Peony PoLEC1 gene and application of protein coded by peony PoLEC1 gene

Abstract

The invention relates to the field of gene editing, in particular to a peony PoLEC1 gene and application of protein coded by the peony PoLEC1 gene. The peony PoLEC1 gene is cloned from cultivated peony paeonia ostii, after the peony PoLEC1 gene is transformed into arabidopsis thaliana, phenotypic contrast finds that the peony PoLEC1 gene can play a role in bolting and flowering as well as growth and development of plants, specifically, overexpression of the peony PoLEC1 gene can increase the number of leaves, delay bolting and flowering time and reduce the height of the plants. According to the invention, arabidopsis thaliana is taken as a model plant, and a certain theoretical basis is provided for establishing a complete plant regeneration and genetic transformation system for peony by exploring the functional characteristics of the peony PoLEC1 gene and the expression characteristics of the peony PoLEC1 gene in the whole somatic embryo direct generation process and in each tissue of overexpressed arabidopsis thaliana.

Description

Technical Field

[0001] The present invention relates to the field of gene editing, and in particular to the application of the peony PoLEC1 gene and the protein encoded by it. Background Art

[0002] The LEAFY COTYLEDON (LAFL) regulatory network consists of four genes (LEC1, ABI3, FUS3, and LEC2), which encode transcription factors that are major regulators of seed development, including oil accumulation and embryonic development.

[0003] LEC1 belongs to the NF-YB subfamily of the NF-Y nuclear transcription factor family, which is a conserved heterotrimeric complex that is ubiquitous in eukaryotes. NF-Y transcription factors are composed of three main subunits, NF-YA, NF-YB, and NF-YC, which bind to the CCAAT box in the promoter region of genes and exert their functions through the formation of heterodimers, forming a diverse subfamily. As a member of the NF-YB subfamily, LEC1 itself does not bind directly to DNA, but interacts with NF-YA and NF-YC, which have DNA binding ability, to form a complex that can recognize and bind to the CCAAT cis-element at the transcription start point. NF-YB and NF-YC subunits have a unique histone fold domain (HFD / HFM), which is formed by three α helices and two loops, which is essential for protein-DNA and protein-protein interactions. LEC1 is also a homolog of the HAP3 (Hemeactivated protein 3) subunit, and HAP3 is most conserved in the B region, which is the determining factor for the function of LEC1.

[0004] LEC genes are involved in plant growth and development, somatic embryogenesis, and the accumulation of storage materials. The induction of spontaneous somatic embryogenesis is due to the ectopic expression of transcription factor genes. LEC, as a key regulatory factor for plant cell totipotency, controls the initiation and regulation of the maturation stage during embryonic development, thereby promoting the generation of somatic embryos. LEC1 was first isolated and identified from Arabidopsis thaliana and is an important regulator of embryonic development. It can activate gene transcription required for embryonic morphogenesis and cell differentiation. Ectopic overexpression of AtLEC1 can induce the formation of embryo-like structures on Arabidopsis leaves and seedling development abnormalities. Stone et al. found that LEC1 promotes embryonic development by forming an environment conducive to the coordinated expression of genes related to embryonic morphological development and maturation. DcLEC1 was isolated from carrot (Daucus carota). In situ hybridization analysis showed that it was expressed in embryonic cells and somatic embryos, but not in endosperm, indicating that DcLEC1 plays a key regulatory role in the development of zygotic and somatic embryos. Zhang et al. showed that the expression pattern of ZmLEC1 in maize (Zea mays L.) somatic embryo development was similar to that in Arabidopsis zygotic embryos, confirming its important role in inducing zygotic and somatic embryogenesis. Cassava (Manihot esculenta) overexpressing MeLEC1 had morphological changes such as fleshy cotyledons, swollen hypocotyls, abnormally shaped leaves, and green roots. MeLEC1 was expressed at a high level at the initial and later stages of somatic embryo induction. The transfer of Selaginella moellendorffii SmLEC1 into the Arabidopsis lec1 mutant revealed that it was mainly accumulated in sexual and asexual reproductive structures, indicating that it was involved in cellular processes similar to those during seed development and maturation. In soybean (Glycine max) embryos, GmLEC1 transcriptionally activated genes encoding light response components of photosystems I and II as well as a group of other genes involved in photosynthesis and chloroplast biogenesis. In rice (Oryza sativa L.), OsLEC1 controls endosperm development by interacting with the AP2 transcription factor. However, no reports have been found on LEC1 in peony.

