CN103493733B - Method for quickly inducing generation of fritillaria cirrhosa bulb - Google Patents
Method for quickly inducing generation of fritillaria cirrhosa bulb Download PDFInfo
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- CN103493733B CN103493733B CN201310437399.5A CN201310437399A CN103493733B CN 103493733 B CN103493733 B CN 103493733B CN 201310437399 A CN201310437399 A CN 201310437399A CN 103493733 B CN103493733 B CN 103493733B
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Abstract
The invention discloses a method for quickly inducing generation of fritillaria cirrhosa bulb, belonging to the field of biotechnology. In the method provided by the invention, a tissue culture bulb is selected to be put in MS+KT1-3mg.L-1+NAA0.1-1mg.L-1 culture medium, after the tissue culture bulb is cultured for 30-40 days at the culture temperature of 12-18 DEG C without illumination, succulent juvenile leaves of fritillaria cirrhosa generated by the tissue culture bulb are obtained, then, the succulent juvenile leaves of fritillaria cirrhosa are put in a bulb generation inducing culture medium for cultivation, thus, regeneration of fritillaria cirrhosa bulb on the succulent juvenile leaves of fritillaria cirrhosa is realized, and a new way for quick and massive generation of medicinal parts of fritillaria cirrhosa is provided. The method has simple steps and strong operability, has the characteristics of low investment, low cost and high efficiency, and has important practical significance for solving the problem of shortage of fritillaria cirrhosa medicinal materials.
Description
Technical field
The invention belongs to biological technical field, relate to a kind of method that rapid induction Bulbus Fritillariae Cirrhosae bulb produces particularly.
Background technology
Bulbus Fritillariae Cirrhosae (Fritillaria cirrhosa D.Don) is Liliaceae herbaceos perennial, is used as medicine, is famous and precious Genuine crude drugs in Sichuan, can be rated as the treasured in medicine with bulb.Bulbus Fritillariae Cirrhosae plant is to growing environment requirement harshness, and it likes cold cloudy weather conditions, has cold-resistant, to like wet, like concealment, fearness high temperature characteristic.Temperature reaches 30 DEG C or ground temperature more than 25 DEG C, and plant will wither, and can not survive in the area that, temperature low at height above sea level is high, so Bulbus Fritillariae Cirrhosae could normal growth at the height of height above sea level 3500m ~ 4500m.Bulbus Fritillariae Cirrhosae is the good medicine of important treatment cough, and for lung-heat type cough, the few phlegm of dry cough, deficiency of Yin labor is coughed, and coughs up the symptoms such as sputum streaked with blood.Because Bulbus Fritillariae Cirrhosae is expensive, excavating in addition in recent years without plan blindness, natural resources is increasingly exhausted, estimates also to be difficult to alleviate within nearly decades.Tissue culture technique growth cycle is short, reproduction rate is high, break away from the Nature the adverse effect of the four seasons, change round the clock and Catastrophe climate, and condition is homogeneous, very favourable to plant growth, be convenient to stably to carry out cultivate in the anniversary and produce, it is one of effective ways solving Bulbus Fritillariae Cirrhosae herb resource shortage problem that the method therefore applying tissue cultures carries out Bulbus Fritillariae Cirrhosae Fast-propagation.Existing employing Bulbus Fritillariae Cirrhosae Plant Leaf is that the generation of explant induction bulb all will produce acquisition bulb through the callus tissue culture stage again.As everyone knows, tissue cultures material in the chromosomal variation more easily produced through callus phase cell, thus is unfavorable for the genetic stability guaranteeing cultivation bulb medicinal material.
Summary of the invention
The object of the invention is to overcome the deficiencies in the prior art, provide a kind of method that rapid induction Bulbus Fritillariae Cirrhosae bulb produces, the method step is simple, and workable, process stabilizing, is applicable to the extensive use in industrialized production.
