CN106797974A - The method for improving Simon glycosides I contents in Momordica grosvenori - Google Patents
The method for improving Simon glycosides I contents in Momordica grosvenori Download PDFInfo
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- CN106797974A CN106797974A CN201611249215.2A CN201611249215A CN106797974A CN 106797974 A CN106797974 A CN 106797974A CN 201611249215 A CN201611249215 A CN 201611249215A CN 106797974 A CN106797974 A CN 106797974A
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- momordica grosvenori
- methyl jasmonate
- contents
- simon glycosides
- tissue culture
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01N—PRESERVATION OF BODIES OF HUMANS OR ANIMALS OR PLANTS OR PARTS THEREOF; BIOCIDES, e.g. AS DISINFECTANTS, AS PESTICIDES OR AS HERBICIDES; PEST REPELLANTS OR ATTRACTANTS; PLANT GROWTH REGULATORS
- A01N65/00—Biocides, pest repellants or attractants, or plant growth regulators containing material from algae, lichens, bryophyta, multi-cellular fungi or plants, or extracts thereof
-
- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01G—HORTICULTURE; CULTIVATION OF VEGETABLES, FLOWERS, RICE, FRUIT, VINES, HOPS OR SEAWEED; FORESTRY; WATERING
- A01G22/00—Cultivation of specific crops or plants not otherwise provided for
-
- 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
-
- 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
- A01H4/008—Methods for regeneration to complete plants
-
- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01N—PRESERVATION OF BODIES OF HUMANS OR ANIMALS OR PLANTS OR PARTS THEREOF; BIOCIDES, e.g. AS DISINFECTANTS, AS PESTICIDES OR AS HERBICIDES; PEST REPELLANTS OR ATTRACTANTS; PLANT GROWTH REGULATORS
- A01N37/00—Biocides, pest repellants or attractants, or plant growth regulators containing organic compounds containing a carbon atom having three bonds to hetero atoms with at the most two bonds to halogen, e.g. carboxylic acids
- A01N37/42—Biocides, pest repellants or attractants, or plant growth regulators containing organic compounds containing a carbon atom having three bonds to hetero atoms with at the most two bonds to halogen, e.g. carboxylic acids containing within the same carbon skeleton a carboxylic group or a thio analogue, or a derivative thereof, and a carbon atom having only two bonds to hetero atoms with at the most one bond to halogen, e.g. keto-carboxylic acids
-
- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01N—PRESERVATION OF BODIES OF HUMANS OR ANIMALS OR PLANTS OR PARTS THEREOF; BIOCIDES, e.g. AS DISINFECTANTS, AS PESTICIDES OR AS HERBICIDES; PEST REPELLANTS OR ATTRACTANTS; PLANT GROWTH REGULATORS
- A01N61/00—Biocides, pest repellants or attractants, or plant growth regulators containing substances of unknown or undetermined composition, e.g. substances characterised only by the mode of action
-
- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01N—PRESERVATION OF BODIES OF HUMANS OR ANIMALS OR PLANTS OR PARTS THEREOF; BIOCIDES, e.g. AS DISINFECTANTS, AS PESTICIDES OR AS HERBICIDES; PEST REPELLANTS OR ATTRACTANTS; PLANT GROWTH REGULATORS
- A01N65/00—Biocides, pest repellants or attractants, or plant growth regulators containing material from algae, lichens, bryophyta, multi-cellular fungi or plants, or extracts thereof
- A01N65/40—Liliopsida [monocotyledons]
- A01N65/46—Stemonaceae [Stemona family], e.g. croomia
Abstract
The invention discloses a kind of method for improving Simon glycosides I contents in Momordica grosvenori, methyl jasmonate is applied in Momordica grosvenori tissue culture process or cultivation.The present invention can effectively improve the yield of Momordica grosvenori Simon glycosides I, with features such as instant effect, low cost, simple, convenient implementation, product nontoxic residue-frees, to meet the large-scale production of Momordica grosvenori Simon glycosides I, with stronger practicality and promotional value.
Description
Technical field
The present invention relates to plant biotechnology field.It is more particularly related to a kind of improve Simon in Momordica grosvenori
The method of glycosides I contents.