[0005] In order to further understand the regulatory network during peony somatic embryogenesis, this paper, based on the previous research on key genes of peony and other plant somatic embryos, further explores the expression characteristics of peony LEC genes during embryonic development and their role in plant growth and development, and reveals the molecular regulatory network between these key genes. Through these studies, it is intended to provide theoretical support and technical guidance for the construction of an efficient and stable peony regeneration system and genetic transformation system. Summary of the invention

[0006] The purpose of the present invention is to provide an application of the peony PoLEC1 gene and the protein encoded by it to solve the problems existing in the above-mentioned prior art. The present invention uses Arabidopsis thaliana as a model plant, and by exploring the expression characteristics of the peony PoLEC1 gene in various tissues of overexpressed Arabidopsis thaliana, provides a certain theoretical basis for establishing a complete plant regeneration and genetic transformation system for peony.

[0007] To achieve the above object, the present invention provides the following solutions:

[0008] The present invention provides an application of a peony PoLEC1 gene and a protein encoded by the gene in delaying bolting and flowering of Arabidopsis thaliana. The nucleotide sequence of the peony PoLEC1 gene is shown in SEQ ID NO.11; the amino acid sequence of the protein encoded by the peony PoLEC1 gene is shown in SEQ ID NO.12.

[0009] The invention provides a method for delaying bolting and flowering of Arabidopsis thaliana, comprising introducing a peony PoLEC1 gene into Arabidopsis thaliana to obtain a transgenic Arabidopsis thaliana plant stably expressing the peony PoLEC1 gene, and delaying bolting and flowering of the transgenic Arabidopsis thaliana plant; the nucleotide sequence of the peony PoLEC1 gene is shown in SEQ ID NO.11.

[0010] The present invention provides an application of a peony PoLEC1 gene and a protein encoded by the same in reducing the growth rate of Arabidopsis thaliana strains. The nucleotide sequence of the peony PoLEC1 gene is shown in SEQ ID NO.11; the amino acid sequence of the protein encoded by the peony PoLEC1 gene is shown in SEQ ID NO.12.

[0011] The present invention provides a method for reducing the plant height of Arabidopsis thaliana, comprising introducing a peony PoLEC1 gene into Arabidopsis thaliana to obtain a transgenic Arabidopsis thaliana plant stably expressing the peony PoLEC1 gene, so that the plant height of the transgenic Arabidopsis thaliana plant is reduced; the nucleotide sequence of the peony PoLEC1 gene is shown in SEQ ID NO.11.

[0012] The present invention provides an application of a peony PoLEC1 gene and a protein encoded by the gene in increasing the number of Arabidopsis leaves. The nucleotide sequence of the peony PoLEC1 gene is shown in SEQ ID NO.11; the amino acid sequence of the protein encoded by the peony PoLEC1 gene is shown in SEQ ID NO.12.

[0013] The present invention provides a method for increasing the number of Arabidopsis leaves, comprising introducing a peony PoLEC1 gene into Arabidopsis to obtain a transgenic Arabidopsis plant stably expressing the peony PoLEC1 gene, so that the number of leaves of the transgenic Arabidopsis plant is increased; the nucleotide sequence of the peony PoLEC1 gene is shown in SEQ ID NO.11.

[0014] The present invention provides an application of a peony PoLEC1 gene and a protein encoded by the gene in promoting Arabidopsis callus differentiation into buds. The nucleotide sequence of the peony PoLEC1 gene is shown in SEQ ID NO.11; the amino acid sequence of the protein encoded by the peony PoLEC1 gene is shown in SEQ ID NO.12.

[0015] The present invention provides a method for promoting Arabidopsis callus to differentiate into buds, comprising overexpressing a peony PoLEC1 gene in Arabidopsis to promote the Arabidopsis callus to differentiate into buds; the nucleotide sequence of the peony PoLEC1 gene is shown in SEQ ID NO.11.

