CN107258541A - Promote the method for Momordica grosvenori UGT7 gene expressions - Google Patents
Promote the method for Momordica grosvenori UGT7 gene expressions Download PDFInfo
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- CN107258541A CN107258541A CN201710629323.0A CN201710629323A CN107258541A CN 107258541 A CN107258541 A CN 107258541A CN 201710629323 A CN201710629323 A CN 201710629323A CN 107258541 A CN107258541 A CN 107258541A
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- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01H—NEW PLANTS OR NON-TRANSGENIC PROCESSES FOR OBTAINING THEM; PLANT REPRODUCTION BY TISSUE CULTURE TECHNIQUES
- A01H4/00—Plant reproduction by tissue culture techniques ; Tissue culture techniques therefor
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- A—HUMAN NECESSITIES
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- A01H4/00—Plant reproduction by tissue culture techniques ; Tissue culture techniques therefor
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Abstract
The invention provides a kind of method of promotion Momordica grosvenori UGT7 gene expressions, comprise the following steps：Step 1: Luohanguo With Plantlets of Tissue Culture is placed in into methyl jasmonate concentration to be cultivated in 50~400 μm of ol/L culture medium, Momordica grosvenori seedling is obtained；Step 2: during the Momordica grosvenori seedling obtained in cultivation step one, applying the ethanol solution that methyl jasmonate concentration is 50~400 μm of ol/L.The present invention by Luohanguo With Plantlets of Tissue Culture is placed in the culture medium containing methyl jasmonate cultivate and cultivate Momordica grosvenori during apply methyl jasmonate, the high of key gene UGT7 is expressed in rapid induction momordica glycoside V biosynthesis pathway in a short time, so as to promote the biosynthesis of momordica glycoside V, the accumulation for raising momordica glycoside V content is taken a firm foundation.
Description
Technical field
The present invention relates to plant biotechnology field, more specifically to a kind of promotion Momordica grosvenori UGT7 gene expressions
Method.
Background technology
Momordica glycoside V belongs to cucurbitane type tetracyclic triterpenes material, and this seminar early stage is according to Momordica grosvenori transcript profile number
According to having derived the possible biosynthesis pathways of sweet tea glycosides V.The precursor substance of momordica glycoside V biosynthesis is the phosphorus of iso-amylene two
Base pyrophosphoric acid (DMAPP) in sour (IPP) and 3,3- dimethyl alkene, the two is by mevalonic acid (MVA) and methylerythritol
Two approach of phosphorylation (MEP) are formed, and MVA approach occurs in kytoplasm, and MEP approach occurs in plastid.From above-mentioned two
The IPP or DMAPP of approach form Mang ox base pyrophosphoric acid (GPP), IPP and GPP through Mang ox base pyrophosphate synthase (GPS) catalysis
Under farnesyl pyrophosphate synthase (FPS) catalytic action so formed farnesyl pyrophosphate (FPP), then through squalene synthase
(SS), the catalysis of squalene epoxidase (SE) forms 2,3- oxidosqualenes, and cucurbit dienol synthase (CS) is further catalyzed
Cucurbit dienol is formed, finally in the presence of CYP450 enzymes and glucosyltransferase (UGT), momordica glycoside V is formed.
The generation of isoprenoid material is considered as on its biosynthesis way always by the strict regulation and control of speed limit enzymatic activity
Important regulating and controlling effect is played in footpath.As the rate-limiting enzyme in isoprene approach, the expression of UGT genes is to momordica glycoside V
Biosynthesis play decisive role.UGT genes are overexpressed, the accumulation of momordica glycoside V can be promoted；If on the contrary, suppression
The expression of UGT genes processed, the yield of momordica glycoside V will be significantly reduced.However, UGT genes are a supergene family, this problem
Group early-stage Study finds that UGT7 may participate in the route of synthesis of momordica glycoside V.Promotion Momordica grosvenori is there are no in the prior art
The report of UGT7 gene expressions.
The content of the invention
It is an object of the invention to solve at least the above, and provide the advantage that at least will be described later.
It is a still further object of the present invention to provide a kind of expression of promotion Momordica grosvenori UGT7 genes, it is by containing jasmine
The medium culture of jasmine acid methyl esters, and the ethanol solution containing methyl jasmonate is sprayed in the course of cultivation, Momordica grosvenori can be promoted
The expression of middle UGT7 genes, so as to promote the biosynthesis of momordica glycoside V, improves its yield.
Momordica grosvenori UGT7 gene tables are promoted there is provided one kind according to object of the present invention and further advantage in order to realize
The method reached, comprises the following steps：
Step 1: Luohanguo With Plantlets of Tissue Culture is placed in methyl jasmonate concentration to be cultivated in 50~400 μm of ol/L culture medium,
Obtain Momordica grosvenori seedling；
Step 2: in cultivation step one during obtained Momordica grosvenori seedling, apply methyl jasmonate concentration be 50~
400 μm of ol/L ethanol solution.
Preferably, culture medium also includes in the method for described promotion Momordica grosvenori UGT7 gene expressions, step one：MS、1
~2mg/L 6-BA, 0.2~0.5mg/L IBA, 3~4g/L agar, 20~40g/L sucrose, 0.5~2g/L activity
Charcoal.
Preferably, the training of Luohanguo With Plantlets of Tissue Culture in the method for described promotion Momordica grosvenori UGT7 gene expressions, step one
Foster method is：It is 60~66% in relative humidity, intensity of illumination is 1200~1500lux, and light application time is 8h/d, and temperature is 23
30d is cultivated under the conditions of ± 2 DEG C.
Preferably, in Momordica grosvenori cultivation in the method for described promotion Momordica grosvenori UGT7 gene expressions, step 2
In, after Pollination of Luohanguo after 20~30d, three methyl jasmonate concentration of sprinkling are molten for 50~400 μm of ol/L ethanol daily
Liquid, every plant is sprayed 200~300g every time, continuously sprays 5d.
Preferably, put in the method for described promotion Momordica grosvenori UGT7 gene expressions, step one in Luohanguo With Plantlets of Tissue Culture
Also include Luohanguo With Plantlets of Tissue Culture being placed in nutrient solution before cultivating in culture medium soaking 10 at temperature is 35 DEG C~45 DEG C
~20h；Wherein nutrient solution is composed of the following raw materials by weight：1~2 part of plant extraction liquid, 2~5 parts of gibberellin, apple butter 1~5
Part, 1~3 part of heteroauxin, 2~5 parts of sucrose, 0.1~0.5 part of methyl α-naphthyl acetate, 1~2 part of fulvic acid potassium, compound sodium nitrophenolate 0.1~
0.5 part, 20~30 parts of water；The plant extraction liquid is made up of following methods：By parts by weight for 2~5 parts cordate houttuynia, 1~2 part
Lindley Butterflybush Herb, 2~5 parts of Buddha's hand, 1~2 part of the bark of official magnolia, grind and be with the volume fraction that parts by weight are 20~30 parts successively
80~90%, 70~80% ethanol water is 60~90 DEG C of 12~24h of extraction in temperature, merges extract solution twice and produces plant
Thing extract solution.
