CN106386499B - A kind of quick breeding method for tissue culture of soil pseudo-ginseng - Google Patents
A kind of quick breeding method for tissue culture of soil pseudo-ginseng Download PDFInfo
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- CN106386499B CN106386499B CN201610927500.9A CN201610927500A CN106386499B CN 106386499 B CN106386499 B CN 106386499B CN 201610927500 A CN201610927500 A CN 201610927500A CN 106386499 B CN106386499 B CN 106386499B
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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 quick breeding method for tissue culture of native pseudo-ginseng；Rhizome including the vigorous tender sprouting of native pseudo-ginseng children of growth selection is explant, liquor potassic permanganate is used successively to explant, Bravo and thiophanate-methyl mixed solution, alcohol and mercuric chloride solution disinfection, culture forms adventitious bud on inducing culture, it is cut again after adventitious bud is cut away partial blade, it is incubated at the proliferation that adventitious bud is carried out in proliferated culture medium, when 3~5 centimetres of subculture height of seedling, it cuts and is inoculated into root media culture, when 5~8 centimetres of height of seedling on root media, in greenhouse natural lighting lower refining seedling 10~15 days, take out test tube seedling, clean root culture medium, Bravo solution impregnates, it is transplanted to by yellow mud, in perlite and the matrix of peat soil mixing, transplanted seedling is obtained after routinely cultivating.The present invention can obtain the test tube seedling that can keep maternal plant merit in a short time, maternal plant be damaged less, and the large-scale production for native pseudo-ginseng provides a large amount of native pseudo-ginseng seedling.
Description
Technical field
The present invention relates to native pseudo-ginsengs, and in particular to a kind of quick breeding method for tissue culture of soil pseudo-ginseng belongs to plant group
Knit culture quick breeding technology field.
Background technology
Native pseudo-ginseng (Stahlianthus involucratus) belongs to Zingiber (Zingiberaceae) Stahlianthes
(Stahlianthus) plant originates in Guangdong, Guangxi, Yunnan, Fujian；India and Sillim are also distributed.Native pseudo-ginseng plant is high about
15~30 centimetres, sepia outside rhizome, inner face brown color, fragrance and have acid, glomeration is expanded in root end；Blade drapes over one's shoulders needle
Shape, blade face bottle green, leaf are longer by green or lilac red, petiole；Inflorescence is extracted out from rhizome, and 10~15 colored consor are in mitriform
Involucre in, involucre green；Grey color, lip center have apricot yellow color spot, interior villosity；It is born in the woods of 800~900 meters of height above sea level
Under, the shady wet place of barren hill；5~July of florescence；Leaf class is seen, it can as hayashishita (Gao Jiangyun waits 2006 for quilt and potting foliage plant；Road
Congress and Wang Yingqiang, 2011).The rhizome tool medical value of native pseudo-ginseng, energy promoting blood circulation to remove blood stasis, swelling and pain relieving cure mainly traumatic injury, wind
Wet ostalgia (《Chinese Plants will》The 2nd fascicle of 16th bundling).
Native pseudo-ginseng can breed, but division propagation speed is slow, and in a manner that cutting rhizome carries out plant division to the root of maternal plant
Stem also has damage.At present, there is not yet carrying out tissue cultures and the report of seedling large-scale production to native pseudo-ginseng, therefore, soil is carried out
The tissue-culturing quick-propagation research of pseudo-ginseng, to improving its sapling multiplication speed, on gardening gardens and as medicinal plant kind
It is of great significance in the popularization and application of plant.
Invention content
Technical problem solved by the invention is to provide a kind of native pseudo-ginseng quick breeding method for tissue culture, can be short-term
The interior test tube seedling for obtaining a large amount of characters and stablizing can keep the good characteristic of maternal plant, the advantage with low input high production.
