CN106171994A - A kind of Sa Wanabai subculture method - Google Patents
A kind of Sa Wanabai subculture method Download PDFInfo
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- CN106171994A CN106171994A CN201610569338.8A CN201610569338A CN106171994A CN 106171994 A CN106171994 A CN 106171994A CN 201610569338 A CN201610569338 A CN 201610569338A CN 106171994 A CN106171994 A CN 106171994A
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- wanabai
- culture
- culture medium
- described step
- subculture method
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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
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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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- 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 discloses a kind of Sa Wanabai subculture method, comprise the following steps: the preparation of (1) culture medium: select basal medium, add sucrose and agar, add 6 benzyl aminoadenine and naphthalene acetic acids, regulate pH value, sterilizing, to obtain final product；(2) outer implant processes: the Sa Wanabai withered and yellow part of stem section in Primary culture cut off, and cleans；(3) inoculated and cultured: outer for step (2) the gained old wound of implant is cut off, in inserting step (1) gained culture medium, is placed in cultivation indoor cultivation.Relative to prior art, the inventive method not only has the advantage of tissue culture propagation in prior art, also by specific culture medium, hormone kind and concentration, combine with other processing method, it is possible to the proliferation times (1) significantly improving successive transfer culture reaches more than 2 times；(2) increasing the propagation algebraically of successive transfer culture, make originally can to breed generation can reach 10 generations more than, improves about more than 100%；(3) scale of Sa Wanabai, factorial praluction is made to be possibly realized.
Description
Technical field
The present invention relates to a kind of Sa Wanabai subculture method, belong to seedling propagation technical field.
Background technology
Sa Wanabai (Chamaecyparis pisifera cv.aurea) is also called golden coastline, gold leaf Japan spends
Cypress, gold leaf Sa Wana Cupressus funebris.Cupressaceae retinispora, belongs to the yellow kind in evergreen coloured silkization coniferous tree.Bark is that thin slice is
Bronzing.Scale leaf is close to, and sprig is sagging, elongated.March blooms, and female cone Dan Shengzhi pushes up, and male cone is oval, hermaphroditism, ball
Really November is ripe, and seed has the micro-flat oval of corner angle, and fruit scale peltate is wooden.Temperate zone and tropical trees, property happiness warm and moist gas
Wait, like light, cold-resistant drought-enduring resistance to half the moon, like deep sandy loam, adapt to Plain environment.The annual increment of Seedling Stage is less, arrives
Vigorous growth stage, resistance to pruning just can be entered after closing.Growing height and fluffy footpath are all up to 1.2m.Originate in Japanese, China Beijing,
All there is cultivation on the ground such as Nanjing, Qingdao, Mount Lushan, Shanghai, Hangzhou, in the new and better species kind register that Shanghai City greening management board is recommended
Just there is this kind of plant.Ornamental value is good, can be used for multiple occasion, is the Cupressaceae plant of the most rare color stable.Due to it
Reproduction speed is slow, and market is not the most sold.
Sa Wanabai is for newly introducing Colored-leaf Plants, and this plant can carry out sowing, cuttage and mound layering, but all deposits
Deficiency certain: easily make a variation with seminal propagation, is unfavorable for the holding of hereditary character；The cottage propagation life-span is short, root system is weak shallow,
Growth potential is weak, and easily phenomenon is preced with in generation partially, the few factor being also restriction Sa Wanabai and developing in fringe bar source；Mound layering because of
Disposable reproductive number can not carry out large-scale production less.
At present, there is Sa Wanabai just for the research of method for tissue culture, but still suffered from some defects, such as inductivity
Low, more seriously proliferation times and propagation algebraic step, it is impossible to meet requirement.
Summary of the invention
Goal of the invention: in order to solve above-mentioned technical problem, the present invention provides a kind of Sa Wanabai subculture method.
