A siberian apricot psmads19 gene, a coding protein, a primer set, a recombinant vector and application thereof in regulating flowering time of plants

Abstract

The application discloses Siberian apricot PsMADS19 genes, coding proteins, primer groups, recombination carriers and application thereof in regulating flowering time of plants. A coding sequence of the PsMADS19 gene is shown as SEQ ID NO. 1, and an amino acid sequence of a coding protein of the PsMADS19 gene is shown as SEQ ID NO. 2. The expression amount of the PsMADS19 gene in a transgenic plant is significantly increased, the bolting and flowering time is delayed, and the flowering period can be effectively delayed. The application further discloses specific primers of the PsMADS19 gene and primers for fluorescence quantitative PCR. The application further provides a main gene and a molecular breeding method for late flowering and frost resistance breeding of Siberian apricot and other plants, and has important theoretical value and production application prospect.

Description

Technical Field

[0001] This invention relates to the field of plant genetic engineering technology. Specifically, it relates to a regulatory gene PsMADS19 cloned from Siberian apricot, its encoding protein, primer set, recombinant vector, and the application of this gene in delaying flowering and cultivating late-flowering varieties. Background Technology

[0002] Siberian apricot (Prunus sibirica L.) is a plant belonging to the genus Armeniaca in the subfamily Prunoideae of the family Rosaceae. It is a major tree species cultivated for soil and water conservation forests and economic forests, possessing both ecological and economic value. Ecologically, Siberian apricot is highly adaptable, exhibiting cold resistance, drought resistance, and tolerance to poor soil conditions. It is a pioneer tree species for ecological construction in arid and semi-arid regions of China, capable of greening barren mountains, conserving soil and water, and protecting water resources. Economically, the apricot flesh can be eaten raw and used to make wine; the apricot kernels can be used medicinally and as an industrial raw material; the hard wood can be used for furniture making; the seed shells can be used to make activated carbon; and the leaves can be used as animal feed. However, Siberian apricot yields are severely affected by late frost damage. Therefore, selecting superior individual plants with late-flowering traits to avoid frost damage is key to solving the bottleneck problem in the industry.

[0003] Flowering time is an important agronomic trait in economic forest trees, directly affecting fruit yield and quality. The MCM1-AGAMOUS-DEFICIENS-SRF box (MADS-box) gene family is a highly conserved class of regulatory factors in plants, with members widely involved in flowering induction. In the model plant Arabidopsis thaliana, several MADS-box genes, such as SUPPRESSOR OF OVEREXPRESSION OF CONSTANS 1 (AtSOC1) and FLOWERING LOCUS C (AtFLC), have been shown to be core factors in the flowering regulatory network.

[0004] The AGAMOUS-like 6 (AGL6) subfamily, due to an ancient duplication event during evolution, formed euAGL6 and AGL6-like gene branches at the base of core eudicots, existing in multiple groups of gymnosperms, angiosperms, monocots, and dicots. Studies have shown that AGL6 and its homologous genes have been cloned and their expression patterns analyzed in different species. For example, species such as jujube (Ziziphus jujuba), Cymbidium goeringii, and Cymbidium faberi possess conserved MADS-box domains. Mu et al. found that the ZjAGL6 gene in jujube is highly expressed during flower development, with the highest expression level during the large bud stage. Hu et al. found that the CgAGL6-3 expression in Cymbidium goeringii varies across floral organs, mainly concentrated in the lip. In functional studies, Hu et al.'s research on the CgAGL6 transgenic tobacco phenotype of Cymbidium goeringii × Cymbidium faberi hybrids showed an earlier flowering time.

[0005] Therefore, verifying gene function through biotechnology to delay flowering and avoid late frost has important application value for stable yield breeding of plants such as apricot, peach, and plum. Summary of the Invention

[0006] Therefore, the technical problem to be solved by this invention is to provide a Siberian apricot PsMADS19 gene, its encoded protein, primer set, and its application in regulating plant flowering time and cultivating late-flowering plants. This gene participates in the regulation of plant flowering time, and when the gene is overexpressed, the flowering time of the plant is delayed.

[0007] To solve the above-mentioned technical problems, the present invention provides the following technical solution:

[0008] A PsMADS19 gene for Siberian apricot, the coding sequence of which is shown in SEQ ID NO.1; the PsMADS19 gene is used to delay the flowering period of Siberian apricot.

