CN101803515A - Method for rapidly growing and cultivating dendrobium officinale - Google Patents
Method for rapidly growing and cultivating dendrobium officinale Download PDFInfo
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Abstract
The invention discloses a method for rapidly growing and cultivating dendrobium officinale, which comprises the following steps of: taking back mature shadow fruit seeds in the country of origin for wild dendrobium officinale, and placing the seeds in a clean culture dish; after natural crack, pouring the seeds into a clean beaker, sealing the clean beaker, and storing the beaker in a low temperature box; performing a certain sterilization procedure on the stored seeds, placing the seeds in an MS minimal medium, performing seed germination cultivation, wherein the seeds gradually become green and form protocorm, and observing that needles appear in the protocorm by using magnifying glass; transferring the protocorm into the MS minimal medium, performing protocorm proliferation and differentiation culture, observing that plantlets are differentiated from the protocorm in a bottle along the production of partial new protocorm; and transferring the plantlets into the MS minimal medium, and performing strong seedling and rooting culture, seedling hardening and transplanting. The survival rate is over 97 percent without the seedling hardening process, and the yield is doubled. The growth speed is improved by 50 percent, the content of each component is equivalent to that of the wild dendrobium officinale, and the contents of amino acid and microelement are higher than those of the wild dendrobium officinale.
Description
Technical field
The invention belongs to a kind of dendrobium candidum breeding field, more specifically relate to a kind of method of rapidly growing and cultivating of dendrobium candidum.
Background technology
The celestial grass in the dendrobium candidum popular name human world; main component is alkaloid and polysaccharide; also be rich in amino acid; also contain abundant trace element. modern clinical practice and medicinal studies confirm that. dendrobium candidum is to pharyngolaryngitis; gastrointestinal disease; cataract; angiocardiopathy; diabetes and inhibition tumor growth have significant curative effect; crossing inside and outside market demand constantly increases; price soars all the way. because the growth of wild dendrobium candidum is slow; yield poorly; adding people constantly excavates; cause wild dendrobium candidum to face withered exhausted state. can't meet the need of market. the endangered species of treasuring of one class focused protection classified dendrobium candidum as by middle traditional Chinese medicines circle; and encouraging artificial commerial growing dendrobium candidum. some scientific research institutions of China and business unit explore and test the group training and the artificial commerial growing technology of dendrobium candidum at present; obtained certain achievement; but group training and culture time have used chemistry; chemical industry; raw materials such as agricultural chemicals; remain at the seedlings of Dendrobium officinale reproductive speed slow; tissue cultivating seedling; it is low to practice the transplantation of seedlings survival rate; growth rate is slow; output is not high; the residue chemistry composition is arranged; subject matters such as active ingredient is low have seriously restricted the dendrobium candidum industrialization; large-scale development.
Summary of the invention
The object of the present invention is to provide a kind of method for rapidly growing and cultivating of dendrobium candidum; from the tissue cultivating seedling to the cultivation; be not completely cured when guaranteeing seedling replanting, survival rate height, fast, high, the no residue chemistry composition of output of growth. the cultivation bed is ventilative, drainage has satisfied the characteristic that the dendrobium candidum natural substrates is grown nonparasitically upon another plant, the dendrobium candidum method for rapidly growing and cultivating of realization economized, scale, high yield.
To achieve these goals, the present invention adopts following technical measures:
The present invention through certain disinfectant program, carries out seed germination with the dendrobium candidum seed in homemade medium, protocorm increment differentiation, strong sprout, culture of rootage, and the time height of seedling degree length that needs 90 days approximately is to more than the 5cm.
Traditional seedlings of Dendrobium officinale tissue culture technology, be to adopt MS minimal medium+different hormones (a-methyl, indolebutyric acid, benzyladenine etc.) proportioning+organic additives (murphy juice, bananas juice etc.)+agar, but the hormone that it is selected, composition and ratio with organic additive, result of use is not ideal, train from the dendrobium candidum seed group and can transplant seedlings (more than the 5cm) and need 8-11 month approximately. because the medium that uses all is to lump, the conglomeration raw material tears low, the poor growth of root system, survival rate that should damage seedling when planting transplantation of seedlings.
