CN106718920A - The method for improving momordica glycoside V content - Google Patents
The method for improving momordica glycoside V content Download PDFInfo
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- A—HUMAN NECESSITIES
- 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
- A01H4/001—Culture apparatus for tissue culture
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
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- A—HUMAN NECESSITIES
- 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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Abstract
The invention discloses a kind of method for improving momordica glycoside V content, comprise the following steps：Step one, methyl jasmonate is dissolved in the ethanol water that volume fraction is 20%, is configured to the ethanol mother liquor of the methyl jasmonate that concentration is 50mmol/L；Methyl jasmonate is dissolved in ethanol, it is 50~400 μm of methyl jasmonate solution of ol/L to be configured to concentration；Step 2, Luohanguo With Plantlets of Tissue Culture is seeded in solid medium, wherein, the ethanol mother liquor of methyl jasmonate is added in solid medium when solid medium is made, the ultimate density for making methyl jasmonate is 50~400 μm of ol/L；Step 3, by tissue culture transplantation of seedlings to field, 50~60d after Pollination of Luohanguo, daily it is early, middle and late it is each sprinkling methyl jasmonate solution, being dripped to blade face, continuously spray 10d.The present invention can effectively improve the yield of momordica glycoside V, with features such as instant effect, low cost, simple, convenient implementation, product nontoxic residue-frees, can meet the large-scale production of momordica glycoside V.
Description
Technical field
The present invention relates to a kind of implantation methods of Momordica grosvenori.It is more particularly related to a kind of improve sweet Momordica grosvenori
The method of glycosides V content.
Background technology
Momordica grosvenori is for the distinctive preciousness of China is medicinal and sweetener plant.Its fruit is cool in nature, sweet, with clearing heat and moistening lung, profit
Pharynx is opened the effects such as sound, laxation defaecation and anticancer, and the sweet glycosides V of active component is one of non-saccharide sweet substance most strong in the world, is sugarcane
300~400 times of sugared sugariness, are widely used in food, health products and medicine, are that diabetes patient, overweight people and hypertension are suffered from
The preferable sugar substitute of person.However, sweet glycosides V is only existed account in pulp of the fruit weight less than 15%, and content is only 1% or so, sternly
The sound development of Momordica grosvenori industry is constrained again.
Methyl jasmonate as plant signal elicitor can safely and effectively excite Plant Secondary Metabolites generation and
Accumulation.It is therefore preferable that suitable methyl jasmonate time of application and concentration, can improve to greatest extent medicinal plant activity into
The content divided.The report that methyl jasmonate improves momordica glycoside V content is there are no in the prior art.
The content of the invention
It is an object of the invention to provide a kind of method for improving momordica glycoside V content, it can effectively improve Momordica grosvenori
The yield of sweet glycosides V, with features such as instant effect, low cost, simple, convenient implementation, product nontoxic residue-frees, can meet sieve
The large-scale production of the sweet glycosides V of Chinese fruit.
In order to realize these purposes of the invention and further advantage, there is provided one kind improves momordica glycoside V content
Method, comprise the following steps：
Step one, methyl jasmonate is dissolved in the ethanol water that volume fraction is 20%, being configured to concentration is
The ethanol mother liquor of the methyl jasmonate of 50mmol/L, it is standby；Methyl jasmonate is dissolved in ethanol, be configured to concentration for 50~
400 μm of methyl jasmonate solution of ol/L, it is standby；
Step 2, Luohanguo With Plantlets of Tissue Culture is seeded in solid medium, is 60~66%, intensity of illumination in relative humidity
For 1400lux, light application time be 8h/d, cultivation temperature be 23 ± 2 DEG C under conditions of cultivate 30d, wherein, the system of solid medium
It is as method：The ethanol mother liquor of obtained methyl jasmonate in step one is added to solid culture when solid medium is made
In base, the ultimate density to methyl jasmonate is cooled to 24~26 DEG C after 50~400 μm of ol/L, solid medium to be sterilized；
Solid medium is also included：MS, 1.5mg/L 6- benzyls aminoadenine, 0.3mg/L indolebutyric acids, 3.5g/L agar, 30g/L sugarcanes
Sugar and 1.0g/L activated carbons；
Step 3, by step 2 cultivate after tissue culture transplantation of seedlings to field, 50~60d after Pollination of Luohanguo, daily
Obtained methyl jasmonate solution in step one of early, middle and late each sprinkling, being dripped to blade face, continuously sprays 10d.