[0009] The protein encoded by the PsMADS19 gene of the Siberian apricot mentioned above has an amino acid sequence as shown in SEQ ID NO.2, which has 239 amino acids.

[0010] The primer pairs for amplifying the PsMADS19 gene of the Siberian apricot are shown in SEQ ID NO.3 and SEQ ID NO.4, respectively.

[0011] A recombinant vector containing the Siberian apricot PsMADS19 gene, wherein the recombinant vector contains the aforementioned Siberian apricot PsMADS19 gene.

[0012] The above-mentioned application of the Siberian apricot PsMADS19 gene in regulating plant flowering time; the coding sequence of the Siberian apricot PsMADS19 gene was obtained via Gateway. TM The technology is used to construct the entry vector pDONR207, which is then linked to the expression vector pMDC32 to obtain the final expression vector pMDC32-PsMADS19 of the recombinant vector.

[0013] The above-mentioned application of the Siberian apricot PsMADS19 gene in regulating the flowering time of plants refers to using the Siberian apricot PsMADS19 gene to regulate the flowering time of plants.

[0014] The above-mentioned application of the PsMADS19 gene of Siberian apricot in regulating the flowering time of plants: overexpression of the PsMADS19 gene of Siberian apricot is used to delay the flowering time of Siberian apricot.

[0015] The application of the Siberian apricot PsMADS19 gene in regulating plant flowering time was investigated by transforming Arabidopsis thaliana using the inflorescence infection method. RNA was extracted from leaves of wild-type and transgenic Arabidopsis thaliana, and cDNA was obtained by reverse transcription. Using wild-type Arabidopsis thaliana as a control, the expression level of PsMADS19 in transgenic Arabidopsis thaliana was detected by real-time quantitative PCR.

[0016] The above-mentioned application of the Siberian apricot PsMADS19 gene in regulating plant flowering time: Overexpression of the PsMADS19 gene in Arabidopsis thaliana is used to delay bolting and flowering time in Arabidopsis thaliana. To demonstrate this application, real-time quantitative PCR was performed using primers with sequences shown in SEQ ID NO. 5 and SEQ ID NO. 6 to amplify the Siberian apricot PsMADS19 gene in transgenic Arabidopsis thaliana.

[0017] The technical solution of the present invention achieves the following beneficial technical effects:

[0018] 1. The PsMADS19 gene cloned from Siberian apricot provided by this invention is clearly identified as belonging to the MADS-box family and having typical conserved domains;

[0019] 2. The present invention provides that when the PsMADS19 gene from Siberian apricot is overexpressed in plants, it is confirmed that PsMADS19 is a flowering inhibitor and that overexpression of the PsMADS19 gene from Siberian apricot can delay flowering in plants.

[0020] 3. The application of the PsMADS19 gene provides a reproducible system of gene cloning, vector construction, Arabidopsis transformation and phenotypic identification technology, which can be directly used for the breeding of late-flowering varieties.

[0021] 4. Analysis of the expression pattern of the PsMADS19 gene in Siberian apricot showed that its expression level was higher in late-flowering Siberian apricots during the dormant to full-blooming period (S4-S6) than in early-flowering ones. Expression varied across different tissues and floral organs, with the highest expression in flowers, flower buds, and calyxes; and the lowest expression in leaves, pulp, kernels, and stamens. Overexpression of the PsMADS19 gene in Siberian apricots delays the flowering time.

[0022] 5. Overexpression of the PsMADS19 gene in Arabidopsis thaliana delayed bolting by an average of 3 days and flowering by an average of 2 days. Overexpression of the PsMADS19 gene in Arabidopsis thaliana significantly delayed bolting and flowering, and can be used to cultivate late-flowering plants. It negatively regulates the expression of downstream flowering genes AtFT and AtFUL, and positively regulates the expression of AtFLC, thereby controlling the plant's flowering period.

[0023] 6. The PsMADS19 gene is involved in the regulation of flowering time and affects the expression of downstream flowering genes. The results of this study provide a theoretical basis for further exploration of the regulation of flowering time in Siberian apricots. Attached Figure Description

[0024] Figure 1 Electrophoresis diagram of the product obtained by amplifying Siberian apricot cDNA using primers PsMADS19-F and PsMADS19-R in this embodiment of the invention.

[0025] Figure 2 The results of sequence comparison of conserved protein regions of the PsMADS19 gene of Siberian apricot with those of closely related species such as peach, Xueluozhai cherry blossom, forest strawberry, rose, plum, and the model plant Arabidopsis thaliana in this invention.

