Soybean gmads119 gene and application thereof in plant flowering regulation

Abstract

The application discloses a soybean GmMADS119 gene and application thereof in plant flowering regulation, and belongs to the technical field of molecular biology. The nucleic acid sequence of the soybean GmMADS119 gene is shown as SEQ ID NO:1, and the amino acid sequence coded by the nucleic acid sequence is shown as SEQ ID NO:2. The soybean GmMADS119 gene is constructed into a plant expression vector, and is transformed into Arabidopsis thaliana and soybean respectively to obtain transgenic Arabidopsis thaliana and transgenic soybean. Research finds that the soybean GmMADS119 gene can promote Arabidopsis thaliana to bloom under long-day and short-day conditions; the soybean GmMADS119 gene can promote soybean to bloom under long-day and partial long-day conditions. Thus, the soybean GmMADS119 gene has important application prospects in plant flowering regulation.

Description

Technical Field

[0001] This invention belongs to the field of molecular biology technology, specifically relating to a soybean GmMADS119 gene and its application in the regulation of plant flowering period. Background Technology

[0002] Soybean (Glycine max) is a globally important economic crop and oilseed crop, and a core source of plant protein and oil in the human diet. Soybean yield and quality are regulated by various agronomic traits, among which flowering time is a key factor determining its adaptation to different latitudinal planting environments and optimizing the balance between vegetative and reproductive growth. Due to significant differences in environmental conditions such as photoperiod and temperature across different regions, the same soybean variety often exhibits late flowering and excessive vegetative growth in high-latitude, long-day regions, while tending to flower early and have stunted growth in low-latitude, short-day regions. Both lead to reduced pod setting rate and decreased 100-seed weight, severely restricting the planting range and yield stability of soybeans. Therefore, elucidating the molecular regulatory mechanism of soybean flowering time and identifying key genes that can precisely regulate flowering traits is a core requirement for breeding widely adaptable, high-yielding soybean varieties.

[0003] The regulation of flowering time in plants is a complex network involving multiple pathways working synergistically. In the model plant Arabidopsis thaliana, six major regulatory pathways have been identified: photoperiod, gibberellin, vernalization, temperature, age, and autonomous pathways. Each pathway integrates endogenous and environmental signals, ultimately acting on flowering integrators (such as FT and SOC1) to achieve flowering transition. Among them, the MADS-Box transcription factor family is a core member of the flowering regulatory network. Genes in this family bind to target gene promoters through conserved MADS domains, regulating the expression of key genes at various stages of flower development. In Arabidopsis thaliana, the MADS-Box family gene SHORT VEGETATIVE PHASE (AtSVP) has been confirmed as a flowering repressor. Its mutants exhibit early flowering phenotypes under both long-day and short-day conditions, and participate in temperature response and gibberellin signal integration through interactions with genes such as FLC. In addition, SVP homologous genes (SVP-Like genes) have also been found to participate in the regulation of flowering time or bud dormancy in species such as kiwifruit, alfalfa, and mango, indicating that SVP family genes are functionally conserved in plant flowering regulation.

[0004] However, as an ancient tetraploid crop, soybean has a large amount of gene redundancy in its genome, and its flowering regulation mechanism is more complex than that of model plants such as Arabidopsis thaliana. Currently, there are still significant technological gaps in the functional study of soybean SVP-Like genes. 1. Unclear gene function identification: Although the function of SVP genes in Arabidopsis and other crops has been identified, there are as many as 175 members in the MADS-Box gene family in soybean (Phytozome database). The expression patterns, regulatory targets and specific effects on flowering time of most SVP-Like genes have not yet been elucidated, and key genes that can be used for breeding applications cannot be identified. 2. The photoperiod response mechanism remains to be elucidated: Soybean is a typical short-day crop, but the differences in the response of its SVP-Like genes to different photoperiods (long-day / short-day), and whether it affects flowering by regulating the FT family (such as GmFT2a, GmFT5a, the core soybean florigen genes), have not been clearly reported. 3. Untapped potential for gene application: In current soybean breeding, the regulation of flowering time mainly relies on traditional hybridization or chemical regulation (such as gibberellin treatment). The former has a long cycle (requiring 5-8 generations of screening) and poor directionality, while the latter is prone to pesticide residues and abnormal plant type. Molecular design breeding based on key flowering regulatory genes has not yet been applied on a large scale due to the lack of clear functional gene targets. 4. Gene redundancy leads to difficulties in functional verification: The ancient tetraploid nature of soybeans often results in functional redundancy between adjacent homologous genes. It is difficult to observe obvious phenotypes when a single gene is knocked out, which further hinders the functional analysis and application of soybean SVP-Like genes.

