CeNF-YA1 gene, expression vector and application of CeNF-YA1 gene in regulation and control of vegetable fat

Abstract

The invention belongs to the technical field of molecular biology, and particularly relates to a CeNF-YA1 gene, an expression vector and application of the CeNF-YA1 gene in vegetable fat regulation and control. The invention provides the CeNF-YA1 gene for the first time, the nucleotide sequence of the CeNF-YA1 gene is as shown in SEQ ID NO.1, the full length of a coding region of the CeNF-YA1 gene is 792 bp, a series of carriers containing the CeNF-YA1 gene, such as subcellular localization, yeast hybridization and plant overexpression, are constructed for the first time, and meanwhile, the CeNF-YA1 protein is proved to have a transcription activation function, is localized in a cell nucleus and conforms to the basic characteristics of transcription factors. It is found for the first time that overexpression of the CeNF-YA1 gene can significantly improve the grease content of arabidopsis seeds and leaves; therefore, the technical scheme provided by the invention can be used for improving the oil content of the plant seeds and the nutritional tissues, and shows the application prospect in improving the oil content of the seeds and the nutritional tissues.

Description

Technical Field

[0001] This invention belongs to the field of molecular biology technology, specifically involving CeNF-YA1 Genes, expression vectors and their applications in the regulation of plant oils. Background Technology

[0002] Tiger nuts ( Cyperus esculentus (L.) belongs to the Cyperaceae family of the Poales order. Also known as oil sedge or tiger nut, it is a root crop with high nutritional value. It is characterized by its wide adaptability, great development potential, and high oil yield per acre, making it a highly sought-after new food ingredient in the fields of health food and sustainable agriculture.

[0003] Dietary lipid synthesis and accumulation are controlled by a multi-gene regulatory network, in which transcription factors play a crucial role, such as AP2 / EREBP, B3, NF-Y, MYB, WRKY, DOF, and bZIP. WRI1 ( WRINKLED1 This gene belongs to the AP2 / ERF transcription factor family and is a key gene confirmed to regulate lipid accumulation during oilseed seed development. Currently, information regarding tiger nuts... WRI Gene-like studies have been reported, for example, in Chinese patent CN 118531058 A, on a tiger nut. CeWRI1b The application of genes in the regulation of plant oils provides CeWRI1b Genes, and confirmed CeWRI1b Overexpression can be complementary atwri1-1 The wrinkled phenotype and oil content of mutant seeds can significantly increase the oil content of leaves; another example is Chinese patent CN118064452 A, a tiger nut. CeWRI2 Genes, expression vectors, and their applications in plant oil regulation have led to the discovery of... CeWRI2 Genes can be used to increase the oil content of plant seeds and vegetative tissues; Chinese Patent CN 118064455 A: A Kind of Tiger Nut CeWRI3 Genes, expression vectors, and their applications in plant oil regulation have led to the discovery of... CeWRI3 Genes can be used to increase the oil content of plant seeds and vegetative tissues. However, no such gene has been observed to date. CeNF-YA1 Reports on genes and their role in increasing the oil content of plant seeds and vegetative tissues. Summary of the Invention

[0004] The purpose of this invention is to provide CeNF-YA1 Genes, expression vectors and their applications in the regulation of plant oils.

[0005] This invention provides CeNF-YA1 Genes, the ones mentioned CeNF-YA1The nucleotide sequence of the gene is shown as SEQ ID NO. 1.

[0006] The present application provides the above technical solution CeNF-YA1 The CeNF-YA1 protein encoded by the gene, and the amino acid sequence of the CeNF-YA1 protein is shown as SEQ ID NO. 11.

[0007] The present application provides the above technical solution CeNF-YA1 The expression vector of the gene, and the expression vector comprises one or more of the following: a cell positioning vector pNC-Cam1304-CeNF-YA1, a yeast hybridization vector, and a plant overexpression vector pCAMBIA1301-CeNF-YA1. The yeast hybridization vector is one or more of the following: pNC-GBKT7-CeNF-YA1F, pNC-GBKT7-CeNF-YA1N1, pNC-GBKT7-CeNF-YA1N2, and pNC-GBKT7-CeNF-YA1C.

