PfC3H71 gene of perilla frutescens and application of PfC3H71 gene in aspect of improving flowering of plants

Abstract

The invention discloses an application of a purple perilla flowering related gene PfC3H71 in regulation and control of plant flowering performance. The nucleotide sequence of the PfC3H71 gene is as shown in SEQ ID NO: 1. The recombinant vector pCAMBIA1303-PfC3H71 comprises a nucleotide sequence as shown in SEQ ID NO: 1 (Sequence Identity Number 1). The PfC3H71 gene of the C3H family of perilla frutescens is researched, so that the flowering mechanism of perilla frutescens can be understood, and a gene engineering technology and the like are utilized to induce plants to flower in advance, so that a theoretical basis and a reference are provided for mechanism research on regulation and control of a growth period and a flowering period.

Description

Technical Field

[0001] This invention belongs to the field of biogenetics technology, specifically relating to the perilla PfC3H71 gene and its application in improving plant flowering. Background Technology

[0002] Perilla frustescens (L.) Britt., an annual herb belonging to the genus Perilla in the family Lamiaceae, is a strictly short-day plant, sensitive to photoperiod. Perilla seeds are rich in polyunsaturated fatty acids, especially α-linolenic acid (ALA), which accounts for over 60%. Perilla is rich in polyphenols, carotenoids, flavonoids, anthocyanins, and other bioactive substances and nutrients, possessing significant application and development value. Due to the relatively recent development of perilla genomics, current research focuses on a few agronomic traits, including yield, perilla oil, leaf color, and seed germination, with limited research on perilla gene function. Currently, research on perilla gene function mainly focuses on genes related to oil synthesis and monoterpene biosynthesis, with limited research on the mechanisms of perilla flower development, and even fewer reports on genes involved in the biological process of flowering transition.

[0003] Flowering is a crucial stage in the life cycle of angiosperms, marking the transition from vegetative to reproductive growth. Flower formation is subject to a complex arrangement and strict regulation of multiple signaling networks, including the autonomous pathway, vernalization pathway, gibberellin pathway, photoperiod pathway, ambient temperature pathway, and age pathway. These six pathways converge and regulate flowering integration genes, such as FLOWERING LOCUS T (FT), TWIN SISTEROF FT (TSF), and TERMINAL FLOWER1 (TFL1), thereby regulating the transformation of shoot apical meristem into inflorescence meristem and inducing flowering.

[0004] The zinc finger transcription factor family is one of the largest transcription factor families in plants, playing a crucial role in plant morphogenesis, signal transduction, and responses to environmental stresses. Zinc finger transcription factors contain zinc finger motifs, three-dimensional "finger-like" structures formed by cysteine ​​(C) and histidine (H) residues arranged in a specific spacing pattern, and stabilized by zinc ions. Based on their structural diversity, zinc finger transcription factors can be classified into C2H2, C8, C6, C3HC4, C2HC, C2HC5, C4, C4HC3, and C3H. The C3H zinc finger gene family has been analyzed genome-wide in Arabidopsis thaliana, rice, maize, soybean, poplar, tomato, and tobacco, identifying 68, 67, 68, 116, 91, 97, and 86 C3H zinc finger genes, respectively. In plants, C3H zinc finger proteins are involved in plant growth and development, abiotic and biotic stress responses, and plant hormone regulation. However, to date, there has been no systematic identification and analysis of the members and functions of the C3H zinc finger gene family in perilla. Therefore, there is significant room for exploration in the systematic analysis and functional study of the C3H gene family in perilla. Summary of the Invention

[0005] The purpose of this invention is to provide the perilla PfC3H71 gene and its application in improving plant flowering.

[0006] The PfC3H71 gene of Perilla frutescens, wherein the polynucleotides of the PfC3H71 gene are shown in (a), (b), (c), or (d):

[0007] (a) a polynucleotide as shown in SEQ ID No: 1; or

[0008] (b) A polynucleotide whose complementary sequence to SEQ ID No: 1 can hybridize under strict hybridization conditions, and the protein encoded by the polynucleotide still has the function of enhancing plant flowering;

[0009] (c) A polynucleotide having at least 90% or more homology with the polynucleotide shown in SEQ ID No: 1; or

[0010] (d) A polynucleotide mutant obtained by deleting, substituting or inserting one or more bases based on the polynucleotide shown in SEQ ID No: 1, wherein the protein encoded by the polynucleotide mutant still has the activity of improving plant flowering.