The object of the invention is to be achieved through the following technical solutions:
The method that rapid induction Bulbus Fritillariae Cirrhosae bulb produces, it comprises the following steps:
S1: the group training bulb of diameter at 0.5 ~ 2cm choosing induction generation, access MS+KT 1 ~ 3mgL
-1+ NAA 0.1 ~ 1 mgL
-1cultivate in medium, condition of culture is: cultivation temperature is cultivate 30 ~ 40 days under 12 ~ 18 DEG C and non-illuminated conditions, namely obtains the Bulbus Fritillariae Cirrhosae carnification spire produced by group training bulb;
S2: choose the carnification spire that spire upper end is white or yellow-white, after it being cut from leaf sheath base, accesses the medium MS+6-BA 1 ~ 3mgL of induction bulb generation in the mode kept flat
-1+ TDZ 0.1 ~ 0.5mgL
-1+ IAA 0.1 ~ 1mgL
-1+ Ad 10 ~ 25mgL
-1in cultivate, condition of culture is: culture materials is placed on be 4 ~ 8 DEG C and under the condition of culture of unglazed photograph cultivate 5 ~ 10 days;
S3: the culture materials after S2 step process being placed on cultivation temperature is 15 ~ 28 DEG C, illumination every day 4 ~ 8h, intensity of illumination is cultivate 20 ~ 40 days under the condition of culture of 200 ~ 600lx.
Preferably, the group training bulb described in S1 step is directly 1.2cm, access MS+KT 2mgL
-1+ NAA 0.5mgL
-1medium in.
Preferably, the cultivation temperature described in S1 step is 15 DEG C.
Preferably, the condition of culture described in S1 step is cultivate 35 days under non-illuminated conditions.
Preferably, the medium that the induction bulb described in S2 step produces is MS+6-BA 2mgL
-1+ TDZ 0.3mgL
-1+ IAA 0.5mgL
-1+ Ad 18mgL
-1.
Preferably, the cultivation temperature described in S2 step is: 6 DEG C, and cultivates 7 days under the condition of culture of unglazed photograph.
Preferably, the cultivation temperature described in S3 step is: 20 DEG C, and every day, light application time was 6h, and intensity of illumination is cultivate 30 days under the condition of culture of 400lx.
Beneficial effect of the present invention is:
Present invention achieves Direct Regeneration Bulbus Fritillariae Cirrhosae bulb from Bulbus Fritillariae Cirrhosae meat spire, the rapid batch for Bulbus Fritillariae Cirrhosae medicinal part produces and provides a new way.The method can significantly improve the sprout rate of bulb, obviously shortens the output that crop cycle improves Bulbus Fritillariae Cirrhosae.The method step is simple, and workable, process stabilizing, is applicable to the extensive use in industrialized production.
Embodiment
Below in conjunction with embodiment, the present invention will be further described, and protection scope of the present invention is not limited to the following stated:
Embodiment 1: a kind of method that rapid induction Bulbus Fritillariae Cirrhosae bulb produces, it comprises the following steps:
S1: the group training bulb of diameter at 0.5 ~ 2cm choosing induction generation, access MS+KT 1 ~ 3mgL
-1+ NAA 0.1 ~ 1mgL
-1cultivate in medium, condition of culture is: cultivation temperature is cultivate 30 ~ 40 days under 12 ~ 18 DEG C and non-illuminated conditions, namely obtains the Bulbus Fritillariae Cirrhosae carnification spire produced by group training bulb;
S2: choose the carnification spire that spire upper end is white or yellow-white, after it being cut from leaf sheath base, accesses the medium MS+6-BA 1 ~ 3mgL of induction bulb generation in the mode kept flat
-1+ TDZ 0.1 ~ 0.5mgL
-1+ IAA 0.1 ~ 1mgL
-1+ Ad 10 ~ 25mgL
-1in cultivate, condition of culture is: culture materials is placed on be 4 ~ 8 DEG C and under the condition of culture of unglazed photograph cultivate 5 ~ 10 days;
S3: the culture materials after S2 step process being placed on cultivation temperature is 15 ~ 28 DEG C, illumination every day 4 ~ 8h, intensity of illumination is cultivate 20 ~ 40 days under the condition of culture of 200 ~ 600lx.
Embodiment 2: a kind of method that rapid induction Bulbus Fritillariae Cirrhosae bulb produces, it comprises the following steps:
S1: the group training bulb of diameter at 1.2cm choosing induction generation, access MS+KT 2mgL
-1+ NAA 0.5mgL
-1cultivate in medium, condition of culture is: cultivation temperature is cultivate 35 days under 15 DEG C and non-illuminated conditions, namely obtains the Bulbus Fritillariae Cirrhosae carnification spire produced by group training bulb;
S2: choose the carnification spire that spire upper end is white or yellow-white, after it being cut from leaf sheath base, accesses the medium MS+6-BA 2mgL of induction bulb generation in the mode kept flat
-1+ TDZ 0.3mgL
-1+ IAA 0.5mgL
-1+ Ad 18mgL
-1in cultivate, condition of culture is: culture materials is placed on be 6 DEG C and under the condition of culture of unglazed photograph cultivate 7 days;
S3: the culture materials after S2 step process being placed on cultivation temperature is 20 DEG C, illumination every day 6h, intensity of illumination is cultivate 30 days under the condition of culture of 400lx.
