CN104304037B - A kind of Dendrobium fimbriatum Hook. seed tissue cultivates method for quickly breeding - Google Patents
A kind of Dendrobium fimbriatum Hook. seed tissue cultivates method for quickly breeding Download PDFInfo
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Abstract
The invention discloses a kind of Dendrobium fimbriatum Hook. seed tissue and cultivate method for quickly breeding, by the planting seed of Dendrobium fimbriatum Hook. sprout-induction in inducing culture, then turn Dendrobium fimbriatum Hook. budlet successively to move to and subculture medium, root media continue cultivate, obtain Dendrobium fimbriatum Hook. to take root tissue cultured seedling, can transplant to booth, carry out seedling exercising.The method for tissue culture of the present invention is easy, and step is simple, easy to operate, and gained Dendrobium fimbriatum Hook. is taken root tissue cultured seedling robust plant, and energy for growth is strong, is suitable to factorial praluction.
Description
Technical field
The present invention relates to rapid breeding method of plant, particularly a kind of Dendrobium fimbriatum Hook. seed tissue cultivates method for quickly breeding.
Background technology
Herba Dendrobii is the fresh or dry stem of orchid Dendrobium nobile DendrobiumnobileLindl., Herba Dendrobii DendrobiumcandidumWall.exLindl. or Herba Dendrobii oculati DendrobiumfimbriatumHook.var.oculatumHook. and allied species thereof.In China's traditional medicine, Herba Dendrobii is conventional valuable ingredient of Chinese medicine, begins to be loaded in Shennong's Herbal, is classified as top grade.
nullDendrobium fimbriatum Hook. (DendrobiumfimbriatumHook.var.oculatumHook.) is the orchid family Dendrobium Sw，Stem is sturdy，Tiltedly vertical or sagging，Quality is hard，Slightly in spindle above cylinder or sometimes base portion，Ye Erlie，Keratin，Long Circle or oval shape lanceolar，Raceme length 5～15 centimetres，Dredge raw 6～12 flowers，Spend golden yellow，Quality is thin，Carry out，Slightly has fragrance，4～June of florescence，Produce south Guangxi to (Tiane, the northwestward、Reach the clouds、Tianlin County、Longzhou、It etc.、Long Lin、Donglan、Wuming、Jingxi、Nandan)、South of Guizhou is to (Luodian, the west and south、Xingyi、Only mountain)、Southeastern Yunnan is to (Xichou, the west and south、Mengzi、Stone screen、Rich people、Simao、Menhai、Cang Yuan、Zhenkang)，Height above sea level 600～1700 meters，It is born in thick forest on the dark and damp rock on trunk or mountain valley，It is distributed in India、Nepal、Sillim、Bhutan、Burma、Thailand、Vietnam." Chinese Pharmacopoeia " claims Herba Dendrobii oculati, has reinforcing stomach reg fluid, promoting the production of body fluid to quench thirst.Losing for cloudy impairment of body fluid, xerostomia excessive thirst, lack of appetite is retched, deficiency-heat after being ill, poor vision.Modern study shows, the EN that finds in Dendrobium fimbriatum Hook., hair orchid phenanthrene, the chemical composition such as chrysotoxene have antitumor action.
It is difficult in the wild due to Orchid Seeds germinate, natural propagation is mainly by division propagation, breeding potential underground, natural renovation difficulty, development along with society, the people, to beautiful pursuit and demand medicinal for a long time, cause the heavy damage of wild resource, and the Dendrobium fimbriatum Hook. of beauty seldom can have been seen in existing field.
Summary of the invention
It is an object of the invention to provide a kind of Dendrobium fimbriatum Hook. seed tissue and cultivate method for quickly breeding, the seed directly taking Dendrobium fimbriatum Hook. carries out tissue culture, effectively shorten the cultivation cycle of Dendrobium fimbriatum Hook., reduce the mutation probability because repeatedly successive transfer culture causes, adopting the Dendrobium fimbriatum Hook. tissue cultured seedling of tissue culture's gained, robust plant, energy for growth is strong, be suitable to factorial praluction, to solve the seedling problem of Dendrobium fimbriatum Hook. artificial growth.
Present invention provide the technical scheme that
A kind of Dendrobium fimbriatum Hook. seed tissue cultivates method for quickly breeding, takes Dendrobium fimbriatum Hook. seed and carries out in-vitro inducing cultivation as material, obtains the tissue culturing seedling of Dendrobium fimbriatum Hook..