[0011] The perilla PfC3H71 protein, wherein the amino acid sequence of the PfC3H71 protein is shown in (a), (b), or (c):

[0012] (a) The amino acid sequence as shown in SEQ ID No: 2 of the sequence listing; or

[0013] (b) Amino acids that are at least 90% homologous to the amino acid shown in SEQ ID No: 2; or

[0014] (c) A protein mutant obtained by deleting, substituting or inserting one or more amino acids based on the protein shown in SEQ ID No: 2, and the protein still has the activity of improving plant flowering.

[0015] A plasmid containing the PfC3H71 gene, wherein the plasmid containing the PfC3H71 gene is obtained by ligating the PfC3H71 gene with the plasmid vector pMD19-T.

[0016] The PfC3H71 gene editing recombinant vector is obtained by using the plasmid containing the PfC3H71 gene, obtaining the PfC3H71 gene sequence containing a portion of the vector sequence through PCR, and ligating it into the pCAMBIA1303 vector plasmid after Spe I single enzyme digestion reaction, resulting in a PfC3H71 gene editing recombinant vector driven by the 35S promoter.

[0017] Agrobacterium competent cells containing the PfC3H71 gene, wherein the Agrobacterium competent cells containing the PfC3H71 gene are obtained by introducing the PfC3H71 gene editing recombinant vector into Agrobacterium.

[0018] Application of the PfC3H71 gene in promoting plant flowering.

[0019] The application involves using Agrobacterium tumefaciens competent cells containing the PfC3H71 gene to transform Arabidopsis thaliana, thereby obtaining transformants and enabling the transformants to flower earlier.

[0020] The beneficial effects of this invention: By studying the PfC3H71 gene of the Perilla C3H family, this invention helps to understand the flowering mechanism of Perilla and uses genetic engineering technology to induce plants to flower earlier, thus providing a theoretical basis and reference for the study of the mechanism of regulating growth period and flowering period. Attached Figure Description

[0021] Figure 1 Tissue-specific expression analysis of PfC3H71 in Perilla frutescens.

[0022] Figure 2 This is a gel electrophoresis image of the PfC3H71 gene clone; M represents DL 5000 Marker, and 1 represents the target fragment.

[0023] Figure 3 This is a phylogenetic tree diagram, where Pf represents perilla, Os represents rice, At represents Arabidopsis thaliana, and all C3H genes are divided into ten groups (I-X).

[0024] Figure 4 Electrophoresis diagram for PCR molecular identification of transgenic Arabidopsis thaliana.

[0025] Figure 5 The image shows the growth status of transgenic Arabidopsis thaliana under LD conditions after 25 days; Col-0 and OC-1, OC-2, and OC-3 represent wild-type Arabidopsis thaliana and transgenic Arabidopsis thaliana with the PfC3H71 gene, respectively, and the white triangles represent flowering sites.

[0026] Figure 6 The flowering time typology of wild-type and transgenic Arabidopsis thaliana under LD conditions is shown. Asterisks indicate significant differences in Dunnett's test after one-way ANOVA, and * and ** represent P<0.05 and P<0.01, respectively.

[0027] Figure 7 The plant height of wild-type and transgenic Arabidopsis thaliana lines at 45 days under LD conditions is given. The asterisk indicates a significant difference in the Dunnett test after one-way ANOVA. * and ** represent P<0.05 and P<0.01, respectively.

[0028] Figure 8 The image shows the growth status of transgenic Arabidopsis thaliana under SD conditions after 45 days; Col-0 and OC-1, OC-2, and OC-3 represent wild-type Arabidopsis thaliana and transgenic Arabidopsis thaliana with the PfC3H71 gene, respectively; white triangles represent flowering sites; and white rhombuses represent silique development sites.

[0029] Figure 9 The flowering time typology of wild-type and transgenic Arabidopsis thaliana under SD conditions is shown. Asterisks indicate significant differences in Dunnett's test after one-way ANOVA, and * and ** represent P<0.05 and P<0.01, respectively.

[0030] Figure 10 The plant height of wild-type and transgenic Arabidopsis thaliana lines at 65 days under SD conditions is given by ns, where ns represents the Dunnett test result after one-way ANOVA, where P > 0.05.