Preferably, in the method for described raising momordica glycoside V content, in prepared jasmonic first in the step one
After the ethanol mother liquor of ester, also including being sterilized to the ethanol mother liquor of methyl jasmonate with the miillpore filter that aperture is 0.22 μm
Step.
Preferably, in the method for described raising momordica glycoside V content, by methyl jasmonate in the step 2
It is 2 to mass ratio is added in solid medium after ethanol mother liquor is added in solid medium:1 water-loss reducer and oyster shell powder,
In every liter of solid medium, the gross mass of water-loss reducer and oyster shell powder is 30~50g, when solid medium solidifies, water-loss reducer
In oyster shell powder all uniform immersion solid medium.
Preferably, in the method for described raising momordica glycoside V content, by Luohanguo With Plantlets of Tissue Culture in the step 2
After being seeded in solid medium, daily to the bactericide that the uniform spray temperature in surface of solid medium is 40~45 DEG C, institute
State after bactericide is mixed by the component of following parts by weight and be made：1~2 part of lysozyme, 2~3 parts of lywallzyme, pyrolkigneous liquid 200~
5~10 parts of 4~5 parts of 300 times of liquid and water.
The present invention at least includes following beneficial effect：
The present invention applies methyl jasmonate by Luohanguo With Plantlets of Tissue Culture and plantation Momordica grosvenori, to induce mogroside
The expression high of V key enzymes (such as glucosyltransferase and cytochrome P 450 enzymes) gene, so that quick in a short time
Improve the content of momordica glycoside V.
Because solid medium desiccation is very fast, metabolite is easily piled up on culture periphery, cause the nutrition of culture medium into
Divide skewness, be also unfavorable for that culture absorbs active ingredient.The present invention adds water-loss reducer and male in solid medium
Oyster shells, water-loss reducer is a kind of especially strong functional high molecule material of water absorbing capacity, when solid medium does not solidify, can be inhaled
The active ingredient in culture medium is received, the oyster shell powder after activation has microcellular structure, can also adsorb effective in many culture mediums
Composition, so can effectively keep nutritional ingredient, make the distribution of nutritional ingredient than more uniform, be absorbed by water-loss reducer and oyster shell powder
Nutritional ingredient with absorption can be slowly released, and be conducive to culture to absorb active ingredient longer, so as to promote arhat
The growth of fruit tissue-cultured seedling, is conducive to improving the content of momordica glycoside V.
After be seeded in Luohanguo With Plantlets of Tissue Culture in solid medium by the present invention, uniformly sprayed to the surface of solid medium or
Bactericide is smeared, wherein, the cell membrane of lysozyme energy hydrolytic bacteria makes bacterolysis, can make after being combined with virus virally inactivated；
Lywallzyme can hydrolyze the cell membrane of fungi, inactivate fungi；Pyrolkigneous liquid has the effects, bactericide such as sterilized, promotion plant growth
Barrier action can be played, can prevent the bacterium in culture environment, virus and fungi from influenceing the growth of Luohanguo With Plantlets of Tissue Culture, can also dropped
The low requirement to culture environment.
Easy to operate, low cost of the invention, it is environmentally friendly, it is suitable to large-scale production, with stronger practicality and push away
Wide value.
Further advantage of the invention, target and feature embody 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.
Specific 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 technique described in following embodiments, unless otherwise specified, is conventional method, institute
Reagent and material are stated, unless otherwise specified, is commercially obtained.
