PoCRK27 gene and related biological material in regulating petal senescence of plants

Abstract

The application belongs to the technical field of plant genetic engineering, and particularly relates to application of PoCRK27 gene and related biological materials thereof in regulation of petal senescence of plants. The PoCRK27 gene encodes a protein with an amino acid sequence as shown in SEQ ID NO:1. The application firstly determines the positive role of the PoCRK27 gene in regulation of petal senescence of peony, which not only deepens the understanding of functions of the CRK gene family of plants, but also provides an important theoretical basis and gene resources for improving storage resistance and ornamental period of ornamental plants such as peony through molecular design.

Description

Technical Field

[0001] This invention belongs to the field of plant genetic engineering technology, specifically involving the application of the PoCRK27 gene and related biomaterials in regulating the senescence of plant petals. Background Technology

[0002] Peonies possess extremely high ornamental, cultural, and medicinal value, and the opening and senescence of their petals directly impact their economic value and ecological benefits. Therefore, studying the relevant molecular mechanisms of peony petal senescence is crucial for improving industrial economic efficiency and promoting the development of green agriculture.

[0003] Cysteine-rich receptor-like kinases (CRKs) are a large subfamily of protein kinases in plants. Their most notable feature is the presence of one or more conserved DUF26 (Domain of Unknown Function 26, PF01657) domains in their extracellular regions. Each DUF26 motif typically contains a highly conserved “C-X8-C-X2-C” cysteine ​​motif. This structure may activate downstream signaling pathways by forming zinc finger structures or disulfide bonds to bind specific metal ions, or by participating in protein interactions.

[0004] CRK plays a crucial role in regulating various life activities such as plant growth and development, signal transduction, and stress response, but systematic research on this family of genes in peony, an important economic crop, is still lacking. Summary of the Invention

[0005] To address the shortcomings of the existing technologies, the purpose of this invention is to provide the application of the PoCRK27 gene and related biomaterials in regulating plant petal senescence.

[0006] To achieve the above objectives, the present invention adopts the following technical solution: In a first aspect, the present invention provides the application of the PoCRK27 gene and related biological materials in regulating the senescence of plant petals, wherein the PoCRK27 gene is a protein encoding an amino acid sequence as shown in SEQ ID NO:1; and the plant is Arabidopsis thaliana or Paeonia suffruticosa.

[0007] Furthermore, the nucleotide sequence of the PoCRK27 gene is shown in SEQ ID NO:2.

[0008] Furthermore, the regulation of plant petal senescence involves: using genetic engineering techniques to overexpress the PoCRK27 gene in plants to delay the senescence time of plant petals and prolong the flowering period.

[0009] Furthermore, the biological material includes recombinant DNA, expression cassettes, transposons, plasmid vectors, viral vectors, engineered bacteria, transgenic cell lines, or non-renewable plant parts.

[0010] A second aspect of the present invention provides a method for delaying the senescence of plant petals, the method comprising: overexpressing the PoCRK27 gene in a plant using genetic engineering techniques; wherein the PoCRK27 gene is a protein encoding an amino acid sequence as shown in SEQ ID NO:1.

[0011] Furthermore, the plant in question is Arabidopsis thaliana or Paeonia suffruticosa.

[0012] Furthermore, the overexpression mode is selected from the following (1) to (4), or any combination thereof: (1) By introducing a plasmid containing the PoCRK27 gene; (2) By increasing the copy number of the PoCRK27 gene on plant chromosomes; (3) By operatively linking a strong promoter to the PoCRK27 gene; (4) By introducing enhancers.

[0013] Expression vectors carrying the target gene can be introduced into plant cells using conventional biotechnological methods such as Ti plasmids, plant virus vectors, direct DNA transformation, microinjection, and electroporation (Weissbach, 1998, Method for Plant Molecular Biology VIII, Academy Press, New York, p. 411). 463 pages; Geiserson and Corey, 1998, Plant Molecular Biology, 2nd Edition).

[0014] A third aspect of the present invention provides the application of transgenic plants obtained by the method described in the second aspect in plant breeding.

[0015] Furthermore, the purpose of the breeding is to delay the senescence of plant petals and prolong the flowering period.

[0016] Furthermore, breeding methods include transgenic, hybridization, backcrossing, self-pollination, or asexual reproduction.

