CN106613936A - Method for raising Bletilla striata seedlings by tissue culture - Google Patents
Method for raising Bletilla striata seedlings by tissue culture Download PDFInfo
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- CN106613936A CN106613936A CN201510733897.3A CN201510733897A CN106613936A CN 106613936 A CN106613936 A CN 106613936A CN 201510733897 A CN201510733897 A CN 201510733897A CN 106613936 A CN106613936 A CN 106613936A
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- bletilla striata
- tissue culture
- lateral bud
- bletilla
- culture
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Abstract
The invention relates to the technical field of seedling raising by tissue culture, in particular to a method for raising Bletilla striata seedlings by tissue culture; the method comprises the steps of preparing explants, culturing the explants in a medium, and transplanting. The method for culturing Bletilla striata by tissue culture is simple to perform and low in production cost, and survival rate of cultured Bletilla striata is high.
Description
Technical field
The present invention relates to the technical field of tissue culture, especially a kind of method of bletilla striata tissue culture.
Technical background
The bletilla striata, another name bletilla, pleionebulbocodioides rolfe, Pseudobulbus Bletillae (Rhizoma Bletillae), profit know son and company and grass etc., are that the orchid family bletilla striata belongs to for many years
Raw herbaceous plant.The effects such as its stem tuber has tonifying lung, detumescence, myogenic, hemostasis, sore, for treating lung
Hinder the diseases such as hemoptysis, canker sores, soup fire burn, brothers' cracking.In recent years, the bletilla striata is served not only as in tradition
Herbal medicine and ornamental plant extensive utilization, and be also increasingly subject to pay attention in the utilization field of cosmetics.At present,
The bletilla striata mainly to gather based on wild natural resources, due to the continuous expansion of operation strategies, its wild resource quilt
The excavation of transition, the reduction drastically of savings amount.The bletilla striata is difficult germination, mainly the plant division mode to be divided into two
Bred, not only low reproduction rate, the speed of breeding are slow, and consumption kind amount is big, also fights for raw material with medication.
Using tissue culture technique, can be with a large amount of seedlings of fast culture, to meet tame needs.
The content of the invention
The technical problem to be solved in the invention is a kind of bletilla striata tissue culture proposed for above-mentioned technical problem
Method.
To solve above-mentioned technical problem, the invention provides a kind of method of bletilla striata tissue culture, including it is as follows
Step：S1：The preparation of explant：Fresh bletilla striata lateral bud is chosen, first lateral bud is cleaned with distilled water,
Soak 10min～20min with liquid detergent solution again；Then bletilla striata lateral bud is cleaned with distilled water, is placed in 80%
Alcohol-pickled 1～2min, finally 10～15min of sterilizing is carried out with 0.2% raw mercury solution, after sterilizing
Bletilla striata lateral bud again with sterile water wash 3～5 times, cut away the bottom shape of bletilla striata lateral bud with the cutter sterilized
Into fresh wound；S2：Above-mentioned explant is put into MS culture mediums and is cultivated；S3：In the medium
Addition 0.1～0.4mg/L of 0.6～2.5mg/L of 6-benzyl aminopurine and methyl α-naphthyl acetate；S4：Work as bletilla tissue culture seedlings
When growing more than 3 roots on root media, by bottle seedling 3～5d of hardening disposed within, then bottle cap is opened,
1～2d of hardening, then removes culture medium by seedling, heels in 3 months in sand bed, is finally transplanted.
Further, in above-mentioned technical proposal, agar 5g/L, white sugar are additionally provided with the S2 in culture medium
20g/L。
Further, in above-mentioned technical proposal, the condition of culture in the S2 be intensity of illumination 1600～
2000lx, light application time is 15h/d, and cultivation temperature is 24 degree～28 degree.
Further, in above-mentioned technical proposal, the 6-benzyl aminopurine be 1.5mg/L, the methyl α-naphthyl acetate
For 0.1mg/L.
After the said method tissue culture bletilla striata, simple to operate, low production cost, the survival rate of the bletilla striata of cultivation
It is high.
Specific embodiment
Above-mentioned tissue culture method is all the optimum proportioning that inventor learns through many experiments, especially 6- benzyls amino
Purine is 1.5mg/L and methyl α-naphthyl acetate is 0.1mg/L.When the amount of 6-benzyl aminopurine addition is excessive, can lead
Inducing clumping bud rate is caused to decline, it is linear that bud mutation carefully dies down, and burnt tail is there is also when serious and is put into phenomenon.