[0005] In summary, current research on soybean flowering time regulation lacks functionally validated key SVP-Like genes that respond to photoperiod signals and can directionally regulate flowering traits. This makes it difficult for molecular breeding to precisely improve the relationship between soybean flowering adaptability and yield. Therefore, identifying key SVP-Like genes in soybean that regulate flowering time and clarifying their functions and regulatory mechanisms is of great significance for overcoming current soybean breeding technology bottlenecks and developing widely adaptable and high-yielding varieties. Summary of the Invention

[0006] To address the problems existing in the prior art, the purpose of this invention is to provide the soybean GmMADS119 gene and its application in the regulation of plant flowering period.

[0007] To achieve the above objectives, the present invention adopts the following technical solution: The soybean GmMADS119 gene is characterized in that the amino acid sequence encoded by the gene is shown in SEQ ID NO:2.

[0008] Based on the above scheme, the nucleic acid sequence of the gene is shown in SEQ ID NO:1.

[0009] The application of the soybean GmMADS119 gene in the regulation of plant flowering period.

[0010] Based on the above scheme, the soybean GmMADS119 gene was constructed into an expression vector, transformed into plants, and its expression level in plants was increased, thereby promoting plant flowering.

[0011] Based on the above scheme, the method of transforming plants is one of Agrobacterium-mediated transformation, gene gun transformation, electroporation transformation, PEG transformation, and liposome transformation.

[0012] Based on the above scheme, the plant is soybean or Arabidopsis thaliana.

[0013] A method for promoting flowering in Arabidopsis thaliana involves constructing the coding region sequence of the soybean GmMADS119 gene into a plant expression vector, transforming it into Arabidopsis thaliana, and expressing it in Arabidopsis thaliana to promote flowering in Arabidopsis thaliana under long-day and short-day conditions; the nucleic acid sequence of the soybean GmMADS119 gene is shown in SEQ ID NO:1.

[0014] Based on the above scheme, the long-day conditions are: 16 hours of light at 23°C and 8 hours of darkness at 20°C; humidity of 55%; light exposure begins at 7:00 AM; and light intensity of 14000 Lx. The short-day conditions are: 10 hours of light at 23°C and 14 hours of darkness at 20°C; humidity of 55%; light exposure begins at 7:00 AM; and light intensity of 14000 Lx.

[0015] A method for promoting soybean flowering involves constructing the coding region sequence of the soybean GmMADS119 gene into a plant expression vector, transforming soybeans, and expressing the gene in soybeans to promote flowering of soybeans under long-day and slightly longer-day conditions; the nucleic acid sequence of the soybean GmMADS119 gene is shown in SEQ ID NO:1.

[0016] Based on the above scheme, the long-day conditions are 16 hours of light at 25°C and 8 hours of darkness at 23°C; the start time of light exposure is 8:00 AM; and the light intensity is 14000 Lx. The conditions for the longer daylight hours are: summer outdoor natural temperature, 14.5 hours of sunshine, and 9.5 hours of darkness.

[0017] Advantages of the technical solution of this invention This invention discloses a soybean GmMADS119 gene and its application in plant flowering regulation. The gene was constructed into a plant expression vector and transformed into Arabidopsis thaliana and soybean, respectively, to obtain transgenic Arabidopsis thaliana and transgenic soybean. The flowering of transgenic Arabidopsis thaliana under long / short day conditions was studied, and the results showed that the soybean GmMADS119 gene can promote flowering in Arabidopsis thaliana. The flowering of transgenic soybean under long / short day conditions was also studied, and the results showed that under long-day and slightly longer-day conditions, the soybean GmMADS119 gene can promote flowering in soybean, while under short-day conditions, it has no effect on flowering. Therefore, the soybean GmMADS119 gene has important application prospects in plant flowering regulation. Attached Figure Description