The purpose of the present invention is achieved through the following technical solutions：
A kind of quick breeding method for tissue culture of soil pseudo-ginseng, includes the following steps：
(a) selection of explant：The rhizome for taking the tender sprouting of eugonic native pseudo-ginseng children is explant；
(b) disinfection of explant：Explant is cleaned with tap water, scissors cuts blade, stays rhizome and high by 5 apart from rhizome
~7 centimetres of stem carries out the disinfection of explant；It cuts away the stem apart from high 1~2 centimetre of rhizome and stays rhizome, and be inoculated with rhizome to luring
It leads in culture medium and cultivates；
(c) induction of Multiple Buds：The culture on inducing culture is until explant forms adventitious bud；The Fiber differentiation
Based component for 1.0~3.0mg/L of MS, 6- benzyl aminoadenine, 0.01~0.10mg/L of methyl α-naphthyl acetate, coconut juice 10.0~15.0%,
20~30g/L of sucrose, 9.5~10g/L of carragheen；
(d) proliferation and subculture：It is cut again after the adventitious bud induced is cut away partial blade, squamous subculture is in proliferated culture medium
The middle proliferation for carrying out adventitious bud；The proliferation and subculture medium component is 3.0~8.0mg/L of MS, 6- benzyl aminoadenine, naphthalene
0.1~0.20mg/L of acetic acid, coconut juice 10.0~15.0%, 20~30g/L of sucrose, 9.5~10g/L of carragheen；
(e) culture of rootage：When 3~5 centimetres of subculture height of seedling, cut and be inoculated into root media, the training of taking root
Support based component for MS, 0.1~1.0mg/L of methyl α-naphthyl acetate, coconut juice 10.0~15.0%, 20~30g/L of sucrose, carragheen 9.5~
10g/L；
(f) acclimatization and transplants：When 5~8 centimetres of height of seedling on root media, in greenhouse natural lighting lower refining seedling 10~15
My god, test tube seedling is taken out, cleans root culture medium, is impregnated 30~40 minutes, is transplanted to 0.1%~0.3% Bravo solution
In matrix by the mixing of yellow mud, perlite and peat soil, keep ventilated, humidity is in 70-85%, and temperature is at 20-32 DEG C, certainly
Transplanted seedling is obtained after right illumination condition culture.
Further to realize the object of the invention, it is preferable that the explant described in step (a) is native pseudo-ginseng
The rhizome of the tender sprouting of children of 5~20 centimetres high of (Stahlianthus involucratus).
Preferably, in terms of mass percent concentration, the disinfection of the explant is first molten with 0.1%-0.2% potassium permanganate
Liquid impregnates rhizome 30~40 minutes, then Bravo and thiophanate-methyl mixed solution with 0.1%~0.3%, impregnates rhizome 40
~50 minutes, then rinse rhizome repeatedly under tap water 30~40 minutes；After drying surface moisture, on superclean bench,
High 4~6 centimetres of stem is cut away, stays 1~2 centimetre high of the stem with rhizome, 70% alcohol wipe rhizome and 1~2 are dipped with cotton balls
Centimetre high stem surface is finally sterilized 15~20 minutes with 0.1% mercuric chloride solution, and aqua sterilisa rinses 4~5 times.
Preferably, the volume ratio of the Bravo and thiophanate-methyl is 1：1.
Preferably, the volume ratio of the yellow mud, perlite and peat soil is 1：1：1.
Preferably, cooled down in step (f) if temperature is higher than 32 DEG C with wind turbine and cascade.
Preferably, the pH of the inducing culture, proliferation and subculture culture medium and root media is 5.6~5.8；Culture
Base sterilising conditions are 125 DEG C, 30 minutes.
Preferably, step (b), step (c), step (d) and step (e) cultivation temperature are 25~30 DEG C, and illumination is strong
It spends for 2000~2300lx, light application time control is 12 hours/day.
Using the explant sterilization method of the present invention, disinfection survival rate can reach 75~85%, 15~25 days can be in explant
The adventitious bud of green is formed on body.
