CN103444526B - A kind of growth-promoting cultivation method of Fritillaria unibracteata - Google Patents
A kind of growth-promoting cultivation method of Fritillaria unibracteata Download PDFInfo
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Abstract
The invention discloses a kind of growth-promoting cultivation method of Fritillaria unibracteata, comprise the step such as process, Initial culture, squamous subculture, culture of rootage, rooting tube plantlet greenhouse acclimatization and transplants, soil matrix management of the selection of material and sterilization, material, the present invention adopts fresh Fritillaria unibracteata bulb ball to breed as explant, the availability of bulb ball is very high, reach more than 95%, and the plant branches and leaves cultivated are luxuriant, well developed root system, and plant strain growth is vigorous; A diameter 4cm bulb ball approximately can breed 150-200 clove, and limited resource namely can be utilized to breed, and reproduction coefficient is very high.In Subculture after Calli Differentiation goes out thin bud, raise cultivation temperature, improve periodicity of illumination and intensity of illumination, increase soil fertility, Pest management rate, shorten incubation time like this.
Description
Technical field
The present invention relates to the method for tissue culture of a kind of bulb of fritillary, particularly relate to a kind of method for quickly breeding being applicable to Fritillaria unibracteata.
Background technology
Fritillaria unibracteata (F.unibiacteataHsiaoetK.C.Hsia) is traditional Chinese medicine famousr and precious in Fritillaria, its bulb can effectively cough-relieving, eliminate the phlegm, clearing heat and moistening lung, be particularly suitable for the elderly and the cough of children's cureless intractable cold of insufficiency type.The demand of Fritillaria unibracteata in Chinese Medicinal Materials Markets is high especially, but its bulb growth speed is slow especially, the time that commodity medicinal material generally needs 4 years is grown up to from seed, due to bulb of fritillary good drug efficacy, be worth high, add that artificial planting difficulty is large, reproduction coefficient is low, growth cycle is long, therefore commodity price of medicinal material remains high always, and demand is increasing in recent years, causes the price of Fritillaria unibracteata also more and more higher.Wild Fritillaria unibracteata is mainly distributed in the of short duration area growth of the loud and sonorous cold of physical features, sunshine strong, air oxygen detrition, spring and autumn.Main producing region Ruoergai, Hongyuan two county are because affecting by landform, cold current etc., long-term without the summer, this climatic characteristic constitutes the special environment condition of Fritillaria unibracteata growth, so this special growing environment determine Fritillaria unibracteata field and artificial planting more difficult, in order to expand the breeding of Fritillaria unibracteata, tissue culture technique is utilized to carry out Fast-propagation very necessary.At present the research of the tissue culture technique of the bulb of fritillary is begun to take shape, but be not also a lot of to the research of Fritillaria unibracteata, and propagation method is not also on a large scale, does not form the breeding system of complete set.
Summary of the invention
The technical problem that the present invention mainly solves is to provide a kind of method utilizing tissue culture technique to breed Fritillaria unibracteata, and the fast breeding for Fritillaria unibracteata provides the tissue culture propagating system of complete set.
