CN104686362A - Butterfly orchid tissue culturing method - Google Patents
Butterfly orchid tissue culturing method Download PDFInfo
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- CN104686362A CN104686362A CN201510128371.2A CN201510128371A CN104686362A CN 104686362 A CN104686362 A CN 104686362A CN 201510128371 A CN201510128371 A CN 201510128371A CN 104686362 A CN104686362 A CN 104686362A
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Abstract
The invention discloses a butterfly orchid tissue culturing method. The method comprises the steps of obtaining butterfly orchid stem tips, sterilizing the butterfly orchid stem tips, conducting preliminary culturing and subculturing to the butterfly orchid stem tips obtained after sterilization, and conducting rooting culturing. The method has the advantages that by means of the method, the rooting percentage of butterfly orchid tissue culture seedling plants reaches 26.7% at the maximum; tissue culture seedlings which are generated by the adoption of the stem tips and cultured in an isolated mode are capable of maintaining excellent characteristics of an original variety, so that foundations of elite propagation and industrial production are laid; according to the butterfly orchid tissue culturing method, the operation is simple, the cost is low, and the method is applicable to intermediate propagation of majority of ornamental butterfly orchids.
Description
Technical field
The present invention relates to plant tissue culture technique, especially relate to a kind of tissue culture method of Moth orchid.
Background technology
Moth orchid (Phalaenopsis aphrodite Rchb. F.) is the orchid family Phalaenopsis, and originating in hylaeion hypotropicum area, is growing nonparasitically upon another plant property orchid.The thick aerial root of Moth orchid white is exposed at around blade, except having the effect of nutrient in absorption air, also has growth and photosynthesis.In the time in the new year, Phalaenopsis plants extracts long long bennet out from axil, and output shape as butterfly dance in the air as flower, the dark favor by flower fans, have the title of " cattleya queen consort ".
At present, the rapid propagation in vitro research of Phalaenopsis plant focuses mostly on tree peony, about the domestic and international research of Moth orchid tissue cultures is less, existing about Moth orchid tissue culture technique, only discuss the foundation of sterile system, also not forming the method that a whole set of utilizes Moth orchid underground bud Fast-propagation, do not relate to taking root and transplanting of Moth orchid plantlet in vitro yet, foundation can not be provided for producing.
Summary of the invention
The technical solution adopted for the present invention to solve the technical problems is:
A tissue culture method for Moth orchid, comprises the acquisition of Moth orchid stem apex, Moth orchid stem apex sterilization and the Moth orchid stem apex after sterilization is carried out to the step of Initial culture, squamous subculture and culture of rootage, it is characterized in that:
(1) Moth orchid stem apex obtains: the budlet that clip butterfly in autumn Lanzhou and Xinjiang is sprouted, and is put on superclean bench after pure water flow wash 30min;
(2) Moth orchid stem apex sterilization: with alcohol-pickled 30 ~ 60 seconds of 70% ~ 75%, then with through water-reducible 1: 3 84 medicining liquid dipping 12 ~ 15 minutes, and blot surface moisture;
(3) Initial culture of Moth orchid stem apex:
1) squamous subculture is obtained plantlet in vitro 3 ~ 6 DEG C of refrigerations 8 ~ 10 days, be transferred on the root media containing IBA and cultivate 15 ~ 20 days;
2) plantlet in vitro is transferred on WPM+2% AC medium and continues cultivation 20 ~ 30 days, after taking root, carry out hardening;
3) transplantation of seedlings will be taken root in sphagna moss mixotrophism liquid matrix;
(4) Initial culture of Moth orchid:
Get budlet base portion, be cut into the segment that lcm is long, be inoculated on budlet inducing culture that pH value is 5.8-6.2;
The medium of described Initial culture is 1/2MS+0.1mg/L ~ 1mg/L GA3+1mg/L 6-BA+0.1mg/L ~ 0.5mg/L NAA+MN+2.0 mg/L;
The medium of described squamous subculture is 1/2MS+0.5mg/L ~ 1mg/L 6-BA, 1/2MS+0.1mg/L ~ 0.2mg/L TDZ, sucrose 30g/L, agar 6g/L,
(5) squamous subculture of Moth orchid:
First carry out dark treatment 5 weeks, then give illumination every day 24h, the cultivation temperature of Multiplying culture is 24-26 DEG C, and illumination is 30-40 μm of o1/m
2s;
(6) culture of rootage of Moth orchid is cultivated:
When root length is for 2-4cm, carries out rooting culture, blake bottle is moved to hardening 7-10 days in booth, wash away medium, root soaks 2-9% alcoholic solution and carries out disinfection, and be transplanted in the dish of cave, cover film, shades 75% simultaneously, remove film after 2 weeks, give full exposure, carry out rich water quality management.