Embodiment 1
The present invention provides a kind of method for improving momordica glycoside V content, comprises the following steps：
Step one, methyl jasmonate is dissolved in the ethanol water that volume fraction is 20%, being configured to concentration is
The ethanol mother liquor of the methyl jasmonate of 50mmol/L, it is standby；Methyl jasmonate is dissolved in ethanol, concentration is configured to for 50 μ
The methyl jasmonate solution of mol/L, it is standby；
Step 2, Luohanguo With Plantlets of Tissue Culture is seeded in solid medium, is that 60%, intensity of illumination is in relative humidity
1400lux, light application time be 8h/d, cultivation temperature be 21 DEG C under conditions of cultivate 30d, wherein, the making side of solid medium
Method is：The ethanol mother liquor of obtained methyl jasmonate in step one is added to solid medium when solid medium is made
In, the ultimate density to methyl jasmonate is cooled to 24 DEG C after 50 μm of ol/L, solid medium to be sterilized, that is, be down to room temperature
Solidify solid medium；Solid medium is also included：MS, 1.5mg/L 6- benzyls aminoadenine, 0.3mg/L indolebutyric acids,
3.5g/L agar, 30g/L sucrose and 1.0g/L activated carbons；I.e. when solid medium is made, by jasmonic before agar solidification
The ethanol mother liquor of methyl esters is added in solid medium, is well mixed, and methyl jasmonate can be uniformly present in after agar solidification
In solid medium.
Step 3, will be cultivated in step 2 after tissue culture transplantation of seedlings to field, the 50d after Pollination of Luohanguo, it is early daily,
In, evening respectively sprinkling step one in obtained methyl jasmonate solution, being dripped to blade face, continuously spray 10d, treat carpopodium
It is changed into being sampled when yellowish-brown, pericarp change into faint yellow, momordica glycoside V content detection is carried out using HPLC methods.
In the method for described raising momordica glycoside V content, the ethanol of methyl jasmonate is being obtained in the step one
After mother liquor, also including being sterilized to the ethanol mother liquor of methyl jasmonate with the miillpore filter that aperture is 0.22 μm the step of.Energy
The ethanol mother liquor Carried bacteria of methyl jasmonate is prevented, the growth of tissue-cultured seedling is influenceed.
In the method for described raising momordica glycoside V content, by the ethanol mother liquor of methyl jasmonate in the step 2
It is 2 to mass ratio is added in solid medium before agar solidification after being added in solid medium:1 water-loss reducer and oyster shell
Powder, in every liter of solid medium, the gross mass of water-loss reducer and oyster shell powder is 30g, when solid medium solidify, water-loss reducer with
Oyster shell powder is all in uniform immersion solid medium.Because solid medium desiccation is very fast, in generation, is easily piled up on culture periphery
Thank to product, cause the nutrition distribution pattern of culture medium uneven, be also unfavorable for that culture absorbs active ingredient.In solid medium
In add water-loss reducer and oyster shell powder, water-loss reducer is a kind of especially strong functional high molecule material of water absorbing capacity, solid training
When foster base does not solidify, the active ingredient in culture medium can be absorbed, the oyster shell powder after activation has microcellular structure, can also adsorbed
Active ingredient in many culture mediums, so can effectively keep nutritional ingredient, make the distribution of nutritional ingredient than more uniform, be protected
Aqua and oyster shell powder are absorbed and the nutritional ingredient of absorption can be slowly released, be conducive to culture absorb longer effectively into
Point, so as to promote the growth of Luohanguo With Plantlets of Tissue Culture, be conducive to improving the content of momordica glycoside V.
In the method for described raising momordica glycoside V content, Luohanguo With Plantlets of Tissue Culture is seeded in admittedly in the step 2
After in body culture medium, daily to the bactericide that the uniform spray temperature in surface of solid medium is 40 DEG C, the bactericide by with
It is made after the component mixing of lower parts by weight：5 parts of 1 part of lysozyme, 2 parts of lywallzyme, 200 times of pyrolkigneous liquid, 4 parts of liquid and water.
