CN107189976B

CN107189976B - Method for promoting expression of momordica grosvenori UGT72B21 gene

Info

Publication number: CN107189976B
Application number: CN201710630164.6A
Authority: CN
Inventors: 李华政; 韦荣昌
Original assignee: Individual
Current assignee: Individual
Priority date: 2017-07-28
Filing date: 2017-07-28
Publication date: 2022-03-11
Anticipated expiration: 2037-07-28
Also published as: CN107189976A

Abstract

The invention discloses a method for promoting expression of momordica grosvenori UGT72B21 gene, comprising the following steps: in the tissue culture process and the cultivation process of the momordica grosvenori, methyl jasmonate with the concentration of 50-400 mu mol/L is applied to promote the expression of the momordica grosvenori UGT72B21 gene. According to the invention, methyl jasmonate is applied to the tissue culture seedlings of the momordica grosvenori and the cultivated momordica grosvenori, so that the high expression of a key enzyme gene UGT72B21 in the biosynthesis pathway of the momordica grosvenori sweet glycoside V is quickly induced in a short time, and a solid foundation is laid for promoting the accumulation of the content of the momordica grosvenori sweet glycoside V; the method is simple and convenient to operate, low in cost, environment-friendly, suitable for large-scale production and high in practicability and popularization value.

Description

Method for promoting expression of momordica grosvenori UGT72B21 gene

Technical Field

The invention belongs to the technical field of plant biology, and relates to a method for promoting expression of momordica grosvenori UGT72B21 gene.

Background

Mogroside V belongs to cucurbitane tetracyclic triterpenoids, and the applicant has deduced a possible biosynthetic pathway of mogroside V according to mogroside transcriptome data in the early stage. Precursors for mogroside V biosynthesis are isopentenyl diphosphate (IPP) and 3, 3-dimethylenemesityl pyrophosphate (DMAPP), which are formed via both Mevalonate (MVA) and methylerythritol phosphorylation (MEP), the MVA pathway occurring in the cytoplasm and the MEP pathway occurring in the plastids. IPP or DMAPP from the two ways is catalyzed by Geranyl Pyrophosphate Synthase (GPS) to form geranyl pyrophosphate (GPP), the IPP and GPP further form farnesyl pyrophosphate (FPP) under the catalysis of Farnesyl Pyrophosphate Synthase (FPS), then form 2, 3-oxidosqualene by the catalysis of Squalene Synthase (SS) and Squalene Epoxidase (SE), the Cucurbitadienol Synthase (CS) further catalyzes to form cucurbitadienol, and finally form mogroside V under the action of CYP450 enzyme and glucosyltransferase (UGT).

The production of isoprenoids is strictly regulated by the activity of rate-limiting enzymes, and has been considered to play an important role in the biosynthesis pathway. As a rate-limiting enzyme in the isoprene pathway, the expression of UGT gene plays a decisive role in the biosynthesis of mogroside V. The UGT gene is overexpressed, so that the accumulation of mogroside V can be promoted; in contrast, if the expression of UGT gene is suppressed, the production of mogroside V will be significantly reduced. However, UGT gene is a super gene family, and the applicant finds that UGT72B21 may be involved in the synthetic pathway of mogroside V in earlier researches. No report for promoting the expression of the momordica grosvenori UGT72B21 gene exists in the prior art.

Disclosure of Invention

An object of the present invention is to solve at least the above problems and/or disadvantages and to provide at least the advantages described hereinafter.

The invention also aims to provide a method for promoting the expression of the UGT72B21 gene of the momordica grosvenori, which has the characteristics of quick response, low cost, simple operation, convenient implementation and the like and lays a foundation for effectively promoting the accumulation of mogroside V.

Therefore, the technical scheme provided by the invention is as follows:

a method for promoting expression of momordica grosvenori UGT72B21 gene comprises the following steps: in the tissue culture process and the cultivation process of the momordica grosvenori, methyl jasmonate with the concentration of 50-400 mu mol/L is applied to promote the expression of the momordica grosvenori UGT72B21 gene.

Preferably, in the method for promoting expression of momordica grosvenori UGT72B21 gene, the methyl jasmonate is added to the solid culture medium in the momordica grosvenori tissue culture process.

Preferably, in the method for promoting the UGT72B21 gene expression of the momordica grosvenori, the methyl jasmonate with the concentration of 50-400 mu mol/L is sprayed on the surface of the momordica grosvenori 20-30 days after pollination until water drops on the surface of the momordica grosvenori, and the methyl jasmonate is sprayed once in the morning, at noon and at night every day and is continuously sprayed for 5 days.

