CN100358413C - Cymbidium plant seed asepsis sprouting and plant cultivating method - Google Patents
Cymbidium plant seed asepsis sprouting and plant cultivating method Download PDFInfo
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- CN100358413C CN100358413C CNB200510010764XA CN200510010764A CN100358413C CN 100358413 C CN100358413 C CN 100358413C CN B200510010764X A CNB200510010764X A CN B200510010764XA CN 200510010764 A CN200510010764 A CN 200510010764A CN 100358413 C CN100358413 C CN 100358413C
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Abstract
The present invention relates to a method for the sterile germination of cymbidium plant seeds and plant culture. Cymbidium tracyanum L castle plants, C. Lowianum plants and C. iridioides plants are cultured in a resource garden until orchid fruits are 90% mature. The culture method comprises the following procedures: explants are processed, and seeds are cultured by improved culture media; when the seeds are germinated and small roots with root hair grow from growth buds, the buds are transplanted and cultured; the buds are rapidly propagated, rooted and cultured; new roots grow from seedlings after one month. Traditional orchid propagation mainly takes single plant propagation as the main, but the coefficient of the single plant propagation is low. The proportion of old seedlings to new seedlings is only 1:1 to 3 more or less every year, and mass production can not be realized by the single plant propagation. In the present invention, a plurality of hundred thousand orchid plants can be propagated from one bud in one year by a modern biologic technique, so requirements for orchid industrial production are met. Operation by detoxification technology can also be carried out for producing seedlings without viruses, so orchid production cost can be reduced, product quality can be improved, and large requirements for orchid industrial production can be effectively met.
Description
Technical field
The present invention relates to the domestication of plants technical field, specifically cymbidium plant seed asepsis sprouting and plant cultivating method.
Background technology
Cymbidium iridioides (Cmbidium tracyanum L castle), Cymbidium hookerianum var. Lowianum (Rchb.f.)Y.S.wu et S.C.chen (C.Lowianum) and yellow cicada orchid (C.iridioides) are the orchid family epidendrums, and the flower bough is long, and the ornamental value height is excellent orchid resource.Three kinds of orchid fecund in Yunnan, Guizhou and the southeast, Tibet, be good hybrid strain and cut-flower material.
Cymbidium seed is very little, includes the globular embryo of some ateliosis, and no endosperm be difficult to be sprouted under the nature, needs and the achieving success that cultivates one's ability of blue bacterium symbiosis or aseptic condition.The sprouting of Cymbidium (Cmbidium) plant seed, existing people did research, but the axenic germination of above three kinds of epidendrums does not appear in the newspapers.
Summary of the invention
The purpose of this invention is to provide a kind of cymbidium plant seed asepsis sprouting and plant cultivating method.
The present invention has carried out axenic germination and quick propagating technology architectural study to Cymbidium iridioides (Cmbidium tracyanum L castle), Cymbidium hookerianum var. Lowianum (Rchb.f.)Y.S.wu et S.C.chen (C.Lowianum) and three kinds of cymbidium seeds of yellow cicada orchid (C.iridioides); successfully cultivate a large amount of tissue cultivating seedling, form crossbreeding and large-scale production and cultivate indispensable key technology.
