CN104429961A - Rapid propagation tissue culture method for dendrobium officinale - Google Patents
Rapid propagation tissue culture method for dendrobium officinale Download PDFInfo
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Abstract
The invention relates to a rapid propagation tissue culture method for dendrobium officinale. The rapid propagation tissue culture method comprises an induced culture stage, a subculture stage and a rooting culture stage, wherein induced culture is carried out by adopting a culture medium which is formed by an improved MS culture medium, 0.5mg/L of 6-BA (6-benzylamino adenine), 5% of banana slurry and 0.1% of active carbon at the induced culture stage; subculture and rooting culture are carried out by adopting a culture medium which is formed by the improved MS culture medium and 0.1mg/L of NAA (naphthylacetic acid); the illumination time in each day is 16 hours, the illumination intensity is 2,000-2500lux, the temperature is controlled to be 25+/-2 DEG C, and the co-culture time is 12-15 weeks at the induced culture stage, the subculture stage and the rooting culture stage. By adopting the culture medium formula and the culture method disclosed by the invention, dendrobium officinale can be efficiently bred; the propagation multiple is over 5n; a nursery stock is healthy in growth; the bacteria and fungi infection rate is low; the transplanting survival rate can be up to over 95%; and industrialized production is achieved.
Description
Technical field:
The present invention relates to technical field of plant cultivation, refer in particular to a kind of Fast-propagation tissue culture method of dendrobium candidum.
Background technology:
Dendrobium candidum (formal name used at school: Dendrobium officinale Kimura et Migo), have another name called: ribbed hedyotis herb, Yunnan iron sheet, belong to micro-specific item, the perennial herbaceous plant that grows nonparasitically upon another plant of the orchid family, is mainly distributed in the ground such as Chinese Anhui, Zhejiang, Fujian.Its stem is used as medicine, and belongs to the yin tonics in tonic: reinforcing stomach reg fluid, nourishing Yin and clearing heat.
At present, candidum tissue culturing seedling growth is comparatively slow, and the Growth of Dendrobium candidum cycle is long, require higher to medium nutrient content, cost is higher, and culturing room's environment is larger on its impact, easy generation germ contamination, fungal contamination, causes candidum tissue culturing seedling survival rate low.Although candidum tissue culturing fast breeding technique has greater advance in recent years, ripe tissue culture technology is efficiently also in testing further and groping.
Summary of the invention:
The object of the invention is to the weak point overcoming existing dendrobium candidum propagation method, a kind of Fast-propagation tissue culture method of dendrobium candidum is provided.
Method of the present invention comprises following steps: a kind of Fast-propagation tissue culture method of dendrobium candidum, comprise Fiber differentiation, squamous subculture and culture of rootage stage, wherein: in the described Fiber differentiation stage, adopt the medium be made up of the 6-BA of modified MS medium, 0.5mg/L (6-benzyl aminoadenine), the banana puree of weight ratio 5%, the active carbon of weight ratio 0.1% to carry out Fiber differentiation; The medium be made up of the NAA of modified MS medium, 0.1mg/L (methyl α-naphthyl acetate) is all adopted to carry out squamous subculture, culture of rootage in described squamous subculture stage and culture of rootage stage.
At Fiber differentiation, squamous subculture and in the culture of rootage stage, illumination every day 16h, intensity of illumination 2000-2500lux, temperature controls at 25 DEG C ± 2 DEG C, 12 weeks-15 weeks of Dual culture.
Obtain a bottle seedling of taking root after described culture of rootage, hardening of uncapping 5 days, re-uses pine bark and transplants as matrix.
In the described Fiber differentiation stage, to the seed disinfection of screening, packing, autoclaving process are carried out to adopted medium; Pathologic finding is carried out to just training seedling in squamous subculture, culture of rootage stage.
Adopt culture medium prescription provided by the invention and cultural method, efficiently can breed dendrobium candidum, expanding numerous multiple is 5
nabove, and seedling growth is healthy and strong, and bacterium, fungal infection rate are low, and transplanting survival rate, up to more than 95%, is conducive to realizing industrialization and produces.
