CN102349438A - Tissue culture and rapid propagation breeding method for golden elf sundust - Google Patents
Tissue culture and rapid propagation breeding method for golden elf sundust Download PDFInfo
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- CN102349438A CN102349438A CN2011102575480A CN201110257548A CN102349438A CN 102349438 A CN102349438 A CN 102349438A CN 2011102575480 A CN2011102575480 A CN 2011102575480A CN 201110257548 A CN201110257548 A CN 201110257548A CN 102349438 A CN102349438 A CN 102349438A
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Abstract
The invention discloses a tissue culture and rapid propagation breeding method for golden elf sundust. The tissue culture and rapid propagation breeding method comprises the following steps of: with lateral bud and stem tip as primary culture materials, soaking the primary culture materials with a sanmate solution; soaking the mixed solution with a vitamin C distilled water solution; and conventionally sterilizing with 0.1 percent liter by percent of mercury, wherein the culture conditions for primary induction are realized by 1/2MS+sucrose+agar+activated carbon+sodium sulfite+TDZ (Thidiazuron)+NAA(Naphthaleneacetic Acid); the culture conditions for subculture multiplication are realized by 1/2MS+agar+sucrose+activated carbon+sodium sulfite+TDZ+NAA; culture conditions for rooting are realized by 1/2MS+agar+sucrose+activated carbon+sodium sulfite+6-BA(Butyl Acrylate)+NAA; and the transplanting conditions for test-tube plantlets are realized by sterilizing seedlings with aqueous solution of potassium permanganate and transplanting the sterilized seedlings. The invention provides a tissue culture method for the golden elf sundust which can be industrially cultured on a large scale.
Description
Technical field
The present invention relates to a kind of sword-leaved cymbidium hybrid new breed gold prodigy's tissue-culturing rapid propagation seedling-cultivating method.
Background technology
The gold prodigy (
GoldenElf ' Sundust ') be that China Taiwan Province is blue with Cymbidium hookerianum hybridization and get with the four seasons before more than 10 year, bloom phenology and proterties more approach the growth characteristics of sword-leaved cymbidium, so generally it are classified as the sword-leaved cymbidium kind at present.It has pattern golden yellow plain, the fragrance of a flower is just pure; Characteristics such as leaf appearance unrestrainedness; Be subjected to consumer's favor since entering the market deeply because of its " inexpensive " always; The general trend of market development is fabulous; Existing 2 swords/basin is bloomed the plant price about 160 yuan/basin; Less to the research of ' gold prodigy ' at present, group is cultivated the research of seedling technical elements and is not appeared in the newspapers as yet.
Sword-leaved cymbidium (
Cymbidium ensifolium) delicate fragrance attacks the people, pattern is plain, cultivation history is long, and is various in style, mainly is distributed in all provinces in China western part and the southeast, distribute with Fujian especially and cultivation the widest, be that China outlet state the earliest is one of blue.China's sword-leaved cymbidium group is cultivated seedling technical research starting early; The history of existing three more than ten years; But because the sword-leaved cymbidium genetic character is complicated; It is main that tradition is viewed and admired with strange flower, different leaf; Thereby be main with seed hybrid breeding and division propagation mainly in producing, because reproduction coefficient is low, new varieties are in often sky-high price propagation of listing stage just; Thereafter drop fast again brings bad influence to the research of sword-leaved cymbidium scale propagation technique, production.Along with the progress of sword-leaved cymbidium crossbreeding technology and the transformation of consumption concept; Color leaf, fragrant flower, the sword-leaved cymbidium kind of the multiple characters combination such as spending more progressively become the new lover in market; Like gold prodigy, line skill sword-leaved cymbidium, color heart sword-leaved cymbidium etc.; Make these kinds enter into common people house; Have only group to cultivate the unique channel that the seedling technology is only its merit of assurance, but the group of at present relevant sword-leaved cymbidium training research is also reported very few.
Summary of the invention
The purpose of this invention is to provide a kind of ability and implement the gold prodigy's of large-scale industrialized cultivation method for tissue culture.
