CN1284444C - Sterile seeding and tissue cultivating technology for Vanda - Google Patents
Sterile seeding and tissue cultivating technology for Vanda Download PDFInfo
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- CN1284444C CN1284444C CN 200410050905 CN200410050905A CN1284444C CN 1284444 C CN1284444 C CN 1284444C CN 200410050905 CN200410050905 CN 200410050905 CN 200410050905 A CN200410050905 A CN 200410050905A CN 1284444 C CN1284444 C CN 1284444C
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Abstract
The present invention relates to a sterile seeding and tissue cultivating technology for Vanda. Vanda, Cattleya hybrida, Dendrobium and Phalaenopsis amabilis are together known as four main kinds of ornamental orchids. The conventional propagating method for Vanda has a tillering method, a cuttage method, etc., but the propagation speed is low. The seeds of Vanda are extremely difficult to germinate in a natural state because the growth of embryos is incomplete, and therefore, as for cross breeding and the propagation of initial species, the sterile seeding is one of the most effective propagating methods. In the present invention, an artificial nutrition medium is adopted, a large amount of test-tube seedlings can be obtained by the sterile seeding method, and the stem tips or the stem nodes of sterile seedlings can be used for proliferating multiple shoots and can form complete plants through rooting cultivation. The present invention provides a method for sterile seeding and tissue cultivating propagation of Vanda, and the culture medium used, and has the advantages of strong operability and high application value.
Description
Technical field
The present invention relates to the aseptic seeding and the method for tissue culture of vanda.
Background technology
Vanda (Vanda) has another name called all ages orchid, is called as " four view and admire orchid greatly " with Bowring cattleya, dendrobium, Moth orchid.Vanda originates in the torrid zone, Asia and subtropical zone, and there is more than 70 initial species in this platymiscium whole world, more than 600 crossbreed, and China has found 14 kinds and 1 mutation, mainly is distributed in southern areas.
Vanda is the cattleya of single shaft class stem, and tangible stem and aerial root are arranged.Its root is sturdy, not only can dependence itself flourishing root system make whole strain plant grow nonparasitically upon another plant on other plant or rock and support the upwards growth of its stem, and flourishing root system also is responsible for the absorption of moisture and nutriment, it is green that particularly some kind butt is exposed to outside one-tenth, contain chlorophyll, can equally with blade carry out photosynthesis; Some kind under the condition of high humility, eustipes part can give birth to hundreds of white root, just like Beauty Beard one pure white beard, and for example multifarious ribbon has a distinctive style, and is very noticeable.The blade of vanda is lined up two row in the neat close life in the both sides of stem, and like kaffir lily, but leaf is longer, band shape.The flower of vanda is a raceme, usually extract out from axil, each axil of vanda that south east asia such as Thailand, Singapore is produced nearly all can be extracted a flower out, every strain can be opened 15 ~ 20 sometimes, and spending of every flower of vanda is big and intensive, generally spends 10 ~ 20, every flower can open for 2 ~ 3 weeks, usually open successively on from descending, so, when temperature is low, can reach two wheat harvesting periods even every flower all can be opened a wheat harvesting period under hot conditions.
Although the structure of vanda flower is similar substantially to other orchid, the ratio of each several part differs bigger.As lip and the petal of orchid very flourishing and remarkable, but the crossbreed petal of vanda is less, lip is littler, on viewing and admiring, do not accounted for critical role, 3 pieces of sepals are then flourishing especially, especially two pieces of side sepals are bigger, and have multiple shapes such as ellipse, circle, microscler, triangle, become the emphasis of appreciation.The vanda calyx is bright in luster, colorful, and rare dark brown of pink, yellow, aubergine, pure white and other orchid and sky blue etc. are arranged, and the spot that has shades of colour and shape on the flower is very gorgeous, the ornamental value height.Can be used for potted plant or hang basin and view and admire, also be widely used in cut-flower, on China market, also may become the blue focus of selling in a new torrid zone after Moth orchid, the hybrid cymbidium.
At present, the vanda of selling on the flowers market mostly is crossbreed, but part initial species such as Da Hua vanda (Vandacoerulea) wait and itself also have very high ornamental value.Particularly importantly the hybridization affinity of vanda is strong, can belong to hybridizing with more than 20 of the orchid families obtaining the high new varieties of ornamental value.
The conventional propagation method of vanda has tillers and cuttage etc., but reproduction speed is slow, and the vanda seed is because embryonic development is incomplete, extremely difficult sprouting the under the nature.Therefore, no matter be the crossbreeding or the breeding of initial species, adopting aseptic seeding is one of the most effective propagation method.
