CN102017894A - In vitro rapid propagation method of Stephania epigaea - Google Patents
In vitro rapid propagation method of Stephania epigaea Download PDFInfo
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- CN102017894A CN102017894A CN 201010508270 CN201010508270A CN102017894A CN 102017894 A CN102017894 A CN 102017894A CN 201010508270 CN201010508270 CN 201010508270 CN 201010508270 A CN201010508270 A CN 201010508270A CN 102017894 A CN102017894 A CN 102017894A
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Abstract
The invention provides an in vitro rapid propagation method of Stephania epigaea. The method comprises the following steps of: firstly carrying out explant selection, explant sterilization, adventitious bud induction culture, adventitious bud propagation culture, subculture, rooting culture, transition transplantation and the like on various corresponding culture media, and directly transplanting after acclimatization. A variety introducing rate reaches 90%, and a transplantation survival rate is over 85%. By using the method, the problems of poor propagation capacity and slow propagation speed of the Stephania epigaea by using a conventional method are solved, and the sterile propagation technique system of the Stephania epigaea is established. The purposes of high variety introducing efficiency and rapid proliferation speed are attained. The mass and standard production of the fine variety seedlings of the Stephania epigaea is achieved.
Description
Technical field
The present invention relates to a kind of rapid breeding method of plant, especially the method for in-vitro rapid propagation of mountain tortoise belongs to field of plant growing technology.
Background technology
The mountain tortoise is China's traditional Chinese medicine, mainly is distributed in ground such as Yunnan Province of China, Guizhou, Sichuan.Because its meat piece root is quaint, peculiar, fruit is exquisite, and the leaf look delicate and pretty, and plant profile uniqueness is lovely, therefore, except as the medicinal material, also can be used as the pot plant that has ornamental value, and perhaps plant is used in gardens, greening.The tool medical value of mountain tortoise plant be its very large root piece, be rich in alkaloid in its composition, appearance along with new isolation technics and highly sensitive detecting instrument, a large amount of alkaloids are separated, content is up to 3%～4%, be widely used in anti-inflammation, calmness, regulating qi-flowing for relieving pain etc. clinically, mountain tortoise root piece is the main production raw material of Rotundine simultaneously.Along with medical value and the ornamental value of mountain tortoise are constantly excavated, market is also strengthened during to the demand of mountain tortoise gradually.Yet because the mountain tortoise is a dioecism, and seed is little, and the kind shell is hard, sprouts comparatively difficulty under field conditions (factors), so the fertility extreme difference, adds that being subjected to the people in recent years is the such environmental effects that causes, and sharply reduces population of mountain tortoise and individual amount.Only, far can not satisfy the growing market demand by traditional sowing, block division method.Therefore, the vegetative propagation system that cost is low, the time is short, survival rate is high that needs research and development to make new advances to enlarge mountain tortoise breeding amount, realizes the industrialization production of mountain tortoise.
Summary of the invention
The object of the present invention is to provide tissue culture and the method for quickly breeding of a kind of mountain tortoise, setting up mountain tortoise population, enlarge mountain tortoise breeding amount, improve the rate of increase and rooting rate, for scale, the industrialization production of mountain tortoise provides high quality seedling.