Relative to the prior art, the advantages of the present invention are：
The present invention only needs simple Plant Tissue Breeding equipment that can be completed from explant to test tube in 110~150 days
The a cycle that seedling is formed, greatly accelerates the reproduction speed of native pseudo-ginseng, by shoot proliferation, each shoot proliferation period is not to
Normal bud can be proliferated 4~7 times, therefore, can obtain a large amount of test tube seedling in a short time.The native pseudo-ginseng seedling that the present invention is bred, energy
The good characteristic of maternal plant is kept, transplanting survival rate has the low advantage for putting into high production up to more than 85%.
Description of the drawings
Figure 1A is tender shoots newly long on the native pseudo-ginseng maternal plant of embodiment 2.
Figure 1B is the tender shoots explant removed from native pseudo-ginseng maternal plant of embodiment 2.
Fig. 2 is that the explant mercuric chloride of embodiment 2 sterilizes.
Fig. 3 is the native pseudo-ginseng explant induced synthesis green adventitious bud of embodiment 2.
Fig. 4 is the regeneration plant that the native pseudo-ginseng explant of embodiment 2 obtains.
Fig. 5 A are the seedling of the native pseudo-ginseng test tube seedling transplant survival of embodiment 2.
Fig. 5 B are the seedling of potting training in the secondary transplanting of native pseudo-ginseng of embodiment 2.
Specific embodiment
To more fully understand the present invention, with reference to embodiment, the present invention is further illustrated, but the reality of the present invention
It is unlimited so to apply mode.
Embodiment 1
(a) explant is selected：The rhizome for taking the tender sprouting (5~10 centimetres of height) of eugonic native pseudo-ginseng children is explant；
(b) disinfection of explant：Explant is cleaned with tap water, scissors cuts blade, stays rhizome and apart from rhizome height
About 5~7 centimetres of stem in terms of mass percent concentration, first impregnates rhizome 30 minutes, then use with 0.1% liquor potassic permanganate
0.1% Bravo and thiophanate-methyl (1：1) mixed solution impregnates rhizome 40 minutes, then rinses root repeatedly under tap water
Stem 30 minutes.After drying surface moisture, on superclean bench, high about 4~5 centimetres of stem is cut away, stays about 1~2 centimetre high
Stem with rhizome dips 70% alcohol wipe rhizome and about 1~2 centimetre of high stem surface with cotton balls, finally molten with 0.1% mercuric chloride
Liquid disinfectant 15 minutes, aqua sterilisa rinse 4 times, cut away the stem apart from high about 1~2 centimetre of rhizome and stay rhizome, and are inoculated with rhizome to luring
It leads in culture medium.
(c) induction of Multiple Buds：It is cultivated 3 weeks on inducing culture, there is green bud to be formed on rhizome and grow；Again through 4 weeks,
Sprouting grows 1~2 leaf, about 3~5 centimetres high.The Fiber differentiation based component is MS+6- benzyl aminoadenines 1.0mg/L+
Methyl α-naphthyl acetate 0.01mg/L+10.0% coconut juices+sucrose 30g/L+ carragheens 10g/L.
(d) proliferation and subculture：It is cut again after the adventitious bud induced is cut away partial blade, squamous subculture is in proliferated culture medium
The middle proliferation for carrying out adventitious bud, is cultivated 60 days, can obtain the proliferation Multiple Buds that proliferation times are 4~5.The proliferation and subculture training
It is MS+6- benzyl aminoadenine 3.0mg/L+ methyl α-naphthyl acetates 0.10mg/L+10.0% coconut juices+sucrose 30g/L+ carragheens to support based component
9.5g/L。
(e) culture of rootage：When 3~5 centimetres of subculture height of seedling, cut and be inoculated into root media, culture about 40 days
To rooted seedling.The culture of rootage based component is MS+ methyl α-naphthyl acetates 0.1mg/L+10.0% coconut juices+sucrose 30g/L+ carragheens
9.5g/L。
(f) acclimatization and transplants：When 5~8 centimetres of height of seedling on root media, in greenhouse natural lighting lower refining seedling 10 days,
Test tube seedling is taken out, cleans root culture medium, 0.1% Bravo solution is impregnated 30 minutes, is transplanted to by yellow mud, perlite and mud
Charcoal soil (1：1：1) in the matrix mixed in equal volume, keep ventilated, humidity is in 70-85%, and temperature is at 20-32 DEG C, higher than 32
DEG C wind turbine and cascade need to be used to cool down, transplanted seedling is obtained after natural lighting CMC model.
