CN107360971A - Promote the method for Momordica grosvenori SS gene expressions - Google Patents
Promote the method for Momordica grosvenori SS gene expressions Download PDFInfo
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- CN107360971A CN107360971A CN201710629949.1A CN201710629949A CN107360971A CN 107360971 A CN107360971 A CN 107360971A CN 201710629949 A CN201710629949 A CN 201710629949A CN 107360971 A CN107360971 A CN 107360971A
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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
- 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
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Abstract
The invention discloses a kind of method for promoting Momordica grosvenori SS gene expressions, and Luohanguo With Plantlets of Tissue Culture is seeded in inducing culture of the methyl jasmonate concentration for 260 345 μm of ol/L and cultivates 28 30d.The methyl jasmonate that the present invention adds can induce the SS gene expressions in Luohanguo With Plantlets of Tissue Culture, and then promote the synthesis of momordica glycoside V.
Description
Technical field
The present invention relates to plant biological field.It is more particularly related to a kind of promote Momordica grosvenori SS gene expressions
Method.
Background technology
Momordica glycoside V is extracted in Guangxi special product economic plants --- and Momordica grosvenori, its sugariness are 300 times of sucrose, its heat
Amount is zero, has the effect of clearing heat and moistening lung antibechic, relax bowel, has preventive and therapeutic effect to obesity, constipation, diabetes etc., at present
Market is larger to the demand of momordica glycoside V, it is therefore desirable to improves plantation efficiency to improve the yield of momordica glycoside V with full
The needs in sufficient market.Momordica glycoside V belongs to cucurbitane type tetracyclic triterpenes material, and this seminar transcribes early stage according to Momordica grosvenori
Group data, have derived the possible biosynthesis pathways of sweet tea glycosides V.The precursor substance of momordica glycoside V biosynthesis is iso-amylene
Base pyrophosphoric acid (DMAPP) in diphosphonic acid (IPP) and 3,3- dimethyl alkene, the two is by mevalonic acid (MVA) and the red moss of methyl
Two approach of sugar alcohol phosphorylation (MEP) are formed, and MVA approach occurs in kytoplasm, and MEP approach occurs in plastid.From upper
The IPP or DMAPP for stating two approach form Mang ox base pyrophosphoric acid (GPP), IPP through Mang ox base pyrophosphate synthase (GPS) catalysis
With GPP under farnesyl pyrophosphate synthase (FPS) catalytic action so that formed farnesyl pyrophosphate (FPP), then through spiny dogfish
The catalysis of alkene synthase (SS) forms squalene, and further catalysis forms 2,3- oxidosqualenes to squalene epoxidase (SE), most
Afterwards in the presence of cucurbit dienol synthase (CS), CYP450 enzymes and glucosyltransferase, 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 SS genes is to momordica glycoside V
Biosynthesis plays decisive role.SS genes are overexpressed, the accumulation of momordica glycoside V can be promoted；If on the contrary, suppress SS
The expression of gene, the yield of momordica glycoside V will significantly reduce.It there are no in the prior art and promote Momordica grosvenori SS gene expressions
Report.
The content of the invention
It is an object of the invention to solve the problems, such as that the yield of momordica glycoside V is not high, and provide behind at least and will say
The advantages of bright.
In order to realize according to object of the present invention and further advantage, there is provided one kind promotes Momordica grosvenori SS gene expressions
Method, Luohanguo With Plantlets of Tissue Culture is seeded in methyl jasmonate concentration to cultivate 28- in 260-345 μm of ol/L inducing culture
30d。
Preferably, Luohanguo With Plantlets of Tissue Culture is placed in humidity 60-66%, intensity of illumination 1400lux, light application time 8h/d,
Cultivated under the conditions of 23 ± 2 DEG C of temperature.
Preferably, dissolve methyl jasmonate with the ethanol water that volume fraction is 2% in above-mentioned steps and be configured to jasmine
Jasmine acid methyl esters mother liquor, and being sterilized with miillpore filter, is cooled to 24-26 DEG C after sterilizing, then by the methyl jasmonate mother liquor after cooling
It is added in solid medium, is 260-345 μm of ol/L to methyl jasmonate ultimate density, obtains inducing culture.