For solving the problems of the technologies described above, the technical scheme that the present invention adopts is:
A growth-promoting cultivation method for Fritillaria unibracteata, is characterized in that, comprise the following steps:
(1) selection of material and sterilization: first select robust growth, ripe fresh Fritillaria unibracteata bulb ball, peel off scale and the remaining old root of base portion of bulb outermost layer shriveling, put into clear water after running water 30min and soak 15min, after 50% carbendazim, 500 times of immersion bubble 20-30min, by the strip off in layer of bulb ball, 20min is irradiated again under uviol lamp, then superclean bench is proceeded to, superclean bench is first used the alcohol immersion 10 seconds of 70-75%, then aseptic water washing is used, proceed to again dripped 2 ~ 3 POLYSORBATE 80s 0.1% mercuric chloride solution in soak 5min, with aseptic water washing 4 times,
(2) process of material: the scale aseptic filter paper of disinfecting is blotted, aseptically also has base section to isolate according to upper, middle and lower portion material, be cut into the fritter of 4-6mm respectively;
(3) Initial culture: under sterile conditions the explant that step (2) obtains is seeded on Initial culture base, this medium is the ZT of NAA, 0.4mg/L at 6-BA, the 1.2mg/L that with the addition of 1.0mg/L in MS conventional medium; Cultivation temperature is 26 DEG C, light application time 14h/d, and intensity of illumination 2200Lx, PH are 6.5, agar 5g/L, forms callus through 15-20 days;
(4) squamous subculture: the callus of Initial culture is cut and is seeded on subculture medium, this medium is the sucrose of KT and 30mg/L of NAA, the 0.3mg/L of 6-BA, the 0.4mg/L that with the addition of 0.5mg/L in 3/4MS conventional medium, cultivation temperature is 22 DEG C, periodicity of illumination 13h/d, luminous intensity is 2500lux, PH6.5, after 4-5 days time differentiated thin bud, raise cultivation temperature to 25 DEG C, periodicity of illumination is 14h/d, luminous intensity is 2800lux, continues to cultivate the clove that just can differentiate cluster cluster for 6-10 days;
(5) culture of rootage: differentiation clove is out inoculated in following inducing culture one by one respectively and carries out root induction: the KT of IBA, 0.4mg/L of the active carbon of MS medium, 1.5mg/L, the agar of 6g/L, 0.3mg/L, cultivation temperature is 23 DEG C, periodicity of illumination 12h/d, luminous intensity is 2000lux, pH value is 6.0, treated that clove grew to 3cm through 20-25 days high, when root system is more healthy and stronger and when growing 6-10 root, namely start the next stage;
(5) soil matrix management: regulate soil pH value to be 7.0 ~ 8.2, use after mixing with 5 times of volume fine earths after Cosan is sieved, except regulating soil pH value with Cosan, Cosan is then better with peat composed of rotten mosses mixing application effect, or use pine needle to coordinate Cosan to improve soil, wherein pine needle, original soil, muck, Cosan are according to ratio of weight and number 2:3:1:2, or apply in soil saw not, the peat composed of rotten mosses, liver moss and Cosan carry out soil melioration, wherein saw not, the peat composed of rotten mosses, liver moss and Cosan ratio of weight and number be 1:1:1:2;
Increase soil organic matter content: the field planting ditch digging wide 50cm, dark 40cm, organic matter, fertilizer, original soil are improved the soil in the ratio of 1:1:1, organic matter uses the acid peat composed of rotten mosses, if first January more than must be stacked with sawdust, bark, become thoroughly decomposed completely after decomposing and re-use, as employment domestic animals manure become thoroughly decomposed or straw leaf rot to become thoroughly decomposed after the natural fertilizers that formed, note not mixing ash, lime alkaline matter, content reaches more than 3% ~ 5%, to increase the permeability of soil;
Soil covers: can carry out after soil covers seedling planting, by covering uniform fold at bed surface, wide 1 meter, thick 5 ~ 10 centimetres, cover 2 cm thicks more every year later, to keep original thickness, the new sawdust decomposed if application is not rotted, the nitrogenous fertilizer of 50% need be enriched, rot the sawdust decomposed, nitrogen fertilizer amount should correspondingly reduce, cover unregistered land film and can prevent soil water evaporation, control weeds, improve ground temperature, preferably the orchard of drip irrigation facility is being had to apply, cover unregistered land film and cover the organic matter better or front weed killer herbicide of field planting of effect that combines and kill weeds, entirely cultivate field after field planting and sow green manure Trifolium repense, the leguminous plants such as red clover, field can be covered completely after March, soil fertility can be increased again,
(6) rooting tube plantlet greenhouse acclimatization and transplants:
By the rearmounted greenhouse temperature of test-tube plantlet uncork of taking root at 25-28 DEG C, humidity 80-85%, intensity of illumination after 3-5 days, washes the medium on seedling at 5000-5500lx condition lower refining seedling off, is transplanted in the soil matrix in above-mentioned steps (5); When extraneous nocturnal temperature is at 10-15 DEG C, when day temperature is between 18-25, test-tube plantlet is transplanted, in 5 days of the initial stage of transplanting, with covered rearing with plastic film, to keep humidity more than 80%, and controlled light intensity is at 2700-3300lx, throws off film afterwards gradually and suitably increase illumination, until under transplanted seedling is placed in natural conditions.