Background technology
Momordica grosvenori (Siraitia grosvenorii) is for the distinctive preciousness of China is medicinal and sweetener plant.Its fruit
Cool, sweet, with clearing heat and moistening lung, relieving sore-throat opens the effects such as sound, laxation defaecation and anticancer.Research shows that Momordica grosvenori Simon glycosides I is mesh
The non-saccharide sweet substance most strong in the world of preceding discovery, is 563 times of 5% sweetness of cane sugar in a ten thousandth concentration, is glycosuria
The preferable sugar substitute of patient, overweight people and hyperpietic.However, Simon glycosides I is only existed accounts for pulp of the fruit weight less than 15%
It is interior, and content is extremely inefficient, so that it is expensive, its extensive use in food, health products and medicine is greatly limit, sternly
The sound development of Momordica grosvenori industry is constrained again.
Universally present in plant, exogenous application can excite the expression of defence plant gene to methyl jasmonate, lure
The chemical defence of plant is led, the effect similar to mechanical damage and insect's food-taking is produced.Jasmonic first is there are no in the prior art
Ester improves the report of Simon glycosides I contents in Momordica grosvenori.
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 method for improving Simon glycosides I contents in Momordica grosvenori, it can be effective
Ground improves the yield of Momordica grosvenori Simon glycosides I, with instant effect, low cost, simple, convenient implementation, product nontoxic residue-free
Etc. feature, to meet the large-scale production of Momordica grosvenori Simon glycosides I, with stronger practicality and promotional value.
In order to realize these purposes of the invention and further advantage, there is provided glycosides I in Simon in one kind raising Momordica grosvenori
The method of content, methyl jasmonate is applied in Momordica grosvenori tissue culture process or cultivation.
Preferably, the described method for improving Simon glycosides I contents in Momordica grosvenori, during tissue culture, by Momordica grosvenori tissue culture
Seedling is seeded in the MS solid mediums containing methyl jasmonate, be placed in relative humidity 60~66%, intensity of illumination 1300~
28~31d is cultivated under the conditions of 1500Lux, light application time 8h/d, 21~25 DEG C of temperature, wherein, the addition of methyl jasmonate is
50~400 μm of ol/L.
Preferably, the described method for improving Simon glycosides I contents in Momordica grosvenori, in cultivation, by methyl jasmonate
It is 50~400 μm of methyl jasmonate solution of ol/L to be made into concentration, after 20~30d of Pollination of Luohanguo, jasmonic first is sprayed daily
In Momordica grosvenori surface, being dripped to surface, early, middle and late each sprinkling once, continuously sprays 8~12d to ester solution.
Preferably, the described method for improving Simon glycosides I contents in Momordica grosvenori, including：
Step one, Luohanguo With Plantlets of Tissue Culture is seeded in the MS solid mediums containing methyl jasmonate, be placed in relatively wet
28~31d is cultivated under the conditions of degree 60~66%, 1300~1500Lux of intensity of illumination, light application time 8h/d, 21~25 DEG C of temperature,
Wherein, the addition of methyl jasmonate is 50~400 μm of ol/L；
Step 2, step one is cultivated after tissue culture transplantation of seedlings to field, after 20~30d of Pollination of Luohanguo, daily sprinkling
Concentration be 50~400 μm of methyl jasmonate solution of ol/L in Momordica grosvenori surface, being dripped to surface, early, middle and late each sprinkling
Once, 8~12d is continuously sprayed.
Preferably, the described method for improving Simon glycosides I contents in Momordica grosvenori, with the ethanol water that volume fraction is 2%
Solution dissolves methyl jasmonate, is configured to the ethanol mother liquor that concentration is 50mmol/L methyl jasmonates, and with 0.22 μm of micropore
Filter membrane sterilizes, and the ethanol mother liquor of the methyl jasmonate after sterilizing is added in MS solid mediums, to methyl jasmonate concentration
It is 50~400 μm of ol/L, sterilizes and be cooled to 24~26 DEG C, wherein, MS solid mediums include：MS, 1.5mg/L6- benzyl amino
Adenine, 0.3mg/L indolebutyric acids, 3.5g/L agar, 30g/L sucrose and 1.0g/L activated carbons.
