Sassafras caryophyllene synthetase gene CpTPS1 and application thereof

Abstract

The invention provides a sassafras caryophyllene synthetase gene CpTPS1 and application thereof, and belongs to the technical field of plant genetic engineering. According to the invention, a safrole caryophyllene synthetase gene CpTPS1 with a nucleotide sequence as shown in SEQ ID NO.1 is obtained and determined from citral type safrole, an expression protein CpTPS1 of the safrole caryophyllene synthetase gene CpTPS1 with an amino acid sequence as shown in SEQ ID NO.2 and a plant expression vector pCambia1300-CpTPS1-Flag / C are further obtained, and the safrole caryophyllene synthetase gene CpTPS1 can be applied to synthesis and preparation of caryophyllene and can also be applied to synthesis and preparation of citral type safrole. The method is also suitable for plant breeding or promotion of plant caryophyllene synthesis. The method is beneficial to promoting breeding of good sassafras varieties, provides new possibility for obtaining caryophyllene, has important scientific significance for promoting research and development of caryophyllene related drugs, and has wide application prospects.

Description

Technical Field

[0001] The present invention belongs to the technical field of plant genetic engineering, and particularly relates to a sassafras-caryophyllene synthase gene. CpTPS1 and its applications. Background Art

[0002] Sassafras ( Camphora parthenoxylon ) is a plant of the genus Cinnamomum in the Lauraceae family. The leaf essential oils of different plants vary significantly in composition, forming a rich chemical type. Among them, citral-type sassafras leaf essential oil, with citral (composed of neral and geranial, with a total content of approximately 50%) as the main component, is widely used in the fields of medicine, daily chemicals, and food, and has broad development prospects. Caryophyllene (β-Caryophyllene), another key component of citral-type sassafras leaf essential oil (content of approximately 10%), has a boiling point close to citral, making it a key factor in the purification process and quality control of citral. In addition, caryophyllene itself has important medicinal value, with various effects such as anti-inflammatory, antioxidant, insect repellent, antitussive, and inhibition of cancer cell growth.

[0003] Caryophyllene is a sesquiterpenoid compound. Studies have shown that members of the terpene synthase (TPS) subfamily b are key genes controlling the synthesis of sesquiterpenoid compounds in plants. Currently, the available genomic data for Camphora serrata is still unclear, and the key to regulating the synthesis of sesquiterpenoid compounds is unclear. TPS The gene has not yet been isolated, and the regulatory mechanism of caryophyllene synthesis in sassafras remains to be explored. Currently, caryophyllene production mainly involves plant extraction, chemical synthesis, and biosynthesis. In this synthesis pathway, the number of identified caryophyllene synthases is still relatively small.

[0004] Therefore, identifying the key regulatory genes for caryophyllene synthesis in citral-type sassafras and deeply analyzing the synthesis pathway of caryophyllene in sassafras will not only help promote the selection and breeding of excellent sassafras varieties, but also have important scientific significance and application value for promoting the research and development of caryophyllene-related drugs. Summary of the Invention

[0005] In view of the above problems existing in the prior art, the present invention aims to provide a sassafras-caryophyllene synthase gene. CpTPS1 The present invention obtains and identifies a sassafras caryophyllene synthase gene from citral-type sassafras. CpTPS1 , and further obtained the sassafras-caryophyllene synthase gene CpTPS1 The expression protein CpTPS1 and the overexpression vector pCambia1300-CpTPS1-Flag / C can not only be used for the synthesis and preparation of caryophyllene, but also be suitable for plant breeding or promoting plant caryophyllene synthesis.

[0006] In the first aspect, the present invention provides a sassafras-caryophyllene synthase gene CpTPS1, the safrolecaryophyllene synthase gene CpTPS1 The nucleotide sequence is shown in SEQ ID NO.1.

[0007] In a second aspect, the present invention provides a sassafras-caryophyllene synthase gene CpTPS1 The expressed protein CpTPS1, the amino acid sequence of which is shown in SEQ ID NO.2.

[0008] The present invention also provides the above-mentioned sassafras-caryophyllene synthase gene CpTPS1 The preparation method of the expression protein CpTPS1 comprises: based on the sassafras caryophyllene synthase gene CpTPS1 A prokaryotic expression vector is constructed, the obtained expression vector is transformed into a host bacterium or cell, and the obtained host bacterium or cell is cultured to express the CpTPS1 protein.