The invention has the beneficial effects as follows:
(1) the present invention adopts fresh Fritillaria unibracteata bulb ball to breed as explant, and the availability of bulb ball is very high, reaches more than 95%, and the plant branches and leaves cultivated are luxuriant, well developed root system, and plant strain growth is vigorous; A diameter 4cm bulb ball approximately can breed 150-200 clove, and limited resource namely can be utilized to breed, and reproduction coefficient is very high.
(2) in Subculture after Calli Differentiation goes out thin bud, raise cultivation temperature, improve periodicity of illumination and intensity of illumination, shorten incubation time like this.
(3) in rooting tube plantlet greenhouse acclimatization and transplants process when extraneous nocturnal temperature is at 10-15 DEG C, transplant when day temperature is between 18-25, be conducive to growing of Fritillaria unibracteata, namely the time of transplanting has been held, improve soil fertility, control damage by disease and insect, make the transplanting success rate of test-tube plantlet reach more than 95%.
Embodiment
Below preferred embodiment of the present invention is described in detail, can be easier to make advantages and features of the invention be readily appreciated by one skilled in the art, thus more explicit defining is made to protection scope of the present invention.
embodiment 1
A growth-promoting cultivation method for Fritillaria unibracteata, comprising:
(1) selection of material and sterilization: first select robust growth, ripe fresh Fritillaria unibracteata bulb ball, peel off scale and the remaining old root of base portion of bulb outermost layer shriveling, put into clear water after running water 30min and soak 15min, after 50% carbendazim, 500 times of immersion bubble 20min, by the strip off in layer of bulb ball, 20min is irradiated again under uviol lamp, then superclean bench is proceeded to, superclean bench is first used the alcohol immersion 10 seconds of 70%, then aseptic water washing is used, proceed to again dripped 2 POLYSORBATE 80s 0.1% mercuric chloride solution in soak 5min, with aseptic water washing 4 times,
(2) process of material: the scale aseptic filter paper of disinfecting is blotted, aseptically also has base section to isolate according to upper, middle and lower portion material, be cut into the fritter of 4mm respectively;
(3) Initial culture: under sterile conditions the explant that step (2) obtains is seeded on Initial culture base, this medium is the ZT of NAA, 0.4mg/L at 6-BA, the 1.2mg/L that with the addition of 1.0mg/L in MS conventional medium; Cultivation temperature is 26 DEG C, light application time 14h/d, and intensity of illumination 2200Lx, PH are 6.5, agar 5g/L, forms callus through 15-20 days;
(4) squamous subculture: the callus of Initial culture is cut and is seeded on subculture medium, this medium is the sucrose of KT and 30mg/L of NAA, the 0.3mg/L of 6-BA, the 0.4mg/L that with the addition of 0.5mg/L in 3/4MS conventional medium, cultivation temperature is 22 DEG C, periodicity of illumination 13h/d, luminous intensity is 2500lux, PH6.5, after 4 day time differentiated thin bud, raise cultivation temperature to 25 DEG C, periodicity of illumination is 14h/d, luminous intensity is 2800lux, continues the clove that cultivation just can differentiate cluster cluster for 8 days;
(5) culture of rootage: differentiation clove is out inoculated in following inducing culture one by one respectively and carries out root induction: the KT of IBA, 0.4mg/L of the active carbon of MS medium, 1.5mg/L, the agar of 6g/L, 0.3mg/L, cultivation temperature is 23 DEG C, periodicity of illumination 12h/d, luminous intensity is 2000lux, pH value is 6.0, treated that clove grew to 3cm through 20-25 days high, when root system is more healthy and stronger and when growing 6-10 root, namely start the next stage;
(6) rooting tube plantlet greenhouse acclimatization and transplants: by the rearmounted greenhouse temperature of test-tube plantlet uncork of taking root at 25 DEG C, humidity 80%, intensity of illumination is at 5000lx condition lower refining seedling after 5 days, wash the medium on seedling off, being transplanted to by sheep excrement, humus soil, perlitic mixed proportion is on the seedbed of 1: 3: 2.When extraneous nocturnal temperature is at 10-15 DEG C, when day temperature is between 18-25, test-tube plantlet is transplanted, in 5 days of the initial stage of transplanting, with covered rearing with plastic film, to keep humidity more than 80%, and controlled light intensity is at 2700lx, throws off film afterwards gradually and suitably increase illumination, until under transplanted seedling is placed in natural conditions.
