CN105075860A - Herba erigernotis tissue culture seedling cultivation method - Google Patents
Herba erigernotis tissue culture seedling cultivation method Download PDFInfo
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Abstract
The invention relates to a herba erigernotis tissue culture seedling cultivation method and belongs to the technical field of plant tissue culture. The method includes the steps of explant selection, explant sterilization, induction culture, enrichment culture, rooting culture and seedling hardening. Compared with a traditional herba erigernotis seedling production technology, the method has the advantages that the production technology is simple and convenient, the breeding time is short, the reproductive rate is high, and the method is not limited by seasons and places and can be used for industrial production of herba erigernotis seedlings. According to calculation of one herba erigernotis plant, 20-30 explants can be collected from one herba erigernotis plant, calluses can be obtained after 40 days of induction culture, each generation of the calluses is subjected to enrichment culture for 30 days, 6-8 generations are subjected to enrichment culture, rooting culture is conducted for 25 days and the seedlings are hardened for 14-21 days. According to calculation of 6 reproduced generations, 300,000-500,000 commodity seedlings can be obtained within a year through one herba erigernotis adult seedling, and the economic benefits are high.
Description
Technical field
The invention belongs to field of plant tissue culture technique, relate to a kind of important source material medicinal plant method for tissue culture, particularly name is called the medicinal plant method for tissue culture of fleabane flower.
Background technology
Fleabane flower (Erigeron breviscapus), Erigeronbreviscapus (Vant.) Hand.-Mazz., is composite family herbaceos perennial, is also erigeron breviscapus.The high mountain and the Subalpine region that are mainly distributed in height above sea level 1200-3500m open spacious hillside, meadow or border.Pungent, the micro-hardship of herb, temperature, has and dispels rheumatism, activating collaterals to relieve pain, invigorating the spleen disappears long-pending effect, for paralysis, and rheumatic arthritis, toothache, stomachache, infantile malnutrition, polio and meningitis sequela etc. are the important source material plants that China is used for the treatment of angiocardiopathy Chinese patent drug.Fleabane flower medicinal material market demand increases year by year, and limited and excessive the excavating of wild resource, fleabane flower resource reduces just year by year.In recent years the artificial planting technique exploitation of fleabane flower has been carried out in some areas, needs efficiently for artificial planting provides stable, sufficient good seed.Tissue culture technology is adopted to carry out the Fast-propagation of fleabane flower seedling; the scaled artificial planting that can be fleabane flower provides high-quality, stable abundant provenance; can be again the researchs such as the synthesis mechanism of fleabane flower medicinal ingredient, functional gene, molecular breeding to lay the foundation, also can protect the wild resource of wild fleabane flower.
Summary of the invention
The present invention is directed to the deficiency of above-described fleabane flower seedling Traditional breeding processes, it is simple, easy and simple to handle, with short production cycle to have invented a kind of component, directly can produce the tissue culture technique of fleabane flower plant in batches.
The present invention unless otherwise stated, percentage sign representative be mass percent, proportional representation be mass ratio.
