930 results about "Murashige and Skoog medium" patented technology

The invention discloses a hormone-free tissue culture method for dendrobium candidum, which comprises four tissue culture stages including germinating of seeds of the dendrobium candidum into spherical stems, multiplication and differentiation, and rootage and sound seedlings at stem sections; culture media used at the four stages include a hormone-free MS (Murashige and Skoog) culture medium, a hormone-free MS basal culture medium plus 15-20 g / l mashed potato culture medium, a hormone-free MS basal culture medium plus 15-20 g / l mashed banana plus 15-20 g / l mashed potato culture medium and a hormone-free 1 / 2 MS basal culture medium plus 15-20 g / l mashed banana plus 15-20 g / l mashed potato culture medium. The polysaccharide content and total alkaloid content in the culture of the dendrobium candidum obtained by hormone-free culture are increased compared with hormone-containing culture; the growth state of the dendrobium candidum is normal; the genetic diversity of the plant variety is protected; with the adoption of the method, the tissue culture of the dendrobium candidum is more convenient, green and environmentally friendly, and the resources are saved.

The invention discloses an isolated rooting culture method for a fir clone. The method comprises the following steps of: selecting basal sprouts of a trunk of an original plant of the excellent fir clone as a tissue culture material, inoculating the basal sprouts to a successive multiplication culture medium to perform successive multiplication culture, inducing and differentiating adventitious buds, and transferring the adventitious buds to an MS (Murashige and Skoog) culture medium to perform elongation culture; when the adventitious buds are grown to 2 to 3 centimeters, transferring the adventitious buds to an induced rooting culture medium, and performing culture to obtain rooted test tube plantlets; and transplanting the rooted test tube plantlets, and mixing loess and peat soil in a ratio of 3:1 to form s transplanting matrix. Compared with a fir tissue culture method in the prior art, the isolated rooting culture method for the fir clone has the advantages that: the rooting difficulty problem of tissue culture plantlets of different excellent clones of fir under the tissue culture condition is effectively solved, a stable and efficient tissue culture system of the fir is established, the highest rooting rate can reach 100 percent (onion 020), and a technical support is provided for quickening the industrialized development of the fir tissue culture plantlets.

The invention relates to a method for quickly propagating high-quality liquorice seedlings. The method is realized by sequentially comprising the following steps of: firstly, washing seeds with running water, sterilizing the seeds with 75% alcohol and washing 3-4 times with sterile water; then sterilizing the seeds with 0.1% corrosive sublimate, and washing 3-4 times with sterile water; then inoculating the seeds to an MS (Murashige and Skoog) culture medium for culturing aseptic seedlings for 1 month; then cutting stalks of the aseptic seedlings; chopping the stalks into stalk sections with the length of 0.5cm; and inoculating into a 0.5mg / L MS+KH2PO310mg / L+NAA culture medium to be cultured continuously. The method for quickly propagating the high quality liquorice seedlings has the advantages that the rooting rate of the aseptic seedlings reaches 100%, aseptic liquorice nursery stocks can be quickly propagated, the cycle is short, and the method is not influenced by factors, such as seasons, weather, natural disaster and the like and can realize industrial production. By adopting the technology, many problems of low propagation capability, shortage of resources, pesticide residue and the like of a medicinal plant can be solved. Since the technology for quickly propagating aseptic liquorice seedling stalks is firstly utilized in Gansu Province, in China, novel liquorice germ plasms in Gansu can be quickly propagated and meet market requirements and have strategic meanings in the aspects of preventing wind, stabilizing sands, protecting ecological environment and germ plasm.

The invention relates to a tender stem callus plant regeneration induction culture medium for lonicera caerulea and solves the problems of low plant differentiation rate and high culture medium cost problem of the auxiliary bud or stem tip culture, which is the main tissue culture of lonicera caerulea. The culture medium consists of a large amount of macroelements, microelements and hormone, wherein the hormone consists of 6-benzyl aminopurine at a concentration of 1.0 to 2.0mg / L and indolebutyric acid or naphthylacetic acid at a concentration of 0.05 to 0.2mg. When the LD1 culture medium of the invention is used, the tender stems of lonicera caerulea can be easily induced to form calla, and the induction rates of the calla are all over 90 percent; and the probability of the induction of the calla into plants is high, the plant differentiation rate is up to 50 to 82 percent which is 66.6 to 173.3 percent higher than that of the conventional Murashige-Skoog (MS) culture medium, and the cost is saved by 45.1 percent compared with the reference MS culture medium.