Through the cultivation of above-mentioned steps, the Callus formation rate of Fritillaria unibracteata reaches 95%, and proliferation times reaches 5.8 times, and rooting rate reaches 95%, and final survival rate reaches 88%, and the Quality and yield of Fritillaria unibracteata is all greatly improved.
embodiment 2
A growth-promoting cultivation method for Fritillaria unibracteata, comprises the following steps:
(1) selection of material and sterilization: first select robust growth, ripe fresh Fritillaria unibracteata bulb ball, peel off scale and the remaining old root of base portion of bulb outermost layer shriveling, put into clear water after running water 30min and soak 15min, after 50% carbendazim, 500 times of immersion bubble 30min, by the strip off in layer of bulb ball, 20min is irradiated again under uviol lamp, , then superclean bench is proceeded to, superclean bench is first used the alcohol immersion 10 seconds of 75%, then aseptic water washing is used, proceed to again dripped 3 POLYSORBATE 80s 0.1% mercuric chloride solution in soak 5min, with aseptic water washing 4 times,
(2) process of material: the scale aseptic filter paper of disinfecting is blotted, aseptically also has base section to isolate according to upper, middle and lower portion material, be cut into the fritter of 6mm respectively;
(3) Initial culture: under sterile conditions the explant that step (2) obtains is seeded on Initial culture base, this medium is the ZT of NAA, 0.4mg/L at 6-BA, the 1.2mg/L that with the addition of 1.0mg/L in MS conventional medium; Cultivation temperature is 26 DEG C, light application time 14h/d, and intensity of illumination 2200Lx, PH are 6.5, agar 5g/L, forms callus through 20 days;
(4) squamous subculture: the callus of Initial culture is cut and is seeded on subculture medium, this medium is the sucrose of KT and 30mg/L of NAA, the 0.3mg/L of 6-BA, the 0.4mg/L that with the addition of 0.5mg/L in 3/4MS conventional medium, cultivation temperature is 22 DEG C, periodicity of illumination 13h/d, luminous intensity is 2500lux, PH6.5, after 4-5 days time differentiated thin bud, raise cultivation temperature to 25 DEG C, periodicity of illumination is 14h/d, luminous intensity is 2800lux, continues to cultivate the clove that just can differentiate cluster cluster for 6-10 days;
(5) culture of rootage: differentiation clove is out inoculated in following inducing culture one by one respectively and carries out root induction: the KT of IBA, 0.4mg/L of the active carbon of MS medium, 1.5mg/L, the agar of 6g/L, 0.3mg/L, cultivation temperature is 23 DEG C, periodicity of illumination 12h/d, luminous intensity is 2000lux, pH value is 6.0, treated that clove grew to 3cm through 20-25 days high, when root system is more healthy and stronger and when growing 6-10 root, namely start the next stage;
(6) rooting tube plantlet greenhouse acclimatization and transplants: by the rearmounted greenhouse temperature of test-tube plantlet uncork of taking root at 28 DEG C, humidity 85%, intensity of illumination is at 5500lx condition lower refining seedling after 3 days, wash the medium on seedling off, being transplanted to by sheep excrement, humus soil, perlitic mixed proportion is on the seedbed of 1: 3: 2.When extraneous nocturnal temperature is at 15 DEG C, when day temperature is between 25, test-tube plantlet is transplanted, in 5 days of the initial stage of transplanting, with covered rearing with plastic film, to keep humidity more than 80%, and controlled light intensity is at 3300lx, throws off film afterwards gradually and suitably increase illumination, until under transplanted seedling is placed in natural conditions.