The present invention is achieved by the following technical solutions:
A kind of fleabane flower tissue culture method, comprises the steps:
Step (1), the selection of explant: select the fleabane flower tender floral disc of children or axillalry bud as explant;
Step (2), the sterilization of explant: will through the selected explant of step (1), with liquid soap and running water clean clean after, be added to 2-5%(v/v) soak 8-12min in the Tween-80 aqueous solution, then running water shower 2h is used, finally successively with the sterilization of alcohol, clorox and mercuric chloride;
Step (3), Fiber differentiation: after the explant disinfected through step (2) is cut off base portion wound, the inducing culture being seeded in MS+8.0-12.0mg/LIAA+5.0-8.0mg/L6-BA is cultivated, culture environment is temperature 23-25 DEG C, intensity of illumination 1500-2000lux, odd-numbered day light application time is carry out Fiber differentiation under the condition of 12h, obtains Cong Miao;
Step (4), Multiplying culture: the little Cong Cong Miaofen that step (3) obtains being cut to the 2-3 strain of band callus, then the proliferated culture medium being seeded in MS+0.5-1.0mg/LIAA+4.0-7.0mg/LKT is cultivated, culture environment is temperature 23-25 DEG C, intensity of illumination 1500-2000lux, odd-numbered day light application time is carry out Multiplying culture under the condition of 12h, obtains tufted seedling;
Step (5), culture of rootage: divided by tufted seedling and be cut to individual plant, is then seeded on the root media of 1/2MS+0.8-1.2mg/LIAA+2.0g/L active carbon, at 23-25 DEG C, intensity of illumination 1500-2000lux, odd-numbered day light application time is carry out culture of rootage under the environment of 12h, obtains seedling of taking root;
Step (6), hardening: step (5) is obtained take root seedling together with tissue culture bottle be placed in the rate of sheltering from heat or light be 80% green house in carry out in-bottle seeding 2 days, after unscrew bottle cap hardening 1 day again, then seedling removed medium and clean after medium, transplant and plastic covering film, transplanting medium adopts humus soil: perlite=1:0.9-1.1, and after transplanting, air humidity remains on 70%-80%, by 2-3 week cellar culture, namely can be used as commercial seedling.
Further, preferably, the concrete grammar that step (2) middle liquid soap and running water clean is: rinsed by explant liquid soap after washing 10-20min, remains for 3-4 time with tap water to without liquid soap.
Further, preferably, in step (2), the concrete grammar of sterilization is: first use 75%(v/v) alcohol disinfecting 10-12s, again with saturated aqueous sodium hypochlorite solution sterilization 10-20min, after use aseptic water washing 1-2 time, then use aseptic water washing 1-2 time, then use 0.05%(w/v) mercuric chloride solution sterilization 5-10min, finally uses aseptic water washing 4-5 time.
Further, preferably, in step (2), sterilization is carried out on superclean bench.
In technique scheme, the time of the Fiber differentiation described in step (3) is 25-35 days.
In technique scheme, the subculture cycle of the Multiplying culture described in step (4) is 25-35 days.
In technique scheme, the incubation time of the culture of rootage described in step (5) is 25-30 days.
Further, preferably, described fleabane flower tissue culture method, comprises the steps:
Step (1), the selection of explant: fleabane flower is colonizated in booth, management method waters the biological organic fertilizer liquid executed through abundant fermentation maturity routinely, chooses stalwartness, without damage by disease and insect plant, its young tender floral disc of clip or axillalry bud are as explant;
Step (2), the sterilization of explant: the explant will selected through step (1), rinse with liquid soap and wash 10-20min, remain to without liquid soap for 3-4 time with tap water, then explant is added to 2-5%(v/v) soak 8-12min in the Tween-80 aqueous solution, after using running water shower 2h again, be transferred on superclean bench and sterilize; During sterilization, first use 75%(v/v) alcohol disinfecting 10-12s, rear aseptic water washing 1-2 time, again with saturated aqueous sodium hypochlorite solution sterilization 10-20min, then use aseptic water washing 1-2 time, then use 0.05%(w/v) mercuric chloride solution sterilization 5-10min, finally uses aseptic water washing 4-5 time;
Step (3), Fiber differentiation: after the explant disinfected through step (2) is cut off base portion wound, the inducing culture being seeded in MS+8.0-12.0mg/LIAA+5.0-8.0mg/L6-BA is cultivated, culture environment is temperature 23-25 DEG C, intensity of illumination 1500-2000lux, odd-numbered day light application time is carry out Fiber differentiation under the condition of 12h, and after 25-35 days, an one-step inducing goes out the Cong Miao with callus;
Step (4), Multiplying culture: the little Cong Cong Miaofen that step (3) obtains being cut to the 2-3 strain of band callus, then the proliferated culture medium being seeded in MS+0.5-1.0mg/LIAA+4.0-7.0mg/LKT is cultivated, culture environment is temperature 23-25 DEG C, intensity of illumination 1500-2000lux, odd-numbered day light application time is carry out Multiplying culture under the condition of 12h, within 25-35 days, is a subculture cycle, the rate of increase can reach 1:5-1:7, obtains tufted seedling;
Step (5), culture of rootage: tufted seedling is divided and is cut to individual plant, then be seeded on the root media of 1/2MS+0.8-1.2mg/LIAA+2.0g/L active carbon, at 23-25 DEG C, intensity of illumination 1500-2000lux, odd-numbered day light application time is carry out culture of rootage under the environment of 12h, can obtain healthy and strong seedling of taking root after 25-30 days;
Step (6), hardening: step (5) is obtained take root seedling together with tissue culture bottle be placed in the rate of sheltering from heat or light be 80% green house in carry out in-bottle seeding 2 days, after unscrew bottle cap hardening 1 day again, then seedling removed medium and clean after medium, transplant in through pipeline high-temperature steam sterilization 1.8-2.2h with in the humus soil of the quality proportioning of 1:0.9-1.1 and perlite mixed-matrix, and build Small plastic shed film, after transplanting, air humidity remains on 70%-80%, by 2-3 week cellar culture, namely can be used as commercial seedling.