[0031] Figure 11 The silique development status of wild-type Arabidopsis and transgenic Arabidopsis under SD conditions at 65 days. Detailed Implementation

[0032] To facilitate understanding of the present invention, a more comprehensive description will be given below. However, the present invention can be implemented in many different forms and is not limited to the embodiments described herein. Rather, these embodiments are provided to provide a thorough and complete understanding of the disclosure of the present invention.

[0033] Example 1

[0034] 1. RNA extraction and real-time quantitative PCR analysis from various tissues of Perilla frutescens

[0035] RNA was extracted from the roots, stems, leaves, flowers, and mature seeds of WT and EM-type Perilla frutescens grown under LD and SD conditions using the Tiangen RNA prep Pure plant total RNA extraction kit, following its instructions. RNA content was detected using a Nano 100C instrument from Hangzhou Aosheng Company, and integrity was assessed using 1% agarose gel electrophoresis.

[0036] Strictly in accordance with TaKaRa's PrimeScript TM The instructions for the RT reagent kit with gDNAEraser (Perfect Real Time) show that perilla RNA was reverse transcribed to obtain cDNA. The resulting cDNA was stored at -20°C for later use.

[0037] By reviewing the transcriptome data of *Perilla frutescens*, quantitative real-time PCR primers (PfC3H71-qF and PfC3H71-qR) were designed using Primer 5.0 software. The relative expression level of PfC3H71 was analyzed using Bio-Rad's SsoAdvanced Universal SYBR Green Supermix. *Perilla frutescens* PfActin was used as an internal reference gene, and quantitative analysis was performed on a CFX96 Real-Time PCR Detection System (Bio-Rad, USA). The real-time quantitative PCR (RT-qPCR) reaction was as follows: 95℃ for 30 s, 95℃ for 10 s, 60℃ for 20 s, for 40 cycles.

[0038] The primer sequence used for quantitative fluorescence verification of PfC3H71 is as follows:

[0039] PfC3H71-qF:5'-CTACAAATAGGCAGGTGACG-3' (SEQ ID NO: 3);

[0040] PfC3H71-qR: 5'-AAGTTTTCCAGTGGGAGAGA-3' (SEQ ID NO: 4).

[0041] The primer sequences used for the internal reference gene PfActin are as follows:

[0042] Pf-β-Actin-F: 5'-AGACCTTCAATGTGCCAGCCA-3' (SEQ ID NO: 5);

[0043] Pf-β-Actin-R: 5'-CACGACCAGCAAGATCCAACC-3' (SEQ ID NO: 6).

[0044] Use 2 -△△CT The relative expression levels of the PfC3H71 gene in different tissues were calculated using the method described above. The results are shown in [Figure 1]. Figure 1 Under LD conditions, PfC3H71 was expressed in roots, stems, leaves, flowers, and seeds. The highest expression level was observed in the roots of WT-type perilla and in the leaves of EM-type perilla. Under SD conditions, the highest expression level was observed in the leaves of WT-type perilla and in the roots of EM-type perilla.

[0045] 2. Gene cloning

[0046] By reviewing the perilla transcriptome data, cloning primers (PfC3H71-F and PfC3H71-R) targeting the PfC3H71 gene were designed. Using perilla seed cDNA as a template, PfC3H71 was cloned via RT-PCR. The polymerase used for PfC3H71 gene cloning was Phanta Super-Fidelity DNA Polymerase from Nanjing Vazyme. The reaction system was as follows: 1.0 μL cDNA (template), 6 μL Premix Taq (containing dNTAs and Mg2+), 1.0 μL forward and reverse primers (10 μM), 16 μL sterile water, total volume 25 μL. The RT-PCR reaction was performed as follows: 30 cycles of 95℃ for 1 min, 95℃ for 10 s, 60℃ for 20 s, and 72℃ for 50 s, followed by 72℃ for 10 min and 4℃ for 9 min.

[0047] The polymerase used for cloning was Taq polymerase from TaKaRa. TM Version 2.0, the reaction system is as follows: 2 μL cDNA (template), 25 μL Premix Taq (containing dNTAs and Mg2+), 2 μL forward and reverse primers (10 μM), 19 μL sterile water, total volume 50 μL. The RT-PCR reaction is as follows: 95℃ for 1 min, 95℃ for 15 s, 58℃ for 15 s, 72℃ for 90 s for 32 cycles, 72℃ for 5 min, 4℃ for 10 min.

