CN106718909B - A kind of fast breeding technique of dendrobium candidum - Google Patents
A kind of fast breeding technique of dendrobium candidum Download PDFInfo
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- CN106718909B CN106718909B CN201611177306.XA CN201611177306A CN106718909B CN 106718909 B CN106718909 B CN 106718909B CN 201611177306 A CN201611177306 A CN 201611177306A CN 106718909 B CN106718909 B CN 106718909B
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- dendrobium candidum
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01H—NEW PLANTS OR NON-TRANSGENIC PROCESSES FOR OBTAINING THEM; PLANT REPRODUCTION BY TISSUE CULTURE TECHNIQUES
- A01H4/00—Plant reproduction by tissue culture techniques ; Tissue culture techniques therefor
- A01H4/005—Methods for micropropagation; Vegetative plant propagation using cell or tissue culture techniques
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01H—NEW PLANTS OR NON-TRANSGENIC PROCESSES FOR OBTAINING THEM; PLANT REPRODUCTION BY TISSUE CULTURE TECHNIQUES
- A01H4/00—Plant reproduction by tissue culture techniques ; Tissue culture techniques therefor
- A01H4/008—Methods for regeneration to complete plants
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Abstract
The invention discloses a kind of fast breeding technologies of dendrobium candidum, method includes the following steps: the 1) Fiber differentiation of seed；2) strong sprout and culture of rootage；3) tissue culture transplantation of seedlings.The problems such as plant proliferation obtained the present invention overcomes dendrobium officinale clone culture is slow, quality is low and growing way is inconsistent；Quickly with technical matters simple, breeding easy to operate, free from environmental pollution.Gained dendrobium candidum seedling quality is good, survival rate is high and growing way is uniform etc., provides effective and sustainable method for dendrobium candidum test tube seedling mass propagation and industrialized development.
Description
Technical field
The present invention relates to technical field of pharmaceutical biotechnology, more particularly, to a kind of quick propagating technology of dendrobium candidum.
Background technique
Dendrobium candidum is orchid family (Orchidaceae) Dendrobium (DendrobiumSw) plant, and there are about 500 for whole world orchid family
Belong to, more than 15000 kinds；Dendrobium is one of maximum category in orchid family.There are about 150 categories for Chinese orchid family, and 1000 kinds, stem is cylindrical, high
15~50 cm, thick 4~8 mm.Leaf sheath band meat, square round shape lanceolar, long 3~6.5 cm, wide 0.8~2 cm, hook is omited on top.
Raceme is born in tool leaf or the middle part without base of leaf, spends 2~4；Flower pistac, slightly fragrance；Sepal long 1.2~2
cm；Petal is shorter than sepal, and lip ovum shape lanceolar, long 1.3~1.6 cm wide, 7~9 mm, apex is tapering or short tapering, nearly top
There is round purple patch in centre, has yellow corpus callosum among nearly lower part.4~June of florescence.Surface yellow, base portion slightly gloss, tool
Longitudinal grin has common peduncle trace and remaining leaf sheath on section；Leaf sheath is shorter than internode, often with internode top leaving circular gap, brown, sheath mouth
It opens.Matter is hard and crisp, frangibility, cross-section fiber shape.3~6 mm of fresh goods stem diameter, surface yellow green or blackish green, leaf sheath are greyish white
Color.Gas is micro-, the toughness chewed.It is distributed mainly on provinces and regions on the south the Qinling Mountains and the Yangtze river basin；The most of types of Dendrobium Sw all collect
In be distributed between 15 ° 30 '~25 ° 12 ' of north latitude, and more northwards prolong and type then gradually decreases.Chinese dendrobium candidum mainly divides
It is distributed in the ground such as Zhejiang, Guangxi, Hunan, Yunnan, Guizhou.
