CN1631109A - High quality germchit tissure culturing and rapid breeding method of Dendrobium sp. - Google Patents
High quality germchit tissure culturing and rapid breeding method of Dendrobium sp. Download PDFInfo
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- CN1631109A CN1631109A CN 200410077729 CN200410077729A CN1631109A CN 1631109 A CN1631109 A CN 1631109A CN 200410077729 CN200410077729 CN 200410077729 CN 200410077729 A CN200410077729 A CN 200410077729A CN 1631109 A CN1631109 A CN 1631109A
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Abstract
Disclosed is the high quality germchit tissure culturing and rapid breeding method of Dendrobium sp., wherein a large amount of test tube seedlings with stabilized performance are obtained through fine breed tissue culture by adopting the path of clumping buds. The produced Dendrobium sp. has the advantages of high genetic stability, quick emergence of seedlings, strong seedlings, good seedling quality, strong resistance, and fine growth status.
Description
Technical field
The present invention relates to the high quality seedling method for quickly breeding and the employed medium of autumn dendrobium.
Background technology
The autumn dendrobium claims the butterfly stem of noble dendrobium again, is mostly that the tropical initial species butterfly stem of noble dendrobium (Dendrobiumphalaenopsis) to originate in New Guinea is the Hybrid that the parent breeds.New Guinea, Australia are the native places of autumn stem of noble dendrobium class original seed, and what Hybrid cultivation was maximum is main product ground " panda No. 1 " with Thailand, and other more popular kind has " celestial a kind of thick silk ", " Mary ", promise graceful etc.
The autumn stem of noble dendrobium does not have tangible resting stage, and blade can be kept the several years and not come off; Annual autumn, visible inflorescence was extracted out from the top and near the joint of pseudobulb.Scape is long, and every stem has more than ten flower, and pseudobulb can be left the racemosus flower and bloom the several years continuously sometimes, can make cut-flower and also can make potted flower culture.It is divided into two classes to the autumn stem of noble dendrobium according to big wisp of its flower: a class is the Da Hua type autumn stem of noble dendrobium, its flower shape and flower on inflorescence give birth to the mode utmost point as Moth orchid, just lip is much simple than the lip of Moth orchid, as the hybrid butterfly stem of noble dendrobium, can make the usefulness of potted flower or cut-flower; Another kind of is the Xiao Hua type autumn stem of noble dendrobium, and main feature is that sepal is elongated, upward upright, and the warp shape is arranged.Flower is abundant, and the florescence is long especially, the also suitable especially usefulness of making the potted flower cut-flower.
The autumn stem of noble dendrobium occurs relatively early in China flowers market, and the characteristics of this class dendrobium maximum are under the environment of hot and humid and short-day, could form bud, are suitable for the torrid zone and high temperature greenhouse culture.The general florescence in the fall, but usually as long as the condition permission also may bloom in other season.The original seed of the autumn stem of noble dendrobium (Dendrobium) mainly is distributed in the jungle of Oceanian Australia and Papua New Guinea, often grow nonparasitically upon another plant on the big tree of lowland rainforest or on the cliff, their common feature is the florescence to be confined to autumn, but at present, cultivate through artificial hybridization, many pattern differences have appearred, the popular kind that the florescence is open all year round.The area of autumn stem of noble dendrobium cut-flower output maximum is torrid areas such as Thailand, Malaysia and Hawaii now, and China's cultivation amount is also less, and as a kind of novel high-grade cutting flower variety, market demand is bigger.
China mainland has been introduced many autumn dendrobium kinds in recent years, but plant division is mainly leaned in seedling production, and reproduction speed is slow.The autumn dendrobium seedling of the high-quality of having produced, market is very good.At present, the domestic report that yet there are no autumn dendrobium seedling tissue-culturing quick-propagation success does not have the autumn dendrobium seedling patent of breeding fast yet.
Summary of the invention
The invention provides tissue culture propagation and the employed cultivation of multiple autumn dendrobium original seed and hybrid.To the selection of explant, condition of culture and medium component are systematically discussed, the using value height.The tissue culture of carrying out autumn dendrobium improved seeds by the bud approach of growing thickly obtains the stable test-tube plantlet of a large amount of proterties, for the needs that satisfy autumn dendrobium high quality seedling market provide a valid approach.