Preferably, the described method for improving Simon glycosides I contents in Momordica grosvenori, with the ethanol water that volume fraction is 2%
Solution dissolves methyl jasmonate, and it is 50~400 μm of methyl jasmonate solution of ol/L to be configured to concentration.
Preferably, the described method for improving Simon glycosides I contents in Momordica grosvenori, MS solid mediums also include regulation
Agent, the addition of conditioning agent is 5 μm of ol/L, and conditioning agent includes that mass ratio is 2:1:0.1 bamboo vinegar, tuber of stemona water extract, quercitrin
Element.
Preferably, the described method for improving Simon glycosides I contents in Momordica grosvenori, tissue culture process includes two benches, first
Stage cultivates 15d, and a MS solid medium is changed every 3~5d, and the MS solid mediums being replaced are heated to being molten into liquid
State mixture, sponge is immersed in liquefied mixture and is fully absorbed, and is then covered by above the MS solid mediums changed, directly
Remove and change when changing culture medium to next time；Second stage culture 15d, changes a MS solid medium, often every 5~7d
2d after secondary replacing carries out atomization process to the MS solid mediums for being inoculated with tissue-cultured seedling, and Alevaire is that mass fraction is 1%
Casein solution.
Preferably, the described method for improving Simon glycosides I contents in Momordica grosvenori, more by the top girdling of plant after pollination
Individual annular incision, two neighboring otch spacing 5mm, first sprinkling mass fraction is 1% flavonol solution and then uses peach gum daily
Smear otch, finally use cordate houttuynia bondage otch, until starting to spray methyl jasmonate solution.
The present invention at least includes following beneficial effect：
Firstth, the present invention applies methyl jasmonate by tissue culture and/or cultivation, can induce Momordica grosvenori Simon glycosides
The expression high of I key enzymes (such as glucosyltransferase and cytochrome P 450 enzymes) gene, so that quick in a short time
Improve the content of Momordica grosvenori Simon glycosides I；The applying concentration and time of application of tissue culture process or the preferred methyl jasmonate of cultivation,
By tissue culture plant inoculation in the MS solid mediums containing methyl jasmonate, or by methyl jasmonate spray solution in pollination 20~
Momordica grosvenori surface after 30d, the accumulation of Momordica grosvenori Simon glycosides I, can to greatest extent improve medicinal plant work in promotion nursery stock
The content of property composition；
Secondth, the present invention in MS solid mediums by adding the tune compounded by bamboo vinegar, tuber of stemona water extract, Quercetin
Section agent, auxiliary promotes to glucosyltransferase and the activity level of cytochrome P 450 enzymes Buchner's bodies, realizes secondary substance
Accumulation, tuber of stemona water extract also have insecticidal action, Quercetin improve conditioning agent Luohanguo With Plantlets of Tissue Culture adhesive force, bamboo vinegar
Promote Luohanguo With Plantlets of Tissue Culture growth, concentration proportioning avoids cumarin from precipitating；
3rd, the respectively different time interval of two benches changes culture medium, and supplement Momordica grosvenori grows different phase because of consumption
Required nutriment, first stage immersion sponge avoids used culture medium nutriment from wasting, while being covered in top
Sink to absorbing with water retention, nutriment, second stage utilizes casein solution atomization process, it is to avoid tissue-cultured seedling blade table
The loss of flour nutrition material, promotes absorption of the root to methyl jasmonate in culture medium；
4th, girdling after the pollination of cultivation stage, temporarily hinders organic substance to convey downwards, increases wound top nutrient product
Tired, sprinkling flavonol solution suppresses oozing out for biology enzyme, and peach gum smears otch and is beneficial to wound self-healing, and cordate houttuynia bondage otch is sterilized
Expelling parasite, forms defense structure, promotes absorption of the later stage to methyl jasmonate.
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 appreciated that it is used herein such as " have ", "comprising" and " including " term do not allot one or many
The presence or addition of individual other elements or its combination.
