CN102696485A - Fast propagation method of dharma orchid - Google Patents
Fast propagation method of dharma orchid Download PDFInfo
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- CN102696485A CN102696485A CN2012102214608A CN201210221460A CN102696485A CN 102696485 A CN102696485 A CN 102696485A CN 2012102214608 A CN2012102214608 A CN 2012102214608A CN 201210221460 A CN201210221460 A CN 201210221460A CN 102696485 A CN102696485 A CN 102696485A
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Abstract
The invention provides a fast propagation method of dharma orchid, belonging to the technique of regeneration of dharma orchid plants and industrialized fast production of the dharma orchid. With the adoption of the fast propagation method, the problems of lack stock plants, less propagation base, low propagation rate, slow differentiating speed, small seeds, slow breeding progress and low emergence rate are solved. The fast propagation method comprises the following steps: selecting a strong stock plant of an original seedling of the dharma orchid without pest and disease damage and cutting 3-5cm of a lateral bud from the strong stock plant as an explant material; after carrying out stem tip induction culture on the stock plant for 20 days, growing many white granular calluses around the stock plant; continuously culturing to change the white granular calluses into green calluses and then forming a lot of corms after 25 days; carrying out adventitious bud induction by the differentiating a culture medium and differentiating after 30-40 days to gradually generate cluster buds; under the condition that the cluster buds reaches certain quantity, carrying out rooting culture; and after rooting, acclimatizing and transplanting, thereby the propagation coefficient is expanded, the breeding cycle is shortened, the market demand is satisfied, and a technical support for industrialized propagation of a large quantity of rare dharma orchid is provided.
Description
Technical field
The invention belongs to Bodhidharma blue plant regeneration and the quick production technology of industrialization, relate to the blue method for quickly breeding of a kind of Bodhidharma.
Background technology
The Bodhidharma orchid is a kind of of Chinese cymbidium, and the fragrance of a flower is pure deep and remote far away, spends 7～9, and red wish is a kind of ornamental plants of preciousness in 1～March of florescence.But therefore difficult problems such as the blue existing production technology of Bodhidharma exists maternal plant rare at present, and the breeding radix is few, and reproduction rate is low, and plant division speed is slow, and seed is little, and breeding process is slow, and emergence rate is low, are difficult to obtain desirable germination rate and the strong rare seedling of more key.
Summary of the invention
The object of the invention provides the blue method for quickly breeding of a kind of Bodhidharma; The healthy and strong unanimity of the seedling that this method is cultivated, generation time weak point, one-tenth seedling are transplanted back survival rate height, and cost is low, the florescence unanimity; Be convenient to the control of industrialization unified management and selling time, have remarkable economic efficiency.
The technical scheme that the present invention takes is following:
The blue method for quickly breeding of a kind of Bodhidharma, concrete steps comprise:
1) draws materials, sterilizes: choose the blue former seedling of Bodhidharma, cut the lateral bud of 3～5 centimetres of length, after rinsing well; In water, cut off outer 1～2 blade tip of root and lateral bud, clean, disinfect, peel off peripheral organization; Cut stem apex, directly be inoculated on the protocorm inducing culture;
2) protocorm is induced: the prescription of said protocorm inducing culture is: add 6-BA 0.5mg/L among the minimal medium MS; NAA 1.0 mg/L; Banana pulp 150 g/L; Peptone 3g/L; White sugar 2%; Active carbon 2g/L; Agar 6.5g, condition of culture: culturing room's temperature is: 25 ± 2 ℃, light application time 12h/d, intensity of illumination 1000～1500Lx induces the generation protocorm;
3) inducing clumping bud: choose step 2) well-grown protocorm forwards on the inducing clumping bud medium, and the prescription of said inducing clumping bud medium is: add 6-BA 3.0mg/L among minimal medium 1/2 MS; NAA 1.0 mg/L; Peptone 2g/L; Spend precious No. 12 g/L; Active carbon 2.0 g/L; White sugar 3%; Banana pulp 150 g/L; Condition of culture: culturing room's temperature is: 25 ± 2 ℃, and light application time 12h/d, intensity of illumination 1000～1500Lx; Induce the bud of growing thickly;