The quick multiplication technique of dendrobium candidum provided by the invention, what need not the employing of hormone and organic additive is 140 order fine sands, only needs 90 days from the dendrobium candidum seed to cultivating into seedlings (more than the 5cm), has shortened the seedling group training time greatly, has reduced cost.
A kind of method for rapidly growing and cultivating of dendrobium candidum, the steps include: that it comprises that the oblique tissue cultivating seedling of the quick multiplication technique of the oblique seedling of iron sheet stone, iron sheet stone practices seedling technology, the oblique transplantation technique of iron sheet stone and the oblique collecting method of iron sheet stone, it is characterized in that the quick multiplication technique of the oblique seedling of iron sheet stone wherein is made up of following steps:
With the shady fruit seed of the oblique original producton location of wild iron sheet stone maturation back and forth after, be placed in the thousand clean culture dishes, treat its natural cracking after, seed poured in the clean beaker seal, deposit in the cryogenic box; With the certain disinfectant program of seed process that stores, put in the medium of MS minimal medium ten a-naa 0.5-0.9mg/L+ murphy juice 22-28W/V+ agar 8-11g/L, carrying out seed germination cultivates, cultivation temperature 24-27C, illumination every day 8 hours, light intensity 1200-1600lx, seed changes green gradually after 16 days, formed protocorm in ten months, and saw, can see that protocorm has needle to occur with magnifying glass; Protocorm is changed in MS minimal medium+6-card base gland howl mountain range 0.5-0.9mg/L+ murphy juice 22-28W/V+ agar 8-11g/L medium, carry out protocorm increment, differentiation culture, can be observed a bottle interior protocorm in 2 months and differentiate seedling, about high 1cm, simultaneous has the new protocorm of part to produce; Any change over to growing sturdily for a short time in the medium of MS minimal medium ten expansion naa 0.4-0.6mg/L+ murphy juices 20 V/V+ agar 8g/L, carrying out strengthening seedling and rooting cultivates, can be observed the middle seedling that seedling develops into 2.0-3.5cm in 2 months, Shen Miaozai is transferred in the same medium, carrying out seedlings again cultivates, can turn out the above seedlings of 5Cm height in 2 months, but the seedlings bottle outlet is practiced seedling, transplanting; The oblique tissue cultivating seedling of iron sheet stone is wherein practiced the seedling technology; Practicing seedling matrix is wood shavings+perlite+soddy soil, and volume ratio is H1: 1; Practicing seedling-growing method is that white silk seedling container is the high small flower of 7cm, at the fine stone of the shop, bottom of flowerpot diameter 1-2cm, thickness 3cm; The wood shavings, perlite, the soddy soil mixed-matrix that with volume ratio are 1: 1: 1 again are put on the fine stone, thickness 3cm, every basin is put 10 young plants, push mixed-matrix aside an alveole, put down gently the coring of plant in nest and cover, outside the base portion of plant is exposed to, water normal root water one time, control temperature, illuminance, moisture, humidity well; Slender of white silk is that common green house all can be used as Lian Miaopeng, and temperature 20-25 ℃ of air humidity practicing seedling is 75-85%, and water supply in media is profit, and matrix is crossed a wet or inclined to one side hundredweight can have a strong impact on experienced shoot survival percent; Illumination is the scattered light of normal illumination 1/3, light intensity 1800-2500lx, and practicing seedling season be annual spring, Qiu Liangji, practices seedling time 4-5 month; The oblique transplantation technique of iron sheet stone wherein is: transplanting medium is the big rubble of diameter 3-5cm+diameter 1-3cm fine stone+tongue Xue, and volume ratio is 4: 2: 1; Transplanting condition is that the common plastics booth all can be used as the transplanting booth, should dig around the booth and establish the drain ditch, and the canopy planted agent lays cultivation bed, the growth temperature 22-28 of transplanted seedling ℃; Air humidity 75-85%, water supply in media is profit; Illumination is the scattered light of normal illumination 1/3, and light intensity 2000--3200lx, transplanted seedling are annual spring, Qiu Liangji season;
