yield of the plant is possibly achieved by knocking out a coding sequence of the gene GmSW19 or silencing the coding gene of the gene GmSW19 or reducing the expression level of the gene GmSW19, an important gene resource is provided for molecular breeding of the plant, and the application of the gene GmSW19 in regulating and controlling the grain weight and quality of the plant is also facilitated. And a new choice is provided for high yield of plants.

Description

Technical Field

[0001] This invention relates to the field of plant genetic engineering technology, and more specifically, to the application of the gene GmSW19 in regulating plant grain weight and quality. Background Technology

[0002] Soybeans Glycine max (L.) Merr. is a dual-purpose crop (food and forage) with significant strategic importance globally, and is also one of my country's main sources of plant protein and edible oil. However, my country's soybean industry is facing an extremely severe situation. my country's high dependence on soybean imports not only poses a serious threat to my country's food supply security but also carries potential risks to national economic security. Many factors constrain the growth of my country's soybean production, with limited planting area and low yield (only about two-thirds of the world average) being key bottlenecks. Given the current reality that it is difficult to significantly expand the planting area, increasing soybean yield has become the only way to enhance my country's soybean production capacity and meet industry demand.

[0003] 100-grain weight is a key factor determining soybean yield and is also closely related to soybean quality. However, although several genes related to 100-grain weight have been cloned, the genetic resources available for practical yield improvement remain extremely limited. Furthermore, most functional gene studies tend to focus on single traits, and a systematic understanding of the molecular mechanisms that simultaneously regulate grain weight and yield is still lacking. Therefore, it is necessary to identify soybean grain weight and quality-related genes through multi-omics association studies. This will not only help to recover "lost heritability" but also provide crucial genetic resources, molecular markers, and high-quality breeding materials for the creation of new high-yielding soybean germplasm through multi-gene aggregation, thereby driving breakthrough progress in soybean genetic improvement.

[0004] At present, the research on seed size regulation network mainly focuses on rice and Arabidopsis. These studies reveal that ubiquitin-proteasome pathway, mitogen-activated protein kinase (MAPK) signaling pathway, G protein signaling pathway, plant hormone signaling pathway and transcriptional regulatory factors play important roles in the regulation of seed size. Among them, bZIP transcription factors, as an important class of transcriptional regulatory factors, their protein structure is mainly composed of two parts: one is the basic amino acid region that specifically binds to DNA, and the other is the leucine zipper domain that participates in the formation of homodimer or heterodimer (Jakoby et al., 2002). Studies have shown that bZIP transcription factors are widely involved in multiple processes of plant growth and development, including seed germination and development (Wang et al., 2020; Correa et al., 2008), flower development (Muszynski et al., 2006) and so on. In addition, bZIP transcription factors also play a key role in plant resistance to stress, such as drought (Yang et al., 2020), salt stress (Gao et al., 2011), low temperature (Liao et al., 2008) and biological stress (He et al., 2020) and so on. However, there is no report on the bZIP family members in soybean that are involved in the regulation of seed size and quality at the same time. Therefore, identifying and further studying the bZIP family members that regulate soybean seed size and quality will provide new gene resources and solid theoretical support for improving soybean yield and quality, and thus effectively promote the process of soybean genetic improvement and promote the sustainable development of agricultural production. SUMMARY

[0005] The purpose of the present application is to provide the application of gene GmSW19 in regulating plant kernel weight and quality.

[0006] In order to achieve the purpose of the present application, in the first aspect, the present application provides the application of gene GmSW19 in regulating plant kernel weight and quality.

[0007] The gene GmSW19 of the present application is Glyma.19g194500 derived from soybean Willimas82, which is a gene encoding the following protein (a) or (b): (a) a protein consisting of the amino acid sequence shown in SEQ ID NO: 2; (b) a protein derived from (a) by substituting, deleting or adding one or more amino acids in the sequence shown in SEQ ID NO: 2 and having equivalent function.