Preferably, in cultivation Momordica grosvenori process in the method for described promotion Momordica grosvenori UGT7 gene expressions, step 2
In, 3~5d after Pollination of Luohanguo, every 2d apply one time of nutrition liquid, apply 3~5 times, every plant every time sprinkling 500~
1000g；The preparation method of wherein nutrient solution is：By parts by weight for 0.1~0.5 part CPPU, 0.1~0.2 part of brassin,
0.1~0.5 part of compound sodium nitrophenolate is dissolved in the absolute ethyl alcohol that parts by weight are 5~10 parts, is stirred, and then dissolving adds weight
Part is 5~10 parts of dimethyl sulfoxide (DMSO), 1~2 part of 2,4- dichlorophenoxyacetic acids, 5~10 parts of fulvic acid potassium, 10~20
The potassium sulfate, 20~50 parts of water of part stir and produced.
Preferably, put in the method for described promotion Momordica grosvenori UGT7 gene expressions, step one in Luohanguo With Plantlets of Tissue Culture
Water-loss reducer is additionally added when being cultivated in culture medium, the mass ratio of the water-loss reducer and culture medium is 5:0.1~0.5；The water conservation
Agent is made up of the raw material of following parts by weight：5~10 parts of sodium carboxymethylcellulose, 1~2 part of polyacrylic acid potassium, polyvinyl alcohol 1~2
Part, 0.1~0.5 part of zeolite powder, 0.1~0.2 part of walnut shell powder, 0.1~0.5 part of oyster shell powder, 1~2 part of bentonite, coconut shell flour
0.1~0.2 part, 0.1~0.5 part of shaddock peel powder, 1~2 part of alum, 0.5~1 part of alum, 1~2 part of urea, phosphoric acid
0.1~0.5 part of potassium dihydrogen, 5~10 parts of water, 0.1~0.5 part of acrylic acid.
Preferably, in the method for described promotion Momordica grosvenori UGT7 gene expressions, step 2 by Luohanguo With Plantlets of Tissue Culture extremely
When being cultivated in culture medium, while spray temperature is 40~50 DEG C of bactericidal liquid；The bactericidal liquid is by following raw material group
Into：0.1~0.5 part of Osthole, 0.1~0.5 part of aesculetin, 0.05~0.1 part of Wuyiencin, kasugarnycin 0.05~
0.1 part, 0.01~0.05 part of jinggangmeisu, 20~30 parts of pyrolkigneous liquid, 0.1~0.5 part of polyhexamethylene guanide, 50~100 parts of water,
0.01~0.05 part of berberine, 0.1~0.5 part of sodium benzoate, 1~2 part of methanesulfonic acid, 0.1~0.5 part of octyl trichlamide, lysozyme 1~
2 parts, 2~3 parts of lywallzyme, 0.1~0.5 part of astragalus polyose, 1~2 part of plant extraction liquid；Wherein, the preparation side of plant extraction liquid
Method is：It is 5~10 parts of Chinese scholartree root, 8~10 parts of Japanese Stephania Root, 5~10 parts of elecampane by parts by weight, is 70% with mass fraction
Ethanol solution extract twice, obtain medicinal extract, add the honey that parts by weight are 1~5 part, convert the water that parts by weight are 50~100 parts,
Produce plant extraction liquid
The present invention at least includes following beneficial effect：
1st, the present invention by Luohanguo With Plantlets of Tissue Culture by being placed in culture and cultivation sieve in the culture medium containing methyl jasmonate
Apply methyl jasmonate during Chinese fruit, in a short time key gene in rapid induction momordica glycoside V biosynthesis pathway
UGT7 high expression, so as to promote the biosynthesis of momordica glycoside V, heavily fortified point is laid in the accumulation for raising momordica glycoside V content
Real basis.
2nd, the present invention also applies CPPU, brassin, the compound nitrophenol in nutrient solution, nutrient solution during cultivation Momordica grosvenori
Sodium can promote Momordica grosvenori cell division, promote cell expansion elongation, promote fruit mast, improve momordica glycoside V content, together
When prepare nutrient solution when CPPU, brassin, compound sodium nitrophenolate are first dissolved in absolute ethyl alcohol, be conducive to these materials fully molten
Solution, to ensure that Momordica grosvenori can fully absorb these materials, promotes Momordica grosvenori growth.
3rd, the present invention also includes Luohanguo With Plantlets of Tissue Culture being placed in training before Luohanguo With Plantlets of Tissue Culture is placed in and cultivated in culture medium
Nutrient solution is soaked, gibberellin, heteroauxin, methyl α-naphthyl acetate, the compound sodium nitrophenolate in nutrient solution can promote Luohanguo With Plantlets of Tissue Culture cell division,
Promote cellular plasm flowing, improve cell viability, accelerate tissue-cultured seedling to grow；Contain plant extract in nutrient solution simultaneously
On the one hand the extract of cordate houttuynia, Lindley Butterflybush Herb, Buddha's hand, the bark of official magnolia in liquid, extract solution can provide nutrients for tissue-cultured seedling growth
Matter, on the other hand prevents tissue-cultured seedling from preventing the injury of pest and disease damage and bacterium from influenceing sweet tea mandarin orange V in follow-up Momordica grosvenori in incubation
Content improve.
4th, the present invention is additionally added water-loss reducer in the medium, and because culture medium desiccation is very fast, culture periphery is easily accumulated
Metabolite, causes the nutrition distribution pattern of culture medium uneven, is also unfavorable for culture and absorbs active ingredient.The guarantor of the present invention
Containing walnut shell powder, zeolite powder, oyster shell powder, coconut shell flour, shaddock peel powder there is microcellular structure not coagulated in culture medium in aqua
Gu when, the active ingredient in culture medium can be absorbed, the active ingredient in many culture mediums can be also adsorbed, so can effectively be kept
Nutritional ingredient, making the distribution of nutritional ingredient can slowly release than more uniform, adsorbed nutritional ingredient, be conducive to culture
Active ingredient is absorbed longer, so as to promote the growth of Luohanguo With Plantlets of Tissue Culture, is conducive to improving the content of momordica glycoside V.