The invention provides a rapid propagation technology of Paris polyphylla Smith. The method comprises the following steps of: culturing tissues of the Paris polyphylla Smith such as rhizome sections and the like on a murashige and skoog (MS) culture medium, exactly taking a rhizome section with 1 to 2 buds as an explant, disinfecting, inoculating the explant into an MS plus 0.5mg / L 6-benzylamino adenine (BA) plus 0.4mg / L naphthylacetic acid (NAA) culture medium for culturing, ensuring the germination of the buds within about 6 to 8 days after the inoculation and the formation of seedlings with leaves within about 10 days, and cutting the seedlings and inoculating the seedlings into an MS plus 1.5mg / L 6-BA plus 0.4mg / L indoleacetic acid (IAA) culture medium to ensure that the high-speed increase is maintained and the multiplication factor is more than 5. MS plus 1.5mg / L 6-BA plus0.4mg / L IAA plus 0.5mg / L NAA is selected as a rooting culture medium, the rooting rate is over 90 percent, and an intact plant can be formed within about 25 days. A test tube seedling is transplanted into sand, the water retention and moisture retention are noticed, the transplant survival rate is over 90 percent after the seedling is transplanted for 1 month, and a set of high-frequency stable regeneration system is established. The method has the advantages of high stability, simple operation, high propagation speed, low production cost, industrialization level and the like.

The invention provides a method of rapid breeding of kiwi fruit with large seed by tissue culture. The invention comprises steps of selecting and treating explant, inducing culture medium, root culturing and replanting, etc. The invention takes MS culture medium as basis and adds BA, NAA, GA, ZT and other external plant growth regulator to induce the growth of axillary bud of the said kiwi fruit and strengthen sprout and root. The root growth rate is as high as 100%, the average bud height is 6.28 cm and the average replanting survival rate is 91% after 35 days culture. The invention solves the technique problem of rapid breeding of said kiwi fruit and provides a method of short period, high breeding rate and low cost for the industrial production of kiwi fruit with large seed.

The invention discloses a quick propagation method for lycoris chinensis. The method comprises the following steps of: taking embryos of the lycoris chinensis, and inoculating the embryos to an induction medium to induce callus; transferring the callus to an MS (Murashige and Skoog) culture medium containing sucrose, naphthyl acetic acid (NAA) and 6-benzyl aminopurine (6-BA) to induce adventitious buds; transferring the adventitious buds to the MS culture medium containing the sucrose, the NAA and the 6-BA to induce bulblets; transferring the expanded bulblets to an MS culture medium containing agar and NAA, and performing induced rooting under illumination; and transferring the rooted bulbs to a transplanting medium of perlite and humus. According to the method disclosed by the invention, the embryos of the lycoris chinensis are used as explants, the explants are induced to generate the callus, then the adventitious buds and the bulblets are induced, and plant regeneration is finally completed, so that a new path is increased for quick propagation of tissue culture of the lycoris chinensis. The method has high propagation coefficient, millions of bulblets can be produced in one year, and an effective path can be provided for quickly propagating seed bulbs; and the test tube plantlets have high differentiation synchronizing degree and good consistency, and the method has high large-scale production development value and good economic prospect.

The invention discloses a method for optimizing a potato isolated culture one-step seedling culture medium. The culture medium makes leaves of a potato test tube plantlet undergo isolated culture to form a seedling in one step. The one-step seedling culture medium takes an MS culture medium as a basal medium, and 6-benzyladenine, naphthyl acetic acid and 2,4-dichlorphenoxyacetic acid with different concentrations and combinations are supplied in 1L of the MS culture medium; adventitious buds and adventitious roots are directly differentiated through callus induction; and the potato regenerated seedling is obtained in one step. The callus obtained by induction in a primary culture medium is unnecessarily inoculated to a differential medium for regenerating the seedling. Compared with an isolated culture multi-step regenerated seedling method, the method of the invention has the advantages of simplified steps, short culture period, high repeatability, and high seedling survival rate.