[0016] The present invention discloses the following technical effects:

[0017] The present invention clones the peony PoLEC1 gene from the cultivated peony 'Fengdan', transforms it into Arabidopsis, and finds through phenotypic comparison that the peony PoLEC1 gene can play a role in plant bolting, flowering, growth and development, which is specifically reflected in that overexpression of the peony PoLEC1 gene can increase the number of leaves, delay bolting and flowering time, and reduce plant height. The present invention uses Arabidopsis as a model plant, explores the functional characteristics of the peony PoLEC1 gene and its expression characteristics in the entire somatic embryo direct generation process and in various tissues of overexpressed Arabidopsis, and provides a certain theoretical basis for establishing a complete plant regeneration and genetic transformation system for peony. BRIEF DESCRIPTION OF THE DRAWINGS

[0018] In order to more clearly illustrate the embodiments of the present invention or the technical solutions in the prior art, the drawings required for use in the embodiments will be briefly introduced below. Obviously, the drawings described below are only some embodiments of the present invention. For ordinary technicians in this field, other drawings can be obtained based on these drawings without paying creative work.

[0019] Figure 1 is the map of the overexpression plasmid pHG;

[0020] Figure 2 This is the PCR electrophoresis diagram of the tree peony PoLEC1 gene; M is the DL2000 maker; 1 is the target band;

[0021] Figure 3This is the phylogenetic tree of peony PoLEC1 protein and LEC1 of other species;

[0022] Figure 4 This is the amino acid sequence alignment of the peony PoLEC1 protein and LEC1 of other species; among them, the amino acid sequence similarity represented by dark blue is 100%, the amino acid sequence similarity represented by pink is ≥75%, and the amino acid sequence similarity represented by sky blue is ≥50%;

[0023] Figure 5 The wild-type tree peony (WT) and T 3 Phenotypes of transgenic Arabidopsis thaliana (OE-PoLEC1) overexpressing the gene; a is a picture taken at 20 days after planting; b is a picture taken at 28 days after planting; c is the number of leaves counted at 20 days after planting; d is the height of the plant counted at 40 days after planting; e is a picture taken at 40 days after planting; f and g are pictures taken at 15 days and 25 days after seeding, respectively;

[0024] Figure 6 The PoLEC1 gene in peony is expressed in T 3 Expression patterns in overexpressing transgenic Arabidopsis thaliana. DETAILED DESCRIPTION

[0025] Various exemplary embodiments of the present invention will now be described in detail. This detailed description should not be considered as limiting the present invention, but should be understood as a more detailed description of certain aspects, features, and embodiments of the present invention.

[0026] It should be understood that the terms described in the present invention are only for describing a particular embodiment and are not intended to limit the present invention. In addition, for the numerical range in the present invention, it should be understood that each intermediate value between the upper and lower limits of the scope is also specifically disclosed. The intermediate value in any stated value or stated range, and each smaller range between any other stated value or intermediate value in the described range is also included in the present invention. The upper and lower limits of these smaller ranges can be independently included or excluded in the scope.

[0027] Unless otherwise indicated, all technical and scientific terms used herein have the same meanings as those generally understood by those skilled in the art. Although the present invention describes only preferred methods and materials, any methods and materials similar or equivalent to those described herein may also be used in the implementation or testing of the present invention. All documents mentioned in this specification are incorporated by reference to disclose and describe the methods and / or materials associated with the documents. In the event of a conflict with any incorporated document, the content of this specification shall prevail.

[0028] It will be apparent to those skilled in the art that various modifications and variations may be made to the specific embodiments of the present invention description without departing from the scope or spirit of the present invention. Other embodiments derived from the present invention description will be apparent to those skilled in the art. The present invention description and examples are exemplary only.

[0029] The words “include,” “including,” “have,” “contain,” etc. used in this document are open-ended terms, meaning including but not limited to.

