CN101066041B - Aseptic seeding and tissue culture process of cymbidium lianpan - Google Patents
Aseptic seeding and tissue culture process of cymbidium lianpan Download PDFInfo
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- CN101066041B CN101066041B CN2007100659413A CN200710065941A CN101066041B CN 101066041 B CN101066041 B CN 101066041B CN 2007100659413 A CN2007100659413 A CN 2007100659413A CN 200710065941 A CN200710065941 A CN 200710065941A CN 101066041 B CN101066041 B CN 101066041B
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- Breeding Of Plants And Reproduction By Means Of Culturing (AREA)
Abstract
The aseptic seeding and tissue culture process of Cymbidium lianpan includes seed germination of Cymbidium lianpan material, proliferating culturing, test tube plantlet inducing, hardening off, transplanting and other steps. The present invention provides fast propagation way for Cymbidium lianpan as rare orchid variety with very high ornamental value and economic value.
Description
Technical field:
The invention belongs to plant biotechnology field, particularly, relate to aseptic seeding and the method for tissue culture of a kind of orchid " lotus lobe orchid ".
Background technology:
The blue special product of lotus lobe is rare orchid in western part, Yunnan.Its form grace, plant type are beautiful; The blade four seasons are green for a long time; Spend luicd and elegant graceful delicate fragrance, pattern is abundant, has very high ornamental value and economic worth.It is the orchid treasure that many kinds are wherein arranged, and as element etc. in slight snow, a thousand pieces of gold difficulty is asked.The blue seed of lotus lobe is little, extremely difficult the germination, and division propagation is slow again.So set up the tissue cultivating seedling clone, be valuable and realistic meaning for the breeding of the blue treasure of lotus lobe provides an approach faster.So far do not see the seed asepsis sprouting of lotus lobe orchid and the report of tissue culture in the prior art as yet.
Summary of the invention:
The object of the present invention is to provide aseptic seeding and the method for tissue culture of a kind of orchid " lotus lobe orchid ".
In order to realize above-mentioned purpose of the present invention, the invention provides following technical scheme:
The aseptic seeding and the method for tissue culture of " lotus lobe orchid " are got lotus lobe orchid, and process seed germination, enrichment culture, test-tube plantlet are induced, the acclimatization and transplants step, and the blue seed of lotus lobe is broadcast at minimal medium+BA 5mgL
-1+ NAA 5mgL
-1Seed culture medium on cultivated 6-7 month, differentiate root-like stock, root-like stock is inserted minimal medium+BA3mgL
-1+ NAA5mgL
-1Proliferated culture medium on carry out enrichment culture, treat that root-like stock changes green gradually, the part bud differentiates, the agglomerate that will have bud and root-like stock clump is cut into 1cm
2Size inserts minimal medium+BA3mgL
-1+ IAA0.2mgL
-1The bud inducing culture on, root-like stock differentiates clump bud, and the clump sorite is cut into the 2-3 sprout tuber, is seeded in minimal medium+NAA1mgL
-1+ banana puree 20%, on the root media of pH5.5 3 months, treat that bud grows several roots of 1-, seedling grows to 5cm when above, is placed on 40 μ molm
-1S
-1Light intensity lower refining seedling 15-30 days, when vane extension, plant grow tall and are healthy and strong, seedling is taken out, transplanting is in moistening sterile sphagna, the lid plastic foil is preserved moisture, keep about 25 ℃ of environmental temperature, ventilation is preserved moisture, treat that test-tube plantlet grows young leaves or new root can be removed coverlay, Routine Management, the used minimal medium of above-mentioned steps are and spend precious No. 1 is N: P: K=7: product Hypenox1 2.5g+MS organic principle, molysite and the trace element+AC 0.5gL of 6: 19 U.S. Hyponex chemical company
-1+ S 2%, and agar solidifies; Cultivation temperature (26 ± 2) ℃, continuous illumination 12hd
-1, light intensity 20-40umolm
-1S
-1
The sprouting of aseptic seed is to win the blue capsule of uncracked lotus lobe in the above-mentioned method, cleans with 70% ethanol, uses 75% alcohol immersion 20s then, uses 0.2%HgCl again
2Sterilization 30min, aseptic water washing 3 times, each 10min is placed on capsule on the aseptic filter paper, blots fruit surface moisture, cuts off capsule, and seed is evenly broadcast on seed culture medium.
Enrichment culture is 5 months in the said method, root-like stock cutting subculture, subculture 3-4 month.
Embodiment:
Further specify essentiality content of the present invention with embodiments of the invention below, but content of the present invention is not as limit.
Embodiment 1:
Botanical name: lotus lobe orchid (Cymbidium lianpan Tang et Wang)
Material classification: seed
Condition of culture: minimal medium is for spending (No. 1, Hypenox, N: P: K=7: 6: 19, U.S. Hyponex chemical company product) 2.5g+MS organic principle, molysite and trace element+AC 0.5gL precious No. 1
-1+ S 2%.Agar solidifies.