[0008] Preferably, the pNC-Cam1304-CeNF-YA1 is constructed by CeNF-YA1F2 and CeNF-YA1R2; Or the pNC-GBKT7-CeNF-YA1F is constructed by CeNF-YA1F2 and CeNF-YA1R2; The pNC-GBKT7-CeNF-YA1N1 is constructed by CeNF-YA1F2 and CeNF-YA1R3; The pNC-GBKT7-CeNF-YA1N2 is constructed by CeNF-YA1F2 and CeNF-YA1R4; The pNC-GBKT7-CeNF-YA1C is constructed by CeNF-YA1F3 and CeNF-YA1R2; The pCAMBIA1301-CeNF-YA1 is constructed by CeNF-YA1F4 and CeNF-YA1R5; The nucleotide sequence of the CeNF-YA1F2 is shown as SEQ ID NO. 4; The nucleotide sequence of the CeNF-YA1R2 is shown as SEQ ID NO. 5; The nucleotide sequence of the CeNF-YA1F3 is shown as SEQ ID NO. 7; The nucleotide sequence of the CeNF-YA1R3 is shown as SEQ ID NO. 6; The nucleotide sequence of the CeNF-YA1F4 is shown as SEQ ID NO. 9; The nucleotide sequence of CeNF-YA1R4 is shown in SEQ ID NO.8; The nucleotide sequence of CeNF-YA1R5 is shown in SEQ ID NO.10.

[0009] This invention provides a method for amplifying the above-described technical solution. CeNF-YA1 Primer pairs for the gene, said primer pairs including CeNF-YA1F1 and CeNF-YA1R1; The nucleotide sequence of CeNF-YA1F1 is shown in SEQ ID NO.2; The nucleotide sequence of CeNF-YA1R1 is shown in SEQ ID NO.3.

[0010] The present invention provides the technical solution described above. CeNF-YA1 The application of the gene, the CeNF-YA1 protein, the expression vector, or the primer pair in the regulation of plant oils; The plants mentioned include Arabidopsis thaliana and / or tobacco.

[0011] Preferably, the regulation of plant oils includes: increasing the concentration of plant oils in the plant oil content of oils. CeNF-YA1 The level of gene expression promotes the accumulation of oil in plant seeds or plant tissues, thereby increasing the oil content.

[0012] Preferably, the plant tissue includes one or more of leaves, tubers, and rhizomes.

[0013] This invention provides a method for increasing the oil content of plants or promoting the accumulation of plant oils, comprising the following steps: The above technical solution is described CeNF-YA1 Genes are introduced into target plants to increase the oil content or promote oil accumulation in the target plants.

[0014] Preferably, the target plant includes: tobacco and / or Arabidopsis thaliana.

[0015] Beneficial effects: This invention provides for the first time CeNF-YA1 Genes, the ones mentioned CeNF-YA1 The nucleotide sequence of the gene is shown in SEQ ID NO.1, and its coding region is 792 bp in length.

[0016] This invention is the first to construct a system containing CeNF-YA1 A series of vectors, including subcellular localization of the gene, yeast hybridization, and plant overexpression, were developed, and for the first time it was demonstrated that the CeNF-YA1 protein has transcriptional activation function and is located in the cell nucleus, which conforms to the basic characteristics of transcription factors.

[0017] The present application proves that Figure 1 Overexpression of the gene can significantly increase the oil content of seeds and leaves of Arabidopsis thaliana; thus, it can be used to increase the oil content of seeds and vegetative tissues of plants and shows its application prospect in increasing the oil content of seeds and vegetative tissues. BRIEF DESCRIPTION OF DRAWINGS

[0018] In order to more clearly illustrate the technical solutions in the embodiments of the present application or the prior art, the drawings needed to be used in the embodiments will be briefly introduced below.

