CN106386499A - Tissue cultured rapid propagation method for Rhizoma Stahlianthi Involucrati - Google Patents
Tissue cultured rapid propagation method for Rhizoma Stahlianthi Involucrati Download PDFInfo
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- CN106386499A CN106386499A CN201610927500.9A CN201610927500A CN106386499A CN 106386499 A CN106386499 A CN 106386499A CN 201610927500 A CN201610927500 A CN 201610927500A CN 106386499 A CN106386499 A CN 106386499A
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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 tissue cultured rapid propagation method for Rhizoma Stahlianthi Involucrati. The method comprises the following steps: selecting a rhizome of a young and tender bud of Rhizoma Stahlianthi Involucrati which grows vigorously as an explant; sterilizing the explant by successively using a potassium permanganate solution, a chlorothalonil and thiophanate-methyl mixed solution, alcohol and a mercury chloride solution; culturing on an induction medium to form an adventitious bud; after cutting off part of lamina of the adventitious bud, cutting the adventitious bud; culturing in a proliferation medium and proliferating the adventitious bud; when the height of a subculture seedling is 3-5 centimeters, cutting off the subculture seedling and inoculating into a rooting medium for culturing; when the height of a seedling on the rooting medium is 5-8 centimeters, hardening the seedling under natural illumination of a greenhouse; taking out a test-tube seedling; cleaning the medium of the root of the test-tube seedling; soaking with a chlorothalonil solution; transplanting into a matrix obtained by mixing yellow mud, perlite and peat soil; and culturing conventionally to obtain a transplanting seedling. The test-tube seedling which has excellent characters of a mother plant can be obtained within a short time, injury to the mother plant is little, and a large number of Rhizoma Stahlianthi Involucrati seedlings are provided for large-scale production of Rhizoma Stahlianthi Involucrati.
Description
Technical field
The present invention relates to Rhizoma Stahlianthi Involucrati, and in particular to a kind of quick breeding method for tissue culture of Rhizoma Stahlianthi Involucrati, belongs to plant group
Knit culture quick breeding technology field.
Background technology
Rhizoma Stahlianthi Involucrati (Stahlianthus involucratus) belongs to Zingiberaceae (Zingiberaceae) Stahlianthes
(Stahlianthus) plant, originates in Guangdong, Guangxi, Yunnan, Fujian；India and Sillim are also distributed.Rhizoma Stahlianthi Involucrati 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 pin
Shape, blade face bottle green, by green or lilac red, petiole is longer for leaf；Inflorescence is extracted out from rhizome, and 10～15 colored consor are in mitriform
Involucre in, involucre green；Floral white, lip central authorities have apricot yellow mottle, 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；See leaf class, can as sylvan life quilt and potted plant foliage plant (Gao Jiangyun waits 2006；Road
Congress and Wang Yingqiang, 2011).The rhizome tool medical value of Rhizoma Stahlianthi Involucrati, energy promoting blood circulation to remove blood stasis, reducing swelling and alleviating pain, cure mainly traumatic injury, wind
Wet osteodynia (《Chinese Plants will》16th bundling the 2nd fascicle).
Rhizoma Stahlianthi Involucrati can carry out breeding by way of plant division by cutting rhizome, but division propagation speed is slow, and the root to maternal plant
Stem also has damage.At present, there is not yet Rhizoma Stahlianthi Involucrati is carried out with tissue culture and the report of seedling large-scale production, therefore, carry out soil
The tissue-culturing quick-propagation research of Radix Notoginseng, to its sapling multiplication speed of raising, on gardening gardens with as medicinal plants kind
Significant in the popularization and application planted.
