CN107821160A - A kind of method for culturing seedlings of the bletilla striata - Google Patents
A kind of method for culturing seedlings of the bletilla striata Download PDFInfo
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- CN107821160A CN107821160A CN201711013640.6A CN201711013640A CN107821160A CN 107821160 A CN107821160 A CN 107821160A CN 201711013640 A CN201711013640 A CN 201711013640A CN 107821160 A CN107821160 A CN 107821160A
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- bletilla striata
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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
-
- 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
- A01H4/005—Methods for micropropagation; Vegetative plant propagation using cell or tissue culture techniques
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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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- Developmental Biology & Embryology (AREA)
- Engineering & Computer Science (AREA)
- Biotechnology (AREA)
- Cell Biology (AREA)
- Botany (AREA)
- Environmental Sciences (AREA)
- Breeding Of Plants And Reproduction By Means Of Culturing (AREA)
Abstract
The invention belongs to bletilla striata planting technology field, especially a kind of method for culturing seedlings of the bletilla striata, including explant processing, culture, second incubation and hardening totally four steps, bletilla striata lateral bud and stem apex are used as explant, it is mixed after treatment, in conjunction with the regulation and control to culture environment, and then strengthen the proliferation function of the bletilla striata, lift germination percentage, again after second incubation, so that plant is bud green, root system is sturdy and length is uniform, survival rate is high, on-bladed yellow, again after hardening treatment, so that the survival rate in bletilla striata transplantation of seedlings crop field is to more than 98%, and seedling robust growth, the speed of growth is fast.
Description
Technical field
The invention belongs to bletilla striata planting technology field, especially a kind of method for culturing seedlings of the bletilla striata.
Background technology
The bletilla striata is orchid family bletilla striata category herbaceos perennial, also makees bletilla, Lian Jicao, Gan Gen, be distributed in China to Korea,
The ground such as East China, south China and Henan, Shaanxi, Sichuan, Yunnan are originated in Japan, China, and the stem tuber of the bletilla striata is China's traditional Chinese medicine, early in
《Sheng Nong's herbal classic》In just it is on the books,《Chinese Pharmacopoeia》It is middle to record with tonifying lung, detumescence, myogenic, hemostasis, sore and other effects,
Hinder the diseases such as hemoptysis, metal-inflicted wound bleeding, canker sores, soup fire burn, rhagadia manus et pedis for treating lung；In addition, bletilla striata pattern is gorgeous,
It is a kind of good landscape plant, therefore market is very vigorous to the demand of the bletilla striata.
Due to the expansion of the market demand, the wild bletilla striata of China's most area is excessively excavated in recent years, savings amount
Reduction drastically, and the bletilla striata is not easy to germinate, and is mainly bred in a manner of the plant division being divided into two, not only low reproduction rate, breeding
Speed it is slow, and it is big to consume kind of amount, also fights for raw material with medication；Using tissue culture technique, can with a large amount of seedlings of fast culture,
To meet the needs of artificial cultivation, a kind of bletilla striata tissue method for culturing seedlings as disclosed in patent CN201510733897.3, it is operated
Simply, production cost is low, and the bletilla striata survival rate of cultivation is high, but its yield is still not ideal enough, and the cultivation effect of plant has much room for improvement,
It is possible to there is the phenomenon of yellowing leaf in incubation.
The content of the invention
In order to solve the above technical problems, the present invention provides a kind of method for culturing seedlings of the bletilla striata.
It is achieved particular by following technical scheme：
A kind of method for culturing seedlings of the bletilla striata, comprises the following steps：
(1) explant is handled：Fresh bletilla striata lateral bud and stem apex are selected, 1~2cm length is cut into, 2 is cleaned with distilled water
~3 times, then use concentration to soak 10~15min for 0.01~0.05% liquor potassic permanganate, then with sterile water wash 2~3
It is secondary, the moisture on surface is blotted with aseptic filter paper, and 0.2~0.5cm designated port is marked with cutter on the surface of lateral bud and stem apex,
The designated port number of each lateral bud and stem apex is 2~3；
(2) cultivate：The explant handled well is inoculated in inducing culture and cultivated one week, cultivation temperature is 26~28
DEG C, then move into that intensity of illumination is 2100~2300lx, light application time is 12~15h/d, humidity is 82~86%, temperature 23
18~22d is cultivated under the conditions of~25 DEG C；
(3) second incubation：The budlet that step (2) is turned out is inoculated into MS 30~35d of medium culture, in incubation
Temperature is 23~25 DEG C, and intensity of illumination is 1500~1800lx, and light application time is 12~15h/d, humidity is 85~95%；
(4) hardening：After second incubation terminates, by height for 4~5cm bletilla striata transplantation of seedlings to hardening in warmhouse booth 7~
8d, then be transplanted to and grow young leaves to bletilla striata seedling top containing culture in the native matrix in sepiolite, deer natural pond, bottom grows new root, stem
Portion, which significantly increases, may move into field planting.