[0018] Figure 1 Analysis of GmMADS119 expression levels in different Arabidopsis thaliana lines; Figure 2 Phenotypic observation and statistical data of GmMADS119 transgenic Arabidopsis thaliana under long-day conditions (A and B are phenotypic observations of three Arabidopsis thaliana lines; C is the number of rosette leaves of three Arabidopsis thaliana lines). Figure 3 Phenotypic observations and statistical data of GmMADS119 transgenic Arabidopsis thaliana under short-day conditions (A and B represent phenotypic observations of three Arabidopsis thaliana lines; C represents the number of rosette leaves in the three Arabidopsis thaliana lines). Figure 4 Observation and statistical data of flowering phenotypes in soybean under long-day conditions (A shows the flowering phenotypes of transgenic soybean plants and Williams 82 plants, where the first, second, and third from the right are transgenic soybean plants, and the first from the left is a Williams 82 plant; B shows the number of flowering days for GmMADS119 transgenic soybean plants and Williams 82 plants; C shows the number of nodes on the main stem of GmMADS119 transgenic soybean plants and Williams 82 plants; *P<0.05, ***P<0.001, Bar=4cm). Figure 5 Detection of endogenous genes in soybean leaves under long-day conditions (A: GmMADS119 detection; B: GmFT2a detection; C: GmFT5a detection; D: AP1a detection). Figure 6Observation and statistical data of flowering phenotypes in soybean under short-day conditions (A shows the flowering phenotypes of transgenic soybean plants and Williams 82 plants, where the first, second, and third from the right are transgenic soybean plants, and the first from the left is a Williams 82 plant; B shows the number of flowering days for GmMADS119 transgenic soybean plants and Williams 82 plants; C shows the number of nodes on the main stem of GmMADS119 transgenic soybean plants and Williams 82 plants; *P<0.05, ***P<0.001, Bar=4cm). Figure 7 Detection of endogenous genes in soybean leaves under short-day conditions (A: GmMADS119 detection; B: GmFT2a detection; C: GmFT5a detection; D: AP1a detection). Figure 8 Phenotypic observation and data statistics of GmMADS119 transgenic and Williams 82 soybeans under natural light (longer day) conditions (A: flowering phenotype of transgenic soybean plants and Williams 82 plants, where the first, second, and third from the right are transgenic soybean plants, and the first from the left is a Williams 82 plant, Bar=4cm; B: plant height statistics; C: main stem node statistics; D: total number of pods per plant statistics; E: branch number statistics; *: P<0.05, **: P<0.01, ***: P<0.001, ****: P<0.0001). Figure 9 Phenotypic diagrams of GmMADS119 transgenic soybean and Williams 82 soybean seeds under natural light (longer day) conditions (A: 100-seed weight of GmMADS119 transgenic soybean and Williams 82 soybean; B: seed length of GmMADS119 transgenic soybean and Williams 82 soybean; C: seed width of GmMADS119 transgenic soybean and Williams 82 soybean; D: seed height of GmMADS119 transgenic soybean and Williams 82 soybean; Bar = 2cm). Detailed Implementation

[0019] The terminology used in this invention, unless otherwise specified, generally has the meanings commonly understood by those skilled in the art. The invention is further described in detail below with reference to specific embodiments and data. The following embodiments are merely illustrative and are not intended to limit the scope of the invention in any way.

[0020] Unless otherwise specified, the experimental methods used in the following embodiments are conventional methods, performed according to the techniques or conditions described in the literature in this field or according to the product instructions. Unless otherwise specified, the experimental materials, reagents, and chemicals used in the following embodiments can be obtained through general channels.

[0021] In the following embodiments, Soybeans (Williams 82) were provided by the Crop Applied Functional Genomics Laboratory of the College of Life Sciences, Qingdao Agricultural University; The Columbia-type Arabidopsis thaliana (Columbia-0) was provided by the Crop Applied Functional Genomics Laboratory of the College of Life Sciences, Qingdao Agricultural University.

[0022] The overexpression vectors pPTN1171 and DH5α were provided by the Crop Applied Functional Genomics Laboratory of the College of Life Sciences, Qingdao Agricultural University.