Beneficial effect of the present invention:
(1) by Moth orchid plantlet in vitro plant of the present invention, rooting rate is up to 26.7%.
(2) plantlet in vitro adopting stem apex to carry out cultured in vitro generation can keep the merit of original kind, produces lay a good foundation for stock breeding and industrialization.
(3) iris tissue culture method of the present invention, simple to operate, with low cost, be suitable for the Fast-propagation that majority views and admires Moth orchid.
Embodiment
Embodiment 1:
A tissue culture method for Moth orchid, comprises the acquisition of Moth orchid stem apex, Moth orchid stem apex sterilization and the Moth orchid stem apex after sterilization is carried out to the step of Initial culture, squamous subculture and culture of rootage, it is characterized in that:
(1) Moth orchid stem apex obtains:
The budlet that clip butterfly in autumn Lanzhou and Xinjiang is sprouted, is put on superclean bench after pure water flow wash 30min;
(2) Moth orchid stem apex sterilization:
With alcohol-pickled 60 seconds of 70%, then with through water-reducible 1: 3 84 medicining liquid dipping 12 minutes, and blot surface moisture;
(3) Initial culture of Moth orchid stem apex:
1) squamous subculture is obtained plantlet in vitro 3 DEG C of refrigerations 8 days, be transferred on the root media containing IBA and cultivate 15 days;
2) plantlet in vitro is transferred on WPM+2% AC medium and continues cultivation 20 days, after taking root, carry out hardening;
3) transplantation of seedlings will be taken root in sphagna moss mixotrophism liquid matrix;
(4) Initial culture of Moth orchid:
Get budlet base portion, be cut into the segment that lcm is long, be inoculated on budlet inducing culture that pH value is 5.8-6.2;
The medium of described Initial culture is 1/2MS+0.1mg/L ~ 1mg/L GA3+1mg/L 6-BA+0.1mg/L ~ 0.5mg/L NAA+MN+2.0 mg/L;
The medium of described squamous subculture is 1/2MS+0.5mg/L ~ 1mg/L 6-BA, 1/2MS+0.1mg/L ~ 0.2mg/L TDZ, sucrose 30g/L, agar 6g/L,
(5) squamous subculture of Moth orchid:
First carry out dark treatment 5 weeks, then give illumination every day 24h, the cultivation temperature of Multiplying culture is 24-26 DEG C, and illumination is 30-40 μm of o1/m
2s;
(6) culture of rootage of Moth orchid is cultivated:
When root length is for 2-4cm, carries out rooting culture, blake bottle is moved to hardening 7-10 days in booth, wash away medium, root soaks 2-9% alcoholic solution and carries out disinfection, and be transplanted in the dish of cave, cover film, shades 75% simultaneously, remove film after 2 weeks, give full exposure, carry out rich water quality management.
Embodiment 2:
A tissue culture method for Moth orchid, comprises the acquisition of Moth orchid stem apex, Moth orchid stem apex obtains sterilization and the Moth orchid stem apex after sterilization is carried out to the step of Initial culture, squamous subculture and culture of rootage, it is characterized in that:
(1) Moth orchid stem apex obtains:
The budlet that clip butterfly in autumn Lanzhou and Xinjiang is sprouted, is put on superclean bench after pure water flow wash 30min;
(2) Moth orchid stem apex sterilization:
With alcohol-pickled 30 seconds of 75%, then with through water-reducible 1: 3 84 medicining liquid dipping 12 minutes, and blot surface moisture;
(3) Initial culture of Moth orchid stem apex:
1) squamous subculture is obtained plantlet in vitro 3 ~ 6 DEG C of refrigerations 8 ~ 10 days, be transferred on the root media containing IBA and cultivate 15 days;
2) plantlet in vitro is transferred on WPM+2% AC medium and continues cultivation 20 days, after taking root, carry out hardening;
3) transplantation of seedlings will be taken root in sphagna moss mixotrophism liquid matrix;
(4) Initial culture of Moth orchid:
Get budlet base portion, be cut into the segment that lcm is long, be inoculated on budlet inducing culture that pH value is 5.8-6.2;
The medium of described Initial culture is 1/2MS+0.1mg/L ~ 1mg/L GA3+1mg/L 6-BA+0.1mg/L ~ 0.5mg/L NAA+MN+2.0 mg/L;
The medium of described squamous subculture is 1/2MS+0.5mg/L ~ 1mg/L 6-BA, 1/2MS+0.1mg/L ~ 0.2mg/L TDZ, sucrose 30g/L, agar 6g/L,
(5) squamous subculture of Moth orchid:
First carry out dark treatment 5 weeks, then give illumination every day 24h, the cultivation temperature of Multiplying culture is 24-26 DEG C, and illumination is 30-40 μm of o1/m
2s;
(6) culture of rootage of Moth orchid is cultivated:
When root length is for 2-4cm, carries out rooting culture, blake bottle is moved to hardening 7-10 days in booth, wash away medium, root soaks 2-9% alcoholic solution and carries out disinfection, and be transplanted in the dish of cave, cover film, shades 75% simultaneously, remove film after 2 weeks, give full exposure, carry out rich water quality management.
