CN107691127A - A kind of radix tylophorae root media and preparation method - Google Patents
A kind of radix tylophorae root media and preparation method Download PDFInfo
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- CN107691127A CN107691127A CN201711037125.1A CN201711037125A CN107691127A CN 107691127 A CN107691127 A CN 107691127A CN 201711037125 A CN201711037125 A CN 201711037125A CN 107691127 A CN107691127 A CN 107691127A
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- radix tylophorae
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Abstract
The present invention relates to seed processing technology field, and in particular to a kind of radix tylophorae root media and preparation method.Radix tylophorae root media of the present invention is added in White improved culture mediums：The 7g/L of agar 3, the 30mg/L of sweet potato mash 20, the 18mg/L of jasmonic 10, fast 1 4mg/L of 6-benzylamino, the 6mg/L of indolebutyric acid 2, the 4mg/L of ethephon (CEPHA),2-(chloroethyl) phosphonic acid 1, the 9mg/L of rape element sterol 4, the 12mg/L of lactoalbumin hydrolysate 4, the 16mg/L of copper coin grass liquid 8, the 14mg/L of Dragonfruit Juice 6, vitamin B6 are 0.4 1.2mg/L.The present invention provides a kind of radix tylophorae root media, and the culture medium can promote radix tylophorae tissue-cultured seedling rooting rate and quantity, improve tissue-cultured seedling adaptability, improve survival rate after tissue culture transplantation of seedlings；Another object of the present invention, there is provided the preparation method of the culture medium.
Description
【Technical field】
The present invention relates to botanical seedling culturing culture medium technical field, and in particular to a kind of radix tylophorae root media and preparation side
Method.
【Background technology】
Radix tylophorae is the perennial grape rattan shape grass of Asclepiadaceae (Asclepiadaceac) Tylophora (Tylophora)
This, mainly with root and all herbal medicine, mainly respectively at Yunnan Province of China, Guangxi, Guangdong, Hunan, Jiangxi, Zhejiang, Taiwan, Vietnam,
The ground such as Laos, Burma, India, happiness is warm, can shade tolerant, it is relatively cold-resistant, be frequently grown in Shrubs of Mountainous Areas of the height above sea level below 900, or
In mountain valley and on the sunny side density spinney.The climing bar of radix tylophorae curls up, and plants suitable for wattled wall railing, door frame, flower corridor, can also intersperse in vacation
Between mountain and rock slope slot, climb around thereon, climing bar is sagging, rather unique.
Radix tylophorae, which is used as medicine, long history in China,《Jiangxi herbal medicine》Record it there is dispelling pathogenic wind and eliminating phlegm, scattered addiction of stimulating the menstrual flow
Effect.《Nanning City's medicine will》Recording it has promoting the circulation of qi, dissipates addiction, analgesic, resolving sputum, cough-relieving, controls bruise, knife wound, cough, rheumatalgia
Effect.Jiangxi《Herbal medicine handbook》Record its can dissipate addiction, it is emetic control asthma productive cough, abscess of throat, traumatic injury, rheumatalgia, stomachache,
Stomachache etc..Modern pharmacological research shows, radix tylophorae primary bioactive components are alkaloid compounds, people's passing through
Learn a series of tylophorine derivatives of the synthetic method synthesis including tylophorine, and the derivative of these tylophorines
With the obvious pharmacological activity of antitumor, anti-inflammatory, antiviral, anticancer etc..
Radix tylophorae is used for town pain caused by cancer Guangxi is among the people, and effective in cure good, due to excavating year after year, wild resource spontaneous pollination rate
Low, conventional growth environment is destroyed, and radix tylophorae price of medicinal material remains high at present, and is in Critical Condition.Using tissue
Culture technique series radix tylophorae turns into one of important stock breeding approach, less to the Study on tissue culture of radix tylophorae at present,
Lack a kind of culture medium for effectively facilitating radix tylophorae tissue culture seedling rooting.
【The content of the invention】
The goal of the invention of the present invention is：For technical problem in above-mentioned background technology be present, the present invention provides a kind of baby
Youngster's rattan root media, the culture medium can promote radix tylophorae tissue-cultured seedling rooting rate and quantity, improve tissue-cultured seedling adaptability, carry
Survival rate after high tissue culture transplantation of seedlings；Another object of the present invention, there is provided the preparation method of the culture medium.