[0017] Compared with the prior art, the technical solution of the present invention has the following beneficial effects: This invention provides the first systematic identification and analysis of the CRK gene family at the whole-genome level in peony, clarifying the positive role of PoCRK27 in regulating peony petal senescence. Silencing PoCRK27 significantly accelerates petal senescence, indicating that the PoCRK27 gene is a key positive regulator of peony petal senescence and abscission. This discovery not only deepens our understanding of the function of the plant CRK gene family but also provides important theoretical basis and genetic resources for improving the storage tolerance and ornamental period of ornamental plants such as peonies through molecular design. Attached Figure Description

[0018] Figure 1 This diagram illustrates the specific expression pattern of the PoCRK family in petal tissues during the flowering process.

[0019] Figure 2 The results show the expression of the PoCRK04 gene in different tissues of peony.

[0020] Figure 3 The results show the expression of the PoCRK08 gene in different tissues of peony.

[0021] Figure 4 The results show the expression of the PoCRK21 gene in different tissues of peony.

[0022] Figure 5 The results show the expression of the PoCRK27 gene in different tissues of peony.

[0023] Figure 6 Subcellular localization results of the PoCRK27 gene.

[0024] Figure 7 The results are for the BIFC interaction validation of the PoCRK27 gene.

[0025] Figure 8 The results show the promoter expression analysis of PoCRK27, where A and B are the expression analyses in Arabidopsis inflorescences and pods, respectively.

[0026] Figure 9 The results show the abscission phenotype and relative expression level of PoCRK27 in the 35S:PoCRK27 transgenic Arabidopsis thaliana. In this figure, A represents the abscission phenotype of the 35S:PoCRK27 transgenic Arabidopsis thaliana, and B represents the relative expression level of PoCRK27.

[0027] Figure 10 The figures show the relative expression level of the AtBOP gene and the petal tearing force of the 35S:PoCRK27 transgenic plants. In the figure, A represents the relative expression level of the AtBOP gene, and B represents the petal tearing force. In the figure, * indicates a significant difference (P<0.05), and ** indicates an extremely significant difference (P<0.01).

[0028] Figure 11 Phenotypic diagram of petal drop in pTRV-PoCRK27 plants.

[0029] Figure 12 This is a comparison of petal shedding time between the virus-induced silent PoCRK27 experimental group and the control group. Figure 13 The results show the malondialdehyde (MDA) and proline content in the virus-induced silencing PoCRK27 experimental group and the control group. In the figure, A represents the MDA content and B represents the proline content.

[0030] Figure 14 Paraffin sections of the detachment area and both sides of the virus-induced silent PoCRK27 experimental group and control group at 24 h, 36 h, 48 h, 60 h and 72 h are shown in the figures. In the figures, R represents the receptacle, P represents the petal, and AZ represents the detachment area. Detailed Implementation

[0031] It should be noted that the following detailed descriptions are exemplary and intended to provide further illustration of the invention. Unless otherwise specified, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention pertains.

[0032] It should be noted that the terminology used herein is for the purpose of describing particular embodiments only and is not intended to limit the exemplary embodiments of the present invention. As used herein, the singular form is intended to include the plural form as well, unless the context clearly indicates otherwise. Furthermore, it should be understood that when the terms "comprising" and / or "including" are used in this specification, they indicate the presence of features, steps, operations, and / or combinations thereof.

[0033] To enable those skilled in the art to better understand the technical solution of the present invention, the technical solution of the present invention will be described in detail below with reference to specific embodiments.

[0034] Description of the main experimental materials and reagents used in the examples: Experimental materials: Peony plant 'Snow Reflecting Peach Blossoms' ( Paeonia suffruticosa 'Xueyingtaohua'), wild-type Colombian Arabidopsis thaliana ( Arabidopsis thaliana Columbia-0) and Bunsen ( Nicotiana benthamiana ).

[0035] Experimental reagents: Restriction endonucleases SmaI, HindIII, SpeI, AscI, NdeI, XhoI, SacI, BamHI, PstI, EcoRI, XbaI. Main reagents: LB Broth Powder, LB Agar Powder (Shanghai Sangon Biotech), Plant RNA Extraction Kit, ChamQ Universal SYBR qPCR Master Mix, Prime STAR® Max DNA Polymerase (Takara), FastPure® Plasmid Mini Kit, FastPure Gel DNA Extraction Mini Kit, HiScript® III RT SuperMix for qPCR, ClonExpress Ultra One Step Cloning Kit, 2×Rapid Taq Master Mix (Novizan). DIG RNA Labeling Kit (SP6 / T7) (Roche, USA), DIG Nucleic Acid Detection Kit (Roche, USA), NBT / BCIP Staining Solution (Roche, Germany), Kanamycin Sulfate, Rifampicin Storage Solution, Acetosyringone (AS), Minimal SD Agar Base, Nutrient-Deficient Culture Media (DOSupplement –Leu / –Trp, DO Supplement –His / –Leu / –Trp, DO Supplement –Ade / –His / –Leu / –Trp, Takara), MgCl2, Ampillin Sodium Salt Storage Solution, Dimethyl Sulfoxide (DMSO) (Beijing Coollife), 2-(N-Morpholinoethanesulfonic Acid) (MES), Silwet L-77 Surfactant (Solarbio).