[0030] Example 1

[0031] 1. Experimental materials and methods

[0032] 1.1 Test Materials

[0033] The experimental material is the cultivar 'Fengdan', which has been cultivated and evolved for a long time and has ornamental, medicinal, oil-use characteristics and stable genetic traits. The seeds of 'Fengdan' that matured that year were selected, and the seeds came from Heze, Shandong. The peony tissue culture seedlings are developed from the embryo as explants. After 7 days of dark treatment, the explants are transferred to the tissue culture room for normal culture. The normal culture conditions in the tissue culture room are: alternating light for 16 hours and dark for 8 hours, and the light intensity is 40μmol·m -2 ·s -1 , temperature 24±1℃. Subculture once every 20 days. The roots, stems, leaves and callus of peony seeds and tissue culture seedlings were taken for RNA extraction for real-time fluorescence quantitative analysis. The Arabidopsis thaliana used in the present invention were Columbia-0 (Col-0) wild type, planted in the laboratory artificial climate room, with a light intensity of 40μmol·m -2 ·s -1 , humidity 60-70%, temperature 24±1℃.

[0034] 1.2 Methods

[0035] 1.2.1. Total RNA and cDNA synthesis

[0036] RNA was extracted from sterile peony tissue culture seedlings using a Quick RNA Isolation Kit (Huayueyang, China). The RNA concentration was determined by spectrophotometer. The integrity of the RNA band was confirmed by 1% agarose gel electrophoresis and the RNA was stored in a -80°C ultra-low temperature refrigerator for later use. The extracted RNA was used as a template and the TAKARA reverse transcription kit PrimeScript was used as a reference. TM The first strand of cDNA was synthesized by reverse transcription using the procedure of RT reagent Kit (Perfect Real Time) (RR047A).

[0037] 1.2.2 Cloning of the Paeonia suffruticosa PoLEC1 gene

[0038] According to the primer design principle, the primers were designed using SnapGene (V2.3.2) software (Table 1). The target sequence was amplified by using a high-fidelity enzyme (Vazyme, Nanjing, China) in Max Master Mix (P515). The target sequence was detected by 1% agarose gel electrophoresis. When the target band was consistent with the corresponding length of the marker sequence, it indicated that the target sequence was obtained and the CDS sequence of the peony PoLEC1 gene was cloned. The gel was cut on a UV gel cutting table, and the product was recovered according to the steps of the DNA purification and recovery kit (Tiangen, Beijing, China). The gel recovery product was connected to the cloning vector using the pCE2TA / Blunt-Zero vector (C601) (Vazyme, Nanjing, China) kit for transformation. Take Escherichia coli DH5α competent cells and transform them according to the steps. Then, evenly spread them on LB solid agar medium containing AMP and culture them upside down at 37°C for 14 hours. Then, select single clones for colony PCR, shake the positive single clones, extract the plasmid using TIANGEN Tiangen Plasmid Mini Extraction Kit (TIAN prep Mini Plasmid Kit, DP103-03, Beijing, China), and take 10 μL of the plasmid solution to send to Anshengda (Anshengda, Beijing, China) for sequencing to verify that the target sequence was obtained correctly.

[0039] Table 1 Detailed information of primers

[0040]

[0041] 1.2.3 Bioinformatics analysis

[0042] The basic physicochemical properties of the amino acid sequence of PoLEC1 were predicted and analyzed online using the ExPASy-ProtParam website. The secondary structure of the PoLEC1 protein was analyzed using SOPMA. TMHMM-v.2.0 and SignalP-v.6.0 were used to predict the presence or absence of transmembrane domains and signal peptides in PoLEC1, respectively. Plant mPLoc was used to predict the subcellular localization of the PoLEC1 protein. The PlantTFDB plant transcription factor database (https: / / planttfdb.gao-lab.org / ) and the NCBI public database (https: / / www.ncbi.nlm.nih.gov / ) were used to search for homology to the LEC1 protein sequences of other species. Clustal Wv.2.0.11 software was used to perform multiple sequence alignment of the amino acid sequence of peony LEC1 with the amino acid sequences of LEC1 of six species, including Arabidopsis thaliana, rapeseed, grape, corn, poplar, and cocoa. The phylogenetic tree was constructed by combining the LEC1 protein sequences of other species and the PoLEC1 protein sequences using the NJ (Neighbor-Joining) method of MEGA 11, with 1000 bootstrap replicates and other parameters set to default values. The tree was beautified online using iTOL (https: / / itol.embl.de / ).

