CN1535571A - Sterile sowing and tissue culture method of tree orchid - Google Patents
Sterile sowing and tissue culture method of tree orchid Download PDFInfo
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- CN1535571A CN1535571A CNA031141781A CN03114178A CN1535571A CN 1535571 A CN1535571 A CN 1535571A CN A031141781 A CNA031141781 A CN A031141781A CN 03114178 A CN03114178 A CN 03114178A CN 1535571 A CN1535571 A CN 1535571A
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Abstract
The present invention relates to a sterile sowing and tissue culture method of epidendrum radians. The invention adopts artificial nutrient liquor and utilizes sterile sowing method to oblain lots of test-tube seedlings, and utilizes the stem apex or stipes of sterile seedling-plant to make multiplication of clustered bud, then utilizes rootage and culture to form whole plant.
Description
Technical field
The present invention relates to aseptic seeding and the method for tissue culture of tree orchid.
Technical background
Tree orchid belongs to (Epidendrum) and has another name called post lobe Cymbidium, is one of genus maximum in the orchid family, and this platymiscium whole world has kind more than 1000 approximately, grows nonparasitically upon another plant or give birth on ground, and fecund can be cultivated in high temperature, middle temperature or low temperature greenhouse according to the different habits of kind in tropical America.Some kind has resting stage, and some does not have tangible resting stage.The variation of its stem is also bigger, the tool pseudobulb that has, and the reedy upright stalk of the tool that has, there is the minority leaf on the top of pseudobulb.The scape of tree orchid platymiscium is born in the stem top, and flower has several to many, spends large-scalely or small-sized, and sepal and petal are all very carefully narrow, but the lip hypertrophy, base portion is recessed deeply, and the stamen post is very flourishing, so a post lobe Cymbidium.Tree orchid platymiscium in a great variety, wherein numerous species has very high ornamental value.Tree orchid (Epidendrum radians) is more representative kind.
Tree orchid (Epidendrum radians) is an epiphytic orchid, originate in Mexico, stem elongated cylindrical and multi-branched, on it give birth to most linen aerial roots, blade is elongated, the flower cerise, tens flowers are lined up the top that raceme is born in a long scape, the florescence spring and summer is to late fall, and the florescence is long, has very high ornamental value.The breeding of tree orchid can be adopted cuttage and plant division, but reproduction speed is slow, and the tree orchid seed is incomplete owing to embryonic development, extremely difficult sprouting the under the nature.
At present, the aseptic seeding of tree orchid and the report of tissue culture and large-scale production and patent application are not arranged both at home and abroad.
Summary of the invention
It is incomplete because of embryonic development to the objective of the invention is to solve the tree orchid seed, the problem that is difficult to sprout under nature, but the method for a kind of large-scale breeding tree orchid seedling of exploitation.
The present invention adopts artificial nutrient liquid to utilize the method for aseptic seeding to obtain a large amount of test-tube plantlets, and utilizes the grow thickly propagation of bud and form whole plant by culture of rootage of the stem apex of aseptic seedling or stipes, has realized purpose of the present invention.
Technical characterictic of the present invention is, comprises the following steps:
(1) material is selected: the maternal plant of choosing robust growth carries out artificial pollination when blooming, and the fruit after 110-130 days of pollinating is as explant;
(2) aseptic seeding: be placed among the liquor natrii hypochloritis of 8%-15% sterilization second with the alcohol-pickled 20-40 of 70%-80% 10-30 minute, fruit is cut in 4-5 back of aseptic water washing, with transfer needle the uniform broadcasting of white powder embryo is inoculated in the medium, 100 milliliters of the Sucus Cocois that contains in every liter of the described medium, agar 7 grams, sucrose 20 grams, all the other compositions are N
6
(3) tissue culture: utilize the stem apex or the stipes of the aseptic seedling of above-mentioned steps growth in medium, to form the bud of growing thickly, contain 0.5 milligram of 6-benzyl purine in every liter of the described medium, agar 7 grams, sucrose 20 grams, all the other compositions are MS, contain 0.5 milligram of 6-benzyl purine, agar 7 grams in the subculture propagation in medium, every liter of described medium, sucrose 30 grams, all the other compositions are MS, and the bud of growing thickly forms whole plant with medium culture when taking root, and contain 0.2 milligram of methyl in every liter of the described medium, bananas juice 100 grams, agar 7 grams, sucrose 20 grams, remaining composition is 1/2MS;
(4) test-tube seedling transplanting: when test-tube plantlet can grow to 3-4 centimetre high, transferred to the natural daylight lower refining seedling 7-10 days, then it is taken out from vial, clean the medium of root, move into volume ratio and be in the mixed-matrix of 1: 1 peat and thick coconut husk.