[0036] The subcellular localization vector pCAMBIA1300-GFP, promoter heterologous expression vector pCAMBIA1300-35S-GUS, Arabidopsis thaliana overexpression transformation vector pCAMBIA1300-35S, bimolecular fluorescence complementary (BIFC) vectors pSPYCE(M) and pSPYNE173, and virus-induced silencing (VIGS) vectors pTRV1 and pTRV2 were obtained from the Key Laboratory of Native Precious Tree Species Breeding of the Shandong Academy of Forestry Sciences, State Forestry and Grassland Administration. They are available to the public from the Institute of Crop Research of the Shandong Academy of Forestry Sciences. These biological materials are only for repeating the relevant experiments of this invention and should not be used for other purposes.

[0037] Escherichia coli (DH5α, catalog number: DLC114) competent cells and Agrobacterium tumefaciens competent cells (GV3101, catalog number: DLC301, EHA105, catalog number: DLC303) were purchased from Tsingke Biotechnology (Beijing).

[0038] Example 1 Gene Expression Analysis The peony flowering period was divided into five stages: bud stage (S1), initial opening stage (S2), full bloom stage (S3), withering stage (S4), and abscission stage (S5). The outermost 3-4 layers of petals and receptacle of the peony flower were cut off together, and a 2 mm × 2 mm tissue sample was taken from the connecting area as the isolation zone. Two 2 mm × 2 mm areas were then taken from this area towards the petals and receptacle as sample regions. Transcriptome data were measured at the five stages to analyze the expression patterns of the peony PoCRK family during the flowering process. A gene expression matrix was constructed based on FPKM (Fragments Per Kilobase of transcript per Million mapped reads) values. The expression data were visualized using the pheatmap package in R. To better compare the expression trends of different genes at each stage, the data were Z-score normalized by gene (row) before heatmap creation.

[0039] The results are as follows Figure 1As shown, PoCRK family members are specifically expressed during the flowering process of peony. PoCRK21 and PoCRK24 are significantly highly expressed in stage S1; PoCRK05 is relatively high in stage S2; and PoCRK27 and PoCRK08 are significantly highly expressed in stage S3. However, during the wilting stage, the expression of all PoCRK family members is low. During the abscission stage (S5), PoCRK04 and PoCRK17 are highly expressed. This specific expression pattern of the PoCRK family during the flowering process suggests that members have different functions at different stages. Highly expressed members during the initial blooming and full bloom stages may be involved in the regulatory pathway of petal opening, while highly expressed members during abscission may be related to senescence and the formation of the protective layer after fracture.

[0040] To further screen candidate genes playing a role in petal senescence, four members—PoCRK04, PoCRK21, PoCRK08, and PoCRK27—that were highly expressed during the flowering period were selected for tissue-specific expression studies. Ten tissues were collected from the 'Snowy Peach Blossom' peony: newly sprouted buds, roots, stems, leaves, petals, receptacle, bracts, sepals, pistils, and stamens during full bloom. Total RNA was extracted and reverse transcribed into cDNA. Quantitative primers were designed using NCBI Primer-BLAST, with beta-actin and β-actin as internal reference genes (PoActin). Detection was performed using SYBR Green I chimeric fluorescence assay on a real-time quantitative PCR instrument. Each sample was configured with three biological replicates and three technical replicates. Relative expression levels were measured using 2-1... -ΔΔCt The primer sequences are shown in Table 1.

[0041] Table 1 Primer Sequences

[0042] like Figures 2 to 5 As shown, the four genes (PoCRK04, PoCRK08, PoCRK21, and PoCRK27) exhibited significant tissue-specific expression. Figure 2 and Figure 3 As shown, PoCRK04 and PoCRK08 exhibit similar expression patterns, primarily concentrated in bracts, sepals, and leaves; as Figure 4 As shown, PoCRK21 is expressed at high levels in bracts, sepals, and pistils; Figure 5 As shown, PoCRK27 is highly expressed in petals, leaves, and receptacles, and this differential expression provides evidence that they perform specific functions in different organs. Therefore, the PoCRK27 gene was selected for subsequent experiments.

