CN1586166A - Quick breeding method for cymbidium hyridus high quality sprout - Google Patents
Quick breeding method for cymbidium hyridus high quality sprout Download PDFInfo
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- CN1586166A CN1586166A CN 200410050907 CN200410050907A CN1586166A CN 1586166 A CN1586166 A CN 1586166A CN 200410050907 CN200410050907 CN 200410050907 CN 200410050907 A CN200410050907 A CN 200410050907A CN 1586166 A CN1586166 A CN 1586166A
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Abstract
The present invention relates to the quick breeding method of high quality seedling of Cymbidium hyridus, as one hybrid of orchid and the culture medium therewith. The present invention provides improved culture medium component, technological process, etc. and has method with high successive dirt-removing rate, high proliferation rate, low variation rate, robust seedling, high quality, high survival rate and other advantages. In addition, the present invention has high sterilization efficiency, unique culture medium component, low cost and high application value.
Description
Technical field
The invention provides hybrid cymbidium high quality seedling method for quickly breeding and employed medium.
Background technology
Hybrid cymbidium is by Da Hua class original seed hybridization in Cymbidium (cymbidium) plant.Domestic in early days after abroad introduce, because of it spends number numerous, large flower and brilliant color, plant is tall and big, so be called hybrid cymbidium, the hybrid cymbidium kind reaches more than 10,000 at present, this year by with Cymbidium in sword-leaved cymbidium, the kind that much has pleasant fragrance has also been cultivated in fragrant flower inter breed crossings such as Chunlan.Hybrid cymbidium forms the spending spree in recent years at home, the annual need from Japan, a large amount of commodity plant of Korea S's import, and domestic commodity production development is rapid, and seedling is in great shortage at present.The production of hybrid cymbidium commercial seedling is almost all bred with test-tube plantlet, adopts tissue culture technology to produce high-quality test-tube plantlet, for the needs that satisfy seedling market provide a valid approach.The tissue culture technique research of hybrid cymbidium is carried out early abroad, is that first carries out the kind that the orchid batch production is produced.Domesticly also carried out relevant research in recent years, had the minority group training company to have carried out large-scale production and the report of a routine patent application is arranged.
Summary of the invention
The tissue culture propagation and the employed medium that the purpose of this invention is to provide hybrid cymbidium.Just in generation, carried out system research with many for the tissue culture and the quick propagating technology of hybridizing the hybrid cymbidium kind of cultivating to several in the present invention, from the angle of producing to medium component, there is bigger innovation aspects such as technological process, and a cover hybrid cymbidium plant high quality seedling method for quickly breeding and relevant culture medium prescription is provided.Its explant disinfection efficiency height, medium component uniqueness and cost are low, the seedling first-class quality of producing, and the using value height specifically comprises the steps:
1. material: the present invention adopts material that the hybrid cymbidium of 6 kinds is arranged: green pearl, pianist, golden yellow prodigy, Taibei Miss, ivory Cymbidium hookerianum (cymbidium tracyanum * cymbidium ebureum).
2. the method for explant induction materials disinfection: cut the hybrid cymbidium sprouting, flowing water flushing 5 minutes, with explant on the superclean bench with alcohol-pickled 30 seconds after, soaked 10 minutes with weight percent concentration 0.1% mercuric chloride again, aseptic water washing 3 times, divest the bud bract with scalpel, put into the mercuric chloride solution 5 minutes of weight percent concentration 0.1% again, sterile water is given a baby a bath on the third day after its birth time, successively peels off blade, every stripping one deck blade sheet, soaked 1 minute with weight percent concentration 0.1% mercuric chloride, aseptic water washing three times is until shelling to exposing growing point.
3, intending protocorm induces: cut 2-5mm
3About the piece of tissue of band growing point be inoculated into and intend the protocorm inducing culture: MH+6-benzyl purine 2-10 mg/litre+methyl 0.2-1.0 mg/litre+10-20% Sucus Cocois+citric acid 0-100 grams per liter.Sucus Cocois by the juice in the coconut of fresh listing after filtration.Add coconut milk and can obviously improve the inductivity of intending protocorm.