Embodiment 3: a kind of method that rapid induction Bulbus Fritillariae Cirrhosae bulb produces, it comprises the following steps:
S1: the group training bulb of diameter at 0.5cm choosing induction generation, access MS+KT 1mgL
-1+ NAA 0.1mgL
-1cultivate in medium, condition of culture is: cultivation temperature is cultivate 30 days under 12 DEG C and non-illuminated conditions, namely obtains the Bulbus Fritillariae Cirrhosae carnification spire produced by group training bulb;
S2: choose the carnification spire that spire upper end is white or yellow-white, after it being cut from leaf sheath base, accesses the medium MS+6-BA 1mgL of induction bulb generation in the mode kept flat
-1+ TDZ 0.1mgL
-1+ IAA 0.1mgL
-1+ Ad 10mgL
-1in cultivate, condition of culture is: culture materials is placed on be 4 DEG C and under the condition of culture of unglazed photograph cultivate 5 days;
S3: the culture materials after S2 step process being placed on cultivation temperature is 15 DEG C, illumination every day 4h, intensity of illumination is cultivate 20 days under the condition of culture of 200lx.
Embodiment 4: a kind of method that rapid induction Bulbus Fritillariae Cirrhosae bulb produces, it comprises the following steps:
S1: the group training bulb of diameter at 2cm choosing induction generation, access MS+KT 3mgL
-1+ NAA 1mgL
-1cultivate in medium, condition of culture is: cultivation temperature is cultivate 40 days under 18 DEG C and non-illuminated conditions, namely obtains the Bulbus Fritillariae Cirrhosae carnification spire produced by group training bulb;
S2: choose the carnification spire that spire upper end is white or yellow-white, after it being cut from leaf sheath base, accesses the medium MS+6-BA 3mgL of induction bulb generation in the mode kept flat
-1+ TDZ 0.5mgL
-1+ IAA 1mgL
-1+ Ad 25mgL
-1in cultivate, condition of culture is: culture materials is placed on be 8 DEG C and under the condition of culture of unglazed photograph cultivate 10 days;
S3: the culture materials after S2 step process being placed on cultivation temperature is 28 DEG C, illumination every day 8h, intensity of illumination is cultivate 40 days under the condition of culture of 600lx.
Embodiment 5: a kind of method that rapid induction Bulbus Fritillariae Cirrhosae bulb produces, it comprises the following steps:
S1: the group training bulb of diameter at 1cm choosing induction generation, access MS+KT 2mgL
-1+ NAA 1mgL
-1cultivate in medium, condition of culture is: cultivation temperature is cultivate 35 days under 15 DEG C and non-illuminated conditions, namely obtains the Bulbus Fritillariae Cirrhosae carnification spire produced by group training bulb;
S2: choose the carnification spire that spire upper end is white or yellow-white, after it being cut from leaf sheath base, accesses the medium MS+6-BA 2mgL of induction bulb generation in the mode kept flat
-1+ TDZ 0.3mgL
-1+ IAA 0.5mgL
-1+ Ad 18mgL
-1in cultivate, condition of culture is: culture materials is placed on be 6 DEG C and under the condition of culture of unglazed photograph cultivate 7 days;
S3: the culture materials after S2 step process being placed on cultivation temperature is 18 DEG C, illumination every day 6h, intensity of illumination is cultivate 35 days under the condition of culture of 400lx.