Generally need to reduce water spray frequency gradually, reduce rate of sheltering from heat or light and progressively open plastic film through 2-3 week cellar culture after transplanting of the present invention, rate of sheltering from heat or light finally can be reduced to 40%.
compared with prior art, its beneficial effect is in the present invention:
(1) acquisition time of explant of the present invention be March to October, can acquisition time long; In gatherer process, do not injure the subterraneous root be used as medicine, can continued growth after gathering, the plant also can not gathered with other to the harvest season is gathered in the crops simultaneously and does not affect Yield and qualities;
(2) group culturation rapid propagating technology of the present invention's employing, component is simple, easy and simple to handle, does not limit by region and season;
(3) fleabane flower tissue culture method of the present invention, reproduction speed is fast, reproduction rate is high.If calculated by a strain fleabane flower plant, a strain fleabane flower can gather 20-30 explant, within 40 days, can obtain callus and often for Multiplying culture 6-8 generation of 30 days through Fiber differentiation, then through the culture of rootage of 25 days and the hardening of 14-21 days.Calculate to breed for 6 generations, a strain fleabane flower seedling, is lasting less than in year, can obtain about 30-50 ten thousand commercial seedling.
Embodiment
Below in conjunction with embodiment, the present invention is described in further detail.
It will be understood to those of skill in the art that the following example only for illustration of the present invention, and should not be considered as limiting scope of the present invention.Unreceipted concrete technology or condition person in embodiment, according to the technology described by the document in this area or condition or carry out according to product description.Agents useful for same or the unreceipted production firm person of instrument, being can by buying the conventional products obtained.
Embodiment 1
A kind of fleabane flower tissue culture method, comprises the steps:
Step (1), the selection of explant: fleabane flower is colonizated in booth, management method waters the biological organic fertilizer liquid executed through abundant fermentation maturity routinely, chooses stalwartness, without damage by disease and insect plant, its young tender floral disc 100 of clip is as explant;
Step (2), the sterilization of explant: the explant will selected through step (1), rinse with liquid soap and wash 10min, remain to without liquid soap for 3 times with tap water, then explant is added to 2%(v/v) soak 8min in the Tween-80 aqueous solution, after using running water shower 1.9h again, be transferred on superclean bench and sterilize; During sterilization, first use 75%(v/v) alcohol disinfecting 10s, rear aseptic water washing 2 times, then with saturated aqueous sodium hypochlorite solution sterilization 10min, then use aseptic water washing 1 time, then use 0.05%(w/v) mercuric chloride solution sterilization 5min, finally use aseptic water washing 4 times;
Step (3), Fiber differentiation: after the explant disinfected through step (2) is cut off base portion wound, the inducing culture being seeded in MS+8.0mg/LIAA+5.0mg/L6-BA is cultivated, culture environment is temperature 23 DEG C, intensity of illumination 1500lux, odd-numbered day light application time is carry out Fiber differentiation under the condition of 12h, after 35 days, obtains the Cong Miao 100 pieces of band bud callus;
Step (4), Multiplying culture: the little Cong Cong Miaofen that step (3) obtains being cut to the 2-3 strain of band callus, then the proliferated culture medium being seeded in MS+0.5mg/LIAA+4.0mg/LKT is cultivated, culture environment is temperature 23 DEG C, intensity of illumination 1500lux, odd-numbered day light application time is carry out Multiplying culture under the condition of 12h, cultivates 30 days and after continuing switching 5 generation, obtains the tufted seedling 17500(100*5*7*5=17500 of callus) clump;