The present invention adopts the artificial alimentation base to utilize the method for aseptic seeding can obtain a large amount of test-tube plantlets, and utilizes can the grow thickly propagation of bud and can form whole plant by culture of rootage of the stem apex of aseptic seedling or stipes.
At present, the aseptic seeding of vanda and the report of tissue culture and large-scale production and patent application are not arranged both at home and abroad.
The invention provides the aseptic seeding of vanda and the method and the employed medium of tissue culture propagating, workable, the using value height.
Summary of the invention
The method that the objective of the invention is to adopt the artificial alimentation base to utilize aseptic seeding can obtain a large amount of test-tube plantlets, and utilizes can the grow thickly propagation of bud and can form whole plant by culture of rootage of the stem apex of aseptic seedling or stipes.The invention provides the aseptic seeding of vanda and the method and the employed medium of tissue culture propagating, workable, the using value height.
The inventive method characteristics are:
The breeding of vanda can be adopted cuttage and plant division, but reproduction speed is slow, and the vanda seed is incomplete owing to embryonic development, extremely difficult sprouting the under the nature.The present invention adopts the artificial alimentation base to utilize the method for aseptic seeding can obtain a large amount of test-tube plantlets, and utilizes can the grow thickly propagation of bud and can form whole plant by culture of rootage of the stem apex of aseptic seedling or stipes.
The method of the invention can successfully be carried out the aseptic seeding and the tissue-culturing quick-propagation of vanda seedling.Has less investment, the characteristics that output is high.
Implementing present technique only need have simple Plant Tissue Breeding equipment to carry out.
The aseptic seeding and the method for tissue culture of vanda of the present invention comprise the steps:
1. material: adopting Da Hua vanda (Vanda coerulea) and bird tongue orchid (Ascocentrum ampullaceum) is the parent, the maternal plant of choosing robust growth when blooming carries out the pollination self of Da Hua vanda and carries out the Da Hua vanda and the blue hybridization of bird tongue, and pollination back fruit 120-150 days is basic to be used for sowing as explant when ripe.
2. aseptic seeding: get the fruit that is mature on the whole, with 75% alcohol-pickled 30 seconds being placed in 0.1% the mercuric chloride solution sterilization 20 minutes, cut fruit behind the aseptic water washing 4～5 times, the white powder seed is inoculated into seed germination medium: V﹠amp with transfer needle; W medium+6-benzyl purine 0.2-1 mg/litre+methyl 0.2-1 mg/litre+10-20% coconut milk.30-40 days left and right sides embryo germinations formed whole plant in 60 days, and the too close and seedling of aseptic seedling hour can carry out strong seedling culture on identical medium in medium.
3. adventitious buds proliferation: for obtaining more seedling, utilize the stem apex or the stipes cultivation of aseptic seedling can form the bud of growing thickly on the proliferated culture medium of the coconut milk of MS medium+6-benzyl purine 1.0-2.0 mg/litre+methyl 0.5-1.0 mg/litre+10-20%, the bud of growing thickly can carry out shoot proliferation on identical medium.
4. strong plantlets and rootage: will sow the bud of growing thickly that the plant that forms or shoot proliferation obtain and cut into single bud, and be seeded in to contain on the root media of spending precious (Hyponex 1) 3 grams per liters+sucrose 15 grams per liters+bananas juice 100 grams per liters+potato 100 a grams per liters+6-benzyl purine 0.1-0.5 mg/litre+methyl 0.5-2 mg/litre+activated carbon 0.5-2 grams per liter and carry out culture of rootage.Can form plant height 3-4 centimetre whole plant about 40 days.
5. test-tube seedling transplanting: when test-tube plantlet can grow to 3-4 centimetre high, transferred to the natural daylight lower refining seedling 7-14 days, then it is taken out from vial, clean the medium of root, move in the mixed-matrix of peat and thick coconut husk, keep suitably ventilating enough humidity, the survival rate of transplanting all can reach more than 95%.
Used medium additional saccharose 20 grams per liters all in experiment, pH5.4-5.6, agar 0.7%.Cultivation temperature (26 ± 2) ℃, illuminance 1500～2000lx, illumination 12 hours/day.
The method of the invention compared with prior art has emerges soon, and stability is high, and seedling is strong, advantages such as seedling quality better, resistance, well-grown.The present invention simultaneously can be as other kind of Vanda, belong in or with the method for other aseptic seeding of bigenering and tissue culture.
Embodiment:
Embodiment 1:
1. material: Da Hua vanda (Vanda coerulea) maternal plant of choosing robust growth carries out pollination self when blooming, and pollination back fruit is in the time of 140 days, and fruit is mature on the whole, and is used for sowing with the seed in the ripening fruits as explant.