The invention relates to a method for conserving chrysanthemum in vitro through reducing major element in the medium, which is special for the in vitro conservation of the chrysanthemum germplasm resource. The invention comprises the steps of: taking the suckers of the chrysanthemum as the explants, and axillary buds for subculture, and conserving on the 1 / 2MS, 1 / 2MS (1 / 2NH4+) or 1 / 4MS medium containing 6-BA0.3mg / L and NAA0.1mg / L. The survival rate for a 12-month conservation is 100%, 8 months longer than that of conservation on the normal MS medium. After the conservation material recovers to normal growth, the descendant is proved to have no genetic variance through the detection of POD, EST and ISSR. The invention uses the method of conserving chrysanthemum test-tube plantlet through reducing major element in the medium and identifies the genetic stability of the descendant, which proves that the plants grow normally after one-year conservation and the descendant has no genetic variation.

The invention discloses a modified medium for improving propagation of stems of Dendrobium officinale and a propagation method. The medium comprises MS (Murashige and Skoog) basal medium, cane sugar 20g / l, agar 6g / L, 6-BA (benayl aminopurine) 2mg / l, NAA (alpha-naphthaleneacetic acid) 0.2-0.5mg / l, banana mash 15-20g / l, potato mash 15-20g / l, and common tap water replacing distilled water. The modified MS medium is used to propagate the stems with axillary buds of Dendrobium officinale, alkaloid and polysaccharide content of Dendrobium officinale is evidently increased, and Dendrobium officinale grow normally. By the method, consumption of cane sugar is reduced, the tap water is used to replace the distilled water, and accordingly tissue culture technology for Dendrobium officinale is more convenient, more efficient and resource-saving.

The invention relates to a method for producing the lycoris by plant tissue culture, which comprises the steps of: (1) taking an MS (mass spectrometry) culture medium as a basic culture medium, and taking a 2, 4-D, IAA (indo acetic acid), NAA (naphthyl acetic acid), 6-BA (butyl acrylate) and IBA (iso butyl alcohol) as a regulated and controlled growth hormone preparation culture medium; (2) taking the bulb of the lycoris as an explant, and sterilizing; (3) vaccinating the explant of the bulb on a bulb inducing culture medium to culture in an inducing way so as to grow bulbils; (4) transferring the bulbils into a rooting culture medium to culture in a rooting way, so that the bulbils can independently absorb the nutrient elements from the soil and the matrix; (5) transferring the bulbils into the culture medium to culture; (6) opening an opening of a vaccination bottle to refine the seedling; withdrawing the bulbils, and transferring the bulbils into a nutrient cup which takes the vermiculite, the pearlite and the nutrient soil as the mixed matrix; and transferring the bulbils into a large tent, so that the survival rate reaches 94%-96%. The method is less in material taking, and economical in culturing; the culture condition can be manually controlled and is not influenced by the natural condition; the method is short in growing period, and high in propagation rate; and the quality of lycoris is guaranteed.

The invention relates to a method for rapidly propagating high-quality seedlings of Lycium ruthenicum Murr.. The method includes the steps as follows: firstly, seeds are washed with running water, sterilized with 75% (vol) alcohol and washed with aseptic water for 3 to 4 times; then, the seeds are sterilized with 0.1% (mass) mercuric chloride, washed with sterilized water for 3 to 4 times, and then inoculated in an MS culture medium to cultivate aseptic seedlings; and after the aseptic seedlings are cultivated for one month, the stems of the aseptic seedlings are cut off, and then cut into stem segments with the lengths of 0.5 cm and inoculated into a culture medium containing MSKH2PO413.5mg / LNAA0.1mg / L for cultivation. The method has the advantages that all the sterile seedling can take root, the aseptic seedlings of Lycium ruthenicum Murr. can be rapidly propagated, the period is short, the growth of the seedling is not influenced by factors of seasons, climate, natural disasters and the like, and the industrial production can be achieved; and by means of the technology, a plurality of problems of low propagation capacity, lack in resource, agriculture chemical residues and the like of a medicinal plant can be solved.