Through the cultivation of above-mentioned steps, the Callus formation rate of Fritillaria unibracteata reaches 92%, and proliferation times reaches 6 times, and rooting rate reaches 98%, and final survival rate reaches 90%.
(5) soil matrix management: regulate soil pH value to be 7.0 ~ 8.2, use after mixing with 5 times of volume fine earths after Cosan is sieved, except regulating soil pH value with Cosan, Cosan is then better with peat composed of rotten mosses mixing application effect, or use pine needle to coordinate Cosan to improve soil, wherein pine needle, original soil, muck, Cosan are according to ratio of weight and number 2:3:1:2, or apply in soil saw not, the peat composed of rotten mosses, liver moss and Cosan carry out soil melioration, wherein saw not, the peat composed of rotten mosses, liver moss and Cosan ratio of weight and number be 1:1:1:2;
Increase soil organic matter content: the field planting ditch digging wide 50cm, dark 40cm, organic matter, fertilizer, original soil are improved the soil in the ratio of 1:1:1, organic matter uses the acid peat composed of rotten mosses, if first January more than must be stacked with sawdust, bark, become thoroughly decomposed completely after decomposing and re-use, as employment domestic animals manure become thoroughly decomposed or straw leaf rot to become thoroughly decomposed after the natural fertilizers that formed, note not mixing ash, lime alkaline matter, content reaches more than 3% ~ 5%, to increase the permeability of soil;
Soil covers: can carry out after soil covers seedling planting, by covering uniform fold at bed surface, wide 1 meter, thick 5 ~ 10 centimetres, cover 2 cm thicks more every year later, to keep original thickness, the new sawdust decomposed if application is not rotted, the nitrogenous fertilizer of 50% need be enriched, rot the sawdust decomposed, nitrogen fertilizer amount should correspondingly reduce, cover unregistered land film and can prevent soil water evaporation, control weeds, improve ground temperature, preferably the orchard of drip irrigation facility is being had to apply, cover unregistered land film and cover the organic matter better or front weed killer herbicide of field planting of effect that combines and kill weeds, entirely cultivate field after field planting and sow green manure Trifolium repense, the leguminous plants such as red clover, field can be covered completely after March, soil fertility can be increased again,
(6) rooting tube plantlet greenhouse acclimatization and transplants:
By the rearmounted greenhouse temperature of test-tube plantlet uncork of taking root at 25-28 DEG C, humidity 80-85%, intensity of illumination after 3-5 days, washes the medium on seedling at 5000-5500lx condition lower refining seedling off, is transplanted in the soil matrix in above-mentioned steps (5); When extraneous nocturnal temperature is at 10-15 DEG C, when day temperature is between 18-25, test-tube plantlet is transplanted, in 5 days of the initial stage of transplanting, with covered rearing with plastic film, to keep humidity more than 80%, and controlled light intensity is at 2700-3300lx, throws off film afterwards gradually and suitably increase illumination, until under transplanted seedling is placed in natural conditions.
Through the cultivation of above-mentioned steps, the Callus formation rate of Fritillaria unibracteata reaches 94%, and proliferation times reaches 7 times, and rooting rate reaches 99%, and final survival rate reaches 98%.
The foregoing is only embodiments of the invention; not thereby the scope of the claims of the present invention is limited; every utilize description of the present invention to do equivalent structure or equivalent flow process conversion; or be directly or indirectly used in other relevant technical fields, be all in like manner included in scope of patent protection of the present invention.
