CN109618933A - A kind of radix pseudostellariae Seed embryo culture base and its cultural method - Google Patents
A kind of radix pseudostellariae Seed embryo culture base and its cultural method Download PDFInfo
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- CN109618933A CN109618933A CN201910062789.6A CN201910062789A CN109618933A CN 109618933 A CN109618933 A CN 109618933A CN 201910062789 A CN201910062789 A CN 201910062789A CN 109618933 A CN109618933 A CN 109618933A
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- radix pseudostellariae
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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 radix pseudostellariae Seed embryo culture base and its cultural methods.The radix pseudostellariae Seed embryo culture base are as follows: contain sucrose 28-32g, agar powder 7.2-7.8g, 6- benzyl aminoadenine 0.40-0.60mg, methyl α-naphthyl acetate 0.10-0.30mg and heteroauxin 0.15-0.25mg in every liter of MS culture medium；The pH value of the radix pseudostellariae Seed embryo culture base is 5.6-6.0；The cultural method of the radix pseudostellariae Seed embryo culture base, is prepared according to the following steps；A, radix pseudostellariae seed is first sterilized with alcohol, then is sterilized with mercuric chloride, obtain A product；B, A product are selected into superclean bench complete embryo, is inoculated in radix pseudostellariae Seed embryo culture base, obtains B product；C, B product are cultivated to get radix pseudostellariae seedling.The present invention has the germination rate that can improve the emergence of radix pseudostellariae embryo, and germination rate is up to 99%, shortens seedling-growing time, and emergence is neat, and this method can quickly provide a large amount of radix pseudostellariae seedlings, promotes radix pseudostellariae artificial growth, meets the market demand.
Description
Technical field
The present invention relates to a kind of culture medium, especially a kind of radix pseudostellariae Seed embryo culture base and its cultural method.
Background technique
Radix pseudostellariae is Caryophyllaceae radix pseudostellariae category (Pseudostellaria) plant.It is traditional Chinese medicine common drug, medicine
Power is gentle, has and replenishes qi to invigorate the spleen, is promoting production of body fluid and nourishing the lung and other effects.Radix pseudostellariae is in tcm clinical practice formula, Chinese patent drug, health medicine production
Medicinal material used is all from cultivation resource.But for a long time, radix pseudostellariae carries out vegetative propagation with root tuber, causes cylinder accumulation big
Amount virus, seriously affects its yield.People are in cultivation, and also trial is bred with seed, but radix pseudostellariae seed has not
Moultinism.Under field conditions (factors), radix pseudostellariae seed sprouting needs about one year.Releasing its dormant trait using (5 DEG C) storages of low temperature also needs
30-60 days are taken, and germination rate is not high.Scholar's Ye Zuyun discovery radix pseudostellariae embryo tissue culture can grow complete seedling, but transport
It is cultivated with the culture medium of Ye Zuyun, radix pseudostellariae kind germination rate is not high.
Summary of the invention
The purpose of the present invention is to provide a kind of radix pseudostellariae Seed embryo culture base and its cultural method.The present invention has and can improve
The germination rate of radix pseudostellariae embryo emergence, germination rate are up to 99%, shorten seedling-growing time, emergence is neat, and this method can be mentioned quickly
For a large amount of radix pseudostellariae seedlings, promotes radix pseudostellariae artificial growth, meet the market demand.
The invention is realized in this way.A kind of radix pseudostellariae Seed embryo culture base, the radix pseudostellariae Seed embryo culture base are as follows: every liter of MS
Contain sucrose 28-32g, agar powder 7.2-7.8g, 6- benzyl aminoadenine 0.40-0.60mg, methyl α-naphthyl acetate 0.10- in culture medium
0.30mg and heteroauxin 0.15-0.25mg.
In radix pseudostellariae Seed embryo culture base above-mentioned, the radix pseudostellariae Seed embryo culture base are as follows: contain sugarcane in every liter of MS culture medium
Sugared 29-31g, agar powder 7.4-7.6g, 6- benzyl aminoadenine 0.45-0.55mg, methyl α-naphthyl acetate 0.15-0.25mg and heteroauxin
0.18-0.23mg。
In radix pseudostellariae Seed embryo culture base above-mentioned, the radix pseudostellariae Seed embryo culture base are as follows: contain sugarcane in every liter of MS culture medium
Sugared 30g, agar powder 7.5g, 6- benzyl aminoadenine 0.50mg, methyl α-naphthyl acetate 0.20mg and heteroauxin 0.20mg.