Transplanting method is:
1, lays the big rubble of diameter 3-5Cm at cultivation bed bottom, thickness 8cm;
2, on big rubble, lay the fine stone of diameter 1-3cm, thickness 4cm;
3, the tongue evil after sterilization is dried, water soaked 30 minutes earlier, extracted that to put into frame of plastic stand-by again;
4, according to the ranks distance of 12 * 15Cm, push fine stone aside the worker alveole, every nest is put the 4-6 young plant;
5, the coring of plant is let go the fine stone face that gently is affixed in the nest, sporadicly spread the tongue evil that any is got ready around coring, the mansion fine stone covers coring gently again, outside the base portion of plant is exposed to;
6, plant seedling, watered normal root water one time; The oblique collecting method of iron sheet stone wherein is:
The oblique tissue cultivating seedling of iron sheet stone is after practicing seedling, transplanting, and 1.5-2 can gather, and 5-6 continuously can gather; Be annual December-January picking time, gathers should stop in preceding 1 month to plant fertilising and medication, should take when gathering old stay new, adopt and stay little principle greatly.
The fruit seed germination medium culture that the applicant obtains by the oblique maternal plant artificial pollination of iron sheet stone with robust growth, seed germination becomes protocorm and further forms seedling, with seedling test tube flowering medium culture, plant artificial pollination after will blooming, aseptic seeding behind the fruit maturation, make seed grow up to plantlet, plantlet is cultivated on the strong seedling culture base, the Cheng Miao that forms just can further cultivate into seedling through hardening, the survival rate of Cheng Miao can reach more than 95%, thereby has realized purpose of the present invention.Or be achieved through the following technical solutions the present invention: the oblique sapling multiplication method of iron sheet stone of the present invention, its feature comprises the steps:
A, choose robust growth when blooming iron sheet stone tiltedly (the disappointed go of Dendobi ware officinale Kimura et) maternal plant carry out artificial pollination, pollination back fruit maturation, with fruit as explant;
B, thoroughly use explant alcohol-pickled successively, the mercuric chloride solution sterilization, cut after the rinsed with sterile water, Powdered embryo is inoculated into the seed germination medium, and seed germination forms protocorm, and further forms seedling, described seed germination medium contains for every liter spends precious No. 1 1-1.5g albumen arm 0.5-2g coconut milk 50-100 disorderly, sucrose 15-30g agar 6-7g, active carbon 0.5-1g, all the other are the vitamin and the inositol composition of MS medium Shen;
C, with seedling test tube flowering medium culture, the plant of choosing robust growth after blooming carries out artificial pollination in vitro, cut behind the fruit maturation and carry out aseptic seeding, seed germination forms protocorm, and further form complete plantlet, described test tube flowering medium is every liter and contains 6-card base fat tinkling of pieces of jade 0.5-1.0mg, Cai's acetate 0.1-0.2 beer, and all the other are the MS medium;
D, plantlet is transplanted on the strong seedling culture base cultivates, be formed into and grow sturdily, described strong seedling culture base contains for every liter spends precious No. 1 1-1.5g to spend precious No. 2 white 0.1-0.3g of 1-1.5g albumen arteries and veins 0.5-2g hydrolysis junket firefly, bananas juice 50-150g, carrot juice 10-30g, apple juice lo-30g sucrose lo-20g agar 6-7g active carbon 1-2 naa 0,5-1.0mg, all the other are vitamin and inositol composition in the MS medium;
E, will become seedling, wash the medium of earning root then, in the matrix of mixing in 1: 1 by volume, cultivate, obtain seedling with sphagna or peat and Lan Shi at the natural daylight lower refining seedling;
Medium pH 5.2-5.4 in the above-mentioned steps (B)-(D), cultivation temperature (26 ± 2).