Autumn dendrobium high quality seedling method for quickly breeding of the present invention may further comprise the steps:
1. material: the material that this invention has been tested has the butterfly stem of noble dendrobium (Dendrobium phalaenepsis), the goat's horn stem of noble dendrobium (Dendrobium stratiotes), panda No. 1 (Dendrobium Ekapol Pandal '), panda No. 2 (Dendrobium Ekapol Panda 2 '), Thailand white (Dendrobium Thailand White), Zhuan Fu (Dendrobium Joam Ku), purple butterfly (Dendrobium Isumi Kiyoshi), paint face (Dendrobium Kelrme) etc., and material is all taken from the South China Botanical Garden.
2. the sterilization of explant: get the tender shoots that the autumn dendrobium is given birth to tool 5-6 joint then, after removing blade, be cut into two sections after rinsing well with running water, on superclean bench with 70% alcohol-pickled 1 minute, again with 0.1% mercuric chloride solution sterilization 10 minutes, behind the aseptic water washing 5 times, blot with aseptic blotting paper, the stem section and the stem apex of clip 2-3 centimetre of band joint are inoculated on protocorm and the inducing clumping bud medium.
3. protocorm and the inducing of bud of growing thickly: inducing culture is on MS+6-benzyl purine 1-5 mg/litre+methyl 0.2-0.5 mg/litre+15% Sucus Cocois medium, under cultivation temperature (26 ± 2) ℃ condition, left and right sides stem apex began growth in 10 days, there is sprouting to form on 20 days left and right sides stem Duan Jie and growth, when the 6-benzyl purine is that the stylish bastem of 4-5 portion has protocorm to produce.Medium contains sucrose 30 grams per liters, pH 5.5-5.8, agar 0.7%.Illuminance 1500～2000lx, illumination 12 hours/day.
4. adventitious buds proliferation: the bud of growing thickly that will utilize stem section and Shoot Tip Culture to obtain, cut off partial blade and transfer to the adventitious buds proliferation medium: MS+6-benzyl purine 1-5 mg/litre+methyl 0.2-0.5 mg/litre+15% Sucus Cocois, can induce protocorm and grow thickly bud formation; Protocorm can be bred on higher 6-benzyl purine concentration medium, and protocorm can differentiate the bud of growing thickly when changing the medium that hangs down 6-benzyl purine concentration over to, and the bud of growing thickly can be bred on the medium of low 6-benzyl purine concentration.Illuminance 1500～2000lx, illumination 12 hours/day.
5. strong plantlets and rootage is cultivated: adventitious buds proliferation is after many generations, the bud of growing thickly can be cut into single bud, be seeded in and carry out strong plantlets and rootage on the strong plantlets and rootage medium of 1/2MS medium+methyl 0.5-1.0 mg/litre+bananas juice 100 grams per liters+activated carbon 0.5 grams per liter and cultivate.Can form plant height 3-4 centimetre whole plant about 40 days.Illuminance 1500～2000lx, illumination 12 hours/day.
6. test-tube seedling transplanting: spring the favorable season for transplanting.During test-tube seedling transplanting, natural lighting lower refining seedling 5-10 days, autumn dendrobium seedling is taken out from vial, clean the medium of root, soaking airing after 5 minutes in 0.1% liquor potassic permanganate, is cultivating container with little plastic basin, is cultivation matrix with the bog moss that soaked, keep suitably ventilating and enough humidity, the survival rate of transplanting all can reach 95%-100%.Adopt chamber planting during winter, temperature maintenance is more than 15 ℃, otherwise easily causes fallen leaves even dead.
The autumn dendrobium seedling that method provided by the invention is produced has the genetic stability height, and it is fast to emerge, and seedling is strong, advantages such as seedling quality better, resistance, well-grown.Method of the present invention can successfully be carried out the tissue culture and the breeding fast of the autumn dendrobium high quality seedling of improved seeds; owing to be to adopt vegetative propagation; implementing this method only need have simple Plant Tissue Breeding equipment to carry out; be to utilize biotechnologys such as the totipotency of plant tissue cell and Plant Tissue Breeding to carry out the large-scale production of plant high quality seedling; has less investment, the advantage that output is high.