[0026] Figure 3 The expression levels of PsMADS19 at different time points (A) and in different tissues and floral organs (B); A represents the expression levels of PsMADS19 at different time points, and the test method is two-way ANOVA. Different asterisks indicate * p ≤ 0.05; ** p ≤ 0.01; *** ≤ 0.001; **** p ≤ 0.0001, and NS indicates no significant difference; B represents the expression levels of PsMADS19 in different tissues and floral organs, and the test method is Duncan's test. Different lowercase letters indicate significant differences at the P < 0.05 level.

[0027] Figure 4 35S::PsMADS19 transgenic Arabidopsis thaliana phenotype;

[0028] Figure 5 Identification of 35S::PsMADS19 in Arabidopsis thaliana was performed using an independent samples t-test. Different asterisks indicate p ≤ 0.05; p ≤ 0.01; p ≤ 0.001; p ≤ 0.0001.

[0029] Figure 6 The bolting time (A) / day and flowering time (B) of 35S::PsMADS19 in Arabidopsis thaliana were compared using an independent samples t-test. Different asterisks indicate p ≤ 0.05; p ≤ 0.01; p ≤ 0.001; p ≤ 0.0001.

[0030] Figure 7 Expression patterns of flowering genes in PsMADS19 overexpression transgenic lines; A represents the expression pattern of AtFT in 35S::PsMADS19 transgenic Arabidopsis thaliana; B represents the expression pattern of AtFUL in 35S::PsMADS19 transgenic Arabidopsis thaliana; C represents the expression pattern of AtFLC in 35S::PsMADS19 transgenic Arabidopsis thaliana; The test method was independent samples t-test, with different asterisks indicating * p≤0.05; ** p≤0.01; ***≤0.001; **** p≤0.0001; ns indicates no significant difference. Detailed Implementation

[0031] 1. Experimental materials

[0032] The gene cloning material was Siberian apricot flower buds. The materials used for analyzing the expression pattern of the PsMADS19 gene at different stages were early-flowering (NF) and late-flowering (LF) Siberian apricots, with the late-flowering variety flowering an average of 7.5 days later than the early-flowering variety. The variety was from the Arukorqin 41 family, and was mainly divided into six different developmental stages: early differentiation (S1), mid-differentiation (S2), late differentiation (S3), dormancy (S4), initial flowering (S5), and full bloom (S6). The materials used for analyzing the gene expression patterns of different tissues and floral organs were Siberian apricot flower buds, flowers, leaves, pulp, kernels, stamens, pistils, calyxes, and petals. The heterologous overexpression experimental material was Columbia wild-type Arabidopsis thaliana (Columbia-0, Col-0), cultured under 16 h light / 8 h dark conditions at 22℃ and 70% relative humidity. All samples were immediately placed in liquid nitrogen after collection and stored at -80℃ for subsequent experiments.

[0033] 2. Experimental Methods

[0034] 2.1 Extraction of total RNA from plants and cloning of the PsMADS19 gene

[0035] This study used TRIzol reagent to extract total RNA from Siberian apricot flower buds and Arabidopsis leaves, and analyzed it using HiScript. ® First-strand cDNA was synthesized using the II Q RT SuperMix for qPCR kit. Primers were designed based on the gene CDS sequence using Primer 5.0 software, and the full-length CDS sequence of the PsMADS19 gene was amplified by PCR using Phusion high-fidelity DNA polymerase. The reaction program was: 98℃ pre-denaturation for 30 s, 98℃ denaturation for 10 s, 55℃ annealing for 30 s, and 72℃ extension for 30 s, for a total of 35 cycles; the final extension was 5 min at 72℃. PCR products were detected by 1% agarose gel electrophoresis. The target band was excised and purified using the TIANgel Midi Purification Kit. The purified DNA product was sent to Henan Shangya Biotechnology Co., Ltd. for sequencing to obtain the PsMADS19 gene sequence.

[0036] Table 1 PsMADS19 PCR reaction system

[0037]

[0038] 2.2 Analysis of multiple sequence alignment and conserved domains of the PsMADS19 gene

[0039] Using the gene sequences obtained from the cloned products through first-generation sequencing, the homologous genes of PsMADS19 in peach (Prunus persica), snow cherry (Prunus xueluoensis), forest strawberry (Fragaria vesca), rose (Rosarugosa), Arabidopsis thaliana, and plum (Prunus mume) were compared using DNAMAN software on the NCBI website BLAST. The amino acid sequences of these genes were then imported into the BLAST online analysis software (https: / / www.ncbi.nlm.nih.gov / cdd / ) for predictive analysis of their conserved domains.