After Luohanguo With Plantlets of Tissue Culture is seeded in solid medium, is uniformly sprayed to the surface of solid medium or painting is obliterated
Microbial inoculum, wherein, the cell membrane of lysozyme energy hydrolytic bacteria makes bacterolysis, can make after being combined with virus virally inactivated；Lywallzyme
The cell membrane of fungi can be hydrolyzed, fungi is inactivated；Pyrolkigneous liquid has the effects such as sterilized, promotion plant growth, and bactericide can be played
Barrier action, can prevent the bacterium in culture environment, virus and fungi from influenceing the growth of Luohanguo With Plantlets of Tissue Culture, can also reduce to training
Support the requirement of environment.
Embodiment 2
The present invention provides a kind of method for improving momordica glycoside V content, comprises the following steps：
Step one, methyl jasmonate is dissolved in the ethanol water that volume fraction is 20%, being configured to concentration is
The ethanol mother liquor of the methyl jasmonate of 50mmol/L, it is standby；Methyl jasmonate is dissolved in ethanol, concentration is configured to for 400 μ
The methyl jasmonate solution of mol/L, it is standby；
Step 2, Luohanguo With Plantlets of Tissue Culture is seeded in solid medium, is that 66%, intensity of illumination is in relative humidity
1400lux, light application time be 8h/d, cultivation temperature be 25 DEG C under conditions of cultivate 30d, wherein, the making side of solid medium
Method is：The ethanol mother liquor of obtained methyl jasmonate in step one is added to solid medium when solid medium is made
In, the ultimate density to methyl jasmonate is cooled to 26 DEG C after 400 μm of ol/L, solid medium to be sterilized, that is, be down to room temperature
Solidify solid medium；Solid medium is also included：MS, 1.5mg/L 6- benzyls aminoadenine, 0.3mg/L indolebutyric acids,
3.5g/L agar, 30g/L sucrose and 1.0g/L activated carbons；I.e. when solid medium is made, by jasmonic before agar solidification
The ethanol mother liquor of methyl esters is added in solid medium, is well mixed, and methyl jasmonate can be uniformly present in after agar solidification
In solid medium.
Step 3, will be cultivated in step 2 after tissue culture transplantation of seedlings to field, the 60d after Pollination of Luohanguo, it is early daily,
In, evening respectively sprinkling step one in obtained methyl jasmonate solution, being dripped to blade face, continuously spray 10d, treat carpopodium
It is changed into being sampled when yellowish-brown, pericarp change into faint yellow, momordica glycoside V content detection is carried out using HPLC methods.
In the method for described raising momordica glycoside V content, the ethanol of methyl jasmonate is being obtained in the step one
After mother liquor, also including being sterilized to the ethanol mother liquor of methyl jasmonate with the miillpore filter that aperture is 0.22 μm the step of.Energy
The ethanol mother liquor Carried bacteria of methyl jasmonate is prevented, the growth of tissue-cultured seedling is influenceed.
In the method for described raising momordica glycoside V content, by the ethanol mother liquor of methyl jasmonate in the step 2
It is 2 to mass ratio is added in solid medium after being added in solid medium:1 water-loss reducer and oyster shell powder, every liter of solid
In culture medium, the gross mass of water-loss reducer and oyster shell powder is 50g, and when solid medium solidifies, water-loss reducer and oyster shell powder are complete
Portion is uniformly immersed in solid medium.Because solid medium desiccation is very fast, metabolite is easily piled up on culture periphery, causes
The nutrition distribution pattern of culture medium is uneven, is also unfavorable for that culture absorbs active ingredient.Guarantor is added in solid medium
Aqua and oyster shell powder, water-loss reducer are a kind of especially strong functional high molecule materials of water absorbing capacity, are not coagulated in solid medium
Gu when, the active ingredient in culture medium can be absorbed, the oyster shell powder after activation has microcellular structure, can also adsorb many culture mediums
In active ingredient, so can effectively keep nutritional ingredient, make the distribution of nutritional ingredient than more uniform, by water-loss reducer and oyster
Shell powder is absorbed and the nutritional ingredient of absorption can be slowly released, and is conducive to culture to absorb active ingredient longer, so as to
Promote the growth of Luohanguo With Plantlets of Tissue Culture, be conducive to improving the content of momordica glycoside V.