In above-mentioned tetra- steps of b, c, d, e, culture medium used：Inducing culture, proliferation and subculture culture medium and training of taking root
The pH for supporting base is 5.6~5.8, and medium sterilization condition is 125 DEG C, and 30 minutes, condition of culture was 25~30 DEG C, intensity of illumination
2000~2300lx, 12 hour/day of illumination.
Transplanted seedling pays attention to divulging information, water and shading, and for survival rate up to more than 90%, transplanting can go up potting training after 45 days.From
Selection explant about needs the time of 149 days to regeneration test tube seedling.
Embodiment 2
(a) explant is selected：As shown in Figure 1A, it is the tender sprouting of children grown on native pseudo-ginseng maternal plant, takes 8~15 centimetres high
The rhizome of the tender sprouting of children is explant.
(b) disinfection of explant：As shown in Figure 1B, it is the tender sprouting explant of children under being cut from native pseudo-ginseng maternal plant, first
It is cleaned with tap water, scissors cuts blade, rhizome and the stem apart from high about 5~7 centimetres of rhizome is stayed, first with 0.2% potassium permanganate
Solution impregnates rhizome 30 minutes, then Bravo and thiophanate-methyl (1 with 0.2%：1) mixed solution impregnates rhizome 40 minutes,
Then rhizome is rinsed repeatedly under tap water 40 minutes.After drying surface moisture, on superclean bench, cut away about 4~6 lis high
The stem of rice, stays about 1~2 centimetre of high stem with rhizome, and 70% alcohol wipe rhizome and about 1~2 centimetre high are dipped with cotton balls
Stem surface, last as shown in Fig. 2, being sterilized 18 minutes with 0.1% mercuric chloride solution, aqua sterilisa rinses 4 times, cuts away high about apart from rhizome
1~2 centimetre of stem stays rhizome, and is inoculated in rhizome to inducing culture.
(c) induction of Multiple Buds：It is cultivated 2 weeks on inducing culture, as shown in figure 3, thering is green bud to be formed on rhizome and growing
Go out；Again through 4 weeks, sprouting grows 1~2 leaf, about 3~5 centimetres high.The Fiber differentiation based component is MS+6- benzyl amino glands
Purine 2.0mg/L+ methyl α-naphthyl acetates 0.05mg/L+15.0% coconut juices+sucrose 20g/L+ carragheens 9.5g/L.
(d) proliferation and subculture：It is cut again after the adventitious bud induced is cut away partial blade, squamous subculture is in proliferated culture medium
The middle proliferation for carrying out adventitious bud, is cultivated 50 days, can obtain the proliferation Multiple Buds that proliferation times are 4~6.Proliferation and subculture culture medium into
It is divided into MS+6- benzyl aminoadenine 5.0mg/L+ methyl α-naphthyl acetates 0.20mg/L+15.0% coconut juices+sucrose 20g/L+ carragheens 9.5g/
L。
(e) culture of rootage：It when 3~5 centimetres of subculture height of seedling, cuts and is inoculated into root media, after culture 35 days, such as
Shown in Fig. 4, rooted seedling is obtained.The culture of rootage based component is MS+ methyl α-naphthyl acetates 0.5mg/L+15.0% coconut juices+sucrose 20g/
L+ carragheens 9.5g/L.