Embodiment 6: a kind of method that rapid induction Bulbus Fritillariae Cirrhosae bulb produces, it comprises the following steps:
S1: the group training bulb of diameter at 0.6cm choosing induction generation, access MS+KT 1.5mgL
-1+ NAA 0.6mgL
-1cultivate in medium, condition of culture is: cultivation temperature is cultivate 30 days under 15 DEG C and non-illuminated conditions, namely obtains the Bulbus Fritillariae Cirrhosae carnification spire produced by group training bulb;
S2: choose the carnification spire that spire upper end is white or yellow-white, after it being cut from leaf sheath base, accesses the medium MS+6-BA 1.5mgL of induction bulb generation in the mode kept flat
-1+ TDZ 0.5mgL
-1+ IAA 0.6mgL
-1+ Ad 18mgL
-1in cultivate, condition of culture is: culture materials is placed on be 7 DEG C and under the condition of culture of unglazed photograph cultivate 8 days;
S3: the culture materials after S2 step process being placed on cultivation temperature is 18 DEG C, illumination every day 8h, intensity of illumination is cultivate 20 days under the condition of culture of 300lx.
Embodiment 7: a kind of method that rapid induction Bulbus Fritillariae Cirrhosae bulb produces, it comprises the following steps:
S1: the group training bulb of diameter at 1.5cm choosing induction generation, access MS+KT 2mgL
-1+ NAA 0.5 mgL
-1cultivate in medium, condition of culture is: cultivation temperature is cultivate 35 days under 16 DEG C and non-illuminated conditions, namely obtains the Bulbus Fritillariae Cirrhosae carnification spire produced by group training bulb;
S2: choose the carnification spire that spire upper end is white or yellow-white, after it being cut from leaf sheath base, accesses the medium MS+6-BA 2mgL of induction bulb generation in the mode kept flat
-1+ TDZ 0.3mgL
-1+ IAA 0.5mgL
-1+ Ad 18mgL
-1in cultivate, condition of culture is: culture materials is placed on be 6 DEG C and under the condition of culture of unglazed photograph cultivate 7 days;
S3: the culture materials after S2 step process being placed on cultivation temperature is 20 DEG C, illumination every day 6h, intensity of illumination is cultivate 20 ~ 40 days under the condition of culture of 400lx.
Embodiment 8: a kind of method that rapid induction Bulbus Fritillariae Cirrhosae bulb produces, it comprises the following steps:
S1: the group training bulb of diameter at 2cm choosing induction generation, access MS+KT 2mgL
-1+ NAA0.5 mgL
-1cultivate in medium, condition of culture is: cultivation temperature is cultivate 35 days under 16 DEG C and non-illuminated conditions, namely obtains the Bulbus Fritillariae Cirrhosae carnification spire produced by group training bulb;
S2: choose the carnification spire that spire upper end is white or yellow-white, after it being cut from leaf sheath base, accesses the medium MS+6-BA 2mgL of induction bulb generation in the mode kept flat
-1+ TDZ 0.3mgL
-1+ IAA 0.5mgL
-1+ Ad 20mgL
-1in cultivate, condition of culture is: culture materials is placed on be 6 DEG C and under the condition of culture of unglazed photograph cultivate 8 days;
S3: the culture materials after S2 step process being placed on cultivation temperature is 25 DEG C, illumination every day 7h, intensity of illumination is cultivate 40 days under the condition of culture of 500lx.
Embodiment 9: a kind of method that rapid induction Bulbus Fritillariae Cirrhosae bulb produces, it comprises the following steps:
S1: the group training bulb of diameter at 0.5 ~ 2cm choosing induction generation, access MS+KT 1 ~ 3mgL
-1+ NAA 0.1 ~ 1 mgL
-1cultivate in medium, condition of culture is: cultivation temperature is cultivate 35 days under 15 DEG C and non-illuminated conditions, namely obtains the Bulbus Fritillariae Cirrhosae carnification spire produced by group training bulb;
S2: choose the carnification spire that spire upper end is white or yellow-white, after it being cut from leaf sheath base, accesses the medium MS+6-BA 1 ~ 3mgL of induction bulb generation in the mode kept flat
-1+ TDZ 0.1 ~ 0.5mgL
-1+ IAA 0.1 ~ 1mgL
-1+ Ad 10 ~ 25mgL
-1in cultivate, condition of culture is: culture materials is placed on be 6 DEG C and under the condition of culture of unglazed photograph cultivate 7 days;
S3: the culture materials after S2 step process being placed on cultivation temperature is 20 DEG C, illumination every day 6h, intensity of illumination is cultivate 20 ~ 40 days under the condition of culture of 400lx.