[0019] CeNF-YA1 For the present application CeNF-YA1 PCR amplification result of the gene: M: DNA marker III; CK: blank control; 1: Figure 2 ; CeNF-YA1 For the present application Figure 3 BLASTN alignment of the gene in NCBI GenBank database; CeNF-YA1 For the present application Figure 4 Sequence characteristics and evolutionary analysis of the encoded protein: A: analysis of the conserved domain of CeNF-YA1 protein; B: multiple sequence alignment of CeNF-YA1 and EgNF-YA3; C: evolutionary analysis of CeNF-YA1, EgNF-YA3 and NF-YA protein in Arabidopsis thaliana; Figure 5 For the present application Figure 6 For the present application Figure 7 For the present application P <0.01 significant difference; CeNF-YA1 For the present application CeNF-YA1Figure 1 shows the regulatory effect of gene overexpression on seed and leaf lipid accumulation in Arabidopsis thaliana: A: TAG content in transgenic and empty-transplant control T3 generation seeds; B: fold increase in TAG content in leaves of transgenic plants relative to empty-transplant T3 generation plants, showing the average of three biological replicates. "**" indicates based on... P The difference was significant (<0.01). Detailed Implementation The method for constructing the expression vector described in this invention has no special requirements and can be constructed using methods known in the art.

[0020] The NC kit described in this invention was purchased from Hainan Nixing Biotechnology Co., Ltd.; total RNA was extracted using the Tiangen plant polysaccharide and polyphenol RNA extraction kit (catalog number DP441); reverse transcription was performed using the Takara PrimeScript™ RTreagent Kit with gDNA Eraser kit (catalog number RR047A).

[0021] To further illustrate the present invention, the solutions provided by the present invention will be described in detail below with reference to the accompanying drawings and embodiments, but these should not be construed as limiting the scope of protection of the present invention.

[0022] In this invention, the primers used are shown in Table 1.

[0023] Table 1 Primer Information

[0024] The present invention CeNF-YA1The nucleotide sequence of the gene is shown in SEQ ID NO. 1, specifically: 5'-ATGGCCAAGCCTGTCTCCTCCTCAACTCAGTCCTCAAGTAGATCCCATCAAGATGATTCAGCTTCAGCTTTGAGTGATACTTCTGTCTATTCTGGTAGTTATGATAAGAGAAATGATCAGGTGAAATCAGCTTTATCCCTTGGCAATGGTGATTCCGCTTTTACTGCTCCAAAGCTTGATTATTTGCAGCCCTTTACATGCGTGCCGTACCCATATCCAGATGCCTGTTATGGCAGCGACATGACTGCATATGCTTCACCTCCGATGGTAAATGCACAGATGATGGGGATGGGGTCCAATTCGCGGGTGCCACTACCTCTCGAGCCTGCAGCAGAGGAACCAATATTTGTGAATGCAAAGCAGTACCATGCAATTCTTAGAAGGAGACAACTGCGTGCAAAGTTGGAGGCTGAAAATAAACTTGTGAAAGTCCGGAAGCCATATCTCCATGAATCACGGCATCTTCATGCGATGAAGCGGGCTCGGGGATCTGGTGGTCGGTTTCTTAATACCAAGCAGCAGTCTCAGCAGGGCCAGCTGTCAGACAGCCCAACACATGGCCTAAAAGTCTTAGGCACTAACATCGGACCTCAAGCTGAAGCCACTAATCAAACTATAGCAGGTGCTTCTGTGGCAGTAACTCCATCTAACTCCAGAAATGCACCAACCGAGCATAAGATCAACTATGGATCCCTGGATTTTAGAGGAGGGTTTCAAGCTGTAAATGGGTTTGGGAACAGAATTGGGATCAACAGTAGCTCCTCTCAGTGCGGTATCTCTATCATGCCGTGA-3'.

[0025] The amino acid sequence of the CeNF-YA1 protein according to the present application is shown in SEQ ID NO. 11, and specifically is: MAKPVSSSTQSSSRSHQDDSASALSDTSVYSGSYDKRNDQVKSALSLGNGDSAFTAPKLDYLQPFTCVPYPYPDACYGSDMTAYASPPMVNAQMMGMGSNSRVPLPLEPAAEEPIFVNAKQYHAILRRRQLRAKLEAENKLVKVRKPYLHESRHLHAMKRARGSGGRFLNTKQQSQQGQLSDSPTHGLKVLGTNIGPQAEATNQTIAGASVAVTPSNSRNAPTEHKINYGSLDFRGGFQAVNGFGNRIGINSSSSQCGISIMP*.