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>
Step one, with volume fraction be 2% ethanol water dissolving methyl jasmonate, be configured to concentration for 200mmol/
The ethanol mother liquor of L methyl jasmonates, and with 0.22 μm of miillpore filter sterilizing, by the ethanol mother liquor of the methyl jasmonate after sterilizing
It is added in MS solid mediums, the addition to methyl jasmonate is 200 μm of ol/L, sterilizes and be cooled to 24~26 DEG C, its
In, MS solid mediums include：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 2, Luohanguo With Plantlets of Tissue Culture is seeded in the MS solid mediums containing methyl jasmonate that step one is obtained
In, it is placed under the conditions of relative humidity 60~66%, 1300~1500Lux of intensity of illumination, light application time 8h/d, 21~25 DEG C of temperature
Culture 30d, then carries out conventional field culture by tissue culture transplantation of seedlings to field.
<Embodiment 2>
Methyl jasmonate is dissolved with the ethanol water that volume fraction is 2%, it is 200 μm of jasmines of ol/L to be configured to concentration
Sour methyl ester solution, after Pollination of Luohanguo 25d, sprays methyl jasmonate solution in Momordica grosvenori surface daily, being dripped to surface,
Early, middle and late each sprinkling once, continuously sprays 10d.
<Embodiment 3>
Step one, with volume fraction be 2% ethanol water dissolving methyl jasmonate, be configured to concentration for 50mmol/L
The ethanol mother liquor of methyl jasmonate, and with 0.22 μm of miillpore filter sterilizing, by the ethanol mother liquor of the methyl jasmonate after sterilizing
It is added in MS solid mediums, the addition to methyl jasmonate is 50 μm of ol/L, sterilizes and be cooled to 24~26 DEG C, its
In, MS solid mediums include：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 2, Luohanguo With Plantlets of Tissue Culture is seeded in the MS solid mediums containing methyl jasmonate that step one is obtained
In, it is placed under the conditions of relative humidity 60~66%, 1300~1500Lux of intensity of illumination, light application time 8h/d, 21~25 DEG C of temperature
Culture 28d；
Step 3, with volume fraction be 2% ethanol water dissolving methyl jasmonate, be configured to concentration for 50 μm of ol/L
Methyl jasmonate solution, the tissue culture transplantation of seedlings after step one is cultivated after Pollination of Luohanguo 20d, sprays dense daily to field
It is 50 μm of methyl jasmonate solution of ol/L in Momordica grosvenori surface to spend, being dripped to surface, early, middle and late each sprinkling once, even
It is continuous to spray 8d.
<Embodiment 4>
Step one, with volume fraction be 2% ethanol water dissolving methyl jasmonate, be configured to concentration for 400mmol/
The ethanol mother liquor of L methyl jasmonates, and with 0.22 μm of miillpore filter sterilizing, by the ethanol mother liquor of the methyl jasmonate after sterilizing
It is added in MS solid mediums, the addition to methyl jasmonate is 400 μm of ol/L, sterilizes and be cooled to 24~26 DEG C, its
In, MS solid mediums include：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 2, Luohanguo With Plantlets of Tissue Culture is seeded in the MS solid mediums containing methyl jasmonate that step one is obtained
In, it is placed under the conditions of relative humidity 60~66%, 1300~1500Lux of intensity of illumination, light application time 8h/d, 21~25 DEG C of temperature
Culture 31d；
Step 3, with volume fraction be 2% ethanol water dissolving methyl jasmonate, be configured to concentration for 400 μm of ol/
The methyl jasmonate solution of L, the tissue culture transplantation of seedlings after step one is cultivated after Pollination of Luohanguo 30d, is sprayed dense daily to field
It is 400 μm of methyl jasmonate solution of ol/L in Momordica grosvenori surface to spend, being dripped to surface, early, middle and late each sprinkling once,
Continuously spray 12d.