It is a kind of rare Chinese medicine, is herbaceos perennial, and stem is grown thickly, cylindrical type, 10--30 centimetres high, slightly
3-8 millimeters, civil, it is known as " giant panda " of " help mesona " medicine circle.Since fresh dendrobium candidum should not save, fresh medicine
It since how fresh and tender its juice is, is easy to rot, especially intermittent drizzles in the rainy season season, it is less easy to maintain, therefore in most of shops of Chinese medicines and doctor
Fresh class medicinal material is rarely provided in Chinese medicine pharmacy of institute, dry product claims Tiepi Fengdou after dendrobium candidum processing, and effective component is mainly dendrobium nobile
Polysaccharide, dendrobine and total amino acid etc..Human immunological competence can be improved, memory, tonifying five zang organs consumptive disease are enhanced, anti-aging inhibits
Tumour, improves diabetic symptom, and anti anoxia has remarkable result to chemicotherapy and night life, excessive drinking and smoking person.
Seeds of Dendrobium Candidum need to could be sprouted without endosperm with certain mycosymbiosises, and natural propagation rate is low, though distribution is wide,
Habitat is narrow, in addition uncontrolled digging all the year round, natural resources is increasingly exhausted, and yield is not able to satisfy existing market demand.
The primary purpose of the present invention is that providing the fast breeding skill of a dendrobium candidum that embryo germination rate is high, survival rate is good
Art.
To achieve the goals above, the invention adopts the following technical scheme:
A kind of fast breeding technical method of dendrobium candidum, comprising the following steps:
1. the Fiber differentiation of seed: removing the withered part of north fruit, after being rinsed well with tap water, impregnate 10- with 70% alcohol
60, then 5-12 min is sterilized with 0.1% mercuric chloride, finally use aseptic water washing 1-5 times.North fruit is aseptically cut, seed
It is seeded in induced medium.Cocounut oil acyl disodium glutamate 1-5g L is added in culture medium- 1, sucrose 10-50gL- 1, agar 5-
15gL- 1, PH4.5-5.5 covers bottle cap, is placed in culturing room and carries out culture
1. strong sprout and culture of rootage:
The dendrobium candidum young shoot of above-mentioned acquisition is taken out from tissue culture bottle, aseptically moves into proliferation and strong seedling culture
In base, bottle cap is covered, is placed in culturing room and is cultivated；
3. tissue culture transplantation of seedlings: dendrobium candidum seedling being transplanted after conventional hardening treatment and is pressed after greenhouse, transplanting
Routine Management, humidity are maintained at 70~90%,
Preferably, 70% alcohol of seed impregnates 20-50s, then sterilizes 6-10 with 0.1% mercuric chloride in above-mentioned steps 1
Min is finally used aseptic water washing 3-4 times.
Preferably, application rate is that seed covers media surface 50%. in above-mentioned steps 1
Preferably, cocounut oil acyl disodium glutamate 1-5g L is added in culture medium in above-mentioned steps 1- 1, sucrose 20-40g
L- 1, agar 8-12gL- 1,Potato extracting solution 100-300mgL- 1,PH4.8-5.2。
Preferably, above-mentioned culture medium is MS culture medium, 1/2MS culture medium, B5 medium, LS culture medium, N6 culture medium.
Preferably, every bottle meets 8 plants of dendrobium candidum young shoots, high 0.3-0.6 cm in above-mentioned steps 2
Preferably, above-mentioned condition of culture is 26-28 DEG C of temperature, intensity of illumination 1600-2000Lx.
Preferably, dendrobium candidum seedling is with 2-5 item root, high 2-3 cm in above-mentioned steps 2.
Preferably, in above-mentioned steps 3, greenhouse is using sawdust, peat soil as matrix, and sawdust: peat soil 1:
1-1:3。
The invention has the following advantages:
1. the present invention will not be to ambient enviroment during Seed inducement culture, strong sprout and culture of rootage, tissue culture transplantation of seedlings
It has an impact, is a kind of environmentally friendly fast breeding technology.