Preferably, in the method for promoting expression of momordica grosvenori UGT72B21 gene, the solid medium in the momordica grosvenori tissue culture process comprises: MS, 1.5mg/L6-BA, 0.3mg/L IBA, 3.5g/L agar, 30g/L sucrose and 1.0g/L activated carbon.

Preferably, in the method for promoting expression of momordica grosvenori UGT72B21 gene, the culture conditions in the momordica grosvenori tissue culture process are as follows: culturing for 30 days under the conditions of relative humidity of 60-66%, illumination intensity of 1400lux, illumination time of 8h/d and temperature of 21-25 ℃.

Preferably, the method for promoting expression of momordica grosvenori UGT72B21 gene further comprises: preparing a jasmonic acid mother solution: dissolving methyl jasmonate in 2% (v/v) ethanol water solution to prepare 10000 mu mol/L mother liquor, and sterilizing with a 0.22 mu m microporous filter membrane to obtain sterilized methyl jasmonate mother liquor;

adding the methyl jasmonate mother liquor into a solid culture medium in the tissue culture process of the momordica grosvenori till the final concentration of the methyl jasmonate is 50-400 mu mol/L;

dissolving with ethanol, preparing 50-400 μmol/L solution from methyl jasmonate mother liquor, and applying to the fructus Siraitiae Grosvenorii cultivation process.

Preferably, the method for promoting expression of momordica grosvenori UGT72B21 gene further comprises the following steps:

qRT-PCR detection of UGT72B21 Gene expression:

1) collecting fructus Siraitiae Grosvenorii fruit, cutting into 2-4mm pieces, wrapping with tinfoil paper, quickly freezing in liquid nitrogen, and storing at-80 deg.C;

2) extracting total RNA of fructus momordicae fruits by adopting a Trizol method;

3) reverse transcribing the RNA to cDNA;

4) detecting the expression quantity of UGT72B21 gene by adopting ABI7500 real-time fluorescent quantitative PCR instrument, wherein the primer sequence for amplifying UGT72B21 is as follows: an upstream primer CGCCATATGCACCACCACCACCACCACATGGAAGCTACGCAGAGAGACG, a downstream primer CCCTCGAGTTAAGACTCAAGAAATGCAAACTTA and a UBQ5 gene for an internal reference gene, wherein the sequence of an amplification primer is as follows: an upstream primer ATAAAAGACCCAGCACCACATTC and a downstream primer CCCTTGCCGACTACAACATCC.

Preferably, in the method for promoting the UGT72B21 gene expression of the momordica grosvenori, the temperature of methyl jasmonate sprayed on the surface of the momordica grosvenori is 4-10 ℃ in the cultivation process of the momordica grosvenori, and meanwhile, the methyl jasmonate is injected into a main stem of a momordica grosvenori plant in an injection mode, the injection amount is 5-16 ml, and the injection speed is 4-8 ml/h.

The invention at least comprises the following beneficial effects:

(1) according to the invention, methyl jasmonate is applied to the tissue culture seedlings of the momordica grosvenori and the cultivated momordica grosvenori, so that the high expression of a key enzyme gene UGT72B21 in the biosynthesis pathway of the momordica grosvenori sweet glycoside V is quickly induced in a short time, and a solid foundation is laid for promoting the accumulation of the content of the momordica grosvenori sweet glycoside V;

(2) the method is simple and convenient to operate, low in cost, environment-friendly, suitable for large-scale production and high in practicability and popularization value.

Additional advantages, objects, and features of the invention will be set forth in part in the description which follows and in part will become apparent to those having ordinary skill in the art upon examination of the following or may be learned from practice of the invention.

Detailed Description

The present invention is further described in detail below with reference to examples so that those skilled in the art can practice the invention with reference to the description.

It will be understood that terms such as "having," "including," and "comprising," as used herein, do not preclude the presence or addition of one or more other elements or groups thereof.

The invention provides a method for promoting expression of momordica grosvenori UGT72B21 gene, comprising the following steps: in the tissue culture process and the cultivation process of the momordica grosvenori, methyl jasmonate with the concentration of 50-400 mu mol/L is applied to promote the expression of the momordica grosvenori UGT72B21 gene.