In the method for described raising momordica glycoside V content, Luohanguo With Plantlets of Tissue Culture is seeded in admittedly in the step 2
After in body culture medium, daily to the bactericide that the uniform spray temperature in surface of solid medium is 45 DEG C, the bactericide by with
It is made after the component mixing of lower parts by weight：10 parts of 2 parts of lysozyme, 3 parts of lywallzyme, 300 times of pyrolkigneous liquid, 5 parts of liquid and water.
After Luohanguo With Plantlets of Tissue Culture is seeded in solid medium, is uniformly sprayed to the surface of solid medium or painting is obliterated
Microbial inoculum, wherein, the cell membrane of lysozyme energy hydrolytic bacteria makes bacterolysis, can make after being combined with virus virally inactivated；Lywallzyme
The cell membrane of fungi can be hydrolyzed, fungi is inactivated；Pyrolkigneous liquid has the effects such as sterilized, promotion plant growth, and bactericide can be played
Barrier action, can prevent the bacterium in culture environment, virus and fungi from influenceing the growth of Luohanguo With Plantlets of Tissue Culture, can also reduce to training
Support the requirement of environment.
Embodiment 3
The present invention provides a kind of method for improving momordica glycoside V content, comprises the following steps：
Step one, methyl jasmonate is dissolved in the ethanol water that volume fraction is 20%, being configured to concentration is
The ethanol mother liquor of the methyl jasmonate of 50mmol/L, it is standby；Methyl jasmonate is dissolved in ethanol, concentration is configured to for 220 μ
The methyl jasmonate solution of mol/L, it is standby；
Step 2, Luohanguo With Plantlets of Tissue Culture is seeded in solid medium, is that 63%, intensity of illumination is in relative humidity
1400lux, light application time be 8h/d, cultivation temperature be 23 DEG C under conditions of cultivate 30d, wherein, the making side of solid medium
Method is：The ethanol mother liquor of obtained methyl jasmonate in step one is added to solid medium when solid medium is made
In, the ultimate density to methyl jasmonate is cooled to 25 DEG C after 220 μm of ol/L, solid medium to be sterilized, that is, be down to room temperature
Solidify solid medium；Solid medium is also included：MS, 1.5mg/L 6- benzyls aminoadenine, 0.3mg/L indolebutyric acids,
3.5g/L agar, 30g/L sucrose and 1.0g/L activated carbons；I.e. when solid medium is made, by jasmonic before agar solidification
The ethanol mother liquor of methyl esters is added in solid medium, is well mixed, and methyl jasmonate can be uniformly present in after agar solidification
In solid medium.
Step 3, will be cultivated in step 2 after tissue culture transplantation of seedlings to field, the 55d after Pollination of Luohanguo, it is early daily,
In, evening respectively sprinkling step one in obtained methyl jasmonate solution, being dripped to blade face, continuously spray 10d, treat carpopodium
It is changed into being sampled when yellowish-brown, pericarp change into faint yellow, momordica glycoside V content detection is carried out using HPLC methods.
In the method for described raising momordica glycoside V content, the ethanol of methyl jasmonate is being obtained in the step one
After mother liquor, also including being sterilized to the ethanol mother liquor of methyl jasmonate with the miillpore filter that aperture is 0.22 μm the step of.Energy
The ethanol mother liquor Carried bacteria of methyl jasmonate is prevented, the growth of tissue-cultured seedling is influenceed.
In the method for described raising momordica glycoside V content, by the ethanol mother liquor of methyl jasmonate in the step 2
It is 2 to mass ratio is added in solid medium after being added in solid medium:1 water-loss reducer and oyster shell powder, every liter of solid
In culture medium, the gross mass of water-loss reducer and oyster shell powder is 40g, and when solid medium solidifies, water-loss reducer and oyster shell powder are complete
Portion is uniformly immersed in solid medium.Because solid medium desiccation is very fast, metabolite is easily piled up on culture periphery, causes
The nutrition distribution pattern of culture medium is uneven, is also unfavorable for that culture absorbs active ingredient.Guarantor is added in solid medium
Aqua and oyster shell powder, water-loss reducer are a kind of especially strong functional high molecule materials of water absorbing capacity, are not coagulated in solid medium
Gu when, the active ingredient in culture medium can be absorbed, the oyster shell powder after activation has microcellular structure, can also adsorb many culture mediums
In active ingredient, so can effectively keep nutritional ingredient, make the distribution of nutritional ingredient than more uniform, by water-loss reducer and oyster
Shell powder is absorbed and the nutritional ingredient of absorption can be slowly released, and is conducive to culture to absorb active ingredient longer, so as to
Promote the growth of Luohanguo With Plantlets of Tissue Culture, be conducive to improving the content of momordica glycoside V.