2. what that the present invention overcomes the quality of dendrobium officinale plant strain growth was low, proliferation is slow and growing way is inconsistent etc. asks
Topic, to improve culture efficiency and reduce production cost.The present invention has technical matters is easy to operate, process is few, breeding is quick etc.
Advantage.
Specific embodiment
Below by specific embodiment, technical scheme of the present invention will be further explained in detail.It should be appreciated that this hair
Bright implementation is not limited by the following examples, and the accommodation in any form or change made to the present invention fall within this
Invention protection scope.
Embodiment 1
1. the Fiber differentiation of seed: removing the withered part of north fruit, after being rinsed well with tap water, impregnate 20- with 70% alcohol
50s, then 6-10 min is sterilized with 0.1% mercuric chloride, finally use aseptic water washing 3-4 times.North fruit is aseptically cut, kind
Son is seeded in induced medium, and using MS as minimal medium, cocounut oil acyl disodium glutamate 1-5g L is added in culture medium- 1、
Sucrose 20-40gL- 1, agar 8-12gL- 1,With 1.5mol/LNaOH or 0.2mol/LHCl tune pH4.8-5.2,121
DEG C high-pressure sterilizing pot sterilizes 25 min；Bottle cap is covered, is placed in culturing room and carries out culture
2. strong sprout and culture of rootage:
The dendrobium candidum young shoot of above-mentioned acquisition is taken out from tissue culture bottle, aseptically moves into proliferation and strong seedling culture
In base, bottle cap is covered, is placed in culturing room and is cultivated, 26-28 DEG C of temperature, intensity of illumination 1600-2000Lx.
3. tissue culture transplantation of seedlings: dendrobium candidum seedling is transplanted after conventional hardening treatment in greenhouse, greenhouse
Using sawdust, peat soil as matrix, and sawdust: peat soil 1:1-1:3 is routinely managed after transplanting, and humidity is maintained at 70~
90%。
Embodiment 2
1. the Fiber differentiation of seed: removing the withered part of north fruit, after being rinsed well with tap water, impregnated with 70% alcohol
30s, then 7 min are sterilized with 0.1% mercuric chloride, finally use aseptic water washing 3 times.North fruit is aseptically cut, seed is sowed
In in induced medium, using MS as minimal medium, cocounut oil acyl disodium glutamate 4g L is added in culture medium- 1, sucrose 30g
L- 1, agar 8gL- 1,With 1.5mol/LNaOH or 0.2mol/LHCl tune pH4.8, in 121 DEG C of high-pressure sterilizing pot sterilizings 25
min；Bottle cap is covered, is placed in culturing room and carries out culture
2. strong sprout and culture of rootage:
The dendrobium candidum young shoot of above-mentioned acquisition is taken out from tissue culture bottle, aseptically moves into proliferation and strong seedling culture
In base, bottle cap is covered, is placed in culturing room and is cultivated, 28 DEG C of temperature, intensity of illumination 2000Lx.
3. tissue culture transplantation of seedlings: dendrobium candidum seedling is transplanted after conventional hardening treatment in greenhouse, greenhouse
Using sawdust, peat soil as matrix, and sawdust: peat soil 1:1 is routinely managed after transplanting, and humidity is maintained at 70-90%.
Embodiment 3
1. the Fiber differentiation of seed: removing the withered part of north fruit, after being rinsed well with tap water, impregnated with 70% alcohol
40s, then 5 min are sterilized with 0.1% mercuric chloride, finally use aseptic water washing 4 times.North fruit is aseptically cut, seed is sowed
In in induced medium, using MS as minimal medium, cocounut oil acyl disodium glutamate 3g L is added in culture medium- 1, sucrose 20g
L- 1, agar 12gL- 1,With 1.5mol/LNaOH or 0.2mol/LHCl tune pH5.2, in 121 DEG C of high-pressure sterilizing pot sterilizings 25
min；Cover bottle cap, be placed in culturing room carry out culture oleoyl disodium glutamate and sucrose compatibility can improve dendrobium candidum young shoot
Survival rate.