[0008] The nucleotide sequence of gene GmSW19 is: i) the nucleotide sequence shown in SEQ ID NO: 1; ii) a nucleotide sequence of SEQ ID NO: 1 substituted, deleted and / or increased by one or more nucleotides and expressing the same functional protein; iii) a nucleotide sequence hybridizing to the sequence of SEQ ID NO: 1 under stringent conditions, which are hybridization in 0.1x SSPE or 0.1x SSC containing 0.1% SDS at 65°C, and washing the membrane with the same solution, and expressing the same functional protein; or iv) a nucleotide sequence having more than 90% homology to the nucleotide sequence of i), ii) or iii) and expressing the same functional protein.

[0009] Further, the gene GmSW19 negatively regulates the grain weight of a plant.

[0010] Further, the quality includes the fat content of a plant. The gene GmSW19 negatively regulates the fat content of a plant.

[0011] In the present application, the plant includes soybean.

[0012] In a second aspect, the present application provides a method for increasing the grain weight and improving the fat content of soybean, which comprises weakening or knocking out the gene GmSW19 in soybean by genetic engineering.

[0013] Further, the weakening includes silencing or reducing the expression level of the gene GmSW19 in soybean.

[0014] In a specific embodiment of the present application, the method comprises: taking the gene GmSW19 as a target, designing sgRNA sequence based on CRISPR-Cas9, connecting the DNA fragment containing the sgRNA sequence to the vector carrying CRISPR-Cas9, transforming soybean, and then obtaining the transgenic soybean with the gene function deficiency.

[0015] Preferably, the nucleotide sequence of the sgRNA action site is 5'-TGAACATGGACGAGTTCCTCA-3'.

[0016] In a third aspect, the present application provides the application of the transgenic soybean obtained by the method in plant breeding.

[0017] The breeding method includes but is not limited to transgenesis, crossbreeding, backcrossing, selfing or asexual reproduction.

[0018] By means of the above technical solution, the present application has at least the following advantages and beneficial effects: The application utilizes genotype and hundred-grain weight phenotype data to carry out GWAS, combines haplotype analysis, gene expression profile data and homologous gene function annotation, and successfully clones a grain weight related gene by means of molecular biology and comparative genomics research means GmSW19 . The gene is located in the reported QTL interval ( Figure 1 ).

[0019] The CRISPR-Cas9 gene editing system is used to edit the gene GmSW19 , and the results show that the hundred-grain weight of the mutant plant is significantly increased. It can be reasonably inferred that knocking out the coding sequence of the gene, or silencing the gene, or reducing the expression level of the gene, can achieve the goal of increasing the hundred-grain weight, fat and yield of plants. GmSW19

[0020] The application discloses the function of the gene GmSW19 in regulating grain weight of plants for the first time by the CRISPR-Cas9 system, which not only provides important gene resources for plant molecular breeding, but also provides a new choice for plant high yield. BRIEF DESCRIPTION OF DRAWINGS

[0021] Figure 1 The gene of the application is identified. Wherein, a: Manhadun diagram of hundred-grain weight trait; b: linkage disequilibrium LD diagram in the interval; c: expression heat map of the candidate gene; d: GmSW19 : tissue expression specificity expression analysis of the gene. GmSW19

[0022] The phenotype identification of the gene editing mutant in the preferred embodiment of the application is shown. Figure 2 GmSW19 The phenotype identification of the gene overexpression strain in the preferred embodiment of the application is shown.

[0023] Figure 3 The specific implementation GmSW19 The following examples are used to illustrate the application, but not to limit the scope of the application. If not specifically indicated, the technical means used in the examples are conventional means known to those skilled in the art, and the raw materials used are commercially available.

[0024] The following examples are used to illustrate the application, but not to limit the scope of the application. If not specifically indicated, the technical means used in the examples are conventional means known to those skilled in the art, and the raw materials used are commercially available.