[0040] 2.3 Analysis of PsMADS19 gene expression patterns

[0041] Based on RNA-Seq data from early and late flowers of Siberian apricot at six different developmental stages, as well as from different tissues and floral organs, the expression level of PsMADS19 in different time periods, tissues, and floral organs was analyzed.

[0042] 2.4 PsMADS19 gene functional analysis

[0043] 1) Construction of PsMADS19 gene vector

[0044] Use Gateway TM The technology constructs an introductory vector and plant expression vector for the PsMADS19 gene. Gateway vectors are added to both ends of the primers according to the construction requirements. TM The adapter sequence was obtained, and the full-length CDS sequence of the PsMADS19 gene was amplified by PCR technology. The primer design is shown in Table 1.

[0045] The entry vector plasmid (pDONR207) was placed according to Gateway... ® BP Clonase ™ II. Following the instructions of the Enzyme Mix reagent, a BP recombination reaction was performed, transforming the cells into *Escherichia coli* DH5α competent cells. Then, the entry-level recombinant vector (pDONR207-PsMADS19) and expression vector (pMDC32) were introduced according to the Gateway protocol. ® LR Clonase ™ II. Following the instructions of the EnzymeMix reagent, the LR recombination reaction was performed and sequenced to verify the results, yielding the final expression vector pMDC32-PsMADS19.

[0046] 2) Statistical analysis of heterologous transformation of Arabidopsis thaliana and flowering phenotype.

[0047] Arabidopsis thaliana was transformed via inflorescence infection. This was achieved by converting OD... 600 The culture temperature was increased to 0.8, and the bacterial cells were collected after centrifugation. The cells were resuspended in a 5% sucrose solution, and then 0.02% Silwet L-77 surfactant was added to obtain the infection solution. Open inflorescences and siliques were removed before infection, and the infection time was 45 s. After infection, the cells were incubated in the dark at 22°C for 24 h. After disinfection with a 2% sodium hypochlorite solution, the T1 generation seeds were harvested using Hygromycin B (30 mg / L), and the progeny seeds were cultured to the T3 generation under long-day conditions. Under the same growing environment, the bolting time was recorded when the wild-type and transgenic Arabidopsis reached a bolting height of 0.5 cm, and the flowering time was recorded when the first flower opened.

[0048] 3) Positive identification of transgenic Arabidopsis lines and expression analysis of flowering-related genes.

[0049] RNA was extracted from leaves of wild-type and transgenic Arabidopsis thaliana, and cDNA was obtained by reverse transcription. Using wild-type Arabidopsis thaliana as a control, the expression level of PsMADS19 in transgenic Arabidopsis thaliana was detected by real-time quantitative PCR (RT-qPCR). Simultaneously, the expression levels of the Arabidopsis thaliana flowering genes FLOWERING LOCUS T (AtFT), FRUITFULL (AtFUL), and FLC in 35S::PsMADS19 transgenic Arabidopsis thaliana were detected. The reaction program was 95℃ for 30 s; 95℃ for 10 s, 60℃ for 30 s, for 40 cycles; the melting curve was 60℃-95℃ with a temperature increase of 5℃ / s. -ΔΔCt The relative expression levels of genes were calculated using a method with three biological replicates for each sample. Primer designs are shown in Table 2.

[0050] Table 2. Primer sequences for the PsMADS19 gene

[0051]

[0052] Note: The underlined part is Gateway. TM Connector sequence

[0053] 3 Results and Analysis

[0054] 3.1 PsMADS19 gene cloning and protein sequence alignment

[0055] Using the cDNA of the Siberian apricot PsMADS19 gene as a template, PCR amplification yielded the PsMADS19 gene sequence, which is 720 bp in length and encodes 239 amino acids. Figure 1 ).

