CN1729747A - Cymbidium plant seed asepsis sprouting and plant cultivating method - Google Patents

Abstract

The invention discloses a method of sterile germination of seed and plant cultivating for thoroughwort. Collecting Cmbidium tracyanum L castle plants, C. iridioides plants and jasper orchid plants and cultivating them in the resource garden until the fruit of the orchid is 90% mature. The cultivating method compeises: culturing seeds in the modified culture medium by explant, trans-culturing when the seed grows sprout and root with root hair, rapid breeding and root-growing culturing, new root will grow out one month later. The traditional orchid breeding is primarily the single-plant propagation with a low propagation rate, the portion between old sprout and mew sprout is only about 1:1-3,and is hard for mass production. The invention applies modern biotechnology and breed millions of orchids only employing one sprout in one year, which can meet the orchid industrialization demand; sterilizing technique can also be proceeded and produce seeds free of virus, which deduces the production cost, increase the product quality and meet the orchid industrialization demand effectively.

Description

Cymbidium plant seed asepsis sprouting and plant cultivating method

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, the blue strain of yellow cicada, 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 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 1/2MS+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, the blue strain of yellow cicada, 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 (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 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 1/2MS+10 to from base portion ^-6Y ₂Or 10 ^-6Y ₁In, promptly grow new root after one month.