Strawberry FaLEC2 gene and application thereof

Abstract

The invention discloses a strawberry FaLEC2 gene and application thereof, and relates to the field of plant molecular biology, the nucleotide sequence of the FaLEC2 gene is as shown in SEQ ID NO.1, and the amino acid sequence of protein encoded by the strawberry FaLEC2 gene is as shown in SEQ ID NO.2. The invention studies the application of the FaLEC2 gene in regulating and controlling the contents of vinblastine, 3-deoxy-D-mann-2-octanone acid, raspberry glycoside and linolenic acid in strawberries. It is found that through overexpression of the FaLEC2 gene, the content of vinblastine, the content of 3-deoxy-D-mann-2-octanone acid, the content of raspberry glycoside and the content of linolenic acid in the strawberries can be increased; the metabolism regulation and control mechanism of the vinblastine, the 3-deoxy-D-mann-2-octanone acid, the raspberry glycoside and the linolenic acid in the strawberry fruits can be further known, so that the content of the vinblastine, the 3-deoxy-D-mann-2-octanone acid, the raspberry glycoside and the linolenic acid in the strawberry fruits can be regulated and controlled by utilizing a biotechnological means; an important foundation is laid for strawberry variety improvement.

Description

Technical Field

[0001] The invention relates to the field of plant molecular biology, in particular to a strawberry FaLEC2 gene and application thereof. Background Art

[0002] Strawberry fruit is rich in nutrients and has a unique aroma. It is known as the "Queen of Fruits". It is rich in bioactive substances such as anthocyanins and flavonoids and is gradually regarded as a functional fruit that is beneficial to the human body. It is widely planted around the world and has important economic value. my country has become the largest producer and consumer of strawberries. Further improving the quality of strawberry fruit will help meet the growing material needs of consumers.

[0003] Strawberry fruit contains certain ingredients that are beneficial to human health, such as vinblastine, 3-deoxy-D-manno-2-octulose acid, raspberry glycoside, and α-linolenic acid. Vinblastine is a derivative product of the biosynthesis pathway of terpenoid indole alkaloids, and has anti-inflammatory, anti-tumor, antibacterial, and anti-allergic functions; 3-deoxy-D-manno-2-octulose acid is a scarce monosaccharide and an important component of cell wall lipopolysaccharide. It connects lipids and core oligosaccharides and plays an important role in maintaining the integrity of cell walls and outer membranes; raspberry glycoside is a natural ketone compound with a strong fruity aroma and certain antioxidant and anti-inflammatory activities; α-linolenic acid is one of the essential fatty acids for people. The human body cannot synthesize it by itself and must be taken in through food to meet its needs. It is of great significance to human health. Increasing the content of vinblastine, 3-deoxy-D-manno-2-octulose acid, raspberry glycoside, and linolenic acid in strawberries will help further improve the nutritional value and economic value of strawberries. Summary of the invention

[0004] The present invention aims to provide a strawberry FaLEC2 gene and an application thereof, so as to increase the content of vinblastine, 3-deoxy-D-manno-2-octulose acid, raspberry glycoside and linolenic acid in strawberries.

[0005] In order to achieve the above object, the specific scheme adopted by the present invention is: a strawberry FaLEC2 gene, the nucleotide sequence of the FaLEC2 gene is shown in SEQ ID NO.1.

[0006] SEQ ID NO.1:

[0007] atggagaacttccatcaacctttctttccttccacaacgaccaccatgacccccatttatgaacctatgagttcctc cccttcaattcttgctagttcctctatgtccaattattatccgagagttcagtacctaccaccaaccaatggcagcggccagttgtattttcaaagcttgtaccctcatttccagaagcaggcctctcagccacccttccaatatcgcaggcatcccctcctcctccctcggccacagccaaatccagtctctggagatcaaggagggattgagtccaaggctgcaagagatgagcgaaggatggcacgccaaagaaggctcttttcaagcagaagcgctgctgactctgtttcctcatcaagtccaacgcagatggatttatgtggtgctgattctgataggcagaacacagacctctgcacccgcaccccggatatgaaggaactgaaaaagttgttcaagaaggagttgaagaagagtgatgttggttgctggggcaggattgtcattccaaagaaggaggcagagaaataccttccaagccttgttgaaaaacaaggcatcatgctgtggatcagagatgtccattccagtcatgtatggggcatgaaatacaagttgttgtcactgtactgggcaaataacaaaagtcgaatgtatgtcttcgaaagtgcaggtaagcctgagaggacctga

[0008] A protein encoded by the strawberry FaLEC2 gene, the amino acid sequence of the protein is shown in SEQ ID NO.2.

[0009] SEQ ID NO.2:

[0010] MENFHQPFFPSTTTTMTPIYEPMSSSPSILASSSMSNYYPRVQYLPPPTN GSGQLYFQSLYPHFQKQASQPPFQYRRHPLLLPRPQPNPVSGDQGGIESKAARDERRMARQRRLFSSRSAADSVSSSSPTQMDLCGADSDRQNTDLCTRTPDMKELKKLFKKELKKSDVGCWGRIVIPKKEAEKYLPSLVEKQGIMLWIRDVHSSHVWGMKYKLLSLYWKPPRMYVFESAGERT

[0011] The invention discloses an application of a strawberry FaLEC2 gene in regulating the content of vinblastine, 3-deoxy-D-manno-2-octulose acid, raspberry glycoside or linolenic acid in strawberry fruit.

[0012] As a further optimization of the above technical solution: first construct an overexpression vector of the strawberry FaLEC2 gene; then transform the overexpression vector into Agrobacterium; finally, infect the strawberry fruit with the transformed Agrobacterium.

[0013] As a further optimization of the above technical solution: first construct an overexpression vector of the strawberry FaLEC2 gene; then transform the overexpression vector into Agrobacterium; finally, immerse the strawberry tissue culture seedlings in the Agrobacterium liquid carrying the FaLEC2 gene for infection, and subculture to obtain transgenic strawberry plants.

[0014] As a further optimization of the above technical solution: the overexpression vector is a plasmid containing pC1300s-GFP-FaLEC2.

[0015] As a further optimization of the above technical solution: the Agrobacterium is GV3101 Agrobacterium.

[0016] As a further optimization of the above technical solution: the overexpression vector is transformed into Agrobacterium using the freeze-thaw method.

[0017] Compared with the prior art, the present invention has the following beneficial effects:

[0018] The invention provides a FaLEC2 gene cloned from strawberry, and studies the application of the FaLEC2 gene in regulating the contents of vinblastine, 3-deoxy-D-manno-2-octulose acid, raspberry glycoside and linolenic acid in strawberry. It is found that transient overexpression of the FaLEC2 gene can increase the contents of vinblastine, 3-deoxy-D-manno-2-octulose acid, raspberry glycoside and linolenic acid in strawberry, which is helpful to further understand the metabolic regulation mechanism of vinblastine, 3-deoxy-D-manno-2-octulose acid, raspberry glycoside and linolenic acid in strawberry fruit, so that the contents of vinblastine, 3-deoxy-D-manno-2-octulose acid, raspberry glycoside and linolenic acid in strawberry fruit can be regulated by biotechnology means, thereby laying an important foundation for strawberry variety improvement. BRIEF DESCRIPTION OF THE DRAWINGS

[0019] Figure 1 These are the electrophoresis images and double enzyme digestion images of the strawberry FaLEC2 gene overexpression vector: 1 is the FaLEC2 gene overexpression vector; 2 is the double enzyme digestion of the FaLEC2 gene overexpression vector; M is Maker;

[0020] Figure 2 is the relative expression level of FaLEC2 gene in FaLEC2 gene overexpressing strawberry fruits and control fruits;

[0021] Figure 3 These are the metabolites that change significantly in strawberry fruit with overexpression of the FaLEC2 gene. DETAILED DESCRIPTION

[0022] The technical solution of the present invention is further elaborated in detail below in conjunction with specific embodiments. The parts of the present invention that are not described and disclosed in detail in the following embodiments should be understood as the prior art known or should be known to those skilled in the art.