4) enrichment culture: will cross step 3) and induce the bud of growing thickly of generation to cut, and be inoculated in the proliferated culture medium; 3 of the proliferated culture medium inoculations of per 100～150g; The prescription of described proliferated culture medium is: add 6-BA 0.5mg/L among minimal medium 1/2 MS; NAA 0.5mg/L; Active carbon 2.0 g/L; Spend precious No. 22 g/; White sugar 3%; Banana pulp 150 g/L; Condition of culture: culturing room's temperature is: 25 ± 2 ℃, and light application time 12h/d, intensity of illumination 1000～1500Lx;
5) root induction: the seedling individual plant that grows to 2cm height, 3～4 leaves that the step 4) enrichment culture is come out downcuts, and is transferred on the root media, and the prescription of described root media is: add 6-BA 0.3mg/L among minimal medium 1/2 MS; NAA 0.5 mg/L; IBA 1.0 mg/L; Active carbon 2.0 g/L; White sugar 3%; Condition of culture: culturing room's temperature is: 25 ± 2 ℃, and light application time 12h/d, intensity of illumination 1000～1500Lx;
6) test-tube plantlet is accomplished: when test-tube plantlet grows to 4～5cm, when 3～4 roots of tool, 4～5 leaves, accomplish the blue test-tube plantlet of Bodhidharma and cultivate;
The above minimal medium 1/2 MS refers to that macroelement reduces by half.
The blue test-tube plantlet of Bodhidharma that said step 6) is accomplished is transplanted, and the natural conditions lower refining seedling 4～5d outside culturing room of elder generation before transplanting opened bottle cap refining seedling 2～4 days then; From bottle, take out seedling behind the refining seedling; Clean the medium that adheres to the seedling root with clear water, root is wrapped, transplant to the mixed-matrix of perlite, microlith and bark with sphagna; And bind up with gauze 3～5 slow-release fertilizers and place the transplanting dish, note not being placed directly in root.
Wherein spend precious No. 1 composition ratio: the N:P:K proportioning is: (7:6:19), N, P, K refers to respectively: NO
3Nitrate nitrogen, PO
3Phosphorous oxide, K0
3Potassium oxide.
The healthy and strong unanimity of the seedling that the present invention cultivates, generation time weak point, one-tenth seedling are transplanted back survival rate height, and cost is low, and the florescence unanimity is convenient to the control of industrialization unified management and selling time, has remarkable economic efficiency.
Description of drawings
Fig. 1 induces the protocorm of formation for stem apex.
Fig. 2 induces the indefinite bud of formation for protocorm.
Fig. 3 induces the bud of growing thickly of formation for the propagation seedling.
Fig. 4 is the Bodhidharma orchid seedling of taking root.
Fig. 5 is the test tube transplanted seedling.
Embodiment
Below in conjunction with embodiment the present invention is described further, but the present invention is not limited only to this.
Embodiment 1
It is explant material that the present invention chooses the lateral bud that cuts 3～5 centimetres of length on the healthy and strong maternal plant of the anosis insect pest of the blue former seedling of Bodhidharma; Through the stem apex inducing culture; The 20d Later Zhou Dynasty, one of the Five Dynasties crosses many at present white granular callus, continues to cultivate to transfer green gradually to, can form a large amount of protocorms after 25 days; Carry out adventitious bud inducing through differential medium again, 30～40d begins differentiation, progressively induces the bud of growing thickly, and breed after some and carry out culture of rootage again, but acclimatization and transplants after taking root.Thereby the expanding propagation coefficient shortens growing-seedling period, meets the need of market, and carrying out industrialization production for the blue a large amount of breedings of rare species Bodhidharma provides technical support.
Medium:
The MS minimal medium adopts 1986, and Beijing Higher Education Publishing House publishes, and the prescription in Chen Zhenghua chief editor " woody plant tissure is cultivated and used " is prepared.
The protocorm inducing culture is: add 6-BA 0.5mg/L among the minimal medium MS; NAA 1.0 mg/L; Banana pulp 150 g/L; Peptone 3g/L; White sugar 2%; Active carbon 2g/L; Agar 6.5g.