5th, when Momordica grosvenori is placed in and cultivated in culture medium while spray bactericidal liquid, Osthole, bark of ash second in bactericidal liquid
Element, Wuyiencin, kasugarnycin, jinggangmeisu, lysozyme, lywallzyme can inactivate virus, fungi after being combined with virus；Its
Middle pyrolkigneous liquid has sterilization, promotes the effect such as plant growth；Plant extraction liquid in bactericidal liquid can be prevented in culture environment
Bacterium, virus and fungi influence the growth of Luohanguo With Plantlets of Tissue Culture, improve the content of sweet tea glycosides V in Lo Han Guo fruit, can also reduce pair
The requirement of culture environment.
Further advantage, target and the feature of the present invention embodies part by following explanation, and part will also be by this
The research and practice of invention and be understood by the person skilled in the art.
Embodiment
With reference to embodiment, the present invention is described in further detail, to make those skilled in the art with reference to specification
Word can be implemented according to this.
It should be noted that experimental method described in following embodiments, is conventional method unless otherwise specified, institute
Reagent and material are stated, unless otherwise specified, is commercially obtained.
Embodiment 1
A kind of method of promotion Momordica grosvenori UGT7 gene expressions, comprises the following steps：
Step 1: Luohanguo With Plantlets of Tissue Culture is placed in into methyl jasmonate concentration to be cultivated in 50 μm of ol/L culture medium, obtain
Momordica grosvenori seedling；
Step 2: during the Momordica grosvenori seedling obtained in cultivation step one, it is 50 μ to apply methyl jasmonate concentration
Mol/L ethanol solution.
Culture medium also includes in the step one：MS, 1mg/L 6-BA, 0.2mg/L IBA, 3g/L agar, 20g/L
Sucrose, 0.5g/L activated carbon；Wherein, 6-BA is 6- benzyl aminoadenines, and IBA is indolebutyric acid.
The cultural method of Luohanguo With Plantlets of Tissue Culture is in the step one：It is 60% in relative humidity, intensity of illumination is
1200lux, light application time is 8h/d, and temperature is to cultivate 30d under the conditions of 21 DEG C.
In the step 2 in Momordica grosvenori cultivation, after Pollination of Luohanguo after 20d, three jasmonics are sprayed daily
Methyl acetate concentrations are 50 μm of ol/L ethanol solution, and every plant is sprayed 200g every time, continuously sprays 5d.
Embodiment 2
A kind of method of promotion Momordica grosvenori UGT7 gene expressions, comprises the following steps：
Step 1: Luohanguo With Plantlets of Tissue Culture is placed in into methyl jasmonate concentration to be cultivated in 250 μm of ol/L culture medium, obtain
Momordica grosvenori seedling；
Step 2: during the Momordica grosvenori seedling obtained in cultivation step one, it is 200 μ to apply methyl jasmonate concentration
Mol/L ethanol solution.
Culture medium also includes in the step one：MS, 1.5mg/L 6-BA, 0.3mg/L IBA, 3.5g/L agar,
30g/L sucrose, 1g/L activated carbon；Wherein, 6-BA is 6- benzyl aminoadenines, and IBA is indolebutyric acid.
The cultural method of Luohanguo With Plantlets of Tissue Culture is in the step one：It is 63% in relative humidity, intensity of illumination is
1400lux, light application time is 8h/d, and temperature is to cultivate 30d under the conditions of 23 DEG C.
In the step 2 in Momordica grosvenori cultivation, after Pollination of Luohanguo after 25d, three jasmonics are sprayed daily
Methyl acetate concentrations are 200 μm of ol/L ethanol solution, and every plant is sprayed 250g every time, continuously sprays 5d.
Embodiment 3
A kind of method of promotion Momordica grosvenori UGT7 gene expressions, comprises the following steps：
Step 1: Luohanguo With Plantlets of Tissue Culture is placed in into methyl jasmonate concentration to be cultivated in 400 μm of ol/L culture medium, obtain
Momordica grosvenori seedling；
Step 2: during the Momordica grosvenori seedling obtained in cultivation step one, it is 400 μ to apply methyl jasmonate concentration
Mol/L ethanol solution.
Culture medium also includes in the step one：MS, 2mg/L 6-BA, 0.5mg/L IBA, 4g/L agar, 40g/L
Sucrose, 2g/L activated carbon；Wherein, 6-BA is 6- benzyl aminoadenines, and IBA is indolebutyric acid.
The cultural method of Luohanguo With Plantlets of Tissue Culture is in the step one：It is 66% in relative humidity, intensity of illumination is
1500lux, light application time is 8h/d, and temperature is to cultivate 30d under the conditions of 25 DEG C.
In the step 2 in Momordica grosvenori cultivation, after Pollination of Luohanguo after 30d, three jasmonics are sprayed daily
Methyl acetate concentrations are 400 μm of ol/L ethanol solution, and every plant is sprayed 300g every time, continuously sprays 5d.
Embodiment 4
A kind of method of promotion Momordica grosvenori UGT7 gene expressions, comprises the following steps：
Step 1: Luohanguo With Plantlets of Tissue Culture is placed in into methyl jasmonate concentration to be cultivated in 50 μm of ol/L culture medium, obtain
Momordica grosvenori seedling；
Step 2: during the Momordica grosvenori seedling obtained in cultivation step one, it is 50 μ to apply methyl jasmonate concentration
Mol/L ethanol solution.
Culture medium also includes in the step one：MS, 1mg/L 6-BA, 0.2mg/L IBA, 3g/L agar, 20g/L
Sucrose, 0.5g/L activated carbon；Wherein, 6-BA is 6- benzyl aminoadenines, and IBA is indolebutyric acid.
The cultural method of Luohanguo With Plantlets of Tissue Culture is in the step one：It is 60% in relative humidity, intensity of illumination is
1200lux, light application time is 8h/d, and temperature is to cultivate 30d under the conditions of 21 DEG C.
In the step 2 in Momordica grosvenori cultivation, after Pollination of Luohanguo after 20d, three jasmonics are sprayed daily
Methyl acetate concentrations are 50 μm of ol/L ethanol solution, and every plant is sprayed 200g every time, continuously sprays 5d.