The application relates to a rapid propagation method of Bletilla striata. The method comprises the following steps: firstly culturing a pseudobulb section and other tissues of Bletilla striata on an MS (Murashige and Skoog) culture medium, cutting a rootstock section with 1-2 buds, which is served as an explant; disinfecting, and inoculating in a culture medium of MS+6-BA (0.5mg / L)+NAA (0.4mg / L) and culturing, wherein the buds germinate about 6-8 days after inoculation and grow into seedlings with leaves about 10 days after inoculation; cutting down the seedlings and transferring to a culture medium of MS+6-BA (1.5mg / L)+IAA (0.4mg / L) so as to maintain high-speed multiplication, wherein the multiplication coefficient is higher than 5; using MS+6-BA (1.5mg / L)+IAA (0.4mg / L)+NAA (0.5mg / L) as a rooting culture medium, wherein the rooting rate is higher than 90% and the seedlings can grow into complete plants only about 25 days; and transplanting test-tube plantlets into sandy soil and maintaining proper water content and moisture, wherein transplanting survival rate is higher than 90% one month after transplanting and a set of high-frequency stable regeneration system is established. The method disclosed by the invention has the advantages of being good in stability, simple and convenient in operation, high in propagation speed, low in production cost and reaching the industrialization level and the like.

The invention relates to a hormone-free tissue culture method for dendrobium officinale. The hormone-free tissue culture method comprises the following steps: a, inducing and culturing seeds; b, proliferating protocorms; c, rooting proliferated seedlings. In the step a and the step b, an adopted culture medium contains an MS (Murashige and Skoog) culture medium, 30g / L of white granulated sugar, 6g / L of agar and 100g / L of potatoes and the pH (Potential of Hydrogen) is 5.8-6.5; a culture medium adopted by the step c contains the MS culture medium, 30g / L of the white granulated sugar, 6g / L of the agar, 100g / L of the potatoes and 5g / L of active carbon, and the pH (Potential of Hydrogen) is 5.8-6.5. The method provided by the invention is simple and convenient in operation steps, and short in production period; the manpower and the material resources are saved; culture conditions are simple, the management is convenient and the space is saved; the types of the used culture mediums are few; the method is simple in preparation, reduces the production cost and is safe and environment-friendly; the cultured dendrobium officinale has high nutrition and medicinal component contents and better medicinal effects.

The invention discloses an asexual rapid propagation method for radix glycyrrhizae. The asexual rapid propagation method comprises the following steps: inoculating an explant of a radix glycyrrhizae stem section into a culture medium for cultivation to obtain an aseptic seedling; then cutting the aseptic seedling into a terminal bud explant, a stem section explant with one axillary bud and an explant with one axillary bud and a root, inoculating the explants into the same culture medium for cultivation to obtain a subculture-multiplicative aseptic seedling; and transplanting the subculture-multiplicative aseptic seedling into the culture medium to realize asexual rapid propagation for the radix glycyrrhizae. The formula of the used culture medium is that IBA with the concentration of 0.1mg / L is added to the MS culture medium, and the using amounts of KNO3 and NH4NO3 in the MS culture medium are halved respectively, and the pH value of the MS culture medium is 5.8. According to the asexual rapid propagation method, the root-inducing rate of the aseptic seedling is 100%, so that seedling emergence can be realized after 40 days, the seedling can be transplanted into a large field after 50 days, and the survival rate in the large field is higher than 80%. The asexual rapid propagation method can be suitable for rapid propagation of a plurality of excellent strain systems of a plurality of radix glycyrrhizae varieties, and has remarkable ecological, economic and social benefits in sustainable development of the radix glycyrrhizae.

The invention discloses a method for producing a Fujian jewel orchid by plant tissue culture, which comprises the following three procedures: selection and sterile treatment of plants, induced culture of cluster buds and large-scale culture. The method is technically characterized in that two different culture media are used in the induced culture procedure and the large-scale culture procedure, wherein the culture medium for the induced culture comprises an MS culture medium, sucrose of 10-50g / L, agar of 6-8g / L, activated carbon of 2.0-3.0g / L, naphthylacetic acid of 0.1-3.0mg / L and 6-benzylaminopurine of 0.1-5.0mg / L, and the pH value is 5.0-7.0; and the culture medium for the large-scale culture comprises a modified MS culture medium, calcium nitrate of 600mg / L, mashed banana of 30-50g / L, sucrose of 20-40g / L, agar of 6-8g / L, activated carbon of 2.0-3.0g / L, naphthylacetic acid of 1.0-2.0mg / L and 6-benzylaminopurine of 2.0-3.0mg / L, and the pH value is 5.0-7.0. Plant tissue culture can keep the good characters of the plants. The method has the advantages of easy operation, low production cost, high yield, good quality and no environment pollution, is suitable for industrial seedling raising and can realize mass production.