Step (5), culture of rootage: tufted seedling is divided and is cut to individual plant, then be seeded on the root media of 1/2MS+0.8mg/LIAA+2.0g/L active carbon, at 23 DEG C, intensity of illumination 1500lux, odd-numbered day light application time is carry out culture of rootage under the environment of 12h, within 25 days, obtains high about 5-8cm afterwards, the fleabane flower of tool 3-4 bar root takes root seedling about 120,000 strain;
Step (6), hardening: step (5) is obtained take root seedling together with tissue culture bottle be placed in the rate of sheltering from heat or light be 80% green house in carry out in-bottle seeding 2 days, after unscrew bottle cap hardening 1 day again, then seedling removed medium and clean after medium, by the planting density of 5cm*5cm plant in through pipeline high-temperature steam sterilization 1.8h with in the humus soil of the quality proportioning of 1:0.9 and perlite mixed-matrix, and build Small plastic shed film, after transplanting, air humidity remains on 70%-80%, by 2-3 week cellar culture, namely can be used as commercial seedling, after transplant in land for growing field crops, transplanting survival rate is 92.6%.
Embodiment 2
A kind of fleabane flower tissue culture method, comprises the steps:
Step (1), the selection of explant: fleabane flower is colonizated in booth, management method waters the biological organic fertilizer liquid executed through abundant fermentation maturity routinely, chooses stalwartness, without damage by disease and insect plant, its axillalry bud of clip (cut off blade and petiole, only leave petiole base) 100 is as explant;
Step (2), the sterilization of explant: the explant will selected through step (1), rinse with liquid soap and wash 20min, remain to without liquid soap for 4 times with tap water, then explant is added to 5%(v/v) soak 12min in the Tween-80 aqueous solution, after using running water shower 2.1h again, be transferred on superclean bench and sterilize; During sterilization, first use 75%(v/v) alcohol disinfecting 12s, rear aseptic water washing 1 time, again with saturated aqueous sodium hypochlorite solution sterilization 20min, then use aseptic water washing 2 times, then use 0.05%(w/v) mercuric chloride solution sterilization 10min, finally uses aseptic water washing 5 times;
Step (3), Fiber differentiation: after the explant disinfected through step (2) is cut off base portion wound, the inducing culture being seeded in MS+12.0mg/LIAA+8.0mg/L6-BA is cultivated, culture environment is temperature 25 DEG C, intensity of illumination 2000lux, odd-numbered day light application time is carry out Fiber differentiation under the condition of 12h, after 25 days, obtains the Cong Miao 100 pieces of band bud callus;
Step (4), Multiplying culture: the little Cong Cong Miaofen that step (3) obtains being cut to the 2-3 strain of band callus, then the proliferated culture medium being seeded in MS+1.0mg/LIAA+7.0mg/LKT is cultivated, culture environment is temperature 25 DEG C, intensity of illumination 2000lux, odd-numbered day light application time is carry out Multiplying culture under the condition of 12h, cultivates 25 days and after continuing switching 5 generation, obtains the tufted seedling 10000(100*4*5*5=10000 of band callus) clump;
Step (5), culture of rootage: tufted seedling is divided and is cut to individual plant, then be seeded on the root media of 1/2MS+1.2mg/LIAA+2.0g/L active carbon, at 25 DEG C, intensity of illumination 2000lux, odd-numbered day light application time is carry out culture of rootage under the environment of 12h, within 30 days, obtains high about 5-8cm afterwards, the fleabane flower of tool 3-4 bar root takes root seedling about 100,000 strain;
Step (6), hardening: step (5) is obtained take root seedling together with tissue culture bottle be placed in the rate of sheltering from heat or light be 80% green house in carry out in-bottle seeding 2 days, after unscrew bottle cap hardening 1 day again, then seedling removed medium and clean after medium, by the planting density of 5cm*5cm plant in through pipeline high-temperature steam sterilization 2.2h with in the humus soil of the quality proportioning of 1:1.1 and perlite mixed-matrix, and build Small plastic shed film, after transplanting, air humidity remains on 70%-80%, by 2-3 week cellar culture, namely can be used as commercial seedling, then transplant in land for growing field crops, transplanting survival rate is 90.7%.