[0009] Furthermore, based on the sassafras caryophyllene synthase gene CpTPS1 The primer pair sequences designed during the construction of the prokaryotic expression vector are shown in SEQ ID NO. 3 and SEQ ID NO. 4, and the plasmid vector used is pET-32a.

[0010] The present invention also provides the above-mentioned sassafras-caryophyllene synthase gene CpTPS1 Application of the expressed protein in the preparation of caryophyllene.

[0011] In a third aspect, the present invention provides a method comprising the above-mentioned safrolecaryophyllene synthase gene CpTPS1 A plant expression vector is constructed using the primer pair shown in SEQ ID NO. 5 and SEQ ID NO. 6 and the pCambia1300-Flag / C plasmid vector. This plant expression vector is pCambia1300-CpTPS1-Flag / C.

[0012] In a fourth aspect, the present invention provides the above-mentioned sassafras-caryophyllene synthase gene CpTPS1 Or the use of the above plant expression vector in plant breeding or promoting plant caryophyllene synthesis.

[0013] Further, the application in plant breeding includes constructing a sassafras caryophyllene synthase gene CpTPS1 The constructed expression vector is transiently transformed into plants to regulate overexpression or inhibit expression to prepare and breed plants with high or low caryophyllene content; the sassafras caryophyllene synthase gene CpTPS1 The expression vectors include pCambia1300-CpTPS1-Flag / C.

[0014] Furthermore, the application in promoting the synthesis of caryophyllene in plants includes constructing an overexpression vector of the sassafras caryophyllene synthase gene CpTPS1, and transiently transforming the constructed overexpression vector into plants to promote the synthesis of caryophyllene in plants; the sassafras caryophyllene synthase gene CpTPS1 is expressed in the plant tissue. CpTPS1 Overexpression vectors include pCambia1300-CpTPS1-Flag / C.

[0015] Beneficial effects of the present invention: (1) The sassafras-caryophyllene synthase gene provided by the present invention CpTPS1 In plant genetic engineering, it is a key gene for caryophyllene synthesis, which can effectively regulate the synthesis of caryophyllene. Its expressed protein is used in the synthesis and preparation of caryophyllene with high yield and important application value. CpTPS1 The substrate farnesyl pyrophosphate (FPP) was added to the expressed protein to detect the catalytic activity of the recombinant protein. The composition detection by gas chromatography-mass spectrometry (GC-MS) showed that 71.23% of the catalytic product was caryophyllene.

[0016] (2) The sassafras-caryophyllene synthase gene provided by the present invention CpTPS1 The plant expression vector pCambia1300-CpTPS1-Flag / C can overexpress and promote the synthesis of caryophyllene in plants, and has good application prospects. Agrobacterium containing the pCambia1300-CpTPS1-Flag / C plant overexpression vector was injected into the recipient tobacco leaves. After 72 hours of transient transformation, CpTPS1 The gene is expressed in large quantities in tobacco leaves. CpTPS1 The gene significantly promoted the synthesis of caryophyllene. BRIEF DESCRIPTION OF THE DRAWINGS

[0017] Figure 1 For PCR cloning CpTPS1 Gene electrophoresis diagram; Lane 1 is DL2000 Marker; Lane 2 is CpTPS1 Gene PCR products; Figure 2 The figure is the SDS-PAGE electrophoresis of the purified His-CpTPS1 recombinant protein; among them, lane 1 is the supernatant after ultrasonic disruption; lane 2 is the precipitate suspension after ultrasonic disruption; lane 3 is the flow-through; lane 4 is the 25 mM imidazole eluent; lane 5 is the 50 mM imidazole eluent; lane 6 is the 75 mM imidazole eluent 1; lane 7 is the 75 mM imidazole eluent 2; lane 8 is the 100 mM imidazole eluent; lane 9 is the 150 mM imidazole eluent 1; lane 10 is the 150 mM imidazole eluent 2; lane 11 is the 200 mM imidazole eluent; lane 12 is the 250 mM imidazole eluent; Figure 3 The product spectrum of the in vitro catalytic reaction of CpTPS1 protein was detected by GC-MS; Figure A shows the caryophyllene standard; Figure B shows the in vitro catalytic product of CpTPS1 protein; Figure C shows the catalytic product of empty protein; Figure 4 transient overexpression CpTPS1 Component profile of essential oil from tobacco leaves. DETAILED DESCRIPTION

[0018] In order to make the purpose, technical solutions and advantages of the embodiments of the present invention clearer, the technical solutions in the embodiments of the present invention will be clearly and completely described below. Obviously, the described embodiments are part of the embodiments of the present invention, not all of the embodiments. Based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without making creative work are within the scope of protection of the present invention. Unless otherwise defined, the technical terms or scientific terms used herein should be the common meanings understood by people with ordinary skills in the field to which the present invention belongs.