The present invention discloses the process of culturing adventitious root tissue of licorice, and belongs to the tissue culture of licorice as one kind of Chinese medicinal materials. The process of the present invention includes soaking licorice seed in solution with ethanol in 70-75 vol% and sodium hypochlorite in 2 vol% to sterilize and planting in MS culture medium to induce bacteria-free bud; inoculating the cut cotyledon and bud in MS culture medium containing different plant growth regulator to induce callus; transferring the callus growing for 20 days in MS culture medium with indolebutyric acid and kinin to induce adventitious root; and finally transferring the adventitious root to liquid culture medium containing the same plant growth regulator for fast growth. The present invention has simple process, high inducing rate, fast adventitious root proliferating speed and other features.

The invention discloses a manufacturing method of ecological moss blankets, comprising the following four steps: step 1. matrix cloth manufacturing: taking a non-woven fabric or a felt made of chemical fiber silks as a matrix cloth, wherein, the thickness of a single layer is 2-5mm, and the density is 500-1000g / m<2>; step 2. moss planting: collecting a moss from the field; washing the moss; directly inoculating on the matrix cloth or roughly smashing the moss, and scattering on the matrix cloth or spraying on the matrix by adding water; placing at a place with shade until growing and spreading into a sheet to cover on the matrix cloth so as to prepare into the moss blanket for later use; step 3. the moss blanket laying in three manners, namely horizontal laying, inclined laying and vertical laying; and step 4. watering and applying fertilizer: watering the moss blanket in horizontal laying and slight grade inclined layer by virtue of spraying; watering the moss blanket in vertical laying and inclined laying in a dropwise irrigating manner or a dropwise seeping manner by arranging a weeping pipe or a water seepage pipe on the matrix cloth, wherein the nutritive liquid needs not to be added on the moss blanket watered by lake water or pond water; and a leaf fertilizer is added on the moss blanket watered by running water in the room or a 0.01-0.02ppm MS (Murashige and Skoog) culture medium used for tissue culture is used as a nutritive liquid.

The invention discloses a method for making and storing artificial seeds of Dendrobium candicum Wall.ex Lindl., which comprises the following steps: (1) inducing protocorm; (2) performing liquid suspension culture and proliferation of the protocorm; (3) making the artificial seeds; and (4) storing the artificial seeds. Compared with the prior art, the invention has the advantages that the protocorm of the Dendrobium candicum Wall.ex Lindl. is proliferated through liquid suspension culture (1 / 2MS medium), the proliferation efficiency is obviously improved compared with that of the conventional solid medium (MS medium), and massive high-quality artificial embryos are provided for artificial seed production; sodium alga acid and a calcium nitrate solution are subjected to ion exchange and curing to form artificial seed coats, and the prepared artificial seed coats have regular shapes and low nutrient leakage rate compared with those prepared by reacting the sodium alga acid and the calcium nitrate solution in the prior art; and the artificial seeds have long storage life and high germination rate.

The invention provides a process for quickly propagating a large quantity complete oil tea clone plants with roots by tissue culture, which comprises the steps of secondary culture medium screening and hormone combination, seedling strengthening culture medium screening and hormone combination, root preprocessing, and rooting culture medium screening and hormone combination, wherein WPM+BA (Woody Plant Medium and Benzyladenine, 3-7mg / L), IBA (Indol Butyric Acid, 0.1-2mg / L) and ZT (Zeatin, 0.05-2mg / L) are used as secondary culture media for propagation, the propagation period is shortened to 30-40 days, and the propagation coefficient reaches 4.2-9.1; WPM+IBA (0.1-2mg / L) and biotin (0.5-3mg / L) are used as seedling strengthening culture media for strengthening seedlings, and seedlings can take root after 20-25 days; and after being treated with IBA, tissue culture seedlings which are 2-4cm in height are directly inoculated on the culture media of MS (Murashige and Skoog medium, 1 / 4-1) and AC (Activated Carbon, 200-600), the tissue culture seedlings take root and form complete plants after 25-3 0days, and the rooting rate reaches more than 85%.