The inducing culture be using MS culture mediums as minimal medium, and add 6-benzyl aminopurine, the basic element of cell division,
Methyl α-naphthyl acetate sodium salt, sucrose, agar powder.
2.0~the 2.5mg/L of 6-benzyl aminopurine, 1.5~2.0mg/L of the basic element of cell division, methyl α-naphthyl acetate sodium salt 0.1~
0.5mg/L, 20~25g/L of sucrose, 6~8g/L of agar powder.
The nutrient medium be using MS culture mediums as minimal medium, and add phoenix tree leaf powder, methyl α-naphthyl acetate, wood fragments bits,
Sucrose and agar powder.
40~50g/L of the phoenix tree leaf powder, 0.2~0.4mg/L of methyl α-naphthyl acetate, wood fragments consider 5~10g/L, 20~25g/ of sucrose to be worth doing
L, 3~5g/L of agar powder.
The weight proportion of the sepiolite and deer natural pond soil is 1：3.
In step (4) the transplanting incubation, 1 carbendazim, 800~900 times of liquid are sprayed week about, every two weeks
Spray the nutrient solution of a MS a great number of elements dilution 800~900.
Beneficial effect
Bletilla striata method for culturing seedlings of the present invention includes explant processing, culture, second incubation and hardening totally four steps, using white
Splendid achnatherum lateral bud and stem apex are explant, are mixed after treatment, in conjunction with the regulation and control to culture environment, and then strengthen the bletilla striata
Proliferation function, germination percentage is lifted, then after second incubation so that plant is bud green, root system is sturdy and length is uniform, and survival rate is high,
On-bladed yellow, then after hardening treatment so that the survival rate in bletilla striata transplantation of seedlings crop field to more than 98%, and seedling growth is strong
Strong, the speed of growth is fast.
Embodiment
Tuberculosis specific embodiment is limited technical scheme is further below, but claimed
Scope is not only limited to made description.
Embodiment 1
A kind of method for culturing seedlings of the bletilla striata, comprises the following steps：
(1) explant is handled：Fresh bletilla striata lateral bud and stem apex are selected, 1cm length is cut into, is cleaned 2 times with distilled water,
Use concentration to soak 10min for 0.01% liquor potassic permanganate again, then with sterile water wash 2 times, table is blotted with aseptic filter paper
The moisture in face, and 0.2cm designated port, the designated port number of each lateral bud and stem apex are marked on the surface of lateral bud and stem apex with cutter
For 2；
(2) cultivate：The explant handled well is inoculated in inducing culture and cultivated one week, cultivation temperature is 26 DEG C, so
Afterwards move into intensity of illumination be 2100lx, light application time 12h/d, humidity 82%, temperature cultivate 18d under the conditions of being 23 DEG C；
(3) second incubation：The budlet that step (2) is turned out is inoculated into MS medium culture 30d, temperature in incubation
For 23 DEG C, intensity of illumination 1500lx, light application time 12h/d, humidity 85%；
(4) hardening：After second incubation terminates, by height for 4cm bletilla striata transplantation of seedlings to hardening 7d in warmhouse booth, then move
Plant and grow young leaves to bletilla striata seedling top containing culture in the native matrix in sepiolite, deer natural pond, bottom grows new root, and stem substantially increases
It is big to may move into field planting.
The inducing culture be using MS culture mediums as minimal medium, and add 6-benzyl aminopurine, the basic element of cell division,
Methyl α-naphthyl acetate sodium salt, sucrose, agar powder.
The 6-benzyl aminopurine 2.0mg/L, basic element of cell division 1.5mg/L, methyl α-naphthyl acetate sodium salt 0.1mg/L, sucrose 20g/
L, agar powder 6g/L.