Technical scheme: in order to realize foregoing invention purpose, the invention discloses a kind of Sa Wanabai subculture method, bag
Include following steps:
(1) preparation of culture medium: select basal medium, adds sucrose and agar, add 6-benzyl aminoadenine and
Naphthalene acetic acid, regulates pH value, sterilizing, to obtain final product；
(2) outer implant processes: the Sa Wanabai withered and yellow part of stem section in Primary culture cut off, and cleans；
(3) inoculated and cultured: outer for step (2) the gained old wound of implant is cut off, in inserting step (1) gained culture medium, puts
In cultivating indoor cultivation.
As preferably, in described step (1), basal medium is MS, 1/2MS, B5 or WPM culture medium.
Preferred as another kind, in described step (1), the concentration of sucrose and agar is respectively 30g/L and 7g/L.
Preferred as another kind, in described step (1), the concentration of 6-benzyl aminoadenine is 0.4-0.6mg/L, described naphthalene
The concentration of acetic acid is 0.04-0.06mg/L.
Preferred as another kind, in described step (1), pH value is 5.8～6.0, and sterilising conditions is: sterilizing under 121 DEG C of high pressure
25min。
Preferred as another kind, in described step (2), cleaning method is: first with detergent immersion clean after, then use pure water
Rinse 2 hours.
Preferred as another kind, in described step (3), the condition of culture of culturing room is: illumination 16h/d, and illuminance is
2000lx, temperature controls in (25 ± 2) DEG C
Technique effect: relative to prior art, the inventive method not only has the advantage of tissue culture propagation in prior art, also
By specific culture medium, hormone kind and concentration, combine with other processing method, it is possible to (1) significantly improve successive transfer culture
Proliferation times reach more than 2 times；(2) increase the propagation algebraically of successive transfer culture, make originally can to breed generation can reach 10 generations with
On, improve about more than 100%；(3) scale of Sa Wanabai, factorial praluction is made to be possibly realized.
Detailed description of the invention
Embodiment 1
(1) preparation of culture medium: select culture medium based on MS culture medium, adds sucrose and agar, and concentration is respectively
30g/L and 7g/L, adds 6-benzyl aminoadenine and naphthalene acetic acid, and its concentration is respectively 0.4mg/L and 0.04mg/L, regulates pH
Value is 5.8～6.0, sterilizing 25min under 121 DEG C of high pressure, to obtain final product；
(2) outer implant processes: the Sa Wanabai withered and yellow part of stem section in Primary culture cut off, and first does with detergent immersion
After Jing, then by pure water rinsing 2 hours；
(3) inoculated and cultured: outer for step (2) the gained old wound of implant is cut off, in inserting step (1) gained culture medium, puts
In cultivating indoor cultivation, condition of culture is: illumination 16h/d, and illuminance is 2000lx, and temperature controls in (25 ± 2) DEG C.
Embodiment 2:
(1) preparation of culture medium: select culture medium based on B5 medium, adds sucrose and agar, and concentration is respectively
30g/L and 7g/L, adds 6-benzyl aminoadenine and naphthalene acetic acid, and its concentration is respectively 0.6mg/L and 0.06mg/L, regulates pH
Value is 5.8～6.0, sterilizing 25min under 121 DEG C of high pressure, to obtain final product；
(2) outer implant processes: the Sa Wanabai withered and yellow part of stem section in Primary culture is cut off, first uses detergent immersion
After Gan Jing, then by pure water rinsing 2 hours；
(3) inoculated and cultured: outer for step (2) the gained old wound of implant is cut off, in inserting step (1) gained culture medium, puts
In cultivating indoor cultivation, condition of culture is: illumination 16h/d, and illuminance is 2000lx, and temperature controls in (25 ± 2) DEG C.
Embodiment 3:
(1) preparation of culture medium: select culture medium based on 1/2MS culture medium, adds sucrose and agar, and concentration is respectively
For 30g/L and 7g/L, then adding 6-benzyl aminoadenine and naphthalene acetic acid, its concentration is respectively 0.5mg/L and 0.05mg/L, regulates pH
Value is 5.8～6.0, sterilizing 25min under 121 DEG C of high pressure, to obtain final product；
(2) outer implant processes: the Sa Wanabai withered and yellow part of stem section in Primary culture cut off, and first does with detergent immersion
After Jing, then by pure water rinsing 2 hours；
(3) inoculated and cultured: outer for step (2) the gained old wound of implant is cut off, in inserting step (1) gained culture medium, puts
In cultivating indoor cultivation, condition of culture is: illumination 16h/d, and illuminance is 2000lx, and temperature controls in (25 ± 2) DEG C.