Concrete grammar of the present invention is:
1) material source: adopt to such an extent that the blue strain of Cymbidium iridioides or the blue strain of yellow cicada or the blue strain of Cymbidium hookerianum var. Lowianum (Rchb.f.)Y.S.wu et S.C.chen are cultivated to orchid fruit to ninety percent ripely in the resource garden, win during pericarp summary displaing yellow;
2) cultural method:
1. explant is handled: with above-mentioned 90% maturity fruit with 75% alcohol disinfecting 40 seconds, change 1% mercuric chloride surface sterilization 8～12 minutes over to, aseptic water washing 3～4 times, desinfection chamber with a small amount of sterile water rinse 5 times, is used 0.1mol/L KOH solution preliminary treatment 8 minutes after taking out seed and binding up with gauze seed then, with aseptic water washing 3 times, in saturated bleaching powder supernatant, sterilized 10～20 minutes, behind the aseptic water washing 5 times, be inoculated into solid culture medium;
2. medium: the MS solid culture medium (being the 1/2MS medium) with improvement adds the basic element of cell division, and growth hormone, i.e. 1/2MS+6-BA1.0～2.0mg.L
-1+ NAA0.5～1.0mg.L
-1Medium is seeded in seed on this medium and cultivates;
3. condition of culture: in whole incubation, seed asepsis sprouting adopts camera bellows to cultivate, cultivation, propagation and the culture of rootage of ball stem Cheng Miao all adopt illumination cultivation, condition of culture is: 25 ± 2 ℃ of temperature, illumination 1800～2500 luxs (lx), illumination every day 12～15 hours, medium pH 5.4, agar 7g/L, when treating to grow bud behind the seed germination and growing the young root of band root hair, switching is cultivated;
4. switching: the ball stem is transferred, promptly forward another blake bottle to from a blake bottle;
5. fast breeding: when ball stem length to about the 1cm, forward proliferated culture medium 1/2 MS+6-BA1.5～2.5mg.L again to
-1+ NAA0.2～0.5mg.L
-1+ banana puree 70～80g/L carries out enrichment culture in cultivating;
6. culture of rootage: the seedling behind the enrichment culture is cultivated 5～8cm height, downcut, forward root media 1/2MS+10 to from base portion
-6Y
2Or 10
-6Y
1In, promptly grow new root after one month.
The present invention draws from experimental study as drawing a conclusion:
1, seed asepsis sprouting is cultivated: the 1/2MS medium adds 6-BA1.0～2.0mg.L
-1+ NAA0.5～1.0mg.L
-1Help seed germination.
2, the ball stem is cultivated: the best results of the medium culture ball stem growth of improvement.
3, different hormones, the influence of medium: best according to growth hormone, the basic element of cell division and other added substances (banana puree) enrichment culture effect that different cultivation periods addings are different to breeding.
4, culture of rootage: the blue seedling of unrooted is transferred in the root media of different activities material, and the short root effect difference of the different activities material when cultivating three months adds 10 at minimal medium
-6Y
2When (biocontrol microorganisms), root length, radical, the thick effect of stem are all good than other three groups, and radical contrasts high by 33%; The contrast that does not add hormone also grows three roots, illustrates the degree of dependence of exogenous hormone not highly, and the growth to root when simultaneously NAA is 2.0mg/L has inhibitory action; Add 10
-6Y
1When (biocontrol microorganisms), root is long, root thick, plant height, stem slightly all contrast.Illustrate that culture of rootage is with 10
-6Y
2(biocontrol microorganisms) is advisable.
The advantage that the present invention has compared with the prior art
1, traditional orchid breeds mainly based on division propagation, and at the bottom of the division propagation coefficient, the ratio of annual Lao Miao and newborn seedling only is about 1: 1～3, is difficult to carry out large-scale production.It is less that tissue culture is applied to the cultivation report of orchid, and this modern biotechnology can breed hundreds thousand of strain orchids with a bud in 1 year, thereby satisfied the orchid industry demand; Also can carry out detoxification technology operation, produce virus-free seedling, thereby can reduce the cost of orchid production and improve the quality of products, this all traditional plant division technology can't be accomplished.Therefore, this technology has overcome all drawbacks that orchid tradition propagation method is brought, and helps enlarging the reproduction coefficient of orchid, can produce the test tube seedling and the detoxic seedling of enormous amount in a short time, effectively satisfies the great demand of orchid industry production.
Embodiment
Embodiment 1:
Material source: adopt from the mountain area, Yunnan the blue strain of Cymbidium iridioides, in the resource garden, cultivate ripely to orchid fruit to ninety percent, pericarp is slightly won during displaing yellow.