[0056] PsMADS19 gene sequence (SEQ ID NO. 1):

[0057] ATGGGGAGACGGAAAGTGGTGTTGGGGAGAATAGAAAACAAGATCAACCGTCAAGTGACCTTCTCAAAACGAAGAAATGGTCTGCTCAAGAAAGCCTACGAGCTCTCTGTGCTCTGCGATGCCCAAGTTGCCCTCATCATCTCCTCCAGCCGTGGCAAGCTCTATGAGTTTGGCAGCACTGACGTGAACAAAATTCTTGAGCGGTACCGTCAATGTTGCTACTCCTTACAAGGCAATGTTGCTGAGAATGAAACACAGAACTTATACCAAGAGGTCTCAAAATTAAAGGTCAAATACGAGTCTCTTCAACTTTCACAGAGGCATTTGCTTGGAGAAGATCTTGAAAAGCTTCGCCTGAAAGAACTGGTGAATCTTGAGAATCAACTTGGCAAAACTCTCTCAAAAGCTAGACAACGAAAGACAGAGATGATGTATGATCGATTAGAAGAACTGCGCCAAAAGGAGAACGATCTTGGAGAGAAAAATAAGCAGCTCAAATCCGAGCTTGAGGAAGAAGAGCATGCTCAAGCAACACTGCAAGGTCCGGGAGAGCTTCCACTTACAAAAGACAATCGCATTGAATCAGAGCATCCCAGCTTACGCTTACAGATATGGGATCACCAACCAGCCCCCCAAGAAAAAGCAGCTGATGATGATCAAGGGAGGGCCATGGTGGGTGGGAGCAAGAGTAACTGCAACCAGCGGGGTTGGCTTCTTTAA

[0058] Amino acid sequence of the protein encoded by the PsMADS19 gene (SEQ ID NO. 2):

[0059] MGRRKVVLGRIENKINRQVTFSKRRNGLLKKAYELSVLCDAQVALIISSSRGKLYEFGSTDVNKILERYRQCCYSLQGNVAENETQNLYQEVSKLKVKYESLQLSQRHLLGEDLEKLRLK ELVNLENQLGKTLSKARQRKTEMMYDRLEELRQKENDLGEKNKQLKSELEEEEHAQATLQGPGELPLTKDNRIESEHPSLRLQIWDHQPAPQEKAADDDQGRAMVGGSKSNCNQRGWLL*

[0060] The PsMADS19 gene sequence was compared with homologous gene sequences of model plants and closely related species using DNAMAN software. Sequence analysis showed that all of these genes contain highly conserved MADS and K-box domains. Specifically, the PsMADS19 gene showed similarities of 95.40%, 83.75%, 60.17%, 56.25%, 48.41%, and 44.21% with PpMADS6, PxuMADS1, FveMADS6, RrMADS3, AtAGL6, and PmCMB1, respectively. Therefore, the PsMADS19 gene belongs to a typical member of the MADS-box gene family. Figure 2 ).

[0061] 3.2 Analysis of PsMADS19 gene expression patterns

[0062] Based on transcriptome data from early and late flowering of Siberian apricot at six different developmental stages, as well as from different tissues and floral organs, the expression pattern of the PsMADS19 gene was analyzed. The results showed that the expression level of the PsMADS19 gene was significantly higher in late-flowering than in early-flowering during the dormancy to full bloom period (S4-S6). Its expression level in flower buds and flowers was significantly higher than in leaves, pulp, and kernels. Among floral organs, the expression level was highest in the calyx and lowest in the stamens, suggesting that this gene may be involved in regulating flowering time. Figure 3 ).

[0063] 3.3 PsMADS19 gene functional analysis

[0064] To determine the function of the PsMADS19 gene, the overexpression vector pMDC32-PsMADS19 was constructed. Wild-type Arabidopsis thaliana was transformed using the inflorescence infection method to obtain transgenic plants, and three transgenic lines were screened to obtain transgenic lines. Figure 4 RNA was extracted and used for RT-qPCR validation of the 35S::PsMADS19 transgenic line. Expression analysis showed that the gene expression level in positive plants was significantly upregulated compared to the wild type. Figure 5 ).

[0065] This study further statistically analyzed the bolting time of T3 generation Arabidopsis thaliana lines 35S::PsMADS19 (n=10) and wild-type Arabidopsis thaliana (n=10) cultured under the same conditions. The results showed that the bolting time of wild-type Arabidopsis thaliana under long-day conditions was 21.7±1.3 days. The bolting times of transgenic Arabidopsis thaliana lines 35S::PsMADS19#1, 35S::PsMADS19#2, and 35S::PsMADS19#3 were 24.5±1.7 days, 24.8±1.0 days, and 24.7±1.6 days, respectively. The bolting time of the 35S::PsMADS19 transgenic Arabidopsis thaliana was significantly delayed compared to wild-type Arabidopsis thaliana. Figure 6 A).