For achieving the above object, the invention provides the method for in-vitro rapid propagation of a kind of like this mountain tortoise, it is characterized in that through following processing step:
A, the selection mountain tortoise piece root that volume is very large, shape is graceful, eye is more, plant in indoor perlite to growing young sprout, and downcut this young sprout as explant, remove blade, be cut into the long stem section of 2～3cm, clean the stem section with the conventional water that is added with an amount of washing agent, clean with clear water again, with mass concentration is 0.1% mercuric chloride solution sterilization, 1～4min, with rinsed with sterile water 3～4 times, aseptic stem section;
B, the aseptic stem section of A step is inoculated in the following inducing culture:
The MS medium
6-benzamido group purine (6-BA) 0.5～1.0mg/L
2,4 dichlorophenoxyacetic acid (2,4-D) 0～0.05mg/L
Methyl (NAA) 0～0.05mg/L
Agar 6.5g/L
White sugar 30g/L
PH value 5.8～6.0
In cultivation temperature is 22～28 ℃, and intensity of illumination is 1500～2000Lx, and light application time is under the condition of 8～12h/d, and inducing culture 16～30 days sprouts 1～4 indefinite bud in stem segment base portion;
C, the indefinite bud of B step is downcut, is connected in the following proliferated culture medium:
The MS medium
6-benzamido group purine (6-BA) 1.0～2.0mg/L
2,4 dichlorophenoxyacetic acid (2,4-D) 0.02～0.04mg/L
Methyl (NAA) 0.02～0.04mg/L
Agar 6.5g/L
White sugar 30g/L
PH value 5.8～6.0
In cultivation temperature is 22～28 ℃, intensity of illumination is 2000～2500Lx, light application time is under the condition of 8～12h/d, enrichment culture 20～25 days, 1～9 propagation of the new propagation of each indefinite bud bud, downcutting newly-increased height is the propagation bud of 2～5cm, is seeded in the new proliferated culture medium identical with this step, the condition of culture identical by this step continues enrichment culture, and so successive transfer culture is to the propagation bud that obtains q.s;
D, the propagation bud of C step is cut into the long stem section of 2～3cm by joint, is seeded in the following successive transfer culture:
3-indolebutyric acid (IBA) 0.1～0.3mg/L
Methyl (NAA) 0.1～0.3mg/L
Agar 6.5g/L
White sugar 30g/L
PH value 5.8～6.0
Under the condition of culture identical, carry out strong seedling culture, get the seedling of growing thickly healthy and strong, the band leaf with the C step;
E, the seedling of growing thickly of D step is divided into individual plant, inserts in the following root media:
The 1/2MS medium
6-benzamido group purine (6-BA) 0～0.2mg/L
3-indolebutyric acid (IBA) 0～1.0mg/L
Methyl (NAA) 0～1.0mg/L
Agar 7.0g/L
White sugar 30g/L
PH value 5.8～6.0
In cultivation temperature is 22～28 ℃, and intensity of illumination is 2000～2500Lx, and light application time is under the condition of 8～12h/d, cultivates the seedling of must taking root 15～20 days;
F, the seedling of taking root of E step was taken exercise under outdoor scattered light 5～7 days, get seedling, clean medium on the seedling with clear water, put into 0.1% carbendazim solution and sterilized 1～2 minute, transplant to laterite is housed: river sand: in the perlite=Seedling bag of 1: 1: 1 quality than matrix, and sprinkle an amount of humus soil in stromal surface, water routinely, the management of fertilising, the extermination of disease and insect pest, survival rate reaches more than 85%, when seedling grows to 3～5cm height, and when bearing a large amount of fibrous root, promptly obtain kind planting seedlings.
The present invention has following advantage and effect:
1, adopt such scheme, can carry out fast breeding the stem section of mountain tortoise, keep simultaneously and fixed that mountain tortoise piece root is very large, good strains of seeds such as high yield, shape are peculiar, thereby when guaranteeing medicinal demand, can satisfy the specific demand of ornamental value again;
2, source of seedling is single, is easy to control, has solved the very difficult difficult problem that mountain tortoise merit is fixed or kept getting off of conventional seed growing system, has improved seedling quality effectively;
3, can effectively shorten the production cycle, reproduction speed is fast, and 20～25d is an one-period, and breeding rate reaches 1～9, is easy to scale, standardization and the batch production production of high quality seedling;
4, utilize tissue culture technique realization anniversary in culturing room to produce, saved land resources, improved economic benefit, overcome the difficult point that seedling can't be produced in the anniversary;
5, the present invention is optimized integration to the key link in the tortoise seedling production technology flow process of mountain, take low hormone combination, hormone is used with and take turns the mode of usefulness, reach the purpose that improves the rate of increase and reduce variation, the propagation seedling of being produced and the seedling of taking root, seedling strain stalwartness, internode is short and sturdy, the leaf look dark green, Functionality, quality and appealing design, transplanting survival rate is up to more than 85%;
6, the sterile rootage seedling direct transplantation enters in the Seedling bag, has saved the link that the tissue cultivating seedling nutritive cube is transplanted, and the seedling of transplant survival is convenient to transportation, can directly plant after plucking Seedling bag.