Preferably, the solid medium component is：MS+6-BA 1.5mg/L+ indolebutyric acid 0.3mg/L+ agar
3.5g/L+ sucrose 30g/l+ activated carbons 1.0g/L.
Preferably, methyl jasmonate is dissolved with the ethanol water that volume fraction is 2%, methyl jasmonate is made into dense
Spend for 260-345 μm of ol/L methyl jasmonate solution, be sprayed on the Momordica grosvenori surface of 20-30d after pollination, be sprayed to sieve every time
Untill Chinese fruit surface is dripped, sprinkling daily three times, every 4-6h sprinklings once, continuously sprays 4-6d.
Preferably, the 20-30d after Momordica grosvenori seedling replanting, the regulation phosphorus solution that concentration is 300-500mg/L is used
Ultrasonic wave carries out processing 20-30min, supersonic frequency 25-35kHz, is wrapped up the top half of Momordica grosvenori seedling with black cotton
Firmly, while with cache-sex to Momordica grosvenori seedling shelter from heat or light, will persistently be dripped in black cotton by the regulation phosphorus solution being ultrasonically treated
On, duration 2-3h；Black cotton is removed, while removes cache-sex, the narcissus that mass concentration is 0.6-0.9% is taken after 6-8h
Plain solution, narcissus element solution is subjected to processing 20-25min, supersonic frequency 25-32kHz with ultrasonic wave, then with black cotton by sieve
The top half of Chinese fruit seedling is wrapped, while Momordica grosvenori seedling is sheltered from heat or light with cache-sex, by the water by being ultrasonically treated
Celestial plain solution is persistently dripped on black cotton, duration 1.5-2h, is removed black cotton afterwards, is removed cache-sex；It is right every 3 days
Momordica grosvenori seedling is handled once, coprocessing 10 times.
Preferably, flyash 25-30 parts by weight, urea 10-15 parts by weight, dead fish 20-25 parts by weight, Grape Skin 5- are taken
10 parts by weight, plant ash 10-15 parts by weight, rotten fruit juice 4-8 parts by weight and rare earth powder 1-3 parts by weight, by dead fish 20-25 weights
Measure part, Grape Skin 5-10 parts by weight and rotten fruit juice 4-8 parts by weight to fall in fermentation tank, add the water of 25-35 parts by weight
Fermented, after the completion of fermentation, taken out and flyash 25-30 parts by weight, urea 10-15 parts by weight, plant ash 10-15 parts by weight
And rare earth powder 1-3 parts by weight are well mixed, and mixed fertilizer is made, since the Momordica grosvenori plant florescence, the mixed fertilizer is embedded in sieve
Around Chinese fruit plant root, every plant is buried 0.2-0.5kg, then is watered by magnetized water, is afterwards 120-190mT with magnetic field intensity
Magnetic field the root of plant is handled, 5-10min per treatment, every 1h processing once, coprocessing is three times.
The present invention comprises at least following beneficial effect：
(1) jasmonic formicester is added in the culture medium of Luohanguo With Plantlets of Tissue Culture, so in Luohanguo With Plantlets of Tissue Culture growth course
In, jasmonic formicester acts on tissue-cultured seedling always, induces the SS gene expressions in Luohanguo With Plantlets of Tissue Culture, and then promote sweet Momordica grosvenori
Glycosides V synthesis, and the applicant is by a large amount of
(2) 20-30d Momordica grosvenori surface sprays jasmonic formicester solution after pollination, is used in Lo Han Guo fruit expanding stage
Jasmonic formicester is gone to stimulate Momordica grosvenori and then induces the expression of Momordica grosvenori SS genes, improves the content of momordica glycoside V；
(3) experiment finds that methyl jasmonate concentration is the SS in 260-345 μm of ol/L optimums induction Luohanguo With Plantlets of Tissue Culture
Gene expression；The molecule that phosphorus solution and narcissus element solution can be in solution is adjusted with ultrasonication to diminish easily by Momordica grosvenori
Plant absorbs, and regulation phosphorus can activate the rate-limiting enzyme of Momordica grosvenori, promote SS gene expressions, improve the yield of momordica glycoside V, use
Black cloth parcel carries out applying regulation phosphorus with the mode dripped again and narcissus element is more directly sprayed and is more easy to by Momordica grosvenori absorption and lasting work