4, shoot proliferation: cultivate and to intend protocorm in about 30 days and transfer to the shoot proliferation medium: MH+6-benzyl purine 0.2-0.5 mg/litre+methyl 0.2-0.5 mg/litre+active carbon 0.5 gram/l.Enrichment culture was a subculture cycle in general about 30 days.Removing coconut milk in breeding mainly is in order to reduce cost, to adopt the hormone combinations of low concentration and interpolation small amount of activated can reduce brownization, reduce the generation of variation under the condition of the certain rate of increase of maintenance.
5, become seedling to cultivate: to intend protocorm and breed to be inoculated into into behind certain algebraically and seedling medium culture 2-3 week form seedling.Become the seedling medium: HF1+6-benzyl purine 0.5-1 mg/litre+methyl 0.1-0.2 mg/litre.
6. strong plantlets and rootage is cultivated: when 2-3 centimetre of unrooted height of seedling, be transferred on the strong plantlets and rootage medium and cultivate, add the active carbon radical and increase, can prevent the portions cut brown stain. strong plantlets and rootage medium: HF2+ methyl 0.5-1 mg/litre+caseinhydrolysate 0.2 grams per liter+banana homogenate 10%+0.5% active carbon.
Above medium all contains sucrose 20 grams per liters, pH5.2-5.4, agar 0.7%.Cultivation temperature (28 ± 2) ℃, illuminance 1500～2000lx, illumination 12 hours/day.
7. test-tube seedling transplanting: when strong seedling culture about 6 weeks, test-tube plantlet can grow to 10-12 centimetre high, transfer to the natural daylight lower refining seedling after 7 days, it is taken out from vial, clean the medium of root, move into bark: pool foundation stone is in 2: 1 the mixed-matrix, to keep suitably ventilating and enough humidity, 1 all left and right sides test-tube plantlet can recover growth, and the survival rate of transplanting all can reach 95%-100%.
Carrying out hardening handles and to help the cuticular formation of test-tube plantlet and to the adaptive capacity of environment, transplanting survival rate increases.
It is higher that method provided by the present invention has explant decontamination success rate, rate of increase height, advantage such as aberration rate is low, and seedling is sturdy, quality better, transplants back survival rate height, and growth potential is strong.Its explant disinfection efficiency height, medium component uniqueness and cost are low, the seedling first-class quality of producing, using value height.
Technical characterstic:
1. the hybrid cymbidium commodity production all adopts test-tube plantlet to breed, and explant decontamination difficulty when carrying out tissue culture, it is explant and multistep disinfectant method that this patent adopts the sprouting of not leafing, the effect of sterilization can reach 80% success rate;
2. when protocorm was bred, growth coefficient and variation frequency direct ratio often increased, and this patent adopts low hormone concentration and adds active carbon in proliferated culture medium, had reached the higher rate of increase and the effect that prevents to make a variation;
3. the cultivation in the product of seedling confrontation later stage has great influence in the seedling production of hybrid cymbidium, the commodity hybrid cymbidium quality of existing domestic production is difficult to and the comparing of Japanese import, except that the culture technique influence, the seedling poor quality also is a major reason, this patent adopts two step seedling methods, when intending the protocorm subculture, at first strict control do not emerge, help propagation and operation, change over to then on the special one-tenth seedling medium and cultivate, help inducing stalwartness, the hybrid cymbidium seedling of quality homogeneous, further change the strong seedling culture base that adds organic additive over to, seedling was grown up rapidly, can grow the bottle outlet specification about 6 weeks, through 7-14 days hardening, bottle outlet seedling color is dark green, plant height 10-14cm (different) according to kind, radical can reach the 3-6 bar, and shoot root is more sturdy with general seedling, seedling and root length ratio are about 3: 2, are easy to transplant.
4. the seedling cultivated of this patent has reached very high quality, spends precious complex medium and adopt in this step, and is easy to use, cost is low, effect is remarkable.
Embodiment
Embodiment 1
1. draw materials: the ivory Cymbidium hookerianum is the sprouting of leafing not.