In the method for described raising momordica glycoside V content, Luohanguo With Plantlets of Tissue Culture is seeded in admittedly in the step 2
After in body culture medium, daily to the bactericide that the uniform spray temperature in surface of solid medium is 42 DEG C, the bactericide by with
It is made after the component mixing of lower parts by weight：7 parts of 2 parts of lysozyme, 2 parts of lywallzyme, 250 times of pyrolkigneous liquid, 4 parts of liquid and water.
After Luohanguo With Plantlets of Tissue Culture is seeded in solid medium, is uniformly sprayed to the surface of solid medium or painting is obliterated
Microbial inoculum, wherein, the cell membrane of lysozyme energy hydrolytic bacteria makes bacterolysis, can make after being combined with virus virally inactivated；Lywallzyme
The cell membrane of fungi can be hydrolyzed, fungi is inactivated；Pyrolkigneous liquid has the effects such as sterilized, promotion plant growth, and bactericide can be played
Barrier action, can prevent the bacterium in culture environment, virus and fungi from influenceing the growth of Luohanguo With Plantlets of Tissue Culture, can also reduce to training
Support the requirement of environment.
Comparative example 1
A kind of implantation methods of Momordica grosvenori of the present invention, comprise the following steps：
Step one, Luohanguo With Plantlets of Tissue Culture is seeded in solid medium, is that 60%, intensity of illumination is in relative humidity
1400lux, light application time are 8h/d, cultivation temperature to cultivate 30d under conditions of 21 DEG C；Solid medium is included：MS、1.5mg/
L 6- benzyls aminoadenine, 0.3mg/L indolebutyric acids, 3.5g/L agar, 30g/L sucrose and 1.0g/L activated carbons, do not contain jasmine
Jasmine acid methyl esters；
Step 3, by step 2 cultivate after tissue culture transplantation of seedlings to field, planted according to a conventional method after transplanting, other training
The condition of supporting is identical with embodiment 1~3, but does not spray methyl jasmonate solution.
In embodiment 1~3 and comparative example 1, it is changed into being sampled when yellowish-brown, pericarp change into faint yellow whne carpopodium, using HPLC
Method carries out momordica glycoside V content detection, the results are shown in Table 1, and sweet glycosides V content refers to that sweet glycosides V content accounts for the percentage of fruit content in table 1
Number.
The testing result of the embodiment 1~3 of table 1 and the momordica glycoside V content of comparative example 1
| Embodiment | Embodiment 1 | Embodiment 2 | Embodiment 3 | Comparative example 1 |
| Sweet glycosides V content | 1.04% | 1.35% | 1.16% | 0.77% |
As shown in Table 1, using the method for embodiment 1~3 compared with the method for comparative example 1, embodiment 1~3 can be significantly
The content of momordica glycoside V is improved, illustrates that applying methyl jasmonate can be effectively in Luohanguo With Plantlets of Tissue Culture and plantation Momordica grosvenori
The content of momordica glycoside V is improved, the method instant effect, low cost, simple, convenient implementation can be momordica glycoside V
Large-scale production provides new approaches.