Embodiment
For flesh and blood of the present invention is described better, below in conjunction with embodiments of the invention, but content of the present invention is not limited in these.
Embodiment 1
A, the selection mountain tortoise piece root that volume is very large, shape is graceful, eye is more, plant in indoor perlite to growing young sprout, and downcut this young sprout as explant, remove blade, be cut into the long stem section of 2～3cm, clean the stem section with the conventional water that is added with an amount of washing agent, clean with clear water again, with mass concentration is 0.1% mercuric chloride solution sterilization 1min, with rinsed with sterile water 3 times, aseptic stem section;
B, the aseptic stem section of A step is inoculated in the following inducing culture:
The MS medium
6-benzamido group purine (6-BA) 0.5mg/L
Agar 6.5g/L
White sugar 30g/L
PH value 5.8
In cultivation temperature is 22 ℃, and intensity of illumination is 1500Lx, and light application time is under the condition of 12h/d, and inducing culture 16 days is sprouted in stem segment base portion and be there emerged a indefinite bud, minimum sends a bud, maximum sends four buds;
C, the indefinite bud of B step is downcut, is connected in the following proliferated culture medium:
The MS medium
6-benzamido group purine (6-BA) 1.0mg/L
2,4 dichlorophenoxyacetic acid (2,4-D) 0.02mg/L
Methyl (NAA) 0.02mg/L
Agar 6.5g/L
White sugar 30g/L
PH value 5.8
In cultivation temperature is 22 ℃, intensity of illumination is 2000Lx, light application time is under the condition of 12h/d, enrichment culture 20 days, and each indefinite bud increases out the propagation bud again newly, bud of minimum propagation, nine buds of maximum propagation, downcutting newly-increased height is the propagation bud of 2～5cm, is seeded in the new proliferated culture medium identical with this step, the condition of culture identical by this step continues enrichment culture, and so successive transfer culture is to the propagation bud that obtains q.s;
D, the propagation bud of C step is cut into the long stem section of 2～3cm by joint, is seeded in the following subculture medium:
3-indolebutyric acid (IBA) 0.1mg/L
Methyl (NAA) 0.1mg/L
Agar 6.5g/L
White sugar 30g/L
PH value 5.8
Under the condition of culture identical, carry out strong seedling culture, get the seedling of growing thickly healthy and strong, the band leaf with the C step;
E, the seedling of growing thickly of D step is divided into individual plant, inserts in the following root media, the bud seedling is too high, the seedling point that can downcut long 2～3cm is forwarded in the following root media, remaining base portion returns in the proliferated culture medium that is inoculated into the C step, proceeds enrichment culture, and described root media is:
The 1/2MS medium
Agar 7.0g/L
White sugar 30g/L
PH value 5.8
In cultivation temperature is 22 ℃, and intensity of illumination is 2000Lx, and light application time is under the condition of 12h/d, cultivates the seedling of must taking root 15 days;
F, the seedling of taking root of E step was taken exercise 5 days under outdoor scattered light, get seedling, clean medium on the seedling with clear water, put into 0.1% carbendazim solution sterilization 1 minute, transplant to laterite is housed: river sand: in the perlite=Seedling bag of 1: 1: 1 quality than matrix, and sprinkle an amount of humus soil in stromal surface, water routinely, the management of fertilising, the extermination of disease and insect pest, survival rate reaches more than 85%, when seedling grows to 3～5cm height, and when bearing a large amount of fibrous root, the kind that promptly directly is planted in big Tanaka is planted seedlings.