<Embodiment 5>
Step one, with volume fraction be 2% ethanol water dissolving methyl jasmonate, be configured to concentration for 200mmol/
The ethanol mother liquor of L methyl jasmonates, and with 0.22 μm of miillpore filter sterilizing, by the ethanol mother liquor of the methyl jasmonate after sterilizing
It is added in MS solid mediums, the addition to methyl jasmonate is 200 μm of ol/L, sterilizes and be cooled to 24~26 DEG C, its
In, MS solid mediums include：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 2, Luohanguo With Plantlets of Tissue Culture is seeded in the MS solid mediums containing methyl jasmonate that step one is obtained
In, it is placed under the conditions of relative humidity 60~66%, 1300~1500Lux of intensity of illumination, light application time 8h/d, 21~25 DEG C of temperature
Culture 30d；
Step 3, with volume fraction be 2% ethanol water dissolving methyl jasmonate, be configured to concentration for 200 μm of ol/
The methyl jasmonate solution of L, the tissue culture transplantation of seedlings after step one is cultivated after Pollination of Luohanguo 25d, is sprayed dense daily to field
It is 200 μm of methyl jasmonate solution of ol/L in Momordica grosvenori surface to spend, being dripped to surface, early, middle and late each sprinkling once,
Continuously spray 10d.
<Embodiment 6>
Step one, with volume fraction be 2% ethanol water dissolving methyl jasmonate, be configured to concentration for 200mmol/
The ethanol mother liquor of L methyl jasmonates, and with 0.22 μm of miillpore filter sterilizing, by the ethanol mother liquor of the methyl jasmonate after sterilizing
It is added in MS solid mediums, conditioning agent is added into MS solid mediums, the addition of conditioning agent is 5 μm of ol/L, jasmonic
The addition of methyl esters is 200 μm of ol/L, sterilizes and be cooled to 24~26 DEG C, and conditioning agent includes that mass ratio is 2:1:0.1 bamboo vinegar
Liquid, tuber of stemona water extract, Quercetin, MS solid mediums include：MS, 1.5mg/L 6- benzyls aminoadenine, 0.3mg/L indoles fourths
Acid, 3.5g/L agar, 30g/L sucrose and 1.0g/L activated carbons；
Step 2, Luohanguo With Plantlets of Tissue Culture is seeded in the MS solid mediums containing methyl jasmonate that step one is obtained
In, it is placed under the conditions of relative humidity 60~66%, 1300~1500Lux of intensity of illumination, light application time 8h/d, 21~25 DEG C of temperature
Culture 30d；Tissue culture process includes two benches, and first stage culture 15d changes a MS solid medium, is replaced every 3d
MS solid mediums be heated to being molten into liquefied mixture, sponge is immersed in liquefied mixture and is fully absorbed, then cover
Cover above the MS solid mediums changed, until next time removes and change when changing culture medium；Second stage culture 15d, often
A MS solid medium is changed every 5d, the 2d after changing every time is atomized to the MS solid mediums for being inoculated with tissue-cultured seedling
Treatment, Alevaire is casein solution that mass fraction is 1%；
Step 3, with volume fraction be 2% ethanol water dissolving methyl jasmonate, be configured to concentration for 200 μm of ol/
The methyl jasmonate solution of L, the tissue culture transplantation of seedlings after step one is cultivated to field, by the top ring of plant after Pollination of Luohanguo
Multiple annular incision are cut, two neighboring otch spacing 5mm first sprays flavonol solution, the Ran Houyong that mass fraction is 1% daily
Peach gum smears otch, finally uses cordate houttuynia bondage otch, and after Pollination of Luohanguo 25d, daily spray concentration is 200 μm of jasmines of ol/L
In Momordica grosvenori surface, being dripped to surface, early, middle and late each sprinkling once, continuously sprays 10d to jasmine acid methyl ester solution.
<Comparative example 1>
It is to carry out seed soaking in 200 μm of methyl jasmonate solution of ol/L that Momordica grosvenori seed is immersed in into concentration, then
Carry out conventional organization culture and obtain tissue-cultured seedling, tissue culture transplantation of seedlings to field is finally carried out into conventional field culture.
<Comparative example 2>
It is to be pre-processed in 200 μm of methyl jasmonate solution of ol/L that Momordica grosvenori explant is immersed in into concentration, then
Carry out conventional organization culture and obtain tissue-cultured seedling, tissue culture transplantation of seedlings to field is finally carried out into conventional field culture.