[0019] In the following examples, operations not described in detail are all routine biological experimental operations, which can be performed with reference to molecular biology experimental manuals and existing published journals. Example 1

[0020] Example 1 of the present invention provides a step for obtaining a candidate gene encoding caryophyllene synthase, starting from sample collection and sequencing, screening the target gene through data analysis, extracting sample RNA and quality inspection after designing primers, synthesizing cDNA, amplifying and cloning the target gene, and finally confirming the candidate gene sequence through sequencing result analysis, specifically including: 1. Sample Collection and Sequencing: Young leaf tissues of Citral-type Sassafras were collected and sent to BGI and Shanghai Meiji Biotechnology for full-length transcriptome and next-generation transcriptome sequencing on the PacBio Sequel platform and Illumina NovaSeq 6000 platform.

[0021] 2. Data analysis and gene screening: 8.93 Gb and 7.77 Gb of filtered data were obtained for the full-length transcriptome and the second-generation transcriptome, respectively. Through bioinformatics annotation, 12 TPS subfamily b members were identified; among them, CpTPS1 The gene with the highest expression level was screened as a candidate gene encoding caryophyllene synthase.

[0022] 3. Using the full-length transcriptome data as a reference, design the CpTPS1 Gene cloning primers, the primer pair sequences are shown in SEQ ID NO.7 and SEQ ID NO.8; SEQ ID NO.7 (CpTPS1-F): 5'-CTACTTCCAGCTTACCCCTT-3', SEQ ID NO. 8 (CpTPS1-R): 5'-AATCCCTAGATAAAATGGCAC-3'.

[0023] 4. RNA Extraction and Quality Testing: Total RNA was extracted from young citral-type sassafras leaf tissue using the Polysaccharide and Polyphenol Plant RNA Extraction Kit (ZH120) produced by Beijing Huayueyang Biotechnology Co., Ltd. The quality and concentration of the extracted total RNA were determined by agarose gel electrophoresis and spectrophotometry to ensure that the RNA sample met the requirements of subsequent experiments.

[0024] 5. cDNA synthesis, gene amplification and cloning: 100 ng of qualified total RNA was taken and reverse transcription reaction was performed using the EasyScript All-in-One First-Strand cDNA Synthesis Kit produced by Beijing Quanshijin Biotechnology Co., Ltd. to synthesize cDNA. The high-fidelity enzyme PrimeSTAR produced by TAKARA was used for PCR amplification. CpTPS1 The full-length coding region (CDS) sequence of the gene (estimated length 1815 bp) was detected by agarose gel electrophoresis and the target band was recovered by gel cutting ( Figure 1 ) and inserted into the pEASY®-Blunt cloning vector. Positive single clones were selected, plasmids were extracted, and the clones were sent to Sangon (Shanghai) Biotechnology Co., Ltd. for next-generation sequencing.

[0025] 6. Analysis of sequencing results: Sequencing results showed that the candidate genes CpTPS1 The CDS sequence is 1689 bp and is expected to encode a polypeptide consisting of 562 amino acid residues. Example 2

[0026] Example 2 of the present invention provides a sassafras-caryophyllene synthase gene CpTPS1 To obtain the expressed protein CpTPS1, a His tag is added to the target protein to facilitate subsequent protein purification and analysis, resulting in a His-CpTPS1 fusion protein. The target gene fragment is amplified by primer design and PCR, and then constructed into the prokaryotic expression vector pET-32a (which contains a histidine His tag) to form a recombinant plasmid. After transformation into the host bacteria and induction of expression, protein expression is initially tested, followed by purification and SDS-PAGE electrophoresis verification. Finally, the protein concentration is measured to obtain a highly pure fusion protein. The steps include: 1. Primer design and PCR amplification: Primers for constructing the prokaryotic expression vector were designed according to the instructions of the ClonExpress II One Step Cloning Kit (C112) produced by Nanjing Novozymes. The primer pair sequences are shown in SEQ ID NO. 3 and SEQ ID NO. 4. SEQ ID NO.3 (5P): 5'-gctgatatcggatccgaattcaATGGCTCTTGTTTCTGGTTC-3', SEQ ID NO.4 (3P): 5'-tgcggccgcaagcttgtcgacTCAAATTGGGATAGGATT-3'.