Gold prodigy's of the present invention tissue-culturing rapid propagation seedling-cultivating method comprises the anti-brownization processing of outer plant corpus sterilization, first inducing culture, shoot proliferation cultivation, culture of rootage and test-tube seedling transplanting, may further comprise the steps:
(1) the anti-brownization processing of explant sterilization: getting gold prodigy lateral bud stem apex is the initial culture material; Older tissue prunes away after the collection; Under running water, rinse well; With 0.1% carbendazim+0.2% detergent aqueous solution soaking 30 minutes; Rinse well; Soaked 30 minutes with anti-brownization agent vitamin C distilled water solution (2%) then,, on superclean bench, separate the lateral bud stem apex at last with 0.1% mercuric chloride routine disinfection 10 minutes;
(2) just the inducing culture condition is: 1/2MS+20-30g/L sucrose+8-10g/L agar+3.0mg/L TDZ+0.2mg/L NAA is a medium; Medium pH 5.8; Illumination cultivation 14 hr/d; Intensity of illumination 1500-2000 Lux; Cultivation temperature is 25 ± 2 ℃;
(3) the shoot proliferation condition of culture is: 1/2MS+10g/L agar+30g/L sucrose+1-7mg/L TDZ+0.05-0.4mg/L NAA is a medium; Medium pH 5.8; Illumination cultivation 14hr/d; Intensity of illumination 1500-2000Lux; Cultivation temperature is 25 ± 2 ℃;
(4) the culture of rootage condition is: 1/2MS+10g/L agar+35g/L sucrose+1g/L active carbon+1g/L sodium sulphite+0.1-0.9mg/L 6-BA+0.4-2.0mg/L NAA is a medium, medium pH 5.8; Illumination cultivation 14hr/d; Intensity of illumination 1500-2000Lux; Cultivation temperature is 25 ± 2 ℃;
(5) the test-tube seedling transplanting condition is: culture of rootage was refined seedling 20 days after 60 days again; Transplant after disinfecting seedling with 0.05% potassium permanganate solution; Room temperature is controlled at 15-30 ℃, and air humidity shelters from heat or light 60 ~ 70% at 80%-90%.
TDZ is 3-3.5mg/L in the gold prodigy's of the present invention tissue-culturing rapid propagation seedling-cultivating method shoot proliferation medium, and NAA is 0.1-0.2mg/L.
Gold prodigy's of the present invention tissue-culturing rapid propagation seedling-cultivating method is characterized in that: the 6-BA in the root media is 0.3-0.5mg/L, and NAA is 0.8-1.2mg/L.
Gold prodigy's of the present invention tissue-culturing rapid propagation seedling-cultivating method is characterized in that: can add 1g/L active carbon and 1g/L sodium sulphite in first inducing culture and the shoot proliferation medium.
Gold prodigy's of the present invention tissue-culturing rapid propagation seedling-cultivating method is characterized in that: the matrix of test-tube seedling transplanting is vermiculite or bark.
The beneficial effect that the present invention reaches:
(1) the lateral bud stem apex is used 0.1% mercuric chloride routine disinfection with 2% vitamin C aqueous solution soaking again after 30 minutes before sterilization, can effectively delay brownization of the explant time and keep the explant vigor;
(2) after interpolation resists brownization agent 1g/L active carbon and 1g/L sodium sulphite simultaneously in the 1/2MS medium; Explant, protocorm and the bud of growing thickly produce brownization in can reduction group training process; Reached at present that explant is just induced, brownization of protocorm shoot proliferation rate is controlled at below 15%, brownization of blastogenesis root induction of growing thickly rate is lower than below 5%;
(3) concentration is that the TDZ of 3.0-3.5mg/L and NAA that concentration is 0.1-0.2mg/L combination help explant and just induce with protocorm and breed; Along with the increase of TDZ concentration, help protocorm and break up the bud of growing thickly, otherwise then help protocorm differentiation protocorm, growth coefficient reaches 7.33, and shoot proliferation is cultivated 4-5 generation continuously; Concentration is the 6-BA of 0.3-0.5 mg/L and NAA that concentration is 0.8-1.2 mg/L combination, helps the blastogenesis root induction of growing thickly most, and inductivity reaches 100.0%;
(4) the test-tube seedling transplanting condition is: culture of rootage was refined seedling 20 days after 60 days again; Transplant after disinfecting seedling with 0.05% potassium permanganate solution; Room temperature is controlled at 15-30 ℃, and air humidity is at 80%-90%, shelters from heat or light 70% to transplant, and adopting vermiculite or bark is transplanting medium, and survival rate reaches more than 95%;
(5) initial culture base of the present invention and shoot proliferation medium difference are less, grow seedlings than being easier to industrialized tissue culture.