Content of the invention
Technical problem solved by the invention is to provide a kind of Rhizoma Stahlianthi Involucrati quick breeding method for tissue culture, can be in short-term
The stable test tube seedling of a large amount of character of interior acquisition, can keep the good characteristic of maternal plant, have the low advantage putting into 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 Rhizoma Stahlianthi Involucrati, comprises the following steps：
The selection of (a) explant：The rhizome taking the tender sprouting of eugonic Rhizoma Stahlianthi Involucrati children is explant；
The sterilization of (b) explant：Explant tap water is cleaned, blade cut by shears, stay rhizome and high by 5 apart from rhizome
～7 centimetres of stem, carries out the sterilization of explant；Cut away the stem apart from high 1～2 centimetre of rhizome and stay rhizome, and inoculate rhizome to luring
Lead culture in culture medium；
The induction of (c) Multiple Buds：Culture on inducing culture is until explant forms adventitious bud；Described inducing culture
Based component be MS, 6 benzyl aminoadenine 1.0～3.0mg/L, naphthalene acetic acid 0.01～0.10mg/L, coconut juice 10.0～15.0%,
Sucrose 20～30g/L, carrageenan 9.5～10g/L；
(d) proliferation and subculture：The adventitious bud inducing is cut away and cuts again after partial blade, successive transfer culture is in proliferated culture medium
In carry out the propagation of adventitious bud；Described proliferation and subculture medium component is MS, 6 benzyl aminoadenine 3.0～8.0mg/L, naphthalene
Acetic acid 0.1～0.20mg/L, coconut juice 10.0～15.0%, sucrose 20～30g/L, carrageenan 9.5～10g/L；
(e) root culture：When 3～5 centimetres of subculture height of seedling, cut and be inoculated in root media, described training of taking root
Foster based component is MS, naphthalene acetic acid 0.1～1.0mg/L, coconut juice 10.0～15.0%, sucrose 20～30g/L, carrageenan 9.5～
10g/L；
(f) acclimatization and transplantses：When 5～8 centimetres of the height of seedling on root media, in greenhouse natural lighting lower refining seedling 10～15
My god, take out test tube seedling, clean root culture medium, the Bravo solution soaking with 0.1%～0.3% 30～40 minutes, be transplanted to
In substrate by the mixing of yellow mud, perlite and peat soil, keep ventilated, humidity 70 85%, temperature at 20 32 DEG C, from
So obtain transplanted seedling after illumination condition culture.
For realizing the object of the invention further it is preferable that the explant described in step (a) is Rhizoma Stahlianthi Involucrati
The rhizome of the tender sprouting of 5～20 centimetres of high childrens of (Stahlianthus involucratus).
Preferably, in terms of mass percent concentration, the sterilization of described explant is first molten with 0.1% 0.2% potassium permanganate
Immersion bubble rhizome 30～40 minutes, then the Bravo with 0.1%～0.3% and thiophanate-methyl mixed solution, soak rhizome 40
～50 minutes, then repeatedly rinse rhizome under tap water 30～40 minutes；After drying surface moisture, on superclean bench,
Cut away high 4～6 centimetres of stem, stay 1～2 centimetre high of the stem with rhizome, dip 70% alcohol wipe rhizome and 1～2 with cotton balls
Centimetre high stem surface, is finally sterilized 15～20 minutes with 0.1% mercuric chloride solution, aquesterilisa rinses 4～5 times.
Preferably, the volume ratio of described Bravo and thiophanate-methyl is 1：1.
Preferably, the volume ratio of described yellow mud, perlite and peat soil is 1：1：1.
Preferably, if temperature is higher than 32 DEG C in step (f), with blower fan and cascade cooling.
Preferably, the pH of described 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 the described cultivation temperature of step (e) are 25～30 DEG C, and illumination is strong
Spend for 2000～2300lx, light application time is controlled to 12 hours/day.
The explant sterilization method of the application present invention, sterilization survival rate can reach 75～85%, 15～25 days can be in explant
The adventitious bud of green is formed on body.
With respect to prior art, advantages of the present invention and having the beneficial effect that：
The present invention only needs simple plant tissue culture equipment can complete from explant to test tube in 110～150 days
The a cycle that Seedling is formed, greatly accelerates the reproduction speed of Rhizoma Stahlianthi Involucrati, through subculture multiplication, each subculture multiplication cycle is not to
Normal bud can breed 4～7 times, therefore, can obtain substantial amounts of test tube seedling at short notice.The Rhizoma Stahlianthi Involucrati seedling of present invention breeding, energy
Keep the good characteristic of maternal plant, transplanting survival rate, up to more than 85%, has the low advantage putting into high production.
Brief description
Figure 1A is the tender shoots newly growing on the Rhizoma Stahlianthi Involucrati maternal plant of embodiment 2.
Figure 1B is the tender shoots explant taking off from Rhizoma Stahlianthi Involucrati maternal plant of embodiment 2.
Fig. 2 is the explant mercuric chloride sterilization of embodiment 2.