Embodiment 4:
(1) preparation of culture medium: select culture medium based on 1/2MS culture medium, adds sucrose and agar, and concentration is respectively
For 30g/L and 7g/L, adding 6-benzyl aminoadenine and naphthalene acetic acid, its concentration is respectively 0.4mg/L and 0.05mg/L, regulation
PH value is 5.8～6.0, sterilizing 25min under 121 DEG C of high pressure, to obtain final product；
(2) outer implant processes: the Sa Wanabai withered and yellow part of stem section in Primary culture cut off, and first does with detergent immersion
After Jing, then by pure water rinsing 2 hours；
(3) inoculated and cultured: outer for step (2) the gained old wound of implant is cut off, in inserting step (1) gained culture medium, puts
In cultivating indoor cultivation, condition of culture is: illumination 16h/d, and illuminance is 2000lx, and temperature controls in (25 ± 2) DEG C.
Embodiment 5:
(1) preparation of culture medium: select culture medium based on 1/2MS culture medium, adds sucrose and agar, and concentration is respectively
For 30g/L and 7g/L, adding 6-benzyl aminoadenine and naphthalene acetic acid, its concentration is respectively 0.5mg/L and 0.06mg/L, regulation
PH value is 5.8～6.0, sterilizing 25min under 121 DEG C of high pressure, to obtain final product；
(2) outer implant processes: the Sa Wanabai withered and yellow part of stem section in Primary culture cut off, and first does with detergent immersion
After Jing, then by pure water rinsing 2 hours；
(3) inoculated and cultured: outer for step (2) the gained old wound of implant is cut off, in inserting step (1) gained culture medium, puts
In cultivating indoor cultivation, condition of culture is: illumination 16h/d, and illuminance is 2000lx, and temperature controls in (25 ± 2) DEG C.
Experimental example
Take and carry out successive transfer culture with a collection of Sa Wanabai, be set to compare 1 group, compare 2 groups, embodiment 3,4 and 5 groups；
Compare 1 group, for according to the embodiment of the present invention 3 method, remove 6-benzyl aminoadenine income approach and cultivate；
Compare 2 groups, for according to the embodiment of the present invention 3 method, remove naphthalene acetic acid income approach and cultivate；
Embodiment 3,4 and 5 groups is cultivated according to the embodiment of the present invention 3,4 and 5 method respectively；
Each group incubation time is 1 month, investigates final inductivity, proliferation times and increment algebraically, see table 1
Table 1 is respectively organized cultivation results and is investigated
Can be obtained by upper table 1 result, compare comparison 1 group and comparison 2 groups, subculture method of the present invention, it is possible to notable (1)
The proliferation times improving successive transfer culture reaches more than 2 times；(2) increase the propagation algebraically of successive transfer culture, make originally can breed generation
Can reach 10 generations more than, improve about more than 100%；(3) scale of Sa Wanabai, factorial praluction is made to be possibly realized.
Claims (7)
1. Yi Zhong Sa Wanabai subculture method, it is characterised in that comprise the following steps:
(1) preparation of culture medium: select basal medium, adds sucrose and agar, adds 6-benzyl aminoadenine and naphthalene second
Acid, regulates pH value, sterilizing, to obtain final product；
(2) outer implant processes: the Sa Wanabai withered and yellow part of stem section in Primary culture cut off, and cleans；
(3) inoculated and cultured: outer for step (2) the gained old wound of implant is cut off, in inserting step (1) gained culture medium, is placed in training
Support indoor cultivation.
Sa Wanabai subculture method the most according to claim 1, it is characterised in that basis training in described step (1)
Foster base is MS, 1/2MS, B5 or WPM culture medium.