Embodiment:
Getting gold prodigy 0.4cm lateral bud stem apex is the initial culture explant.With the older tissue that prunes away after the above-mentioned material collection, under running water, rinse well, with 0.1% carbendazim+0.2% detergent aqueous solution soaking 30 minutes, rinse well; Soaked 30 minutes with anti-brownization agent vitamin C distilled water solution (2%) then,, on superclean bench, separate the lateral bud stem apex at last with 0.1% mercuric chloride routine disinfection 10 minutes; Be seeded in just on the inducing culture first inducing culture based formulas: 1/2MS+30g/L sucrose+10g/L agar+1g/L active carbon+1g/L sodium sulphite+3.5mg/L TDZ+0.2mg/L NAA with peeling off lateral bud stem apex after the sterilization; Light application time 14hr/d; Intensity of illumination 1800lux; Cultivation temperature (25 ± 2 ℃); About initial culture 60 days, the lateral bud stem apex is induced and is differentiated 2-3 diameter 0.2cm protocorm, with its cutting and separating, inoculation; With 1/2MS+10g/L agar+30g/L sucrose is minimal medium, adds the 1g/L active carbon and adds 1g/L active carbon+1g/L sodium sulphite; Shoot proliferation was cultivated after 60 days, and protocorm differentiates grow thickly bud and protocorm simultaneously, and the bud of will growing thickly is inoculated after cutting into 2 protocorm simple buds of base portion band; With 1/2MS+10g/L agar+30g/L sucrose+1g/L active carbon+1g/L sodium sulphite+3mg/L TDZ+0.2mg/L NAA is the shoot proliferation medium, and growth coefficient reaches 7.33, and shoot proliferation is cultivated 4-5 generation continuously; The 1-1.5cm height that protocorm is induced is grown thickly and is inoculated after bud is divided into the unrooted simple bud; The root induction medium: 1/2MS+8g/L agar+25g/L sucrose+1g/L active carbon+0.5mg/L 6-BA+1.2mg/L NAA, root induction is after 60 days, 100% root induction, the 2-3 bar of taking root, the long 0.5-3.5cm of root; After the culture of rootage 60 days, refined seedling again 20 days; Use vermiculite or be transplanting medium, transplant after disinfecting seedling with 0.05% potassium permanganate solution, room temperature is controlled at 25-30 ℃, and air humidity shelters from heat or light 70% at 80%-90%, and transplanting survival rate reaches 96%.