[0023] Example 1 Cloning of strawberry FaLEC2 gene

[0024] 1) Collect the small green fruits of 'Zhangji' strawberry, cut them and immediately place them in liquid nitrogen and store them in a -80℃ refrigerator for later use.

[0025] 2) RNA was extracted from strawberry fruit during the small green period according to the instructions of RNAprep Pure Polysaccharide and Polyphenol Plant Total RNA Extraction Kit (Tiangen Biochemical Technology Co., Ltd., Beijing, China). The RNA quality and concentration were tested using Nanodrop 2000 Nucleic Acid Analyzer. The RNA OD 260 / OD 280 The value is between 1.8 and 2.0. RNA integrity was observed using 1% agarose gel electrophoresis, and the brightness of the 28S rRNA band was required to be close to twice that of the 18S rRNA band. TMRT reagent Kit with gDNA Eraser Kit (Takara, Japan) instructions for reverse transcription to synthesize the first strand of cDNA.

[0026] 3) Using the above cDNA as a template, primers FaLEC2-F (5'-agctttcgcgagctcggtaccATGGAGAACTTCCATCAACCTTTC-3') and FaLEC2-R (5'-gcccttgctcaccatggatccTCAGGTCCTCTCAGGCTTACCTG-3') were used to generate the PCR product using a high-fidelity polymerase system. Max Master Mix (Nanjing Novogene Biotech Co., Ltd., Nanjing, China) was used for PCR reaction to obtain the FaLEC2 gene amplification product. The PCR reaction system was: 12.5 μL 2×Phanta Max Master Mix, 1 μL FaLEC2-F (10 μM), 1 μL FaLEC2-R (10 μM), 1 μL cDNA, 9.5 μL ddH 2 O; PCR reaction program: 95℃ pre-denaturation for 3min; 95℃ denaturation for 15s, 60℃ annealing for 15s, 72℃ extension for 1.5min, 35 cycles; 72℃ extension for another 5min. The amplified products were detected by 1% agarose gel electrophoresis, and the target fragment of FaLEC2 gene was recovered and purified according to the instructions of DNA gel recovery kit (Takara, Japan).

[0027] 4) The pC1300s-GFP vector was double-digested with KpnI and BamHI at 37°C for 4 h, and the pC1300s-GFP vector digested fragment was recovered and purified according to the instructions of the DNA gel recovery kit (Takara, Japan). IIOne Step Cloning Kit (Nanjing Novogene Biotech Co., Ltd., Nanjing, China) Instructions: The purified FaLEC2 gene fragment was ligated to the pC1300s-GFP vector to obtain the pC1300s-GFPFaLEC2 plasmid. The ligation product was transformed into Escherichia coli DH5α competent cells by heat shock method, and coated with solid LB medium containing 50 μg / mL kanamycin. The cells were cultured at 37°C until a single colony with a diameter of 1-2 mm was grown. A single colony was picked and suspended in 10 ul of sterile water. The colony suspension was used as a template and 2×T5Super PCR was performed. Mix (Basic) (Beijing Qingke Biotechnology Co., Ltd., Beijing, China) instruction manual, colony PCR was performed using the FaLEC2 gene upstream primer FaLEC2-F (5'-agctttcgcgagctcggtaccATGGAGAACTTCCATCAACCTTTC-3') and the pC1300s-GFP vector reverse primer pC1300s-GFP-R (5'-CCGTCCAGCTCGACCAGGAT-3'), and the PCR product was subjected to 1% agarose gel electrophoresis. The single colony suspension corresponding to the band position of 714 bp was cultured at 37°C for 12 h using LB liquid medium containing 50 μg / mL kanamycin, and the bacterial liquid was sent to a biological company for sequencing. The sequencing results confirmed that the obtained was the full-length strawberry FaLEC2 cDNA (SEQ ID NO: 1). The pC1300s-GFP-FaLEC2 plasmid was digested with SacI and EcoRI for identification, as shown in Figure 2. Figure 1 shown.