Time is longer, and narcissus element can then induce Momordica grosvenori SS gene expressions, and it is to prevent from applying in drop that Momordica grosvenori, which shelter from heat or light,
When adjusting phosphorus and narcissus element, regulation phosphorus and narcissus element have been decomposed in illumination；
(4) present invention is first fermented with organic matters such as dead fish, Grape Skins, is advantageous to send out rich in sugar in Grape Skin and fruit juice
Ferment, the fertilizer after fermentation are rich in animal amino acid and holophytic nutrition, can promote the synthesis of momordica glycoside V, the fertilizer after fermentation
Material mixes with fertilizer, then is mixed with having magnetic flyash, the Momordica grosvenori root after imposing on the florescence as carrier using bentonite, makes
Obtaining Momordica grosvenori has enough nutrition in the phase of yielding positive results, and magnetization treatment is carried out to Momordica grosvenori after fertilising, and flyash can strengthen magnetic
Change acts on, and magnetization treatment can produce positive influences to the DNA content and nucleic acid of Momordica grosvenori, promote Momordica grosvenori SS genes
Expression, improves the content of momordica glycoside V, and the present inventor is found by experiment that 120-190mT magnetic field intensity to Momordica grosvenori
Action effect is optimal.
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 specific embodiment, the present invention is described in further detail, to make those skilled in the art's reference say
Bright book 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 for promoting Momordica grosvenori SS gene expressions, Luohanguo With Plantlets of Tissue Culture is seeded in into methyl jasmonate concentration is
In 260 μm of ol/L inducing culture, humidity 60%, intensity of illumination 1200lux, light application time 8h/d, 23 ± 2 DEG C of temperature are placed in
Under the conditions of cultivate 30d.Wherein solid medium component is：MS+6-BA 1.5mg/L+ indolebutyric acid 0.3mg/L+ agar 3.5g/L
+ sucrose 30g/l+ activated carbons 1.0g/L；Add methyl jasmonate method be：It is molten with the ethanol water that volume fraction is 2%
Solution methyl jasmonate is configured to methyl jasmonate mother liquor, and is sterilized with 0.22 μm of miillpore filter, and 24-26 is cooled to after sterilizing
DEG C, then the methyl jasmonate mother liquor after cooling is added in above-mentioned solid medium, it is 260 to methyl jasmonate ultimate density
μm ol/L, obtains inducing culture.
Embodiment 2：
A kind of method for promoting Momordica grosvenori SS gene expressions, Luohanguo With Plantlets of Tissue Culture is seeded in into methyl jasmonate concentration is
In 315 μm of ol/L inducing culture, humidity 65%, intensity of illumination 1400lux, light application time 8h/d, 23 ± 2 DEG C of temperature are placed in
Under the conditions of cultivate 30d.Wherein solid medium component is：MS+6-BA 1.5mg/L+ indolebutyric acid 0.3mg/L+ agar 3.5g/L
+ sucrose 30g/l+ activated carbons 1.0g/L；Add methyl jasmonate method be：It is molten with the ethanol water that volume fraction is 2%
Solution methyl jasmonate is configured to methyl jasmonate mother liquor, and is sterilized with 0.22 μm of miillpore filter, and 24-26 is cooled to after sterilizing
DEG C, then the methyl jasmonate mother liquor after cooling is added in above-mentioned solid medium, it is 315 to methyl jasmonate ultimate density
μm ol/L, obtains inducing culture.
Embodiment 3：
A kind of method for promoting Momordica grosvenori SS gene expressions, Luohanguo With Plantlets of Tissue Culture is seeded in into methyl jasmonate concentration is
In 345 μm of ol/L inducing culture, humidity 66%, intensity of illumination 1400lux, light application time 8h/d, 23 ± 2 DEG C of temperature are placed in
Under the conditions of cultivate 30d.Wherein solid medium component is：MS+6-BA 1.5mg/L+ indolebutyric acid 0.3mg/L+ agar 3.5g/L
+ sucrose 30g/l+ activated carbons 1.0g/L；Add methyl jasmonate method be：It is molten with the ethanol water that volume fraction is 2%
Solution methyl jasmonate is configured to methyl jasmonate mother liquor, and is sterilized with 0.22 μm of miillpore filter, and 24-26 is cooled to after sterilizing
DEG C, then the methyl jasmonate mother liquor after cooling is added in above-mentioned solid medium, it is 345 to methyl jasmonate ultimate density
μm ol/L, obtains inducing culture.