<Comparative example 3>
Luohanguo With Plantlets of Tissue Culture is seeded in MS solid mediums, relative humidity 60~66%, intensity of illumination 1300 is placed in
30d is cultivated under the conditions of~1500Lux, light application time 8h/d, 21~25 DEG C of temperature；Wherein, MS solid mediums include：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；
Then tissue culture transplantation of seedlings to field is carried out into conventional field culture.
<Momordica grosvenori quality test>
The sampling of the next day after methyl jasmonate solution is continuously sprayed to embodiment 2,5,6, to embodiment 1, comparative example 1~3
Fruit development mid-term pollination 30d post-samplings, count the damping-off incidence of disease and Momordica grosvenori shoot survival percent, while using LC-MS methods
Carry out Momordica grosvenori Simon glycosides I content detections.
Liquid phase chromatogram condition：Chromatographic column：Poroshell 120SB-C18(2.1mm×100mm,2.7μm)；Column temperature：30
℃；Mobile phase：0.1% aqueous formic acid (A)-acetonitrile (B), gradient elution (0-5min, 23%B；5-15min, 23%B are extremely
26%B；15-20min, 26%B to 33%B；20-25min, 23%B)；Flow velocity：0.3mL/min；The μ L of sample size 0.5.
Mass Spectrometry Conditions：Electric spray ion source (ESI)：Anion scan pattern；Selection ion monitoring mode (SIM)；Dry
Temperature degree：350℃；Dry gas stream speed：9L/min；Atomization gas pressure：176kPa；Capillary voltage：3.5kV.
The preparation of standard liquid：Momordica grosvenori Simon glycosides I 10mg (being accurate to 0.1mg) accurately is weighed, is dissolved with methyl alcohol and determined
Hold into 10mL volumetric flasks, be configured to 1000mg/L standard reserving solutions.4 DEG C of preservations of lucifuge.Accurately pipette appropriate above-mentioned standard
Storing solution, uses water constant volume, is configured to the standard working solution of series concentration, needs matching while using.
Sample treatment：After Momordica grosvenori sample comminution, 0.500g is accurately weighed in 50mL conical flasks, add ethanol 20mL,
Taken out after ultrasound (120W, 40kHz) 30min, mend weight.It is to be measured after 0.45 μm of membrane filtration.
The preparation of negative sample solution：Take 1mL ethanol direct injecteds.
Data are as shown in the table.
As seen from the above table, comparative example 1~2 is compared to comparative example 3, and the content of Simon glycosides I does not have significantly raised, illustrates leaching
Kind, explant be immersed in the content that methyl jasmonate solution is not obviously improved Momordica grosvenori Simon glycosides I, the content of embodiment 1,2
Compared to comparative example 3, the content of Simon glycosides I is increased significantly, and illustrates to induce plant by methyl jasmonate during tissue culture or cultivation
Strain growth, can be obviously improved the content of Simon glycosides I, and compared to comparative example 3, the content of Simon glycosides I has for the content of embodiment 5,6
Significantly raise, illustrate during tissue culture, cultivation by methyl jasmonate induction plant strain growth, Simon glycosides I can be obviously improved
Content.Additionally, embodiment 1,2,5,6, the damping-off incidence of disease of comparative example 1,2 are substantially less than comparative example 3, methyl jasmonate
The damping-off illness rate of Momordica grosvenori can also be significantly reduced, the Momordica grosvenori shoot survival percent of embodiment 1,2,5,6 is above comparative example 1
~3, Momordica grosvenori shoot survival percent can be improved by methyl jasmonate induction plant strain growth during illustrating tissue culture and/or cultivation.
Number of devices described herein and treatment scale are for simplifying explanation of the invention.To application of the invention,
Modifications and variations will be readily apparent to persons skilled in the art.
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 (9)
1. it is a kind of improve Momordica grosvenori in Simon glycosides I contents method, it is characterised in that in Momordica grosvenori tissue culture process or cultivation
Middle applying methyl jasmonate.