Fig. 3 is the Rhizoma Stahlianthi Involucrati explant induced synthesis green adventitious bud of embodiment 2.
Fig. 4 is the regeneration plant of the Rhizoma Stahlianthi Involucrati explant acquisition of embodiment 2.
Fig. 5 A is the seedling of the Rhizoma Stahlianthi Involucrati test tube seedling transplant survival of embodiment 2.
Fig. 5 B is the secondary seedling transplanting upper potted plant training of Rhizoma Stahlianthi Involucrati of embodiment 2.
Specific embodiment
For more fully understanding the present invention, with reference to embodiment, the present invention is further illustrated, but the reality of the present invention
Mode of applying does not limit so.
Embodiment 1
A () selects explant：The rhizome taking the tender sprouting (5～10 centimetres high) of eugonic Rhizoma Stahlianthi Involucrati children is explant；
The sterilization of (b) explant：Explant tap water is cleaned, blade cut by shears, stay rhizome and apart from rhizome height
About 5～7 centimetres of stem, in terms of mass percent concentration, first soaks rhizome 30 minutes with 0.1% potassium permanganate solution, then uses
0.1% Bravo and thiophanate-methyl (1：1) mixed solution soaks rhizome 40 minutes, then repeatedly rinses root under tap water
Stem 30 minutes.After drying surface moisture, on superclean bench, cut away high about 4～5 centimetres of stem, stay about 1～2 centimetre high
With the stem of rhizome, dip 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, aquesterilisa rinses 4 times, cuts away the stem apart from high about 1～2 centimetre of rhizome and stays rhizome, and inoculates rhizome to luring
Lead in culture medium.
The induction of (c) Multiple Buds：Inducing culture is cultivated 3 weeks, rhizome has green bud formed and grow；Again through 4 weeks,
Sprouting grows 1～2 leaf, high about 3～5 centimetres.Described inducing culture based component is MS+6 benzyl aminoadenine 1.0mg/L+
Naphthalene acetic acid 0.01mg/L+10.0% coconut juice+sucrose 30g/L+ carrageenan 10g/L.
(d) proliferation and subculture：The adventitious bud inducing is cut away and cuts again after partial blade, successive transfer culture is in proliferated culture medium
In carry out the propagation of adventitious bud, cultivate 60 days, the propagation Multiple Buds that proliferation times are 4～5 can be obtained.Described proliferation and subculture training
Foster based component is MS+6 benzyl aminoadenine 3.0mg/L+ naphthalene acetic acid 0.10mg/L+10.0% coconut juice+sucrose 30g/L+ carrageenan
9.5g/L.
(e) root culture：When 3～5 centimetres of subculture height of seedling, cut and be inoculated in root media, culture about 40 days
To Seedling of taking root.Described root culture based component is MS+ naphthalene acetic acid 0.1mg/L+10.0% coconut juice+sucrose 30g/L+ carrageenan
9.5g/L.
(f) acclimatization and transplantses：When 5～8 centimetres of the height of seedling on root media, in greenhouse natural lighting lower refining seedling 10 days,
Take out test tube seedling, clean root culture medium, 0.1% Bravo solution soaking 30 minutes, be transplanted to by yellow mud, perlite and mud
Charcoal soil (1：1：1), in the substrate of equal-volume mixing, keep ventilated, humidity 70 85%, temperature at 20 32 DEG C, higher than 32
DEG C need to blower fan and cascade cooling, obtain transplanted seedling 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 of foster base is 5.6～5.8, and medium sterilization condition is 125 DEG C, 30 minutes, and condition of culture is 25～30 DEG C, intensity of illumination
2000～2300lx, illumination 12 hour/day.
Transplanted seedling notes ventilation, waters and shade, and survival rate, up to more than 90%, can go up potted plant training after transplanting 45 days.From
Explant is selected about to need the time of 149 days to regeneration test tube seedling.
Embodiment 2
A () selects explant：As shown in Figure 1A, it is the tender sprouting of the children growing on Rhizoma Stahlianthi Involucrati maternal plant, take 8～15 centimetres high
The rhizome of the tender sprouting of children is explant.