Sa Wanabai subculture method the most according to claim 1, it is characterised in that in described step (1) sucrose and
The concentration of agar is respectively 30g/L and 7g/L.
Sa Wanabai subculture method the most according to claim 1, it is characterised in that 6-benzyl ammonia in described step (1)
The concentration of base adenine is 0.4-0.6mg/L, and the concentration of described naphthalene acetic acid is 0.04-0.06mg/L.
Sa Wanabai subculture method the most according to claim 1, it is characterised in that in described step (1), pH value is
5.8～6.0, sterilising conditions is: sterilizing 25min under 121 DEG C of high pressure.
Sa Wanabai subculture method the most according to claim 1, it is characterised in that cleaning side in described step (2)
Method is: first with detergent immersion clean after, then by pure water rinsing 2 hours.
Sa Wanabai subculture method the most according to claim 1, it is characterised in that culturing room in described step (3)
Condition of culture be: illumination 16h/d, illuminance is 2000lx, and temperature controls in (25 ± 2) DEG C.
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| Application Number | Priority Date | Filing Date | Title |
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| CN201610569338.8A CN106171994A (en) | 2016-07-18 | 2016-07-18 | A kind of Sa Wanabai subculture method |
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| CN201610569338.8A CN106171994A (en) | 2016-07-18 | 2016-07-18 | A kind of Sa Wanabai subculture method |
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| CN106171994A true CN106171994A (en) | 2016-12-07 |
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| CN201610569338.8A Pending CN106171994A (en) | 2016-07-18 | 2016-07-18 | A kind of Sa Wanabai subculture method |
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN106613979A (en) * | 2016-12-23 | 2017-05-10 | 江苏农林职业技术学院 | Primary culture method of cupressus macrocarpa goldcres |
| CN113016617A (en) * | 2021-04-14 | 2021-06-25 | 江苏绿冶苗木科技有限公司 | Tissue culture rapid propagation method of blue lake cypress |
Citations (4)
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|---|---|---|---|---|
| CN1806513A (en) * | 2006-01-24 | 2006-07-26 | 王录顺 | Cold-resistant cypress breeding method |
| JP2010142145A (en) * | 2008-12-17 | 2010-07-01 | Japan Agribio Co Ltd | Somatic embryo mass dividing method |
| CN104871981A (en) * | 2015-07-01 | 2015-09-02 | 临沂大学 | Western red cedar tissue culture rapid propagation method |
| CN105104201A (en) * | 2015-09-09 | 2015-12-02 | 江苏农林职业技术学院 | Chamaecyparis pisifera primary tissue culture method |
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2016
- 2016-07-18 CN CN201610569338.8A patent/CN106171994A/en active Pending
Patent Citations (4)
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| CN1806513A (en) * | 2006-01-24 | 2006-07-26 | 王录顺 | Cold-resistant cypress breeding method |
| JP2010142145A (en) * | 2008-12-17 | 2010-07-01 | Japan Agribio Co Ltd | Somatic embryo mass dividing method |
| CN104871981A (en) * | 2015-07-01 | 2015-09-02 | 临沂大学 | Western red cedar tissue culture rapid propagation method |
| CN105104201A (en) * | 2015-09-09 | 2015-12-02 | 江苏农林职业技术学院 | Chamaecyparis pisifera primary tissue culture method |
Non-Patent Citations (4)
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| 王洪习、蔡冬元: "《植物组织培养技术》", 31 December 2013, 机械工业出版社 * |
| 袁云香: "《金属元素与植物组织培养》", 31 July 2013, 西北农林科技大学出版社 * |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN106613979A (en) * | 2016-12-23 | 2017-05-10 | 江苏农林职业技术学院 | Primary culture method of cupressus macrocarpa goldcres |
| CN113016617A (en) * | 2021-04-14 | 2021-06-25 | 江苏绿冶苗木科技有限公司 | Tissue culture rapid propagation method of blue lake cypress |
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Application publication date: 20161207 |