In one embodiment of the present invention, preferably, during the grosvenor momordica tissue culture process, the methyl jasmonate is added to a solid medium during the grosvenor momordica tissue culture process.

In the above scheme, preferably, the solid medium in the process of tissue culture of luo han guo comprises: MS, 1.5mg/L6-BA, 0.3mg/L IBA, 3.5g/L agar, 30g/L sucrose and 1.0g/L activated carbon.

In one embodiment of the invention, preferably, in the cultivation process of the momordica grosvenori, after 20-30 days after pollination of the momordica grosvenori, the methyl jasmonate with the concentration of 50-400 mu mol/L is sprayed on the surface of the momordica grosvenori until water drops on the surface of the momordica grosvenori, and the methyl jasmonate is sprayed once in the morning, at noon and at night every day and is continuously sprayed for 5 days.

In one embodiment of the present invention, preferably, the culture conditions during the tissue culture of luo han guo are: culturing for 30 days under the conditions of relative humidity of 60-66%, illumination intensity of 1400lux, illumination time of 8h/d and temperature of 21-25 ℃.

In one embodiment of the present invention, it is preferable that: preparing a jasmonic acid mother solution: dissolving methyl jasmonate in 2% (v/v) ethanol water solution to prepare 10000 mu mol/L mother liquor, and sterilizing with a 0.22 mu m microporous filter membrane to obtain sterilized methyl jasmonate mother liquor;

In one embodiment of the present invention, it is preferable that the method further includes the following steps:

qRT-PCR detection of UGT72B21 Gene expression:

3) reverse transcribing the RNA to cDNA;

4) detecting the expression quantity of UGT72B21 gene by adopting ABI7500 real-time fluorescent quantitative PCR instrument, wherein the primer sequence for amplifying UGT72B21 is as follows: an upstream primer CGCCATATGCACCACCACCACCACCACATGGAAGCTACGCAGAGAGACG (SEQ ID NO:1), a downstream primer CCCTCGAGTTAAGACTCAAGAAATGCAAACTTA (SEQ ID NO:2), a UBQ5 gene for an internal reference gene, and an amplification primer sequence of: an upstream primer ATAAAAGACCCAGCACCACATTC (SEQ ID NO:3), a downstream primer CCCTTGCCGACTACAACATCC (SEQ ID NO: 4).

In one embodiment of the invention, preferably, in the cultivation process of the momordica grosvenori, the temperature of the methyl jasmonate sprayed on the surface of the momordica grosvenori is 4-10 ℃, and meanwhile, the methyl jasmonate is injected into a main stem of the momordica grosvenori plant in an injection mode, the injection amount is 5-16 ml, and the injection speed is 4-8 ml/h. In the cultivation process of the momordica grosvenori, the methyl jasmonate solution at the temperature is sprayed on the surface of the momordica grosvenori, certain cold stimulation is given to fruits of the momordica grosvenori, and the UGT72B21 gene expression can be further induced. The methyl jasmonate is directly injected into a grosvenor momordica body, so that direct absorption of grosvenor momordica plants can be facilitated, the absorption efficiency is further improved, and cold stimulation is more obvious due to the fact that the methyl jasmonate is directly injected into the body, and the expression of UGT72B21 gene can be further promoted.

Example 1

A method for promoting expression of momordica grosvenori UGT72B21 gene, comprising the following steps:

(1) applying methyl jasmonate to the grosvenor momordica tissue culture seedlings; dissolving methyl jasmonate in 2% ethanol water solution (v/v) to prepare 10000 mu mol/L mother solution, sterilizing the mother solution by using a 0.22 mu m microporous filter membrane, adding the sterilized methyl jasmonate mother solution into a solid culture medium until the final concentration of the methyl jasmonate is 50 mu mol/L, sterilizing and cooling to 24 ℃; the proportion of the solid culture medium is MS +6-BA1.5mg/L + IBA0.3mg/L +3.5g/L agar +30g/L cane sugar +1.0g/L active carbon. Inoculating the tissue culture seedling of the momordica grosvenori into a solid culture medium containing methyl jasmonate, and culturing for 30d under the conditions of relative humidity of 60%, illumination intensity of 1400lux, illumination time of 8h/d and temperature of 23 ℃.

(2) Cultivating fructus Siraitiae Grosvenorii and adding methyl jasmonate; dissolving with small amount of ethanol, preparing 50 μmol/L solution of methyl jasmonate stock solution, spraying onto surface of fructus Siraitiae Grosvenorii 20d after pollination, spraying once in the morning, in the evening, and continuously spraying for 5 d.