The sterilization of (b) explant：As shown in Figure 1B, it is children's tender sprouting explant under cutting from Rhizoma Stahlianthi Involucrati maternal plant, first
Cleaned with tap water, blade cut by shears, stay rhizome and the stem apart from high about 5～7 centimetres of rhizome, first use 0.2% potassium permanganate
Solution soaking rhizome 30 minutes, then the Bravo with 0.2% and thiophanate-methyl (1：1) mixed solution soaks rhizome 40 minutes,
Then repeatedly rinse rhizome under tap water 40 minutes.After drying surface moisture, on superclean bench, cut away high about 4～6 lis
The stem of rice, stays about 1～2 centimetre of high stem with rhizome, dips 70% alcohol wipe rhizome and about 1～2 centimetre high with cotton balls
Stem surface, finally as shown in Fig. 2 being sterilized 18 minutes with 0.1% mercuric chloride solution, aquesterilisa rinses 4 times, cuts away high about apart from rhizome
1～2 centimetre of stem stays rhizome, and inoculates rhizome in inducing culture.
The induction of (c) Multiple Buds：Inducing culture is cultivated 2 weeks, as shown in figure 3, there being green bud to be formed and grow on rhizome
Go out；Again through 4 weeks, sprouting grows 1～2 leaf, high about 3～5 centimetres.Described inducing culture based component is MS+6 benzyl amino gland
Purine 2.0mg/L+ naphthalene acetic acid 0.05mg/L+15.0% coconut juice+sucrose 20g/L+ carrageenan 9.5g/L.
(d) proliferation and subculture：The adventitious bud inducing is cut away and cuts again after partial blade, successive transfer culture is in proliferated culture medium
In carry out the propagation of adventitious bud, cultivate 50 days, the propagation Multiple Buds that proliferation times are 4～6 can be obtained.Proliferation and subculture culture medium becomes
It is divided into MS+6 benzyl aminoadenine 5.0mg/L+ naphthalene acetic acid 0.20mg/L+15.0% coconut juice+sucrose 20g/L+ carrageenan 9.5g/
L.
(e) root culture：When 3～5 centimetres of subculture height of seedling, cut and be inoculated in root media, after culture 35 days, such as
Shown in Fig. 4, obtain Seedling of taking root.Described root culture based component is MS+ naphthalene acetic acid 0.5mg/L+15.0% coconut juice+sucrose 20g/
L+ carrageenan 9.5g/L.
(f) acclimatization and transplantses：When 5～8 centimetres of the 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 soaking 30 minutes, then as shown in Figure 5A, be transplanted to by
Yellow mud, perlite and peat soil (1：1：1), in the substrate of equal-volume mixing, keep ventilated, humidity 70 85%, temperature
At 20 32 DEG C, transplanted seedling need to be obtained with blower fan and cascade cooling after natural lighting CMC model higher than 32 DEG C.
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, 30 minutes, and condition of culture is 25～30 DEG C, light
According to intensity 2000～2300lx, illumination 12 hour/day.
Transplanted seedling notes ventilation, waters and shade, and up to more than 85%, after transplanting 45 days, upper potted plant training, such as schemes survival rate
Shown in 5B, it is the secondary seedling transplanting upper potted plant training of Rhizoma Stahlianthi Involucrati seedling.About need 127 days from selecting explant to regeneration test tube seedling
Time.
Embodiment 3
A () selects explant：The rhizome taking the tender sprouting (10～20 centimetres high) of eugonic Rhizoma Stahlianthi Involucrati children is explant；
The sterilization of (b) explant：Explant tap water is cleaned, blade cut by shears, stay rhizome and apart from rhizome height
About 5～7 centimetres of stem, first soaks rhizome 40 minutes with 0.1% potassium permanganate solution, then the Bravo with 0.3% and methyl sulfur
Bacterium spirit (1：1) mixed solution soaks rhizome 50 minutes, then repeatedly rinses rhizome under tap water 40 minutes.Dry surface moisture
Afterwards, on superclean bench, cut away high about 4～6 centimetres of stem, stay about 1～2 centimetre of high stem with rhizome, 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, aquesterilisa rushes
Wash 5 times, cut away the stem apart from high about 1～2 centimetre of rhizome and stay rhizome, and inoculate rhizome in inducing culture.
The induction of (c) Multiple Buds：Inducing culture is cultivated 2 weeks, rhizome has green bud formed and grow；Again through 3 weeks,
Sprouting grows 1～2 leaf, high about 3～5 centimetres.Described inducing culture based component is MS+6 benzyl aminoadenine 3.0mg/L+
Naphthalene acetic acid 0.10mg/L+15.0% coconut juice+sucrose 30g/L+ carrageenan 10g/L.