Also including Luohanguo With Plantlets of Tissue Culture is put before Luohanguo With Plantlets of Tissue Culture is placed in and cultivated in culture medium in the step one
In nutrient solution 10h is soaked at temperature is 35 DEG C；Wherein nutrient solution is composed of the following raw materials by weight：1 part of plant extraction liquid,
2 parts of gibberellin, 1 part of apple butter, 1 part of heteroauxin, 2 parts of sucrose, 0.1 part of methyl α-naphthyl acetate, 1 part of fulvic acid potassium, compound sodium nitrophenolate 0.1
Part, 20 parts of water；The plant extraction liquid is made up of following methods：By parts by weight for 2 parts cordate houttuynia, 1 part of Lindley Butterflybush Herb, 2 parts
Buddha's hand, 1 part of the bark of official magnolia, grind the ethanol water for being successively 80%, 70% with the volume fraction that parts by weight are 20 parts
It is 60 DEG C of extraction 12h in temperature, merges extract solution twice and produce plant extraction liquid.
In the step 2 during cultivation Momordica grosvenori, the 3d after Pollination of Luohanguo applies one time of nutrition liquid every 2d,
Apply 3 times, every plant is sprayed 500g every time；The preparation method of wherein nutrient solution is：By parts by weight for 0.1 part CPPU, 0.1 part
Brassin, 0.1 part of compound sodium nitrophenolate be dissolved in during parts by weight are 5 parts of absolute ethyl alcohol, stir, then dissolving adds weight
Part for 5 parts dimethyl sulfoxide (DMSO), 1 part of 2,4- dichlorophenoxyacetic acids, 5 parts of fulvic acid potassium, 10 parts of potassium sulfate, 20 parts
Water stir and produce.
Be additionally added water-loss reducer when Luohanguo With Plantlets of Tissue Culture is placed in and cultivated in culture medium in the step one, the water-loss reducer with
The mass ratio of culture medium is 5:0.1；The water-loss reducer is made up of the raw material of following parts by weight：5 parts of sodium carboxymethylcellulose, poly- third
1 part of olefin(e) acid potassium, 1 part of polyvinyl alcohol, 0.1 part of zeolite powder, 0.1 part of walnut shell powder, 0.1 part of oyster shell powder, 1 part of bentonite, coconut husk
0.1 part of powder, 0.1 part of shaddock peel powder, 1 part of alum, 0.5 part of alum, 1 part of urea, 0.1 part of potassium dihydrogen phosphate, water 5
Part, 0.1 part of acrylic acid.
In the step 2 by Luohanguo With Plantlets of Tissue Culture as being cultivated in culture medium when, while spray temperature is 40 DEG C of sterilization
Liquid；The bactericidal liquid is composed of the following raw materials by weight：0.1 part of Osthole, 0.1 part of aesculetin, 0.05 part of Wuyiencin,
0.05 part of kasugarnycin, 0.01 part of jinggangmeisu, 20 parts of pyrolkigneous liquid, 0.1 part of polyhexamethylene guanide, 50 parts of water, berberine 0.01
Part, 0.1 part of sodium benzoate, 1 part of methanesulfonic acid, 0.1 part of octyl trichlamide, 1 part of lysozyme, 2 parts of lywallzyme, 0.1 part of astragalus polyose, plant
1 part of extract solution；Wherein, the preparation method of plant extraction liquid is：By the Chinese scholartree root, 8 parts of Japanese Stephania Root, 5 parts of soil that parts by weight are 5 parts
The banksia rose, is extracted twice for 70% ethanol solution with mass fraction, obtains medicinal extract, is added the honey that parts by weight are 1 part, is converted weight
Part is 50 parts of water, produces plant extraction liquid.
Embodiment 5
A kind of method of promotion Momordica grosvenori UGT7 gene expressions, comprises the following steps：
Step 1: Luohanguo With Plantlets of Tissue Culture is placed in into methyl jasmonate concentration to be cultivated in 250 μm of ol/L culture medium, obtain
Momordica grosvenori seedling；
Step 2: during the Momordica grosvenori seedling obtained in cultivation step one, it is 200 μ to apply methyl jasmonate concentration
Mol/L ethanol solution.
Culture medium also includes in the step one：MS, 1.5mg/L 6-BA, 0.3mg/L IBA, 3.5g/L agar,
30g/L sucrose, 1g/L activated carbon；Wherein, 6-BA is 6- benzyl aminoadenines, and IBA is indolebutyric acid.
The cultural method of Luohanguo With Plantlets of Tissue Culture is in the step one：It is 63% in relative humidity, intensity of illumination is
1400lux, light application time is 8h/d, and temperature is to cultivate 30d under the conditions of 23 DEG C.
In the step 2 in Momordica grosvenori cultivation, after Pollination of Luohanguo after 25d, three jasmonics are sprayed daily
Methyl acetate concentrations are 200 μm of ol/L ethanol solution, and every plant is sprayed 250g every time, continuously sprays 5d.
Also including Luohanguo With Plantlets of Tissue Culture is put before Luohanguo With Plantlets of Tissue Culture is placed in and cultivated in culture medium in the step one
In nutrient solution 15h is soaked at temperature is 40 DEG C；Wherein nutrient solution is composed of the following raw materials by weight：2 parts of plant extraction liquid,
4 parts of gibberellin, 3 parts of apple butter, 2 parts of heteroauxin, 3 parts of sucrose, 0.3 part of methyl α-naphthyl acetate, 2 parts of fulvic acid potassium, compound sodium nitrophenolate 0.3
Part, 25 parts of water；The plant extraction liquid is made up of following methods：By parts by weight for 3 parts cordate houttuynia, 2 parts of Lindley Butterflybush Herb, 3 parts
Buddha's hand, 1 part of the bark of official magnolia, grind the ethanol water for being successively 85%, 75% with the volume fraction that parts by weight are 30 parts
It is 80 DEG C of extraction 18h in temperature, merges extract solution twice and produce plant extraction liquid.
In the step 2 during cultivation Momordica grosvenori, the 4d after Pollination of Luohanguo applies one time of nutrition liquid every 2d,
Apply 4 times, every plant is sprayed 700g every time；The preparation method of wherein nutrient solution is：By parts by weight for 0.3 part CPPU, 0.1 part
Brassin, 0.3 part of compound sodium nitrophenolate be dissolved in during parts by weight are 7 parts of absolute ethyl alcohol, stir, then dissolving adds weight
Part for 7 parts dimethyl sulfoxide (DMSO), 1 part of 2,4- dichlorophenoxyacetic acids, 8 parts of fulvic acid potassium, 15 parts of potassium sulfate, 35 parts
Water stir and produce.