While comparative example 1 was set up,

1) inoculating the grosvenor momordica tissue culture seedling into a solid culture medium which does not contain methyl jasmonate for culture;

2) the methyl jasmonate is not sprayed on the momordica grosvenori plants; the other conditions were the same as in example 1.

(3) qRT-PCR detects the expression of UGT72B21 gene.

Collecting fruit, picking pulp, cutting into 2mm pieces, wrapping with tinfoil paper, quickly freezing in liquid nitrogen, and storing at-80 deg.C.

Extracting total RNA of fructus Siraitiae Grosvenorii fruit by improved Trizol method.

An ABI7500 real-time fluorescence quantitative PCR instrument is adopted.

The reaction system for reverse transcription of RNA into cDNA was RNA 10.0. mu.L, PrimeScript RT Enzyme Mix I1.0. mu.L, RT Primer Mix 1.0. mu.L, 5 XPrimeScript Buffer 24.0. mu.L, RNase Free dH₂O 4.0μL; the reaction conditions were 37 ℃ (15min) → 85 ℃ (5s) → 4 ℃.

Primers were designed using Primer Premier 5.0 software and synthesized by Biotechnology engineering (Shanghai) GmbH. Wherein the primer sequence of UGT72B21 is (FP: CGCCATATGCACCACCACCACCACCACATGGAAGCTACGCAGAGAGACG, RP: CCCTCGAGTTAAGACTCAAGAAATGCAAACTTA), the sequence of UBQ5 for the reference gene is (FP: ATAAAAGACCCAGCACCACATTC, RP: CCCTTGCCGACTACAACATCC).

The qRT-PCR reaction system (20. mu.L) was SYBR Premix Ex Taq II (Tli RNaseH Plus) (2X) 10.0. mu.L, PCR Forward Primer (10. mu.M) 0.8. mu.L, PCR Reverse Primer (10. mu.M) 0.8. mu.L, ROX Reference Dye of Dye II (50X) 0.4. mu.L, Template 2.0. mu.L, dH₂O6.0. mu.L. The reaction conditions were 95 deg.C (30s), followed by 40 cycles [95 deg.C (5s), 95 deg.C (34s)]。

The qRT-PCR detection result shows that the expression level of the momordica grosvenori UGT72B21 gene in the comparative example 1 is 1, while the expression level of the momordica grosvenori UGT72B21 gene in the example 1 is 7.3, which is improved by 630% compared with the comparative example 1, and thus, the example 1 can remarkably promote the high expression of the momordica grosvenori UGT72B21 gene; HPLC detection results show that in comparative example 1, the mogroside V content is 0.80%, while in example 1, the mogroside V content is as high as 1.60%, which is 100% higher than that in comparative example 1, and it can be seen that in example 1, the mogroside V yield can be remarkably improved.

Example 2

(1) applying methyl jasmonate to the grosvenor momordica tissue culture seedlings; dissolving methyl jasmonate in 2% ethanol water solution (v/v) to prepare 10000 mu mol/L mother solution, sterilizing the mother solution by using a 0.22 mu m microporous filter membrane, adding the sterilized methyl jasmonate mother solution into a solid culture medium until the final concentration of the methyl jasmonate is 400 mu mol/L, sterilizing and cooling to 26 ℃; the proportion of the solid culture medium is MS +6-BA1.5mg/L + IBA0.3mg/L +3.5g/L agar +30g/L cane sugar +1.0g/L active carbon. Inoculating the Momordica grosvenori tissue culture seedling in a solid culture medium containing methyl jasmonate, and culturing at 21 deg.C for 30d under the conditions of relative humidity of 66%, illumination intensity of 1400lux and illumination time of 8 h/d.

(2) Cultivating fructus Siraitiae Grosvenorii and adding methyl jasmonate; dissolving with small amount of ethanol, preparing 400 μmol/L solution of methyl jasmonate stock solution, spraying onto the surface of fructus Siraitiae Grosvenorii 30d after pollination, spraying once in the morning, in the evening and continuously for 5 d.

While the comparative example 2 was set up,

(3) qRT-PCR detects the expression of UGT72B21 gene.

Collecting fruit, picking pulp, cutting into 4mm small pieces, wrapping with tinfoil paper, quickly freezing in liquid nitrogen, and storing at-80 deg.C.