2. the method for improving Simon glycosides I contents in Momordica grosvenori as claimed in claim 1, it is characterised in that during tissue culture, will
Luohanguo With Plantlets of Tissue Culture is seeded in the MS solid mediums containing methyl jasmonate, is placed in relative humidity 60~66%, illumination strong
28~31d is cultivated under the conditions of 1300~1500Lux of degree, light application time 8h/d, 21~25 DEG C of temperature, wherein, methyl jasmonate
Addition is 50~400 μm of ol/L.
3. the method for improving Simon glycosides I contents in Momordica grosvenori as claimed in claim 1, it is characterised in that in cultivation, will
It is 50~400 μm of methyl jasmonate solution of ol/L that methyl jasmonate is made into concentration, after 20~30d of Pollination of Luohanguo, spray daily
Methyl jasmonate solution is spilt in Momordica grosvenori surface, being dripped to surface, early, middle and late each sprinkling once, continuously spray 8~
12d。
4. the method for improving Simon glycosides I contents in Momordica grosvenori as claimed in claim 1, it is characterised in that including：
Step one, Luohanguo With Plantlets of Tissue Culture is seeded in the MS solid mediums containing methyl jasmonate, is placed in relative humidity 60
~66%, 28~31d is cultivated under the conditions of 1300~1500Lux of intensity of illumination, light application time 8h/d, 21~25 DEG C of temperature, wherein,
The addition of methyl jasmonate is 50~400 μm of ol/L；
Step 2, step one is cultivated after tissue culture transplantation of seedlings to field, after 20~30d of Pollination of Luohanguo, daily spray concentration
It is 50~400 μm of methyl jasmonate solution of ol/L in Momordica grosvenori surface, being dripped to surface, early, middle and late each sprinkling one
It is secondary, continuously spray 8~12d.
5. as described in claim 2 or 4 improve Momordica grosvenori in Simon glycosides I contents method, it is characterised in that use volume fraction
Ethanol water for 2% dissolves methyl jasmonate, is configured to the ethanol mother liquor that concentration is 50mmol/L methyl jasmonates, is used in combination
0.22 μm of miillpore filter sterilizing, the ethanol mother liquor of the methyl jasmonate after sterilizing is added in MS solid mediums, to jasmine
Jasmine acid methyl acetate concentrations are 50~400 μm of ol/L, sterilize and be cooled to 24~26 DEG C, wherein, MS solid mediums include：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.
6. as described in claim 3 or 4 improve Momordica grosvenori in Simon glycosides I contents method, it is characterised in that use volume fraction
Ethanol water for 2% dissolves methyl jasmonate, and it is 50~400 μm of methyl jasmonate solution of ol/L to be configured to concentration.
7. the method for improving Simon glycosides I contents in Momordica grosvenori as claimed in claim 5, it is characterised in that MS solid mediums
Also include conditioning agent, the addition of conditioning agent is 5 μm of ol/L, and conditioning agent includes that mass ratio is 2:1:0.1 bamboo vinegar, tuber of stemona water
Extract, Quercetin.
8. the method for improving Simon glycosides I contents in Momordica grosvenori as claimed in claim 7, it is characterised in that tissue culture process includes
Two benches, first stage culture 15d, a MS solid medium is changed every 3~5d, the MS solid mediums heating being replaced
To liquefied mixture is molten into, sponge is immersed in liquefied mixture and is fully absorbed, be then covered by the MS solids training changed
Base top is supported, until next time removes and change when changing culture medium；Second stage culture 15d, changes a MS and consolidates every 5~7d
Body culture medium, the 2d after changing every time carries out atomization process to the MS solid mediums for being inoculated with tissue-cultured seedling, and Alevaire is matter
Amount fraction is 1% casein solution.
9. the method for improving Simon glycosides I contents in Momordica grosvenori as claimed in claim 6, it is characterised in that by plant after pollination
Top girdling multiple annular incision, two neighboring otch spacing 5mm, daily first sprinkling mass fraction be 1% flavonol it is molten
Liquid then with peach gum smear otch, finally use cordate houttuynia bondage otch, until start spray methyl jasmonate solution.
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