When the amount addition of methyl α-naphthyl acetate is excessive, growth coefficient can be reduced, and the callus in bastem portion can increase. seedling
It is thin and delicate, leaf color jaundice.
Embodiment one：
S1：The preparation of explant：Fresh bletilla striata lateral bud is chosen, first lateral bud is cleaned with distilled water, then
18min is soaked with liquid detergent solution；Then bletilla striata lateral bud is cleaned with distilled water, be placed in 80% it is alcohol-pickled
1min, finally carries out sterilizing 12min with 0.2% raw mercury solution, and the bletilla striata lateral bud after sterilizing is again with aseptic
Water is cleaned 3 times, and fresh wound is formed on the bottom for cutting away bletilla striata lateral bud with the cutter sterilized；S2：Will be upper
The explant stated is put into MS culture mediums and is cultivated；S3：Add 6-benzyl aminopurine in the medium
0.8mg/L and methyl α-naphthyl acetate 0.2mg/L；S4：When bletilla tissue culture seedlings grow more than 3 roots on root media
When, by bottle seedling hardening 4d disposed within, then bottle cap is opened, then seedling is removed culture medium by hardening 1d,
Heel in 3 months in sand bed, finally transplanted.
100 seedling are test in experiment one, last test seedling percent is 86.8%.
Embodiment two：
S1：The preparation of explant：Fresh bletilla striata lateral bud is chosen, first lateral bud is cleaned with distilled water, then
15min is soaked with liquid detergent solution；Then bletilla striata lateral bud is cleaned with distilled water, be placed in 80% it is alcohol-pickled
2min, finally carries out sterilizing 13min with 0.2% raw mercury solution, and the bletilla striata lateral bud after sterilizing is again with aseptic
Water is cleaned 4 times, and fresh wound is formed on the bottom for cutting away bletilla striata lateral bud with the cutter sterilized；S2：Will be upper
The explant stated is put into MS culture mediums and is cultivated；S3：Add 6-benzyl aminopurine in the medium
1.5mg/L and methyl α-naphthyl acetate 0.1mg/L；S4：When bletilla tissue culture seedlings grow more than 3 roots on root media
When, by bottle seedling hardening 3d disposed within, then bottle cap is opened, then seedling is removed culture medium by hardening 1d,
Heel in 3 months in sand bed, finally transplanted.
100 seedling are test in experiment two, last test seedling percent is 96.5%.
Embodiment three：
S1：The preparation of explant：Fresh bletilla striata lateral bud is chosen, first lateral bud is cleaned with distilled water, then
20min is soaked with liquid detergent solution；Then bletilla striata lateral bud is cleaned with distilled water, be placed in 80% it is alcohol-pickled
2min, finally carries out sterilizing 15min with 0.2% raw mercury solution, and the bletilla striata lateral bud after sterilizing is again with aseptic
Water is cleaned 4 times, and fresh wound is formed on the bottom for cutting away bletilla striata lateral bud with the cutter sterilized；S2：Will be upper
The explant stated is put into MS culture mediums and is cultivated；S3：Add 6-benzyl aminopurine in the medium
2.3mg/L and methyl α-naphthyl acetate 0.3mg/L；S4：When bletilla tissue culture seedlings grow more than 3 roots on root media
When, by bottle seedling hardening 4d disposed within, then bottle cap is opened, then seedling is removed culture medium by hardening 1d,
Heel in 3 months in sand bed, finally transplanted.
100 seedling are test in embodiment three, last test seedling percent is 93.2%.
Can be known by above three embodiment, embodiment two is most preferred embodiment.The survival rate of bletilla striata seedling is most
Height, growing state is optimal.
Although the foregoing describing the specific embodiment of the present invention, those skilled in the art should manage
Solution, these are merely illustrative of, and can make various changes or modifications to present embodiment, without departing from this
The principle and essence of invention, protection scope of the present invention is only limited by the claims that follow.