[0028] Example 2 Application of the strawberry FaLEC2 gene in regulating the content of vinblastine, 3-deoxy-D-manno-2-octulose acid, raspberry glycoside or linolenic acid in strawberry fruit

[0029] 1) Construction of strawberry FaLEC2 gene overexpression vector

[0030] The strawberry FaLEC2 gene cloned in Example 1, whose encoded protein amino acid sequence is shown in SEQ ID NO. 2, was used to construct a plasmid containing pC1300s-GFP-FaLEC2 as an overexpression vector of the strawberry FaLEC2 gene.

[0031] II) Agrobacterium transformation containing FaLEC2 gene overexpression vector

[0032] 5 μL of plasmid containing pC1300s-GFP-FaLEC2 was used to transform 100 μL of Agrobacterium tumefaciens GV3101 by freeze-thaw method, gently pipetted to mix, placed in ice bath for 30 min, frozen in liquid nitrogen for 5 min, heat-shocked in a 37°C water bath for 5 min, and then added to 1 mL of LB liquid medium and cultured in a 28°C incubator for 2 h. Then 100 μL of bacterial solution was spread on LB solid medium containing 50 μg / mL kanamycin and 25 μg / mL rifampicin, inverted and cultured in a 28°C incubator until a single colony with a diameter of 1-2 mm was grown. Colony PCR was performed according to the instructions of 2×T5Super PCR Mix (Basic) (Beijing Qingke Biotechnology Co., Ltd., Beijing, China) to identify and obtain GV3101 Agrobacterium containing pC1300s-GFP-FaLEC2.

[0033] III) Agrobacterium-mediated transformation of strawberry fruit

[0034] Select the small green period fruits of 'Zhangji' strawberry and use the injection method to infect. The specific method is as follows:

[0035] 1) Pick the positive clone containing pC1300s-GFP-FaLEC2 plasmid, inoculate it into 4 mL LB liquid medium (50 μg / mL kanamycin and 25 μg / mL rifampicin), shake the culture at 28°C and 200 rpm for 12 h to obtain bacterial solution I;

[0036] 2) Inoculate the above bacterial solution I into 100 mL LB liquid medium containing kanamycin (100 μL) and rifampicin (50 μL) at a ratio of 1:100, and culture at 28°C with shaking at 200 rpm until OD 600 =1.0, transfer to a 50 mL centrifuge tube, centrifuge at 4000 rpm for 10 min, and discard the supernatant to obtain bacterial solution II;

[0037] 3) Add 20 mL of 4°C precooled sterilized ultrapure water to bacterial solution II, mix well, centrifuge at 4°C, 4000 rpm for 10 min, discard the supernatant, and obtain bacterial solution III;

[0038] 4) Add 20 mL of 4°C precooled sterilized ultrapure water to bacterial solution III, mix well, centrifuge at 4°C, 4000 rpm for 10 min, discard the supernatant, and obtain bacterial solution IV;

[0039] 5) Preparation of infection solution: weigh MgCl 2 6H 2 O 2.033g, fatty acid methyl ester sulfonate 2.172g, dissolved in 1L H 2 O, adjust pH to 5.7, sterilize at 121℃ for 15min, this is the infection solution, and it is ready for use;

[0040] 6) Add acetosyringone to the infection solution to 150 μmol / L, pre-cool, and use the pre-cooled infection solution to suspend the bacterial solution IV to OD 600 =1.0~1.5, place at 25℃ for 3h to obtain bacterial suspension containing pC1300s-GFP-FaLEC2, use a 1mL syringe to draw the bacterial suspension and insert it into the strawberry fruit from the strawberry stalk, and push the syringe very slowly until the whole strawberry fruit is soaked;

[0041] 7) Leave the injected strawberry fruits on the plants to grow naturally for 7 days before picking them.