[0027] Using the pEASY®-Blunt-CpTPS1 dilution as a template, amplify the DNA using the high-fidelity enzyme PrimeSTAR. CpTPS1 The CDS sequence of the gene was detected by agarose gel electrophoresis and the target product was recovered by gel excision.

[0028] 2. Construction of the prokaryotic expression vector pET-32a-CpTPS1: First, the pET-32a plasmid was double-digested with the endonucleases EcoR I and Sal I to recover the linearized plasmid backbone DNA. Subsequently, an enzymatic reaction solution (2 μL of 5× CE II Buffer, 50 ng of linearized plasmid DNA, 1 μL of Exnase, 100 ng of CpTPS1-DNA, and supplemented with ddH2O to 10 μL) was prepared and incubated at 37°C for 30 min. Finally, the reaction product was transformed into competent Escherichia coli Top10 cells using the heat shock method, plated onto LB solid medium supplemented with 100 mg / L ampicillin, and incubated inverted at 37°C for 12–16 h. Single colonies were picked, and positive clones were screened by PCR and confirmed by Sanger sequencing to confirm the successful construction of the pET-32a-CpTPS1 vector.

[0029] 4. Recombinant plasmid transformation and protein induction expression: Extract the recombinant plasmid pET-32a-CpTPS1 and transform it into the protein induction expression strain BL21 (DE3) competent cells by heat shock method. Spread it on LB solid medium containing 100 mg / L ampicillin and culture at 37°C for 12-16 hours. Screen the positive single clones and inoculate them into 2 mL LB liquid medium containing the same antibiotics and culture overnight. The next day, take 1 mL of the bacterial solution and transfer it to 50 mL LB liquid medium containing the same antibiotics for amplification. Wait until the bacterial solution OD 600When the value is approximately 0.8, add the protein inducer IPTG (isopropyl-β-D-thiogalactopyranoside) to a final concentration of 0.5 mM and culture with shaking at 20°C for approximately 16 h. Set up a blank control by transforming the empty vector pET-32a.

[0030] 5. Preliminary protein expression testing: 2 mL of each bacterial suspension was centrifuged at 12,000 rpm for 10 min to collect the cells. The pellet was resuspended in 200 μL of PBS buffer and boiled for 10 min to lyse the cells. The pellet was centrifuged at 12,000 rpm for 10 min and the supernatant collected. The pellet was resuspended in 100 μL of PBS buffer. 10 μL of each supernatant and pellet suspension were added to 2× protein loading buffer and analyzed by SDS-PAGE electrophoresis.

[0031] 6. Protein Purification and Verification: After confirming that the His-CpTPS1 protein was successfully expressed in the supernatant, the remaining cells were collected by centrifugation and resuspended in 5 mL of PBS buffer. The cells were ultrasonically disrupted on ice and subjected to refrigerated ultracentrifugation (12,000 rpm, 4°C) for 15 min. The His-CpTPS1 recombinant protein was purified from the supernatant using a Ni Sepharose FF kit produced by Nanjing Novozymes, and the purified product was detected by SDS-PAGE electrophoresis (see ). Figure 2 The total size of the tagged protein and CpTPS1 was estimated to be approximately 85 kDa, consistent with the electrophoresis results.

[0032] 7. Protein concentration determination: The recombinant protein concentration was determined using the BCA protein concentration test kit produced by Yisheng Biotechnology. The specific operation was carried out according to the instructions. Example 3

[0033] In vitro catalytic reaction verification of Example 3 of the present invention CpTPS1 Gene function, mainly includes constructing an in vitro catalytic reaction system, extracting the reaction products, and using GC-MS technology to detect and analyze the product components to verify CpTPS1 The functions of the genes, the specific steps and the results are as follows: 1. In vitro catalytic reaction: In 50 mmol / L Bis-Tris (pH 7.2) buffer, add 1 μg of His-CpTPS1 recombinant protein, 10 μmol / L farnesyl pyrophosphate (FPP, purchased from Sigma-Aldrich), 10 mM MgCl₂, 10 mM MnCl₂, 10% glycerol, and 5 mM DTT to prepare a reaction solution. Place the solution in a 5 mL crimp-top vial (total reaction volume 0.5 mL). Seal the vial and incubate at 30°C in a shaker with slow shaking for 3 h. Simultaneously, an equal amount of empty protein was added as a control for in vitro catalytic experiments using the same system and conditions.