[0043] DNAMAN 9.0 software was used to perform multiple sequence alignment of LEC1 protein sequences of peony, Arabidopsis, rapeseed, grape, cocoa, poplar and maize. MEME online software was used to perform motif analysis of PoLEC1 and LEC1 protein sequences of other species, and the number of motifs was set to 10. The saved XML file and the Newick file obtained in the evolutionary tree construction were input into the Gene Structure View (Advanced) window of TBtools v.2.4.0.119028 software for visualization and analysis.

[0044] 1.2.4 Functional verification of peony PoLEC1 gene transfection into Arabidopsis thaliana

[0045] The peony PoLEC1 gene was cloned into the plant expression vector pHG (pHG map as shown in Figure 1 The plant vector used in this example is pHG, and the 35S::PoLEC1 overexpressing transgenic Arabidopsis plants with stable inheritance were obtained by Agrobacterium-mediated inflorescence infection. 3 The overexpressing transgenic Arabidopsis thaliana and wild-type controls were planted and cultured under the same culture conditions, and observed and recorded.

[0046] 1.2.4.1. Observation of transgenic plant growth phenotype

[0047] Combine wild type and T 3 The seeds of the overexpressing transgenic Arabidopsis were sterilized and sown in a culture dish containing MS medium. After vernalization for 7 days, they were transplanted into planting soil and cultured in an artificial climate chamber. The number of leaves of Arabidopsis thaliana was counted at the same time after germination. The biological and technical replicates were repeated three times, and the growth was observed and photographed.

[0048] 1.2.4.2 Phenotypic observation of transgenic plant roots

[0049] Combine wild type and T 3 The seeds of the transgenic Arabidopsis thaliana overexpressing were sterilized and sown neatly side by side in a culture dish containing MS medium and cultured for 30 days after vernalization. The root development of Arabidopsis thaliana was observed and photographed.

[0050] 1.3 Fluorescence quantitative analysis

[0051] Seven stages of peony somatic embryo development (day 0 of peony somatic embryo development; day 5 of peony somatic embryo development; day 10 of peony somatic embryo development; day 15 of peony somatic embryo development; day 20 of peony somatic embryo development; day 30 of peony somatic embryo development; day 60 of peony somatic embryo development) and five different peony tissues (seed, root, stem, leaf and callus) were selected from the cultured peony in step 1.1. 3 The roots, stems, leaves, flowers, and fruits of the transgenic Arabidopsis thaliana were used to extract RNA and reverse transcribe the first strand of cDNA. 2 O was diluted 10 times as a template, and the primers are shown in Table 1. The reaction was completed on a QTOWER real-time fluorescence quantitative PCR instrument (analytikjena, Germany) using the TB Green Premix Ex TaqⅡ fluorescent quantitative kit (Tli RNaseH Plus, TaKaRa). The PCR reaction system was TB Green Premix Ex Taq 5μL; Template 1μL; Primer (F+R) 0.4μL; ddH 2 O was added to 10 μL, and a total of 3 biological replicates were set up. The reaction conditions were 95℃ pre-denaturation for 90s; 95℃ denaturation for 5s, 60℃ melting for 30s, 40 cycles; melting curve from 60 to 95℃, 1℃ increase every 15s. Peony ubiquitin ligase and AtActin were used as internal reference genes, and the expression levels of peony somatic embryos on day 0, peony root tissues, and Arabidopsis root tissues were used as control groups, respectively. The relative expression level was measured using 2 -ΔΔCTThe relative expression of PoLEC1 in various tissues was calculated and analyzed.

[0052] 2. Results and Analysis

[0053] 2.1 Cloning and sequence analysis of the tree peony PoLEC1 gene

[0054] Using the transcriptome data of somatic embryos from 10 different stages of peony as a reference, cDNA from the stage with higher expression of the target gene was selected as a template for gene cloning. The electrophoresis diagram showed that an obvious specific fragment appeared at the position of about 500 bp ( Figure 2 ), the length of the peony PoLEC1 gene is 582 bp, which is basically consistent with the expected target gene length. The target fragment is then connected to the cloning vector and sequenced to obtain 100% correct sequence information, and the desired target gene is cloned.