[0026] Example 1 Figure 1 Cloning and sequence analysis of the gene (1) Total RNA was extracted from tubers of Eleocharis dulcis cv. Reyan No. 3 at 10 different development stages, and then the mRNA was reversely transcribed into cDNA, which was used as a template for PCR amplification. The primers CeNF-YA1F1 (SEQ ID NO. 2) and CeNF-YA1R1 (SEQ ID NO. 3) were designed according to the full-length cDNA obtained from the transcriptome by using Primer Premier 5.0.

[0027] (2) After optimizing the PCR conditions according to the Tm value of the primers, the upstream primer CeNF-YA1F1 and the downstream primer CeNF-YA1R1 were used for PCR amplification with the above-mentioned reversely transcribed cDNA as a template. The PCR amplification reaction system is shown in Table 2. The PCR reaction program was as follows: 98℃ for 3 min; 98℃ for 30 sec, 60℃ for 30 sec, 72℃ for 50 sec (35 cycles); 72℃ for 5 min; and 4℃ storage.

[0028] Table 2 PCR amplification reaction system

[0029] (3) One specific band of about 800 bp was obtained by PCR amplification CeNF-YA1 ), which was recovered by using an OMEGA gel recovery kit (item number D2500-02) and then cloned into the cloning vector pMD19-T (item number 6013) of Takara. After blue-white spot screening and colony PCR verification, sequence determination was performed.

[0030] (4) Sequence analysis showed that Figure 2The coding region (CDS) of CeNF-YA1 is 792 bp in length, with a GC content of 47.47%, and is predicted to encode 263 amino acids, with a theoretical molecular weight of 28.31 kDa, an isoelectric point (pI) of 9.51, a total average hydrophobicity index (GRAVY) of -0.573, an instability index (II) of 57.22, and an aliphatic index (AI) of 63.88.

[0031] (5) BLASTN was used to align the NCBI GenBank database, and no similar sequences were found Figure 3 ), confirming that the cloned gene is a new gene.

[0032] (6) CDD (https: / / www.ncbi.nlm.nih.gov / Structure / cdd / wrpsb.cgi?) analysis and sequence alignment showed that CeNF-YA1 protein contains a conserved CBFB_NFYA domain Figure 3 A and 3B). To further reveal the evolutionary characteristics of CeNF-YA1, MEGA6 (https: / / www.megasoftware.net / ) was used to construct a phylogenetic tree with EgNF-YA3 and NF-YA proteins in Arabidopsis thaliana, and the results are shown in Figure 3 C.

[0033] It can be seen from Figure 4 C that CeNF-YA1 is clustered with EgNF-YA3 (Egu032172), AtNF-YA5 (AT1G54160), AtNF-YA6 (AT3G14020), AtNF-YA3 (AT1G72830), and AtNF-YA8 (AT1G17590), with sequence similarities of 50.54%, 44.70%, 39.46%, 42.70%, and 38.07%, respectively.

[0034] Example 2 Subcellular localization analysis of CeNF-YA1 protein (1) WoLF PSORT (https: / / www.genscript.com / wolf-psort.html) analysis showed that CeNF-YA1 may be localized in the nucleus.

[0035] (2) In order to confirm the above prediction, the primer pair CeNF-YA1F2 (SEQ ID NO. 4) and CeNF-YA1R2 (SEQ ID NO. 5) shown in Table 1 was designed near the start and stop codons of the sequenced gene, and PCR amplification was performed with the PCR product diluted 100 times in Example 1 as a template (the system and procedure of PCR amplification were the same as those in Example 1). After PCR amplification, the target fragment was recovered by gel cutting and cloned into pNC-Cam1304-SubN using an NC kit (Hainan Youxing Biotechnology Co., Ltd.) to construct a fusion expression vector pNC-Cam1304-CeNF-YA1 with EGFP, wherein the NC ligation system is shown in Table 3.