Claims (1)
1. a growth-promoting cultivation method for Fritillaria unibracteata, is characterized in that, comprise the following steps:
(1) selection of material and sterilization: first select robust growth, ripe fresh Fritillaria unibracteata bulb ball, peel off scale and the remaining old root of base portion of bulb outermost layer shriveling, put into clear water after running water 30min and soak 15min, after 50% carbendazim, 500 times of immersion bubble 20-30min, by the strip off in layer of bulb ball, 20min is irradiated again under uviol lamp, then superclean bench is proceeded to, superclean bench is first used the alcohol immersion 10 seconds of 70-75%, then aseptic water washing is used, proceed to again dripped 2 ~ 3 POLYSORBATE 80s 0.1% mercuric chloride solution in soak 5min, with aseptic water washing 4 times,
(2) process of material: the scale aseptic filter paper of disinfecting is blotted, aseptically also has base section to isolate according to upper, middle and lower portion material, be cut into the fritter of 4-6mm respectively;
(3) Initial culture: under sterile conditions the explant that step (2) obtains is seeded on Initial culture base, this medium is the ZT of NAA, the 0.4mg/L of 6-BA, the 1.2mg/L that with the addition of 1.0mg/L in MS conventional medium; Cultivation temperature is 26 DEG C, light application time 14h/d, and intensity of illumination 2200Lx, pH are 6.5, agar 5g/L, forms callus through 15-20 days;
(4) squamous subculture: the callus of Initial culture is cut and is seeded on subculture medium, this medium is the sucrose of KT and 30mg/L of NAA, the 0.3mg/L of 6-BA, the 0.4mg/L that with the addition of 0.5mg/L in 3/4MS conventional medium, cultivation temperature is 22 DEG C, periodicity of illumination 13h/d, luminous intensity is 2500lux, pH6.5, after 4-5 days time differentiated thin bud, raise cultivation temperature to 25 DEG C, periodicity of illumination is 14h/d, luminous intensity is 2800Lx, continues to cultivate the clove differentiating cluster cluster for 6-10 days;
(5) culture of rootage: differentiation clove is out inoculated in following inducing culture one by one respectively and carries out root induction: the KT of the IBA+0.4mg/L of the agar+0.3mg/L of the active carbon+6g/L of MS medium+1.5mg/L, cultivation temperature is 23 DEG C, periodicity of illumination 12h/d, luminous intensity is 2000Lx, pH value is 6.0, treated that clove grew to 3cm through 20-25 days high, when root system is more healthy and stronger and when growing 6-10 root, namely start the next stage;
(6) soil matrix management: regulate soil pH value to be 7.0 ~ 8.2, use after mixing with 5 times of volume fine earths after Cosan is sieved, except regulating soil pH value with Cosan, Cosan is then better with peat composed of rotten mosses mixing application effect, or use pine needle to coordinate Cosan to improve soil, wherein pine needle, original soil, muck, Cosan are according to ratio of weight and number 2:3:1:2, or apply in soil saw not, the peat composed of rotten mosses, liver moss and Cosan carry out soil melioration, wherein saw not, the peat composed of rotten mosses, liver moss and Cosan ratio of weight and number be 1:1:1:2;
Increase soil organic matter content: the field planting ditch digging wide 50cm, dark 40cm, organic matter, fertilizer, original soil are improved the soil in the ratio of 1:1:1, organic matter uses the acid peat composed of rotten mosses, if first January more than must be stacked with sawdust, bark, become thoroughly decomposed completely after decomposing and re-use, as employment domestic animals manure become thoroughly decomposed or straw leaf rot to become thoroughly decomposed after the natural fertilizers that formed, note not mixing ash, lime alkaline matter, content reaches more than 3% ~ 5%, to increase the permeability of soil;
Soil covers: soil carries out after covering seedling planting, by covering uniform fold at bed surface, wide 1 meter, thick 5 ~ 10cm, cover 2cm more every year thick later, to keep original thickness, the new sawdust decomposed if application is not rotted, the nitrogenous fertilizer of 50% need be enriched, rot the sawdust decomposed, nitrogen fertilizer amount should correspondingly reduce, cover unregistered land film and can prevent soil water evaporation, control weeds, improve ground temperature, apply there being the orchard of drip irrigation facility, cover unregistered land film and cover the organic matter better or front weed killer herbicide of field planting of effect that combines and kill weeds, entirely cultivate field after field planting and sow leguminous plant green manure Trifolium repense, red clover, field is covered completely after March, soil fertility can be increased again,
(7) rooting tube plantlet greenhouse acclimatization and transplants:
By the rearmounted greenhouse temperature of test-tube plantlet uncork of taking root at 25-28 DEG C, humidity 80-85%, intensity of illumination after 3-5 days, washes the medium on seedling at 5000-5500Lx condition lower refining seedling off, is transplanted in the soil matrix in above-mentioned steps (6); When extraneous nocturnal temperature is at 10-15 DEG C, when day temperature is between 18-25 DEG C, test-tube plantlet is transplanted, in 5 days of the initial stage of transplanting, with covered rearing with plastic film, to keep humidity more than 80%, and controlled light intensity is at 2700-3300Lx, throws off film afterwards gradually and suitably increase illumination, until under transplanted seedling is placed in natural conditions.