The described fructescence of step (A) is 110-130 days.
The described alcohol concentration of step (B) is volume fraction 70%-80% miaow preferably, soak time is 30-60 second preferably, the concentration of described mercuric chloride solution is mass fraction 0.1%-0.2% preferably, disinfecting time preferably 20-30 minute, the rinsing number of times of described sterile water preferably 4-5 time, the described treasured of spending can be buied for No. 1 from the market, in order to obtain more seedling in process of production, described protocorm can also be formed the line and staff control of bud and protocorms, on the shoot proliferation medium, can carry out shoot proliferation, the protocorms that forms can be proceeded propagation, the seedling of band root then enters the described cultivation of step (C), described shoot proliferation medium contains for every liter spends precious No. 1 1-2g, albumen knee 0.5-2g coconut milk 50-100mL, sucrose io-2og, agar 6-7g, 6-card base throat mountain range 0.5-1mL, active carbon 1-2g, all the other are vitamin and inositol composition in the MS medium.
The height of the described seedling of step (C) is preferably 2-3cm, and the described fructescence is 50-60 days.
The best high 3-4cm of the described one-tenth seedling of step (D) is whenever from having 2-3 bud, described spending precious No. 2.
The described hardening time of step (E) is preferably 8-12 days, and the illumination of described natural lighting is 1500-2000lx, the best 12-16 of light application time hour/day.
Step (B) and (D) precious No. 1 and spend and all be commercially available for precious No. 2 of spending of using of medium Shen.Again or adopt following technical measures:
1, dendrobium candidum fast seedling growing:
(1) condition of group training chamber:
With dendrobium candidum seed process routine disinfection program, put into 140 order fine sands, in the medium of humidity more than 96%, carrying out seed germination cultivates, cultivation temperature 24-26 ℃, illumination every day 8 hours, light intensity 1500-1800lx, about 2 all seeds change green gradually, form protocorm, can see that protocorm had needle to occur in 50 days in 45 days, can see that protocorm differentiated seedling in the dish in about 55 days, about high about 1cm, simultaneous has the new protocorm generation of part, can be observed the middle seedling that seedling develops into high 2-3cm in about 60 days.Left and right sides seedling can grow more than the 5cm in about 80 days.
(2) dendrobium candidum fast seedling growing:
With the certain disinfectant program of dendrobium candidum seed process, put into 140 order fine sands, should not lump, the seedlings branch tears the damage shoot root when planting, need not to practice seedling, carrying out seed germination in the medium of humidity 96% cultivates. and cultivation temperature 24-26 ℃, illumination every day 8 hours, light intensity 1500-1800ix about 2 all backs seeds change green gradually, formed protocorm in 45 days, can see that protocorm has needle to occur. can see that protocorm differentiated seedling in the dish in about 55 days, about high about 1cm, the protocorm that simultaneous is new produces, and about 60 can see that seedling develops into the seedling of high 2-3cm. and about about 80 days, seedling amount of money length is to more than the 5cm, but placing is transplanted to booth. seedlings transplanting method: in booth, adopt sawdust 8 orders 40%. pine bark 4mm * 4mm40%; Yellow sand 8 orders 20% for building, mix, be paved into wide 1500mm, the long cultivation bed of not limitting of thick 150mm. this method cultivation bed drainage, good permeability belong to original ecology cultivating matrix. and end product quality is pure. after the cultivation bed is put in order, seedlings are transplanted to a body, the ranks distance of spacing 120cm * 150cm. push matrix aside an alveole, every nest is put the 3-5 young plant, the coring of plant let go be affixed in the nest, around coring, sprinkle a small amount of cultivate material, be exposed to outer being advisable with the plant base portion, planted seedling after, water one time and follow water surely. control temperature, humidity, moisture, illuminance etc. well.