Embodiment 4：
On the basis of embodiment 2, methyl jasmonate is dissolved with the ethanol water that volume fraction is 2%, by jasmonic
Methyl esters is made into methyl jasmonate solution of the concentration for 265 μm of ol/L, is sprayed on the Momordica grosvenori surface of 30d after pollination, every time sprinkling
Untill Momordica grosvenori surface is dripped, sprinkling daily three times, every 4h sprinklings once, continuously sprays 6d.
Embodiment 5：
On the basis of embodiment 2, methyl jasmonate is dissolved with the ethanol water that volume fraction is 2%, by jasmonic
Methyl esters is made into methyl jasmonate solution of the concentration for 315 μm of ol/L, is sprayed on the Momordica grosvenori surface of 25d after pollination, every time sprinkling
Untill Momordica grosvenori surface is dripped, sprinkling daily three times, every 6h sprinklings once, continuously sprays 5d.
Embodiment 6：
On the basis of embodiment 2, methyl jasmonate is dissolved with the ethanol water that volume fraction is 2%, by jasmonic
Methyl esters is made into methyl jasmonate solution of the concentration for 345 μm of ol/L, is sprayed on the Momordica grosvenori surface of 20d after pollination, every time sprinkling
Untill Momordica grosvenori surface is dripped, sprinkling daily three times, every 5h sprinklings once, continuously sprays 4d.
Embodiment 7：
On the basis of embodiment 2, the 20d after Momordica grosvenori seedling replanting, the regulation phosphorus solution by concentration for 300mg/L,
Processing 20min, supersonic frequency 35kHz are carried out with ultrasonic wave, is wrapped the top half of Momordica grosvenori seedling with black cotton, together
When Momordica grosvenori seedling is sheltered from heat or light with cache-sex, will through supersound process regulation phosphorus solution persistently drip on black cotton, hold
The continuous time is 2h；Black cotton is removed, while removes cache-sex, the narcissus element solution that mass concentration is 0.9% is taken after 6h, by narcissus
Plain solution carries out processing 25min, supersonic frequency 25kHz with ultrasonic wave, then with black cotton by the top half of Momordica grosvenori seedling
Wrap, while Momordica grosvenori seedling is sheltered from heat or light with cache-sex, will persistently be dripped in black by the narcissus element solution being ultrasonically treated
On cotton, duration 1.5h, black cotton is removed afterwards, removes cache-sex；Processing one is carried out to Momordica grosvenori seedling every 3 days
It is secondary, coprocessing 10 times.
Embodiment 8：
On the basis of embodiment 2, the 25d after Momordica grosvenori seedling replanting, the regulation phosphorus solution by concentration for 410mg/L,
Processing 25min, supersonic frequency 32kHz are carried out with ultrasonic wave, is wrapped the top half of Momordica grosvenori seedling with black cotton, together
When Momordica grosvenori seedling is sheltered from heat or light with cache-sex, will through supersound process regulation phosphorus solution persistently drip on black cotton, hold
The continuous time is 3h；Black cotton is removed, while removes cache-sex, the narcissus element solution that mass concentration is 0.7% is taken after 7h, by narcissus
Plain solution carries out processing 20min, supersonic frequency 32kHz with ultrasonic wave, then with black cotton by the top half of Momordica grosvenori seedling
Wrap, while Momordica grosvenori seedling is sheltered from heat or light with cache-sex, will persistently be dripped in black by the narcissus element solution being ultrasonically treated
On cotton, duration 2h, black cotton is removed afterwards, removes cache-sex；Processing one is carried out to Momordica grosvenori seedling every 3 days
It is secondary, coprocessing 10 times.