1. the planting seed medium is: minimal medium+BA 5mgL
-1+ NAA 5mgL
-1
2. proliferated culture medium is: minimal medium+BA3mgL
-1+ NAA 5mgL
-1
3. the inducing culture of bud is: minimal medium+BA3mgL
-1+ IAA0.2mgL
-1
4. root media is: minimal medium+NAA1mgL
-1+ banana puree 20%, medium pH 5.5, cultivation temperature (26 ± 2) ℃, continuous illumination 12hd
-1, light intensity 20-40umolm
-1S
-1
Growth and differentiation situation:
1, the sprouting of aseptic seed: win the blue capsule of uncracked lotus lobe,, use 75% alcohol immersion 20s then, use 0.2%HgCl again with dust and the foreign material on the clean fruit of the careful sassafras of gauze that speckles with 70% ethanol
2Sterilization 30min, aseptic water washing 3 times, each 10min.Capsule is placed on the aseptic filter paper, blots fruit surface moisture, cut off capsule, seed is evenly broadcast at medium 1. gone up, the blue seed culture of lotus lobe is about 6 months, and sprouting grows several milky protocorms of 1-, changes green later on; Cultivation through 30d just can differentiate root-like stock again.Seed is under light or dark the cultivation, and sprout time and difference on effect are not obvious.
2, enrichment culture: change root-like stock over to medium and 2. go up cultivation, the rate of increase was 1 in about 5 months: 3-5 (different and different according to kind).These root-like stocks can cut subculture, and subculture is with individual month of about 3-4, and with the increase of subculture number, the rate of increase also increases, and reaches the purpose of quick breeding.
3, inducing of test-tube plantlet: the root-like stock clump changes medium over to and 2. goes up, and it is green that root-like stock changes gradually, has the part bud to differentiate simultaneously, and the agglomerate that will have bud and root-like stock clump is cut into area and is about 1cm
2, changeing and 3. go up as medium, root-like stock differentiates clump bud, and the clump sorite is cut into the 2-3 sprout tuber, is seeded in medium and 4. goes up, and through cultivation in about 3 months, bud grew several roots of 1-, grows tall simultaneously about about 5cm, and rooting rate reaches 98%.
4, acclimatization and transplants:, be placed on 40 μ molm when tissue cultivating seedling grows to 5cm when above
-1S
-1Light intensity lower refining seedling 15-30d.When vane extension, plant grow tall and are healthy and strong, open bottle cap, carefully seedling is taken out, carefully remove root agar, to transplant in the sterile sphagna of moistening but stubborn not water outlet, the lid plastic foil is preserved moisture, and keeps about 25 ℃ of environmental temperature.Will noting in 1 week after moving ventilates preserves moisture, and treats that test-tube plantlet grows young leaves or new root can be removed coverlay, Routine Management.Transplanting survival rate can reach 90%.
Excellent effect of the present invention is: the blue special product of lotus lobe is rare orchid in From Western Yunnan. Its form grace, plant type are beautiful; The blade four seasons are green for a long time; Spend luicd and elegant graceful delicate fragrance, pattern is abundant, has very high ornamental value and economic worth. It is orchid treasure (such as element etc. in slight snow) that many kinds are wherein arranged, and a thousand pieces of gold difficulty is asked. The blue seed of lotus lobe is little, extremely difficult germination, and division propagation is again slow. The clonal foundation of the blue group training seedling of lotus lobe of the present invention is for the breeding of these treasures provides an approach faster. Show through consulting reference materials, cultivate with tissue about the seed asepsis sprouting of lotus lobe orchid and there is not yet so far report.