[0034] 2. Product extraction: After the reaction, a solid phase microextraction fiber probe (Supelco 100 μm PDMS, Fused Silica 24Ga, Manual Holder, 3pk (Red)) was inserted into the crimp-top bottle and the product was extracted at 60°C for 30 min.

[0035] 3. GC-MS detection of product components: Shimadzu gas chromatography-mass spectrometry (GC-MS) was used to analyze the product components. The specific conditions are as follows: Carrier gas: helium, flow rate 1.0 mL / min, split injection, split ratio 20:1; gas phase conditions (GC): inlet temperature 280°C; temperature program: initial column temperature 50°C for 2 min, temperature increase at 3°C / min to 180°C for 2 min, then temperature increase at 8°C / min to 240°C for 5 min, total run time 60 min; mass spectrometry conditions (MS): interface temperature 260°C, ion source temperature 180°C, scan range (m / z) 50-620.

[0036] Data analysis: The product components were matched against the NIST standard spectral library, and components with a matching score higher than 85 were screened out. The components were then compared with the chromatogram of a standard caryophyllene (purchased from Sigma-Aldrich), and the relative content of the products was determined by the peak area ratio.

[0037] 4. Result analysis: Figure 3 As shown in the results, when FPP was used as substrate, the catalytic products of CpTPS1 protein mainly included caryophyllene (71.23%) and α-Caryophyllene (28.77%), confirming that CpTPS1 protein has the function of synthesizing caryophyllene. Example 4

[0038] Example 4 of the present invention Nicotiana benthamiana ( Nicotiana benthamiana ) was used in a transient overexpression experiment. The main steps involved designing primers and amplifying the target gene fragment through PCR. The overexpression vector was constructed using the pCambia1300-Flag / C plasmid vector (which contains a Flag tag at the C-terminus of the inserted gene). After transformation with Agrobacterium tumefaciens GV3101, an infection solution was prepared and infiltrated into tobacco leaves. Finally, essential oil from the leaves was extracted and analyzed for composition to verify gene function. The specific steps are as follows: 1. Primer design and PCR amplification: Referring to the instructions of the cloning kit exv06 produced by Biogen, primers for constructing the overexpression vector pCambia1300-CpTPS1-Flag / C were designed. The primer pair sequences are shown in SEQ ID NO. 5 and SEQ ID NO. 6. SEQ ID NO.5 (5P): 5'-tcagcagtcgaagagcATGGCTCTTGTTTCTGGTTC-3', SEQ ID NO.6 (3P): 5'-ttagcgtgtgaagagcTCAAATTGGGATAGGATTA-3'.

[0039] The cloning plasmid pEASY®-Blunt-CpTPS1 was used as a template and amplified using the high-fidelity enzyme PrimeSTAR. CpTPS1 The CDS sequence of the gene was detected by agarose gel electrophoresis, and the CpTPS1-DNA was recovered by gel excision.

[0040] 2. Construction of the overexpression vector pCambia1300-CpTPS1-Flag / C: First, prepare the enzymatic reaction system (2 μL of 5× EX II Buffer, 1 μL of Exclonase Enzyme, 50 ng of linearized pCambia1300-Flag / C vector DNA, 100 ng of CpTPS1-DNA, and add ddH2O to 10 μL). Incubate the reaction at 37°C for 30 min and then incubate at room temperature for 15 min. The reaction product was then transformed into competent Escherichia coli Top10 cells using the heat shock method, plated onto LB solid medium supplemented with 50 mg / L kanamycin, and incubated at 37°C for 12–16 h. Finally, positive single colonies were screened by PCR, and successful vector construction was confirmed by Sanger sequencing.