An ABI7500 real-time fluorescence quantitative PCR instrument is adopted.

The reaction system for reverse transcription of RNA into cDNA was RNA 10.0. mu.L, PrimeScript RT Enzyme Mix I1.0. mu.L, RT Primer Mix 1.0. mu.L, 5 XPrimeScript Buffer 24.0. mu.L, RNase Free dH₂O4.0 μ L; the reaction conditions were 37 ℃ (15min) → 85 ℃ (5s) → 4 ℃.

The qRT-PCR detection result shows that the expression level of the momordica grosvenori UGT72B21 gene in the comparative example 2 is 1, while the expression level of the momordica grosvenori UGT72B21 gene in the example 2 is 10.3, which is improved by 930% compared with the comparative example 2, and therefore, the example 2 can obviously promote the high expression of the momordica grosvenori UGT72B21 gene; HPLC detection results show that in comparative example 2, the mogroside V content is 0.80%, while in example 2, the mogroside V content is as high as 2.43%, which is 203.75% higher than that in comparative example 2, and it can be seen that in example 2, the mogroside V yield can be significantly improved.

Example 3

(1) applying methyl jasmonate to the grosvenor momordica tissue culture seedlings; dissolving methyl jasmonate in 2% ethanol water solution (v/v) to prepare 10000 mu mol/L mother liquor, sterilizing the mother liquor by using a 0.22 mu m microporous filter membrane, adding the sterilized methyl jasmonate mother liquor into a solid culture medium until the final concentration of the methyl jasmonate is 225 mu mol/L, sterilizing and cooling to 24-26 ℃; the proportion of the solid culture medium is MS +6-BA1.5mg/L + IBA0.3mg/L +3.5g/L agar +30g/L cane sugar +1.0g/L active carbon. Inoculating the tissue culture seedling of the momordica grosvenori into a solid culture medium containing methyl jasmonate, and culturing for 30d under the conditions of relative humidity of 63%, illumination intensity of 1400lux, illumination time of 8h/d and temperature of 25 ℃.

(2) Cultivating fructus Siraitiae Grosvenorii and adding methyl jasmonate; dissolving with small amount of ethanol, preparing 225 μmol/L methyl jasmonate stock solution, spraying onto the surface of fructus Siraitiae Grosvenorii 25d after pollination, spraying once in the morning, in the evening, and continuously spraying for 5 d.

While comparative example 3 was set up with the addition of,

(3) qRT-PCR detects the expression of UGT72B21 gene.

Collecting fruit, picking pulp, cutting into 3mm small pieces, wrapping with tinfoil paper, quickly freezing in liquid nitrogen, and storing at-80 deg.C.

An ABI7500 real-time fluorescence quantitative PCR instrument is adopted.

The qRT-PCR detection result shows that the expression level of the momordica grosvenori UGT72B21 gene in the comparative example 3 is 1, while the expression level of the momordica grosvenori UGT72B21 gene in the example 3 is up to 18.7, which is improved by 1770% compared with the comparative example 3, and thus, the example 3 can obviously promote the high expression of the momordica grosvenori UGT72B21 gene; HPLC detection results show that in comparative example 3, the mogroside V content is 0.80%, while in example 3, the mogroside V content is as high as 2.96%, which is improved by 270% compared with comparative example 3, and thus, in example 3, the mogroside V yield can be improved remarkably.

Example 4

(1) applying methyl jasmonate to the grosvenor momordica tissue culture seedlings; dissolving methyl jasmonate in 2% ethanol water solution (v/v) to prepare 10000 mu mol/L mother solution, sterilizing the mother solution by using a 0.22 mu m microporous filter membrane, adding the sterilized methyl jasmonate mother solution into a solid culture medium until the final concentration of the methyl jasmonate is 100 mu mol/L, sterilizing and cooling to 24-26 ℃; the proportion of the solid culture medium is MS +6-BA1.5mg/L + IBA0.3mg/L +3.5g/L agar +30g/L cane sugar +1.0g/L active carbon. Inoculating the Momordica grosvenori tissue culture seedling in a solid culture medium containing methyl jasmonate, and culturing at 24 deg.C for 30d under the conditions of relative humidity of 64%, illumination intensity of 1400lux, and illumination time of 8 h/d.