[0066] The flowering time of wild-type Arabidopsis thaliana was 25.5±1.1 days, while the flowering times of 35S::PsMADS19 transgenic Arabidopsis thaliana were 27.5±1.4 days, 27.8±0.8 days, and 27.4±1.2 days, respectively. Figure 6 B), whose flowering time is significantly delayed compared to wild-type Arabidopsis thaliana.

[0067] The above results indicate that overexpression of the PsMADS19 gene in *Arabidopsis thaliana* can delay flowering. Figure 4 ).

[0068] In plants, FT, FUL, and FLC are important for regulating flowering time. Therefore, this study selected AtFT, AtFUL, and AtFLC and analyzed their expression patterns in positive 35S::PsMADS19 transgenic Arabidopsis plants. Figure 7 (As shown). The results showed that the relative expression levels of flowering-promoting genes AtFT and AtFUL in 35S::PsMADS19 transgenic Arabidopsis plants were downregulated compared to the wild type, while the relative expression level of flowering-inhibiting gene AtFLC was upregulated compared to the wild type. Specifically, the expression levels of AtFT in 35S::PsMADS19 transgenic Arabidopsis plants were 0.28, 0.28, and 0.73 times that of the wild type, respectively; AtFUL was 0.57, 0.86, and 0.40 times that of the wild type; and AtFLC was 21.12, 6.11, and 8.55 times that of the wild type. Figure 7 ).

[0069] Obviously, the above embodiments are merely illustrative examples for clear explanation and are not intended to limit the implementation. Those skilled in the art will recognize that other variations or modifications can be made based on the above description. It is neither necessary nor possible to exhaustively list all possible implementations here. However, obvious variations or modifications derived therefrom are still within the scope of protection of the claims of this patent application.

Claims

1. A Siberian apricot PsMADS19 gene, characterized in that, The coding sequence of the PsMADS19 gene of Siberian apricot is shown in SEQ ID NO.1; the PsMADS19 gene is used to delay the flowering period of Siberian apricot.

2. The protein encoded by the Siberian apricot PsMADS19 gene as described in claim 1, characterized in that, The amino acid sequence of the protein encoded by the PsMADS19 gene is shown in SEQ ID NO.2, and has 239 amino acids.

3. The primer pair for amplifying the Siberian apricot PsMADS19 gene as described in claim 1, characterized in that, The nucleotide sequences of the forward and reverse primers for amplifying the PsMADS19 gene are shown in SEQ ID NO.3 and SEQ ID NO.4, respectively.

4. A recombinant vector containing the Siberian apricot PsMADS19 gene, characterized in that, The recombinant vector contains the Siberian apricot PsMADS19 gene as described in claim 1.

5. The recombinant vector of the Siberian apricot PsMADS19 gene according to claim 4, characterized in that, The coding sequence of the Siberian apricot PsMADS19 gene was obtained via Gateway. TM The technology is used to construct the entry vector pDONR207, which is then linked to the expression vector pMDC32 to obtain the final expression vector pMDC32-PsMADS19 of the recombinant vector.

6. The application of the Siberian apricot PsMADS19 gene as described in claim 1 in regulating plant flowering time, characterized in that, The application involves using the PsMADS19 gene of Siberian apricot to regulate the flowering time of the plant.

7. The application of the Siberian apricot PsMADS19 gene in regulating plant flowering time according to claim 6, characterized in that, Overexpression of the PsMADS19 gene in Siberian apricot was used to delay the flowering time of Siberian apricot.

8. The application of the Siberian apricot PsMADS19 gene according to claim 6 in regulating plant flowering time, characterized in that, Arabidopsis thaliana was transformed using the inflorescence infection method. RNA was extracted from leaves of wild-type and transgenic Arabidopsis thaliana, and cDNA was obtained by reverse transcription. Using wild-type Arabidopsis thaliana as a control, the expression level of PsMADS19 in transgenic Arabidopsis thaliana was detected by real-time quantitative PCR.

9. The application of the Siberian apricot PsMADS19 gene according to claim 8 in regulating plant flowering time, characterized in that, Overexpression of the PsMADS19 gene in Arabidopsis thaliana can delay bolting and flowering time.

10. The application of the Siberian apricot PsMADS19 gene according to claim 8 in regulating plant flowering time, characterized in that, When performing real-time quantitative PCR, primers with sequences shown in SEQ ID NO. 5 and SEQ ID NO. 6 were used to amplify the Siberian apricot PsMADS19 gene in transgenic Arabidopsis thaliana.

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