2. aseptic seeding: get the fruit that is mature on the whole, with 75% alcohol-pickled 30 seconds being placed in 0.1% the mercuric chloride solution sterilization 20 minutes, cut fruit behind the aseptic water washing 4 times, the white powder seed is inoculated into seed germination medium: V﹠amp with transfer needle; W medium+6-benzyl purine 1.0 mg/litre+methyl 0.5 mg/litre+10%-20% coconut milk.40 days left and right sides embryo germinations formed whole plant about 60 days, and the too close and seedling of aseptic seedling hour can carry out strong seedling culture on identical medium in medium.
3. adventitious buds proliferation: for obtaining more seedling, utilize the stem apex or the stipes cultivation of aseptic seedling can form the bud of growing thickly on the proliferated culture medium of the coconut milk of MS medium+6-benzyl purine 2.0 mg/litre+methyl 1.0 mg/litre+20%, the bud of growing thickly can carry out shoot proliferation on identical medium.
4. strong plantlets and rootage: will sow the bud of growing thickly that the plant that forms or shoot proliferation obtain and cut into single bud, and be seeded in to contain on the root media of spending precious (Hyponex 1) 3 grams per liters+sucrose 15 grams per liters+bananas juice 100 grams per liters+potato 100 grams per liters+6-benzyl purine 0.5 mg/litre+methyl 2 a mg/litre+activated carbon 1.0. grams per liter and carry out culture of rootage.Can form plant height 3-4 centimetre whole plant about 40 days.
5. test-tube seedling transplanting: when test-tube plantlet can grow to 3-4 centimetre high, transferred to the natural daylight lower refining seedling 7 days, then it is taken out from vial, clean the medium of root, move in the mixed-matrix of peat and thick coconut husk, keep suitably ventilating and enough humidity, the survival rate of transplanting all can reach more than 95%.
Used medium additional saccharose 20 grams per liters all in experiment, pH5.4-5.6, agar 0.7%.Cultivation temperature (26 ± 2) ℃, illuminance 1500～2000lx, illumination 12 hours/day.
Embodiment 2:
1. material: adopting Da Hua vanda (Vanda coerulea) and bird tongue orchid (Ascocentrum ampullaceum) be the parent, and the maternal plant of choosing robust growth when blooming hybridizes, and is used for sowing as explant during the basic maturation of pollination back fruit 130 days.
2. aseptic seeding: get the fruit that is mature on the whole, with 75% alcohol-pickled 30 seconds being placed in 0.1% the mercuric chloride solution sterilization 20 minutes, cut fruit behind the aseptic water washing 5 times, the white powder seed is inoculated into seed germination medium: V﹠amp with transfer needle; W medium+6-benzyl purine 1.0 mg/litre+methyl 0.2 mg/litre+10% coconut milk.30 days left and right sides embryo germinations formed whole plant about 60 days, and the too close and seedling of aseptic seedling hour can carry out strong seedling culture on identical medium in medium.
3. adventitious buds proliferation: for obtaining more seedling, utilize the stem apex or the stipes cultivation of aseptic seedling can form the bud of growing thickly on the proliferated culture medium of the coconut milk of MS medium+6-benzyl purine 1.0 mg/litre+methyl 0.5 mg/litre+10%, the bud of growing thickly can carry out shoot proliferation on identical medium.
4. strong plantlets and rootage: will sow the bud of growing thickly that the plant that forms or shoot proliferation obtain and cut into single bud, and be seeded in to contain on the root media of spending precious (Hyponex 1) 3 grams per liters+sucrose 15 grams per liters+bananas juice 100 grams per liters+potato 100 grams per liters+6-benzyl purine 0.2 mg/litre+methyl 1.0 mg/litre+activated carbon 1.5 grams per liters and carry out culture of rootage.Can form plant height 3-4 centimetre whole plant about 40 days.
5. test-tube seedling transplanting: when test-tube plantlet can grow to 3-4 centimetre high, transferred to the natural daylight lower refining seedling 7 days, then it is taken out from vial, clean the medium of root, move in the mixed-matrix of peat and thick coconut husk, keep suitably ventilating and enough humidity, the survival rate of transplanting all can reach 100%.
Used medium additional saccharose 20 grams per liters all in experiment, pH5.4-5.6, agar 0.7%.Cultivation temperature (26 ± 2) ℃, illuminance 1500～2000lx, illumination 12 hours/day.