Embodiment 2
A, select that volume is very large, graceful special, the more mountain tortoise piece root of eye of shape, plant in indoor perlite to growing young sprout, and downcut this young sprout as explant, remove blade, be cut into the long stem section of 2～3cm, clean the stem section with the conventional water that is added with an amount of washing agent, clean with clear water again, with mass concentration is 0.1% mercuric chloride solution sterilization 4min, with rinsed with sterile water 4 times, aseptic stem section;
B, the aseptic stem section of A step is inoculated in the following inducing culture:
The MS medium
6-benzamido group purine (6-BA) 1.0mg/L
2,4 dichlorophenoxyacetic acid (2,4-D) 0.05mg/L
Methyl (NAA) 0.05mg/L
Agar 6.5g/L
White sugar 30g/L
PH value 6.0
In cultivation temperature is 28 ℃, and intensity of illumination is 2000Lx, and light application time is under the condition of 8h/d, inducing culture 30 days, and indefinite bud is sprouted by portion at the stem segment base, a minimum bud, four maximum buds;
C, the indefinite bud of B step is downcut, is connected in the following proliferated culture medium:
The MS medium
6-benzamido group purine (6-BA) 2.0mg/L
2,4 dichlorophenoxyacetic acid (2,4-D) 0.04mg/L
Methyl (NAA) 0.04mg/L
Agar 6.5g/L
White sugar 30g/L
PH value 6.0
In cultivation temperature is 28 ℃, intensity of illumination is 2500Lx, light application time is under the condition of 8h/d, enrichment culture 25 days, and each indefinite bud increases out the propagation bud newly, a minimum bud, nine maximum buds, downcutting newly-increased height is the propagation bud of 2～5cm, is seeded in the new proliferated culture medium identical with this step, the condition of culture identical by this step continues enrichment culture, and so successive transfer culture is to the propagation bud that obtains q.s;
D, the propagation bud of C step is cut into the long stem section of 2～3cm by joint, is seeded in the following successive transfer culture:
3-indolebutyric acid (IBA) 0.3mg/L
Methyl (NAA) 0.3mg/L
Agar 6.5g/L
White sugar 30g/L
PH value 6.0
Under the condition of culture identical, carry out strong seedling culture, get the seedling of growing thickly healthy and strong, the band leaf with the C step;
E, the seedling of growing thickly of D step is divided into individual plant, inserts in the following root media, the bud seedling is too high, the seedling point that can downcut long 2～3cm is forwarded in the following root media, remaining base portion returns in the proliferated culture medium that is inoculated into the C step, proceeds enrichment culture, and described root media is:
The 1/2MS medium
6-benzamido group purine (6-BA) 0.2mg/L
3-indolebutyric acid (IBA) 1.0mg/L
Methyl (NAA) 1.0mg/L
Agar 7.0g/L
White sugar 30g/L
PH value 6.0
In cultivation temperature is 28 ℃, and intensity of illumination is 2500Lx, and light application time is under the condition of 8h/d, cultivates the seedling of must taking root 20 days;
F, the seedling of taking root of E step was taken exercise 7 days under outdoor scattered light, get seedling, clean medium on the seedling with clear water, put into 0.1% carbendazim solution sterilization 2 minutes, transplant to laterite is housed: river sand: in the perlite=Seedling bag of 1: 1: 1 quality than matrix, and sprinkle an amount of humus soil in stromal surface, water routinely, the management of fertilising, the extermination of disease and insect pest, survival rate reaches more than 85%, when seedling grows to 3～5cm height, and when bearing a large amount of fibrous root, the kind that promptly directly is planted in big Tanaka is planted seedlings.
Embodiment 3
A, select that volume is very large, graceful peculiar, the mountain tortoise piece root that eye is more of shape, plant in indoor perlite to growing young sprout, and downcut this young sprout as explant, remove blade, be cut into the long stem section of 2～3cm, clean the stem section with the conventional water that is added with an amount of washing agent, clean with clear water again, with mass concentration is 0.1% mercuric chloride solution sterilization 2min, with rinsed with sterile water 3 times, aseptic stem section;
B, the aseptic stem section of A step is inoculated in the following inducing culture:
The MS medium
6-benzamido group purine (6-BA) 0.8mg/L
Methyl (NAA) 0.03mg/L
Agar 6.5g/L
White sugar 30g/L
PH value 5.9
In cultivation temperature is 25 ℃, and intensity of illumination is 1800Lx, and light application time is under the condition of 10h/d, inducing culture 22 days, and indefinite bud is sprouted by portion at the stem segment base, and few is a bud, and many is four buds;
C, the indefinite bud of B step is downcut, is connected in the following proliferated culture medium:
The MS medium
6-benzamido group purine (6-BA) 1.5mg/L
2,4 dichlorophenoxyacetic acid (2,4-D) 0.03mg/L
Methyl (NAA) 0.03mg/L
Agar 6.5g/L
White sugar 30g/L
PH value 5.9
In cultivation temperature is 25 ℃, and intensity of illumination is 2200Lx, and light application time is under the condition of 10h/d, enrichment culture 22 days, and each indefinite bud increases out the propagation bud newly.Minimum is a bud, maximum is nine buds, and downcutting newly-increased height is the propagation bud of 2～5cm, is seeded in the new proliferated culture medium identical with this step, the condition of culture identical by this step continues enrichment culture, and so successive transfer culture is to the propagation bud that obtains q.s;
D, the propagation bud of C step is cut into the long stem section of 2～3cm by joint, is seeded in the following successive transfer culture:
3-indolebutyric acid (IBA) 0.2mg/L
Methyl (NAA) 0.2mg/L
Agar 6.5g/L
White sugar 30g/L
PH value 5.9
Under the condition of culture identical, carry out strong seedling culture, get the seedling of growing thickly healthy and strong, the band leaf with the C step;
E, the seedling of growing thickly of D step is divided into individual plant, inserts in the following root media, the bud seedling is too high, the seedling point that can downcut long 2～3cm is forwarded in the following root media, remaining base portion returns in the proliferated culture medium that is inoculated into the C step, proceeds enrichment culture, and described root media is:
The 1/2MS medium
6-benzamido group purine (6-BA) 0.1mg/L
3-indolebutyric acid (IBA) 0.5mg/L
Agar 7.0g/L
White sugar 30g/L
PH value 5.9
In cultivation temperature is 25 ℃, and intensity of illumination is 2300Lx, and light application time is under the condition of 10h/d, cultivates the seedling of must taking root 18 days;
F, the seedling of taking root of E step was taken exercise 6 days under outdoor scattered light, get seedling, clean medium on the seedling with clear water, put into 0.1% carbendazim solution sterilization 1 minute, transplant to laterite is housed: river sand: in the perlite=Seedling bag of 1: 1: 1 quality than matrix, and sprinkle an amount of humus soil in stromal surface, water routinely, the management of fertilising, the extermination of disease and insect pest, survival rate reaches more than 85%, when seedling grows to 3～5cm height, and when bearing a large amount of fibrous root, the kind that promptly directly is planted in big Tanaka is planted seedlings.
Embodiment 4
A, the selection mountain tortoise piece root that volume is very large, shape is graceful, eye is more, plant in indoor perlite to growing young sprout, and downcut this young sprout as explant, remove blade, be cut into the long stem section of 2～3cm, clean the stem section with the conventional water that is added with an amount of washing agent, clean with clear water again, with mass concentration is 0.1% mercuric chloride solution sterilization 3min, with rinsed with sterile water 4 times, aseptic stem section;
B, the aseptic stem section of A step is inoculated in the following inducing culture:
The MS medium
6-benzamido group purine (6-BA) 0.6mg/L
2,4 dichlorophenoxyacetic acid (2,4-D) 0.02mg/L
Agar 6.5g/L
White sugar 30g/L
PH value 5.8
In cultivation temperature is 26 ℃, and intensity of illumination is 1800Lx, and light application time is under the condition of 9h/d, inducing culture 20 days, and indefinite bud is sprouted by portion at the stem segment base, a minimum bud, four maximum buds;
C, the indefinite bud of B step is downcut, is connected in the following proliferated culture medium:
The MS medium
6-benzamido group purine (6-BA) 2.0mg/L
2,4 dichlorophenoxyacetic acid (2,4-D) 0.02mg/L
Methyl (NAA) 0.04mg/L
Agar 6.5g/L
White sugar 30g/L
PH value 5.8
In cultivation temperature is 26 ℃, intensity of illumination is 2300Lx, light application time is under the condition of 9h/d, enrichment culture 23 days, and each indefinite bud increases out the propagation bud newly, a minimum bud, nine maximum buds, downcutting newly-increased height is the propagation bud of 2～5cm, is seeded in the new proliferated culture medium identical with this step, the condition of culture identical by this step continues enrichment culture, and so successive transfer culture is to the propagation bud that obtains q.s;
D, the propagation bud of C step is cut into the long stem section of 2～3cm by joint, is seeded in the following successive transfer culture:
3-indolebutyric acid (IBA) 0.1mg/L
Methyl (NAA) 0.3mg/L
Agar 6.5g/L
White sugar 30g/L
PH value 5.8
Under the condition of culture identical, carry out strong seedling culture, get the seedling of growing thickly healthy and strong, the band leaf with the C step;