[0025] ​​​The vector pCAMBIA3301 used in the following examples was purchased from Beijing Zhuangmeng International Biotechnology Co., Ltd., and the vector 0641 was provided by Mr. Liu Bin of the Institute of Crop Science, Chinese Academy of Agricultural Sciences (vector 0641, see the literature Lyu, X. et al. 2021, GmCRY1s modulate gibberellin metabolism to regulate soybean shade avoidance in response to reduced blue light. Mol. Plant, 14, 298-314.).

[0026] Example 1 Vector construction Construction of gene knockout vector: The sgRNA of the gene was designed using the software CRISPRP v2.0 (http: / / crispr.hzau.edu.cn / cgi-bin / CRISPR2 / CRISPR), and the 21bp sgRNA sequence (5'-TGAACATGGACGAGTTCCTCA-3') was located on the first exon of the gene. The U6 promoter sequence, sgRNA and zCas9 were sequentially connected using bridge PCR, and then constructed into the knockout vector pCAMBIA3301 using In-fusion. GmSW19

[0027] Construction of gene overexpression vector: The CDS sequence was cloned and inserted into the overexpression vector 0641. GmSW19

[0028] 1. Vector enzyme digestion 1) Reaction system: 10 × buffer 5 μL Bsa I 1 μL pCAMBIA3301 1 μg (20 μL) ddH2O 24 μL 10 × buffer 5 μL Xba I / Kpn I 1 μL / 1 μL 0641 1 μg (20 μL) ddH2O 23 μL 2) Reaction conditions: 37°C water bath for 1 h. Run the gel and recover using agarose gel electrophoresis.

[0029] 2. In-fusion ligation ​​The reagent used was Clontech 5 × In-Fusion® HD Enzyme Premix.

[0030] The recovered DNA fragment was ligated with the linearized carrier recovered after enzyme digestion.

[0031] 1) Reaction system: In-fusion 1 µL DNA fragment 2 µL Carrier fragment 2 µL 2) Reaction conditions: 50℃, 30 min.

[0032] 3, E. coli Trans10 transformation 1) Take Trans10 competent cells from the -80℃ refrigerator and place on ice for thawing, add all the ligation products to a 1.5 mL EP tube containing 50 µL of competent cells, mix gently and stand on ice for 30 min; 2) Heat shock at 42℃ water bath for 1 min, then quickly stand on ice for 2 min; 3) Add 500 µL of LB medium without antibiotics to each centrifuge tube, and place in a 37℃ shaker, 200 rpm, and shake for 1 h; 4) Take 100 µL of bacterial solution and evenly spread on LB plate medium containing kanamycin antibiotic, and incubate at 37℃ overnight.

[0033] 4, Identification of positive clones Use the gun head to pick single colonies in 10 µL of water, mix gently, and take 1 µL for colony PCR identification. The rest of the bacterial solution is added to LB liquid medium containing kanamycin antibiotic for expansion and culture for plasmid extraction.

[0034] 1) Reaction system: 2 × KOD mix 5 µL Primer (F / R) 0.5 µL / 0.5 µL Single colony bacterial solution 1 µL ddH2O 3 µL 2) Reaction procedure: Preheat 95℃ for 3 min; denaturation 95℃ for 5 s, annealing 60℃ for 5 s, extension 68℃ for 5 s (1 kb / 10 s), 35 cycles; final extension 68℃ for 5 min.

[0035] The PCR product was detected by electrophoresis, the target fragment was cloned, the corresponding expanded bacterial solution was sequenced, the positive clone was obtained, and the company's return plasmid was used for subsequent experiments.

[0036] Obtaining of transgenic soybean plants After the plasmid containing GmSW19 cDNA was constructed, the Agrobacterium was electroporated and then used to transform soybean variety Williams 82.