Preferably, described Dendrobium fimbriatum Hook. seed tissue is cultivated in method for quickly breeding, comprises the following steps:
Step one, by the Dendrobium fimbriatum Hook. planting seed of pretreatment on inducing culture, cultivate 7-15 days, obtain Dendrobium fimbriatum Hook. budlet, wherein, described inducing culture is: with MS-H for minimal medium, add 0.5-3.0mg/L heteroauxing, 1.0-5.0mg/L naphthalene acetic acid, 0.1-1.0mg/L Thidiazuron, 25-40g/L sucrose and 3.0-5.0g/L agar, and the pH value adjusting inducing culture is 5.6-6.0；
Step 2, Dendrobium fimbriatum Hook. budlet is placed in subculture medium, cultivate 30-40 days, obtain Dendrobium fimbriatum Hook. group training differentiation Seedling, wherein, described subculture medium is: with MS for minimal medium, add 0.2.0-2.0mg/L kinetins, 2.0-4.0mg/L6-benayl aminopurine, 1.0-4.0mg/L naphthalene acetic acid, 0.5-3.0mg/L indolepopionic acid, 0.1-2.0mg/L Thidiazuron, 25-40g/L sucrose and 3.0-5.0g/L agar, and the pH value adjusting subculture medium is 5.6-6.0；
Step 3, by the training differentiation transplantation of seedlings of Dendrobium fimbriatum Hook. group in root media, after cultivating 30-40 days, being transferred under nature light and irradiate cultivation 40-50 days, it is thus achieved that Dendrobium fimbriatum Hook. is taken root tissue cultured seedling；Wherein, described root media is: with 1/2MS for minimal medium, adding 5-20% Fructus Musae, 0.1-4.0mg/L naphthalene acetic acid, 2.0-4.0mg/L6-benayl aminopurine, 2-5% sucrose and 0.2-0.5% activated carbon, and the pH value adjusting root media is 5.8-6.2；
Step 4, tissue cultured seedling of being taken root by Dendrobium fimbriatum Hook. are transplanted to seedbed, treat that the growth of Dendrobium fimbriatum Hook. seedling is stable, can transplant to booth.
Preferably, described Dendrobium fimbriatum Hook. seed tissue is cultivated in method for quickly breeding, the pretreatment of Dendrobium fimbriatum Hook. seed described in step one is, first by Dendrobium fimbriatum Hook. outstanding achievement with the 80-85% alcohol solution dipping 20-40 second, mercuric chloride is sterilized 10-20 minute, cut the outstanding achievement of Dendrobium fimbriatum Hook. after cleaning up, obtain the Dendrobium fimbriatum Hook. seed of pretreatment.
Preferably, described Dendrobium fimbriatum Hook. seed tissue is cultivated in method for quickly breeding, and the medium of seedling bed described in step 4 is: 20-30 bark, 10-20 weedtree, 20-30 Vermiculitum and 10-15 perlite.
Preferably, described Dendrobium fimbriatum Hook. seed tissue is cultivated in method for quickly breeding, and tissue cultured seedling of being taken root by Dendrobium fimbriatum Hook. in step 4 is transplanted to before seedbed, and the culture medium of tissue cultured seedling root of first being taken root by Dendrobium fimbriatum Hook. cleans up, and dries steam, is transplanting to seedbed.
Preferably, described Dendrobium fimbriatum Hook. seed tissue is cultivated in method for quickly breeding, and described step one is to step 3, and Dendrobium fimbriatum Hook. condition of culture in culture medium is: cultivation temperature is 28-32 DEG C, intensity of illumination is 2500-3000lux, and light application time is 13-15h/d.
The invention has the beneficial effects as follows: the present invention adopts modern biotechnology, clone Dendrobium fimbriatum Hook., large-scale production seedling, solve the seedling problem of Dendrobium fimbriatum Hook. artificial growth, contribute for the development of society and the protection of Diffuse Coptosapelta wild plant resource.
In the present invention, Dendrobium fimbriatum Hook. seed tissue cultivates method for quickly breeding, the germination percentage of Seed inducement reaches 96%, planting percent 95% after successive transfer culture, it is transferred in root media, the rooting rate of Dendrobium fimbriatum Hook. is 90%, the survival rate finally moving into booth seedling exercising is 94%, substantially increases tissue culture's efficiency of Dendrobium fimbriatum Hook.；Although tender shoots and seed can serve as the original material of group training, but Herba Dendrobii tender shoots is the portion of material taken on plant from the outside, due to a strain Herba Dendrobii only one of which bud, also several bud can only be obtained by stem section, each bud adopt after sterilization, the sterilizable material finally obtained probably only has about 40%, then sterilizable material is utilized could to continue propagation, induction protocorm, expanding propagation, will through the very long time, it is generally required to abundant material could be obtained through 10-20 generation, but group training process relaying number from generation to generation is more long, cost raises, vitrification variation strengthens, therefore material now has not had very big value, increase along with subculture, material degeneration is also very serious problem.The seed using Dendrobium fimbriatum Hook. in the present invention is cultivated, germination is significantly shorter than tender shoots subculture to required time of finally emerging, it it is almost the odd of tender shoots, this has saved fund to a great extent, save the energy, improve competitiveness, only within more than 200 day, can emerge needed for utilizing seed, and utilizing tender shoots required time of accomplishing to emerge will be more than 1000 day, therefore consider also to be far superior to tender shoots induction Seedling from market angle.Relative tender shoots tissue culture, the cost of material is low as group training for the seed of Dendrobium fimbriatum Hook. of utilizing of the present invention, promotes easily; good economy performance, advanced technology, and there is significantly high market value; interest concessions plantation, is conducive to expanding and promotes, the protection of Dendrobium Plants is provided new thinking.