[0042] Example 3 Application of the strawberry FaLEC2 gene in regulating the content of vinblastine, 3-deoxy-D-manno-2-octulose acid, raspberry glycoside or linolenic acid in strawberry fruit

[0043] The overall steps of constructing the strawberry FaLEC2 gene transient expression vector and Agrobacterium transformation in this embodiment are the same as those in Example 2, except that: in this embodiment, the strawberry tissue culture seedlings are first immersed in the Agrobacterium liquid carrying the FaLEC2 gene for infection; then the tissue culture seedlings are transferred to the regeneration medium for culture; then the tissue culture seedlings are transferred to the differentiation medium for culture to obtain resistant buds; finally, the resistant buds are rooted on the rooting medium, and then the seedlings are hardened and transplanted to obtain transgenic strawberry plants. The specific methods of immersion and obtaining transgenic strawberry plants are all technical, and will not be repeated here.

[0044] Comparative Example 1

[0045] The steps of this comparative example are generally the same as those of Example 2, except that in this comparative example, a blank impregnation liquid (i.e., an impregnation liquid without a FaLEC2 gene overexpression vector) is used to impregnate strawberries, as follows:

[0046] 5 μL of plasmid containing pC1300s-GFP was used to transform 100 μL of Agrobacterium tumefaciens GV3101 by freeze-thaw method, gently pipetted to mix, ice bathed for 30 min, frozen in liquid nitrogen for 5 min, heat-shocked in a 37°C water bath for 5 min, and then added to 1 mL of LB liquid medium and cultured in a 28°C incubator for 2 h. Then 100 μL of bacterial solution was spread on LB solid medium containing 50 μg / mL kanamycin and 25 μg / mL rifampicin, and inverted in a 28°C incubator to grow a single colony with a diameter of 1-2 mm. Colony PCR was performed according to the instructions of 2×T5Super PCR Mix (Basic) (Beijing Qingke Biotechnology Co., Ltd., Beijing, China) to identify and obtain GV3101 Agrobacterium containing pC1300s-GFP.

[0047] Select the small green period fruits of 'Zhangji' strawberry and use the injection method to infect. The specific method is as follows:

[0048] 1) Pick positive clones containing the pC1300s-GFP plasmid and inoculate them into 4 mL of LB liquid medium (50 μg / mL kanamycin and 25 μg / mL rifampicin). Shake the culture at 28 °C and 200 rpm for 12 h to obtain bacterial liquid I.

[0049] 2) Inoculate the above bacterial liquid I into 100 mL of LB liquid medium containing kanamycin (100 μL) and rifampicin (50 μL) at a ratio of 1:100. Incubate the culture at 28 °C and 200 rpm until OD 600 = 1.0, then transfer the bacterial liquid to a 50 mL centrifuge tube, centrifuge at 4000 rpm for 10 min, and discard the supernatant to obtain bacterial liquid II.

[0050] 3) Add 20 mL of pre-cooled sterile ultrapure water at 4 °C to bacterial liquid II, mix well, centrifuge at 4 °C and 4000 rpm for 10 min, and discard the supernatant to obtain bacterial liquid III.

[0051] 4) Add 20 mL of pre-cooled sterile ultrapure water at 4 °C to bacterial liquid III, mix well, centrifuge at 4 °C and 4000 rpm for 10 min, and discard the supernatant to obtain bacterial liquid IV.

[0052] 5) Preparation of infection solution: Weigh 2.033 g of MgCl 2 ·6H 2 O and 2.172 g of fatty acid methyl ester sulfonate, dissolve them in 1 L of H 2 O, adjust the pH to 5.7, and sterilize at 121 °C for 15 min to obtain the infection solution, which is reserved for use.