Although embodiment of the present invention is disclosed as above, it is not restricted to listed in specification and implementation method
With, it can be applied to various suitable the field of the invention completely, for those skilled in the art, can be easily
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 (4)
1. it is a kind of improve momordica glycoside V content method, it is characterised in that comprise the following steps：
Step one, methyl jasmonate is dissolved in the ethanol water that volume fraction is 20%, is configured to concentration for 50mmol/L
Methyl jasmonate ethanol mother liquor, it is standby；Methyl jasmonate is dissolved in ethanol, concentration is configured to for 50~400 μm of ol/L
Methyl jasmonate solution, it is standby；
Step 2, Luohanguo With Plantlets of Tissue Culture is seeded in solid medium, is that 60~66%, intensity of illumination is in relative humidity
1400lux, light application time be 8h/d, cultivation temperature be 23 ± 2 DEG C under conditions of cultivate 30d, wherein, make solid medium
When the ethanol mother liquor of obtained methyl jasmonate in step one is added in solid medium, it is final dense to methyl jasmonate
After spending for 50~400 μm of ol/L, solid medium is sterilized, be cooled to 24~26 DEG C；Solid medium is also included：MS、
1.5mg/L 6- benzyls aminoadenine, 0.3mg/L indolebutyric acids, 3.5g/L agar, 30g/L sucrose and 1.0g/L activated carbons；
Step 3, will be cultivated in step 2 after tissue culture transplantation of seedlings to field, 50~60d after Pollination of Luohanguo, it is early daily,
In, evening respectively sprinkling step one in obtained methyl jasmonate solution, being dripped to blade face, continuously spray 10d.
2. the method for improving momordica glycoside V content as claimed in claim 1, it is characterised in that in system in the step one
After the ethanol mother liquor of methyl jasmonate, also including with the miillpore filter that aperture is 0.22 μm to the ethanol mother liquor of methyl jasmonate
The step of being sterilized.
3. the method for improving momordica glycoside V content as claimed in claim 1, it is characterised in that by jasmine in the step 2
It is 2 to mass ratio is added in solid medium after the ethanol mother liquor of jasmine acid methyl esters is added in solid medium:1 water-loss reducer
And oyster shell powder, in every liter of solid medium, the gross mass of water-loss reducer and oyster shell powder is 30~50g, when solid medium is solidifying
Gu when, water-loss reducer and oyster shell powder are all uniformly immersed in solid medium.
4. the method for improving momordica glycoside V content as claimed in claim 1, it is characterised in that by sieve in the step 2
It it is daily 40~45 DEG C to the uniform spray temperature in surface of solid medium after Chinese fruit tissue culture plant inoculation is in the solid medium
Bactericide, the bactericide is made after being mixed by the component of following parts by weight：1~2 part of lysozyme, 2~3 parts of lywallzyme, wood
5~10 parts of 200~300 times of vinegar liquid, 4~5 parts of liquid and water.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201611247647.XA CN106718920B (en) | 2016-12-29 | 2016-12-29 | The method for improving momordica glycoside V content |
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| CN107189976A (en) * | 2017-07-28 | 2017-09-22 | 李华政 | Promote the method for Momordica grosvenori UGT72B21 gene expressions |
| CN107223568A (en) * | 2017-07-28 | 2017-10-03 | 聂天军 | Promote the method for Momordica grosvenori CYP89A143 gene expressions |
| CN107232065A (en) * | 2017-07-28 | 2017-10-10 | 李华政 | Promote the method for Momordica grosvenori UGT84A21 gene expressions |
| CN107232064A (en) * | 2017-07-28 | 2017-10-10 | 黄小华 | Promote the method for Momordica grosvenori CYP23 gene expressions |