Detailed description of the invention
The present invention is described in further detail below, to make those skilled in the art can implement according to this with reference to description word.
A kind of Dendrobium fimbriatum Hook. seed tissue cultivates method for quickly breeding, takes Dendrobium fimbriatum Hook. seed and carries out in-vitro inducing cultivation as material, obtains the tissue culturing seedling of Dendrobium fimbriatum Hook..
Described Dendrobium fimbriatum Hook. seed tissue is cultivated in method for quickly breeding, comprises the following steps:
nullStep one、First by Dendrobium fimbriatum Hook. outstanding achievement with the 80-85% alcohol solution dipping 20-40 second，Mercuric chloride is sterilized 10-20 minute，The outstanding achievement of Dendrobium fimbriatum Hook. is cut after cleaning up，Obtain the Dendrobium fimbriatum Hook. seed of pretreatment，By the Dendrobium fimbriatum Hook. planting seed of pretreatment on inducing culture，It is 28-32 DEG C in cultivation temperature，Intensity of illumination is 2500-3000lux，Cultivate 7-15 days when light application time is 13-15h/d，Obtain Dendrobium fimbriatum Hook. budlet，Wherein，Described inducing culture is: with MS-H for minimal medium，Add 0.5-3.0mg/L heteroauxing、1.0-5.0mg/L naphthalene acetic acid、0.1-1.0mg/L Thidiazuron、25-40g/L sucrose and 3.0-5.0g/L agar，And the pH value adjusting inducing culture is 5.6-6.0；Wherein, in inducing culture, heteroauxing is ubiquitous Endogenous auxin in a kind of plant, promotes Dendrobium fimbriatum Hook. seed quick-speed germination budlet；Sucrose is energy substance, provides carbon source and the energy for plant tissue culture, moreover it is possible to well maintains the hypotonic environment in culture medium, can also reduce the pollution of microorganism simultaneously to a certain extent, improves the gnotobasis quality of Dendrobium fimbriatum Hook.；The inducing culture adopted substantially increases the Calli Differentiation ability of Dendrobium fimbriatum Hook. so that the bud ratio of Dendrobium fimbriatum Hook. budlet is up to 92%.
Step 2, Dendrobium fimbriatum Hook. budlet is placed in subculture medium, it is 28-32 DEG C in cultivation temperature, intensity of illumination is 2500-3000lux, cultivate 30-40 days when light application time is 13-15h/d, obtain Dendrobium fimbriatum Hook. group training differentiation Seedling highly ordered for 2-4cm, wherein, described subculture medium is: with MS for minimal medium, add 0.2.0-2.0mg/L kinetins, 2.0-4.0mg/L6-benayl aminopurine, 1.0-4.0mg/L naphthalene acetic acid, 0.5-3.0mg/L indolepopionic acid, 0.1-2.0mg/L Thidiazuron, 25-40g/L sucrose and 3.0-5.0g/L agar, and the pH value adjusting subculture medium is 5.6-6.0；Wherein, in subculture medium, kinetins is 6-furfuryl group adenine, is phytocytomine, effectively facilitates the cell division of Dendrobium fimbriatum Hook.；6-benzyl aminopurine effectively accelerates plant cell growth, promotes growth and development of plants, the differentiation of induction Dendrobium fimbriatum Hook. budlet, promotes sprouting of lateral bud growth；Naphthalene acetic acid is broad spectrum type plant growth regulator, can effectively facilitate cell division and expansion, induced synthesis adventitious root；Sucrose is energy substance, provides carbon source and the energy for plant tissue culture, moreover it is possible to well maintain the hypotonic environment in culture medium；The planting percent of Dendrobium fimbriatum Hook. group training differentiation Seedling is 98%.