Embodiment 3
A kind of fleabane flower tissue culture method, comprises the steps:
Step (1), the selection of explant: fleabane flower is colonizated in booth, management method waters the biological organic fertilizer liquid executed through abundant fermentation maturity routinely, chooses stalwartness, without damage by disease and insect plant, its young tender floral disc 100 of clip is as explant;
Step (2), the sterilization of explant: the explant will selected through step (1), rinse with liquid soap and wash 15min, remain to without liquid soap for 4 times with tap water, then explant is added to 2-5%(v/v) soak 10min in the Tween-80 aqueous solution, after using running water shower 2h again, be transferred on superclean bench and sterilize; During sterilization, first use 75%(v/v) alcohol disinfecting 11s, rear aseptic water washing 2 times, then with saturated aqueous sodium hypochlorite solution sterilization 13min, then use aseptic water washing 1 time, then use 0.05%(w/v) mercuric chloride solution sterilization 7min, finally use aseptic water washing 5 times;
Step (3), Fiber differentiation: after the explant disinfected through step (2) is cut off base portion wound, the inducing culture being seeded in MS+10.0mg/LIAA+7.0mg/L6-BA is cultivated, culture environment is temperature 24 DEG C, intensity of illumination 1800lux, odd-numbered day light application time is carry out Fiber differentiation under the condition of 12h, after 30 days, obtains band bud callus Cong Miao 100 pieces;
Step (4), Multiplying culture: the little Cong Cong Miaofen that step (3) obtains being cut to the 2-3 strain of band callus, then the proliferated culture medium being seeded in MS+0.8mg/LIAA+5.0mg/LKT is cultivated, culture environment is temperature 24 DEG C, intensity of illumination 1700lux, odd-numbered day light application time is carry out Multiplying culture under the condition of 12h, cultivates 35 days and after continuing switching 5 generation, obtains the Cong Miao 17500(100*5*7*5=17500 of band callus) clump;
Step (5), culture of rootage: tufted seedling is divided and is cut to individual plant, then be seeded on the root media of 1/2MS+1.1mg/LIAA+2.0g/L active carbon, at 24 DEG C, intensity of illumination 1900lux, odd-numbered day light application time is carry out culture of rootage under the environment of 12h, within 28 days, obtains high about 5-8cm afterwards, the fleabane flower of tool 3-4 bar root takes root seedling about 120,000 strain;
Step (6), hardening: step (5) is obtained take root seedling together with tissue culture bottle be placed in the rate of sheltering from heat or light be 80% green house in carry out in-bottle seeding 2 days, after unscrew bottle cap hardening 1 day again, then seedling removed medium and clean after medium, by the planting density of 5cm*5cm plant in through pipeline high-temperature steam sterilization 2h with in the humus soil of the quality proportioning of 1:1 and perlite mixed-matrix, and build Small plastic shed film, after transplanting, air humidity remains on 70%-80%, by 2-3 week cellar culture, namely can be used as commercial seedling, then transplant in land for growing field crops, transplanting survival rate is 91.7%.
More than show and describe general principle of the present invention, principal character and advantage of the present invention.The technical staff of the industry should understand; the present invention is not restricted to the described embodiments; what describe in above-described embodiment and specification just illustrates principle of the present invention; without departing from the spirit and scope of the present invention; the present invention also has various changes and modifications, and these changes and improvements all fall in the claimed scope of the invention.Application claims protection domain is defined by appending claims and equivalent thereof.