2. strong sprout and culture of rootage:
The dendrobium candidum young shoot of above-mentioned acquisition is taken out from tissue culture bottle, aseptically moves into proliferation and strong seedling culture
In base, bottle cap is covered, is placed in culturing room and is cultivated, 27 DEG C of temperature, intensity of illumination 1800Lx.
Tissue culture transplantation of seedlings: dendrobium candidum seedling being transplanted after conventional hardening treatment in greenhouse, greenhouse with
Sawdust, peat soil are as matrix, and sawdust: peat soil is routinely to manage after 1:2 transplanting, and humidity is maintained at 70~90%.
Embodiment 4
1. the Fiber differentiation of seed: removing the withered part of north fruit, after being rinsed well with tap water, impregnated with 70% alcohol
30s, then 7 min are sterilized with 0.1% mercuric chloride, finally use aseptic water washing 3 times.North fruit is aseptically cut, seed is sowed
In in induced medium, using MS as minimal medium, cocounut oil acyl disodium glutamate 1g L is added in culture medium- 1, sucrose 30g
L- 1, agar 8gL- 1,With 1.5mol/LNaOH or 0.2mol/LHCl tune pH4.8, in 121 DEG C of high-pressure sterilizing pot sterilizings 25
min；Bottle cap is covered, is placed in culturing room and carries out culture
2. strong sprout and culture of rootage:
The dendrobium candidum young shoot of above-mentioned acquisition is taken out from tissue culture bottle, aseptically moves into proliferation and strong seedling culture
In base, bottle cap is covered, is placed in culturing room and is cultivated, 28 DEG C of temperature, intensity of illumination 2000Lx.
3. tissue culture transplantation of seedlings: dendrobium candidum seedling is transplanted after conventional hardening treatment in greenhouse, greenhouse
Using sawdust, peat soil as matrix, and sawdust: peat soil is routinely to manage after 1:1 transplanting, and humidity is maintained at 70~90%, training
Survival rate, average sprouting number and the average sprouting number greater than 3cm are investigated after supporting 6 months.
| Project | Survival rate | Average new germinative number | Average sprouting number greater than 3cm |
| As a result | 100% | 1.74 | 1.11 |
As seen from the above table, under the program, the dendrobium candidum survival rate of culture 6 months is 100%, and average new germinative number is
1.74, the mean germination number greater than 3 cm is 1.11.
Embodiment 5
1. the Fiber differentiation of seed: removing the withered part of north fruit, after being rinsed well with tap water, impregnated with 70% alcohol
40s, then 5 min are sterilized with 0.1% mercuric chloride, finally use aseptic water washing 4 times.North fruit is aseptically cut, seed is sowed
In in induced medium, using MS as minimal medium, cocounut oil acyl disodium glutamate 1g L is added in culture medium- 1, sucrose 20g
L- 1, agar 12gL- 1,With 1.5mol/LNaOH or 0.2mol/LHCl tune pH5.2, in 121 DEG C of high-pressure sterilizing pot sterilizings 25
min；Bottle cap is covered, is placed in culturing room and carries out culture
2. strong sprout and culture of rootage:
The dendrobium candidum young shoot of above-mentioned acquisition is taken out from tissue culture bottle, aseptically moves into proliferation and strong seedling culture
In base, bottle cap is covered, is placed in culturing room and is cultivated, 27 DEG C of temperature, intensity of illumination 1800Lx.
Tissue culture transplantation of seedlings: dendrobium candidum seedling being transplanted after conventional hardening treatment in greenhouse, greenhouse with
Sawdust, peat soil are as matrix, and sawdust: peat soil is routinely to manage after 1:2 transplanting, and humidity is maintained at 70~90%.Culture
Survival rate, average sprouting number and the average sprouting number greater than 3cm are investigated after 6 months.
| Project | Survival rate | Average new germinative number | Average sprouting number greater than 3cm |
| As a result | 100% | 2.20 | 1.31 |
As seen from the above table, under the program, the dendrobium candidum survival rate of culture 6 months is 100%, and average new germinative number is
2.20, the mean germination number greater than 3 cm is 1.31.