Be additionally added water-loss reducer when Luohanguo With Plantlets of Tissue Culture is placed in and cultivated in culture medium in the step one, the water-loss reducer with
The mass ratio of culture medium is 5:0.3；The water-loss reducer is made up of the raw material of following parts by weight：7 parts of sodium carboxymethylcellulose, poly- third
2 parts of olefin(e) acid potassium, 1 part of polyvinyl alcohol, 0.3 part of zeolite powder, 0.1 part of walnut shell powder, 0.3 part of oyster shell powder, 1 part of bentonite, coconut husk
0.1 part of powder, 0.3 part of shaddock peel powder, 1 part of alum, 0.8 part of alum, 1 part of urea, 0.3 part of potassium dihydrogen phosphate, water 8
Part, 0.3 part of acrylic acid.
In the step 2 by Luohanguo With Plantlets of Tissue Culture as being cultivated in culture medium when, while spray temperature is 45 DEG C of sterilization
Liquid；The bactericidal liquid is composed of the following raw materials by weight：0.3 part of Osthole, 0.3 part of aesculetin, 0.08 part of Wuyiencin,
0.07 part of kasugarnycin, 0.03 part of jinggangmeisu, 25 parts of pyrolkigneous liquid, 0.3 part of polyhexamethylene guanide, 70 parts of water, berberine 0.03
Part, 0.3 part of sodium benzoate, 2 parts of methanesulfonic acid, 0.3 part of octyl trichlamide, 1 part of lysozyme, 2 parts of lywallzyme, 0.3 part of astragalus polyose, plant
1 part of extract solution；Wherein, the preparation method of plant extraction liquid is：By the Chinese scholartree root, 9 parts of Japanese Stephania Root, 7 parts of soil that parts by weight are 8 parts
The banksia rose, is extracted twice for 70% ethanol solution with mass fraction, obtains medicinal extract, is added the honey that parts by weight are 3 parts, is converted weight
Part is 70 parts of water, produces plant extraction liquid.
Embodiment 6
A kind of method of promotion Momordica grosvenori UGT7 gene expressions, comprises the following steps：
Step 1: Luohanguo With Plantlets of Tissue Culture is placed in into methyl jasmonate concentration to be cultivated in 400 μm of ol/L culture medium, obtain
Momordica grosvenori seedling；
Step 2: during the Momordica grosvenori seedling obtained in cultivation step one, it is 400 μ to apply methyl jasmonate concentration
Mol/L ethanol solution.
Culture medium also includes in the step one：MS, 2mg/L 6-BA, 0.5mg/L IBA, 4g/L agar, 40g/L
Sucrose, 2g/L activated carbon；Wherein, 6-BA is 6- benzyl aminoadenines, and IBA is indolebutyric acid.
The cultural method of Luohanguo With Plantlets of Tissue Culture is in the step one：It is 66% in relative humidity, intensity of illumination is
1500lux, light application time is 8h/d, and temperature is to cultivate 30d under the conditions of 25 DEG C.
In the step 2 in Momordica grosvenori cultivation, after Pollination of Luohanguo after 30d, three jasmonics are sprayed daily
Methyl acetate concentrations are 400 μm of ol/L ethanol solution, and every plant is sprayed 300g every time, continuously sprays 5d.
Also including Luohanguo With Plantlets of Tissue Culture is placed in before Luohanguo With Plantlets of Tissue Culture is placed in and cultivated in culture medium in described rapid one
In nutrient solution 20h is soaked at temperature is 45 DEG C；Wherein nutrient solution is composed of the following raw materials by weight：It is 2 parts of plant extraction liquid, red
5 parts of mycin, 5 parts of apple butter, 3 parts of heteroauxin, 5 parts of sucrose, 0.5 part of methyl α-naphthyl acetate, 2 parts of fulvic acid potassium, compound sodium nitrophenolate 0.5
Part, 30 parts of water；The plant extraction liquid is made up of following methods：By parts by weight for 5 parts cordate houttuynia, 2 parts of Lindley Butterflybush Herb, 5 parts
Buddha's hand, 2 parts of the bark of official magnolia, grind the ethanol water for being successively 90%, 80% with the volume fraction that parts by weight are 30 parts
It is 90 DEG C of extraction 24h in temperature, merges extract solution twice and produce plant extraction liquid.
In the step 2 during cultivation Momordica grosvenori, the 5d after Pollination of Luohanguo applies one time of nutrition liquid every 2d,
Apply 5 times, every plant is sprayed 1000g every time；The preparation method of wherein nutrient solution is：By parts by weight for 0.5 part CPPU, 0.2
The brassin, 0.5 part of compound sodium nitrophenolate of part are dissolved in the absolute ethyl alcohol that parts by weight are 10 parts, are stirred, and then dissolving adds weight
It is 10 parts of dimethyl sulfoxide (DMSO), 2 parts of 2,4- dichlorophenoxyacetic acids, 10 parts of fulvic acid potassium, 120 parts of sulfuric acid to measure part
Potassium, 50 parts of water stir and produced.
Be additionally added water-loss reducer when Luohanguo With Plantlets of Tissue Culture is placed in and cultivated in culture medium in the step one, the water-loss reducer with
The mass ratio of culture medium is 5:0.5；The water-loss reducer is made up of the raw material of following parts by weight：It is 10 parts of sodium carboxymethylcellulose, poly-
2 parts of potassium acrylate, 2 parts of polyvinyl alcohol, 0.5 part of zeolite powder, 0.2 part of walnut shell powder, 0.5 part of oyster shell powder, 2 parts of bentonite, coconut palm
0.2 part of shell powder, 0.5 part of shaddock peel powder, 2 parts of alum, 1 part of alum, 2 parts of urea, 0.5 part of potassium dihydrogen phosphate, water 10
Part, 0.5 part of acrylic acid.
In the step 2 by Luohanguo With Plantlets of Tissue Culture as being cultivated in culture medium when, while spray temperature is 50 DEG C of sterilization
Liquid；The bactericidal liquid is composed of the following raw materials by weight：0.5 part of Osthole, 0.5 part of aesculetin, 0.1 part of Wuyiencin,
0.1 part of kasugarnycin, 0.05 part of jinggangmeisu, 30 parts of pyrolkigneous liquid, 00.5 part of polyhexamethylene guanide, 100 parts of water, berberine 0.05
Part, 0.5 part of sodium benzoate, 2 parts of methanesulfonic acid, 0.5 part of octyl trichlamide, 2 parts of lysozyme, 3 parts of lywallzyme, 0.5 part of astragalus polyose, plant
2 parts of extract solution；Wherein, the preparation method of plant extraction liquid is：By parts by weight for 10 parts Chinese scholartree root, 10 parts of Japanese Stephania Root, 10 parts
Elecampane, extracted twice for 70% ethanol solution with mass fraction, obtain medicinal extract, added the honey that parts by weight are 5 parts, convert
Parts by weight are 100 parts of water, produce plant extraction liquid.