To achieve these goals, the technical solution adopted by the present invention is as follows：
A kind of radix tylophorae root media, add in White improved culture mediums：Agar, jasmonic, sweet potato mash, 6-benzyl
Amino is fast, indolebutyric acid, ethephon (CEPHA),2-(chloroethyl) phosphonic acid, rape element sterol, lactoalbumin hydrolysate, copper coin grass liquid, Dragonfruit Juice and vitamin B6.
As preferable, added in White improved culture mediums：Agar 3-7g/L, sweet potato mash 20-30mg/L, jasmonic 10-
The fast 1-4mg/L of 18mg/L, 6-benzylamino, indolebutyric acid 2-6mg/L, ethephon (CEPHA),2-(chloroethyl) phosphonic acid 1-4mg/L, rape element sterol 4-9mg/L,
Lactoalbumin hydrolysate 4-12mg/L, copper coin grass liquid 8-16mg/L, Dragonfruit Juice 6-14mg/L, vitamin B6 0.4-1.2mg/L.
It is preferable as another kind, added in White improved culture mediums：Agar 5g/L, sweet potato mash 25mg/L, jasmonic
The fast 2.5mg/L of 14mg/L, 6-benzylamino, indolebutyric acid 4mg/L, ethephon (CEPHA),2-(chloroethyl) phosphonic acid 2.5mg/L, rape element sterol 6.5mg/L, water
Solve lactoprotein 8mg/L, copper coin grass liquid 12mg/L, Dragonfruit Juice 10mg/L, vitamin B6 0.8mg/L.
Preferable as another kind, the preparation method of the copper coin grass liquid is：Copper coin grass is taken, after clean root is washed with water,
The lower root of shearing, after 85% alcohol-pickled 10-15min, takes out, adds the sterilized water of equivalent weight, smash stirring, filter paper mistake
Filter and produce.
Preferable as another kind, the preparation method of the Dragonfruit Juice is：Get fire imperial pulp and seed, be put into high-speed stirred
Stir, taken out with 400 mesh filter-cloth filterings in machine, then with centrifuge 5-10min, take upper strata to produce Dragonfruit Juice.
Preferable as another kind, the pH value of the radix tylophorae root media is 5.7-5.9.
It is preferable as another kind, contain a great number of elements in the improvement White culture mediums：Potassium nitrate 48mg/L, chlorine ester
Potassium 39mg/L, sodium dihydrogen phosphate 11.5mg/L, magnesium sulfate 442.8mg/L, calcium nitrate 172mg/L；Trace element：KI
0.5mg/L, boric acid 3.72mg/L, manganese nitrate 13.4mg/L, zinc sulfate 5.16mg/L, sodium molybdate 0.15mg/L, copper sulphate
0.015mg/L, cobalt chloride 0.015mg/L；Molysite：Disodium ethylene diamine tetraacetate 22.38mg/L, ferrous sulfate 16.68mg/L；Have
Machine composition：Inositol 80mg/L, glycine 1.2mg/L, thiamine hydrochloride 0.06mg/L, citric acid 1.2mg/L, puridoxine hydrochloride
0.3mg/L, nicotinic acid 0.3mg/L, sucrose 30mg/L, agar 7mg/L.
A kind of method of radix tylophorae root media, comprises the following steps：By weight, take sweet potato mash 1/3-1/2 total
The dissolving stirring of water water, then adds fast jasmonic, 6-benzylamino, indolebutyric acid, ethephon (CEPHA),2-(chloroethyl) phosphonic acid, oil in the case of stirring
Dish element sterol, lactoalbumin hydrolysate, copper coin grass liquid, Dragonfruit Juice, vitamin B6, continue to stir 20-25 minutes, obtain solution I；Then
Improvement White culture medium prescriptions are taken, are dissolved with 1/3-1/2 total water consumptions, then are well mixed with solution I, agar is added, is determined with water
Hold to required scale, regulation pH value continues to stir 15-20min between 5.7-5.9, filling, and culture medium then is put into 120-
126 DEG C, sterilize under conditions of 0.22-0.26MPa 15-25min, taking-up is let cool, and produces radix tylophorae root media.
In summary, by adopting the above-described technical solution, the beneficial effects of the invention are as follows：
The speed grown under radix tylophorae natural conditions is slow, and rate of yielding positive results is low, and the weaker cause of ability of environment is adapted to after transplanting
Make the cultivating and growing of radix tylophorae less, the speed and tissue-cultured seedling grown when laboratory cultures carries out hardening or transplanting to seedling
Adaptability is weak during garden, and not high survival rate is always that the kind can not introduce a fine variety the reason for plantation on a large scale.