In radix pseudostellariae Seed embryo culture base above-mentioned, the pH value of the radix pseudostellariae Seed embryo culture base is 5.6-6.0.
In the cultural method of radix pseudostellariae Seed embryo culture base above-mentioned, it is prepared according to the following steps；
A, radix pseudostellariae seed is first sterilized with alcohol, then is sterilized with mercuric chloride, obtain A product；
B, A product are selected into superclean bench complete embryo, is inoculated in radix pseudostellariae Seed embryo culture base, obtains B product；
C, B product are cultivated to get radix pseudostellariae seedling.
It is that radix pseudostellariae seed is first used into 70- in the step a in the cultural method of radix pseudostellariae Seed embryo culture base above-mentioned
75% alcohol sterilizing 25-35s, then sterilizing 15-25min is carried out with the mercuric chloride of 0.1-0.2%.
In the cultural method of radix pseudostellariae Seed embryo culture base above-mentioned, in the step c, the temperature of culture is 14-17 DEG C.
In the cultural method of radix pseudostellariae Seed embryo culture base above-mentioned, in the step c, the intensity of illumination of culture is 1800-
2200Lx, illumination 11-13h/d.
Compared with the prior art, 6- benzyl aminoadenine main function is the shape for promoting bud in radix pseudostellariae Seed embryo culture base
At evoked callus occurs；Methyl α-naphthyl acetate can promote cell division and expand, induced synthesis adventitious root；Heteroauxin induction is thin
The division of born of the same parents, the differentiation for promoting root promote growth and take root so adding heteroauxin in culture medium；MS solid medium is available
Carry out evoked callus, or is used for embryo, stem section, stem apex and Anther Culture；Sucrose except as in culture medium carbon source and the energy in addition to,
It also plays an important role to the osmotic pressure for maintaining culture medium；Agar steady chemical structure is not easy to decompose utilization by mycelia, in culture medium
In only play coagulator.
Experiments have shown that:
1, applicant is inoculated with 100 radix pseudostellariae kinds using 1-5 of the embodiment of the present invention respectively and cultivates, and observes embryo germination
Situation grows to 3cm or so in rudiment and counts its germination rate, as a result as shown in the table；
1 radix pseudostellariae Seed embryo culture base germination rate of table
| Group | Cultivate quantity | Sprouting quantity | Germination rate |
| Embodiment 1 | 100 | 99 | 99% |
| Embodiment 2 | 100 | 98 | 98% |
| Embodiment 3 | 100 | 99 | 99% |
| Embodiment 4 | 100 | 97 | 97% |
| Embodiment 5 | 100 | 98 | 98% |
And in above embodiments 1-5, after cultivating 3d, cotyledon starts to expand radix pseudostellariae kind, and plumule starts to sprout,
3cm can be grown to after 10d.
2, the culture medium that applicant changes the radix pseudostellariae Seed embryo culture base in 1-5 of the embodiment of the present invention into Ye Zuyun is (every
Rise MS culture medium in contain 6- benzyl aminoadenine 1.0mg, methyl α-naphthyl acetate 0.2mg/L) after, respectively be inoculated with 100 radix pseudostellariae kinds into
Row culture, observes embryo germination situation, grows to 3cm or so in rudiment and count its germination rate, as a result as shown in the table；
The culture medium germination rate of 2 Ye Zuyun of table
| Group | Cultivate quantity | Sprouting quantity | Germination rate |
| Embodiment 1 | 100 | 74 | 74% |
| Embodiment 2 | 100 | 75 | 75% |
| Embodiment 3 | 100 | 76 | 76% |
| Embodiment 4 | 100 | 73 | 73% |
| Embodiment 5 | 100 | 77 | 77% |
And in above embodiments 1-5, after cultivating 7d, cotyledon starts to expand radix pseudostellariae kind, and plumule starts to sprout,
3cm can be grown to after 18d.