Claims (8)
1. a fleabane flower tissue culture method, is characterized in that, comprises the steps:
Step (1), the selection of explant: select the fleabane flower tender floral disc of children or axillalry bud as explant;
Step (2), the sterilization of explant: the explant will selected through step (1), clean totally with liquid soap and running water, be added to 2-5%(v/v) soak 8-12min in the Tween-80 aqueous solution, then running water shower 1.9-2.1h is used, finally successively with the sterilization of alcohol, clorox and mercuric chloride;
Step (3), Fiber differentiation: after the explant disinfected through step (2) is cut off base portion wound, the inducing culture being seeded in MS+8.0-12.0mg/LIAA+5.0-8.0mg/L6-BA is cultivated, culture environment is temperature 23-25 DEG C, intensity of illumination 1500-2000lux, odd-numbered day light application time is carry out Fiber differentiation under the condition of 12h, obtains Cong Miao;
Step (4), Multiplying culture: the little Cong Cong Miaofen that step (3) obtains being cut to the 2-3 strain of band callus, then the proliferated culture medium being seeded in MS+0.5-1.0mg/LIAA+4.0-7.0mg/LKT is cultivated, culture environment is temperature 23-25 DEG C, intensity of illumination 1500-2000lux, odd-numbered day light application time is carry out Multiplying culture under the condition of 12h, obtains tufted seedling;
Step (5), culture of rootage: divided by tufted seedling and be cut to individual plant, is then seeded on the root media of 1/2MS+0.8-1.2mg/LIAA+2.0g/L active carbon, at 23-25 DEG C, intensity of illumination 1500-2000lux, odd-numbered day light application time is carry out culture of rootage under the environment of 12h, obtains seedling of taking root;
Step (6), hardening: step (5) is obtained take root seedling together with tissue culture bottle be placed in the rate of sheltering from heat or light be 80% green house in carry out in-bottle seeding 2 days, after unscrew bottle cap hardening 1 day again, then seedling removed medium and clean after medium, transplant and plastic covering film, transplanting medium adopts humus soil: perlite=1:0.9-1.1, and after transplanting, air humidity remains on 70%-80%, by 2-3 week cellar culture, namely can be used as commercial seedling.
2. fleabane flower tissue culture method according to claim 1, it is characterized in that, the concrete grammar that step (2) middle liquid soap and running water clean is: rinsed by explant liquid soap after washing 10-20min, remains for 3-4 time with tap water to without liquid soap.
3. fleabane flower tissue culture method according to claim 1, it is characterized in that, in step (2), the concrete grammar of sterilization is: first use 75%(v/v) alcohol disinfecting 10-12s, after use aseptic water washing 1-2 time, again with saturated aqueous sodium hypochlorite solution sterilization 10-20min, then use aseptic water washing 1-2 time, then use 0.05%(w/v) mercuric chloride solution sterilization 5-10min, finally uses aseptic water washing 4-5 time.
4. fleabane flower tissue culture method according to claim 1, is characterized in that, in step (2), sterilization is carried out on superclean bench.
5. fleabane flower tissue culture method according to claim 1, is characterized in that, the time of the Fiber differentiation described in step (3) is 25-35 days.
6. fleabane flower tissue culture method according to claim 1, is characterized in that, the subculture cycle of the Multiplying culture described in step (4) is 25-35 days.
7. fleabane flower tissue culture method according to claim 1, is characterized in that, the incubation time of the culture of rootage described in step (5) is 25-30 days.