[0036] Table 3 NC ligation system

[0037] (4) The recombinant plasmid pNC-Cam1304-CeNF-YA1 was transformed into Agrobacterium tumefaciens strain GV3101 (containing pSoup-P19), and the specific process was as follows: (4-1) 1 μg of nuclear localization positive plasmid pNC-Cam1304-H2A and pNC-Cam1304-CeNF-YA1 were added to 100 μL competent cells GV3101, respectively, and mixed uniformly by blowing with a pipette gun; (4-2) ice bath for 5 min, frozen in liquid nitrogen for 5 min, then 37°C water bath for 5 min, and ice for 5 min; (4-3) add 700 μL of LB medium without antibiotics, 28°C, 200 rpm, and shake culture for 2 h; (4-4) the bacteria were inoculated on solid LB medium containing 50 mg / L kanamycin and 100 mg / L rifampicin, and incubated at 28°C for 2-3 days; (4-5) single colonies were picked for colony PCR detection, and positive clones were selected for genetic transformation experiments.

[0038] (5) 4-week-old N. benthamiana leaves were transformed by microinjection, and the specific process was as follows: (5-1) the above positive bacteria were inoculated in 2 mL of LB liquid medium (50 mg / L rifampicin) and incubated at 28°C, 210 rpm overnight; (5-2) 1 mL of bacterial solution was taken in 30 mL of LB liquid medium and incubated at 28°C, 210 rpm until the bacterial solution concentration OD600 was 0.8; (5-3) 5000 rpm centrifugal 3 min to collect bacterial body, after removing the supernatant, add resuspension liquid (10 mmol / L MgCl2, 0.2 mmol / L acetyl syringone (AS) and 10 mmol / L MES, use KOH to adjust the PH to 5.6), resuspend the bacterial body, repeat twice, and adjust the concentration of the resuspended bacterial solution to OD600 0.6; (5-4) The resuspended bacterial solution is incubated at 28°C for 3-5 h, and the lower epidermis of tobacco is injected (one gene injection is one piece of tobacco leaf), and the lower epidermis of the leaf can be slightly scratched with a needle before injection; (5-5) After the injection is completed, the injected tobacco leaves are incubated in the dark for 2-4 d, and the injected tobacco leaves are sectioned for fluorescence signal detection; (6) Laser confocal microscope observation shows that the fluorescence signals of the experimental group (green fluorescence) and the positive control (red fluorescence) are found in the nucleus, and the fluorescence signals are highly coincident, indicating that CeNF-YA1 plays a role in the nucleus Figure 5 ).

[0039] Example 3 Identification of the transcriptional activation function of CeNF-YA1 protein (1) To identify the transcriptional activation function of CeNF-YA1 and the position of the activation domain, primers as shown in Table 1 are designed near the CBFB_NFYA domain; the PCR product in Example 1 is diluted 100 times as a template, and CeNF-YA1F2 (SEQ ID NO. 4) and CeNF-YA1R2 (SEQ ID NO. 5) are used as primers to amplify the full-length coding region of CeNF-YA1 (SEQ ID NO. 1), after PCR amplification, the gel is recovered and cloned into pNC-GBKT7 to construct the yeast two-hybrid bait vector pNC-GBKT7-CeNF-YA1F; The PCR product in Example 1 is diluted 100 times as a template, and CeNF-YA1F2 (SEQ ID NO. 4) and CeNF-YA1R3 (SEQ ID NO. 6) are used as primers to amplify the N1 end of CeNF-YA1 (i.e. nucleotide sequence 1-327 bp shown in SEQ ID NO. 1), after PCR amplification, the gel is recovered and cloned into pNC-GBKT7 to construct the yeast two-hybrid bait vector pNC-GBKT7-CeNF-YA1N1; The PCR product in Example 1 was diluted 100 times as a template, and CeNF-YA1F2 (SEQ ID NO. 4) and CeNF-YA1R4 (SEQ ID NO. 8) were used as primers to amplify the N2 end of CeNF-YA1 (i.e. 1-510 bp of the nucleotide sequence shown in SEQ ID NO. 1, containing the CBFB_NFYA domain). After PCR amplification, the gel was cut and recovered, and then cloned into pNC-GBKT7 to construct the yeast two-hybrid bait vector pNC-GBKT7-CeNF-YA1N2. The PCR product in Example 1 was diluted 100 times as a template, and CeNF-YA1F3 (SEQ ID NO. 7) and CeNF-YA1R2 (SEQ ID NO. 5) were used as primers to amplify the C end of CeNF-YA1 (i.e. 511-792 bp of the nucleotide sequence shown in SEQ ID NO. 1, downstream of the CBFB_NFYA domain). After PCR amplification, the gel was cut and recovered, and then cloned into pNC-GBKT7 to construct the yeast two-hybrid bait vector pNC-GBKT7-CeNF-YA1C. The PCR amplification method and NC ligation method are the same as those in Example 2.