Step 3, by the training differentiation transplantation of seedlings of Dendrobium fimbriatum Hook. group in root media, it is 28-32 DEG C in cultivation temperature, intensity of illumination is 2500-3000lux, after cultivating 30-40 days when light application time is 13-15h/d, it is transferred under nature light to irradiate and cultivates 40-50 days, it is thus achieved that Dendrobium fimbriatum Hook. is taken root tissue cultured seedling；Wherein, described root media is: with 1/2MS for minimal medium, adding 5-20% Fructus Musae, 0.1-4.0mg/L naphthalene acetic acid, 2.0-4.0mg/L6-benayl aminopurine, 2-5% sucrose and 0.2-0.5% activated carbon, and the pH value adjusting root media is 5.8-6.2；Wherein, root media adds the Fructus Musae smashed, promote Dendrobium fimbriatum Hook. group training differentiation Seedling Rapid Rooting, desired nutritional material is provided for Dendrobium fimbriatum Hook. group training differentiation Seedling, naphthalene acetic acid is broad spectrum type plant growth regulator, cell division and expansion can be effectively facilitated, induced synthesis adventitious root, sucrose is energy substance, thering is provided energy, being added with of activated carbon helps adsorb some harmful metabolites, and activated carbon makes root media blackening simultaneously, dark condition can be imitated, be conducive to taking root of group training differentiation Seedling；It is placed under nature light and irradiates after Dendrobium fimbriatum Hook. group training differentiation seedling rooting, provide the culture environment similar to outdoor environment for group training differentiation Seedling, improve Dendrobium fimbriatum Hook. group training differentiation Seedling survival rate in later stage seedling exercising process.
Step 4, tissue cultured seedling root that Dendrobium fimbriatum Hook. is taken root culture medium with cleaning up, dry steam, tissue cultured seedling of being taken root by Dendrobium fimbriatum Hook. again is transplanted to seedbed, the substrate wherein arranged on seedbed is: 20-30 bark, 10-20 weedtree, 20-30 Vermiculitum and 10-15 perlite, cultivate 30-40 days, Dendrobium fimbriatum Hook. is taken root after tissue cultured seedling growth is stablized and can be transplanted to booth, suitably to shelter from heat or light during greenhouse cultivation, controlling shade density is 75%, water in interval, temperature is 28 DEG C, wants cascade to lower the temperature more than 28 DEG C, it is prevented that root rot occurs.
Embodiment 1
A kind of Dendrobium fimbriatum Hook. seed tissue cultivates method for quickly breeding, takes Dendrobium fimbriatum Hook. seed and carries out in-vitro inducing cultivation as material, obtains the tissue culturing seedling of Dendrobium fimbriatum Hook..
Described Dendrobium fimbriatum Hook. seed tissue is cultivated in method for quickly breeding, comprises the following steps:
Step one, first by Dendrobium fimbriatum Hook. outstanding achievement 80% alcohol solution dipping 20 seconds, mercuric chloride is sterilized 10 minutes, the outstanding achievement of Dendrobium fimbriatum Hook. is cut after cleaning up, obtain the Dendrobium fimbriatum Hook. seed of pretreatment, by the Dendrobium fimbriatum Hook. planting seed of pretreatment on inducing culture, it it is 28 DEG C in cultivation temperature, intensity of illumination is 2600lux, cultivate 10 days when light application time is 15h/d, obtain Dendrobium fimbriatum Hook. budlet, wherein, described inducing culture is: with MS-H for minimal medium, add 2.0mg/L heteroauxing, 2.0mg/L naphthalene acetic acid, 0.5mg/L Thidiazuron, 25g/L sucrose and 3.0g/L agar, and the pH value adjusting inducing culture is 5.9；
Step 2, Dendrobium fimbriatum Hook. budlet is placed in subculture medium, it it is 28 DEG C in cultivation temperature, intensity of illumination is 2600lux, cultivate 40 days when light application time is 15h/d, obtain Dendrobium fimbriatum Hook. group training differentiation Seedling highly ordered for 3cm, wherein, described subculture medium is: with MS for minimal medium, add 0.5mg/L kinetins, 2.0mg/L6-benayl aminopurine, 1.0mg/L naphthalene acetic acid, 2.0mg/L indolepopionic acid, 0.5mg/L Thidiazuron, 40g/L sucrose and 4.0g/L agar, and the pH value adjusting subculture medium is 5.9；
Step 3, by the training differentiation transplantation of seedlings of Dendrobium fimbriatum Hook. group in root media, being 28 DEG C in cultivation temperature, intensity of illumination is 2600lux, after cultivating 35 days when light application time is 15h/d, it is transferred under nature light to irradiate and cultivates 50 days, it is thus achieved that Dendrobium fimbriatum Hook. is taken root tissue cultured seedling；Wherein, described root media is: with 1/2MS for minimal medium, and adding 10% Fructus Musae, 1.0mg/L naphthalene acetic acid, 2.0mg/L6-benayl aminopurine, 3% sucrose and 0.2% activated carbon, and the pH value adjusting root media is 5.9；
Step 4, tissue cultured seedling root that Dendrobium fimbriatum Hook. is taken root culture medium with cleaning up, dry steam, tissue cultured seedling of being taken root by Dendrobium fimbriatum Hook. again is transplanted to seedbed, the substrate wherein arranged on seedbed is: 20g bark, 15g weedtree, 25g Vermiculitum and 15g perlite, cultivate 40 days, Dendrobium fimbriatum Hook. is taken root after tissue cultured seedling growth is stablized and can be transplanted to booth, suitably to shelter from heat or light during greenhouse cultivation, controlling shade density is 75%, water in interval, temperature is 28 DEG C, wants cascade to lower the temperature more than 28 DEG C, it is prevented that root rot occurs.