8. fleabane flower tissue culture method according to claim 1, is characterized in that, comprise the steps:
Step (1), the selection of explant: fleabane flower is colonizated in booth, management method waters the biological organic fertilizer liquid executed through abundant fermentation maturity routinely, chooses stalwartness, without damage by disease and insect plant, its young tender floral disc of clip or axillalry bud are as explant;
Step (2), the sterilization of explant: the explant will selected through step (1), rinse with liquid soap and wash 10-20min, remain to without liquid soap for 3-4 time with tap water, then explant is added to 2-5%(v/v) soak 8-12min in the Tween-80 aqueous solution, after using running water shower 2h again, be transferred on superclean bench and sterilize; During sterilization, first use 75%(v/v) alcohol disinfecting 10-12s, rear aseptic water washing 1-2 time, again with saturated aqueous sodium hypochlorite solution sterilization 10-20min, then use aseptic water washing 1-2 time, then use 0.05%(w/v) mercuric chloride solution sterilization 5-10min, finally uses aseptic water washing 4-5 time;
Step (3), Fiber differentiation: after the explant disinfected through step (2) is cut off base portion wound, the inducing culture being seeded in MS+8.0-12.0mg/LIAA+5.0-8.0mg/L6-BA is cultivated, culture environment is temperature 23-25 DEG C, intensity of illumination 1500-2000lux, odd-numbered day light application time is carry out Fiber differentiation under the condition of 12h, and after 25-35 days, an one-step inducing goes out the Cong Miao with callus;
Step (4), Multiplying culture: the little Cong Cong Miaofen that step (3) obtains being cut to the 2-3 strain of band callus, then the proliferated culture medium being seeded in MS+0.5-1.0mg/LIAA+4.0-7.0mg/LKT is cultivated, culture environment is temperature 23-25 DEG C, intensity of illumination 1500-2000lux, odd-numbered day light application time is carry out Multiplying culture under the condition of 12h, within 25-35 days, is a subculture cycle, obtains tufted seedling;
Step (5), culture of rootage: tufted seedling is divided and is cut to individual plant, then be seeded on the root media of 1/2MS+0.8-1.2mg/LIAA+2.0g/L active carbon, at 23-25 DEG C, intensity of illumination 1500-2000lux, odd-numbered day light application time is carry out culture of rootage under the environment of 12h, can obtain healthy and strong seedling of taking root after 25-30 days;
Step (6), hardening: step (5) is obtained take root seedling together with tissue culture bottle be placed in the rate of sheltering from heat or light be 80% green house in carry out in-bottle seeding 2 days, after unscrew bottle cap hardening 1 day again, then seedling removed medium and clean after medium, plant in through pipeline high-temperature steam sterilization 1.8-2.2h with in the humus soil of the quality proportioning of 1:0.9-1.1 and perlite mixed-matrix, and build Small plastic shed film, after transplanting, air humidity remains on 70%-80%, by 2-3 week cellar culture, namely can be used as commercial seedling.
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Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN106069752A (en) * | 2016-06-15 | 2016-11-09 | 上海市农业科学院 | Taking root and hardening off method of a kind of African Chrysanthemum plantlet in vitro |
| CN107047201A (en) * | 2017-05-10 | 2017-08-18 | 蚌埠市金牛湾农业科技发展有限公司 | A kind of seed seedling-raising method of ligustrum lucidum ait |
| CN117158319A (en) * | 2023-09-19 | 2023-12-05 | 云南农业大学 | Erigeron breviscapus plant tissue culture and plant regeneration method |
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| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN106069752A (en) * | 2016-06-15 | 2016-11-09 | 上海市农业科学院 | Taking root and hardening off method of a kind of African Chrysanthemum plantlet in vitro |
| CN106069752B (en) * | 2016-06-15 | 2018-05-08 | 上海市农业科学院 | A kind of African Chrysanthemum tissue-cultured seedling take root and hardening off method |
| CN107047201A (en) * | 2017-05-10 | 2017-08-18 | 蚌埠市金牛湾农业科技发展有限公司 | A kind of seed seedling-raising method of ligustrum lucidum ait |
| CN117158319A (en) * | 2023-09-19 | 2023-12-05 | 云南农业大学 | Erigeron breviscapus plant tissue culture and plant regeneration method |
| CN117158319B (en) * | 2023-09-19 | 2024-04-23 | 云南农业大学 | Erigeron breviscapus plant tissue culture and plant regeneration method |
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| CN105075860B (en) | 2017-09-19 |
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