The present invention is directly connected to radix tylophorae after propagation seedling transplanting for the root media during radix tylophorae tissue culture
Rooting rate, quantity of taking root and tissue-cultured seedling characteristic, according to the growth characteristic of radix tylophorae, by constantly studying, at a high speed, finally sieve
Select the culture medium of suitable radix tylophorae culture of rootage.By study find White culture mediums relative to conventional MS culture mediums and
B6 culture mediums, it can more meet the culture of rootage of radix tylophorae；But undesirable effect, therefore this hair are also only reached with this culture medium
It is bright further with the addition of in basal medium promote the release of radix tylophorae auxin, 6-benzylamino for stimulating vitality is fast, indoles
Butyric acid, ethephon (CEPHA),2-(chloroethyl) phosphonic acid, rape element sterol, vitamin B6 etc., these materials cooperate, and can inspire in radix tylophorae tissue-cultured seedling and stop
Dormancy material, suppress apical dominance, hestening rooting；Coordinate lactoalbumin hydrolysate, copper coin grass liquid, Dragonfruit Juice, it is common that abundant battalion is provided
While supporting, quantity of taking root can be reached at 7 days up to more than 5, the diameter of root is thicker, up to more than 1.5mm；Copper coin grass liquid and fire
Long Guozhong contains the nutriment for being especially suitable for root merisis, improving root adaptability in the past, add after both compositions with
It is not added with comparing, the survival rate after tissue culture transplantation of seedlings can be improved, reaches 87%, and the survival rate of un-added tissue culture transplantation of seedlings
Less than 72%.
【Embodiment】
With reference to embodiment, the invention will be further described.
Embodiment 1
The preparation method of copper coin grass liquid is：Copper coin grass is taken, after clean root is washed with water, lower root is sheared, with 85% alcohol
After soaking 10-15min, take out, add the sterilized water of equivalent weight, smash stirring, filter paper, which filters, to be produced.
It is appreciated that 85% alcohol-pickled time can also be to appoint in 10min, 15min or 10-15min in the present embodiment
One numerical value, person can realize the object of the invention.
Embodiment 2
The preparation method of Dragonfruit Juice is：Get fire imperial pulp and seed, be put into homogenizer and stir, take out with 400 mesh
Filter-cloth filtering, then with centrifuge 5-10min, take upper strata to produce Dragonfruit Juice.
It is appreciated that any value, person can realize in centrifuge 5min, 10min or 5-10min in the present embodiment
The object of the invention.
Embodiment 3
A kind of method of radix tylophorae root media, comprises the following steps：By weight, sweet potato mash is taken with 1/3 total Water
Water dissolving stirring, fast jasmonic, 6-benzylamino, indolebutyric acid, ethephon (CEPHA),2-(chloroethyl) phosphonic acid, rape element are then added in the case of stirring
Sterol, lactoalbumin hydrolysate, copper coin grass liquid, Dragonfruit Juice, vitamin B6, continue stirring 20 minutes, obtain solution I；Then improvement is taken
White culture medium prescriptions, are dissolved with 1/3 total water consumption, then are well mixed with solution I, add agar, required quarter is settled to water
Degree, regulation pH value continues to stir 15min between 5.7, filling, and culture medium then is put into 120 DEG C, under conditions of 0.22MPa
Sterilized 25min, and taking-up is let cool, and produces radix tylophorae root media.
Embodiment 4
A kind of method of radix tylophorae root media, comprises the following steps：By weight, sweet potato mash is taken with 1/2 total Water
Water dissolving stirring, fast jasmonic, 6-benzylamino, indolebutyric acid, ethephon (CEPHA),2-(chloroethyl) phosphonic acid, rape element are then added in the case of stirring
Sterol, lactoalbumin hydrolysate, copper coin grass liquid, Dragonfruit Juice, vitamin B6, continue to stir 20-25 minutes, obtain solution I；Then take and change
Good White culture medium prescriptions, are dissolved with 1/2 total water consumption, then are well mixed with solution I, add agar, needed for water is settled to
Scale, regulation pH value continues to stir 20min between 5.9, filling, and culture medium then is put into 126 DEG C, 0.26MPa condition
Lower sterilizing 15min, taking-up are let cool, and produce radix tylophorae root media.