[0048] The primers used for RT-PCR amplification of the full-length PfC3H71 gene are:

[0049] PfC3H71-F:5'-ATGATGGTTGGAGAGAGAAGCT-3' (SEQ ID NO: 7);

[0050] PfC3H71-R: 5'-TCACTTGATCAGATCCGACACC-3' (SEQ ID NO: 8).

[0051] PCR products were detected by 1% agarose gel electrophoresis, and the target fragment was then recovered and purified using the SanPrep column-based DNA gel extraction kit from Shanghai Sangon Biotech. The purified product was ligated into the pMD19-T vector and transformed into competent *E. coli* DH5α. The transformed *E. coli* were cultured on IXA solid medium (containing Amp, X-gal, and IPTG) for 12-15 h. White single colonies were picked and cultured on LB liquid medium (containing 50 mg / mL Amp) on a shaker for 12 h. After verification by colony PCR, the colonies were sent to Shanghai Sangon Biotech for sequencing. Results are as follows: Figure 2 As shown, the PfC3H71 nucleic acid sequence, as shown in SEQ ID NO: 1, can encode an amino acid sequence of length 338, as shown in SEQ ID NO: 2.

[0052] Sequences of C3H proteins from Arabidopsis thaliana and rice (Oryza sativa L.) were downloaded from the TAIR and NCBI databases, respectively. The pairwise distance matrix was estimated using the JTT model, and a heuristic initial search tree was constructed using the Neighbor-Join algorithm, selecting topologies with high log-likelihood values. A phylogenetic tree of the aforementioned C3H proteins was then constructed using the adjacency-joining method in MEGA 11 software. The results are as follows: Figure 3 As shown, the protein encoded by the PfC3H71 gene of Perilla frutescens is closely related to AtC3H49 and AtC3H20 of Arabidopsis thaliana.

[0053] Example 2: Construction of the pCAMBIA1303 recombinant vector

[0054] Primers were designed using the full-length cDNA sequence of PfC3H71 as a template. Restriction sites and protective bases were added to both ends of the pCAMBIA1303 cDNA by alignment with the pCAMBIA1303 restriction site. The primer sequences are as follows:

[0055] PfC3H71-IF-F:

[0056] 5'-TTGACCATGGTAGATCTGACTAGTGTTGGAGAGAAGCTATGGCGGG-3' (SEQ ID NO: 9);

[0057] PfC3H71-IF-R:

[0058] 5'-GGTTTCTACAGGACGTAAACTAGTCTTGATCAGATCCGACACCCAACC-3' (SEQ ID NO: 10).

[0059] Colony PCR was performed on pMD19-T-PfC3H71 bacterial culture. After electrophoresis, the culture was purified and recovered using a gel extraction kit to obtain the PfC3H71 gene sequence containing a portion of the vector sequence. pCAMBIA1303 was digested with a single enzyme in a 50 μL volume: 5 μL 10X QuickCut Buffer, 5 μL plasmid DNA, 1 μL Spe I, and 39 μL ddH2O. The reaction was carried out at 37°C for 5 min. Then, the PfC3H71 gene with restriction sites and protective bases was ligated to the digested linear pCAMBIA1303 vector. The ligation mixture consisted of 2 μL linear pCAMBIA1303 plasmid, 3 μL template DNA, 0.2 μL In-Fusion enzyme, and 4.8 μL sterile water. The ligation product was transformed into competent E. coli DH5α, plated on kanamycin-containing resistant plates, and single colonies were picked and sequenced. After confirming the sequence, the plasmid pCAMBIA1303-PfC3H71 was extracted. The identification primers selected were MI-F and MI-R, MI-F (5'-TTTCATTTGGAGAGAACACGGGGGA-3'SEQ ID NO: 11) and MI-R (5'-CGCTGATCAATTCCACAGTTTTCGC-3'SEQ ID NO: 12).

[0060] The constructed plant expression vector pCAMBIA1303-PfC3H71 was transformed into Agrobacterium GV3101 using the freeze-thaw method.

[0061] Example 3: Transgenic Arabidopsis obtained by Agrobacterium transformation.