[0040] (2) The empty vector (pNC-GBKT7) and the four recombinant plasmids (pNC-GBKT7-CeNF-YA1F, pNC-GBKT7-CeNF-YA1N1, pNC-GBKT7-CeNF-YA1N2 and pNC-GBKT7-CeNF-YA1C) were respectively transformed into Y2H-Gold yeast cells, and then the transformed yeast cells were selected on a tryptophan-deficient medium. Positive bacteria were selected and screened on a tryptophan- and histidine-deficient medium, and then the positive bacteria were subjected to X-α-gal color development reaction.

[0041] (3) The results showed that the engineered bacteria transformed with pNC-GBKT7-CeNF-YA1F and pNC-GBKT7-CeNF-YA1N1, pNC-GBKT7-CeNF-YA1N2 could survive on the selective medium and develop blue color, while the yeast cells transformed with the empty vector and pNC-GBKT7-CeNF-YA1C failed to exhibit β-galactosidase activity. This indicated that CeNF-YA1 has a transcriptional activation function, and the activation domain is located upstream of the CBFB_NFYA domain. CeNF-YA1 ).

[0042] Example 4 CeNF-YA1 Identification of lipid regulation function (1) To identify the function of CeNF-YA1 in lipid regulation, the yeast two-hybrid system was used to screen the protein interaction of CeNF-YA1 and the lipid regulation-related proteins. Figure 6To investigate its function in regulating plant oils, the above PCR product was diluted 100-fold as a template and CeNF-YA1F4 (SEQ ID NO. 9) and CeNF-YA1R5 (SEQ ID NO. 10) were used as primers for PCR amplification. The target fragment was then recovered from the gel and cloned into the plant overexpression vector pCAMBIA1301 using homologous recombination to construct the recombinant vector pCAMBIA1301-CeNF-YA1. (2) The recombinant vector pCAMBIA1301-CeNF-YA1 and the empty pCAMBIA1301 were separately transformed into GV3101 (containing pSoup-P19), and 4-week-old tobacco leaves were transformed by micro-injection method; samples were collected 1 day, 3 days and 5 days after transformation for the determination of oil content. (3) Oil content determination results based on Agilent 7890A (HP-FFAP, 30 9 0.25 mm ID, 0.25 lm, Santa Clara, CA, USA) showed that, compared with the control group converted to empty loading, the TAG content in the experimental group increased by 1.52, 1.57 and 2.05 times after 1 day, 3 days and 5 days of conversion, respectively. CeNF-YA1 ).

[0043] Example 5 CeNF-YA1 Application in the regulation of lipids in seeds and vegetative tissues (1) For evaluation Figure 7 To explore its potential application in regulating plant oils, the engineered bacteria containing the recombinant vector pCAMBIA1301-CeNF-YA1 or the empty vector pCAMBIA1301 were used to transform wild-type Arabidopsis thaliana using the inflorescence immersion method. The transformed plants were then cultured under the same conditions until maturity and seed harvesting. T1 seeds were screened for resistance on MS solid plates containing 25 µg / mL hygromycin to obtain T2 generation positive transgenic plants. After maturation, these plants underwent further resistance screening to obtain stably inherited T3 generation positive transgenic plants.

[0044] (2) To analyze the effect of gene overexpression on seed oil regulation, oil content was measured in T3 generation transgenic Arabidopsis seeds. The results showed that the oil content of the transgenic seeds was 32.51%, which was significantly higher than that of the empty-vectored seeds (27.50%). CeNF-YA1 A); (3) For evaluation CeNF-YA1 The application value of this study in regulating oil content in plant vegetative tissues was investigated. Leaves from the aforementioned T3 generation transgenic Arabidopsis thaliana were collected for oil content determination. The results showed that, compared to the control group (transformed into an empty vector), oil content was significantly higher. Figure 7 The oil content of the leaves increased by 3.57 times due to overexpression. CeNF-YA1 B).