[0043] Example 2: Subcellular localization of the PoCRK27 gene The PoCRK27 gene (SEQ ID NO:2) was cloned into the pCAMBIA1300-GFP vector via homologous recombination using the target protein fusion GFP method to construct an expression vector with C-terminal GFP fusion. Subcellular localization of the PoCRK27:GFP fusion protein was then observed. The nucleotide sequence of the PoCRK27 gene is shown in SEQ ID NO:2, and the amino acid sequence of its encoded protein is shown in SEQ ID NO:1. The constructed primer sequences are shown in Table 2.

[0044] (SEQ ID NO:1).

[0045]

[0046] Table 2 Primers for constructing subcellular localization vectors

[0047] For details on the specific steps of Agrobacterium transformation and tobacco infection, please refer to the following references: Li Lan. Genetic and biochemical analysis of the interaction between Arabidopsis thaliana receptor protein kinase TMK and FER and its mediating ABA signaling [D], 2020. Kong Lingyao. Analysis of the regulatory mechanism of Arabidopsis thaliana protein phosphatase ABI1 [D]. 2014.

[0048] Nicotiana Bunsenata leaves were transiently transformed using Agrobacterium-mediated transformation, co-transformed with the nuclear marker protein GHD7-CFP and the membrane marker protein PIP2A-mCherry. After 72 hours of post-infection culture, samples were collected and fluorescence signals were detected using a Leica TCSSP8 Scan Head (Germany) laser copolymer fluorescence microscope. GFP excitation light was 488 nm, and detection light was 510 nm; CFP excitation light was 405 nm, and detection light was 450 nm; mCherry excitation light was 587 nm, and detection light was 610 nm.

[0049] The results are as follows Figure 6 As shown, the fluorescence signal of the PoCRK27:GFP fusion protein co-localizes with the signal of the cytoplasmic membrane marker PIP2A-mCherry, confirming that PoCRK27 is a membrane-localized protein.

[0050] Example 3: Detection of PoCRK27 protein dimerization (BiFC) The ORF sequence of the PoCRK27 gene was cloned into the pSPYCE(M) and pSPYNE173 vectors, respectively. The primer sequences are shown in Table 3. Vector pairs for homodimer detection were constructed.

[0051] Table 3 Primer sequences for homodimer detection

[0052] Tobacco leaves were co-transformed with Agrobacterium. After infection, the leaves were cultured in the dark for 1 day and then in light for 2 days. Temporary sections were prepared by tearing off tobacco leaves with forceps and injecting the lower epidermis. Water was added to the sections, and the sections were laid flat on a glass slide. PIP2A was used as a cell membrane localization protein. The distribution of fluorescence in the cells was observed under 527 nm excitation light. Fluorescence signals were detected using a laser co-fluorescence microscope (Leica TCS SP8 Scan Head (Germany)). YFP excitation light was 510 nm, and detection light was 530 nm; mCherry excitation light was 587 nm, and detection light was 610 nm.

[0053] CRK proteins often function in a dimer form. BiFC experimental results are as follows... Figure 7 As shown, PoCRK27 can form homodimers. The fluorescence signal localization pattern of its dimer is consistent with the subcellular localization results: the PoCRK27 dimer is mainly localized on the cell membrane.

[0054] Example 4: PoCRK27 gene promoter activity analysis The promoter sequence of the PoCRK27 gene (2000 bp upstream of ATG) was cloned and constructed into the pCAMBIAI1300-35S-GUS vector via homologous recombination. Primer sequences are shown in Table 4.

[0055] Table 4 Primer sequences for vector construction

[0056] Arabidopsis thaliana was transformed using the flower immersion method, and T3 generation homozygous transgenic plants were screened. Samples were taken for GUS histochemical staining, and after destaining, the tissue-specific distribution of GUS signals was observed under a stereomicroscope, with a focus on the expression of the PoCRK27 promoter in Arabidopsis thaliana flower organs.

[0057] like Figure 8 As shown in A and B, the expression of the GUS protein signal initiated by Pro-PoCRK27 in the floral organs of Arabidopsis thaliana is specific. The expression of PoCRK27 is mainly concentrated in the floral organs. It is almost not expressed in the newly emerging inflorescences. As the flower develops, it is mainly concentrated in the sepals with a higher expression level. As the flower opens, it is significantly expressed in the petals, and there is also significant expression in the filaments of the stamens. After the petals fall off, it is mainly concentrated at the junction of the fallen fruit and the pedicel. The expression in Arabidopsis thaliana indicates that it plays a role in the floral organs.