Embodiment
Embodiment 1
1, draws materials: the butterfly stem of noble dendrobium.
2, the sterilization of explant: get the tender shoots that the butterfly stem of noble dendrobium is given birth to tool 5-6 joint then, after removing blade, be cut into two sections after rinsing well with running water, on superclean bench with 70% alcohol-pickled 1 minute, again with 0.1% mercuric chloride solution sterilization 10 minutes, behind the aseptic water washing 5 times, blot with aseptic blotting paper, the stem section and the stem apex of clip 2-3 centimetre of band joint are inoculated on protocorm and the inducing clumping bud medium.
3, inducing of the protocorm and the bud of growing thickly: inducing culture is on MS+6-benzyl purine 2-5 mg/litre+methyl 0.2-0.5 mg/litre+15% Sucus Cocois medium, under cultivation temperature (26 ± 2) ℃ condition, left and right sides stem apex began growth in 10 days, there is sprouting to form on 20 days left and right sides stem Duan Jie and growth, when the 6-benzyl purine is that the stylish bastem of 4-5 portion has protocorm to produce.Medium contains sucrose 30 grams per liters, pH 5.5-5.8, agar 0.7%.Illuminance 1500～2000lx, illumination 12 hours/day.
4, adventitious buds proliferation: the bud of growing thickly that will utilize stem section and Shoot Tip Culture to obtain, cut off partial blade and transfer to the adventitious buds proliferation medium: MS+6-benzyl purine 1-5 mg/litre+methyl 0.2-0.5 mg/litre+15% Sucus Cocois, can induce protocorm and grow thickly bud formation.The bud of when 6-benzyl purine concentration is the 1-2 mg/litre, only growing thickly, no protocorm, the existing bud of growing thickly when 6-benzyl purine concentration is the 3-4 mg/litre, protocorm is arranged again, when 6-benzyl purine concentration is 5 mg/litre when above, the protocorm of generation is many, and the bud quantity of growing thickly that induces is few.Protocorm can be bred on higher 6-benzyl purine concentration medium, and protocorm can differentiate the bud of growing thickly when changing the medium that hangs down 6-benzyl purine concentration over to, and the bud of growing thickly can be bred on the medium of low 6-benzyl purine concentration.
5, strong plantlets and rootage is cultivated: adventitious buds proliferation is after many generations, the bud of growing thickly can be cut into single bud, be seeded in and carry out strong plantlets and rootage on the strong plantlets and rootage medium of 1/2MS medium+methyl 0.5-1.0 mg/litre+bananas juice 100 grams per liters+activated carbon 0.5 grams per liter and cultivate.Can form plant height 3-4 centimetre whole plant about 40 days.
6, test-tube seedling transplanting: be the favorable season of transplanting spring.During test-tube seedling transplanting, natural lighting lower refining seedling 5-10 days, butterfly stem of noble dendrobium seedling is taken out from vial, cleaning the medium of root, soak airing after 5 minutes in 0.1% liquor potassic permanganate, is cultivating container with little plastic basin, with the bog moss that soaked is cultivation matrix, temperature maintenance keeps suitably ventilating and enough humidity more than 15 ℃, and the survival rate of transplanting all can reach 90%.