2, the dendrobium candidum transplanting of growing fast:
(1) transplanting condition:
General plastic tunnel all can be used as the transplanting booth; booth needs bedding to protect synovial membrane; the sunshade net. need drainage trenching all around; the canopy domestic demand is laid the cultivation bed. cultivation bedside 150mm; the operation aisle is stayed in the centre; 23-26 ℃ of transplanted seedling preference temperature; be higher than most 30 ℃; minimumly be not less than 8 ℃; air humidity 70-85% is advisable; water supply in media is good with profit. and water supply in media is crossed wet or overdrying all can influence plant strain growth and output. and illumination is normal illumination; the about 1800-3000lx of scattered light; to use the sunshade of sunshade net in summer and autumn. transplant the four seasons in season and all can cultivate. transplant and can gather in back 1 year, can gather throughout the year, get when gathering stay greatly little; the 5-6 that can gather continuously, but should stop to the plant spraying insecticide in previous month gathering.
(2) dendrobium candidum growing and cultivating method:
Candidum tissue culturing seedling is after cultivation in 90 days reaches more than the 5cm, plant has been in comparatively healthy and strong stable state, after the taking-up of group training basal disc. shake gently, group training base-material can come off automatically, need not tear, can not damage the root system of seedling, it is very fast to transplant the back growth. by making thorough investigation and study to wild dendrobium candidum ecosystem environment, through Multitest, its specified transplanting medium adopts sawdust 8 orders 40%, yellow sand 8 orders 20%, the ratio of pine bark 4mm * 4mm 40% (volume ratio) is mixed use, in conjunction with reasonably fertilising, water, controls such as illumination, can guarantee plant matrix permeable in 5-6, good air permeability the matrix substrate junction phenomena can not occur, and has certain characteristics such as preserve moisture. and survival rate of plant reaches more than 97%.
Though patent and the bibliographical information that produce relevant for the oblique seedling of iron sheet stone domestic morning, but do not see utilize test tube flowering to pollinate to breed iron sheet stone oblique for result's method.And method of the present invention, not only simple to operate, with low cost, and can shorten maturing stage of the oblique fruit of iron sheet stone widely, efficient and do not have a damage by disease and insect, can the anniversary produce the oblique seed of iron sheet stone, the transplanting survival rate of Cheng Miao can reach more than 95%, thereby opens up a new way for the breeding of the oblique seedling of iron sheet stone.The four seasons finished product of can gathering. the 5-6 that can gather continuously, year per mu yield is more than 500 kilograms.
Embodiment
Following examples are to further specify of the present invention, are not limitations of the present invention.What used the embodiment Shen spends precious No. 1 (HYPONeX 1) and spends precious 2-(HYPONeX 2) to be Taiwan produced in USA platform and gardening enterprise stock action Co., Ltd packing product.
Embodiment 1:
The maternal plant of choosing robust growth when blooming carries out artificial pollination, and the back 120 days fruit maturations of pollinating are used for sowing with fruit as explant.During sowing, with explant use successively volume fraction 7 alcohol-pickled 30 seconds, sterilization is 20 minutes in the mercuric chloride solution of mass fraction 0.1%, rinsed with sterile water 4 times, cut fruit then, (every liter contains and spends precious No. 1 1.5g albumen knee 0.5g coconut milk 50mL in the medium with transfer needle the Powdered embryo of yellow-white to be inoculated into the seed germination medium, sucrose 15g, agar 6g " active carbon weir; all the other are the vitamin and the inositol composition of MS medium Shen) on; seed germination after 10 days, form protocorm, form seedling after 80 days.
(every liter contains 6-card Ji Sangling (6-BA) 0.5mg to the seedling 100 strain rolling bottles that 2-3cm is high to the test tube flowering medium, Cai's acetate (NAA) 0.1mg, all the other are the MS medium) on, 20 strains are bloomed in the time of 45 days, flowering rate can reach 20 and copy, choosing in vitro, the plant of robust growth carries out artificial pollination in vitro, 55 days fruit maturations, come fruit down, fruit need not sterilization, cuts fruit and carries out aseptic seeding, and left and right sides seed germination formed protocorm in 7 days, can form complete plantlet in the time of 80 days, the quantity of each fruit is more than 50,000 strains.