[0058] Example 5: Observation of 35S:PoCRK27 transformation of Arabidopsis thaliana and abscission phenotype. The PoCRK27 gene sequence (SEQ ID NO:2) was constructed into the pCAMBIA1300-35S vector via homologous recombination. The primer sequences are shown in Table 5.

[0059] Table 5 Primers for Subcellular Localization Vector Construction

[0060] Transgenic Arabidopsis plants were obtained using the flower immersion transformation method. The T3 generation of transgenic Arabidopsis was used as observation material. Fifteen days after transplanting, leaves were collected, ground, and DNA was extracted from wild-type and 35S:PoCRK27 transgenic plants using the Solarbio Plant Genomic DNA Extraction Kit for identification. The first white flower on the Arabidopsis inflorescence was marked as position 1, and this marking continued downwards. RNA was extracted from positions 5 and 6 of the flowers in 15 positive plants to detect the expression level of PoCRK27. The abscission process of Arabidopsis flower organs was photographed and recorded to comprehensively analyze the effect of heterologous overexpression of PoCRK27 on petal abscission. Quantitative analysis showed that transgenic lines 1, 4, 13, and 15 had higher PoCRK27 expression levels. Lines with higher expression levels were selected for observation. Phenotypic analysis of flower organ abscission revealed that... Figure 9 As shown in Figures A and B, the control group almost completely sheds its flower organs at position 8, while the overexpressing plants shed theirs around position 12. Based on these results, it can be preliminarily inferred that the peony PoCRK27 gene can delay the shedding of Arabidopsis flower organs.

[0061] Expression levels of the abscission indicator gene and petal tearing strength in 35S:PoCRK27 transgenic plants: The expression of the abscission indicator gene AtBOP in inflorescence positions 4, 5, and 6 of the transgenic lines and control plants was determined. Primer sequences are shown in Table 6. Petal tearing strength (pBS) was quantified as the pulling force required to pull a petal from a flower, determined using a digital tensile tester (model: HF-2, Lunjie Electromechanical Instrument Co., Ltd., Shanghai). The petal tearing force was measured in flowers at positions 4, 5, and 6 from top to bottom in inflorescences of control and transgenic plants, with a total of 30 petals randomly measured at each position.

[0062] Table 6 Primer sequences for determining the expression of the exfoliation indicator gene AtBOP.

[0063] The results of the AtBOP gene expression level analysis are as follows: Figure 10 As shown in Figure A, the expression level of this gene at position 5 was found to be significantly higher than that of the transgenic gene, suggesting that this is the critical period for the initiation of abscission in the control group. The petal tearing force data analysis results are as follows... Figure 10 As shown in B, the tearing force of the experimental group and the control group at position 4 was not significantly different. However, at positions 5 and 6, the tearing force of the control group was significantly smaller than that of the experimental group, especially at position 6, where the difference was extremely significant. It is speculated that the petals of the control group were about to fall off at this time, while the overexpression of PoCRK27 delayed the fall off of the flower organs.

[0064] Example 6: Determination of petal drop rate and physiological indicators of pTRV-PoCRK27 Using virus-induced silencing technology, the shedding of peony petals was observed. VIGS primers for PoCRK27 were designed using NCBI (Primer designingtool (nih.gov)) and the specific fragment was constructed onto the VIGS vector (pTRV1). The primer sequences are shown in Table 7.

[0065] Table 7 Primer sequences for silencing vector construction

[0066] The plant material used in the experiment was a robust peony plant of the variety 'Xueyingtaohua' (Paeonia suffruticosa 'Xueyingtaohua'), with petals that were normally developed and prone to falling off due to aging under natural conditions.