Effect of the present invention is further illustrated below by experiment:
Experiment one: the impact that the inventive method occurs bulb
S1 step: the group training bulb of diameter at 1.2 cm choosing induction generation, access MS+KT 2mgL
-1+ NAA 0.5mgL
-1in medium, be 15 DEG C in cultivation temperature and after 35 days, obtain the Bulbus Fritillariae Cirrhosae carnification spire produced by group training bulb with cultivation under non-illuminated conditions.S2 step: choose the carnification spire that spire upper end is white, after it being cut from leaf sheath base, accesses the medium MS+6-BA 2mgL of induction bulb generation in the mode kept flat
-1+ TDZ 0.3mgL
-1+ IAA 0.5mgL
-1+ Ad 18mgL
-1in cultivate, condition of culture is: culture materials is placed on be 6 DEG C and under the condition of culture of unglazed photograph cultivate 7 days.S3 step: the culture materials after S2 step process being placed on cultivation temperature is 22 DEG C, illumination every day 6h, intensity of illumination is cultivate 30 days under the condition of culture of 400lx.The generation bulb of Bulbus Fritillariae Cirrhosae carnification spire 100%, the bulb number that average each spire explant produces is 15 ~ 30.
Experiment two: change the medium that induction group training bulb sprouts fast into MS+6-BA 0.5mgL in experiment one S2 step
-1+ Cefotaxime Sodium 100mgL
-1in medium, other step is with experiment one; After cultivating through 45 days, the sprout rate of group training bulb is 58.62%.
Experiment three: in experiment one S2 step, the condition of culture that sprouts fast of induction group training bulb is changed into cultivation temperature be 28 DEG C, after illumination every day 24 hours, intensity of illumination cultivate 45 days under being the condition of culture of 2500lx, the sprout rate of group training bulb is 89.18%.
Experiment four: change the medium that inducing embryoid body produces into MS+ Cefotaxime Sodium 500mgL in experiment one S3 step
-1, other step is with experiment one; After cultivating through 50 days, the inductivity of its embryoid is 58.81%, and the embryoid shape produced mostly is elongated taper shape.
Experiment five: the medium that sprouted fast by group training bulb in experiment one S2 step changes MS0 minimal medium into, and namely do not add any phytohormone and antibiotic, other step is with experiment one; After cultivating through 45 days, the sprout rate of group training bulb is only 13.17%.
Experiment six: by experiment one S2 step, the condition of culture that sprouts fast of group training bulb is changed into cultivation temperature be 15 DEG C, cultivate 45 days under the condition of unglazed photograph after, other step is with experiment one; The sprout rate of group training bulb is 11.85%.
Experiment seven: in experiment one S3 step, take the yellow green blade-section not cutting spire top, other step is with experiment one, and after cultivating through 50 days, the inductivity of its embryoid is 38.71%.
Experiment eight: in experiment one S3 step, directly inserted in medium just to connect mode (the morphology lower end with leaf sheath explant) by the white leaf sheath of spire base portion, other step is with experiment one, and after cultivating through 50 days, the inductivity of its embryoid is 28.56%.
Experiment nine: in experiment one S3 step, directly inserted in medium by the white leaf sheath of spire base portion in reversal connection mode (the morphology upper end with leaf sheath explant), other step is with real experiment one, and after cultivating through 50 days, the inductivity of its embryoid is 39.16%.
Show by experiment, the inventive method achieves Direct Regeneration Bulbus Fritillariae Cirrhosae bulb from Bulbus Fritillariae Cirrhosae meat spire, and the rapid batch for Bulbus Fritillariae Cirrhosae medicinal part produces and provides a new way.The method can significantly improve the sprout rate of bulb, obviously shortens crop cycle, and then improves the output of Bulbus Fritillariae Cirrhosae.
Claims (7)
1. a method for rapid induction Bulbus Fritillariae Cirrhosae bulb generation, is characterized in that: it comprises the following steps:
S1: the group training bulb of diameter at 0.5 ~ 2cm choosing induction generation, access MS+KT 1 ~ 3mgL
-1+ NAA 0.1 ~ 1 mgL
-1cultivate in medium, condition of culture is: cultivation temperature is cultivate 30 ~ 40 days under 12 ~ 18 DEG C and non-illuminated conditions, namely obtains the Bulbus Fritillariae Cirrhosae carnification spire produced by group training bulb;
S2: choose the carnification spire that spire upper end is white or yellow-white, after it being cut from leaf sheath base, accesses the medium MS+6-BA 1 ~ 3mgL of induction bulb generation in the mode kept flat
-1+ TDZ 0.1 ~ 0.5mgL
-1+ IAA 0.1 ~ 1mgL
-1+ Ad 10 ~ 25mgL
-1in cultivate, condition of culture is: culture materials is placed on be 4 ~ 8 DEG C and under the condition of culture of unglazed photograph cultivate 5 ~ 10 days;
S3: the culture materials after S2 step process being placed on cultivation temperature is 15 ~ 28 DEG C, illumination every day 4 ~ 8h, intensity of illumination is cultivate 20 ~ 40 days under the condition of culture of 200 ~ 600lx.