Claims (5)
1. gold prodigy's tissue-culturing rapid propagation seedling-cultivating method comprises the anti-brownization processing of explant sterilization, first inducing culture, shoot proliferation cultivation, culture of rootage and test-tube seedling transplanting, it is characterized in that:
(1) the anti-brownization processing of explant sterilization: getting gold prodigy lateral bud stem apex is the initial culture material; Older tissue prunes away after the collection; Under running water, rinse well; With 0.1% carbendazim+0.2% detergent aqueous solution soaking 30 minutes; Rinse well; Soaked 30 minutes with anti-brownization agent vitamin C distilled water solution (2%) then,, on superclean bench, separate the lateral bud stem apex at last with 0.1% mercuric chloride routine disinfection 10 minutes;
(2) first inducing culture, medium is: 1/2MS+20-30g/L sucrose+8-10g/L agar+3.0-3.5mg/L TDZ+0.1-0.2mg/L NAA, pH 5.8, illumination cultivation 14 hr/d, intensity of illumination 1500-2000 Lux, cultivation temperature is 25 ± 2 ℃;
(3) shoot proliferation is cultivated, and medium is: 1/2MS+10g/L agar+30g/L sucrose+1-7mg/L TDZ+0.05-0.4mg/L NAA, and medium pH 5.8, illumination cultivation 14hr/d, intensity of illumination 1500-2000Lux, cultivation temperature is 25 ± 2 ℃;
(4) culture of rootage; Medium is: 1/2MS+10g/L agar+35g/L sucrose+1g/L active carbon+1g/L sodium sulphite+0.1-0.9mg/L 6-BA+0.4-2.0mg/L NAA, and pH 5.8, illumination cultivation 14hr/d; Intensity of illumination 1500-2000Lux, cultivation temperature is 25 ± 2 ℃;
(5) the test-tube seedling transplanting condition is: culture of rootage was refined seedling 20 days after 60 days again, transplanted after disinfecting seedling with 0.05% potassium permanganate solution, and room temperature 15-30 ℃, air humidity 80%-90% shelters from heat or light 60 ~ 70%.
2. gold prodigy's according to claim 1 tissue-culturing rapid propagation seedling-cultivating method is characterized in that: TDZ is 3-3.5mg/L in the shoot proliferation medium, and NAA is 0.1-0.2mg/L.
3. gold prodigy's according to claim 1 tissue-culturing rapid propagation seedling-cultivating method is characterized in that: the 6-BA in the root media is 0.3-0.5mg/L, and NAA is 0.8-1.2mg/L.
4. gold prodigy's according to claim 1 tissue-culturing rapid propagation seedling-cultivating method is characterized in that: just induce with the shoot proliferation medium in can add 1g/L active carbon and 1g/L sodium sulphite.
5. gold prodigy's according to claim 1 tissue-culturing rapid propagation seedling-cultivating method is characterized in that: the matrix of test-tube seedling transplanting is vermiculite or bark.
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Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103416303A (en) * | 2013-07-10 | 2013-12-04 | 浙江建设职业技术学院 | Inducing method for spring dendrobium protoemm-like body |
| CN104521751A (en) * | 2014-12-04 | 2015-04-22 | 中国科学院华南植物园 | Tissue culture efficient breeding and seedling growing method of hybrid orchid Huangjinxiaoshentong |
| CN106718885A (en) * | 2016-11-29 | 2017-05-31 | 广西南岜仔科技有限公司 | A kind of culture medium of efficient rapid induction plum blossom Callus formation |
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|---|---|---|---|---|
| CN1247022A (en) * | 1999-08-20 | 2000-03-15 | 华南师范大学 | Quickly in-vitro breeding method of macranthus Hui orchid |
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| Publication number | Priority date | Publication date | Assignee | Title |
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| CN1247022A (en) * | 1999-08-20 | 2000-03-15 | 华南师范大学 | Quickly in-vitro breeding method of macranthus Hui orchid |
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| Title |
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| 《浙江农业科学》 20081231 李丽等 线艺兰的组织培养及植株再生研究 1-5 , 第6期 * |
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103416303A (en) * | 2013-07-10 | 2013-12-04 | 浙江建设职业技术学院 | Inducing method for spring dendrobium protoemm-like body |
| CN104521751A (en) * | 2014-12-04 | 2015-04-22 | 中国科学院华南植物园 | Tissue culture efficient breeding and seedling growing method of hybrid orchid Huangjinxiaoshentong |
| CN106718885A (en) * | 2016-11-29 | 2017-05-31 | 广西南岜仔科技有限公司 | A kind of culture medium of efficient rapid induction plum blossom Callus formation |
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Application publication date: 20120215 Assignee: QIANXINAN GREEN ANIMAL AND PLANT SCIENCE AND TECHNOLOGY DEVELOPMENT CO.,LTD. Assignor: GUIZHOU INSTITUTE OF BIOLOGY Contract record no.: 2015520000010 Denomination of invention: Tissue culture and rapid propagation breeding method for golden elf sundust Granted publication date: 20130313 License type: Exclusive License Record date: 20150527 |
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