Embodiment 2
A kind of Dendrobium fimbriatum Hook. seed tissue cultivates method for quickly breeding, takes Dendrobium fimbriatum Hook. seed and carries out in-vitro inducing cultivation as material, obtains the tissue culturing seedling of Dendrobium fimbriatum Hook..
Described Dendrobium fimbriatum Hook. seed tissue is cultivated in method for quickly breeding, comprises the following steps:
Step one, first by Dendrobium fimbriatum Hook. outstanding achievement 80% alcohol solution dipping 30 seconds, mercuric chloride is sterilized 20 minutes, the outstanding achievement of Dendrobium fimbriatum Hook. is cut after cleaning up, obtain the Dendrobium fimbriatum Hook. seed of pretreatment, by the Dendrobium fimbriatum Hook. planting seed of pretreatment on inducing culture, it it is 30 DEG C in cultivation temperature, intensity of illumination is 2800lux, cultivate 15 days when light application time is 14h/d, obtain Dendrobium fimbriatum Hook. budlet, wherein, described inducing culture is: with MS-H for minimal medium, add 1.0mg/L heteroauxing, 3.0mg/L naphthalene acetic acid, 0.8mg/L Thidiazuron, 30g/L sucrose and 4.0g/L agar, and the pH value adjusting inducing culture is 6.0；
Step 2, Dendrobium fimbriatum Hook. budlet is placed in subculture medium, it it is 30 DEG C in cultivation temperature, intensity of illumination is 2800lux, cultivate 30 days when light application time is 14h/d, obtain Dendrobium fimbriatum Hook. group training differentiation Seedling highly ordered for 2-4cm, wherein, described subculture medium is: with MS for minimal medium, add 2.0mg/L kinetins, 4.0mg/L6-benayl aminopurine, 3.0mg/L naphthalene acetic acid, 2.0mg/L indolepopionic acid, 1.0mg/L Thidiazuron, 30g/L sucrose and 4.0g/L agar, and the pH value adjusting subculture medium is 6.0；
Step 3, by the training differentiation transplantation of seedlings of Dendrobium fimbriatum Hook. group in root media, being 30 DEG C in cultivation temperature, intensity of illumination is 2800lux, after cultivating 35 days when light application time is 14h/d, it is transferred under nature light to irradiate and cultivates 45 days, it is thus achieved that Dendrobium fimbriatum Hook. is taken root tissue cultured seedling；Wherein, described root media is: with 1/2MS for minimal medium, and adding 15% Fructus Musae, 2.0mg/L naphthalene acetic acid, 3.0mg/L6-benayl aminopurine, 2% sucrose and 0.5% activated carbon, and the pH value adjusting root media is 6.0；
Step 4, tissue cultured seedling root that Dendrobium fimbriatum Hook. is taken root culture medium with cleaning up, dry steam, tissue cultured seedling of being taken root by Dendrobium fimbriatum Hook. again is transplanted to seedbed, the substrate wherein arranged on seedbed is: 30g bark, 15g weedtree, 20g Vermiculitum and 10g perlite, cultivate 30 days, Dendrobium fimbriatum Hook. is taken root after tissue cultured seedling growth is stablized and can be transplanted to booth, suitably to shelter from heat or light during greenhouse cultivation, controlling shade density is 75%, water in interval, temperature is 28 DEG C, wants cascade to lower the temperature more than 28 DEG C, it is prevented that root rot occurs.
Embodiment 3
A kind of Dendrobium fimbriatum Hook. seed tissue cultivates method for quickly breeding, takes Dendrobium fimbriatum Hook. seed and carries out in-vitro inducing cultivation as material, obtains the tissue culturing seedling of Dendrobium fimbriatum Hook..