(f) acclimatization and transplants：When 5~8 centimetres of height of seedling on root media, in greenhouse natural lighting lower refining seedling 12 days,
Take out test tube seedling, clean root culture medium, 0.2% Bravo solution impregnates 30 minutes, then as shown in Figure 5A, be transplanted to by
Yellow mud, perlite and peat soil (1：1：1) in the matrix mixed in equal volume, keep ventilated, humidity is in 70-85%, temperature
At 20-32 DEG C, wind turbine and cascade need to be used to cool down higher than 32 DEG C, transplanted seedling is obtained after natural lighting CMC model.
In above-mentioned (b), (c), (d), (e) four steps, culture medium used：Inducing culture, proliferation and subculture culture medium
PH with root media is 5.6~5.8, and medium sterilization condition is 125 DEG C, and 30 minutes, condition of culture was 25~30 DEG C, light
According to 2000~2300lx of intensity, 12 hour/day of illumination.
Transplanted seedling pays attention to divulging information, water and shading, and survival rate is such as schemed up to more than 85%, upper potting training after transplanting 45 days
Shown in 5B, the seedling for potting training in the secondary transplanting of native pseudo-ginseng seedling.127 days are about needed to regeneration test tube seedling from selection explant
Time.
Embodiment 3
(a) explant is selected：The rhizome for taking the tender sprouting (10~20 centimetres of height) of eugonic native pseudo-ginseng children is explant；
(b) disinfection of explant：Explant is cleaned with tap water, scissors cuts blade, stays rhizome and apart from rhizome height
About 5~7 centimetres of stem first impregnates rhizome 40 minutes, then Bravo and methyl sulphur with 0.3% with 0.1% liquor potassic permanganate
Bacterium spirit (1：1) mixed solution impregnates rhizome 50 minutes, then rinses rhizome repeatedly under tap water 40 minutes.Dry surface moisture
Afterwards, on superclean bench, high about 4~6 centimetres of stem is cut away, about 1~2 centimetre of high stem with rhizome is stayed, is dipped with cotton balls
70% alcohol wipe rhizome and about 1~2 centimetre of high stem surface are finally sterilized 20 minutes with 0.1% mercuric chloride solution, aqua sterilisa punching
It washes 5 times, cuts away the stem apart from high about 1~2 centimetre of rhizome and stay rhizome, and be inoculated in rhizome to inducing culture.
(c) induction of Multiple Buds：It is cultivated 2 weeks on inducing culture, there is green bud to be formed on rhizome and grow；Again through 3 weeks,
Sprouting grows 1~2 leaf, about 3~5 centimetres high.The Fiber differentiation based component is MS+6- benzyl aminoadenines 3.0mg/L+
Methyl α-naphthyl acetate 0.10mg/L+15.0% coconut juices+sucrose 30g/L+ carragheens 10g/L.
(d) proliferation and subculture：It is cut again after the adventitious bud induced is cut away partial blade, squamous subculture is in proliferated culture medium
The middle proliferation for carrying out adventitious bud, is cultivated 45 days, can obtain the proliferation Multiple Buds that proliferation times are 5~7.The proliferation and subculture training
It is MS+6- benzyl aminoadenine 8.0mg/L+ methyl α-naphthyl acetates 0.20mg/L+15.0% coconut juices+sucrose 30g/L+ carragheens to support based component
10g/L。
(e) culture of rootage：It when 3~5 centimetres of subculture height of seedling, cuts and is inoculated into root media, culture obtains for 30 days
Rooted seedling.The culture of rootage based component is MS+ methyl α-naphthyl acetates 1.0mg/L+15.0% coconut juices+sucrose 30g/L+ carragheens
9.5g/L。
(f) acclimatization and transplants：When 5~8 centimetres of height of seedling on root media, in greenhouse natural lighting lower refining seedling 15 days,
Test tube seedling is taken out, cleans root culture medium, 0.3% Bravo solution is impregnated 40 minutes, is transplanted to by yellow mud, perlite and peat
Soil (1：1：1) in the matrix mixed in equal volume, keep ventilated, humidity is in 70-85%, and temperature is at 20-32 DEG C, higher than 32 DEG C
Wind turbine and cascade need to be used to cool down, transplanted seedling is obtained after natural lighting CMC model.