The intensive plantlet that aseptic seeding is obtained is transferred to the strong seedling culture base after separately, and (every liter contains and spends precious No. 1 1.5g to spend precious No. 2 1.5g peptone 0.5g, caseinhydrolysate 0.1g, bananas juice 50g, carrot juice 10g, apple juice 10g, sucrose 10g agar 6g, active carbon 1g, NAA0.5mg, all the other are the vitamin and the inositol composition of MS medium Shen) go up and cultivate, formed plant height 3-4cm, every clump of Cheng Zhuo that 2-3 bud arranged in 80 days.To become seedling to transfer to the natural daylight lower refining seedling 10 days, illuminance 1500-2000lx, illumination 12 hours/day, then it is taken out from the vial Shen, clean the medium of root, in the matrix of mixing in 1: 1 by volume, cultivate, keep suitably all over wind and enough humidity with sphagna and Lan Shi, 1000 smear and survive 950 strains, and the survival rate of transplanting all can reach 95%.Medium used in above-mentioned experiment is pH5.2-5.4,26 ℃ of cultivation temperature.
Embodiment 2;
The maternal plant of choosing robust growth when blooming carries out artificial pollination, and the back 110 days fruit maturations of pollinating are used for sowing with fruit as explant; During sowing, explant is used successively volume fraction 75% alcohol-pickled 40 seconds, the mercuric chloride solution Shen sterilization of mass fraction 0.15% 30 minutes, rinsed with sterile water 5 times is cut fruit then, (every liter contains and spends precious 1-snake in the medium with transfer needle the Powdered embryo of yellow-white to be inoculated into the seed germination medium, albumen refining 1g, coconut milk 80mL, sucrose 25g, agar 6.5g, active carbon 0.8g, all the other are vitamin and inositol composition in the MS medium) on, seed germination after 10 days, form protocorm, in order to obtain more seedling in the production process Shen, protocorm is formed the line and staff control of bud and protocorms, (every liter contains and spends precious hole 1g at the shoot proliferation medium, albumen refining 0.5g, coconut milk 50mL, sucrose 10g agar 6g, active carbon 1g, 6-taro base howl cry of certain animals 0.5mg, all the other are vitamin and inositol composition in the Ms medium) go up shoot proliferation, propagation is 4 times after 40 days, and the protocorms of formation is proceeded propagation, obtain the seedling with root, the quantity of each fruit is more than 50,000 strains.(every liter contains 6-BA 1.0mg NAA 0.2mg to the test tube flowering medium with seedling 100 rolling bottles of 2-3cm high-band root, all the other are the MS medium) on, 22 strains are bloomed in the time of 45 days, and flowering rate can reach 22 and copy, choosing in vitro, the plant of robust growth carries out artificial pollination in vitro, 50 days fruit maturations are adopted down fruit, and fruit need not sterilization, cut fruit and carry out aseptic seeding, left and right sides seed germination formed protocorm in 7 days, can form complete plantlet in the time of 80 days, and the quantity of each fruit is more than 50,000 strains.The intensive plantlet that aseptic seeding is obtained is transferred to the strong seedling culture base after separately, and (every liter contains and spends precious No. 1 1g, spend precious No. 2 1g, peptone 2g, caseinhydrolysate 0.3g, bananas juice 150g carrot juice 30g, apple juice 30g, sucrose 20g, agar 6.5g, active carbon 2g, all the other are vitamin and inositol composition in the MS medium for NAA 1.0mg) go up and cultivate, formed plant height 3-4cm in 70 days) every clump of Cheng Miao that 2-3 bud arranged.
To become seedling to transfer to the natural daylight lower refining seedling 8 days, illuminance 15oo-2ooolx, illumination 14 hours/day, then it is taken out from vial, clean the medium of root, in the matrix of mixing in 1: 1 by volume, cultivate, keep suitably ventilating and enough humidity with sphagna and Lan Shi, 1000 smear and survive 965 strains, and the survival rate of transplanting all can reach 96.5%.Used medium is pH 5.2-5.4 in above-mentioned experiment, 26 ℃ of cultivation temperature.