Claims (3)
1. a kind of method of bletilla striata tissue culture, it is characterised in that comprise the steps：
S1：The preparation of explant：Fresh bletilla striata lateral bud is chosen, first lateral bud is cleaned with distilled water, then
10min～20min is soaked with liquid detergent solution；Then bletilla striata lateral bud is cleaned with distilled water, is placed in 80%
Alcohol-pickled 1～2min, finally carry out 10～15min of sterilizing with 0.2% raw mercury solution, through sterilizing
Bletilla striata lateral bud afterwards with sterile water wash 3～5 times, with the cutter sterilized bletilla striata lateral bud is cut away again
Fresh wound is formed on bottom；
S2：Above-mentioned explant is put into MS culture mediums and is cultivated；
S3：Add 0.1～0.4mg/L of 0.6～2.5mg/L of 6-benzyl aminopurine and methyl α-naphthyl acetate in the medium；
S4：When bletilla tissue culture seedlings grow more than 3 roots on root media, by bottle seedling hardening disposed within
3～5d, then bottle cap is opened, then seedling is removed culture medium by 1～2d of hardening, is heeled in sand
3 months in bed, finally transplanted.
2. the method for a kind of bletilla striata tissue culture according to claim 1, it is characterised in that：In the S2
Agar 5g/L, white sugar 20g/L are additionally provided with culture medium.
3. the method for a kind of bletilla striata tissue culture according to claim 1, it is characterised in that：In the S2
Condition of culture be 1600～2000lx of intensity of illumination, light application time is 15h/d, and cultivation temperature is 24
～28 degree of degree.
A kind of method of bletilla striata tissue culture according to claim 1, it is characterised in that：The 6- benzyls ammonia
Base purine is 1.5mg/L, and the methyl α-naphthyl acetate is 0.1mg/L.
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| Application Number | Priority Date | Filing Date | Title |
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| CN201510733897.3A CN106613936A (en) | 2015-11-02 | 2015-11-02 | Method for raising Bletilla striata seedlings by tissue culture |
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| CN201510733897.3A CN106613936A (en) | 2015-11-02 | 2015-11-02 | Method for raising Bletilla striata seedlings by tissue culture |
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Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN107182788A (en) * | 2017-07-11 | 2017-09-22 | 中国科学院武汉植物园 | The propagation method of bletilla seedling |
| CN107821160A (en) * | 2017-10-25 | 2018-03-23 | 石阡县龙腾农牧农民专业合作社 | A kind of method for culturing seedlings of the bletilla striata |
| CN108522184A (en) * | 2018-01-17 | 2018-09-14 | 刘雅文 | A kind of rapid propagation method for bletilla striata |
| CN109496709A (en) * | 2018-12-21 | 2019-03-22 | 广西壮族自治区农业科学院生物技术研究所 | A kind of method that bletilla tissue-cultured seedling temporary planting strong sprout promotees bud |
Citations (3)
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|---|---|---|---|---|
| CN102599063A (en) * | 2012-04-09 | 2012-07-25 | 向华 | Rapid propagation method of Bletilla striata |
| CN103314856A (en) * | 2013-07-08 | 2013-09-25 | 重庆市秀山红星中药材开发有限公司 | Bletilla lateral bud tissue culture propagation method |
| KR20150103879A (en) * | 2014-03-04 | 2015-09-14 | 고려대학교 산학협력단 | Producing method of orchid seedlings |
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2015
- 2015-11-02 CN CN201510733897.3A patent/CN106613936A/en active Pending
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102599063A (en) * | 2012-04-09 | 2012-07-25 | 向华 | Rapid propagation method of Bletilla striata |
| CN103314856A (en) * | 2013-07-08 | 2013-09-25 | 重庆市秀山红星中药材开发有限公司 | Bletilla lateral bud tissue culture propagation method |
| KR20150103879A (en) * | 2014-03-04 | 2015-09-14 | 고려대학교 산학협력단 | Producing method of orchid seedlings |
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Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN107182788A (en) * | 2017-07-11 | 2017-09-22 | 中国科学院武汉植物园 | The propagation method of bletilla seedling |
| CN107821160A (en) * | 2017-10-25 | 2018-03-23 | 石阡县龙腾农牧农民专业合作社 | A kind of method for culturing seedlings of the bletilla striata |
| CN108522184A (en) * | 2018-01-17 | 2018-09-14 | 刘雅文 | A kind of rapid propagation method for bletilla striata |
| CN109496709A (en) * | 2018-12-21 | 2019-03-22 | 广西壮族自治区农业科学院生物技术研究所 | A kind of method that bletilla tissue-cultured seedling temporary planting strong sprout promotees bud |
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