Embodiment 5
A kind of method of radix tylophorae root media, comprises the following steps：By weight, sweet potato mash is taken with 1/3 total Water
Water dissolving stirring, fast jasmonic, 6-benzylamino, indolebutyric acid, ethephon (CEPHA),2-(chloroethyl) phosphonic acid, rape element are then added in the case of stirring
Sterol, lactoalbumin hydrolysate, copper coin grass liquid, Dragonfruit Juice, vitamin B6, continue stirring 22 minutes, obtain solution I；Then improvement is taken
White culture medium prescriptions, are dissolved with 1/3 total water consumption, then are well mixed with solution I, add agar, required quarter is settled to water
Degree, regulation pH value continues to stir 18min between 5.8, filling, and culture medium then is put into 123 DEG C, under conditions of 0.25MPa
Sterilized 20min, and taking-up is let cool, and produces radix tylophorae root media.
The root media of embodiment 6 is studied radix tylophorae tissue-cultured seedling rooting efficiency
Guangxi radix tylophorae stem seedling is chosen as explant, Multiple Buds is obtained after Fiber differentiation, Multiplying culture, will grow
The Multiple Buds of 2.5cm height are inoculated on laboratory sample 1-3 and comparative sample 1-2 of the present invention root media.Wherein test
Sample is formulated by laboratory sample 1-3 in table 1, is then prepared by embodiment 1,2 and 5 methods；Comparative example 1-2
It is formulated by the comparative sample 1-2 of formula table 1, cultivates referring next to embodiment 5 or routinely bitter cultural method and be prepared；
The selection of culture environment：After Multiple Buds are inoculated on root media, it is 2000lx to be placed in intensity of illumination, in light
It is that 25 DEG C of dark temperature are 20 DEG C according to temperature, culturing room's air humidity is 40%, under conditions of hour/day of light application time 8, culture
20 days, record quantity of taking root；Then take 15 rooted seedlings to remove culture medium in each sample to transplant to nursery, after planting 7 days, see
Examine seedling survival condition.Each sample equality is tested 3 times, is averaged, experimental result is shown in Table 2.
The each sample prescription of rooting medium situation table of table 1
The formula for improveing White culture mediums is as follows：A great number of elements：Potassium nitrate 48mg/L, chlorine ester potassium 39mg/L, biphosphate
Sodium 11.5mg/L, magnesium sulfate 442.8mg/L, calcium nitrate 172mg/L；Trace element：KI 0.5mg/L, boric acid 3.72mg/L,
Manganese nitrate 13.4mg/L, zinc sulfate 5.16mg/L, sodium molybdate 0.15mg/L, copper sulphate 0.015mg/L, cobalt chloride 0.015mg/L；
Molysite：Disodium ethylene diamine tetraacetate 22.38mg/L, ferrous sulfate 16.68mg/L；Organic principle：Inositol 80mg/L, glycine
1.2mg/L, thiamine hydrochloride 0.06mg/L, citric acid 1.2mg/L, puridoxine hydrochloride 0.3mg/L, nicotinic acid 0.3mg/L, sucrose
30mg/L, agar 7mg/L.
The each sample experimental result of table 2
| Sample 1 | Sample 2 | Sample 3 | Comparative sample 1 | Comparative sample 2 | |
| Inoculation material number (individual) | 30 | 30 | 30 | 30 | 30 |
| The material number (individual) taken root | 26 | 25 | 28 | 21 | 23 |
| Rooting rate (%) | 86.7 | 83.3 | 93.3 | 70 | 76.7 |
| Average every seedling rooting number (bar) | 8.1 | 7.6 | 9.8 | 5.2 | 6.4 |
| Transplant number (individual) | 15 | 15 | 15 | 15 | 15 |
| Transplant survival number (individual) | 13 | 12 | 13 | 8 | 10 |
| Transplant survival rate % | 86.7 | 80 | 86.7 | 53.3 | 66.7 |
As can be known from the above table, sample 1-3 is compared with comparative sample, rooting rate and transplanting survival rate apparently higher than comparative sample,
It is more suitable for radix tylophorae to take root, can also improves it and transplant survival rate.
Described above is the detailed description for the present invention preferably possible embodiments, but embodiment is not limited to this hair
Bright patent claim, the equal change completed or modification change under the technical spirit suggested by all present invention, all should belong to
Cover the scope of the claims in the present invention.