The nutrient medium be using MS culture mediums as minimal medium, and add phoenix tree leaf powder, methyl α-naphthyl acetate, wood fragments bits,
Sucrose and agar powder.
The phoenix tree leaf powder 40g/L, methyl α-naphthyl acetate 0.2mg/L, wood fragments bits 5g/L, sucrose 20g/L, agar powder 3g/L.
The weight proportion of the sepiolite and deer natural pond soil is 1：3.
In step (4) the transplanting incubation, 1 carbendazim, 800 times of liquid are sprayed week about, one was sprayed every two weeks
The nutrient solution of secondary MS a great number of elements dilution 800.
Control group 1 uses the method for culturing seedlings of the bletilla striata of embodiment 1, and phoenix tree leaf powder is not added during second incubation；
Control group 2 uses the method for culturing seedlings of the bletilla striata of embodiment 1, does not add wood fragments bits during second incubation；
Control group 3 uses the method for culturing seedlings of the bletilla striata of embodiment 1, and phoenix tree leaf powder and wood fragments are not added during second incubation
Bits.
Control group 4 organizes method for culturing seedlings using the patent CN201510733897.3 bletilla striata；
Experiment is divided into 5 groups, is inoculated with respectively according to above-mentioned method for culturing seedlings, every group of 10 plants of inoculation, after off-test, dialogue
The cultivation effect of splendid achnatherum, average plant height and growing state are measured, as a result such as following table：
From result in table, optimal using the scheme bletilla striata nursery effect of embodiment 1, cultivation effect is best, average plant height
3.5cm, and all long root of bletilla striata plant are reached, mean elements 10.3, plant is dark green；Therefore, by adding in the medium
Phoenix tree leaf powder and wood fragments bits are favorably improved the cultivation effect of the bletilla striata, promote the development of plant root, prevent plant leaf from occurring
Yellow.
Embodiment 2
A kind of method for culturing seedlings of the bletilla striata, comprises the following steps：
(1) explant is handled：Fresh bletilla striata lateral bud and stem apex are selected, 2cm length is cut into, is cleaned 3 times with distilled water,
Use concentration to soak 15min for 0.05% liquor potassic permanganate again, then with sterile water wash 3 times, table is blotted with aseptic filter paper
The moisture in face, and 0.5cm designated port, the designated port number of each lateral bud and stem apex are marked on the surface of lateral bud and stem apex with cutter
For 3；
(2) cultivate：The explant handled well is inoculated in inducing culture and cultivated one week, cultivation temperature is 28 DEG C, so
Afterwards move into intensity of illumination be 2300lx, light application time 15h/d, humidity 86%, temperature cultivate 22d under the conditions of being 25 DEG C；
(3) second incubation：The budlet that step (2) is turned out is inoculated into MS medium culture 35d, temperature in incubation
For 25 DEG C, intensity of illumination 1800lx, light application time 15h/d, humidity 95%；
(4) hardening：After second incubation terminates, by height for 5cm bletilla striata transplantation of seedlings to hardening 8d in warmhouse booth, then move
Plant and grow young leaves to bletilla striata seedling top containing culture in the native matrix in sepiolite, deer natural pond, bottom grows new root, and stem substantially increases
It is big to may move into field planting.
The inducing culture be using MS culture mediums as minimal medium, and add 6-benzyl aminopurine, the basic element of cell division,
Methyl α-naphthyl acetate sodium salt, sucrose, agar powder.
The 6-benzyl aminopurine 2.5mg/L, basic element of cell division 2.0mg/L, methyl α-naphthyl acetate sodium salt 0.5mg/L, sucrose 25g/
L, agar powder 8g/L.
The nutrient medium be using MS culture mediums as minimal medium, and add phoenix tree leaf powder, methyl α-naphthyl acetate, wood fragments bits,
Sucrose and agar powder.
The phoenix tree leaf powder 50g/L, methyl α-naphthyl acetate 0.4mg/L, wood fragments bits 10g/L, sucrose 25g/L, agar powder 5g/L.
The weight proportion of the sepiolite and deer natural pond soil is 1：3.
In step (4) the transplanting incubation, 1 carbendazim, 900 times of liquid are sprayed week about, one was sprayed every two weeks
The nutrient solution of secondary MS a great number of elements dilution 900.