[0055] The nucleotide sequence of the peony PoLEC1 gene is shown in SEQ ID NO.11, specifically:

[0056] The basic physicochemical properties of the peony PoLEC1 gene were analyzed. The results showed that the peony PoLEC1 protein encoded by the gene had a total of 193 amino acids, a relative molecular weight of approximately 20.5 kDa, a theoretical isoelectric point of 5.98, an instability coefficient II of 29.10, an overall average hydrophilicity of -0.865, and a fat coefficient of 50.05. It is a stable hydrophilic protein (stability coefficient <40, hydrophilicity index is a negative value). Subcellular localization predicts that the LEC1 protein is localized in the cell nucleus.

[0057] The amino acid sequence of the peony PoLEC1 protein encoded by the peony PoLEC1 gene is shown in SEQ ID NO.12, specifically: MAESDDESGGHNTSGNANGELSQREQDRLLPIANVSRIMKKALPANAKISKDAKETVQECVSEFISFITGEASDKCQREKRKTINGDDLLWAMTTLGFEEYVEPLKVYLQKFREMEGEKTSVGRQGENSKDGSSGHGGSLVNSGNSGGGYGGGEMYGGMMLGNPHHPHPHGQMYGSGSYNHSGGGGASTGRPR*.

[0058] The secondary structure of the peony PoLEC1 protein is composed of a large number of α-helices and random coils, including 3 α-helices and 2 L-loops, and contains the highly conserved B domain in the HAP3 subunit, but no transmembrane domain and signal peptide.

[0059] 2.2 Phylogenetic tree analysis of peony PoLEC1 protein

[0060] To further understand the evolution and relationship between peony PoLEC1 protein and members of other species, the peony PoLEC1 protein was obtained and the LEC1 proteins of Arabidopsis, rapeseed, cocoa, grape, corn, soybean, kale, Jatropha curcas, and Selaginella were used to construct a phylogenetic tree ( Figure 3 The results showed that the PoLEC1 protein is closely related to cocoa and grapes.

[0061] 2.3 Multiple sequence comparison analysis of peony PoLEC1 protein

[0062] In order to analyze the similarity between peony PoLEC1 and LEC1 of other species, the amino acid sequence of peony PoLEC1 was compared with the LEC1 protein sequences of other closely related species. The results showed that the amino acid sequence of peony PoLEC1 protein contained three α-helical structures connected by two loops, which formed the B3 conserved domain of NF-YB subunit, and contained the conserved B region of HAP3 subunit ( Figure 4 ).

[0063] 2.4 Phenotypic analysis of Arabidopsis thaliana overexpressing the peony PoLEC1 gene

[0064] 2.4.1 Growth and development phenotypes of peony PoLEC1 transgenic plants

[0065] To explore the role of the peony PoLEC1 gene in plant growth and development, we observed the relationship between wild type and T 3 Phenotypic findings of the overexpressed transgenic Arabidopsis thaliana: 20 days after planting, T 3 There was no significant difference in leaf size between the overexpressing transgenic Arabidopsis and the wild type, and the number of leaves was slightly more than that of the wild type ( Figure 5 a and c); T 3 The bolting of the transgenic Arabidopsis thaliana overexpressing α-glucose was later than that of the wild type ( Figure 5 b); continue to observe and cultivate, and count the heights of the two plants after about 40 days of cultivation, and find that T 3 The plants overexpressing the transgenic Arabidopsis thaliana were shorter than the wild type ( Figure 5 d and e in the figure). 3 The transgenic Arabidopsis seeds and wild-type Arabidopsis seeds were sterilized and sown on MS medium containing hygromycin. The root growth status was observed and it was found that there was no significant difference in the length and density of the roots between the two. Figure 5 f and g in the figure). The above results indicate that the peony PoLEC1 gene can play a role in plant bolting, flowering, growth and development. Specifically, overexpression of the peony PoLEC1 gene can increase the number of leaves, delay the time of bolting and flowering, and reduce the height of the plant.