2. the method for a kind of rapid induction Bulbus Fritillariae Cirrhosae bulb generation according to claim 1, is characterized in that: the group training bulb diameter described in S1 step is 1.2cm, access MS+KT 2mgL
-1+ NAA 0.5mgL
-1medium in.
3. the method for a kind of rapid induction Bulbus Fritillariae Cirrhosae bulb generation according to claim 1, is characterized in that: the cultivation temperature described in S1 step is 15 DEG C.
4. the method for a kind of rapid induction Bulbus Fritillariae Cirrhosae bulb generation according to claim 1, is characterized in that: the condition of culture described in S1 step is cultivate 35 days under non-illuminated conditions.
5. the method for a kind of rapid induction Bulbus Fritillariae Cirrhosae bulb generation according to claim 1, is characterized in that: the medium that the induction bulb described in S2 step produces is MS+6-BA 2mgL
-1+ TDZ 0.3mgL
-1+ IAA 0.5mgL
-1+ Ad 18mgL
-1.
6. the method for a kind of rapid induction Bulbus Fritillariae Cirrhosae bulb generation according to claim 1, is characterized in that: the cultivation temperature described in S2 step is: 6 DEG C, and cultivates 7 days under the condition of culture of unglazed photograph.
7. the method for a kind of rapid induction Bulbus Fritillariae Cirrhosae bulb generation according to claim 1, it is characterized in that: the cultivation temperature described in S3 step is: 20 DEG C, every day, light application time was 6h, and intensity of illumination is cultivate 30 days under the condition of culture of 400lx.
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| CN114847159A (en) * | 2022-04-28 | 2022-08-05 | 云南龙藏生物科技有限公司 | Tissue culture regeneration method for bulbus fritillariae cirrhosae |
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| CN104957037B (en) * | 2015-07-07 | 2017-03-22 | 甘肃省科学院生物研究所 | Method for cultivating fritillaria przewalskii test-tube bulblet |
| CN105519435B (en) * | 2016-01-06 | 2017-10-13 | 南京海源中药饮片有限公司 | A kind of tissue culture method of induction fritillaria thunbergii clove |
| CN111296211A (en) * | 2020-03-12 | 2020-06-19 | 重庆市药物种植研究所 | Rapid propagation method of fritillaria taipaiensis bulbs |
| CN111492976B (en) * | 2020-05-18 | 2022-09-13 | 成都大学 | Method for culturing fleshy straight roots of bulbus fritillariae cirrhosae |
| CN111492977B (en) * | 2020-05-18 | 2022-09-13 | 成都大学 | Induced culture method of bulbus fritillariae cirrhosae round bulbs |
| CN112243631B (en) * | 2020-09-14 | 2022-03-22 | 云南省农业科学院花卉研究所 | Method for rapidly breaking dormancy of green flower lily seed bulbs |
| CN112931210B (en) * | 2021-03-18 | 2022-06-21 | 四川迪菲特药业有限公司 | Bulb proliferation culture method taking fritillaria paniculata bulb discs as explants |
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| KR100586495B1 (en) * | 2004-11-30 | 2006-06-07 | 삼성에버랜드 주식회사 | A method of embryogenic callus formation with a high frequency from bulb scale of tulip and shoot regeneration from the embryogenic callus |
| CN102002474B (en) * | 2009-09-03 | 2012-08-08 | 薛永新 | Rapid bulb multiplication method by using fritillaria cirrhosa leaves as explant |
| CN103238456B (en) * | 2013-05-31 | 2015-09-16 | 太仓市天合新材料科技有限公司 | A kind of method utilizing Bulbus Fritillariae Cirrhosae scale Fast-propagation Bulbus Fritillariae Cirrhosae |
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| CN114847159A (en) * | 2022-04-28 | 2022-08-05 | 云南龙藏生物科技有限公司 | Tissue culture regeneration method for bulbus fritillariae cirrhosae |
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