[0053] 6) Add acetosyringone to the infection solution to a concentration of 150 μmol / L, pre-cool it, suspend bacterial liquid IV with the pre-cooled infection solution until OD 600 = 1.0 - 1.5, place it at 25 °C for 3 h to obtain a bacterial liquid suspension containing pC1300s-GFP, and insert a 1 mL syringe into the strawberry fruit from the fruit stalk, and very slowly push the syringe until the entire strawberry fruit is soaked.

[0054] 7) Keep the injected strawberry fruits on the plants and let them grow naturally for 7 days, then pick them.

[0055] Analysis of test results

[0056] For easy understanding, set Example 2 as the test group and Comparative Example 1 as the control group, and detect the strawberry fruits of the test group and the control group.

[0057] <Verification of FaLEC2 gene overexpressing strawberry fruits>

[0058] According to PrimeScript TMRT-PCR Kit (Takara, Japan) instruction manual. Detect the expression levels of the FaLEC2 gene in the infected strawberry fruits of the control group and the experimental group. The primers used are FaLEC2-F (5’-GCTGTGGATCAGAGATGTCC-3’) / FaLEC2-R (5’-GCTTACCTGCACTTTCGAAG-3’), FaActin-F (5’-GACAGGTGTTGTGGTAGACG-3’) / FaActin-R (5’-ACGGACTTGATGCTGCTCCC-3’). The results are as follows Figure 2 , the expression level of the FaLEC2 gene in the fruits of the experimental group was significantly increased compared with that of the control group, indicating that the FaLEC2 gene was overexpressed in strawberry fruits.

[0059] <Determination of metabolites in strawberry fruits overexpressing the FaLEC2 gene>

[0060] The strawberry fruits with FaLEC2 gene overexpression in the experimental group and the fruits in the control group were ground into powder, 50 mg of powder was weighed, and 1200 μL of 70% methanol / water (1:1) internal standard extract precooled at -20°C was added. The internal standard extract was prepared by dissolving 1 mg of standard in 1 mL of 70% methanol water to prepare 1000 μg / mL standard stock solution, and the 1000 μg / mL stock solution was further diluted with 70% methanol to prepare 250 μg / mL internal standard solution; vortex once every 30 minutes, each time for 30 seconds, and vortexed 6 times in total; centrifuged at 12000 rpm for 3 minutes, the supernatant was aspirated, and the sample was filtered with a 0.22 μm pore size microporous filter membrane and stored in an injection bottle. The separation was performed using an ultra-high performance liquid chromatography system equipped with an ACQUITY Premier HSS T3 Column (1.8 μm, 2.1 mm×100 mm, Waters, UK) chromatographic column. Mobile phase A was 0.1% formic acid / water; mobile phase B was 0.1% formic acid / acetonitrile; column temperature was 40°C, flow rate was 0.4 mL / min, injection volume was 4 μL; gradient elution program: 0-2 min, 80% mobile phase A; 2-5 min, 5-60% mobile phase B; 6-7.5 min, 99% mobile phase B; 7.6-10 min, 1-95% mobile phase A. The eluent was collected in positive and negative ion modes using a tandem mass spectrometer AB TripleTOF (Waters, UK); in positive ion mode, the ionization voltage was 5000 V and the ion source temperature was 550°C; in negative ion mode, the ionization voltage was -4000 V and the ion source temperature was 450°C; data were collected in ESI mode, with a mass range of 50-1000 Da for MS1 and a collection time of 0.2 s, and a mass range of 25-1000 Da for MS2 and a collection time of 0.04 s. The raw data from the mass spectrometer were converted into mzXM format by ProteoWizard, and the XCMS program was used for peak extraction, alignment, and retention time correction. The peaks with a missing rate of >50% in each group of samples were filtered, and the blank values ​​were filled by KNN, and the peak area was corrected by the SVR method. The peaks after correction and screening were identified by searching the laboratory's self-built database, integrating public libraries, prediction libraries, and metDNA methods for metabolite identification. Finally, substances with a comprehensive score of more than 0.5 and a QC sample CV value of less than 0.5 were extracted and identified, and then the positive and negative modes were merged. Based on the variable importance projection (VIP) obtained by the orthogonal partial least squares discriminant analysis model, differential metabolites were preliminarily screened out, and metabolites with VIP>1 were significantly different. At the same time, combined with the P-value of the univariate analysis, the differential metabolites were further screened out, and P≤0.01 was extremely significant.