Claims (2)
1. aseptic seeding of lotus lobe orchid and method for tissue culture are got lotus lobe orchid, and process seed germination, enrichment culture, test-tube plantlet are induced, the acclimatization and transplants step, it is characterized in that winning the blue capsule of uncracked lotus lobe, clean with 70% ethanol, use 75% alcohol immersion 20s then, use 0.2%HgCl again
2Sterilization 30min, aseptic water washing 3 times, each 10min is placed on capsule on the aseptic filter paper, blots fruit surface moisture, cuts off capsule, and seed is broadcast at minimal medium+BA5mgL
-1+ NAA 5mgL
-1Seed culture medium on cultivated 6-7 month, differentiate root-like stock, root-like stock is inserted minimal medium+BA3mgL
-1+ NAA5mgL
-1Proliferated culture medium on carry out enrichment culture, treat that root-like stock changes green gradually, the part bud differentiates, the agglomerate that will have bud and root-like stock clump is cut into 1cm
2Size inserts minimal medium+BA3mgL
-1+ IAA0.2mgL
-1The bud inducing culture on, root-like stock differentiates clump bud, and the clump sorite is cut into the 2-3 sprout tuber, is seeded in minimal medium+NAA1mgL
-1+ banana puree 20%, on the root media of pH5.5 3 months; The minimal medium that relates in the abovementioned steps is N: P: K=7: the product of 6: 19 U.S. Hyponex chemical company is spent precious No. 1 2.5g+MS organic principle, molysite and trace element+AC 0.5gL
-1+ sulphur 2%, agar solidifies; 26 ± 2 ℃ of cultivation temperature, continuous illumination 12hd
-1, intensity of illumination 20-40 μ molm
-2S
-1Treat that bud grows one to several roots, seedling grows to 5cm when above, is placed on 40 μ molm
-2S
-1Light intensity lower refining seedling 15-30 days when vane extension, plant grow tall and be healthy and strong, takes out seedling, transplanting is preserved moisture in moistening sterilization sphagna, and environmental temperature keeps 25 ℃, the lid plastic foil ventilates and preserves moisture, and treats that test-tube plantlet grows young leaves or new root is removed coverlay, Routine Management.
2. the method for claim 1 is characterized in that, root-like stock enrichment culture 5 months, these root-like stock cutting subcultures, subculture 3-4 month.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN2007100659413A CN101066041B (en) | 2007-06-08 | 2007-06-08 | Aseptic seeding and tissue culture process of cymbidium lianpan |
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| CN2007100659413A CN101066041B (en) | 2007-06-08 | 2007-06-08 | Aseptic seeding and tissue culture process of cymbidium lianpan |
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| Publication Number | Publication Date |
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| CN101066041A CN101066041A (en) | 2007-11-07 |
| CN101066041B true CN101066041B (en) | 2011-01-12 |
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| CN2007100659413A Expired - Fee Related CN101066041B (en) | 2007-06-08 | 2007-06-08 | Aseptic seeding and tissue culture process of cymbidium lianpan |
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Families Citing this family (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102379242A (en) * | 2011-08-29 | 2012-03-21 | 张笑逸 | Supersonic wave wall breaking culture method for Chinese terrestrial orchid progenitor cell embryo |
| CN105340757B (en) * | 2015-12-14 | 2017-08-25 | 广东省农业科学院环境园艺研究所 | A kind of method for tissue culture of Cymbidium lianpan and its application |
| CN109122325A (en) * | 2018-11-09 | 2019-01-04 | 翁源县天下泽雨农业科技有限公司 | A kind of aseptic seeding quick-breeding method of sword-leaved cymbidium seed |
| CN109984030B (en) * | 2019-04-18 | 2022-06-17 | 福建农林大学 | Establishment and rapid propagation method of in vitro regeneration system of cymbidium tortisepalum unisexual variety |
| CN112136687B (en) * | 2020-10-28 | 2022-02-18 | 中国科学院昆明植物研究所 | Rapid propagation and in-vitro preservation method of saprophytic orchid |
| CN115589944A (en) * | 2022-08-25 | 2023-01-13 | 云南省农业科学院花卉研究所（Cn） | Method for culturing cymbidium tortisepalum plants on large scale |
Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1535571A (en) * | 2003-04-10 | 2004-10-13 | 中国科学院华南植物研究所 | Sterile sowing and tissue culture method of tree orchid |
| CN1541519A (en) * | 2003-11-06 | 2004-11-03 | 中国科学院华南植物研究所 | Paphiopedilum aseptic seeding and tissue culture technology |
| CN1188029C (en) * | 2003-06-04 | 2005-02-09 | 中国科学院昆明植物研究所 | Induction muthod of externally-introduced lily Casa Balanca test tube bulb |
| CN1799336A (en) * | 2005-12-16 | 2006-07-12 | 江苏阳光生态农林开发股份有限公司 | Method for cultivating cymbidium goeringii seed by tissue culture |
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2007
- 2007-06-08 CN CN2007100659413A patent/CN101066041B/en not_active Expired - Fee Related
Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1535571A (en) * | 2003-04-10 | 2004-10-13 | 中国科学院华南植物研究所 | Sterile sowing and tissue culture method of tree orchid |
| CN1188029C (en) * | 2003-06-04 | 2005-02-09 | 中国科学院昆明植物研究所 | Induction muthod of externally-introduced lily Casa Balanca test tube bulb |
| CN1541519A (en) * | 2003-11-06 | 2004-11-03 | 中国科学院华南植物研究所 | Paphiopedilum aseptic seeding and tissue culture technology |
| CN1799336A (en) * | 2005-12-16 | 2006-07-12 | 江苏阳光生态农林开发股份有限公司 | Method for cultivating cymbidium goeringii seed by tissue culture |
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