[0037] 1. Preparation and transformation of Agrobacterium competence 1) Preparation of Agrobacterium competence Single colonies of Agrobacterium K599 and EHA105 were picked and placed in 5 mL LB liquid medium containing the corresponding antibiotics, K599 resistance: 100 μg / mL streptomycin; EHA105 resistance: 100 μg / mL rifampicin (Rif). Incubate at 28°C overnight; inoculate 500 μL of overnight culture into 50 mL of LB liquid medium containing the same antibiotics, and incubate at 28°C until the OD 600 is about 0.8, centrifuge at 4000 rpm for 5 min, discard the supernatant. Add 30 mL of 10% pre-cooled glycerol to suspend the bacteria, centrifuge at 4000 rpm for 5 min, discard the supernatant, and repeat this step. After discarding the supernatant, add 3 mL of 10% glycerol to suspend the bacteria, and divide them into 1.5 mL sterile centrifuge tubes (200 μL per tube), and store them in a -80°C freezer.

[0038] 2) Agrobacterium transformation Thaw 200 μL of competent cells, add 4 μg of plasmid DNA and mix well, then place in an electroporation cup. Push the electroporation cup into the electroporator, press the pulse key for 5 s, then add 500 μL of YEB liquid medium to the electroporation cup, and transfer to a 1.5 mL centrifuge tube. Incubate at 28°C, 180 rpm for 2 h. In a clean bench, take out 200 μL of bacterial solution and evenly spread it on YEB solid medium containing kanamycin and rifampicin, and incubate at 28°C for 36 h.

[0039] Example 3: Detection of CRISPR vector editing efficiency in soybean hairy roots (1) Seed disinfection: Select soybean seeds that are intact, full, smooth, round, and free of spots. Sterilize the beans with 4 mL of concentrated hydrochloric acid and 100 mL of NaClO generated chlorine gas, and remove them after 16-18 hours. Blow off the chlorine gas in a clean bench for about 0.5-1 h.

[0040] (2) Seed germination: Place the beans evenly on the germination medium, 10 per dish. Incubate in a greenhouse for about three days until the radicles are about 2 cm long.

[0041] (3) Cutting bean and adjusting OD value of bacteria liquid: cut cotyledon, take the bean which germinates for 3-5 days, cut off from the hypocotyl at 0.3-0.5 cm, divide the cotyledon into two and cut open, remove the top bud. Take the shaken bacteria liquid (K599 Agrobacterium, OD value 0.5-0.7, generally shake for two days before cutting the bean, shake for one day in the evening the day before, and use in the morning of the day), centrifuge for 10 min at 4000 rpm, resuspend in liquid co-culture medium (3.21 g / L vitamin B5+0.59 g / L MES+20 g / L sucrose) to make the OD value of bacteria liquid 0.6, add the adjusted bacteria liquid into the cut bean, shake by hand every 10 min, and immerse for 30 min, and then take out and blow for about 10 min. 600 =0.6, cut the bean, add the adjusted bacteria liquid into the cut bean, shake by hand every 10 min, and immerse for 30 min, and then take out and blow for about 10 min.

[0042] (4) Co-culture: place the filter paper which has been sterilized on the co-culture medium, and then place the seed immersed with the bacteria liquid evenly in the medium (25 / plate), and cultivate for 3 days in dark at 28°C.

[0043] (5) Induction culture: after 3 days of cultivation, the embryo sprouts, wash with sterile water and liquid induction medium with hormones for 4-5 times, to ensure that the Agrobacterium is washed and dried with filter paper. Insert the embryo sprout into the solid induction medium with the top up, and place in the greenhouse for illumination culture. The root hair grows out after 10-14 days of culture in the greenhouse.

[0044] (6) Take the root hair of soybean transformed with different vectors, 2-3 roots / tube, generally take 3 repeats, extract DNA by CTAB method, and perform PCR detection and sequencing.

[0045] (7) Select the site which can be edited (base deletion or addition) for soybean transformation.