[0023] Example 1

[0024] Cloning of the GmMADS119 gene First, RNA was extracted from soybean Wi82 leaves and then reverse transcribed into cDNA. Using this as a template, PCR amplification was performed using the GmMADS119-F / R primer pair to obtain the full-length cDNA sequence of the GmMADS119 gene, as shown in SEQ ID NO:1. This gene coding sequence contains 705 nucleotides, encoding 234 amino acids, as shown in SEQ ID NO:2. The GmMADS119-F / R primer pair sequences are shown in SEQ ID NO:3 and SEQ ID NO:4.

[0025] The PCR reaction system was as follows: 10 µL of 2xKeyPo Master Mix, 1 µL of GmMADS119-F, 1 µL of GmMADS119-R, 1 µL of template, and ddH2O added to a total volume of 20 µL.

[0026] The PCR reaction procedure was as follows: 98℃ for 10s, 60℃ for 5s, 72℃ for 10s, 30 cycles; stored at 4℃.

[0027] SEQ ID NO:1 (5'→3'): ATGGTGAGAAGGAAGATACCATCAAGAAGATCGACGACGTAACTGCGAGGCAGGTGACATTCTCAAAGAGGAAGAGTGGTCTTTTCAAGAAGGCACGTGAGCTCTCTCTTCTCTGTGATGCTGAGATAGCACTCATAGTCTTTTCACCTGGTGGCAAGCTCTTCGACTATGGTAGTTCAAGTATGCAGAAGGTAATTGAAAGGCATATTTTACGGTCAGAGCTTAACCTTGAAAAATTAGACCAGTCGTGTCCTACGGAGCAGGTGAGATGTAATTATGCTGATTTGAAC AAGGAATTTGCAGACAGGACTCGTGAAATGAGACAACTGAATGGAGAAGAACTGCAAGGATTGACCCTAAGGGAACTTCAGAAATTAGAAGAACGACTTGACTCATCCTTGAACCGTGTTTATAAAGCGAAGGTTGAAAATTTTA TAAAAGAGATCGGTATTCTTAAGGAAAAGATGATAAAATTACCGAGGAATGAAATATGTTCGGTCCAGAGACATGAGCATGAACAAGGCCAGCTCTTTGACACCTCCCTCACGTTGGGGCTACCTTTTCCTGCTGGCTCAAAGTAG SEQ ID NO:2: MWHIHVFPHTVFSLESKDWARVAFASAFSIAKESSAFHRNKKMVRRKIPIKKIDDVTARQVTFSKRKSGLFKKARELSLLCDAEIALIVFSPGGKLFDYGSSSMQKVIERHILRSELNLEKLD QSCPTEQVRCNYADLNKEFADRTREMRQLNGEELQGLTLRELQKLEERLDSSLNRVYKAKVENFIKEIGILKEKGKKLMEDNMLIKQMIKLPRNEICSVQRHEHEQGQLFDTSLTLGLPFPAGS GmMADS119-F:5'-CCGCTCGAGATGGTGAGAAGGAAGATACC-3'(SEQ ID NO:3); GmMADS119-R: 5'-GCTCTAGACTACTTTGAGCCAGCAGGAA-3' (SEQ ID NO: 4).

[0028] Example 2

[0029] Obtaining the overexpression vector of GmMADS119 and the recombinant strain Xho I and Xba I restriction endonucleases were used to double-digest the full-length cDNA fragment of GmMADS119 obtained by PCR in Example 1 above and the overexpression vector pPTN1171 to ensure ligation between the fragment and the vector. The digested system was placed in a PCR instrument at 37°C for 1 hour. After digestion, the fragment and vector were recovered using the SteadyPure DNA Gel Extraction Kit from Eric Biotechnology. The recovered fragment and vector were ligated using the T4 DNA ligation method. After constructing the system, it was incubated overnight at 16°C to obtain the overexpression vector (GmMADS119-pPTN1171). Subsequently, the constructed overexpression vector was transformed into DH5α by heat shock, cultured overnight at 37°C, and single clones were picked and sent to the company for sequencing to obtain the correct recombinant bacteria.