(d) proliferation and subculture：The adventitious bud inducing is cut away and cuts again after partial blade, successive transfer culture is in proliferated culture medium
In carry out the propagation of adventitious bud, cultivate 45 days, the propagation Multiple Buds that proliferation times are 5～7 can be obtained.Described proliferation and subculture training
Foster based component is MS+6 benzyl aminoadenine 8.0mg/L+ naphthalene acetic acid 0.20mg/L+15.0% coconut juice+sucrose 30g/L+ carrageenan
10g/L.
(e) root culture：When 3～5 centimetres of subculture height of seedling, cut and be inoculated in root media, culture obtains for 30 days
Take root Seedling.Described root culture based component is MS+ naphthalene acetic acid 1.0mg/L+15.0% coconut juice+sucrose 30g/L+ carrageenan
9.5g/L.
(f) acclimatization and transplantses：When 5～8 centimetres of the height of seedling on root media, in greenhouse natural lighting lower refining seedling 15 days,
Take out test tube seedling, clean root culture medium, 0.3% Bravo solution soaking 40 minutes, be transplanted to by yellow mud, perlite and peat
Soil (1：1：1), in the substrate of equal-volume mixing, keep ventilated, humidity 70 85%, temperature at 20 32 DEG C, higher than 32 DEG C
Transplanted seedling need to be obtained with blower fan and cascade cooling 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 of foster base is 5.6～5.8, and medium sterilization condition is 125 DEG C, 30 minutes, and condition of culture is 25～30 DEG C, intensity of illumination
2000～2300lx, illumination 12 hour/day.
Transplanted seedling notes ventilation, waters and shade, and survival rate, up to more than 85%, can go up potted plant training after transplanting 45 days.From
Explant is selected about to need the time of 110 days to regeneration test tube seedling.
Comparative example 1
The explant disinfection way of this comparative example Rhizoma Stahlianthi Involucrati is：Explant tap water is cleaned, blade cut by shears, stay
Rhizome and the stem apart from high about 5～7 centimetres of rhizome, under tap water plus a small amount of detergent rinses rhizome 40 minutes repeatedly.Dry
After surface moisture, on superclean bench, cut away high about 4～6 centimetres of stem, stay about 1～2 centimetre of high stem with rhizome, use
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,
Aquesterilisa rinses 4 times, cuts away the stem apart from high about 1～2 centimetre of rhizome and stays rhizome, and inoculates rhizome in inducing culture.This
The inducing culture of comparative example is all identical with the condition of embodiment 3.Inoculation 1 week about is it has been observed that inducing culture 100% is dirty
Dye.
Comparative example 2
The explant disinfection way of this comparative example Rhizoma Stahlianthi Involucrati is：Explant tap water is cleaned, blade cut by shears, stay
Rhizome and the stem apart from high about 5～7 centimetres of rhizome, rinse rhizome 40 minutes under tap water repeatedly.After drying surface moisture,
On superclean bench, cut away high about 4～6 centimetres of stem, stay about 1～2 centimetre of high stem with rhizome, dip 70% wine with cotton balls
Essence wipes rhizome and about 1～2 centimetre of high stem surface, is finally sterilized 15 minutes with 0.1% mercuric chloride solution, aquesterilisa rinses 5 times,
Cut away the stem apart from high about 1～2 centimetre of rhizome and stay rhizome, and inoculate rhizome in inducing culture.The induction training of this comparative example
Foster base is all identical with the condition of embodiment 3.Inoculation 1 week about is it has been observed that inducing culture 100% pollutes.
The embodiment of the present invention is as follows with respect to the advantage of comparative example：Due to the difference of explant sterilization method, the present invention is real
Apply example and greatly reduce pollution rate in inducing culture for the explant, thus improve work efficiency.