Cultural method:
1, explant is handled: with above-mentioned 90% maturity fruit with 75% alcohol disinfecting 40 seconds, change 1% mercuric chloride surface sterilization 8～12 minutes over to, aseptic water washing 3～4 times, desinfection chamber with a small amount of sterile water rinse 5 times, is used 0.1mol/L KOH (potassium hydroxide) solution preliminary treatment 8 minutes after taking out seed and binding up with gauze seed then, with aseptic water washing 3 times, in saturated bleaching powder supernatant, sterilized 10～20 minutes, behind the aseptic water washing 5 times, be inoculated into solid culture medium;
2, medium: use 1/2MS+6-BA1.0mg.L
-1+ NAA1.0mg.L
-1Medium is seeded in seed and cultivates (6-BA is a 6-benzyladenine, is the basic element of cell division, and NAA is a naa, is growth hormone) on the MS medium of this improvement;
3, condition of culture: in whole incubation, except seed asepsis sprouting adopts the camera bellows cultivation, the cultivation of ball stem Cheng Miao, propagation and culture of rootage all adopt illumination cultivation.Condition of culture is: 25 ± 2 ℃ of temperature, illumination 1800～2500 luxs (lx), illumination every day 12～15 hours, medium pH 5.4, agar 7g/L.Inoculate that germination rate reaches more than 90% after 3 months.Grow bud behind the seed germination earlier, grow the young root of band root hair subsequently, Cheng Miao is gradually cultivated after 3 months in switching;
4, Zhuan Jie detailed process: forwarding method: because cymbidium seed is very tiny, some seed flocks together during sowing, causes the round stem ball that grows more crowded, therefore the ball stem is transferred, and promptly forwards the process of another blake bottle to from a blake bottle;
5, fast breeding process: when ball stem length to about the 1cm, forward proliferated culture medium 1/2MS+6-BA2.5mg.L again to
-1+ NAA0.2mgL
-1+ banana puree 70g/L cultivates (banana puree is an additives);
6, culture of rootage: the seedling behind the enrichment culture is cultivated 5～8cm height, downcut, forward root media 1/2MS+10 to from base portion
-6Y
2Or 10
-6Y
1In (Y
1, Y
2Be biocontrol microorganisms, provide by Yunnan Prov Agriculture University), promptly grow new root after one month.
Embodiment 2:
Material source: adopt from the mountain area, Yunnan the blue strain of Cymbidium hookerianum var. Lowianum (Rchb.f.)Y.S.wu et S.C.chen (C.Lowianum), in the resource garden, cultivate ripely to orchid fruit to ninety percent, pericarp is slightly won during displaing yellow.
Cultural method:
1, explant is handled and is undertaken by the method for embodiment 1.
2, medium: use 1/2MS+6-BA2.0mg.L
-1+ NAA0.5mg.L
-1Medium is seeded in seed on the MS medium of this improvement and cultivates;
Step 3 and 4 is undertaken by the method for embodiment 1.
5, fast breeding process: when ball stem length to about the 1cm, forward proliferated culture medium 1/2MS+6-BA1.5mg.L again to
-1+ NAA0.5mg.L
-1+ banana puree 80g/L cultivates;
6 steps are undertaken by the method for embodiment 1.
Embodiment 3:
Material source: adopt from the mountain area, Yunnan the blue strain of yellow cicada orchid (C.iridioides), in the resource garden, cultivate ripely to orchid fruit to ninety percent, pericarp is slightly won during displaing yellow.
Cultural method is undertaken by the method for embodiment 1 or embodiment 2.