[0046] The used medium (L) is as follows: Co-culture medium: 2.15 g B5 powder+30 g sucrose+3.9 g MES+2 mL B5 organic+2 mg / L ZR+7.2 g agar Liquid induction medium with hormones: 3.1 g B5 powder+30 g sucrose+3.9 g MES+2 mL B5 organic+1.5 mg / L 6-BA+1 mL / L timentin+1.5 mL / L cefotaxime+0.8 mL / L glucose Solid induction medium: 3.1 g B5 powder+30 g sucrose+3.9 g MES+2 mL B5 organic+1.5 mg / L 6-BA+1 mL / L timentin+1.5 mL / L cefotaxime+0.8 mL / L glucose+7.5 g agar 3, Soybean transformation (1) Seed sterilization: sterilize the beans with chlorine gas generated by the reaction of concentrated hydrochloric acid and NaClO, and shake the bacteria.

[0047] (2) Seed germination: soybean seeds were placed in a culture dish, and sterile water was added to soak for 9-16 h. The seeds were just suspended, the beans were cut in half, and a part of the embryo tip was removed. A wound was made in the meristem zone of the bean, and it was soaked in sterile water. In the afternoon, the shaken bacterial solution was centrifuged (4000 rpm, 10 min), the bacteria were adjusted, and the OD value of the bacterial solution was 0.4-0.6. The sterile water in the beans was poured out, the adjusted bacterial solution was added, and the beans were shaken in a shaking bed for 30 min (28°C, 200 rpm). The beans were taken out and blown for about 10 min, and then placed on a co-culture medium and cultured in the dark for 3 days.

[0048] (3) Agrobacterium infection: use a scalpel to cut off 1 / 2 of the hypocotyl, then cut the hypocotyl longitudinally in the middle of the two cotyledons, and remove the apical bud and lateral buds. The cotyledon nodal explants were placed in resuspended Agrobacterium solution (OD=0.6), and then placed in a 25°C incubator for 0.5-4 h, shaking every 10-20 min to ensure full contact between the explants and the bacterial solution. After the infection was completed, the excess Agrobacterium solution was carefully poured out, the explants were taken out and blown for about 10 min, and then placed on a co-culture medium and cultured in the dark at 26°C for 3 days.

[0049] (4) Recovery culture: the growing hypocotyls were cut off, 3-4 mm were retained, the wounded side was upward, and the explants were inserted into the recovery medium at an angle of 10-12 explants per dish. The culture was carried out in a tissue culture room at 26°C with 16 h light / 8 h dark for 7 days.

[0050] (5) Bud induction: the growing hypocotyls were cut off, 3-4 mm were retained, and the explants were inoculated onto bud induction medium at 5 per dish. The culture was carried out at 24°C with 16 h light / 8 h dark for 21 days.

[0051] (6) Bud elongation: at the late stage of multiple bud induction, the contaminated individuals were eliminated, the cotyledon part of the explants was cut off, and a new horizontal cut was made at the base of the growth point. The cut was downward and inoculated into the elongation medium at 5 per dish. The culture was carried out at 24°C with 16 h light / 8 h dark for 21 days. If the explants had elongated, they could be directly subjected to root induction; if the explants still had elongation, new bud induction medium was replaced and the culture was continued for 21 days.

[0052] (7) Root induction: when the elongated bud was elongated to 3-4 cm, the elongated bud was separated from the multiple buds at the base of the elongated bud, and inoculated into the rooting medium for root induction. The rooting period was generally 2 weeks. After 2 weeks of rooting, the sealing film on the culture dish was removed, and the opening was used to harden the seedlings for 3-5 days.

[0053] (8) Transplanting: transplant into soil, each one seedling on the table bean variety, gene name, rooting date and soil culture date. Add the right amount of water, green manure and slow-release fertilizer, cover with a layer of film under light, adapt to strong light, and remove the film after 3 days.