In above-mentioned tetra- steps of b, c, d, e, culture medium used：Inducing culture, proliferation and subculture culture medium and training of taking root
The pH for supporting base is 5.6~5.8, and medium sterilization condition is 125 DEG C, and 30 minutes, condition of culture was 25~30 DEG C, intensity of illumination
2000~2300lx, 12 hour/day of illumination.
Transplanted seedling pays attention to divulging information, water and shading, and for survival rate up to more than 85%, transplanting can go up potting training after 45 days.From
Selection explant about needs the time of 110 days to regeneration test tube seedling.
Comparative example 1
The explant disinfection way of this comparative example soil pseudo-ginseng is：Explant is cleaned with tap water, scissors cuts blade, stays
Rhizome and the stem apart from high about 5~7 centimetres of rhizome, under tap water plus a small amount of washing powder rinses rhizome 40 minutes repeatedly.It dries
After surface moisture, on superclean bench, high about 4~6 centimetres of stem is cut away, stays about 1~2 centimetre of high stem with rhizome, is used
Cotton balls dips 70% alcohol wipe rhizome and about 1~2 centimetre of high stem surface, is finally sterilized 10 minutes with 0.1% mercuric chloride solution,
Aqua sterilisa rinses 4 times, cuts away the stem apart from high about 1~2 centimetre of rhizome and stays rhizome, and be inoculated in rhizome to inducing culture.This
The inducing culture of comparative example and the condition of embodiment 3 are all identical.Inoculation 1 week or so is it has been observed that inducing culture 100% is dirty
Dye.
Comparative example 2
The explant disinfection way of this comparative example soil pseudo-ginseng is：Explant is cleaned with tap water, scissors cuts blade, stays
Rhizome and the stem apart from high about 5~7 centimetres of rhizome, rinse rhizome 40 minutes repeatedly under tap water.After drying surface moisture,
On superclean bench, high about 4~6 centimetres of stem is cut away, about 1~2 centimetre of high stem with rhizome is stayed, 70% wine is dipped with cotton balls
Essence wiping rhizome and about 1~2 centimetre of high stem surface, are finally sterilized 15 minutes with 0.1% mercuric chloride solution, and aqua sterilisa rinses 5 times,
It cuts away the stem apart from high about 1~2 centimetre of rhizome and stays rhizome, and be inoculated in rhizome to inducing culture.The induction training of this comparative example
The condition for supporting base and embodiment 3 is all identical.Inoculation 1 week or so is it has been observed that inducing culture 100% pollutes.
The advantages of embodiment of the present invention is relative to comparative example is as follows：Due to the difference of explant sterilization method, the present invention is real
It applies example and greatly reduces pollution rate of the explant in inducing culture, so as to improve work efficiency.