At above-mentioned the medium pH 5.4-5.6 described in steps, cultivation temperature (26 ± 2) ℃, illuminance 1500～2000lx, illumination 12 hours/day.
In above-mentioned medium in steps, add the curable medium of agar.
In the step of described aseptic seeding, described Sucus Cocois is that the Sucus Cocois that takes out from fresh coconut is through multilayer filtered through gauze gained.Described N
6Be international medium, its composition and compound method are referring to Tan Wencheng, Dai Cegang chief editor, " ornamental plants tissue culture technique ", Beijing: China Forest publishing house, 1991.
In the step of described aseptic seeding, a general fruit can be broadcast the 3-5 bottle, and 20-40 days left and right sides embryo germinations formed whole plant about 55-65 days; The too close and seedling of the aseptic bacterium of growing directly from seeds hour can be cultivated strong sprout on identical medium in medium.
In the step of described tissue culture, described bananas juice is to be smashed with juice extractor by banana peeling back to form.Described 1/2MS reduces by half macroelement among the MS and trace element, other components unchanged and the medium that forms, described MS is international medium, its composition and compound method are edited referring to Tan Wencheng, Dai Cegang, " ornamental plants tissue culture technique ", Beijing: China Forest publishing house, 1991.
In the step of described test-tube seedling transplanting, keep suitably ventilating and enough humidity, the survival rate of transplanting all can reach more than 90%.
The experiment condition and the culture medium prescription of the best of the present invention are seen embodiment.
The present invention has following advantage:
(1) increment multiple height: the increment of the bud of growing thickly can reach 3-5 doubly;
(2) the survival rate height of Yi Zaiing: survival rate can reach more than 90%;
(3) equipment is simple: only need simple Plant Tissue Breeding equipment to carry out;
(4) emerge soon, stability is high, and seedling is strong, seedling quality better, resistance, well-grown;
(5) can be used as tree orchid belong to other kind, belong in or with the method for other aseptic seeding of bigenering and tissue culture.
Embodiment
Embodiment 1: aseptic seeding of tree orchid and tissue culture propagating
1. the maternal plant of choosing robust growth carries out artificial pollination when blooming, and pollinates and gets fruit after 120 days as explant;
2. with 75% alcohol-pickled 30 seconds being placed among 10% the liquor natrii hypochloritis sterilization 20 minutes, cut fruit behind the aseptic water washing 4 times, with transfer needle the uniform broadcasting of white powder embryo is inoculated in the medium, 100 milliliters of the Sucus Cocois that contains in every liter of the described medium, agar 7 grams, sucrose 20 grams, all the other compositions are N
6, a carpostrote 4 bottles, 30 days embryo germinations formed whole plant in 60 days;
Illustrate: described Sucus Cocois is that the Sucus Cocois that takes out from fresh coconut is through multilayer filtered through gauze gained.Described medium pH 5.5, cultivation temperature (26 ± 2) ℃, illuminance 1500～2000lx, illumination 12 hours/day.
3. utilize the stem apex or the stipes of the aseptic seedling of above-mentioned steps growth in medium, to form the bud of growing thickly, contain 0.5 milligram of 6-benzyl purine in every liter of the described medium, agar 7 grams, sucrose 20 grams, all the other compositions are MS, subculture propagation is 5 times in medium, contains 0.5 milligram of 6-benzyl purine, agar 7 grams in every liter of the described medium, sucrose 30 grams, all the other compositions are MS, and the bud of growing thickly forms whole plant with medium culture when taking root, and contain 0.2 milligram of methyl in every liter of the described medium, bananas juice 100 grams, agar 7 grams, sucrose 20 grams, remaining composition is 1/2MS;
Illustrate: described bananas juice is smashed with juice extractor by banana peeling back and is formed.Described medium pH 5.5, cultivation temperature (26 ± 2) ℃, illuminance 1500～2000lx, illumination 12 hours/day.
4. test-tube seedling transplanting: when test-tube plantlet can grow to 3-4 centimetre high, transferred to the natural daylight lower refining seedling 10 days, then it is taken out from vial, clean the medium of root, move into volume ratio and be in the mixed-matrix of 1: 1 peat and thick coconut husk.
5. result: the survival rate of transplanting is 95%.