The invention relates to a preparation method of an MS medium. The MS medium comprises macro-elements (a mother liquor 1), trace elements (a mother liquor 2), iron salt (a mother liquor 3) and organic components (a mother liquor 4). The preparation method is characterized in that Ca<2+> in the macro-elements is individually processed in the preparation process of the mother liquor 1 to prepare a storage liquor; in the preparation process of the mother liquor 2, sodium molybdate (NaMoO4.2H2O) is individually dissolved by distilled water at room temperature, then the dissolved sodium molybdate is poured into the trace element mother liquor 2; and in the preparation process of the mother liquor 3, FeSO4.7H2O and Na2-EDTA.2H2O are accurately weighed according to the consumption of 1000ml of the mother liquor, are respectively placed in a beaker with 500ml of distilled water, are heated and are continuously stirred to dissolve, heating is stopped until the solution approaches boiling, the obtained solutions are cooled, the cooled solutions are poured into reagent bottles, and the reagent bottles are put in a refrigerator for preservation. The preparation method of the MS medium solves the problems of difficult dissolving of sodium molybdate in the medium mother liquor preparation process and the precipitate generation or crystal precipitation in the mother liquor preparation and storage processes.

A regeneration and genetic transformation method for citrus includes immersing seeds in the solution of alcohol or mercuric chloride for disinfection, flushing with aseptic water, culturing in culture medium to obtain aseptic seedling, inducing the bud of check material, culturing in culture medium MS, adding growth regulator, transforming explant, using Agrobacterium to dipdys the intersegemental stem of said aseptic seedling, inducing resistant bud, adventitious bud grafting, and secondary grafting in greenhouse.

The invention relates to the field of flower propagation methods, in particular to the field of corm flower propagation methods. A thin layer culture method of tulips comprises the following steps of: longitudinally or transversely cutting tulip scales into thin layers of 0.1-5mm, and laying the thin layers on a culture medium; and inducing and proliferating the culture medium into an MS (Murashige and Skoog) culture medium, wherein 1.0-2.0mg of 6-BA (Benzyl Aminopurine), 0.1-1.0mg of NAA (Naphthyl Acetic Acid), 1.0-4.0mg of 2, 4-D (2,4-Dichlorophenoxyacetic acid), 20g of cane sugar and 6g of agaragar are added to the MS culture medium per L, the culture condition is that the temperature is 23+ / - 2 DEG C, the illumination intensity is 1,500-2,000Lx, and the illumination time is for 16h / d. The thin layer culture method of the tulips has the advantages and active effects: the tissue culture bulb inductivity reaches 100%, induced tissue culture seedlings grow vigorously, the source of the tissue culture seedlings is not limited by seasons, and the tissue culture scales are the better source of thin layer culture materials. Cutting modes cause different inductions on bulbs, and the differentiation rate and the bulb induction number of the scales which are transversely cut are higher than those of the scales which are longitudinally cut.

The invention relates to a method for regenerating plant fromarundo donax linn callus. The method comprises the following steps of (1) inducing callus: inoculating arundo donax linn tissue into culture medium containing1.0-5.0mg / l of 2,4-D (2,4-dichlorophenoxyacetic acid), carrying out dark cultivation for 3 days after inoculating, and then cultivating under the condition that the light period is 16 hours of illumination and 8 hours of darkness, and the illumination intensity is 2000Lx; (2) succesive transfer culture of callus: inoculating the callus into the culture medium containing 5.0mg / l of 2,4-D; (3) regenerating the callus plant: taking MS culture medium as a basic culture medium, and adding 0.5mg / L of IBA (indolebutyric acid) and 5.0mg / L of 6-BA (6-benzylaminopurine); (4) sprouting cultivation; (5) rooting cultivation: inoculating the plant into the MS culture medium in which 0-0.5mg / L of NAA (naphthylacetic acid) is added to root after sprouting; and (6) transplanting. A method for regenerating the plant from arundo donax linn callus is built by the invention; and expanding propagation of excellent arundo donax linn is facilitated.

The invention provides a fir tissue culture rooting method. Small fir multiplication subculture seedlings are cut and transplanted to a fir rooting culture medium for culturing and rooting tissue, and the fir rooting culture medium comprises an improved MS culture medium, 0.4-0.8mg / L of ABT1#, 0.1-0.5mg / L of IBA and 2-6mL / L of root sun diluent. After the fir rooting culture medium is adopted, the rooting time of firs is shortened to 16 to 22 days; after 22 days of culture, the highest rooting rate achieves 96 percent, and nursery stocks grow rapidly so as to decrease the culture cost of the seedlings and realize the industrialized production of the firs.