2. the method for explant induction materials disinfection: cut the ivory Cymbidium hookerianum kind sprouting of cultivating in the greenhouse with sharp blade from base portion, running water flushing 5 minutes.With explant on the superclean bench with alcohol-pickled 30 seconds after, soaked 10 minutes aseptic water washing 3 times again with 0.1% mercuric chloride, divest the bud bract with scalpel,, put into 0.1% mercuric chloride solution again 5 minutes, sterile water is given a baby a bath on the third day after its birth time, successively peel off blade, every stripping one deck blade sheet soaked 1 minute aseptic water washing three times with 0.1% mercuric chloride, to exposing growing point, the piece of tissue that cuts the band growing point about 2-5mm3 is inoculated into intends inducing on the protocorm inducing culture plan protocorm stem eye to form until stripping.
3. intending protocorm induces: the piece of tissue that cuts the band growing point about 2-5mm3 is inoculated into intends the protocorm inducing culture: MH+6-benzyl purine 2 mg/litre+methyl 0.2 mg/litre+15% Sucus Cocois+citric acid 50 grams per liters; Cultivating had the protocorm of plan form in about 30 days.
4. shoot proliferation: inducing culture will be intended protocorm and transfer to shoot proliferation on MH+6-benzyl purine 0.2 mg/litre+methyl 0.5 mg/litre+active carbon 0.5 gram/l shoot proliferation medium about 30 days, general about 30 days is a subculture cycle.
5. become seedling to cultivate: to breed and be inoculated into into the seedling medium behind certain algebraically: HF1+6-benzyl purine 0.5 mg/litre+methyl 0.1 mg/litre.The promptly visible plan protocorm in one week back forms shoot apex, can grow up to the unrooted seedling about 3cm 3-4 week.
6. strong plantlets and rootage is cultivated: will become on the seedling medium to cultivate about 4 weeks, the high seedling of about 3cm of formation be transferred to the strong plantlets and rootage medium: HF2+ methyl 1 mg/litre+caseinhydrolysate 0.2 grams per liter+banana homogenate 10%+0.5% active carbon; Every bottle graft kind 22 strains.The plant mean elements is 3.4 after 6 weeks of inoculation, average plant height 9.8cm, and average fresh weight reaches 2.2 gram/l
7. test-tube seedling transplanting: with the bottle seedling in 4 weeks of strong seedling culture, transfer to greenhouse culture condition lower refining seedling 7 days, it is taken out from vial, clean the medium of root, move into bark: pool foundation stone is in 2: 1 the mixed-matrix, keep suitably ventilating and humidity, the survival rate of transplanting all can reach more than 98%.
Above medium all contains sucrose 20 grams per liters, pH5.2-5.4, agar 0.7%.Cultivation temperature (28 ± 2) ℃, illuminance 1500～2000lx, illumination 12 hours/day.Sucus Cocois by the juice in the blue or green coconut after filtration.
Embodiment 2
The sprouting of getting ' pianist ' is an explant.The basic operation method is with embodiment 1 unanimity.But it intends the protocorm inducing culture is MH+6-benzyl purine 5 mg/litre+methyl 0.5 mg/litre+10% Sucus Cocois.Intending the protocorm proliferated culture medium is MH+6-benzyl purine 1 mg/litre+methyl 0.2 mg/litre+active carbon 0.5 gram/l; Becoming the seedling medium is HF1+6-benzyl purine 0.5 mg/litre+methyl 0.2 mg/litre.Strong plantlets and rootage cultivates that the statistical average radical is 4.6 after 40 days, average plant height 10.7cm, and average fresh weight reaches 3.2 grams.The survival rate of test-tube seedling transplanting can reach 100%.