Embodiment 3:
The maternal plant of choosing robust growth when blooming carries out artificial pollination, and the back 130 days fruit maturations of pollinating are used for sowing with fruit as explant.During sowing, explant is used successively volume fraction 80 alcohol-pickled 60 seconds, sterilization is 30 minutes in the mercuric chloride solution of mass fraction 0.2%, rinsed with sterile water 4 times, cut fruit then, (the medium Shen contains for every liter and spends precious No. 1 1.5g, albumen refining 2g coconut milk 100mL with transfer needle the Powdered embryo of yellow-white to be inoculated into the seed germination medium, sucrose 30g, agar 7g, the active carbon fork, all the other are vitamin and inositol composition in the Ms medium) on, seed germination after 10 days, form protocorm,, protocorm is formed the line and staff control of bud and protocorms in order to obtain more seedling in the production process Shen, (every liter contains and spends precious No. 1 2g albumen hip 2g coconut milk 100mL at the shoot proliferation medium, sucrose 20g, agar 7g, 6-fabricbase howl mountain range 1.omg, active carbon 2g, all the other are vitamin and inositol composition in the medium of valley) go up shoot proliferation, propagation is 4 times after 40 days, and the protocorms of formation is proceeded propagation, obtain the seedling with root, the quantity of each fruit is more than 50,000 strains.The seedling 100 strain rolling bottles that 2-3cm is high are to test tube flowering medium (every liter contains 6-BA0.5mg, NAA 0.1mg, all the other are the MS medium), 40 strains are bloomed in the time of 45 days, it is 40 good that flowering rate can reach, and chooses in vitro that the plant of robust growth carries out artificial pollination in vitro, 60 days fruit maturations, adopt down fruit, fruit does not have the vegetarian sterilization, cuts fruit and carries out aseptic seeding, and left and right sides seed germination formed protocorm in 7 days, can form complete plantlet in the time of 80 days, the quantity of each fruit is more than 50,000 strains.The intensive plantlet that aseptic seeding is obtained is transferred to the strong seedling culture base after separately, and (every liter contains and spends precious No. 1 1.2g, spend precious No. 2 1.2g, albumen swelling 1g, caseinhydrolysate 0.2g bananas juice 100g carrot juice 20g apple juice 20g, sucrose 15g agar 7g active carbon 1.5g, all the other are the vitamin and the inositol composition of MS medium Shen for NAA0.8mg) go up and cultivate, 60 days formation plant height 3-4cm, every clump of Cheng Miao that 2-3 bud arranged.To become seedling to transfer to the natural daylight lower refining seedling 12 days, illuminance 1500-2000lx, illumination 16 hours/day, then it is taken out from vial, clean the medium of root, in the matrix of mixing in 1: 1 by volume, cultivate, keep suitably ventilating and enough humidity with peat and Lan Shi, 1000 smear and survive 980 strains, and the survival rate of transplanting all can reach 98%.
Used medium is pH 5.2-5.4 in above-mentioned experiment Shen, 26 ℃ of cultivation temperature.