[0066] 2.5 Expression pattern analysis

[0067] The RT-qPCR method was used to 3 The expression levels of the peony PoLEC1 gene in the transgenic Arabidopsis thaliana overexpressing gene in various tissues of Arabidopsis thaliana: root, stem, leaf, flower, and fruit were analyzed ( Figure 6 ).

[0068] In T 3 When the transgenic Arabidopsis plants were cultured for about 45 days, their roots, stems, leaves, flowers and fruit pods were taken to extract RNA and reverse transcribe into cDNA, and the peony PoLEC1 gene was quantified in the roots, stems, leaves, flowers and fruit pods. Three biological replicates and three technical replicates were performed for each tissue. The results showed that the PoLEC1 gene was expressed in all five tissues of transgenic Arabidopsis, with the highest expression in leaves. This is consistent with the expression pattern of the PoLEC1 gene in peony, and it may play a role in the leaf development process.

[0069] The embodiments described above are only descriptions of the preferred modes of the present invention, and are not intended to limit the scope of the present invention. Without departing from the design spirit of the present invention, various modifications and improvements made to the technical solutions of the present invention by ordinary technicians in this field should all fall within the protection scope determined by the claims of the present invention.

Claims

1. Application of the peony PoLEC1 gene and the protein encoded therein in delaying bolting and flowering of Arabidopsis thaliana, characterized in that: The nucleotide sequence of the peony PoLEC1 gene is shown in SEQ ID NO.11; the amino acid sequence of the protein encoded by the peony PoLEC1 gene is shown in SEQ ID NO.

12.

2. A method for delaying bolting and flowering of Arabidopsis thaliana, characterized in that: The method comprises introducing the peony PoLEC1 gene into Arabidopsis thaliana to obtain a transgenic Arabidopsis thaliana plant stably expressing the peony PoLEC1 gene, so that the transgenic Arabidopsis thaliana plant delays bolting and flowering; the nucleotide sequence of the peony PoLEC1 gene is shown in SEQ ID NO.

11.

3. Application of the peony PoLEC1 gene and the protein encoded by it in reducing the high yield of Arabidopsis thaliana strains, characterized in that: The nucleotide sequence of the peony PoLEC1 gene is shown in SEQ ID NO.11; the amino acid sequence of the protein encoded by the peony PoLEC1 gene is shown in SEQ ID NO.

12.

4. A method for reducing the plant height of Arabidopsis thaliana, characterized in that: The method comprises introducing the peony PoLEC1 gene into Arabidopsis thaliana to obtain a transgenic Arabidopsis thaliana plant stably expressing the peony PoLEC1 gene, so that the plant height of the transgenic Arabidopsis thaliana plant is reduced; the nucleotide sequence of the peony PoLEC1 gene is shown in SEQ ID NO.

11.

5. Use of the peony PoLEC1 gene and the protein encoded therein for increasing the number of Arabidopsis leaves, characterized in that: The nucleotide sequence of the peony PoLEC1 gene is shown in SEQ ID NO.11; the amino acid sequence of the protein encoded by the peony PoLEC1 gene is shown in SEQ ID NO.

12.

6. A method for increasing the number of Arabidopsis leaves, characterized in that: The method comprises introducing the peony PoLEC1 gene into Arabidopsis thaliana to obtain a transgenic Arabidopsis thaliana plant stably expressing the peony PoLEC1 gene, so that the number of leaves of the transgenic Arabidopsis thaliana plant is increased; the nucleotide sequence of the peony PoLEC1 gene is shown in SEQ ID NO.

11.

7. Application of the peony PoLEC1 gene and the protein encoded by it in promoting the differentiation of Arabidopsis callus into buds, characterized in that: The nucleotide sequence of the peony PoLEC1 gene is shown in SEQ ID NO.11; the amino acid sequence of the protein encoded by the peony PoLEC1 gene is shown in SEQ ID NO.

12.

8. A method for promoting Arabidopsis callus differentiation into buds, characterized in that: The method comprises overexpressing the peony PoLEC1 gene in Arabidopsis thaliana to promote the differentiation of the Arabidopsis thaliana callus into buds; the nucleotide sequence of the peony PoLEC1 gene is shown in SEQ ID NO.11.

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