[0030] Example 3

[0031] Obtaining GmMADS119 transgenic Arabidopsis thaliana 1) The *E. coli* plasmid containing the correct overexpression vector from Example 2 was extracted and transformed into *Agrobacterium* GV3101 (purchased from Shanghai Weidi Biotechnology Co., Ltd.) using a freeze-thaw method. *Agrobacterium*, which had been verified to be correct, was used to infect *Arabidopsis thaliana* (Columbia-0) plants to obtain transgenic *Arabidopsis thaliana* plants. Seeds from the T0 generation were harvested after maturity. The T0 generation *Arabidopsis thaliana* seeds were screened using MS medium supplemented with PPT (glyphosate, 5 mg / L). The screened seeds were vernalized at 4°C for 3 days and then cultured under light in an incubator. After approximately two weeks of culture, the *Arabidopsis thaliana* seedlings were transferred to potting soil for further cultivation. Care was taken not to break the roots of the seedlings during transplanting. Newly transplanted seedlings were covered with plastic wrap until they reached a certain size, at which point the plastic wrap was removed.

[0032] 2) qRT-PCR detection of gene expression levels Because false positives may occur in the Arabidopsis thaliana plants screened above, it is necessary to perform DNA-level positive seedling testing on the screened plants to prove that the obtained plants are successfully transformed positive plants. The specific operation is as follows: The expression level of the target gene GmMADS119 in transgenic Arabidopsis thaliana lines was analyzed by qRT-PCR. RNA samples were taken from the leaves of selected plants 2-3 weeks after plant growth. Three plants from each line were selected, and samples were taken from each plant, ensuring minimal differences in growth status among the three plants. A control group was obtained from Columbia-type Arabidopsis thaliana (Columbia-0). All samples were immediately stored in liquid nitrogen, followed by RNA extraction. The expression level of the RNA samples was then analyzed. 260 and A 280 The concentration and purity of the cDNA were recorded, and 1 μg of RNA was used for reverse transcription. Using the cDNA obtained from reverse transcription as a template, the relative expression level of the GmMADS119 gene was detected by qRT-PCR with specific primers.

[0033] The specific primer sequences for qRT-PCR to detect GmMADS119 gene expression and the primer sequences for the internal control Actin gene are as follows: GmMADS119-QRT-F1: 5'-GGTGGCAAGCTCTTCGACTA-3' (SEQ ID NO: 5); GmMADS119-QRT-R1: 5'-CATCTCACCTGCTCCCGTAGG-3' (SEQ ID NO: 6); Atactin-F: 5'-GCTCCTCTTAACCCAAAGGC-3' (SEQ ID NO: 7); Atactin-R: 5'-CACACCATCACCAGAATCCAGG-3' (SEQ ID NO: 8).

[0034] qRT-PCR reaction system: 10µL 2X SYBR Green Pro Taq HS Premix, 1µL cDNA, 0.4µL upstream primer, 0.4µL downstream primer, 0.4µL ROX Reference Dye, and ddH2O added to a total volume of 20µL.

[0035] The qRT-PCR reaction program was as follows: pre-denaturation at 95℃ for 30s; denaturation at 95℃ for 5s; annealing and extension at 60℃ for 30s, for a total of 40 cycles.

[0036] Add each component of the system to a 96-well plate, centrifuge to remove air bubbles, place the 96-well sample plate in an ABI PCR instrument for amplification, analyze the melting curve and CT value of the obtained qRT-PCR, and calculate the relative expression level.

[0037] The results are as follows Figure 1As shown, the GmMADS119 gene was highly expressed in the three GmMADS119 transgenic Arabidopsis lines (GmMADS119-2, GmMADS119-3, and GmMADS119-8), while no GmMADS119 gene expression was detected in the Columbia-type Arabidopsis (Columbia-0). Figure 1 ).

[0038] Example 4

[0039] Regulation of flowering in Arabidopsis thaliana by the GmMADS119 gene Three transgenic Arabidopsis thaliana lines (GmMADS119) and the Columbia-type Arabidopsis thaliana (Columbia-0) were planted under long-day and short-day conditions for phenotypic observation.

[0040] Under long-day conditions (16 hours of light at 23°C and 8 hours of darkness at 20°C), the GmMADS119 transgenic Arabidopsis lines (such as GmMADS119-2, GmMADS119-3, and GmMADS119-8) exhibited a significantly earlier flowering phenotype, bolting earlier, and a significantly fewer rosette leaves than the wild type. Figure 2 ).