[0045] (4) The above results show that, ​ The application has the function of promoting oil accumulation and shows its application prospect in improving the oil content of seeds and nutrient tissues.

[0046] Although the above embodiments have made a detailed description of the present application, it is only a part of the embodiments of the present application, not all the embodiments, and other embodiments can be obtained according to the present embodiments without creativity, which all belong to the protection scope of the present application.

Claims

1. CeNF-YA1 A gene characterized in that, The CeNF-YA1 The nucleotide sequence of the gene is shown as SEQ ID NO.

1.

2. The method of claim 1 CeNF-YA1 CeNF-YA1 protein encoded by the gene, characterized in that, The amino acid sequence of the CeNF-YA1 protein is shown as SEQ ID NO.

11.

3. A composition comprising the polypeptide of claim 1 CeNF-YA1 An expression vector comprising the gene of claim 1, characterized in that, The expression vector comprises one or more of the following: cell localization vector pNC-Cam1304-CeNF-YA1, yeast hybridization vector and plant overexpression vector pCAMBIA1301-CeNF-YA1. The yeast hybridization vector is one or more of the following: pNC-GBKT7-CeNF-YA1F, pNC-GBKT7-CeNF-YA1N1, pNC-GBKT7-CeNF-YA1N2 and pNC-GBKT7-CeNF-YA1C.

4. The expression vector of claim 3, wherein, The pNC-Cam1304-CeNF-YA1 is constructed with CeNF-YA1F2 and CeNF-YA1R2; The pNC-GBKT7-CeNF-YA1F is constructed with CeNF-YA1F2 and CeNF-YA1R2; The pNC-GBKT7-CeNF-YA1N1 is constructed with CeNF-YA1F2 and CeNF-YA1R3; The pNC-GBKT7-CeNF-YA1N2 is constructed with CeNF-YA1F2 and CeNF-YA1R4; The pNC-GBKT7-CeNF-YA1C is constructed with CeNF-YA1F3 and CeNF-YA1R2; The pCAMBIA1301-CeNF-YA1 is constructed with CeNF-YA1F4 and CeNF-YA1R5; The nucleotide sequence of the CeNF-YA1F2 is shown as SEQ ID NO. 4; The nucleotide sequence of the CeNF-YA1R2 is shown as SEQ ID NO. 5; The nucleotide sequence of the CeNF-YA1F3 is shown as SEQ ID NO. 7; The nucleotide sequence of the CeNF-YA1R3 is shown as SEQ ID NO. 6; The nucleotide sequence of the CeNF-YA1F4 is shown as SEQ ID NO. 9; The nucleotide sequence of the CeNF-YA1R4 is shown as SEQ ID NO. 8; The nucleotide sequence of the CeNF-YA1R5 is shown as SEQ ID NO.

10.

5. A primer pair for amplifying the gene of claim 1, characterized in that, CeNF-YA1 The primer pair comprises CeNF-YA1F1 and CeNF-YA1R1; ​ The nucleotide sequence of the CeNF-YA1F1 is shown as SEQ ID NO. 2; The nucleotide sequence of the CeNF-YA1R1 is shown as SEQ ID NO.

3.

6. The use of the gene of claim 1, the CeNF-YA1 protein of claim 2, the expression vector of claim 3 or 4, or the primer pair of claim 5 in the regulation of plant oil. CeNF-YA1 The gene of claim 1, the CeNF-YA1 protein of claim 2, the expression vector of claim 3 or 4, or the primer pair of claim 5 in the regulation of plant oil. The plants include: Arabidopsis and / or tobacco.

7. Use according to claim 6, characterized in that, The regulation of plant oils includes: increasing the concentration of oils in plants. CeNF-YA1 The level of gene expression promotes the accumulation of oil in plant seeds or plant tissues, thereby increasing the oil content.

8. Use according to claim 7, characterized in that, The plant tissue comprises one or more of the following: leaf, tuberous root and tuber.

9. A method for increasing lipid content or promoting lipid accumulation in a plant, comprising, The method comprises the following steps: The method of claim 1 CeNF-YA1 introducing the gene into a target plant to increase oil content or promote oil accumulation in the target plant.

10. The method of claim 9, wherein, The target plant comprises tobacco and / or Arabidopsis.

Images