Above embodiment is only be described the preferred embodiment of the present invention; not scope of the present invention is limited; under not departing from the present invention and designing the prerequisite of spirit; the various distortion that the common engineers and technicians in this area make technical scheme of the present invention and improvement, all should fall in protection domain that claims of the present invention determine.
The part that the present invention does not relate to prior art that maybe can adopt all same as the prior art is realized.
Claims (1)
1. a tissue culture method for Moth orchid, comprises the acquisition of Moth orchid stem apex, Moth orchid stem apex sterilization and the Moth orchid stem apex after sterilization is carried out to the step of Initial culture, squamous subculture and culture of rootage, it is characterized in that:
(1) Moth orchid stem apex obtains: the budlet that clip butterfly in autumn Lanzhou and Xinjiang is sprouted, and is put on superclean bench after pure water flow wash 30min;
(2) Moth orchid stem apex sterilization: with alcohol-pickled 30 ~ 60 seconds of 70% ~ 75%, then with through water-reducible 1: 3 84 medicining liquid dipping 12 ~ 15 minutes, and blot surface moisture;
(3) Initial culture of Moth orchid stem apex:
1) squamous subculture is obtained plantlet in vitro 3 ~ 6 DEG C of refrigerations 8 ~ 10 days, be transferred on the root media containing IBA and cultivate 15 ~ 20 days;
2) plantlet in vitro is transferred on WPM+2%AC medium and continues cultivation 20 ~ 30 days, after taking root, carry out hardening;
3) transplantation of seedlings will be taken root in sphagna moss mixotrophism liquid matrix;
(4) Initial culture of Moth orchid:
Get budlet base portion, be cut into the segment that lcm is long, be inoculated on budlet inducing culture that pH value is 5.8-6.2;
The medium of described Initial culture is 1/2MS+0.1mg/L ~ 1mg/L GA3+1mg/L 6-BA+0.1mg/L ~ 0.5mg/L NAA+MN+2.0 mg/L 6-BA+0.5 mg/L 2,4-D;
The medium of described squamous subculture is 1/2MS+0.5mg/L ~ 1mg/L6-BA, 1/2MS+0.1mg/L ~ 0.2mg/L TDZ, sucrose 30g/L, agar 6g/L,
(5) squamous subculture of Moth orchid:
First carry out dark treatment 5 weeks, then give illumination every day 24h, the cultivation temperature of Multiplying culture is 24-26 DEG C, and illumination is 30-40 μm of o1/m
2s;
(6) culture of rootage of Moth orchid is cultivated:
As the long 2-4cm of root, carry out rooting culture, blake bottle is moved to hardening 7-10 days in booth, wash away medium, root soaks 2-9% alcoholic solution and carries out disinfection, and be transplanted in the dish of cave, cover film, shades 75% simultaneously, remove film after 2 weeks, give full exposure, carry out rich water quality management.