Described Dendrobium fimbriatum Hook. seed tissue is cultivated in method for quickly breeding, comprises the following steps:
Step one, first by Dendrobium fimbriatum Hook. outstanding achievement 85% alcohol solution dipping 40 seconds, mercuric chloride is sterilized 20 minutes, the outstanding achievement of Dendrobium fimbriatum Hook. is cut after cleaning up, obtain the Dendrobium fimbriatum Hook. seed of pretreatment, by the Dendrobium fimbriatum Hook. planting seed of pretreatment on inducing culture, it it is 32 DEG C in cultivation temperature, intensity of illumination is 3000lux, cultivate 14 days when light application time is 13h/d, obtain Dendrobium fimbriatum Hook. budlet, wherein, described inducing culture is: with MS-H for minimal medium, add 3.0mg/L heteroauxing, 4.0mg/L naphthalene acetic acid, 1.0mg/L Thidiazuron, 40g/L sucrose and 5.0g/L agar, and the pH value adjusting inducing culture is 6.0；
Step 2, Dendrobium fimbriatum Hook. budlet is placed in subculture medium, it it is 32 DEG C in cultivation temperature, intensity of illumination is 3000lux, cultivate 38 days when light application time is 13h/d, obtain Dendrobium fimbriatum Hook. group training differentiation Seedling highly ordered for 2-4cm, wherein, described subculture medium is: with MS for minimal medium, add 1.0mg/L kinetins, 2.0mg/L6-benayl aminopurine, 1.0mg/L naphthalene acetic acid, 1.0mg/L indolepopionic acid, 0.5mg/L Thidiazuron, 25g/L sucrose and 3.0g/L agar, and the pH value adjusting subculture medium is 6.0；
Step 3, by the training differentiation transplantation of seedlings of Dendrobium fimbriatum Hook. group in root media, being 32 DEG C in cultivation temperature, intensity of illumination is 3000lux, after cultivating 30 days when light application time is 13h/d, it is transferred under nature light to irradiate and cultivates 40 days, it is thus achieved that Dendrobium fimbriatum Hook. is taken root tissue cultured seedling；Wherein, described root media is: with 1/2MS for minimal medium, and adding 20% Fructus Musae, 3.0mg/L naphthalene acetic acid, 4.0mg/L6-benayl aminopurine, 2% sucrose and 0.2% activated carbon, and the pH value adjusting root media is 6.0；
Step 4, tissue cultured seedling root that Dendrobium fimbriatum Hook. is taken root culture medium with cleaning up, dry steam, tissue cultured seedling of being taken root by Dendrobium fimbriatum Hook. again is transplanted to seedbed, the substrate wherein arranged on seedbed is: 20g bark, 15g weedtree, 20g Vermiculitum and 10g perlite, cultivate 40 days, Dendrobium fimbriatum Hook. is taken root after tissue cultured seedling growth is stablized and can be transplanted to booth, suitably to shelter from heat or light during greenhouse cultivation, controlling shade density is 75%, water in interval, temperature is 28 DEG C, wants cascade to lower the temperature more than 28 DEG C, it is prevented that root rot occurs.
Test is compared
(1) group 1 is that Dendrobium fimbriatum Hook. seed tissue of the present invention cultivates method for quickly breeding, adopts Dendrobium fimbriatum Hook. seed as group training material；Culture medium is respectively as follows:
Inducing culture, with MS-H for minimal medium, adds 0.5-3.0mg/L heteroauxing, 1.0-5.0mg/L naphthalene acetic acid, 0.1-1.0mg/L Thidiazuron, 25-40g/L sucrose and 3.0-5.0g/L agar；
Subculture medium is: with MS for minimal medium, adds 0.2.0-2.0mg/L kinetins, 2.0-4.0mg/L6-benayl aminopurine, 1.0-4.0mg/L naphthalene acetic acid, 0.5-3.0mg/L indolepopionic acid, 0.1-2.0mg/L Thidiazuron, 25-40g/L sucrose and 3.0-5.0g/L agar；
Root media is: with 1/2MS for minimal medium, adds 5-20% Fructus Musae, 0.1-4.0mg/L naphthalene acetic acid, 2.0-4.0mg/L6-benayl aminopurine, 2-5% sucrose and 0.2-0.5% activated carbon.