Comparative example 1
A kind of Momordica grosvenori implantation methods, comprise the following steps：
Cultivated Step 1: Luohanguo With Plantlets of Tissue Culture is placed in culture medium, obtain Momordica grosvenori seedling；
Step 2: the Momordica grosvenori seedling obtained in cultivation step one, obtains Lo Han Guo fruit.
Culture medium includes in the step one：MS, 1mg/L 6-BA, 0.2mg/L IBA, 3g/L agar, 20g/L
The activated carbon of sucrose, 0.5g/L；Wherein, 6-BA is 6- benzyl aminoadenines, and IBA is indolebutyric acid.
The cultural method of Luohanguo With Plantlets of Tissue Culture is in the step one：It is 60% in relative humidity, intensity of illumination is
1200lux, light application time is 8h/d, and temperature is to cultivate 30d under the conditions of 21 DEG C.
Comparative example 2
A kind of Momordica grosvenori implantation methods, comprise the following steps：
Cultivated Step 1: Luohanguo With Plantlets of Tissue Culture is placed in culture medium, obtain Momordica grosvenori seedling；
Step 2: the Momordica grosvenori seedling obtained in cultivation step one, obtains Lo Han Guo fruit.
Culture medium includes in the step one：MS, 1mg/L 6-BA, 0.2mg/L IBA, 3g/L agar, 20g/L
The activated carbon of sucrose, 0.5g/L；Wherein, 6-BA is 6- benzyl aminoadenines, and IBA is indolebutyric acid.
The cultural method of Luohanguo With Plantlets of Tissue Culture is in the step one：It is 60% in relative humidity, intensity of illumination is
1200lux, light application time is 8h/d, and temperature is to cultivate 30d under the conditions of 21 DEG C.
In the step 2 during cultivation Momordica grosvenori, the 3d after Pollination of Luohanguo applies one time of nutrition liquid every 2d,
Apply 3 times, every plant is sprayed 500g every time；The preparation method of wherein nutrient solution is：By parts by weight for 0.1 part CPPU, 0.1 part
Brassin, 0.1 part of compound sodium nitrophenolate be dissolved in during parts by weight are 5 parts of absolute ethyl alcohol, stir, then dissolving adds weight
Part for 5 parts dimethyl sulfoxide (DMSO), 1 part of 2,4- dichlorophenoxyacetic acids, 5 parts of fulvic acid potassium, 10 parts of potassium sulfate, 20 parts
Water stir and produce.
Comparative example 3
A kind of Momordica grosvenori implantation methods, comprise the following steps：
Cultivated Step 1: Luohanguo With Plantlets of Tissue Culture is placed in culture medium, obtain Momordica grosvenori seedling；
Step 2: the Momordica grosvenori seedling obtained in cultivation step one, obtains Lo Han Guo fruit.
Culture medium includes in the step one：MS, 1mg/L 6-BA, 0.2mg/L IBA, 3g/L agar, 20g/L
The activated carbon of sucrose, 0.5g/L；Wherein, 6-BA is 6- benzyl aminoadenines, and IBA is indolebutyric acid.
The cultural method of Luohanguo With Plantlets of Tissue Culture is in the step one：It is 60% in relative humidity, intensity of illumination is
1200lux, light application time is 8h/d, and temperature is to cultivate 30d under the conditions of 21 DEG C.
Be additionally added water-loss reducer when Luohanguo With Plantlets of Tissue Culture is placed in and cultivated in culture medium in the step one, the water-loss reducer with
The mass ratio of culture medium is 5:0.1；The water-loss reducer is made up of the raw material of following parts by weight：5 parts of sodium carboxymethylcellulose, poly- third
1 part of olefin(e) acid potassium, 1 part of polyvinyl alcohol, 0.1 part of zeolite powder, 0.1 part of walnut shell powder, 0.1 part of oyster shell powder, 1 part of bentonite, coconut husk
0.1 part of powder, 0.1 part of shaddock peel powder, 1 part of alum, 0.5 part of alum, 1 part of urea, 0.1 part of potassium dihydrogen phosphate, water 5
Part, 0.1 part of acrylic acid.
Comparative example 4
A kind of Momordica grosvenori implantation methods, comprise the following steps：
Cultivated Step 1: Luohanguo With Plantlets of Tissue Culture is placed in culture medium, obtain Momordica grosvenori seedling；
Step 2: the Momordica grosvenori seedling obtained in cultivation step one, obtains Lo Han Guo fruit.
Culture medium includes in the step one：MS, 1mg/L 6-BA, 0.2mg/L IBA, 3g/L agar, 20g/L
The activated carbon of sucrose, 0.5g/L；Wherein, 6-BA is 6- benzyl aminoadenines, and IBA is indolebutyric acid.
The cultural method of Luohanguo With Plantlets of Tissue Culture is in the step one：It is 60% in relative humidity, intensity of illumination is
1200lux, light application time is 8h/d, and temperature is to cultivate 30d under the conditions of 21 DEG C.
Also including Luohanguo With Plantlets of Tissue Culture is put before Luohanguo With Plantlets of Tissue Culture is placed in and cultivated in culture medium in the step one
In nutrient solution 10h is soaked at temperature is 35 DEG C；Wherein nutrient solution is composed of the following raw materials by weight：1 part of plant extraction liquid,
2 parts of gibberellin, 1 part of apple butter, 1 part of heteroauxin, 2 parts of sucrose, 0.1 part of methyl α-naphthyl acetate, 1 part of fulvic acid potassium, compound sodium nitrophenolate 0.1
Part, 20 parts of water；The plant extraction liquid is made up of following methods：By parts by weight for 2 parts cordate houttuynia, 1 part of Lindley Butterflybush Herb, 2 parts
Buddha's hand, 1 part of the bark of official magnolia, grind the ethanol water for being successively 80%, 70% with the volume fraction that parts by weight are 20 parts
It is 60 DEG C of extraction 12h in temperature, merges extract solution twice and produce plant extraction liquid.
In the step 2 by Luohanguo With Plantlets of Tissue Culture as being cultivated in culture medium when, while spray temperature is 40 DEG C of sterilization
Liquid；The bactericidal liquid is composed of the following raw materials by weight：0.1 part of Osthole, 0.1 part of aesculetin, 0.05 part of Wuyiencin,
0.05 part of kasugarnycin, 0.01 part of jinggangmeisu, 20 parts of pyrolkigneous liquid, 0.1 part of polyhexamethylene guanide, 50 parts of water, berberine 0.01
Part, 0.1 part of sodium benzoate, 1 part of methanesulfonic acid, 0.1 part of octyl trichlamide, 1 part of lysozyme, 2 parts of lywallzyme, 0.1 part of astragalus polyose, plant
1 part of extract solution；Wherein, the preparation method of plant extraction liquid is：By the Chinese scholartree root, 8 parts of Japanese Stephania Root, 5 parts of soil that parts by weight are 5 parts
The banksia rose, is extracted twice for 70% ethanol solution with mass fraction, obtains medicinal extract, is added the honey that parts by weight are 1 part, is converted weight
Part is 50 parts of water, produces plant extraction liquid.