Claims (2)
1. aseptic seeding of a tree orchid and method for tissue culture is characterized in that comprising 4 steps:
(1) material is selected: the fruit after 110-130 days of pollinating is as explant;
(2) aseptic seeding: be placed among the liquor natrii hypochloritis of 8%-15% sterilization second with the alcohol-pickled 20-40 of 70%-80% 10-30 minute, fruit is cut in 4-5 back of aseptic water washing, with transfer needle the uniform broadcasting of white powder embryo is inoculated in the medium, 100 milliliters of the Sucus Cocois that contains in every liter of the described medium, agar 7 grams, sucrose 20 grams, all the other compositions are N
6
(3) tissue culture: utilize the stem apex or the stipes of the aseptic seedling of above-mentioned steps growth in medium, to form the bud of growing thickly, contain 0.5 milligram of 6-benzyl purine in every liter of the described medium, agar 7 grams, sucrose 20 grams, all the other compositions are MS, contain 0.5 milligram of 6-benzyl purine, agar 7 grams in the subculture propagation in medium, every liter of described medium, sucrose 30 grams, all the other compositions are MS, and the bud of growing thickly forms whole plant with medium culture when taking root, and contain 0.2 milligram of methyl in every liter of the described medium, bananas juice 100 grams, agar 7 grams, sucrose 20 grams, remaining composition is 1/2MS;
(4) test-tube seedling transplanting: when test-tube plantlet can grow to 3-4 centimetre high, transferred to the natural daylight lower refining seedling 7-10 days, then it is taken out from vial, clean the medium of root, move into volume ratio and be in the mixed-matrix of 1: 1 peat and thick coconut husk;
At above-mentioned the medium pH 5.4-5.6 described in steps, cultivation temperature (26 ± 2) ℃, illuminance 1500～2000lx, illumination 12 hours/day.
2. aseptic seeding of a kind of tree orchid according to claim 1 and method for tissue culture is characterized in that:
(1) during described material is selected, chooses the fruit of pollination after 120 days as explant;
(2) in the step of described aseptic seeding, with 75% alcohol-pickled 30 seconds being placed among 10% the liquor natrii hypochloritis sterilization 20 minutes;
(3) in the step of described test-tube seedling transplanting, when test-tube plantlet can grow to 3-4 centimetre high, transferred to the natural daylight lower refining seedling 10 days.
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| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN 03114178 CN1218629C (en) | 2003-04-10 | 2003-04-10 | Sterile sowing and tissue culture method of tree orchid |
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| Application Number | Priority Date | Filing Date | Title |
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| CN 03114178 CN1218629C (en) | 2003-04-10 | 2003-04-10 | Sterile sowing and tissue culture method of tree orchid |
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| CN1535571A true CN1535571A (en) | 2004-10-13 |
| CN1218629C CN1218629C (en) | 2005-09-14 |
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Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101066041B (en) * | 2007-06-08 | 2011-01-12 | 中国科学院昆明植物研究所 | Aseptic seeding and tissue culture process of cymbidium lianpan |
| CN102884979A (en) * | 2012-09-17 | 2013-01-23 | 贵州省林业科学研究院 | Quick tissue propagation method of eria coronaria |
| CN107466858A (en) * | 2017-09-25 | 2017-12-15 | 贵州大学 | A kind of bamboo shoot orchid species seedling rapid propagation method |
| CN115644058A (en) * | 2022-09-23 | 2023-01-31 | 广西壮族自治区农业科学院 | Method for promoting orchidaceae plant seeds to quickly grow seedlings |
-
2003
- 2003-04-10 CN CN 03114178 patent/CN1218629C/en not_active Expired - Fee Related
Cited By (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101066041B (en) * | 2007-06-08 | 2011-01-12 | 中国科学院昆明植物研究所 | Aseptic seeding and tissue culture process of cymbidium lianpan |
| CN102884979A (en) * | 2012-09-17 | 2013-01-23 | 贵州省林业科学研究院 | Quick tissue propagation method of eria coronaria |
| CN102884979B (en) * | 2012-09-17 | 2014-04-16 | 贵州省林业科学研究院 | Quick tissue propagation method of eria coronaria |
| CN107466858A (en) * | 2017-09-25 | 2017-12-15 | 贵州大学 | A kind of bamboo shoot orchid species seedling rapid propagation method |
| CN107466858B (en) * | 2017-09-25 | 2019-12-03 | 贵州大学 | A kind of bamboo shoot orchid species seedling rapid propagation method |
| CN115644058A (en) * | 2022-09-23 | 2023-01-31 | 广西壮族自治区农业科学院 | Method for promoting orchidaceae plant seeds to quickly grow seedlings |
| CN115644058B (en) * | 2022-09-23 | 2023-11-21 | 广西壮族自治区农业科学院 | Method for promoting quick seedling formation of orchid seeds |
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| Publication number | Publication date |
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| CN1218629C (en) | 2005-09-14 |
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