Embodiment 9：
On the basis of embodiment 2, the 30d after Momordica grosvenori seedling replanting, the regulation phosphorus solution by concentration for 500mg/L,
Processing 30min, supersonic frequency 25kHz are carried out with ultrasonic wave, is wrapped the top half of Momordica grosvenori seedling with black cotton, together
When Momordica grosvenori seedling is sheltered from heat or light with cache-sex, will through supersound process regulation phosphorus solution persistently drip on black cotton, hold
The continuous time is 3h；Black cotton is removed, while removes cache-sex, the narcissus element solution that mass concentration is 0.6% is taken after 8h, by narcissus
Plain solution carries out processing 22min, supersonic frequency 28kHz with ultrasonic wave, then with black cotton by the top half of Momordica grosvenori seedling
Wrap, while Momordica grosvenori seedling is sheltered from heat or light with cache-sex, will persistently be dripped in black by the narcissus element solution being ultrasonically treated
On cotton, duration 2h, black cotton is removed afterwards, removes cache-sex；Processing one is carried out to Momordica grosvenori seedling every 3 days
It is secondary, coprocessing 10 times.
Embodiment 10：
On the basis of embodiment 9, the parts by weight of flyash 28, the parts by weight of urea 12, the parts by weight of dead fish 25, Grape Skin 5 are taken
Parts by weight, the parts by weight of plant ash 10, the rotten parts by weight of fruit juice 8 and the parts by weight of rare earth powder 1, by the parts by weight of dead fish 25, Grape Skin 5
Parts by weight and the parts by weight of rotten fruit juice 8 are fallen in fermentation tank, and the water for adding 32 parts by weight is fermented, after the completion of fermentation,
Taking-up is well mixed with the parts by weight of flyash 28, the parts by weight of urea 12, the parts by weight of plant ash 10 and the parts by weight of rare earth powder 1, is made
Into mixed fertilizer, since the Momordica grosvenori plant florescence, the mixed fertilizer is embedded in around Momordica grosvenori plant root, every plant is buried 0.25kg,
Water by magnetized water, the root of plant is handled with the magnetic field that magnetic field intensity is 125mT again afterwards, it is per treatment
10min, every 1h processing once, coprocessing is three times.
Embodiment 11：
On the basis of embodiment 9, the parts by weight of flyash 25, the parts by weight of urea 15, the parts by weight of dead fish 20, Grape Skin 8 are taken
Parts by weight, the parts by weight of plant ash 12, the rotten parts by weight of fruit juice 4 and the parts by weight of rare earth powder 2, by the parts by weight of dead fish 20, Grape Skin 8
Parts by weight and the parts by weight of rotten fruit juice 48 are fallen in fermentation tank, and the water for adding 26 parts by weight is fermented, after the completion of fermentation,
Taking-up is well mixed with the parts by weight of flyash 25, the parts by weight of urea 15, the parts by weight of plant ash 12 and the parts by weight of rare earth powder 2, is made
Into mixed fertilizer, since the Momordica grosvenori plant florescence, the mixed fertilizer is embedded in around Momordica grosvenori plant root, every plant is buried 0.38kg,
Water by magnetized water, the root of plant is handled with the magnetic field that magnetic field intensity is 190mT again afterwards, it is per treatment
5min, every 1h processing once, coprocessing is three times.
Embodiment 12：
On the basis of embodiment 9, the parts by weight of flyash 30, the parts by weight of urea 10, the parts by weight of dead fish 23, Grape Skin 10 are taken
Parts by weight, the parts by weight of plant ash 15, the rotten parts by weight of fruit juice 6 and the parts by weight of rare earth powder 3, by the parts by weight of dead fish 23, Grape Skin
10 parts by weight and the parts by weight of rotten fruit juice 6 are fallen in fermentation tank, and the water for adding 35 parts by weight is fermented, and fermentation is completed
Afterwards, take out and be well mixed with the parts by weight of flyash 30, the parts by weight of urea 10, the parts by weight of plant ash 15 and the parts by weight of rare earth powder 3,
Mixed fertilizer is made, since the Momordica grosvenori plant florescence, the mixed fertilizer is embedded in around Momordica grosvenori plant root, every plant is buried 0.5kg,
Water by magnetized water, the root of plant is handled with the magnetic field that magnetic field intensity is 170mT again afterwards, it is per treatment
8min, every 1h processing once, coprocessing is three times.