By table 1 and table 2 as can be seen that the present invention has the germination rate that can improve the emergence of radix pseudostellariae embryo, germination rate is high
Up to 99%, shorten seedling-growing time, emergence is neat.
In conclusion the present invention has the germination rate that can improve the emergence of radix pseudostellariae embryo, germination rate is up to 99%, shortens out
The seedling time, emergence is neat, and this method can quickly provide a large amount of radix pseudostellariae seedlings, promotes radix pseudostellariae artificial growth, meets market
Demand.
Specific embodiment
Embodiment 1.A kind of radix pseudostellariae Seed embryo culture base, the radix pseudostellariae Seed embryo culture base are as follows: contain in every liter of MS culture medium
There are sucrose 28g, agar powder 7.2g, 6- benzyl aminoadenine 0.40mg, methyl α-naphthyl acetate 0.10mg and heteroauxin 0.15mg.
The pH value of the radix pseudostellariae Seed embryo culture base is 5.6.
The cultural method of radix pseudostellariae Seed embryo culture base, is prepared according to the following steps；
A, radix pseudostellariae seed is first used to 70% alcohol sterilizing 25s, then carries out sterilizing 15min with 0.1% mercuric chloride, obtains A
Product；
B, A product are selected into complete embryo with pincet and scalpel in superclean bench, is inoculated in radix pseudostellariae embryo
In culture medium, B product are obtained；
C, B product to be cultivated, the temperature of culture is 14 DEG C, and the intensity of illumination of culture is 1800Lx, illumination 11h/d,
Up to radix pseudostellariae seedling.
Embodiment 2.A kind of radix pseudostellariae Seed embryo culture base, the radix pseudostellariae Seed embryo culture base are as follows: contain in every liter of MS culture medium
There are sucrose 29g, agar powder 7.4g, 6- benzyl aminoadenine 0.45mg, methyl α-naphthyl acetate 0.15mg and heteroauxin 0.18mg.
The pH value of the radix pseudostellariae Seed embryo culture base is 5.7.
The cultural method of radix pseudostellariae Seed embryo culture base, is prepared according to the following steps；
A, radix pseudostellariae seed is first used to 72% alcohol sterilizing 28s, then carries out sterilizing 17min with 0.12% mercuric chloride, obtains A
Product；
B, A product are selected into complete embryo with pincet and scalpel in superclean bench, is inoculated in radix pseudostellariae embryo
In culture medium, B product are obtained；
C, B product to be cultivated, the temperature of culture is 15 DEG C, and the intensity of illumination of culture is 1900Lx, illumination 12h/d,
Up to radix pseudostellariae seedling.
Embodiment 3.A kind of radix pseudostellariae Seed embryo culture base, the radix pseudostellariae Seed embryo culture base are as follows: contain in every liter of MS culture medium
There are sucrose 30g, agar powder 7.4g, 6- benzyl aminoadenine 0.50mg, methyl α-naphthyl acetate 0.20mg and heteroauxin 0.20mg.
The pH value of the radix pseudostellariae Seed embryo culture base is 5.8.
The cultural method of radix pseudostellariae Seed embryo culture base, is prepared according to the following steps；
A, radix pseudostellariae seed is first used to 73% alcohol sterilizing 30s, then carries out sterilizing 20min with 0.15% mercuric chloride, obtains A
Product；
B, A product are selected into complete embryo with pincet and scalpel in superclean bench, is inoculated in radix pseudostellariae embryo
In culture medium, B product are obtained；
C, B product to be cultivated, the temperature of culture is 15 DEG C, and the intensity of illumination of culture is 2000Lx, illumination 12h/d,
Up to radix pseudostellariae seedling.
Embodiment 4.A kind of radix pseudostellariae Seed embryo culture base, the radix pseudostellariae Seed embryo culture base are as follows: contain in every liter of MS culture medium
There are sucrose 31g, agar powder 7.6g, 6- benzyl aminoadenine 0.55mg, methyl α-naphthyl acetate 0.25mg and heteroauxin 0.23mg.
The pH value of the radix pseudostellariae Seed embryo culture base is 5.9.