The Lo Han Guo fruit obtained using above-described embodiment 1~6, the method for comparative example 1~4, for UGT7 gene expression amounts
And sweet tea glycosides V content is measured, specific method and result are as follows.
1st, UGT7 gene expressions quantity measuring method：Using ABI7500 real-time fluorescence quantitative PCR instrument, detected using qRT-PCR
The expression of UGT7 genes, is comprised the following steps that：
Step 1: collection Lo Han Guo fruit, picking pulp is cut into 2-4mm fritters and wrapped respectively with masking foil, put immediately
It is quick-frozen in liquid nitrogen, be stored in -80 DEG C it is standby.
Step 2: first extracting Lo Han Guo fruit total serum IgE using improved Trizol method；
Step 3: the reaction system for being cDNA by RNA reverse transcriptions is μ L, the PrimeScript RT Enzyme of RNA 10.0
μ L, the RNase Free dH of 1.0 μ L, RT Primer Mix of Mix I, 1.0 μ L, 5 × PrimeScript Buffer 2 4.02O
4.0μL；Reaction condition is 37 DEG C of (15min) → 85 DEG C (5s) → 4 DEG C；
Step 4: using the software Design primers of Primer Premier 5.0, being had by raw work bioengineering (Shanghai) share
Limit company synthesizes, wherein, UGT7 primer sequences are：
FP：CTCCAGTATCGGTGGTCTTCT, RP：GCTTCAAACTTATCCTTCGGT；
Reference gene UBQ5, sequence is：
FP：ATAAAAGACCCAGCACCACATTC, RP：CCCTTGCCGACTACAACATCC；
Step 5: qRT-PCR reaction systems (20 μ L) are SYBR Premix Ex Taq II (Tli RNaseH Plus)
(2 ×) 10.0 μ L, PCR Forward Primer (10 μM) 0.8 μ L, PCR Reverse Primer (10 μM) 0.8 μ L, ROX
Reference Dye of Dye II (50 ×) 0.4 μ L, Template 2.0 μ L, dH2O 6.0μL.Reaction condition is 95 DEG C
(30s), then carries out 40 cycles [95 DEG C (5s), 95 DEG C (34s)].
2nd, the measure of momordica glycoside V content：Contained using momordica glycoside V in high effective liquid chromatography for measuring Momordica grosvenori
Amount.
The measurement of Lo Han Guo fruit UGT7 gene expression amounts and sweet tea glycosides V content that embodiment 1~6, comparative example 1~4 are obtained
As a result it is as shown in table 1 below, wherein experimental data be on 20 Momordica grosvenori fruit trees that each above-mentioned embodiment is obtained optionally
The Lo Han Guo fruit obtained in wherein 5 carries out what parallel test was obtained.
The Lo Han Guo fruit UGT7 gene expression amounts of embodiment 1~6 and sweet tea glycosides V content are above as can be seen from Table 1
Comparative example 1~4.Sieve is cultivated and cultivates it can therefore be concluded that Luohanguo With Plantlets of Tissue Culture is placed in the culture medium containing methyl jasmonate
Apply methyl jasmonate during Chinese fruit, gene UGT7 high expression can be promoted, so as to promote the biological conjunction of momordica glycoside V
Into raising momordica glycoside V content.Further the from comparative example 1 and the contrast of comparative example 2 it can be seen that in cultivation Momordica grosvenori process
In simultaneously apply nutrient solution can promote UGT7 gene expressions, so as to improve sweet tea glycosides V content；Can from comparative example 1 and the contrast of comparative example 3
To add water-loss reducer when finding out that Luohanguo With Plantlets of Tissue Culture is placed in and cultivated in culture medium, the raising of momordica glycoside V content can be promoted；
Contrasted from comparative example 1 and 4 it can be seen that soaking nutrient solution before Luohanguo With Plantlets of Tissue Culture is placed in medium culture and cultivating
Bactericidal liquid cocoa is sprayed in journey and promotes the growth of Momordica grosvenori, and then makes UGT7 high expression, so as to promote momordica glycoside V content
Raising.
Table 1 not be the same as Example Momordica grosvenori UGT7 gene expression amounts and sweet tea glycosides V content
| Embodiment | Sweet tea glycosides V content/(w/w%) | UGT7 gene expression amounts |
| Embodiment 1 | 2.09 | 12.7 |
| Embodiment 2 | 2.19 | 13.9 |
| Embodiment 3 | 2.32 | 15.3 |
| Embodiment 4 | 3.06 | 22.3 |
| Embodiment 5 | 3.13 | 23.8 |
| Embodiment 6 | 3.24 | 24.8 |
| Comparative example 1 | 0.72 | 3.6 |
| Comparative example 2 | 1.28 | 5.3 |
| Comparative example 3 | 1.35 | 6.5 |
| Comparative example 4 | 1.62 | 8.9 |
Although embodiment of the present invention is disclosed as above, it is not restricted in specification and embodiment listed
With it can be applied to various suitable the field of the invention completely, can be easily for those skilled in the art
Other modification is realized, therefore under the universal limited without departing substantially from claim and equivalency range, the present invention is not limited
In specific details and shown here as the embodiment with description.
Claims (8)
1. a kind of method of promotion Momordica grosvenori UGT7 gene expressions, it is characterised in that comprise the following steps：
Step 1: Luohanguo With Plantlets of Tissue Culture is placed in into methyl jasmonate concentration to be cultivated in 50~400 μm of ol/L culture medium, obtain
Momordica grosvenori seedling；
Step 2: during the Momordica grosvenori seedling obtained in cultivation step one, it is 50~400 μ to apply methyl jasmonate concentration
Mol/L ethanol solution.
2. promote the method for Momordica grosvenori UGT7 gene expressions as claimed in claim 1, it is characterised in that culture medium in step one
Also include：MS, 1~2mg/L 6-BA, 0.2~0.5mg/L IBA, 3~4g/L agar, 20~40g/L sucrose, 0.5
~2g/L activated carbon.