In order to illustrate beneficial effects of the present invention, the arhat that applicant of the present invention obtains for distinct methods or embodiment
Fruits, it is measured for SS gene expression amounts and sweet tea glycosides V content, specific method and result are as follows：
1st, SS gene expressions quantity measuring method：Using ABI7500 real-time fluorescence quantitative PCR instrument, SS is detected using qRT-PCR
The expression of gene.
Step 1: sample pre-treatments：Lo Han Guo fruit is gathered, picking pulp, 2-4mm fritters is cut into and uses masking foil respectively
Wrap, be immediately placed on it is quick-frozen in liquid nitrogen, be stored in -80 DEG C it is standby.
Step 2: first using improved Trizol method extraction Lo Han Guo fruit total serum IgE；
Step 3: it is μ L, the PrimeScript RT Enzyme of RNA 10.0 by the reaction system that RNA reverse transcriptions are cDNA
μ L, the RNase Free dH2O of 1.0 μ L, RT Primer Mix of Mix I, 1.0 μ L, 5 × PrimeScript Buffer 2 4.0
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, had by raw work bioengineering (Shanghai) share
Limit company synthesizes, wherein, SS primer sequences are：
FP：CTGAGACACCCAGATGACT, RP：GAGGGCTCGCAGAACAAGA；
Reference gene UBQ5, sequence are：
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
The 2.0 6.0 μ L of μ L, dH2O of μ L, Template of Reference Dye of Dye II (50 ×) 0.4.Reaction condition is 95 DEG C
(30s), then carry out 40 cycles [95 DEG C (5s), 95 DEG C (34s)].
2nd, momordica glycoside V content：Using high performance liquid chromatography, specific method reference《Colleges Of Traditional Chinese Medicine Of Guangxi's journal》The
" content of momordica glycoside V in the HPLC measure Momordica grosvenoris " text delivered on 04 phase in 2007.
Momordica grosvenori, routine side are planted with the method for conventional method, embodiment 2, embodiment 5, embodiment 9 and embodiment 12
Method is planted as a control group, is won 30 pieces of Momordica grosvenoris altogether on the different Momordica grosvenori plants of every kind of method plantation, is carried out experiment and ask
The SS gene expression amounts and momordica glycoside V content being averagely worth to are as shown in table 1：
Table 1
As can be seen from Table 1, the Momordica grosvenori SS gene expression amounts of embodiment 2 and momordica glycoside V content are apparently higher than right
According to group, it can be seen that increase jasmonic formicester in the solid medium in Momordica grosvenori tissue culture face to improving Momordica grosvenori SS gene expressions
Amount and momordica glycoside V content are significantly effective.
Embodiment 5 is that jasmonic formicester solution is sprayed on the Momordica grosvenori on the basis of embodiment 2 after pollination, from table 1
In as can be seen that the Momordica grosvenori SS gene expression amounts and momordica glycoside V content of embodiment 5 apparently higher than embodiment 2, therefore can
Jasmonic formicester solution is sprayed to Momordica grosvenori after pollination with explanation also can effectively induce Momordica grosvenori SS gene expressions, Jin Erti
High momordica glycoside V content.
Embodiment 9 is on the basis of embodiment 2, Momordica grosvenori is sprayed phosphorus solution and narcissus are adjusted after supersound process
Plain solution, from table 1 it follows that the Momordica grosvenori SS gene expression amounts of embodiment 9 and momordica glycoside V content are apparently higher than reality
Example 2 is applied, is diminished easily quilt because adjusting the molecule that phosphorus solution and narcissus element solution can be in solution with ultrasonication
Momordica grosvenori plant absorbs, and regulation phosphorus can activate the rate-limiting enzyme of Momordica grosvenori.
Embodiment 12 is on the basis of embodiment 9, and homemade mixed fertilizer is applied to Momordica grosvenori plant and is carried out after fertilising
Magnetization treatment, from table 1 it follows that the Momordica grosvenori SS gene expression amounts of embodiment 12 and momordica glycoside V content are substantially high
In embodiment 9, magnetization treatment can produce positive influences to the DNA content and nucleic acid of Momordica grosvenori, can induce arhat
The expression of fruit SS genes, and then improve the content of momordica glycoside V.
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, it is of the invention and unlimited
In specific details and shown here as the embodiment with description.