Claims (7)
1. a kind of quick breeding method for tissue culture of soil pseudo-ginseng, it is characterised in that include the following steps：
(a) selection of explant：The rhizome for taking the tender sprouting of eugonic native pseudo-ginseng children is explant；
(b) disinfection of explant：Explant is cleaned with tap water, scissors cuts blade, stays rhizome and high by 5~7 apart from rhizome
Centimetre stem, carry out the disinfection of explant；It cuts away the stem apart from high 1~2 centimetre of rhizome and stays rhizome, and be inoculated with rhizome and trained to induction
It supports and is cultivated in base；In terms of mass percent concentration, the disinfection of the explant is first to be soaked with 0.1%-0.2% liquor potassic permanganates
It steeps rhizome 30~40 minutes, then Bravo and thiophanate-methyl mixed solution with 0.1%~0.3%, impregnates rhizome 40~50
Minute, then rinse rhizome repeatedly under tap water 30~40 minutes；After drying surface moisture, on superclean bench, cut away
High 4~6 centimetres of stem stays 1~2 centimetre high of the stem with rhizome, and 70% alcohol wipe rhizome and 1~2 centimetre are dipped with cotton balls
High stem surface is finally sterilized 15~20 minutes with 0.1% mercuric chloride solution, and aqua sterilisa rinses 4~5 times；
(c) induction of Multiple Buds：The culture on inducing culture is until explant forms adventitious bud；The inducing culture into
It is divided into 1.0~3.0mg/L of MS, 6- benzyl aminoadenine, 0.01~0.10mg/L of methyl α-naphthyl acetate, coconut juice 10.0~15.0%, sucrose
20~30g/L, 9.5~10g/L of carragheen；
(d) proliferation and subculture：Cut again after the adventitious bud induced is cut away partial blade, squamous subculture in proliferated culture medium into
The proliferation of row adventitious bud；The proliferation and subculture medium component is 3.0~8.0mg/L of MS, 6- benzyl aminoadenine, methyl α-naphthyl acetate
0.1~0.20mg/L, coconut juice 10.0~15.0%, 20~30g/L of sucrose, 9.5~10g/L of carragheen；
(e) culture of rootage：When 3~5 centimetres of subculture height of seedling, cut and be inoculated into root media, the root media
Ingredient is MS, 0.1~1.0mg/L of methyl α-naphthyl acetate, coconut juice 10.0~15.0%, 20~30g/L of sucrose, 9.5~10g/L of carragheen；
(f) acclimatization and transplants：When 5~8 centimetres of height of seedling on root media, in greenhouse natural lighting lower refining seedling 10~15 days,
Test tube seedling is taken out, cleans root culture medium, is impregnated 30~40 minutes, is transplanted to by Huang with 0.1%~0.3% Bravo solution
In the matrix of mud, perlite and peat soil mixing, keep ventilated, humidity is in 70-85%, and temperature is at 20-32 DEG C, natural light
According to obtaining transplanted seedling after CMC model.
2. the quick breeding method for tissue culture of soil pseudo-ginseng according to claim 1, it is characterised in that：Step (a) is described
Explant be native pseudo-ginseng (Stahlianthus involucratus) 5~20 centimetres high of the tender sprouting of children rhizome.
3. the quick breeding method for tissue culture of soil pseudo-ginseng according to claim 1, it is characterised in that：The Bravo and
The volume ratio of thiophanate-methyl is 1：1.
4. the quick breeding method for tissue culture of soil pseudo-ginseng according to claim 1, it is characterised in that：The yellow mud, treasure
The volume ratio of Zhu Yan and peat soil is 1：1：1.
5. the quick breeding method for tissue culture of soil pseudo-ginseng according to claim 1, it is characterised in that：In step (5) such as
Fruit temperature is higher than 32 DEG C, is cooled down with wind turbine and cascade.
6. the quick breeding method for tissue culture of soil pseudo-ginseng according to claim 1, it is characterised in that：The Fiber differentiation
The pH of base, proliferation and subculture culture medium and root media is 5.6~5.8；Medium sterilization condition is 125 DEG C, 30 minutes.
7. the quick breeding method for tissue culture of soil pseudo-ginseng according to claim 1, it is characterised in that：Step (b), step
(c), the temperature of step (d) and step (e) culture is 25~30 DEG C, and intensity of illumination is 2000~2300lx, light application time
It controls as 12 hours/day.
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| CN107182783A (en) * | 2017-06-13 | 2017-09-22 | 广东省农业科学院环境园艺研究所 | A kind of method of Hainan rhizomes of Panax notoginseng bastem portion regeneration induction plant |
| CN107593443B (en) * | 2017-10-18 | 2019-12-20 | 广东省农业科学院环境园艺研究所 | Tissue culture rapid propagation method of kaempferia galanga |
| CN108293824A (en) * | 2017-12-26 | 2018-07-20 | 广东省农业科学院环境园艺研究所 | The method that a kind of potting Hainan Radix Notoginseng is downgraded |
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