Claims (4)
1, a kind of high quality germchit tissure culturing and rapid breeding method of Dendrobium sp with autumn dendrobium elite germplasm, utilizes sprouting to be explant, adopts unique medium to carry out adventitious buds proliferation and obtain high quality seedling, it is characterized in that this method may further comprise the steps:
1). material: autumn dendrobium elite germplasm;
2). the sterilization of explant: get the tender shoots that the autumn dendrobium is given birth to tool 5-6 joint then, after removing blade, after rinsing well, water is cut into two sections, on superclean bench with 70% alcohol-pickled 1 minute, again with 0.1% mercuric chloride solution sterilization 10 minutes, behind the aseptic water washing 5 times, blot with aseptic blotting paper, the stem section and the stem apex of clip 2-3 centimetre of band joint are inoculated on protocorm and the inducing clumping bud medium;
3). inducing of the protocorm and the bud of growing thickly: inducing culture is on MS+6-benzyl purine 1-5 mg/litre+methyl 0.2-0.5 mg/litre+15% Sucus Cocois medium, under 26 ± 2 ℃ of conditions of cultivation temperature, stem apex began growth in 10 days, there is sprouting to form on 20 days left and right sides stem Duan Jie and growth, when the 6-benzyl purine is that the stylish bastem of 4-5 portion has protocorm to produce, medium contains sucrose 30 grams per liters, pH5.5-5.8, agar 0.7%;
4). adventitious buds proliferation: the bud of growing thickly that will utilize stem section and Shoot Tip Culture to obtain, cut off partial blade and transfer to the adventitious buds proliferation medium: MS+6-benzyl purine 1-5 mg/litre+methyl 0.2-0.5 mg/litre+15% Sucus Cocois, can induce protocorm and grow thickly bud formation, protocorm can be bred on higher 6-benzyl purine concentration medium, protocorm can differentiate the bud of growing thickly when changing the medium that hangs down 6-benzyl purine concentration over to, and the bud of growing thickly can be bred on the medium of low 6-benzyl purine concentration;
5). strong plantlets and rootage is cultivated: adventitious buds proliferation is after many generations, the bud of growing thickly can be cut into single bud, be seeded in and carry out strong plantlets and rootage on the strong plantlets and rootage medium of 1/2 MS medium+methyl 0.5-1.0 mg/litre+bananas juice 100 grams per liters+activated carbon 0.5 grams per liter and cultivate, can form plant height 3-4 centimetre whole plant in 40 days;
6). test-tube seedling transplanting: during test-tube seedling transplanting, natural lighting lower refining seedling 5-10 days, autumn dendrobium seedling is taken out from vial, cleaning the medium of root, soak airing after 5 minutes in 0.1% liquor potassic permanganate, is cultivating container with little plastic basin, with the bog moss that soaked is cultivation matrix, keep suitably ventilating and enough humidity, adopt chamber planting during winter, temperature maintenance is more than 15 ℃.
2, according to the dendrobium high quality seedling method for quickly breeding of autumn described in the claim 1, the favorable season that it is characterized in that test-tube seedling transplanting in the step 6) is spring.
3,, it is characterized in that the condition of culture in the step is illuminance 1500～2000lx, illumination 12 hours/day according to the dendrobium high quality seedling method for quickly breeding of autumn described in the claim 1.
4, according to the dendrobium high quality seedling method for quickly breeding of autumn described in the claim 1, it is characterized in that described material is that the butterfly stem of noble dendrobium, the goat's horn stem of noble dendrobium, panda No. 1, panda No. 2, Thailand are white, Zhuan Fu, purple butterfly, paint face, material is all taken from the South China Botanical Garden.