Claims (8)
1. a kind of quick breeding method for tissue culture of Rhizoma Stahlianthi Involucrati is it is characterised in that comprise the following steps：
The selection of (a) explant：The rhizome taking the tender sprouting of eugonic Rhizoma Stahlianthi Involucrati children is explant；
The sterilization of (b) explant：Explant tap water is cleaned, blade cut by shears, stay rhizome and high by 5～7 apart from rhizome
Centimetre stem, carry out the sterilization of explant；Cut away the stem apart from high 1～2 centimetre of rhizome and stay rhizome, and inoculate rhizome to induction training
Cultivate in foster base；
The induction of (c) Multiple Buds：Culture on inducing culture is until explant forms adventitious bud；Described inducing culture becomes
It is divided into MS, 6 benzyl aminoadenine 1.0～3.0mg/L, naphthalene acetic acid 0.01～0.10mg/L, coconut juice 10.0～15.0%, sucrose
20～30g/L, carrageenan 9.5～10g/L；
(d) proliferation and subculture：The adventitious bud inducing is cut away and cuts again after partial blade, successive transfer culture enters in proliferated culture medium
The propagation of row adventitious bud；Described proliferation and subculture medium component is MS, 6 benzyl aminoadenine 3.0～8.0mg/L, naphthalene acetic acid
0.1～0.20mg/L, coconut juice 10.0～15.0%, sucrose 20～30g/L, carrageenan 9.5～10g/L；
(e) root culture：When 3～5 centimetres of subculture height of seedling, cut and be inoculated in root media, described root media
Composition is MS, naphthalene acetic acid 0.1～1.0mg/L, coconut juice 10.0～15.0%, sucrose 20～30g/L, carrageenan 9.5～10g/L；
(f) acclimatization and transplantses：When 5～8 centimetres of the 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, the Bravo solution soaking with 0.1%～0.3% 30～40 minutes, be transplanted to by Huang
In the substrate of mud, perlite and peat soil mixing, keep ventilated, humidity 70 85%, temperature at 20 32 DEG C, natural light
Obtain transplanted seedling according to after CMC model.
2. Rhizoma Stahlianthi Involucrati according to claim 1 quick breeding method for tissue culture it is characterised in that：Step (a) is described
Explant be Rhizoma Stahlianthi Involucrati (Stahlianthus involucratus) the tender sprouting of 5～20 centimetres of high childrens rhizome.
3. Rhizoma Stahlianthi Involucrati according to claim 1 quick breeding method for tissue culture it is characterised in that：With mass percent
Densitometer, the sterilization of described explant is first to soak rhizome 30～40 minutes with 0.1% 0.2% potassium permanganate solutions, then uses
0.1%～0.3% Bravo and thiophanate-methyl mixed solution, soak rhizome 40～50 minutes, then anti-under tap water
Rinse rhizome 30～40 minutes again；After drying surface moisture, on superclean bench, cut away high 4～6 centimetres of stem, stay 1～2
Centimetre high stem with rhizome, dips 70% alcohol wipe rhizome and 1～2 centimetre high of stem surface with cotton balls, finally with 0.1%
Mercuric chloride solution is sterilized 15～20 minutes, and aquesterilisa rinses 4～5 times.
4. Rhizoma Stahlianthi Involucrati according to claim 2 quick breeding method for tissue culture it is characterised in that：Described Bravo and
The volume ratio of thiophanate-methyl is 1：1.
5. Rhizoma Stahlianthi Involucrati according to claim 2 quick breeding method for tissue culture it is characterised in that：Described yellow mud, treasure
The volume ratio of Zhu Yan and peat soil is 1：1：1.
6. Rhizoma Stahlianthi Involucrati according to claim 2 quick breeding method for tissue culture it is characterised in that：In step (5) such as
Fruit temperature is higher than 32 DEG C, with blower fan and cascade cooling.
7. Rhizoma Stahlianthi Involucrati according to claim 2 quick breeding method for tissue culture it is characterised in that：Described inducing culture
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.
8. Rhizoma Stahlianthi Involucrati according to claim 2 quick breeding method for tissue culture it is characterised in that：Step (b), step
C the temperature of (), step (d) and the described culture of step (e) is 25～30 DEG C, intensity of illumination is 2000～2300lx, light application time
It is controlled to 12 hours/day.
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Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
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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 |
| CN107593443A (en) * | 2017-10-18 | 2018-01-19 | 广东省农业科学院环境园艺研究所 | A kind of quick breeding method for tissue culture of little Hua kaempferia galamgas |
| 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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Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| 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 |
| CN107593443A (en) * | 2017-10-18 | 2018-01-19 | 广东省农业科学院环境园艺研究所 | A kind of quick breeding method for tissue culture of little Hua kaempferia galamgas |
| 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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