Claims (1)
1. the method for rapidly growing and cultivating of a dendrobium candidum, the steps include: that it comprises that the oblique tissue cultivating seedling of the quick multiplication technique of the oblique seedling of iron sheet stone, iron sheet stone practices seedling technology, the oblique transplantation technique of iron sheet stone and the oblique collecting method of iron sheet stone, it is characterized in that the quick multiplication technique of the oblique seedling of iron sheet stone wherein is made up of following steps:
With the shady fruit seed of the oblique original producton location of wild iron sheet stone maturation back and forth after, be placed in the clean culture dish, treat its natural cracking after, seed poured in the clean beaker seal, deposit in the cryogenic box;
With the certain disinfectant program of seed process that stores, put in the medium of MS minimal medium ten a-naa 0.5-0.9mg/L+ murphy juice 22-28W/V+ agar 8-11g/L, carrying out seed germination cultivates, cultivation temperature 24-27C, illumination every day 8 hours, light intensity 1200-1600lx, seed changes green gradually after 16 days, formed protocorm in ten months, and saw, can see that protocorm has needle to occur with magnifying glass; Protocorm is changed in MS minimal medium+6-card base gland howl mountain range 0.5-0.9mg/L+ murphy juice 22-28W/V+ agar 8-11g/L medium, carry out protocorm increment, differentiation culture, can be observed a bottle interior protocorm in 2 months and differentiate seedling, about high 1cm, simultaneous has the new protocorm of part to produce;
Any change over to growing sturdily for a short time in the medium of MS minimal medium ten expansion naa 0.4-0.6mg/L+ murphy juices 20 V/V+ agar 8g/L, carrying out strengthening seedling and rooting cultivates, can be observed the middle seedling that seedling develops into 2.0-3.5cm in 2 months, Shen Miaozai is transferred in the same medium, carrying out seedlings again cultivates, can turn out the above seedlings of 5Cm height in 2 months, but the seedlings bottle outlet is practiced seedling, transplanting;
The oblique tissue cultivating seedling of iron sheet stone is wherein practiced the seedling technology; Practicing seedling matrix is wood shavings+perlite+soddy soil, and volume ratio is Hl: 1;
Practicing seedling-growing method is that white silk seedling container is the high small flower of 7cm, at the fine stone of the shop, bottom of flowerpot diameter 1-2cm, thickness 3cm; The wood shavings, perlite, the soddy soil mixed-matrix that with volume ratio are 1: 1: 1 again are put on the fine stone, thickness 3cm, every basin is put 10 young plants, push mixed-matrix aside an alveole, put down gently the coring of plant in nest and cover, outside the base portion of plant is exposed to, water normal root water one time, control temperature, illuminance, moisture, humidity well;
Slender of white silk is that common green house all can be used as Lian Miaopeng, and temperature 20-25 ℃ of air humidity practicing seedling is 75-85%, and water supply in media is profit, and matrix is crossed a wet or inclined to one side hundredweight can have a strong impact on experienced shoot survival percent; Illumination is the scattered light of normal illumination 1/3, light intensity 1800-2500lx, and practicing seedling season be annual spring, Qiu Liangji, practices seedling time 4-5 month;
The oblique transplantation technique of iron sheet stone wherein is: transplanting medium is the big rubble of diameter 3-5cm+diameter 1-3cm fine stone+tongue Xue, and volume ratio is 4: 2: 1;
Transplanting condition is that the common plastics booth all can be used as the transplanting booth, should dig around the booth and establish the drain ditch, and the canopy planted agent lays cultivation bed, the growth temperature 22-28 of transplanted seedling ℃; Air humidity 75-85%, water supply in media is profit; Illumination is the scattered light of normal illumination 1/3, and light intensity 2000--3200lx, transplanted seedling are annual spring, Qiu Liangji season;
Transplanting method is:
1), at the big rubble of cultivating bed bottom laying diameter 3-5Cm, thickness 8cm;
2), on big rubble, lay the fine stone of diameter 1-3cm, thickness 4cm;
3), the tongue evil after sterilization dried, water soaked 30 minutes earlier, extracted that to put into frame of plastic stand-by again;
4), according to the ranks distance of 12 * 15Cm, push fine stone aside the worker alveole, every nest is put the 4-6 young plant;
5), let the coring of plant go gently be affixed in the nest fine stone face, sporadicly spread the tongue evil that any is got ready around coring, the mansion fine stone covers coring gently again, outside the base portion of plant is exposed to;
6), planted seedling, water normal root water one time; The oblique collecting method of iron sheet stone wherein is:
The oblique tissue cultivating seedling of iron sheet stone is after practicing seedling, transplanting, and 1.5-2 can gather, and 5-6 continuously can gather; Be annual December-January picking time, gathers should stop in preceding 1 month to plant fertilising and medication, should take when gathering old stay new, adopt and stay little principle greatly.
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