The inducing clumping bud medium is: add 6-BA 3.0mg/L among minimal medium 1/2 MS (macroelement reduces by half); NAA 1.0 mg/L; AC 2.0g/L; Banana meat 150 g/L; Peptone 2g/L; Spend precious No. 12 g/L; AC 2.0 g/L; White sugar 3%.
Proliferated culture medium is: add among minimal medium 1/2 MS (macroelement reduces by half); 6-BA 0.5mg/L; NAA 0.5mg/L; AC 2.0 g/L; Spend precious No. 22 g/; White sugar 3%; Banana meat 150 g/L.
Root media is: add 6-BA 0.3mg/L among minimal medium 1/2 MS (macroelement reduces by half); NAA 0.5 mg/L; IBA 1.0 mg/L; AC 2.0 g/L; White sugar 3%.
Minimal medium 1/2MS is meant: macroelement reduces by half in the MS minimal medium, other components unchanged.
Materials disinfection is handled: earlier material is suitably cleaned, with the long stem segment with axillary bud of scissors intercepting 2～3cm, in water, cut off outer 1～2 blade tip of root and lateral bud; Use the washing powder clean surface, about flowing water flushing 30min, place on the superclean bench; With 75% alcohol-pickled 3min, then with 0.1% mercuric chloride sterilization 8min, behind the aseptic water washing 3～4 times with rifampin (70～75mg/L) immersion 3min; And with aseptic water washing 1 time; Peel off peripheral organization, cut stem apex, directly be inoculated on the protocorm inducing culture.
Protocorm is induced: condition of culture: culturing room's temperature is: 25 ± 2 ℃, and light application time 12h/d, intensity of illumination 1000～1500Lx; Induce about 25d, as shown in Figure 1, on average induce 7～8 of protocorms, inductivity reaches more than 96%.
Inducing clumping bud: with said through step 2) induce the protocorm of generation to repeat to cultivate after; Choosing well-grown protocorm forwards on the inducing clumping bud medium; Described condition of culture: culturing room's temperature is: 25 ± 2 ℃, and light application time 12h/d, intensity of illumination 1000～1500Lx; Begin differentiation about 40d, induce the bud of growing thickly, induce differentiation rate high like Fig. 2 bud, stalwartness, dark green.
Enrichment culture: induce the bud of growing thickly of generation to cut said through step 3), be seeded in the proliferated culture medium respectively; 3 of the proliferated culture medium inoculations of 100～150g; Described condition of culture: culturing room's temperature is: 25 ± 2 ℃, and light application time 12h/d, intensity of illumination 1000～1500Lx, as shown in Figure 4, the bud of growing thickly is sprouted and is formed a large amount of plant, and bud is healthy and strong, and the rate of increase is high.
Root induction: the said seedling individual plant of cultivating out through step 4) that grows to 2cm height, 3～4 leaves is downcut; Be transferred on the different root medias, every bottle 5 strain, described condition of culture: culturing room's temperature is: 25 ± 2 ℃; Light application time 12h/d; Intensity of illumination 1000～1500Lx, rooting rate reach more than 90%, and be as shown in Figure 4.
Test-tube plantlet is accomplished: when test-tube plantlet grows to 4～5cm, during 3～4 roots of tool, during 4～5 leaves, accomplishes blue plant regeneration of Bodhidharma and industrialization production key technology test-tube plantlet and cultivates, and as shown in Figure 5.
The blue test-tube plantlet of the Bodhidharma that said step 6) is accomplished is transplanted, and outside culturing room, refines seedling about one week earlier before transplanting.From bottle, take out then, clean the medium that adheres to, root is wrapped with sphagna; Transplant to the mixed-matrix of perlite, microlith and bark; And bind up with gauze 3～5 slow-release fertilizers and place the transplanting dish, note not being placed directly in root, transplant the back and keep certain temperature and humidity.