Certainly, above description is not limitation of the present invention, and the present invention is also not limited to the example above.The art
Within the essential scope of the present invention, the variations, modifications, additions or substitutions made also should belong to of the invention those of ordinary skill
Protection scope.
Claims (8)
1. a kind of fast breeding technical method of dendrobium candidum, comprising the following steps:
1) Fiber differentiation of seed:
10-60s after being rinsed well with tap water, is impregnated with 70% alcohol, then disappeared with 0.1% mercuric chloride in the withered part for removing north fruit
Malicious 5-12min is finally used aseptic water washing 1-5 times；North fruit is aseptically cut, seed is seeded in induced medium
On；Cocounut oil acyl disodium glutamate 1-5g L is added in culture medium-1, sucrose 10-50gL-1, agar 5-15gL-1, PH
4.5-5.5 covers bottle cap, is placed in culturing room and is cultivated；
Described cultivate supports base addition cocounut oil acyl disodium glutamate 1-5g L-1, sucrose 20-40gL-1, agar 8-12gL-1,
Potato extracting solution 100-300mgL-1, PH4.8-5.2；
2) strong sprout and culture of rootage:
The dendrobium candidum young shoot of above-mentioned acquisition is taken out from tissue culture bottle, aseptically moves into proliferation and strong seedling culture base
In, bottle cap is covered, is placed in culturing room and is cultivated；
3) tissue culture transplantation of seedlings:
Dendrobium candidum seedling is transplanted after conventional hardening treatment and is routinely managed after greenhouse, transplanting, humidity is kept
70%~90%.
2. a kind of fast breeding technical method of dendrobium candidum according to claim 1, which is characterized in that seed is with 70%
Alcohol impregnates 20-50s, then sterilizes 6-10min with 0.1% mercuric chloride, finally uses aseptic water washing 3-4 times.
3. a kind of fast breeding technical method of dendrobium candidum according to claim 1, which is characterized in that application rate is kind
Son covering media surface 50%.
4. a kind of fast breeding technical method of dendrobium candidum according to claim 1, which is characterized in that above-mentioned culture medium
For MS culture medium, 1/2MS culture medium, B5 medium, LS culture medium, N6 culture medium.
5. a kind of fast breeding technical method of dendrobium candidum according to claim 1, which is characterized in that every bottle connects 8 plants
Dendrobium candidum young shoot, high 0.3-0.6cm.
6. a kind of fast breeding technical method of dendrobium candidum according to claim 1, which is characterized in that above-mentioned culture item
Part is 26-28 DEG C of temperature, intensity of illumination 1600-2000Lx.
7. a kind of fast breeding technical method of dendrobium candidum according to claim 1, which is characterized in that dendrobium candidum children
Seedling is with 2-5 item root, high 2-3cm.
8. a kind of fast breeding technical method of dendrobium candidum according to claim 1, which is characterized in that greenhouse with
Sawdust, peat soil are as matrix, and sawdust: peat soil 1:1-1:3.
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Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101564008A (en) * | 2008-04-24 | 2009-10-28 | 中国医学科学院药用植物研究所 | Hormone-free cultivation and rapid propagation method of dendrobium candidum axenic seedlings |
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2016
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Patent Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101564008A (en) * | 2008-04-24 | 2009-10-28 | 中国医学科学院药用植物研究所 | Hormone-free cultivation and rapid propagation method of dendrobium candidum axenic seedlings |
Non-Patent Citations (1)
| Title |
|---|
| 铁皮石斛组培快繁技术研究;邓瑞云等;《广东药学院学报》;20131231;第29卷(第6期);第609页第2.1至第610页第3.3节 |
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