Embodiment 3
A kind of method for culturing seedlings of the bletilla striata, comprises the following steps：
(1) explant is handled：Fresh bletilla striata lateral bud and stem apex are selected, 1cm length is cut into, is cleaned 3 times with distilled water,
Use concentration to soak 10min for 0.03% liquor potassic permanganate again, then with sterile water wash 3 times, table is blotted with aseptic filter paper
The moisture in face, and 0.4cm designated port, the designated port number of each lateral bud and stem apex are marked on the surface of lateral bud and stem apex with cutter
For 2；
(2) cultivate：The explant handled well is inoculated in inducing culture and cultivated one week, cultivation temperature is 27 DEG C, so
Afterwards move into intensity of illumination be 2200lx, light application time 14h/d, humidity 85%, temperature cultivate 20d under the conditions of being 24 DEG C；
(3) second incubation：The budlet that step (2) is turned out is inoculated into MS medium culture 33d, temperature in incubation
For 25 DEG C, intensity of illumination 1700lx, light application time 14h/d, humidity 90%；
(4) hardening：After second incubation terminates, by height for 4.5cm bletilla striata transplantation of seedlings to hardening 7d in warmhouse booth, then
It is transplanted to and grows young leaves to bletilla striata seedling top containing culture in the native matrix in sepiolite, deer natural pond, bottom grows new root, and stem is obvious
Increase may move into field planting.
The inducing culture be using MS culture mediums as minimal medium, and add 6-benzyl aminopurine, the basic element of cell division,
Methyl α-naphthyl acetate sodium salt, sucrose, agar powder.
The 6-benzyl aminopurine 2.3mg/L, basic element of cell division 1.7mg/L, methyl α-naphthyl acetate sodium salt 0.4mg/L, sucrose 23g/
L, agar powder 7g/L.
The nutrient medium be using MS culture mediums as minimal medium, and add phoenix tree leaf powder, methyl α-naphthyl acetate, wood fragments bits,
Sucrose and agar powder.
The phoenix tree leaf powder 46g/L, methyl α-naphthyl acetate 0.3mg/L, wood fragments bits 7g/L, sucrose 23g/L, agar powder 4g/L.
The weight proportion of the sepiolite and deer natural pond soil is 1：3.
In step (4) the transplanting incubation, 1 carbendazim, 850 times of liquid are sprayed week about, one was sprayed every two weeks
The nutrient solution of secondary MS a great number of elements dilution 850.
It is important to point out that, above example and test example are only limitted to do further technical scheme herein
Elaboration and understanding, it is impossible to be interpreted as further to technical scheme and limited, what those skilled in the art made
Non-protruding essential characteristics and the innovation and creation of marked improvement, still fall within the protection category of the present invention.
Claims (7)
1. a kind of method for culturing seedlings of the bletilla striata, it is characterised in that comprise the following steps：
(1) explant is handled：Fresh bletilla striata lateral bud and stem apex are selected, 1~2cm length is cut into, 2~3 is cleaned with distilled water
It is secondary, then use concentration to soak 10~15min for 0.01~0.05% liquor potassic permanganate, then with sterile water wash 2~3 times,
The moisture on surface is blotted with aseptic filter paper, and marks 0.2~0.5cm designated port with cutter on the surface of lateral bud and stem apex, often
The designated port number of individual lateral bud and stem apex is 2~3；
(2) cultivate：The explant handled well is inoculated in inducing culture and cultivated one week, cultivation temperature is 26~28 DEG C, so
Immigration intensity of illumination is 2100~2300lx afterwards, light application time is 12~15h/d, humidity is 82~86%, temperature is 23~25
18~22d is cultivated under the conditions of DEG C；
(3) second incubation：The budlet that step (2) is turned out is inoculated into MS 30~35d of medium culture, temperature in incubation
For 23~25 DEG C, intensity of illumination is 1500~1800lx, and light application time is 12~15h/d, humidity is 85~95%；
(4) hardening：After second incubation terminates, by height for 4~5cm bletilla striata transplantation of seedlings to 7~8d of hardening in warmhouse booth, then
It is transplanted to and grows young leaves to bletilla striata seedling top containing culture in the native matrix in sepiolite, deer natural pond, bottom grows new root, and stem is obvious
Increase may move into field planting.