[0061] The results are as follows Figure 3Compared with the fruits of the control group that did not overexpress the FaLEC2 gene, the contents of vinblastine, 3-deoxy-D-manno-2-octulose acid, rubiside and linolenic acid in the strawberry fruits of the experimental group that overexpressed the FaLEC2 gene increased significantly, indicating that the FaLEC2 gene can positively regulate the metabolism of vinblastine, 3-deoxy-D-manno-2-octulose acid, rubiside and linolenic acid in strawberry fruits.

[0062] The above description of the disclosed embodiments enables one skilled in the art to implement or use the present invention. Various modifications to these embodiments will be apparent to one skilled in the art, and the general principles defined herein may be implemented in other embodiments without departing from the spirit or scope of the present invention. Therefore, the present invention will not be limited to the embodiments shown herein, but rather to the widest scope consistent with the principles and novel features disclosed herein.

Claims

1. A strawberry FaLEC2 gene, characterized in that: The nucleotide sequence of the FaLEC2 gene is shown in SEQ ID NO.

1.

2. A protein encoded by the strawberry FaLEC2 gene according to claim 1, characterized in that: The amino acid sequence of the protein is shown in SEQ ID NO.

2.

3. Use of the strawberry FaLEC2 gene as claimed in claim 1 in regulating the content of vinblastine, 3-deoxy-D-manno-2-octulose acid, raspberry glycoside or linolenic acid in strawberry fruit.

4. The use of a strawberry FaLEC2 gene as claimed in claim 3 in regulating the content of vinblastine, 3-deoxy-D-manno-2-octulose acid, raspberry glycoside or linolenic acid in strawberry fruit, characterized in that: First, an overexpression vector of the strawberry FaLEC2 gene is constructed; then the overexpression vector is transformed into Agrobacterium; finally, the transformed Agrobacterium is infecting strawberry fruit.

5. The use of a strawberry FaLEC2 gene as claimed in claim 3 in regulating the content of vinblastine, 3-deoxy-D-manno-2-octulose acid, raspberry glycoside or linolenic acid in strawberry fruit, characterized in that: First, an overexpression vector of strawberry FaLEC2 gene is constructed; then the overexpression vector is transformed into Agrobacterium; finally, strawberry tissue culture seedlings are immersed in Agrobacterium bacterial solution carrying FaLEC2 gene for infection, and subcultured to obtain transgenic strawberry plants.

6. Use of a strawberry FaLEC2 gene as claimed in claim 4 or 5 in regulating the content of vinblastine, 3-deoxy-D-manno-2-octulose acid, raspberry glycoside or linolenic acid in strawberry fruit, characterized in that: The overexpression vector is a plasmid containing pC1300s-GFP-FaLEC2.

7. Use of a strawberry FaLEC2 gene as claimed in claim 4 or 5 in regulating the content of vinblastine, 3-deoxy-D-manno-2-octulose acid, raspberry glycoside or linolenic acid in strawberry fruit, characterized in that: The Agrobacterium was GV3101 Agrobacterium.

8. Use of a strawberry FaLEC2 gene as claimed in claim 4 or 5 in regulating the content of vinblastine, 3-deoxy-D-manno-2-octulose acid, raspberry glycoside or linolenic acid in strawberry fruit, characterized in that: The overexpression vector was transformed into Agrobacterium using the freeze-thaw method.

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