Claims (8)
1. a kind of radix tylophorae root media, it is characterised in that added in White improved culture mediums：Agar, jasmonic, sweet potato
Mud, 6-benzylamino are fast, indolebutyric acid, ethephon (CEPHA),2-(chloroethyl) phosphonic acid, rape element sterol, lactoalbumin hydrolysate, copper coin grass liquid, Dragonfruit Juice and dimension
Raw plain B6.
2. a kind of radix tylophorae root media according to claim 1, it is characterised in that add in White improved culture mediums
Enter：Agar 3-7g/L, sweet potato mash 20-30mg/L, jasmonic 10-18mg/L, the fast 1-4mg/L of 6-benzylamino, indolebutyric acid 2-
6mg/L, ethephon (CEPHA),2-(chloroethyl) phosphonic acid 1-4mg/L, rape element sterol 4-9mg/L, lactoalbumin hydrolysate 4-12mg/L, copper coin grass liquid 8-16mg/L, fire
Imperial fruit juice 6-14mg/L, vitamin B6 0.4-1.2mg/L.
3. a kind of radix tylophorae root media according to claim 2, it is characterised in that add in White improved culture mediums
Enter：Agar 5g/L, sweet potato mash 25mg/L, jasmonic 14mg/L, the fast 2.5mg/L of 6-benzylamino, indolebutyric acid 4mg/L, ethene
Sharp 2.5mg/L, rape element sterol 6.5mg/L, lactoalbumin hydrolysate 8mg/L, copper coin grass liquid 12mg/L, Dragonfruit Juice 10mg/L, dimension
Raw plain B6 is 0.8mg/L.
A kind of 4. radix tylophorae root media according to claim 1, it is characterised in that the preparation side of the copper coin grass liquid
Method is：Copper coin grass is taken, after clean root is washed with water, lower root is sheared, after 85% alcohol-pickled 10-15min, takes out, add
The sterilized water of equivalent weight, stirring is smashed, filter paper, which filters, to be produced.
A kind of 5. radix tylophorae root media according to claim 1, it is characterised in that the preparation side of the Dragonfruit Juice
Method is：Get fire imperial pulp and seed, be put into homogenizer and stir, take out with 400 mesh filter-cloth filterings, then with centrifuge 5-
10min, upper strata is taken to produce Dragonfruit Juice.
A kind of 6. radix tylophorae root media according to claim 2, it is characterised in that the radix tylophorae root media
PH value be 5.7-5.9.
A kind of 7. radix tylophorae root media according to claim 1, it is characterised in that the improvement White culture mediums
In contain a great number of elements：Potassium nitrate 48mg/L, chlorine ester potassium 39mg/L, sodium dihydrogen phosphate 11.5mg/L, magnesium sulfate 442.8mg/L,
Calcium nitrate 172mg/L；Trace element：KI 0.5mg/L, boric acid 3.72mg/L, manganese nitrate 13.4mg/L, zinc sulfate
5.16mg/L, sodium molybdate 0.15mg/L, copper sulphate 0.015mg/L, cobalt chloride 0.015mg/L；Molysite：Disodium ethylene diamine tetraacetate
22.38mg/L, ferrous sulfate 16.68mg/L；Organic principle：Inositol 80mg/L, glycine 1.2mg/L, thiamine hydrochloride
0.06mg/L, citric acid 1.2mg/L, puridoxine hydrochloride 0.3mg/L, nicotinic acid 0.3mg/L, sucrose 30mg/L, agar 7mg/L.
A kind of 8. method for preparing any one of the claim 2-7 radix tylophorae root medias, it is characterised in that including following
Step：By weight, take sweet potato mash 1/3-1/2 total Waters water dissolving stirring, then in the case of stirring add jasmonic,
6-benzylamino is fast, indolebutyric acid, ethephon (CEPHA),2-(chloroethyl) phosphonic acid, rape element sterol, lactoalbumin hydrolysate, copper coin grass liquid, Dragonfruit Juice, vitamin
B6, continue to stir 20-25 minutes, obtain solution I；Then improvement White culture medium prescriptions are taken, are dissolved with 1/3-1/2 total water consumptions,
It is well mixed again with solution I, adds agar, is settled to required scale with water, regulation pH value continues to stir between 5.7-5.9
15-20min, it is filling, culture medium is then put into 120-126 DEG C, sterilize under conditions of 0.22-0.26MPa 15-25min, taken out
Let cool, produce radix tylophorae root media.
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| CN115024326A (en) * | 2022-08-12 | 2022-09-09 | 山东爱普生农业发展有限公司 | Slow-release plant growth regulator and preparation method and application thereof |
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