[0041] Under short-day conditions (10h light / 23℃, 14h darkness / 20℃), the GmMADS119 transgenic Arabidopsis line still exhibited an early-flowering phenotype, but the number of rosette leaves was reduced compared to the wild type at the same time. Figure 3 ).

[0042] Example 5

[0043] Obtaining GmMADS119 genetically modified soybeans The E. coli plasmid containing the correct overexpression vector was extracted from Example 2 and transformed into Agrobacterium LBA4404 (purchased from Shanghai Weidi Biotechnology Co., Ltd.) by freeze-thaw method. The verified Agrobacterium was used to transform the Williams82 soybean variety. Regenerated shoots slowly grew from the cotyledon nodes of the soybean through infection. Transgenic soybean plants of positive regeneration T0 generation were obtained through tissue culture and screening.

[0044] Example 6

[0045] The regulatory role of the GmMADS119 gene in soybean flowering The three GmMADS119 transgenic soybean lines (GmMADS119-2, GmMADS119-5, GmMADS119-20) obtained in Example 5 and soybean Williams82 were planted under long-day and short-day conditions for phenotypic observation.

[0046] Observation of flowering time of transgenic soybeans under long-day conditions Under long-day conditions (16 hours of light, 8 hours of darkness, 25°C during light, 23°C during darkness, 8:00 AM light start, and 14000 Lx light intensity), the flowering time of three lines of the GmMADS119 transgenic soybean was observed to be earlier than that of Williams82. The results indicate that the GmMADS119 gene can promote soybean flowering under long-day conditions. Figure 4 This causes soybeans to flower earlier.

[0047] qRT-PCR detection of endogenous gene expression levels in leaves of transgenic soybean plants under long-day conditions After identifying positive GmMADS119 transgenic soybean seedlings by PCR, the relative expression levels of GmMADS119, GmFT2a, GmFT5a, and GmAP1a in the transgenic lines and Williams82 were detected using real-time quantitative PCR. Samples were taken 21 days after plant growth. Soybean leaves were cut off with scissors and placed in aluminum foil bags, then immediately immersed in liquid nitrogen. RNA was extracted from the samples and reverse transcribed into cDNA. Using cDNA as a template, the relative expression levels of each gene were detected by qRT-PCR.

[0048] The primers required for qRT-PCR are shown in SEQ ID NO:5~6 and SEQ ID NO:9~16, containing 3 biological replicates and 2 technical replicates. Here, the soybean housekeeping gene GmCons4 is selected as the internal reference gene, and the Williams82 plant is used as the control.

[0049] The specific primer sequences are as follows: GmFT2a-QRT-F: 5'-GGATTGCCAGTTGCTGCTGT-3' (SEQ ID NO: 9); GmFT2a-QRT-R: 5'-GAGTGTGGGAGATTGCCAAT-3' (SEQ ID NO: 10); GmFT5a-QRT-F: 5'-GCCTTACTCCAGCTTATACT-3' (SEQ ID NO: 11); GmFT5a-QRT-R: 5'-GGCATGCTCTAGCATTGCAA-3' (SEQ ID NO: 12); GmAP1a-QRT-F: 5'-GTGAACATGGGTGGCAATTAC-3' (SEQ ID NO: 13); GmAP1a-QRT-R: 5'-TGTCAAATGCCATACCAAAG-3' (SEQ ID NO: 14); GmCons4-RT-F: 5'-CGGTGGTTTCTATCTTGGCATC-3' (SEQ ID NO: 15); GmCons4-RT-R: 5'-GTCTTTCGCTTCAATAACCCTA-3' (SEQ ID NO: 16); The obtained qRT-PCR data were analyzed, and the relative expression levels of endogenous genes in different plants were calculated. The results showed that under long-day conditions, the relative expression levels of GmMADS119, GmFT2a, GmFT5a, and GmAP1a were significantly increased in the GmMADS119 transgenic soybean line. Figure 5 ).

[0050] Observation of flowering time of transgenic soybeans under short-day conditions Under short-day conditions (10 hours of light, 14 hours of darkness, temperature set to 25℃ regardless of light exposure, and light intensity set to 14000 Lx), plants began to flower 25 days after planting. The number of flowering plants was recorded daily, and flowering time was determined by the appearance of white flower buds. Observational results showed that the average flowering days of the GmMADS119 transgenic soybean line and the cultivated Williams82 were around 30 days. The GmMADS119 gene had no effect on the flowering time of the Williams82 soybean under short-day conditions. Figure 6 ).