[0067] Select 30 four-year-old peony plants. From each plant, select 5 cut peony flowers with buds in roughly the same state. When the outer 1-2 whorls of petals are slightly open, cut the flowers from the mother plant, retaining a stem length of 25-30 cm. Place them in deionized water and quickly transfer them to the laboratory. In the laboratory, underwater, make a 2 cm oblique cut at the end of the flower stem to ensure proper water absorption and prevent clogging. Place them in deionized water and then in a 20℃ light incubator (16 h light / 8 h darkness) at 50% relative humidity. Take peony buds and prick a small hole at the very tip with a needle beforehand. Place the treated peony buds upside down in a beaker containing the infiltration solution. Immerse the entire bud in the solution, place the beaker in a vacuum pump to remove air at 0.1 atmospheres for 20 minutes, slowly release the air, and repeat 3 times. Rinse twice with tap water and once with sterile water. The samples were placed in the dark at 8℃ for 3 days, and then the temperature of the climate chamber was adjusted to 20℃ (16 h light / 8 h dark) and the relative humidity was 50%. Photos were taken every 12 hours to observe changes in petal morphology and record the time of petal shedding. The pTRV1 empty vector was used as a control group. Each control group and experimental group had 30 samples.

[0068] like Figure 11 and Figure 12 As shown, virus-induced silencing of PoCRK27 significantly accelerated the petal shedding process. The complete petal shedding time in the control group was 123.45 h, while in the silent experimental group it was 80.75 h, which was 25.15% earlier than the control group.

[0069] Simultaneously, the contents of malondialdehyde and proline in the control and experimental groups were measured at 12 h, 24 h, 48 h, 72 h, and 84 h after flowering. Figure 13As shown in Figures A and B, the malondialdehyde (MDA) content in the control group was higher at 48 h, then decreased rapidly as the petals aged, reaching a peak at 72 h before declining again. Similarly, the proline content in the control group was highest at 72 h, while the transgenic group remained at a lower level within the measurement range, increasing rapidly at 84 h, presumably indicating the onset of senescence at this point, with petal senescence causing a rapid increase in proline content.

[0070] Example 8: Observation of changes in the absorptive zone of pTRV-PoCRK27 plants via paraffin sections VIGS test samples were transferred to a 20℃ climate chamber for 24 h, 36 h, 48 h, 60 h, and 72 h. Following the sampling method in Example 1, samples were taken from the isolated areas of pTRV-PoCRK27 plants and control plants, and paraffin sections were prepared to observe changes in the isolated areas. Results are as follows: Figure 14 As shown, paraffin sections revealed no significant differences in the connection between petals and receptacle in the control group at 24 h and 36 h. At 36 h, the isolated cells in the silent group had already differentiated (cells were tightly packed, smaller, and had larger nuclei). At 48 h, the isolated cells in the control group began to differentiate, while the intercellular spaces in the silent group had significantly increased. At 60 h, the intercellular spaces in the control group had increased somewhat, while the intercellular spaces in the silent group continued to widen. By 72 h, the intercellular spaces in the silent group had essentially formed a continuous fracture layer, indicating that the petals were about to fall off. The paraffin sections of the isolated area clearly demonstrated that TRV-PoCRK27 accelerated the formation and separation of isolated cells, thus speeding up the petal shedding process.

[0071] Finally, it should be noted that the above description is merely a preferred embodiment of the present invention and is not intended to limit the present invention. Although the present invention has been described in detail with reference to the foregoing embodiments, those skilled in the art can still modify the technical solutions described in the foregoing embodiments or make equivalent substitutions for some of them. Any modifications, equivalent substitutions, improvements, etc., made within the spirit and principles of the present invention should be included within the protection scope of the present invention.

Claims

1. The application of the PoCRK27 gene and its biomaterials in regulating plant petal senescence, characterized in that, The PoCRK27 gene encodes a protein with the amino acid sequence shown in SEQ ID NO:1; the plant is Arabidopsis thaliana or Paeonia suffruticosa. The regulation of plant petal senescence involves overexpressing the PoCRK27 gene in plants using genetic engineering techniques to delay the senescence time of plant petals and prolong the flowering period. The biological materials include recombinant DNA, expression cassettes, transposons, plasmid vectors, viral vectors, and engineered bacteria.

2. The application as described in claim 1, characterized in that, The nucleotide sequence of the PoCRK27 gene is shown in SEQ ID NO:

2.

3. A method for delaying the senescence of plant petals, characterized in that, The method includes: overexpressing the PoCRK27 gene in plants using genetic engineering techniques; the PoCRK27 gene is a protein encoding an amino acid sequence as shown in SEQ ID NO:1; The plant in question is either Arabidopsis thaliana or Paeonia suffruticosa.

4. The method as described in claim 3, characterized in that, The overexpression method is selected from the following (1) to (3), or any combination thereof: (1) By introducing a plasmid containing the PoCRK27 gene; (2) By increasing the copy number of the PoCRK27 gene on plant chromosomes; (3) By operatively linking a strong promoter to the PoCRK27 gene.

Images