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Cited By (13)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN100401874C (en) * | 2006-03-20 | 2008-07-16 | 中国热带农业科学院热带生物技术研究所 | Stereo trough type nutrition providing cultivating method for dendrobium |
| CN100586275C (en) * | 2008-01-18 | 2010-02-03 | 中国科学院昆明植物研究所 | Fast replication method for dendrobium |
| CN101213941B (en) * | 2008-01-18 | 2011-01-12 | 中国科学院昆明植物研究所 | Fast replication and in-vitro conservation method for dendrobium |
| CN101695272B (en) * | 2009-10-26 | 2011-03-30 | 重庆市中药研究院 | Natural reproduction method for dendrobium |
| CN101336605B (en) * | 2007-07-07 | 2011-07-20 | 赤水市信天中药产业开发有限公司 | Dendrobium nobi tissue culturing and planting method |
| CN101632392B (en) * | 2009-08-26 | 2012-05-23 | 温州医学院附属第二医院 | Application of bryophyte as disinfectant for plant explants, disinfectant composition as well as preparation method and use method thereof |
| CN102657097A (en) * | 2012-06-02 | 2012-09-12 | 上海市闵行区农业科学研究所 | In-vitro culture method for tender stem segments of dendrobium nobile |
| CN103385174A (en) * | 2013-07-17 | 2013-11-13 | 宁波神乙草生物科技有限公司 | Tissue culture and inoculation method for dendrobium officinale |
| CN105028206A (en) * | 2015-08-21 | 2015-11-11 | 广西壮族自治区药用植物园 | Tissue culture propagation method for Hainan dendrobe |
| CN105028207A (en) * | 2015-08-21 | 2015-11-11 | 广西壮族自治区药用植物园 | Tissue culture propagation method for dendrobium gibsonii lindl |
| CN105028192A (en) * | 2015-06-17 | 2015-11-11 | 临沧市云瑞堂生物科技有限公司 | Culture medium series for rapidly breeding dendrobium nobile seedlings and tissue culture method |
| CN105284623A (en) * | 2015-11-13 | 2016-02-03 | 广西壮族自治区药用植物园 | Cymbidium bicolor Lindl. tissue culturing breeding method |
| CN107047305A (en) * | 2017-04-26 | 2017-08-18 | 福建省农业科学院作物研究所 | A kind of quick breeding method for tissue culture of dendrobium seedling |
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| CN101461328B (en) * | 2009-01-08 | 2011-03-23 | 中国科学院华南植物园 | High-efficient propagation method of seedlings of Dendrobium officinale |
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2004
- 2004-12-29 CN CN 200410077729 patent/CN1284448C/en not_active Expired - Fee Related
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| Publication number | Priority date | Publication date | Assignee | Title |
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| CN100401874C (en) * | 2006-03-20 | 2008-07-16 | 中国热带农业科学院热带生物技术研究所 | Stereo trough type nutrition providing cultivating method for dendrobium |
| CN101336605B (en) * | 2007-07-07 | 2011-07-20 | 赤水市信天中药产业开发有限公司 | Dendrobium nobi tissue culturing and planting method |
| CN100586275C (en) * | 2008-01-18 | 2010-02-03 | 中国科学院昆明植物研究所 | Fast replication method for dendrobium |
| CN101213941B (en) * | 2008-01-18 | 2011-01-12 | 中国科学院昆明植物研究所 | Fast replication and in-vitro conservation method for dendrobium |
| CN101632392B (en) * | 2009-08-26 | 2012-05-23 | 温州医学院附属第二医院 | Application of bryophyte as disinfectant for plant explants, disinfectant composition as well as preparation method and use method thereof |
| CN101695272B (en) * | 2009-10-26 | 2011-03-30 | 重庆市中药研究院 | Natural reproduction method for dendrobium |
| CN102657097A (en) * | 2012-06-02 | 2012-09-12 | 上海市闵行区农业科学研究所 | In-vitro culture method for tender stem segments of dendrobium nobile |
| CN103385174A (en) * | 2013-07-17 | 2013-11-13 | 宁波神乙草生物科技有限公司 | Tissue culture and inoculation method for dendrobium officinale |
| CN103385174B (en) * | 2013-07-17 | 2015-11-18 | 宁波神乙草生物科技有限公司 | Candidum tissue culturing inoculation method |
| CN105028192A (en) * | 2015-06-17 | 2015-11-11 | 临沧市云瑞堂生物科技有限公司 | Culture medium series for rapidly breeding dendrobium nobile seedlings and tissue culture method |
| CN105028206A (en) * | 2015-08-21 | 2015-11-11 | 广西壮族自治区药用植物园 | Tissue culture propagation method for Hainan dendrobe |
| CN105028207A (en) * | 2015-08-21 | 2015-11-11 | 广西壮族自治区药用植物园 | Tissue culture propagation method for dendrobium gibsonii lindl |
| CN105284623A (en) * | 2015-11-13 | 2016-02-03 | 广西壮族自治区药用植物园 | Cymbidium bicolor Lindl. tissue culturing breeding method |
| CN107047305A (en) * | 2017-04-26 | 2017-08-18 | 福建省农业科学院作物研究所 | A kind of quick breeding method for tissue culture of dendrobium seedling |
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| CN1284448C (en) | 2006-11-15 |
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