Claims (2)
1. the blue method for quickly breeding of a Bodhidharma, it is characterized in that: the concrete steps of said method comprise:
1) draws materials, sterilizes: choose the blue former seedling of Bodhidharma, cut the lateral bud of 3～5 centimetres of length, after rinsing well; In water, cut off outer 1～2 blade tip of root and lateral bud, clean, disinfect, peel off peripheral organization; Cut stem apex, directly be inoculated on the protocorm inducing culture;
2) protocorm is induced: the prescription of said protocorm inducing culture is: add 6-BA 0.5mg/L among the minimal medium MS; NAA 1.0 mg/L; Banana pulp 150 g/L; Peptone 3g/L; White sugar 2%; Active carbon 2g/L; Agar 6.5g, condition of culture: culturing room's temperature is: 25 ± 2 ℃, light application time 12h/d, intensity of illumination 1000～1500Lx induces the generation protocorm;
3) inducing clumping bud: choose step 2) well-grown protocorm forwards on the inducing clumping bud medium, and the prescription of said inducing clumping bud medium is: add 6-BA 3.0mg/L among minimal medium 1/2 MS; NAA 1.0 mg/L; Peptone 2g/L; Spend precious No. 12 g/L; Active carbon 2.0 g/L; White sugar 3%; Banana pulp 150 g/L; Condition of culture: culturing room's temperature is: 25 ± 2 ℃, and light application time 12h/d, intensity of illumination 1000～1500Lx; Induce the bud of growing thickly;
4) enrichment culture: will cross step 3) and induce the bud of growing thickly of generation to cut, and be inoculated in the proliferated culture medium; 3 of the proliferated culture medium inoculations of per 100～150g; The prescription of described proliferated culture medium is: add 6-BA 0.5mg/L among minimal medium 1/2 MS; NAA 0.5mg/L; Active carbon 2.0 g/L; Spend precious No. 22 g/; White sugar 3%; Banana pulp 150 g/L; Condition of culture: culturing room's temperature is: 25 ± 2 ℃, and light application time 12h/d, intensity of illumination 1000～1500Lx;
5) root induction: the seedling individual plant that grows to 2cm height, 3～4 leaves that the step 4) enrichment culture is come out downcuts, and is transferred on the root media, and the prescription of described root media is: add 6-BA 0.3mg/L among minimal medium 1/2 MS; NAA 0.5 mg/L; IBA 1.0 mg/L; Active carbon 2.0 g/L; White sugar 3%; Condition of culture: culturing room's temperature is: 25 ± 2 ℃, and light application time 12h/d, intensity of illumination 1000～1500Lx;
6) test-tube plantlet is accomplished: when test-tube plantlet grows to 4～5cm, when 3～4 roots of tool, 4～5 leaves, accomplish the blue test-tube plantlet of Bodhidharma and cultivate;
The above minimal medium 1/2 MS refers to that macroelement reduces by half.
2. the blue method for quickly breeding of Bodhidharma according to claim 1; It is characterized in that: the blue test-tube plantlet of Bodhidharma that said step 6) is accomplished is transplanted, and the natural conditions lower refining seedling 4～5d outside culturing room of elder generation before transplanting opened bottle cap refining seedling 2～4 days then; From bottle, take out seedling behind the refining seedling; Clean the medium that adheres to the seedling root with clear water, root is wrapped, transplant to the mixed-matrix of perlite, microlith and bark with sphagna; And bind up with gauze 3～5 slow-release fertilizers and place the transplanting dish, note not being placed directly in root.
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Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104782368A (en) * | 2015-04-16 | 2015-07-22 | 济宁森立生物科技有限公司 | Saffron crocus corm propagation method |
| CN105638466A (en) * | 2016-01-05 | 2016-06-08 | 刘华武 | Fragrant thoroughwort cultivation method |
| CN106417012A (en) * | 2016-08-30 | 2017-02-22 | 贵州德江易盛农业科技发展有限公司 | Tissue culture method for promoting efficient propagation of Thunia alba |
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Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104782368A (en) * | 2015-04-16 | 2015-07-22 | 济宁森立生物科技有限公司 | Saffron crocus corm propagation method |
| CN105638466A (en) * | 2016-01-05 | 2016-06-08 | 刘华武 | Fragrant thoroughwort cultivation method |
| CN106417012A (en) * | 2016-08-30 | 2017-02-22 | 贵州德江易盛农业科技发展有限公司 | Tissue culture method for promoting efficient propagation of Thunia alba |
| CN106417012B (en) * | 2016-08-30 | 2018-12-18 | 贵州德江易盛农业科技发展有限公司 | A kind of tissue culture method for promoting bamboo shoot orchid efficiently to breed |
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Application publication date: 20121003 |