E, the seedling of growing thickly of D step is divided into individual plant, inserts in the following root media, the bud seedling is too high, the seedling point that can downcut long 2～3cm is forwarded in the following root media, remaining base portion returns in the proliferated culture medium that is inoculated into the C step, proceeds enrichment culture, and described root media is:
The 1/2MS medium
Methyl (NAA) 1.0mg/L
Agar 7.0g/L
White sugar 30g/L
PH value 5.8
In cultivation temperature is 26 ℃, and intensity of illumination is 2300Lx, and light application time is under the condition of 9h/d, cultivates the seedling of must taking root 18 days;
F, the seedling of taking root of E step was taken exercise 6 days under outdoor scattered light, get seedling, clean medium on the seedling with clear water, put into 0.1% carbendazim solution sterilization 2 minutes, transplant to laterite is housed: river sand: in the perlite=Seedling bag of 1: 1: 1 quality than matrix, and sprinkle an amount of humus soil in stromal surface, water routinely, the management of fertilising, the extermination of disease and insect pest, survival rate reaches more than 85%, when seedling grows to 3～5cm height, and when bearing a large amount of fibrous root, the kind that promptly directly is planted in big Tanaka is planted seedlings.
Claims (1)
1. the method for in-vitro rapid propagation of a mountain tortoise is characterized in that through following processing step:
A, the selection mountain tortoise piece root that volume is very large, shape is graceful, eye is more, plant in indoor perlite to growing young sprout, and downcut this young sprout as explant, remove blade, be cut into the long stem section of 2～3cm, clean the stem section with the conventional water that is added with an amount of washing agent, clean with clear water again, with mass concentration is 0.1% mercuric chloride solution sterilization, 1～4min, with rinsed with sterile water 3～4 times, aseptic stem section;
B, the aseptic stem section of A step is inoculated in the following inducing culture:
The MS medium
6-benzamido group purine (6-BA) 0.5～1.0mg/L
2,4 dichlorophenoxyacetic acid (2,4-D) 0～0.05mg/L
Methyl (NAA) 0～0.05mg/L
Agar 6.5g/L
White sugar 30g/L
PH value 5.8～6.0
In cultivation temperature is 22～28 ℃, and intensity of illumination is 1500～2000Lx, and light application time is under the condition of 8～12h/d, and inducing culture 16～30 days sprouts 1～4 indefinite bud in stem segment base portion;
C, the indefinite bud of B step is downcut, is connected in the following proliferated culture medium:
The MS medium
6-benzamido group purine (6-BA) 1.0～2.0mg/L
2,4 dichlorophenoxyacetic acid (2,4-D) 0.02～0.04mg/L
Methyl (NAA) 0.02～0.04mg/L
Agar 6.5g/L
White sugar 30g/L
PH value 5.8～6.0
In cultivation temperature is 22～28 ℃, intensity of illumination is 2000～2500Lx, light application time is under the condition of 8～12h/d, enrichment culture 20～25 days, 1～9 propagation of the new propagation of each indefinite bud bud, downcutting newly-increased height is the propagation bud of 2～5cm, is seeded in the new proliferated culture medium identical with this step, the condition of culture identical by this step continues enrichment culture, and so successive transfer culture is to the propagation bud that obtains q.s;
D, the propagation bud of C step is cut into the long stem section of 2～3cm by joint, is seeded in the following successive transfer culture:
3-indolebutyric acid (IBA) 0.1～0.3mg/L
Methyl (NAA) 0.1～0.3mg/L
Agar 6.5g/L
White sugar 30g/L
PH value 5.8～6.0
Under the condition of culture identical, carry out strong seedling culture, get the seedling of growing thickly healthy and strong, the band leaf with the C step;
E, the seedling of growing thickly of D step is divided into individual plant, inserts in the following root media:
The 1/2MS medium
6-benzamido group purine (6-BA) 0～0.2mg/L
3-indolebutyric acid (IBA) 0～1.0mg/L
Methyl (NAA) 0～1.0mg/L
Agar 7.0g/L
White sugar 30g/L
PH value 5.8～6.0
In cultivation temperature is 22～28 ℃, and intensity of illumination is 2000～2500Lx, and light application time is under the condition of 8～12h/d, cultivates the seedling of must taking root 15～20 days;
F, the seedling of taking root of E step was taken exercise under outdoor scattered light 5～7 days, get seedling, clean medium on the seedling with clear water, put into 0.1% carbendazim solution and sterilized 1～2 minute, transplant to laterite is housed: river sand: in the perlite=Seedling bag of 1: 1: 1 quality than matrix, and sprinkle an amount of humus soil in stromal surface, water routinely, the management of fertilising, the extermination of disease and insect pest, survival rate reaches more than 85%, when seedling grows to 3～5cm height, and when bearing a large amount of fibrous root, promptly obtain kind planting seedlings.