[0054] The medium (L) used is as follows: Co-culture medium: 2.15 g B5 powder + 30 g sucrose + 3.9 g MES + 2 mL B5 organic + 2 mg / L ZR + 7.2 g agar Recovery medium: 3.1 g B5 powder + 30 g sucrose + 3.9 g MES + 2 mL B5 organic + 0.5 mg / L 6-BA + 1 mL / L tem + 1.5 mL / L cef + 7.5 g agar Bud induction medium: 3.1 g B5 powder + 30 g sucrose + 3.9 g MES + 2 mL B5 organic + 1.5 mg / L 6-BA + 1 mL / L tem + 1.5 mL / L cef + 0.8 mL / L glucose + 7.5 g agar Extension medium: 4.33 g MS powder + 30 g sucrose + 3.9 g MES + 2 mL B5 organic + 0.1 mL / L IAA and 0.5 mL / L GA3 + 0.6 mL / L glucose + 1 mL / L ZR + 7.5 g agar Rooting medium: 3.1 g B5 powder + 20 g sucrose + 0.6 g MES + 2 mL B5 organic + 1 mL / L tem + 1.5 mL / L cef + 1 mL / L IBA + 7.2 g agar Example 4 GmSW19 Identification of gene knockout mutants In order to determine the transgenic positive plants, a fresh leaf was taken, DNA was extracted, PCR amplification was carried out using target specific primers, and sequencing was carried out. At the same time, in order to confirm the overexpression positive strain, RNA was extracted from the leaves of the plants and reverse transcribed into cDNA, and qPCR detection was carried out using specific primers.

[0055] Example 5 Phenotypic identification of gene editing materials The number of transgenic samples used in this example is 50, which has statistical significance.

[0056] As shown in Figure 2 , GmSW19 After gene knockout, the phenotype of increased hundred-grain weight, increased fat content and less protein content appeared in the mutant plants. Among them sw19-1 、 sw19-2 、 sw19-3Homozygous mutants of -37, +1 and -5 bp frameshift mutation types, respectively. a-h are the statistical data of seed length (a), width (b), hundred-grain weight (c), protein content (d), fat content (e), plant height (f), grain weight per plant (g) and plot yield (h), respectively.

[0057] The above experimental results show that, GmSW19 negatively regulates soybean hundred-grain weight, fat content and yield, and positively regulates protein content.

[0058] Example 6: Phenotype identification of transgenic soybean The number of transgenic samples used in this example is at least 20, which has statistical significance.

[0059] As shown in Figure 3 , after overexpression of GmSW19 gene, the phenotype of reduced hundred-grain weight and fat content appeared in transgenic plants. Among them, OE-1, OE-2, OE-3 and OE-4 represent different overexpression transgenic lines. a is the expression level of GmSW19 in transgenic lines, and b-d are the statistical data of hundred-grain weight (b), fat content (c) and protein content (d) of overexpression transgenic lines.

[0060] The above experimental results show that, GmSW19 is a negative regulator of soybean hundred-grain weight and fat content.

[0061] The present application uses CRISPR-Cas9 gene editing system to edit GmSW19 gene, and the results show that the hundred-grain weight and fat content of mutant plants are significantly increased, and the protein content is reduced. It can be reasonably inferred that by knocking out the coding sequence of GmSW19 gene, or silencing the gene, or reducing the expression level, it is possible to achieve the goal of increasing plant hundred-grain weight, fat and yield per hectare.

[0062] Although the present application has been described in detail above with general description and specific embodiments, some modifications or improvements can be made on the basis of the present application, which is obvious to those skilled in the art. Therefore, these modifications or improvements made on the basis of not deviating from the spirit of the present application, all belong to the scope of the present application claimed.