Claims (7)
1, a kind of tissue culture propagation of hybrid cymbidium is characterized in that providing relevant culture medium prescription from the angle of producing to hybrid cymbidium plant high quality seedling method for quickly breeding, and technological process, and concrete steps comprise as follows:
1), the method for explant induction materials disinfection: cut the hybrid cymbidium sprouting, flowing water flushing 5 minutes, with explant on the superclean bench with alcohol-pickled 30 seconds after, soaked 10 minutes with weight percent concentration 0.1% mercuric chloride again, aseptic water washing 3 times, divest the bud bract with scalpel, put into the mercuric chloride solution 5 minutes of weight percent concentration 0.1% again, sterile water is given a baby a bath on the third day after its birth time, successively peels off blade, every stripping one deck blade sheet, soaked 1 minute with weight percent concentration 0.1% mercuric chloride, aseptic water washing three times is until shelling to exposing growing point;
2), intending protocorm induces: cut 2-5mm
3About the piece of tissue of band growing point be inoculated into and intend the protocorm inducing culture;
3), shoot proliferation: cultivate and will intend protocorm in about 30 days and transfer on the shoot proliferation medium, enrichment culture 30 days was a subculture cycle;
4), become seedling to cultivate: to intend protocorm and breed behind certain algebraically and be inoculated into into seedling medium culture 2-3 week, form seedling;
5), strong plantlets and rootage cultivates: when 2-3 centimetre of unrooted height of seedling, be transferred on the strong plantlets and rootage medium and cultivate;
6), test-tube seedling transplanting: when 6 weeks of strong seedling culture, test-tube plantlet can grow to 10-12 centimetre high, transfer to the natural daylight lower refining seedling after 7 days, it is taken out from vial, clean the medium of root, move into bark: pool foundation stone is in 2: 1 the mixed-matrix, to keep suitably ventilating and enough humidity, and 1 all test-tube plantlets can recover to grow.
2, according to the tissue culture propagation of the hybrid cymbidium described in the claim 1, it is characterized in that 2) in intend the protocorm inducing culture and be: MH+6-benzyl purine 2-10 mg/litre+methyl 0.2-1.0 mg/litre+10-20% Sucus Cocois+citric acid 0-100 grams per liter.
3,, it is characterized in that described Sucus Cocois by the juice in the fresh coconut after filtration according to the tissue culture propagation of the hybrid cymbidium described in claim 1 or 2.
4, according to the tissue culture propagation of the hybrid cymbidium described in the claim 1, it is characterized in that 3) in, the shoot proliferation medium, be: MH+6-benzyl purine 0.2-0.5 mg/litre+methyl 0.2-0.5 mg/litre+active carbon 0.5 gram/l.
5, according to the tissue culture propagation of the hybrid cymbidium described in the claim 1, it is characterized in that 4), middle one-tenth seedling medium is: HF1+6-benzyl purine 0.5-1 mg/litre+methyl 0.1-0.2 mg/litre.
6, according to the tissue culture propagation of the hybrid cymbidium described in the claim 1, it is characterized in that 5), middle strong plantlets and rootage medium is: HF2+ methyl 0.5-1 mg/litre+caseinhydrolysate 0.2 grams per liter+banana homogenate 10%+0.5% active carbon.
7, according to the tissue culture propagation of the hybrid cymbidium described in the claim 1, it is characterized in that above plan protocorm inducing culture, shoot proliferation medium, become seedling medium, strong plantlets and rootage medium, all contain sucrose 20 grams per liters, pH5.2-5.4, agar 0.7%; 28 ± 2 ℃ of cultivation temperature, illuminance 1500～2000lx, illumination 12 hours/day.