2. the method for culturing seedlings of the bletilla striata as claimed in claim 1, it is characterised in that the inducing culture is using MS culture mediums as base
Basal culture medium, and add 6-benzyl aminopurine, the basic element of cell division, methyl α-naphthyl acetate sodium salt, sucrose, agar powder.
3. the method for culturing seedlings of the bletilla striata as claimed in claim 2, it is characterised in that the 2.0~2.5mg/L of 6-benzyl aminopurine,
1.5~2.0mg/L of the basic element of cell division, 0.1~0.5mg/L of methyl α-naphthyl acetate sodium salt, 20~25g/L of sucrose, 6~8g/L of agar powder.
4. the method for culturing seedlings of the bletilla striata as claimed in claim 1, it is characterised in that the nutrient medium is using MS culture mediums as base
Basal culture medium, and add phoenix tree leaf powder, methyl α-naphthyl acetate, wood fragments bits, sucrose and agar powder.
5. the method for culturing seedlings of the bletilla striata as claimed in claim 4, it is characterised in that 40~50g/L of the phoenix tree leaf powder, methyl α-naphthyl acetate
0.2~0.4mg/L, wood fragments consider 5~10g/L, 20~25g/L of sucrose, 3~5g/L of agar powder to be worth doing.
6. the method for culturing seedlings of the bletilla striata as claimed in claim 1, it is characterised in that the sepiolite and deer natural pond soil weight proportion be
1：3.
7. the method for culturing seedlings of the bletilla striata as claimed in claim 1, it is characterised in that in step (4) the transplanting incubation, every
1 carbendazim, 800~900 times of liquid are sprayed within one week, the nutrient solution of a MS a great number of elements dilution 800~900 was sprayed every two weeks.
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Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN108308029A (en) * | 2018-03-28 | 2018-07-24 | 普安县龙吟兴民种养殖开发有限公司 | A kind of bletilla striata tissue culture special culture media |
| CN108739387A (en) * | 2018-06-07 | 2018-11-06 | 金寨县金绿源特色中药材种植农场 | A kind of method for culturing seedlings of wild bletilla striata |
| CN109006292A (en) * | 2018-08-03 | 2018-12-18 | 遵义市龙驰生物科技有限公司 | A kind of quick seeding growing seedlings method of bletilla striata seeds simple plastic greenhouse |
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| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105104148A (en) * | 2015-09-16 | 2015-12-02 | 枞阳县白云生态园林有限责任公司 | Method for growing rhizoma atractylodis lanceae seedlings |
| CN106069100A (en) * | 2016-06-29 | 2016-11-09 | 安徽省固镇县谷阳有机蔬菜专业合作社 | A kind of implantation methods of Rhizoma Zingiberis Recens bud |
| CN106613936A (en) * | 2015-11-02 | 2017-05-10 | 丹阳华都园艺有限公司 | Method for raising Bletilla striata seedlings by tissue culture |
-
2017
- 2017-10-25 CN CN201711013640.6A patent/CN107821160A/en active Pending
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105104148A (en) * | 2015-09-16 | 2015-12-02 | 枞阳县白云生态园林有限责任公司 | Method for growing rhizoma atractylodis lanceae seedlings |
| CN106613936A (en) * | 2015-11-02 | 2017-05-10 | 丹阳华都园艺有限公司 | Method for raising Bletilla striata seedlings by tissue culture |
| CN106069100A (en) * | 2016-06-29 | 2016-11-09 | 安徽省固镇县谷阳有机蔬菜专业合作社 | A kind of implantation methods of Rhizoma Zingiberis Recens bud |
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN108308029A (en) * | 2018-03-28 | 2018-07-24 | 普安县龙吟兴民种养殖开发有限公司 | A kind of bletilla striata tissue culture special culture media |
| CN108739387A (en) * | 2018-06-07 | 2018-11-06 | 金寨县金绿源特色中药材种植农场 | A kind of method for culturing seedlings of wild bletilla striata |
| CN109006292A (en) * | 2018-08-03 | 2018-12-18 | 遵义市龙驰生物科技有限公司 | A kind of quick seeding growing seedlings method of bletilla striata seeds simple plastic greenhouse |
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Application publication date: 20180323 |