[0062] 1. Plant expression vector integrated into the Arabidopsis genome

[0063] Arabidopsis seeds were added to sterile water and placed in a refrigerator at 4°C for 24 hours. They were then disinfected with a 2% sodium hypochlorite solution for 1 minute, rinsed, and evenly sown on the surface of 1 / 2 MS solid medium. The seeds were then cultured at 22°C under long-day conditions (light / dark cycle of 16 / 8 hours) for 10 days. When the Arabidopsis seedlings had four true leaves, they were transplanted into a composite soil mixture of vermiculite, nutrient soil, and fine soil in a 1:2:4 ratio. Inflorescence inoculation was performed when the Arabidopsis reached the bolting stage.

[0064] Agrobacterium carrying the target plasmid was transferred to fresh LB medium (containing the same antibiotic and 150 μM AS) at a ratio of 1:50 and cultured at 28°C with shaking at 200 rpm until OD600 ≈ 0.8–1.0. The culture was then centrifuged at 4000 rpm for 10 minutes. After centrifugation, the supernatant was discarded, and the bacterial pellet was collected. 20 mL of infection solution (5 g Sucrose, 10 μL Silwet L-77, and ddH2O to a final volume of 100 mL) was added to the pellet, and the suspension was thoroughly mixed by pipetting with a disposable dropper.

[0065] Select Arabidopsis plants with well-developed main inflorescences and lateral branches, and whose flowers have not yet withered. Remove the fully opened flowers and pods, leaving only the flower buds. Invert the plants so that the flower buds are completely immersed in a suspension containing Agrobacterium for 2 minutes, continuously agitating the surface of the liquid to promote bacterial movement. After infection, remove the plants and provide ample water, then cover them with a transparent cover to maintain humidity and incubate in the dark for 24 hours. Remove the cover and return the plants to normal light conditions for further cultivation. To improve transformation efficiency, a second infection can be performed, with a 7-day interval between each infection.

[0066] Mature seeds of Arabidopsis thaliana after infection were harvested. Positive plants (T1 generation) with normal rooting and green leaves were selected using 1 / 2 MS + Hyg medium and transplanted into multi-layered soil for further cultivation. The processes of 1 / 2 MS + Hyg resistance plate selection, PCR molecular identification, and GUS staining were repeated until homozygous T3 generation plants that did not exhibit phenotypic segregation were harvested.

[0067] 2. Identification of transgenic plants overexpressing PfC3H71

[0068] gDNA was extracted from young leaves of T0 generation Arabidopsis thaliana transgenic with the PfC3H71 gene. gDNA from wild-type Arabidopsis thaliana was used as a negative control for colony PCR verification. PCR amplification was performed using PfC3H71-IF-F and PfC3H71-IF-R, MI-F, and MI-R as primers, respectively. Figure 4 The results showed that all transgenic Arabidopsis thaliana strains were successfully amplified, indicating that the overexpression of PfC3H71 Arabidopsis thaliana was successfully transformed. Then, after three rounds of 1 / 2 MS+Hyg resistance plate screening and molecular identification, T3 homozygous lines were obtained. We obtained three T3 generation homozygous transgenic positive seedlings, named OC-1, OC-2, and OC-3, respectively. Subsequent experiments all used T3 homozygous seedlings as material.

[0069] Example 4: Phenotypic Analysis of Arabidopsis thaliana T3-transformed with PfC3H71

[0070] 1. Phenotypic analysis of Arabidopsis thaliana PfC3H71 under LD conditions

[0071] Three homozygous transgenic Arabidopsis thaliana lines (OC-1, OC-2, and OC-3) from the T3 generation were selected for further observation. Wild-type Col-0 was used as a control. All lines were cultured simultaneously under LD conditions, and the opening time of the first flower was observed. Phenotypic results are as follows: Figure 5 The transgenic lines flowered significantly earlier than the wild type. Phenotypic statistics are as follows: Figure 6 The flowering times of OC-1, OC-2, and OC-3 were 26.3±1.1 days, 26.3±0.8 days, and 26.5±0.9 days, respectively; the flowering time of wild type Col-0 was 31.7±1.1 days, which was 5.3 days earlier than that of wild type.

[0072] Meanwhile, PfC3H71 overexpression enhances the vegetative growth advantage of Arabidopsis thaliana, as shown in the phenotypic statistics. Figure 7 The plant height of the transgenic lines at 45 days (OC-1: 50.2±1.9cm; OC-2: 52.7±3.1cm; OC-3: 51.6±2.1cm) was significantly increased compared to the wild type (47.5±1.9cm). This result reveals that PfC3H71 has the dual function of accelerating flowering transition and promoting vegetative growth.