Claims (1)
1. cymbidium plant seed asepsis sprouting and plant cultivating method is characterized in that carrying out according to the following steps:
1) material source: adopt to such an extent that the blue strain of Cymbidium iridioides or the blue strain of yellow cicada or the blue strain of Cymbidium hookerianum var. Lowianum (Rchb.f.)Y.S.wu et S.C.chen are cultivated to orchid fruit to ninety percent ripely in the resource garden, win during pericarp summary displaing yellow;
2) cultural method:
1. explant is handled: with above-mentioned 90% maturity fruit with 75% alcohol disinfecting 40 seconds, change 1% mercuric chloride surface sterilization 8～12 minutes over to, aseptic water washing 3～4 times, desinfection chamber with a small amount of sterile water rinse 5 times, is used 0.1mol/L KOH solution preliminary treatment 8 minutes after taking out seed and binding up with gauze seed then, with aseptic water washing 3 times, in saturated bleaching powder supernatant, sterilized 10～20 minutes, behind the aseptic water washing 5 times, be inoculated into solid culture medium;
2. medium: with MS+6-BA1.0～2.0mg.L
-1+ NAA0.5～1.0mg.L
-1Medium is seeded in seed on this medium and cultivates;
3. condition of culture: in whole incubation, seed asepsis sprouting adopts camera bellows to cultivate, cultivation, propagation and the culture of rootage of ball stem Cheng Miao all adopt illumination cultivation, condition of culture is: 25 ± 2 ℃ of temperature, illumination 1800～2500 luxs (1x), illumination every day 12～15 hours, medium pH 5.4, agar 7g/L, when treating to grow bud behind the seed germination and growing the young root of band root hair, switching is cultivated;
4. switching: the ball stem is transferred, promptly forward another blake bottle to from a blake bottle;
5. fast breeding: when ball stem length to about the 1cm, forward proliferated culture medium 1/2MS+6-BA1.5～2.5mg.L again to
-1+ NAA0.2～0.5mg.L
-1+ banana puree 70～80g/L carries out enrichment culture in cultivating;
6. culture of rootage: the seedling behind the enrichment culture is cultivated 5～8cm height, downcut, forward root media 1/2 MS+10 to from base portion
-6Y
2Or 10
-6Y
1In, promptly grow new root after one month.
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|---|---|---|---|
| CNB200510010764XA CN100358413C (en) | 2005-04-22 | 2005-04-22 | Cymbidium plant seed asepsis sprouting and plant cultivating method |
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|---|---|---|---|
| CNB200510010764XA CN100358413C (en) | 2005-04-22 | 2005-04-22 | Cymbidium plant seed asepsis sprouting and plant cultivating method |
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| CN1729747A CN1729747A (en) | 2006-02-08 |
| CN100358413C true CN100358413C (en) | 2008-01-02 |
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Families Citing this family (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103314862B (en) * | 2013-07-16 | 2015-12-02 | 深圳市夺天工园林建设有限公司 | A kind of method of efficient acquisition Chunlan detoxification seedling |
| CN104429863B (en) * | 2014-12-17 | 2017-05-31 | 湖南省核农学与航天育种研究所 | A kind of cultural method of orchid |
| CN108271693B (en) * | 2018-02-11 | 2021-06-01 | 福建农林大学 | Method for quickly establishing and storing regeneration system of parasitic orchid |
| CN109105233A (en) * | 2018-08-08 | 2019-01-01 | 山东省果树研究所 | The copper coin tree seedling fostering method and culture medium quickly bred based on micro cuttage |
| CN115537346B (en) * | 2022-11-23 | 2023-03-24 | 中国科学院昆明植物研究所 | Mucillus mucilaginosus for promoting growth and differentiation of sansevieria trifasciata and application thereof |
Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1335053A (en) * | 2001-08-29 | 2002-02-13 | 中国科学院昆明植物研究所 | Method of bacteria-free germination and fast propagation of peltate yam |
| CN1593107A (en) * | 2004-07-09 | 2005-03-16 | 中国科学院昆明植物研究所 | Yellow moccasin flower seed preparation and quick screening method |
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2005
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Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1335053A (en) * | 2001-08-29 | 2002-02-13 | 中国科学院昆明植物研究所 | Method of bacteria-free germination and fast propagation of peltate yam |
| CN1593107A (en) * | 2004-07-09 | 2005-03-16 | 中国科学院昆明植物研究所 | Yellow moccasin flower seed preparation and quick screening method |
Non-Patent Citations (3)
| Title |
|---|
| 影响杏黄兜兰种子萌发的因素. 丁长春.云南植物研究，第26卷第6期. 2004 * |
| 蝴蝶兰无菌播种技术. 姚丽娟等.北方园艺，第4期. 2004 * |
| 蝴蝶兰无菌萌发技术的研究. 章玉平等.武汉植物学研究，第22卷第1期. 2004 * |
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