3. promote the method for Momordica grosvenori UGT7 gene expressions as claimed in claim 1, it is characterised in that Momordica grosvenori in step one
The cultural method of tissue-cultured seedling is：It is 60~66% in relative humidity, intensity of illumination is 1200~1500lux, and light application time is 8h/
D, temperature be 23 ± 2 DEG C under the conditions of cultivate 30d.
4. promote the method for Momordica grosvenori UGT7 gene expressions as claimed in claim 1, it is characterised in that in arhat in step 2
In fruit cultivation, after Pollination of Luohanguo after 20~30d, three methyl jasmonate concentration of sprinkling are 50~400 μm of ol/ daily
L ethanol solution, every plant is sprayed 200~300g every time, continuously sprays 5d.
5. promote the method for Momordica grosvenori UGT7 gene expressions as claimed in claim 1, it is characterised in that in arhat in step one
It is 35 DEG C~45 that fruit tissue-cultured seedling, which is placed in culture medium before culture also including Luohanguo With Plantlets of Tissue Culture is placed in nutrient solution in temperature,
10~20h is soaked at DEG C；Wherein nutrient solution is composed of the following raw materials by weight：1~2 part of plant extraction liquid, 2~5 parts of gibberellin,
1~5 part of apple butter, 1~3 part of heteroauxin, 2~5 parts of sucrose, 0.1~0.5 part of methyl α-naphthyl acetate, 1~2 part of fulvic acid potassium, multiple nitre
0.1~0.5 part of phenol sodium, 20~30 parts of water；The plant extraction liquid is made up of following methods：By the fish raw meat that parts by weight are 2~5 parts
Grass, 1~2 part of Lindley Butterflybush Herb, 2~5 parts of Buddha's hand, 1~2 part of the bark of official magnolia, it is successively 20~30 parts with parts by weight to grind
The ethanol water that volume fraction is 80~90%, 70~80% is 60~90 DEG C of 12~24h of extraction in temperature, and merging is carried twice
Liquid is taken to produce plant extraction liquid.
6. promote the method for Momordica grosvenori UGT7 gene expressions as claimed in claim 1, it is characterised in that in cultivation in step 2
During Momordica grosvenori, 3~5d after Pollination of Luohanguo applies one time of nutrition liquid every 2d, applies 3~5 times, every plant of sprinkling every time
500~1000g；The preparation method of wherein nutrient solution is：CPPU, 0.1~0.2 part of rue by parts by weight for 0.1~0.5 part
Tongue element, 0.1~0.5 part of compound sodium nitrophenolate are dissolved in the absolute ethyl alcohol that parts by weight are 5~10 parts, are stirred, and then dissolving adds
Parts by weight are 5~10 parts of dimethyl sulfoxide (DMSO), 1~2 part of 2,4- dichlorophenoxyacetic acids, 5~10 parts of fulvic acid potassium, 10
~20 parts of potassium sulfate, 20~50 parts of water stir and produced.
7. promote the method for Momordica grosvenori UGT7 gene expressions as claimed in claim 1, it is characterised in that in arhat in step one
Fruit tissue-cultured seedling is placed in and water-loss reducer is additionally added when being cultivated in culture medium, and the mass ratio of the water-loss reducer and culture medium is 5:0.1~
0.5；The water-loss reducer is made up of the raw material of following parts by weight：5~10 parts of sodium carboxymethylcellulose, 1~2 part of polyacrylic acid potassium,
1~2 part of polyvinyl alcohol, 0.1~0.5 part of zeolite powder, 0.1~0.2 part of walnut shell powder, 0.1~0.5 part of oyster shell powder, bentonite 1
~2 parts, 0.1~0.2 part of coconut shell flour, 0.1~0.5 part of shaddock peel powder, 1~2 part of alum, 0.5~1 part of alum, urine
1~2 part of element, 0.1~0.5 part of potassium dihydrogen phosphate, 5~10 parts of water, 0.1~0.5 part of acrylic acid.
8. promote the method for Momordica grosvenori UGT7 gene expressions as claimed in claim 1, it is characterised in that by arhat in step 2
When fruit tissue-cultured seedling in culture medium as cultivating, while spray temperature is 40~50 DEG C of bactericidal liquid；The bactericidal liquid is by following heavy
Measure part raw material composition：0.1~0.5 part of Osthole, 0.1~0.5 part of aesculetin, 0.05~0.1 part of Wuyiencin, spring thunder are mould
0.05~0.1 part of element, 0.01~0.05 part of jinggangmeisu, 20~30 parts of pyrolkigneous liquid, 0.1~0.5 part of polyhexamethylene guanide, water 50
~100 parts, 0.01~0.05 part of berberine, 0.1~0.5 part of sodium benzoate, 1~2 part of methanesulfonic acid, 0.1~0.5 part of octyl trichlamide,
1~2 part of lysozyme, 2~3 parts of lywallzyme, 0.1~0.5 part of astragalus polyose, 1~2 part of plant extraction liquid；Wherein, plant extraction liquid
Preparation method be：It is 5~10 parts of Chinese scholartree root, 8~10 parts of Japanese Stephania Root, 5~10 parts of elecampane by parts by weight, is divided with quality
Number is extracted twice for 70% ethanol solution, obtains medicinal extract, adds the honey that parts by weight are 1~5 part, convert parts by weight for 50~
100 parts of water, produces plant extraction liquid.
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| CN113711867A (en) * | 2021-09-24 | 2021-11-30 | 宁波市农业科学研究院 | Method for improving seed production yield of wild watermelon |
| CN114698521A (en) * | 2022-04-22 | 2022-07-05 | 海南茗卉农林科技发展有限公司 | Transplanting domestication and growth promotion method for tissue culture seedlings of color-leaf plants |
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| 陈继富: "无籽罗汉果的组织培养和快速繁殖", 《植物生理学报》 * |
| 黄春梅 等: "罗汉果组织培养与快繁研究进展", 《亚热带农业研究》 * |
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| Publication number | Priority date | Publication date | Assignee | Title |
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| CN110447414A (en) * | 2019-08-22 | 2019-11-15 | 桂林莱茵生物科技股份有限公司 | A method of improving Momordica grosvenori mogroside V content |
| CN110447414B (en) * | 2019-08-22 | 2022-04-05 | 桂林莱茵生物科技股份有限公司 | Method for increasing content of mogroside V in momordica grosvenori |
| CN113711867A (en) * | 2021-09-24 | 2021-11-30 | 宁波市农业科学研究院 | Method for improving seed production yield of wild watermelon |
| CN114698521A (en) * | 2022-04-22 | 2022-07-05 | 海南茗卉农林科技发展有限公司 | Transplanting domestication and growth promotion method for tissue culture seedlings of color-leaf plants |
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