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| CN201510128371.2A CN104686362A (en) | 2015-03-24 | 2015-03-24 | Butterfly orchid tissue culturing method |
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Cited By (10)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105941161A (en) * | 2016-06-25 | 2016-09-21 | 合肥隆扬农业科技有限公司 | Tissue culture method of phalaenopsis |
| CN106234179A (en) * | 2016-07-28 | 2016-12-21 | 张议亓 | A kind of method utilizing nutritional solution to cultivate iris |
| CN106472319A (en) * | 2016-10-20 | 2017-03-08 | 山东博华高效生态农业科技有限公司 | A kind of iris detoxification and fast breeding technique |
| CN107278901A (en) * | 2017-07-31 | 2017-10-24 | 王生旭 | A kind of sterilization method of iris bennet bud |
| CN107593457A (en) * | 2017-11-10 | 2018-01-19 | 佛山市恒爱网络科技有限公司 | A kind of iris subculture medium |
| CN107691221A (en) * | 2017-11-10 | 2018-02-16 | 佛山市恒爱网络科技有限公司 | A kind of iris tissue culture method |
| CN107810851A (en) * | 2017-11-10 | 2018-03-20 | 佛山市恒爱网络科技有限公司 | A kind of iris Initial culture base |
| CN108496753A (en) * | 2018-03-07 | 2018-09-07 | 山东省烟台市农业科学研究院 | A method of delaying iris cultivation matrix acidification corruption |
| CN109662010A (en) * | 2019-02-14 | 2019-04-23 | 芜湖东源新农村开发股份有限公司 | It is a kind of to prevent iris rotten culture substrate and preparation method thereof |
| CN109769646A (en) * | 2019-03-31 | 2019-05-21 | 芜湖东源新农村开发股份有限公司 | Extend the breeding method at iris florescence |
Citations (2)
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| FR2978160A1 (en) * | 2011-07-22 | 2013-01-25 | I R B Istituto Di Ricerche Biotecnologiche S R L | Cell culture, useful for preparing meristematic cells, which are useful e.g. as dietary supplement and as a drug, which has an antioxidant and anti-inflammatory activity, comprises phenylpropanoid and hydrosoluble polysaccharides |
| CN103004603A (en) * | 2012-12-27 | 2013-04-03 | 福建农林大学 | Plant regeneration method for butterfly orchid |
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2015
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Patent Citations (2)
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| FR2978160A1 (en) * | 2011-07-22 | 2013-01-25 | I R B Istituto Di Ricerche Biotecnologiche S R L | Cell culture, useful for preparing meristematic cells, which are useful e.g. as dietary supplement and as a drug, which has an antioxidant and anti-inflammatory activity, comprises phenylpropanoid and hydrosoluble polysaccharides |
| CN103004603A (en) * | 2012-12-27 | 2013-04-03 | 福建农林大学 | Plant regeneration method for butterfly orchid |
Non-Patent Citations (2)
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Cited By (12)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105941161A (en) * | 2016-06-25 | 2016-09-21 | 合肥隆扬农业科技有限公司 | Tissue culture method of phalaenopsis |
| CN106234179A (en) * | 2016-07-28 | 2016-12-21 | 张议亓 | A kind of method utilizing nutritional solution to cultivate iris |
| CN106472319A (en) * | 2016-10-20 | 2017-03-08 | 山东博华高效生态农业科技有限公司 | A kind of iris detoxification and fast breeding technique |
| CN106472319B (en) * | 2016-10-20 | 2018-10-26 | 山东博华高效生态农业科技有限公司 | A kind of iris detoxification and fast breeding technique |
| CN107278901A (en) * | 2017-07-31 | 2017-10-24 | 王生旭 | A kind of sterilization method of iris bennet bud |
| CN107593457A (en) * | 2017-11-10 | 2018-01-19 | 佛山市恒爱网络科技有限公司 | A kind of iris subculture medium |
| CN107691221A (en) * | 2017-11-10 | 2018-02-16 | 佛山市恒爱网络科技有限公司 | A kind of iris tissue culture method |
| CN107810851A (en) * | 2017-11-10 | 2018-03-20 | 佛山市恒爱网络科技有限公司 | A kind of iris Initial culture base |
| CN108496753A (en) * | 2018-03-07 | 2018-09-07 | 山东省烟台市农业科学研究院 | A method of delaying iris cultivation matrix acidification corruption |
| CN109662010A (en) * | 2019-02-14 | 2019-04-23 | 芜湖东源新农村开发股份有限公司 | It is a kind of to prevent iris rotten culture substrate and preparation method thereof |
| CN109769646A (en) * | 2019-03-31 | 2019-05-21 | 芜湖东源新农村开发股份有限公司 | Extend the breeding method at iris florescence |
| CN109769646B (en) * | 2019-03-31 | 2021-09-17 | 芜湖东源新农村开发股份有限公司 | Cultivation method for prolonging flowering phase of butterfly orchid |
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