[0041] 3. Transformation of recombinant plasmid into Agrobacterium tumefaciens: Extract the recombinant plasmid pCambia1300-CpTPS1-Flag / C and transform it into Agrobacterium tumefaciens GV3101 competent cells by heat shock method. Spread it on LB solid medium containing 50 mg / L kanamycin, 100 mg / mL rifampicin, and 50 mg / L gentamicin, and culture at 37°C for 12-16 hours. Select positive single colonies and inoculate them into 50 mL LB liquid medium containing the same antibiotics. Culture at 37°C until the OD value of the bacterial solution reaches 0. 600 The value was about 0.8. The cells were collected by centrifugation and rinsed twice with 10 mM MgCl2 to completely remove the residual rifampicin.

[0042] 4. Prepare infection solution and infect tobacco leaves: Prepare infection solution (200 mM acetosyringone, 0.5 mM MES and 10 mM MgCl2), suspend the bacteria and adjust the bacterial concentration OD 600Set the value to approximately 1.0 and incubate at room temperature in the dark for 2-3 hours. Take a growing Nicotiana benthamiana leaf and poke several small holes on the underside of the leaf with a needle. Place the infection solution into a 1 mL syringe and inject the solution through the lower epidermis of the leaf. The leaf surface will become moist after injection. As a control, inoculate with GV3101 bacteria containing the empty vector pCambia1300-Flag / C. Incubate in the dark for 1 day and under normal light for 2 days.

[0043] 5. Essential oil extraction and component analysis: 1 g of sample was ground into powder and 10 mL of n-hexane was added to extract the essential oil for 3 h. Centrifuge at 12,000 rpm for 30 min, collect the supernatant, and filter 0.22 μL of organic film. The essential oil components were determined by GC-MS under the same conditions as in Example 3. The results are shown in Figure 2. Figure 4 Showing that transient overexpression CpTPS1 After the gene was added, the contents of caryophyllene and humulene in tobacco samples were significantly increased compared with the control.

[0044] While the embodiments of the present invention have been described in detail above, it will be apparent to those skilled in the art that various modifications and variations of these embodiments are possible. However, it should be understood that such modifications and variations are within the scope and spirit of the present invention as set forth in the claims. Furthermore, the invention described herein is susceptible to other embodiments and may be practiced or implemented in a variety of ways.

Claims

1. A sassafras-caryophyllene synthase gene CpTPS1 , characterized in that, The safrolecaryophyllene synthase gene CpTPS1 The nucleotide sequence is shown in SEQ ID NO.

1.

2. A sassafras-caryophyllene synthase gene CpTPS1 The expression protein CpTPS1 is characterized in that The amino acid sequence of the protein is shown in SEQ ID NO.

2.

3. A method for preparing the protein according to claim 2, characterized in that: Including: based on safrole caryophyllene synthase gene CpTPS1 A prokaryotic expression vector is constructed, the obtained expression vector is transformed into a host bacterium or cell, and the obtained host bacterium or cell is cultured to express the CpTPS1 protein.

4. The preparation method according to claim 3, characterized in that The prokaryotic expression vector is constructed using the primer pair shown in SEQ ID NO. 3 and SEQ ID NO. 4 and the pET-32a plasmid vector.

5. Use of the protein according to claim 2 or the protein prepared by the method according to any one of claims 3 to 4 in the preparation of caryophyllene.

6. A plant expression vector comprising the gene according to claim 1, characterized in that: The plant expression vector is constructed using the primer pair shown in SEQ ID NO. 5 and SEQ ID NO. 6 and the pCambia1300-Flag / C plasmid vector.

7. A sassafras-caryophyllene synthase gene as claimed in claim 1 CpTPS1 Or use of the plant expression vector according to claim 6 in plant breeding or promoting caryophyllene synthesis in plants.

8. The use according to claim 7, characterized in that The application in plant breeding comprises constructing a sassafras caryophyllene synthase gene CpTPS1 The constructed expression vector is transiently transformed into plants to regulate overexpression or inhibit expression to prepare and breed plants with high or low caryophyllene content; the sassafras caryophyllene synthase gene CpTPS1 The expression vector includes the plant expression vector according to claim 6.

9. The use according to claim 7, characterized in that The application in promoting the synthesis of plant caryophyllene comprises constructing a sassafras caryophyllene synthase gene CpTPS1 The overexpression vector is transiently transformed into plants to promote the synthesis of caryophyllene in plants; the sassafras caryophyllene synthase gene CpTPS1 The overexpression vector comprises the plant expression vector according to claim 6.

Images