(2) group 2 is for adopting Dendrobium fimbriatum Hook. tender shoots as group training material, and the culture medium of employing is identical with group 1；
(3) group 3 is for adopting Dendrobium fimbriatum Hook. seed as group training material, and the culture medium of employing is respectively as follows: compared to group 1
Inducing culture is MS-H is minimal medium, adds 0.5-3.0mg/L heteroauxing, 1.0-5.0mg/L naphthalene acetic acid, 25-40g/L sucrose and 3.0-5.0g/L agar；
Subculture medium is with MS for minimal medium, with MS for minimal medium, add 0.2.0-2.0mg/L kinetins, 2.0-4.0mg/L6-benayl aminopurine, 1.0-4.0mg/L naphthalene acetic acid, 0.5-3.0mg/L indolepopionic acid, 25-40g/L sucrose and 3.0-5.0g/L agar；
Root media: add 5-20% Fructus Musae, 0.1-4.0mg/L naphthalene acetic acid, 2.0-4.0mg/L6-benayl aminopurine, 2-5% sucrose and 0.2-0.5% activated carbon.
(4) group 4 is for adopting Dendrobium fimbriatum Hook. seed as group training material, and the culture medium of employing is respectively as follows: compared to group 1
Inducing culture, with MS-H for minimal medium, adds 1.0-5.0mg/L naphthalene acetic acid, 0.1-1.0mg/L Thidiazuron, 25-40g/L sucrose and 3.0-5.0g/L agar；
Subculture medium is: with MS for minimal medium, adds 0.2.0-2.0mg/L kinetins, 2.0-4.0mg/L6-benayl aminopurine, 1.0-4.0mg/L naphthalene acetic acid, 0.1-2.0mg/L Thidiazuron, 25-40g/L sucrose and 3.0-5.0g/L agar；
Root media is: with 1/2MS for minimal medium, adds 5-20% Fructus Musae, 0.1-4.0mg/L naphthalene acetic acid, 2.0-4.0mg/L6-benayl aminopurine, 2-5% sucrose and 0.2-0.5% activated carbon.
(5) group 5 is for adopting Dendrobium fimbriatum Hook. seed as group training material, and the culture medium of employing is respectively as follows: compared to group 1
Inducing culture, with MS-H for minimal medium, adds 0.5-3.0mg/L heteroauxing, 0.1-1.0mg/L Thidiazuron, 25-40g/L sucrose and 3.0-5.0g/L agar；
Subculture medium is: with MS for minimal medium, adds 0.2.0-2.0mg/L kinetins, 2.0-4.0mg/L6-benayl aminopurine, 0.5-3.0mg/L indolepopionic acid, 0.1-2.0mg/L Thidiazuron, 25-40g/L sucrose and 3.0-5.0g/L agar；
Root media is: with 1/2MS for minimal medium, adds 5-20% Fructus Musae, 2.0-4.0mg/L6-benayl aminopurine, 2-5% sucrose and 0.2-0.5% activated carbon.
(6) group 6 is for adopting Dendrobium fimbriatum Hook. seed as group training material, and the culture medium of employing is respectively as follows: compared to group 1
Inducing culture, with MS-H for minimal medium, adds 0.5-3.0mg/L heteroauxing, 1.0-5.0mg/L naphthalene acetic acid, 0.1-1.0mg/L Thidiazuron, 25-40g/L sucrose and 3.0-5.0g/L agar；
Subculture medium is: with MS for minimal medium, adds 1.0-4.0mg/L naphthalene acetic acid, 0.5-3.0mg/L indolepopionic acid, 0.1-2.0mg/L Thidiazuron, 25-40g/L sucrose and 3.0-5.0g/L agar；
Root media is: with 1/2MS for minimal medium, adds 5-20% Fructus Musae, 0.1-4.0mg/L naphthalene acetic acid, 2-5% sucrose and 0.2-0.5% activated carbon.
(7) group 7 is for adopting Dendrobium fimbriatum Hook. seed as group training material, and the culture medium of employing is respectively as follows: compared to group 1
Inducing culture with MS-H for culture medium, 25-40g/L sucrose and 3.0-5.0g/L agar；
Subculture medium is: with MS for minimal medium, 25-40g/L sucrose and 3.0-5.0g/L agar；
Root media is: with 1/2MS for minimal medium, 2-5% sucrose.
Obtain result as shown in table 1:
Table 1
As can be seen from Table 1, group 1 compares with group 2, the cultivation effect in each stage of group 1 is superior to group 2, adopting the seed of Dendrobium chrysotoxum as group training material, the required cycle is far smaller than the cycle time adopting Dendrobium chrysotoxum tender shoots to need as group training material, and the cultivation cycle of group 1 is relative to group 2, shorten the cultivation time of 2 years, while being effectively ensured the Dendrobium chrysotoxum seedling that high-quality is provided, also meet market Production requirement；
Group 1 compares with group 3, group 4, group 5, group 6, group 7, the effect that the culture medium of group 1 produces is best, group 3, group 4, group 5, group 6 have lacked certain some raw material components respectively, culture medium lacks a certain raw material components, then can affect the cultivation effect in some stage, the cultivation poor effect reached, it may thus be appreciated that, group 1 is, according to what certain principle screened, Herba Dendrobii has fine induction, can in the good characteristic situation keeping former plant, in not variation situation, appreciation rate is high, grows fast excellent culture medium.