| CN107232063A (en) * | 2017-07-28 | 2017-10-10 | 黄小华 | Promote the method for Momordica grosvenori CYP24 gene expressions |
| CN107251839A (en) * | 2017-07-28 | 2017-10-17 | 韦荣昌 | Promote the method for Momordica grosvenori UGT71V1 gene expressions |
| CN107258541A (en) * | 2017-07-28 | 2017-10-20 | 韦荣昌 | Promote the method for Momordica grosvenori UGT7 gene expressions |
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| CN107295971A (en) * | 2017-07-28 | 2017-10-27 | 韦荣昌 | Promote the method for Momordica grosvenori CYP4 gene expressions |
| CN107306792A (en) * | 2017-07-28 | 2017-11-03 | 聂天军 | Promote the method for Momordica grosvenori CYP87D17 gene expressions |
| CN107318652A (en) * | 2017-07-28 | 2017-11-07 | 韦荣昌 | Promote the method for Momordica grosvenori HMGR gene expressions |
| CN107318651A (en) * | 2017-07-28 | 2017-11-07 | 李华政 | Promote the method for Momordica grosvenori UGT2 gene expressions |
| CN107318654A (en) * | 2017-07-28 | 2017-11-07 | 韦荣昌 | Promote the method for Momordica grosvenori CS gene expressions |
| CN107360971A (en) * | 2017-07-28 | 2017-11-21 | 韦荣昌 | Promote the method for Momordica grosvenori SS gene expressions |
| CN107360970A (en) * | 2017-07-28 | 2017-11-21 | 李华政 | Promote the method for Momordica grosvenori UGT74AC1 gene expressions |
| CN107372109A (en) * | 2017-07-28 | 2017-11-24 | 李华政 | Promote the method for Momordica grosvenori UGT73AF1 gene expressions |
| CN107432246A (en) * | 2017-07-28 | 2017-12-05 | 黄小华 | Promote the method for Momordica grosvenori CYP43 gene expressions |
| CN109220656A (en) * | 2018-11-22 | 2019-01-18 | 衡阳仕杰农业发展有限公司 | A kind of implantation methods of selenium-rich rice |
| CN109757381A (en) * | 2019-03-20 | 2019-05-17 | 丽江海贝瑞生物科技有限公司 | A kind of bletilla striata tissue culture and rapid propagation method |
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| CN107318652A (en) * | 2017-07-28 | 2017-11-07 | 韦荣昌 | Promote the method for Momordica grosvenori HMGR gene expressions |
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| CN107189976A (en) * | 2017-07-28 | 2017-09-22 | 李华政 | Promote the method for Momordica grosvenori UGT72B21 gene expressions |
| CN107232064A (en) * | 2017-07-28 | 2017-10-10 | 黄小华 | Promote the method for Momordica grosvenori CYP23 gene expressions |
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| CN107223568A (en) * | 2017-07-28 | 2017-10-03 | 聂天军 | Promote the method for Momordica grosvenori CYP89A143 gene expressions |
| CN107258542A (en) * | 2017-07-28 | 2017-10-20 | 聂天军 | Promote the method for Momordica grosvenori CYP749A36 gene expressions |
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| CN107360970A (en) * | 2017-07-28 | 2017-11-21 | 李华政 | Promote the method for Momordica grosvenori UGT74AC1 gene expressions |
| CN107372109A (en) * | 2017-07-28 | 2017-11-24 | 李华政 | Promote the method for Momordica grosvenori UGT73AF1 gene expressions |
| CN107432246A (en) * | 2017-07-28 | 2017-12-05 | 黄小华 | Promote the method for Momordica grosvenori CYP43 gene expressions |
| WO2019136924A1 (en) * | 2018-01-15 | 2019-07-18 | 华东理工大学 | Method for promoting suspension cell growth of momordica grosvenori and improving content of mogroside v by means of salt stress |
| CN110684708A (en) * | 2018-07-06 | 2020-01-14 | 华东理工大学 | Method for amplifying and culturing fructus momordicae suspension cells |
| CN110684708B (en) * | 2018-07-06 | 2023-05-16 | 华东理工大学 | Method for amplifying and culturing momordica grosvenori suspension cells |
| CN109220656A (en) * | 2018-11-22 | 2019-01-18 | 衡阳仕杰农业发展有限公司 | A kind of implantation methods of selenium-rich rice |
| CN109757381A (en) * | 2019-03-20 | 2019-05-17 | 丽江海贝瑞生物科技有限公司 | A kind of bletilla striata tissue culture and rapid propagation method |
| CN109757381B (en) * | 2019-03-20 | 2022-03-08 | 丽江海贝瑞生物科技有限公司 | Bletilla striata tissue culture and rapid propagation method |
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