Claims (1)
1, a kind of aseptic seeding of vanda and method for tissue culture is characterized in that this method may further comprise the steps:
1), material: adopting Da Hua vanda and bird tongue orchid is the parent, the maternal plant of choosing robust growth when blooming carries out the pollination self of Da Hua vanda and carries out the Da Hua vanda and the blue hybridization of bird tongue, during 120-150 days maturations of pollination back fruit, be used for sowing as explant;
2), aseptic seeding: get the fruit that is mature on the whole, with 75% alcohol-pickled 30 seconds being placed in 0.1% the mercuric chloride solution sterilization 20 minutes, cut fruit behind the aseptic water washing 4～5 times, with transfer needle the white powder seed is inoculated into 30-40 days embryo germinations in the seed germination medium, formed whole plant in 60 days, the too close and seedling of aseptic seedling hour carries out strong seedling culture on identical medium in medium; 26 ± 2 ℃ of cultivation temperature, illuminance 1500～2000lx, illumination 12 hours/day, the seed germination medium is: V﹠amp; W medium+6-benzyl purine 0.2-1 mg/litre+methyl 0.2-1 mg/litre+10-20% coconut milk, additional saccharose 20 grams per liters, pH5.4-5.6, agar 0.7%;
3), adventitious buds proliferation: for obtaining more seedling, utilize the stem apex or the stipes cultivation of aseptic seedling on proliferated culture medium, to form the bud of growing thickly, the bud of growing thickly carries out shoot proliferation on identical medium, 26 ± 2 ℃ of cultivation temperature, illuminance 1500～2000lx, illumination 12 hours/day, proliferated culture medium is: the coconut milk of MS medium+6-benzyl purine 1.0-2.0 mg/litre+methyl 0.5-1.0 mg/litre+10-20%, additional saccharose 20 grams per liters, pH 5.4-5.6, agar 0.7%;
4), strong plantlets and rootage: will sow the bud of growing thickly that the plant that forms or shoot proliferation obtain and cut into single bud, and be seeded in and carry out culture of rootage on the root media; Formed the whole plant of 3-4 centimetre of plant height in 40 days; 26 ± 2 ℃ of cultivation temperature, illuminance 1500～2000lx, illumination 12 hours/day, root media is: contain and spend precious 3 grams per liters+sucrose 15 grams per liters+bananas juice 100 grams per liters+potato 100 grams per liters+6-benzyl purine 0.1-0.5 mg/litre+methyl 0.5-2 mg/litre+activated carbon 0.5-2 grams per liter, additional saccharose 20 grams per liters, pH 5.4-5.6, agar 0.7%;
5), test-tube seedling transplanting: when test-tube plantlet grows to 3-4 centimetre high, transferred to the natural daylight lower refining seedling 7-14 days, then it is taken out from vial, clean the medium of root, move in the mixed-matrix of peat and thick coconut husk, keep ventilating and enough humidity.
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| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN 200410050905 CN1284444C (en) | 2004-07-30 | 2004-07-30 | Sterile seeding and tissue cultivating technology for Vanda |
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| Application Number | Priority Date | Filing Date | Title |
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| CN 200410050905 CN1284444C (en) | 2004-07-30 | 2004-07-30 | Sterile seeding and tissue cultivating technology for Vanda |
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| CN1586164A CN1586164A (en) | 2005-03-02 |
| CN1284444C true CN1284444C (en) | 2006-11-15 |
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Families Citing this family (9)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1320853C (en) * | 2005-04-22 | 2007-06-13 | 云南农业大学 | Method for breeding precious cross bred orchid |
| CN101180950B (en) * | 2007-12-26 | 2010-12-08 | 浙江森禾种业股份有限公司 | Tissue cultivation rapid breeding method of spring dendrobium stem |
| CN103004604B (en) * | 2012-12-27 | 2013-12-25 | 福建农林大学 | Breeding method for vanda |
| CN104054569A (en) * | 2014-05-30 | 2014-09-24 | 霍山宝信园石斛开发有限公司 | Selective breeding method production process of wild dendrobium officinale |
| CN104054574A (en) * | 2014-05-30 | 2014-09-24 | 霍山宝信园石斛开发有限公司 | Selective breeding method production process of dendrobium moniliforme |
| CN105210871A (en) * | 2015-10-12 | 2016-01-06 | 广西壮族自治区药用植物园 | A kind of pure color all ages blue tissue culture method for breeding |
| CN106613992A (en) * | 2017-01-10 | 2017-05-10 | 中国林业科学研究院林业研究所 | Medium for aseptic sowing and rapid seedling of single-flower Paphiopedilum |
| CN106718933A (en) * | 2017-01-23 | 2017-05-31 | 中国林业科学研究院林业研究所 | Spend more the culture medium of class pocket orchid aseptic seeding and sprouting and rooting |
| CN107646591A (en) * | 2017-09-30 | 2018-02-02 | 江苏农林职业技术学院 | A kind of bletilla striata direct sowing and seedling matrix and the method cultivated using the matrix |
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