[0051] qRT-PCR detection of endogenous gene expression levels in leaves of transgenic soybean plants under short-day conditions After identifying positive seedlings by PCR, the relative expression levels of GmMADS119, GmFT2a, GmFT5a, and GmAP1a in GmMADS119 transgenic soybean lines and Williams82 were detected using real-time quantitative PCR. The method was the same as above. The results showed that under short-day conditions, the relative expression levels of GmMADS119 and GmAP1a significantly increased in GmMADS119 transgenic soybean lines. Figure 7 ).

[0052] Observation of flowering time of transgenic soybeans under natural light (longer daylight) conditions Preliminary phenotypic observations were conducted on T1 generation transgenic soybean plants planted in the experimental field at the end of May. At that time, the planting area had approximately 14.5 hours of sunshine and 9.5 hours of darkness, which falls under conditions of relatively long daylight hours with natural light. Phenotypic observations of the GmMADS119 transgenic soybeans grown under natural light conditions in the T1 generation revealed that after maturity, the GmMADS119 transgenic soybean lines exhibited significantly reduced plant height, fewer main stem nodes, fewer effective branches, and significantly fewer total pods per plant compared to Williams82. Figure 8 The 100-seed weight decreased significantly, and the length, width, and height of the seeds were all smaller than those of Williams82. Figure 9 ).

[0053] The above description is merely a preferred embodiment of the present invention and is not intended to limit the invention in any other way. Any person skilled in the art may make changes or modifications to the above-disclosed technical content to create equivalent embodiments. However, any simple modifications, equivalent changes, and modifications made to the above embodiments based on the technical essence of the present invention without departing from the scope of the present invention shall still fall within the protection scope of the present invention.

Claims

1. The soybean GmMADS119 gene, characterized by, The amino acid sequence encoded by the gene is shown in SEQ ID NO:

2.

2. The soybean GmMADS119 gene according to claim 1, characterized in that, The nucleic acid sequence of the gene is shown in SEQ ID NO:

1.

3. The application of the soybean GmMADS119 gene as described in claim 1 in the regulation of plant flowering period.

4. The application according to claim 3, characterized in that, The soybean GmMADS119 gene was constructed into an expression vector, transformed into plants, and its expression level in plants was increased, thereby promoting flowering.

5. The application according to claim 4, characterized in that, The method of plant transformation is one of Agrobacterium-mediated transformation, gene gun transformation, electroporation transformation, PEG transformation, or liposome transformation.

6. The application according to claim 3, characterized in that, The plant in question is either soybean or Arabidopsis thaliana.

7. A method for promoting flowering in Arabidopsis thaliana, characterized in that, The coding region sequence of the soybean GmMADS119 gene was constructed into a plant expression vector, which was then transformed into Arabidopsis thaliana to promote flowering of Arabidopsis thaliana under long-day and short-day conditions; the nucleic acid sequence of the soybean GmMADS119 gene is shown in SEQ ID NO:

1.

8. The method for promoting flowering in Arabidopsis thaliana according to claim 7, characterized in that, The long-day conditions are: 16 hours of light at 23°C and 8 hours of darkness at 20°C; humidity of 55%; light exposure begins at 7:00 AM; and light intensity of 14000 Lx. The short-day conditions are: 10 hours of light at 23°C and 14 hours of darkness at 20°C; humidity of 55%; light exposure begins at 7:00 AM; and light intensity of 14000 Lx.

9. A method for promoting soybean flowering, characterized in that, The coding region sequence of the soybean GmMADS119 gene was constructed into a plant expression vector, transformed into soybean, and expressed in soybean to promote flowering of soybean under long-day and semi-long-day conditions; the nucleic acid sequence of the soybean GmMADS119 gene is shown in SEQ ID NO:

1.

10. The method for promoting soybean flowering according to claim 9, characterized in that, The long-day conditions are: 16 hours of light at 25°C and 8 hours of darkness at 23°C; the start time of light exposure is 8:00 AM; and the light intensity is 14000 Lx. The conditions for the longer daylight hours are: summer outdoor natural temperature, 14.5 hours of sunshine, and 9.5 hours of darkness.

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