[0073] 2. Phenotypic analysis of Arabidopsis thaliana PfC3H71 under SD conditions

[0074] Wild-type Col-0 and T3 generation overexpression lines (OC-1, OC-2, OC-3) were placed together under SD conditions for phenotypic observation and analysis. Figure 8 Short-day conditions significantly delayed the flowering process in Arabidopsis thaliana, but transgenic plants flowered significantly earlier than wild-type plants. Phenotypic statistics are as follows: Figure 9 The flowering time of wild-type Arabidopsis thaliana was 50.3±0.4 days, while the flowering times of the transgenic lines OC-1, OC-2, and OC-3 were 37±0.4 days, 37.1±0.3 days, and 37±0.3 days, respectively, approximately 13 days earlier than the wild type. Furthermore, the plant height of wild-type Arabidopsis thaliana at 65 days was 55.8±1 cm, while the plant heights of the OC-1, OC-2, and OC-3 transgenic Arabidopsis thaliana lines at 65 days were 56.2±0.8 cm, 56.1±0.6 cm, and 55.8±1 cm, respectively. Overexpression of PfC3H71 had no effect on plant height under short-day conditions. Figure 10 The developmental state of the silique is as follows: Figure 11 Although short-day conditions restrict the development of siliques in the whole plant, the transgenic lines have better silique development than the wild type, and their morphology is closer to the normal fruiting phenotype under long-day conditions.

[0075] The embodiments described above are merely illustrative of several implementations of the present invention, and while the descriptions are relatively specific and detailed, they should not be construed as limiting the scope of the invention patent. It should be noted that those skilled in the art can make various modifications and improvements without departing from the concept of the present invention, and these all fall within the protection scope of the present invention. Therefore, the protection scope of this invention patent should be determined by the appended claims.

Claims

1. The Perilla PfC3H71 gene, characterized by, The polynucleotides of the PfC3H71 gene are shown in (a), (b), (c), or (d): (a) a polynucleotide as shown in SEQ ID No: 1; or (b) A polynucleotide whose complementary sequence to SEQ ID No: 1 can hybridize under strict hybridization conditions, and the protein encoded by the polynucleotide still has the function of enhancing plant flowering; (c) A polynucleotide having at least 90% or more homology with the polynucleotide shown in SEQ ID No: 1; or (d) A polynucleotide mutant obtained by deleting, substituting or inserting one or more bases based on the polynucleotide shown in SEQ ID No: 1, wherein the protein encoded by the polynucleotide mutant still has the activity of improving plant flowering.

2. Perilla PfC3H71 protein, characterized in that, The amino acid sequence of the PfC3H71 protein is shown in (a), (b), or (c): (a) The amino acid sequence as shown in SEQ ID No: 2 of the sequence listing; or (b) Amino acids that are at least 90% homologous to the amino acid shown in SEQ ID No: 2; or (c) A protein mutant obtained by deleting, substituting or inserting one or more amino acids based on the protein shown in SEQ ID No: 2, and the protein still has the activity of improving plant flowering.

3. A plasmid containing the PfC3H71 gene, characterized in that, The plasmid containing the PfC3H71 gene is obtained by ligating the PfC3H71 gene as described in claim 1 with the plasmid vector pMD19-T.

4. A PfC3H71 gene editing recombinant vector, characterized in that, The PfC3H71 gene editing recombinant vector is obtained by using the plasmid containing the PfC3H71 gene as described in claim 3, obtaining the PfC3H71 gene sequence containing a portion of the vector sequence by PCR, and ligating it into the pCAMBIA1303 vector plasmid after Spe I single enzyme digestion reaction, resulting in a PfC3H71 gene editing recombinant vector driven by the 35S promoter.

5. Agrobacterium competent cells containing the PfC3H71 gene, characterized in that, The Agrobacterium competent cells containing the PfC3H71 gene were obtained by introducing the PfC3H71 gene editing recombinant vector as described in claim 4 into Agrobacterium.

6. Application of PfC3H71 gene in promoting plant flowering period.

7. The application according to claim 6, characterized in that, The application involves using Agrobacterium tumefaciens competent cells containing the PfC3H71 gene to transform Arabidopsis thaliana, thereby obtaining transformants and enabling the transformants to flower earlier.

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