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Cited By (11)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101889550A (en) * | 2010-08-04 | 2010-11-24 | 扬州大学 | Culture medium and method for culturing tissues of Chinese cymbidium faberi rolfe |
| CN102090252A (en) * | 2010-11-10 | 2011-06-15 | 天津滨海国际花卉科技园区股份有限公司 | Cuttage propagation method of allium giganteum |
| CN101584298B (en) * | 2009-06-02 | 2011-09-14 | 中国科学院华南植物园 | Test-tube breeding method for Nothodoritis germchit |
| CN102283127A (en) * | 2011-07-21 | 2011-12-21 | 福建省亚热带植物研究所 | Culture medium synchronously used for induction and multiplication of Cymbidium hybridum stem tip Caespitose shoots and use method thereof |
| CN102301950A (en) * | 2011-06-29 | 2012-01-04 | 南开大学 | Industrial production method of Cymbidium germchit based on sexual hybridization |
| CN102754599A (en) * | 2012-07-16 | 2012-10-31 | 上海市农业科学院 | Method for quickly breeding cymbidium hybridium by use of root inducing protocorm |
| CN102893871A (en) * | 2012-10-24 | 2013-01-30 | 泰安市泰山林业科学研究院 | Method for crossbreeding Cymbidium faberi |
| CN103755461A (en) * | 2013-12-23 | 2014-04-30 | 佛山市顺德区今日景艺生物科技有限公司 | Culture medium for culturing orchid tissue and culture method thereof |
| CN107750949A (en) * | 2015-07-22 | 2018-03-06 | 浙江传化生物技术有限公司 | The method that hybrid cymbidium seedling is efficiently bred using stem-tip tissue |
| CN109937875A (en) * | 2019-03-12 | 2019-06-28 | 中国科学院华南植物园 | One kind carrying out pocket orchid high quality seedling quick breeding method for tissue culture by leafage bud |
| CN113475396A (en) * | 2021-07-25 | 2021-10-08 | 杭州市农业科学研究院 | Efficient seedling method for hybrid cymbidium hybridum of spring sword and cymbidium hybridum |
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Cited By (16)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN101584298B (en) * | 2009-06-02 | 2011-09-14 | 中国科学院华南植物园 | Test-tube breeding method for Nothodoritis germchit |
| CN101889550A (en) * | 2010-08-04 | 2010-11-24 | 扬州大学 | Culture medium and method for culturing tissues of Chinese cymbidium faberi rolfe |
| CN102090252B (en) * | 2010-11-10 | 2013-02-20 | 天津滨海国际花卉科技园区股份有限公司 | Cuttage propagation method of allium giganteum |
| CN102090252A (en) * | 2010-11-10 | 2011-06-15 | 天津滨海国际花卉科技园区股份有限公司 | Cuttage propagation method of allium giganteum |
| CN102301950A (en) * | 2011-06-29 | 2012-01-04 | 南开大学 | Industrial production method of Cymbidium germchit based on sexual hybridization |
| CN102283127A (en) * | 2011-07-21 | 2011-12-21 | 福建省亚热带植物研究所 | Culture medium synchronously used for induction and multiplication of Cymbidium hybridum stem tip Caespitose shoots and use method thereof |
| CN102754599B (en) * | 2012-07-16 | 2013-09-18 | 上海市农业科学院 | Method for quickly breeding cymbidium hybridium by use of root inducing protocorm |
| CN102754599A (en) * | 2012-07-16 | 2012-10-31 | 上海市农业科学院 | Method for quickly breeding cymbidium hybridium by use of root inducing protocorm |
| CN102893871A (en) * | 2012-10-24 | 2013-01-30 | 泰安市泰山林业科学研究院 | Method for crossbreeding Cymbidium faberi |
| CN102893871B (en) * | 2012-10-24 | 2013-11-20 | 泰安市泰山林业科学研究院 | Method for crossbreeding Cymbidium faberi |
| CN103755461A (en) * | 2013-12-23 | 2014-04-30 | 佛山市顺德区今日景艺生物科技有限公司 | Culture medium for culturing orchid tissue and culture method thereof |
| CN103755461B (en) * | 2013-12-23 | 2015-09-16 | 佛山市顺德区今日景艺生物科技有限公司 | The medium that Orchid Tissue is cultivated and cultural method thereof |
| CN107750949A (en) * | 2015-07-22 | 2018-03-06 | 浙江传化生物技术有限公司 | The method that hybrid cymbidium seedling is efficiently bred using stem-tip tissue |
| CN109937875A (en) * | 2019-03-12 | 2019-06-28 | 中国科学院华南植物园 | One kind carrying out pocket orchid high quality seedling quick breeding method for tissue culture by leafage bud |
| CN109937875B (en) * | 2019-03-12 | 2020-12-01 | 中国科学院华南植物园 | Rapid propagation method for paphiopedilum high-quality seedling tissue culture through leaf clumpy buds |
| CN113475396A (en) * | 2021-07-25 | 2021-10-08 | 杭州市农业科学研究院 | Efficient seedling method for hybrid cymbidium hybridum of spring sword and cymbidium hybridum |
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