Although embodiment of the present invention are disclosed as above, but listed utilization that it is not restricted in description and embodiment, it can be applied to various applicable the field of the invention completely, for those skilled in the art, it is easily achieved other amendment, therefore, under the general concept limited without departing substantially from claim and equivalency range, the present invention is not limited to specific details.
Claims (4)
1. a Dendrobium fimbriatum Hook. seed tissue cultivates method for quickly breeding, it is characterized in that, taking Dendrobium fimbriatum Hook. seed and carry out in-vitro inducing cultivation as material, obtain the tissue culturing seedling of Dendrobium fimbriatum Hook., described Dendrobium fimbriatum Hook. seed tissue is cultivated method for quickly breeding and is comprised the following steps:
Step one, by the Dendrobium fimbriatum Hook. planting seed of pretreatment on inducing culture, cultivate 7-15 days, obtain Dendrobium fimbriatum Hook. budlet, wherein, described inducing culture is: with MS-H for minimal medium, add 0.5-3.0mg/L heteroauxing, 1.0-5.0mg/L naphthalene acetic acid, 0.1-1.0mg/L Thidiazuron, 25-40g/L sucrose and 3.0-5.0g/L agar, and the pH value adjusting inducing culture is 5.6-6.0；
Step 2, Dendrobium fimbriatum Hook. budlet is placed in subculture medium, cultivate 30-40 days, obtain Dendrobium fimbriatum Hook. group training differentiation Seedling, wherein, described subculture medium is: with MS for minimal medium, add 0.2-2.0mg/L kinetins, 2.0-4.0mg/L6-benayl aminopurine, 1.0-4.0mg/L naphthalene acetic acid, 0.5-3.0mg/L indolepopionic acid, 0.1-2.0mg/L Thidiazuron, 25-40g/L sucrose and 3.0-5.0g/L agar, and the pH value adjusting subculture medium is 5.6-6.0；
Step 3, by the training differentiation transplantation of seedlings of Dendrobium fimbriatum Hook. group in root media, after cultivating 30-40 days, being transferred under nature light and irradiate cultivation 40-50 days, it is thus achieved that Dendrobium fimbriatum Hook. is taken root tissue cultured seedling；Wherein, described root media is: with 1/2MS for minimal medium, adding 5-20% Fructus Musae, 0.1-4.0mg/L naphthalene acetic acid, 2.0-4.0mg/L6-benayl aminopurine, 2-5% sucrose and 0.2-0.5% activated carbon, and the pH value adjusting root media is 5.8-6.2；
Step 4, tissue cultured seedling of being taken root by Dendrobium fimbriatum Hook. are transplanted to seedbed, and described medium of seedling bed is: 20-30 bark, 10-20 weedtree, 20-30 Vermiculitum and 10-15 perlite, treat that the growth of Dendrobium fimbriatum Hook. seedling is stable, transplant to booth.
2. Dendrobium fimbriatum Hook. seed tissue cultivates method for quickly breeding as claimed in claim 1, it is characterized in that, the pretreatment of Dendrobium fimbriatum Hook. seed described in step one is, first by Dendrobium fimbriatum Hook. outstanding achievement with the 80-85% alcohol solution dipping 20-40 second, sterilize 10-20 minute with mercuric chloride again, cut the outstanding achievement of Dendrobium fimbriatum Hook. after cleaning up, obtain the Dendrobium fimbriatum Hook. seed of pretreatment.
3. Dendrobium fimbriatum Hook. seed tissue cultivates method for quickly breeding as claimed in claim 1, it is characterized in that, tissue cultured seedling of being taken root by Dendrobium fimbriatum Hook. in step 4 is transplanted to before seedbed, and the culture medium of tissue cultured seedling root of first being taken root by Dendrobium fimbriatum Hook. cleans up, dry steam, then transplant to seedbed.
4. Dendrobium fimbriatum Hook. seed tissue cultivates method for quickly breeding as claimed in claim 1, it is characterized in that, described step one is to step 3, and Dendrobium fimbriatum Hook. condition of culture in culture medium is: cultivation temperature is 28-32 DEG C, intensity of illumination is 2500-3000lux, and light application time is 13-15h/d.
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