(2) Cultivating fructus Siraitiae Grosvenorii and adding methyl jasmonate; dissolving with a small amount of ethanol, preparing a 300 mu mol/L solution from a methyl jasmonate stock solution, placing the solution in a refrigerator for cooling to ensure that the temperature of the methyl jasmonate is reduced to 4-10 ℃, spraying the solution on the surface of 23d of the momordica grosvenori after pollination until the surface drips, and spraying the solution once in the morning, in the middle and at night and continuously for 5 d. And simultaneously, injecting methyl jasmonate into the main stem of the Momordica grosvenori plant in an injection mode, wherein the injection amount is 5-16 ml, and the injection speed is 4-8 ml/h.

Comparative example 4 was also set

2) the momordica grosvenori plants are not sprayed and injected with methyl jasmonate; other conditions were the same as in example 4.

(3) qRT-PCR detects the expression of UGT72B21 gene.

Collecting fruit, picking pulp, cutting into 2-4mm pieces, wrapping with tinfoil paper, quickly freezing in liquid nitrogen, and storing at-80 deg.C.

An ABI7500 real-time fluorescence quantitative PCR instrument is adopted.

The qRT-PCR detection result shows that the expression level of the momordica grosvenori UGT72B21 gene is 1 in the comparative example 4, while the expression level of the momordica grosvenori UGT72B21 gene is up to 20.33 in the example 4, which is improved by 1933% compared with the comparative example 4, and therefore, the example 4 can obviously promote the high expression of the momordica grosvenori UGT72B21 gene; HPLC detection results show that in comparative example 4, the mogroside V content is 0.80%, while in example 4, the mogroside V content is as high as 3.07%, which is improved by 283.75% compared with comparative example 4, and thus, example 4 can also significantly promote the improvement of mogroside V yield.

The number of modules and the processing scale described herein are intended to simplify the description of the invention. Applications, modifications and variations of the method of promoting expression of luo han guo UGT72B21 gene of the present invention will be apparent to those skilled in the art.

While embodiments of the invention have been described above, it is not limited to the applications set forth in the description and the embodiments, which are fully applicable to various fields of endeavor for which the invention may be embodied with additional modifications as would be readily apparent to those skilled in the art, and the invention is therefore not limited to the details given herein and to the embodiments shown and described without departing from the generic concept as defined by the claims and their equivalents.

SEQUENCE LISTING

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Claims

1. A method for promoting expression of Momordica grosvenori UGT72B21 gene is characterized by comprising the following steps: applying methyl jasmonate with the concentration of 50-400 mu mol/L in the tissue culture process and the cultivation process of the momordica grosvenori so as to promote the expression of the momordica grosvenori UGT72B21 gene;

in the tissue culture process of the momordica grosvenori, the methyl jasmonate is added into a solid culture medium in the tissue culture process of the momordica grosvenori, the solid culture medium in the tissue culture process of the momordica grosvenori consists of MS, 1.5mg/L6-BA, 0.3mg/L IBA, 3.5g/L agar, 30g/L sucrose and 1.0g/L active carbon, and the culture conditions in the tissue culture process of the momordica grosvenori are as follows: culturing for 30 days under the conditions of relative humidity of 60-66%, illumination intensity of 1400lux, illumination time of 8h/d and temperature of 21-25 ℃;

in the cultivation process of the momordica grosvenori, after 20-30 days after pollination of the momordica grosvenori, spraying the methyl jasmonate with the concentration of 50-400 mu mol/L to the surface of the momordica grosvenori until water drops on the surface of the momordica grosvenori, spraying the methyl jasmonate to the surface of the momordica grosvenori once in the morning, at the noon and at the evening, and continuously spraying the methyl jasmonate for 5 days, wherein in the cultivation process of the momordica grosvenori, the temperature of the methyl jasmonate sprayed to the surface of the momordica grosvenori is 4-10 ℃, meanwhile, the methyl jasmonate is injected into a main stem of a momordica grosvenori plant in an injection mode, the injection amount is 5-16 ml, and the injection rate is 4-8 ml/h;

also comprises the following steps: qRT-PCR detection of UGT72B21 Gene expression:

3) reverse transcribing the RNA to cDNA;

2. The method of promoting luo han guo UGT72B21 gene expression of claim 1, further comprising: preparing a jasmonic acid mother solution: dissolving methyl jasmonate in 2% (v/v) ethanol water solution to prepare 10000 mu mol/L mother liquor, and sterilizing with a 0.22 mu m microporous filter membrane to obtain sterilized methyl jasmonate mother liquor;