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| CN105210882A (en) * | 2015-11-05 | 2016-01-06 | 广西壮族自治区药用植物园 | A kind of tissue culture propagation of Diels Stephania Root |
| CN109122318A (en) * | 2018-08-29 | 2019-01-04 | 西南林业大学 | A kind of Stephania epigaea tissue culture propagation method |
| CN109496852A (en) * | 2018-11-27 | 2019-03-22 | 钟天路 | A kind of tissue culture technique of mountain tortoise |
| CN109964813A (en) * | 2017-12-28 | 2019-07-05 | 云南生物谷药业股份有限公司 | A kind of tissue culture mating system of high cycleanine and the Stephania epigaea of cepharanthine content |
| CN110250004A (en) * | 2019-07-18 | 2019-09-20 | 四川迪菲特药业有限公司 | A kind of method of mountain tortoise tissue-culturing quick-propagation |
| CN115349446A (en) * | 2022-10-19 | 2022-11-18 | 云南植虫药生物科技有限公司 | Method for producing cepharanthine by using tortoise callus |
| CN117717003A (en) * | 2024-02-02 | 2024-03-19 | 云南省农业科学院药用植物研究所 | Tissue culture rapid propagation method and application of anoectochilus formosanus combined with bottle external rooting technology |
-
2010
- 2010-10-15 CN CN 201010508270 patent/CN102017894A/en active Pending
Cited By (10)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN105210882A (en) * | 2015-11-05 | 2016-01-06 | 广西壮族自治区药用植物园 | A kind of tissue culture propagation of Diels Stephania Root |
| CN109964813A (en) * | 2017-12-28 | 2019-07-05 | 云南生物谷药业股份有限公司 | A kind of tissue culture mating system of high cycleanine and the Stephania epigaea of cepharanthine content |
| CN109122318A (en) * | 2018-08-29 | 2019-01-04 | 西南林业大学 | A kind of Stephania epigaea tissue culture propagation method |
| CN109496852A (en) * | 2018-11-27 | 2019-03-22 | 钟天路 | A kind of tissue culture technique of mountain tortoise |
| CN110250004A (en) * | 2019-07-18 | 2019-09-20 | 四川迪菲特药业有限公司 | A kind of method of mountain tortoise tissue-culturing quick-propagation |
| CN110250004B (en) * | 2019-07-18 | 2021-12-14 | 四川迪菲特药业有限公司 | Tissue culture and rapid propagation method for Stephania delavayi Diels |
| CN115349446A (en) * | 2022-10-19 | 2022-11-18 | 云南植虫药生物科技有限公司 | Method for producing cepharanthine by using tortoise callus |
| CN115349446B (en) * | 2022-10-19 | 2022-12-27 | 云南省农业科学院药用植物研究所 | Method for producing cepharanthine by using tortoise callus |
| CN117717003A (en) * | 2024-02-02 | 2024-03-19 | 云南省农业科学院药用植物研究所 | Tissue culture rapid propagation method and application of anoectochilus formosanus combined with bottle external rooting technology |
| CN117717003B (en) * | 2024-02-02 | 2024-04-30 | 云南省农业科学院药用植物研究所 | Tissue culture rapid propagation method and application of anoectochilus formosanus combined with bottle external rooting technology |
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