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| Publication number | Priority date | Publication date | Assignee | Title |
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| CN104115745B (en) * | 2014-06-12 | 2016-10-05 | 南京工业大学大丰海洋产业研究院 | A kind of method of explant sterilization in Plant Tissue Breeding |
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Citations (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101161056A (en) * | 2007-09-28 | 2008-04-16 | 中国科学院新疆理化技术研究所 | A method for evoking yili caladium polyploid under culture in vitro |
| CN101213937A (en) * | 2007-12-28 | 2008-07-09 | 浙江省农业科学院 | Method for cultivating detoxification tissue culture bulb of fritillaria thunbergii |
| CN101999317A (en) * | 2009-09-03 | 2011-04-06 | 薛永新 | Multiploid bulb induction method with firtillaria cirrhosa scale leaf as explant |
| CN103238524A (en) * | 2013-05-31 | 2013-08-14 | 常熟市佳盛农业科技发展有限公司 | Method for rapidly breeding fritillaria pallidiflora by using tissue culture technique |
| CN103238525A (en) * | 2013-05-31 | 2013-08-14 | 常熟市佳盛农业科技发展有限公司 | Method for breeding fritillary bulb by tissue culture technique |
| CN103238447A (en) * | 2013-05-24 | 2013-08-14 | 常熟市润丰农业有限公司 | High-quality high-efficiency high-yield blueberry cultivation method |
| CN103238388A (en) * | 2013-05-24 | 2013-08-14 | 常熟市润丰农业有限公司 | Method for scientifically managing soil of newly built blueberry garden |
Family Cites Families (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JPH08112045A (en) * | 1994-10-17 | 1996-05-07 | Koa Oil Co Ltd | Method for mass production of seed and seedling of plant of genus fritillaria |
-
2013
- 2013-08-16 CN CN201310357461.XA patent/CN103444526B/en not_active Expired - Fee Related
Patent Citations (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101161056A (en) * | 2007-09-28 | 2008-04-16 | 中国科学院新疆理化技术研究所 | A method for evoking yili caladium polyploid under culture in vitro |
| CN101213937A (en) * | 2007-12-28 | 2008-07-09 | 浙江省农业科学院 | Method for cultivating detoxification tissue culture bulb of fritillaria thunbergii |
| CN101999317A (en) * | 2009-09-03 | 2011-04-06 | 薛永新 | Multiploid bulb induction method with firtillaria cirrhosa scale leaf as explant |
| CN103238447A (en) * | 2013-05-24 | 2013-08-14 | 常熟市润丰农业有限公司 | High-quality high-efficiency high-yield blueberry cultivation method |
| CN103238388A (en) * | 2013-05-24 | 2013-08-14 | 常熟市润丰农业有限公司 | Method for scientifically managing soil of newly built blueberry garden |
| CN103238524A (en) * | 2013-05-31 | 2013-08-14 | 常熟市佳盛农业科技发展有限公司 | Method for rapidly breeding fritillaria pallidiflora by using tissue culture technique |
| CN103238525A (en) * | 2013-05-31 | 2013-08-14 | 常熟市佳盛农业科技发展有限公司 | Method for breeding fritillary bulb by tissue culture technique |
Non-Patent Citations (2)
| Title |
|---|
| 暗紫贝母愈伤组织和不定芽诱导研究;张波等;《生命科学仪器 》;20110630;第9卷;第48-50页 * |
| 暗紫贝母栽培技术研究;马永贵等;《林业实用技术》;20121231(第4期);第47-78页 * |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105638458A (en) * | 2015-12-29 | 2016-06-08 | 云南澈川生物科技有限公司 | Tissue cultivation method of Fritillaria cirrhosa D.Don |
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