Claims (7)
- A kind of 1. method for promoting Momordica grosvenori SS gene expressions, it is characterised in that Luohanguo With Plantlets of Tissue Culture is seeded in jasmonic first Ester concentration is to cultivate 28-30d in 260-345 μm of ol/L inducing culture.
- 2. promote the method for Momordica grosvenori SS gene expressions as claimed in claim 1, it is characterised in that put Luohanguo With Plantlets of Tissue Culture In humidity 60-66%, intensity of illumination 1400lux, light application time 8h/d, cultivate under the conditions of 23 ± 2 DEG C of temperature.
- 3. promote the method for Momordica grosvenori SS gene expressions as claimed in claim 1, it is characterised in that volume is used in above-mentioned steps The ethanol water that fraction is 2% dissolves methyl jasmonate and is configured to methyl jasmonate mother liquor, and is sterilized with miillpore filter, sterilizes After be cooled to 24-26 DEG C, then the methyl jasmonate mother liquor after cooling is added in solid medium, it is final to methyl jasmonate Concentration is 260-345 μm of ol/L, obtains inducing culture.
- 4. promote the method for Momordica grosvenori SS gene expressions as claimed in claim 3, it is characterised in that the solid medium group It is divided into：MS+6-BA 1.5mg/L+ indolebutyric acid 0.3mg/L+ agar 3.5g/L+ sucrose 30g/l+ activated carbons 1.0g/L.
- 5. as claimed in claim 1 promote Momordica grosvenori SS gene expressions method, it is characterised in that with volume fraction be 2% Ethanol water dissolving methyl jasmonate, by methyl jasmonate be made into concentration be 260-345 μm of ol/L methyl jasmonate it is molten Liquid, be sprayed on the Momordica grosvenori surface of 20-30d after pollination, be sprayed to every time untill Momordica grosvenori surface drips, sprinkling daily three times, Every 4-6h sprinklings once, 4-6d is continuously sprayed.
- 6. promote the method for Momordica grosvenori SS gene expressions as claimed in claim 1, it is characterised in that in Momordica grosvenori seedling replanting 20-30d afterwards, by the regulation phosphorus solution that concentration is 300-500mg/L, processing 20-30min is carried out with ultrasonic wave, supersonic frequency is 25-35kHz, the top half of Momordica grosvenori seedling is wrapped with black cotton, while Momordica grosvenori seedling hidden with cache-sex The moon, will persistently it be dripped on black cotton by the regulation phosphorus solution being ultrasonically treated, duration 2-3h；Black cotton is removed, simultaneously Remove cache-sex, the narcissus element solution that mass concentration is 0.6-0.9% is taken after 6-8h, by narcissus element solution with ultrasonic wave 20-25min, supersonic frequency 25-32kHz are managed, then is wrapped the top half of Momordica grosvenori seedling with black cotton, while with screening Cloudy cloth shelters from heat or light to Momordica grosvenori seedling, will persistently be dripped on black cotton by the narcissus element solution being ultrasonically treated, the duration For 1.5-2h, black cotton is removed afterwards, removes cache-sex；Momordica grosvenori seedling is handled once every 3 days, coprocessing 10 It is secondary.
- 7. promote the method for Momordica grosvenori SS gene expressions as claimed in claim 1, it is characterised in that take flyash 25-30 weights Measure part, urea 10-15 parts by weight, dead fish 20-25 parts by weight, Grape Skin 5-10 parts by weight, plant ash 10-15 parts by weight, rotten Fruit juice 4-8 parts by weight and rare earth powder 1-3 parts by weight, by dead fish 20-25 parts by weight, Grape Skin 5-10 parts by weight and rotten fruit Juice 4-8 parts by weight are fallen in fermentation tank, and the water for adding 25-35 parts by weight is fermented, after the completion of fermentation, taking-up and flyash 25-30 parts by weight, urea 10-15 parts by weight, plant ash 10-15 parts by weight and rare earth powder 1-3 parts by weight are well mixed, and are made Mixed fertilizer, since the Momordica grosvenori plant florescence, the mixed fertilizer is embedded in around Momordica grosvenori plant root, every plant is buried 0.2-0.5kg, Water by magnetized water, the root of plant is handled with the magnetic field that magnetic field intensity is 120-190mT again afterwards, every time place 5-10min is managed, every 1h processing once, coprocessing is three times.
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