Accompanying drawing illustrates:
Fig. 1 is detailed implementation step block diagram of the present invention.
Embodiment:
Below in conjunction with specific embodiments and the drawings, the present invention is further described.
The Fast-propagation tissue culture method of a kind of dendrobium candidum of the present invention, it comprises Fiber differentiation, squamous subculture and culture of rootage stage, and detailed step as shown in Figure 1.
Improvements of the present invention are: adopt the medium be made up of the banana puree of the 6-BA of modified MS medium, 0.5mg/L, weight ratio 5%, the active carbon of weight ratio 0.1% to carry out Fiber differentiation in the described Fiber differentiation stage; The medium be made up of the NAA of modified MS medium, 0.1mg/L is all adopted to carry out squamous subculture, culture of rootage in described squamous subculture stage and culture of rootage stage.
At Fiber differentiation, squamous subculture and in the culture of rootage stage, illumination every day 16h, intensity of illumination 2000-2500lux, temperature controls at 25 DEG C ± 2 DEG C, 12 weeks-15 weeks of Dual culture.
Obtain a bottle seedling of taking root after described culture of rootage, hardening of uncapping 5 days, re-uses pine bark and transplants as matrix.
In the described Fiber differentiation stage, to the seed disinfection of screening, packing, autoclaving process are carried out to adopted medium; Pathologic finding is carried out to just training seedling in squamous subculture, culture of rootage stage.
Adopt culture medium prescription provided by the invention and cultural method, efficiently can breed dendrobium candidum, expanding numerous multiple is 5
nabove, and seedling growth is healthy and strong, and bacterium, fungal infection rate are low, and transplanting survival rate, up to more than 95%, is conducive to realizing industrialization and produces.
Claims (4)
1. the Fast-propagation tissue culture method of a dendrobium candidum, comprise Fiber differentiation, squamous subculture and culture of rootage stage, it is characterized in that: in the described Fiber differentiation stage, adopt the medium be made up of the banana puree of the 6-BA of modified MS medium, 0.5mg/L, weight ratio 5%, the active carbon of weight ratio 0.1% to carry out Fiber differentiation; The medium be made up of the NAA of modified MS medium, 0.1mg/L is all adopted to carry out squamous subculture, culture of rootage in described squamous subculture stage and culture of rootage stage.
2. the Fast-propagation tissue culture method of dendrobium candidum according to claim 1, it is characterized in that: at Fiber differentiation, squamous subculture and in the culture of rootage stage, illumination every day 16h, intensity of illumination 2000-2500lux, temperature controls at 25 DEG C ± 2 DEG C, 12 weeks-15 weeks of Dual culture.
3. the Fast-propagation tissue culture method of dendrobium candidum according to claim 1 and 2, is characterized in that: obtain a bottle seedling of taking root after described culture of rootage, hardening of uncapping 5 days, re-uses pine bark and transplant as matrix.
4. the Fast-propagation tissue culture method of dendrobium candidum according to claim 1 and 2, is characterized in that: in the described Fiber differentiation stage, to the seed disinfection of screening, carries out packing, autoclaving process to adopted medium; Pathologic finding is carried out to just training seedling in squamous subculture, culture of rootage stage.
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| CN201410741417.3A CN104429961B (en) | 2014-12-08 | 2014-12-08 | A kind of Fast-propagation tissue culture method of Herba Dendrobii |
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| CN201410741417.3A CN104429961B (en) | 2014-12-08 | 2014-12-08 | A kind of Fast-propagation tissue culture method of Herba Dendrobii |
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| CN104429961A true CN104429961A (en) | 2015-03-25 |
| CN104429961B CN104429961B (en) | 2016-08-24 |
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| CN104823853A (en) * | 2015-05-11 | 2015-08-12 | 安徽康尔欣生物制药有限公司 | Method for rapidly proliferating tissue culture sprout of dendrobium |
| CN108617511A (en) * | 2018-08-10 | 2018-10-09 | 浙江省东阳玉米研究所 | The seed group fast tissue culture reproducing method of dendrobium candidum |
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| CN104429961B (en) | 2016-08-24 |
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