The invention mainly relates to a culture medium composition suitable for germ-free germination of orchid seeds and a method thereof. Various components of a basic culture medium and a seed germination culture medium are as follows: 500mg / L of calcium nitrate, 200mg / L of potassium chloride, 20-30g / L of sucrose or white sugar and 5-7g / L of agar, wherein the basic culture medium is selected from self-improved MS, i.e. on the basis of the original MS culture medium, 440mg / L of calcium chloride in the MS basic culture medium is replaced into 500mg / L of calcium nitrate and 200mg / L of potassium chloride; and the pH value is 5.0-5.6. The seed germination culture medium comprises the basic culture medium, 0.5-1.0mg / L of 6-BA, 0.5-1.0mg / L of NAA, 1.0g / l of active carbon and 50-100m / L of coconut milk. The invention has the advantages that: (1) the orchid group-cultured optimized culture medium has strong pertinence, good adaptability and high seed germination rate; (2) the method greatly shortens the breeding period of orchid so as to rapidly obtain new genetic improvement; and (3) the method is convenient for operation and reduces the pollution to the minimum extent.

The invention discloses a rapid proliferation method for anoectochilus formosanus tissue culture, which comprises the steps: cleaning and disinfecting an explant, inoculating the explant after cleaning and disinfecting in an MS culture medium, performing low light culture for one week, then performing strong light culture for one week, wherein the illumination time is 10 hours per day, and the temperature is 23+ / -2 DEG C; culturing for 40 days; inoculating the explants after culturing in a liquid culture medium of the MS culture medium, 1.4-1.6mg / L of 6-BA and 0.2-0.3mg / L of NAA in a ratio for enrichment culture under a dark culture condition, wherein the temperature is 23+ / -2 DEG C, and the culture period is 45 days; and by using a strong seedling culture medium as the MS culture medium, performing illumination culture, wherein the illumination time is 8 hours per day, the temperature is 23+ / -2 DEG C, the culture period is 12-15 days, and the ratio of a rooting culture medium is as follows: the content of MS is 1 / 2, the content of bananas is 60g / L, the content of active carbon is 1g / L, the content of casein hydrolysate is 0.5g / L, the illumination time is 10 hours per day, the temperature is 23+ / -2 DEG C, and the culture period is 3 months. According to the rapid proliferation method for the anoectochilus formosanus tissue culture, the industrialized rapid proliferation is realized, and the culture period is shortened.

The invention provides a method for efficiently breeding tissue culture seedlings of bletilla striata and a planting method of the bletilla striata. The method for efficiently breeding the tissue culture seedlings of the bletilla striata comprises the following steps: seeding bletilla striata seeds into a seed germination culture medium; carrying out cultivation and germination for 30-40d; and transferring the germinated seeds into a seedling culture medium to cultivate for 3-4 months, wherein the seed germination culture medium comprises 1 / 2 modified MS culture medium, 30-40g / L of potato juice and 20-30g / L of cane sugar; the seedling culture medium comprises the modified MS culture medium, 30-40g / L of a potato juice, 60-70g / L of a banana juice, 20-30g / L of cane sugar and 3.8-4.5g / L of agar powder. The planting method of the bletilla striata comprises the following steps: transplanting the tissue culture seedlings of the bletilla striata into a farmland; carrying out water and fertilizer management after transplanting; and carrying out weed control and diseases and pests prevention and control. The methods are liquid culture medium methods, and only need to transfer once. Compared with a traditional solid culture method, the methods are simple to operate, small in pollution, low in cost, high survival rate and short in culture cycle.

The present invention uses improved MS culture medium as basis, and adds the active carbon, indole-3-acetic acid, 3-indolebutyric acid and 6-benzylaminopurine as auxiliary agent to make tissue cultivation of two key stages of induced growth of bunch bud and seedling-strengthening and rooting of camllia chrysantha. The invention can be used for implementing industrial quick seedling cultivation.

The present invention provides one method of culturing Northern Cordyceps in a airlift bioreactor in large scale and separating and extracting Cordyceps extract. Seed for fermentation is cultured in MS culture medium and then transferred to 20 L bioreactor for large scale culture. The culture liquid in the bioreactor is then separated and purified to obtain product with Cordyceps extract content up to 46.25 % and purity of 99 %. The process of the present invention has low production cost and no environmental pollution.