[0063] Reference: 1、Correa, L., Riano-Pachon, D., Schrago, C., dos Santos, R., Mueller-Roeber, B., Vincentz, M. (2008). The role of bZIP transcription factors ingreen plant evolution: adaptive features emerging from four founder genes.PLOS ONE, 3, e2944. 2、Gao, S., Chen, M., Xu, Z., Zhao, C., Li, L., Xu, H., Tang, Y.,Zhao, X., Ma, Y. (2011). The soybean GmbZIP1 transcription factor enhancesmultiple abiotic stress tolerances in transgenic plants. PLANT MOL BIOL, 75,537-553. 3、He, Q., Cai, H., Bai, M., Zhang, M., Qin, Y. (2020). A soybean bZIPtranscription factor GmbZIP19 confers multiple biotic and abiotic stressresponses in plant. INT J MOL SCI, 21, 4701-4720. 4、Hu, Y., Liu, Y., Wei, JJ. Zhang WK, Chen SY, Zhang JS. (2023).Regulation of seed traits in soybean. aBIOTECH, 4, 372-385. 5、Jakoby, M., Weisshaar, B., Drge-Laser, W., Vicente-Carbajosa, J.,Tiedemann, J., Kroj, T., Parcy, F. (2002). bZIP transcription factorsin Arabidopsis. TRENDS PLANT SCI, 7, 106-111. 6、Liao, Y., Zou, H., Wei, W., Hao, Y., Tian, A., Huang, J., Liu, Y.,Zhang, J., Chen, S. (2008). Soybean GmbZIP44 , GmbZIP62 and GmbZIP78 genes functionas negative regulator of ABA signaling and confer salt and freezing tolerancein transgenic Arabidopsis . PLANTA, 228, 225-240. 7、Muszynski, M., Dam, T., Li, B., Shirbroun, D., Hou, Z., Bruggemann,E., Archibald, R., Ananiev, E., Danilevskaya. (2006). Delayed flowering1encodes a basic leucine zipper protein that mediates floral inductive signalsat the shoot apex in maize. PLANT PHYSIOL, 142, 1523-1536. 8、Wang, Y., Hou, Y., Qiu, J., Wang, H., Wang, S., Tang, L., Tong, X.,Zhang, J. (2020). Abscisic acid promotes jasmonic acid biosynthesis via a‘SAPK10-bZIP72-AOC’pathway to synergistically inhibit seed germination inrice (Oryza sativa). NEW PHYTOL, 228, 1336-1353. 9、Yang, Y., Yu, T., Ma, J., Chen, J., Xu, Z. (2020). The soybean bZIP transcription factor gene GmbZIP2 confers drought and salt resistances in transgenic plants. INT J MOL SCI, 21, 670-689.

Claims

1. Use of gene GmSW19 in regulating plant grain weight and quality. wherein Gene GmSW19 is a gene encoding the following protein (a) or (b): (a) a protein consisting of the amino acid sequence shown in SEQ ID NO: 2; (b) a protein derived from (a) by substitution, deletion or addition of one or several amino acids and having equivalent function.

2. Use according to claim 1, characterized in that, Gene GmSW19 negatively regulates plant grain weight.

3. Use according to claim 1, characterized in that, The quality includes the fat content of the plant.

4. Use according to claim 3, characterized in that, Gene GmSW19 negatively regulates the fat content of the plant.

5. The use according to any one of claims 1 to 4, characterized in that, The plant includes soybean.

6. A method for increasing the soybean kernel weight and improving the fat content, characterized by, The method includes weakening or knocking out gene GmSW19 in soybean by genetic engineering means. Gene GmSW19 is as described in claim 1. The weakening includes silencing or reducing the expression level of gene GmSW19 in soybean.

7. The method of claim 6, wherein, The method includes taking gene GmSW19 as a target, designing sgRNA sequence based on CRISPR-Cas9, connecting a DNA fragment containing the sgRNA sequence to a vector carrying CRISPR-Cas9, transforming soybean, and then obtaining transgenic soybean with the gene function deficiency.

8. The method of claim 7, wherein, The nucleotide sequence of the sgRNA action site is 5'-TGAACATGGACGAGTTCCTCA-3'.

9. Use of the transgenic soybean obtained by the method according to any one of claims 6-8 in plant breeding.

10. Use according to claim 9, characterized in that, The breeding method includes transgenesis, crossbreeding, backcrossing, selfing or asexual reproduction.

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