The cultural method of radix pseudostellariae Seed embryo culture base, is prepared according to the following steps；
A, radix pseudostellariae seed is first used to 74% alcohol sterilizing 33s, then carries out sterilizing 22min with 0.18% mercuric chloride, obtains A
Product；
B, A product are selected into complete embryo with pincet and scalpel in superclean bench, is inoculated in radix pseudostellariae embryo
In culture medium, B product are obtained；
C, B product to be cultivated, the temperature of culture is 16 DEG C, and the intensity of illumination of culture is 2100Lx, illumination 12h/d,
Up to radix pseudostellariae seedling.
Embodiment 5.A kind of radix pseudostellariae Seed embryo culture base, the radix pseudostellariae Seed embryo culture base are as follows: contain in every liter of MS culture medium
There are sucrose 32g, agar powder 7.8g, 6- benzyl aminoadenine 0.60mg, methyl α-naphthyl acetate 0.30mg and heteroauxin 0.25mg.
The pH value of the radix pseudostellariae Seed embryo culture base is 6.0.
The cultural method of radix pseudostellariae Seed embryo culture base, is prepared according to the following steps；
A, radix pseudostellariae seed is first used to 75% alcohol sterilizing 35s, then carries out sterilizing 25min with 0.2% mercuric chloride, obtains A
Product；
B, A product are selected into complete embryo with pincet and scalpel in superclean bench, is inoculated in radix pseudostellariae embryo
In culture medium, B product are obtained；
C, B product to be cultivated, the temperature of culture is 17 DEG C, and the intensity of illumination of culture is 2200Lx, illumination 13h/d,
Up to radix pseudostellariae seedling.
Claims (8)
1. a kind of radix pseudostellariae Seed embryo culture base, it is characterised in that: the radix pseudostellariae Seed embryo culture base are as follows: contain in every liter of MS culture medium
There are sucrose 28-32g, agar powder 7.2-7.8g, 6- benzyl aminoadenine 0.40-0.60mg, methyl α-naphthyl acetate 0.10-0.30mg and indoles
Acetic acid 0.15-0.25mg.
2. radix pseudostellariae Seed embryo culture base according to claim 1, it is characterised in that: the radix pseudostellariae Seed embryo culture base are as follows:
Contain sucrose 29-31g, agar powder 7.4-7.6g, 6- benzyl aminoadenine 0.45-0.55mg, methyl α-naphthyl acetate in every liter of MS culture medium
0.15-0.25mg and heteroauxin 0.18-0.23mg.
3. radix pseudostellariae Seed embryo culture base according to claim 1, it is characterised in that: the radix pseudostellariae Seed embryo culture base are as follows:
Contain sucrose 30g, agar powder 7.5g, 6- benzyl aminoadenine 0.50mg, methyl α-naphthyl acetate 0.20mg and indoles in every liter of MS culture medium
Acetic acid 0.20mg.
4. radix pseudostellariae Seed embryo culture base according to claim 1, it is characterised in that: the pH of the radix pseudostellariae Seed embryo culture base
Value is 5.6-6.0.
5. a kind of cultural method of the radix pseudostellariae Seed embryo culture base as described in claim 1-4 any one, it is characterised in that: press
Following step preparation；
A, radix pseudostellariae seed is first sterilized with alcohol, then is sterilized with mercuric chloride, obtain A product；
B, A product are selected into superclean bench complete embryo, is inoculated in radix pseudostellariae Seed embryo culture base, obtains B product；
C, B product are cultivated to get radix pseudostellariae seedling.
6. the cultural method of radix pseudostellariae Seed embryo culture base according to claim 5, it is characterised in that: in the step a, be
Radix pseudostellariae seed is first used to the alcohol sterilizing 25-35s of 70-75%, then carries out sterilizing 15-25min with the mercuric chloride of 0.1-0.2%.
7. the cultural method of radix pseudostellariae Seed embryo culture base according to claim 5, it is characterised in that: in the step c, training
Feeding temperature is 14-17 DEG C.
8. the cultural method of radix pseudostellariae Seed embryo culture base according to claim 5, it is characterised in that: in the step c, training
Feeding intensity of illumination is 1800-2200Lx, illumination 11-13h/d.
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