CN105794438A - Gastrodia elata cultivation method - Google Patents
Gastrodia elata cultivation method Download PDFInfo
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- CN105794438A CN105794438A CN201410843167.4A CN201410843167A CN105794438A CN 105794438 A CN105794438 A CN 105794438A CN 201410843167 A CN201410843167 A CN 201410843167A CN 105794438 A CN105794438 A CN 105794438A
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Abstract
The invention relates to the technical field of traditional Chinese medicine material cultivation, and specifically relates to a gastrodia elata cultivation method. The method comprises the steps of (1) separating material selection and collection, (2) separation technology, (3) germination strain and armillaria mellea second-level strain (stock) cultivation, (4) germination strain and armillaria mellea third-level strain (cultivated strain) cultivation, and (5) sowing. With the method, gastrodia elata yield can be effectively improved, cost can be reduced, the harm of infectious microbes can be reduced, the quality of produced gastrodia elata can be improved, and growth period can be shortened. With the method, high-quality gastrodia elata products with high yield can be rapidly produced.
Description
Technical field
The present invention relates to Chinese crude drug planting technology field, specifically a kind of gastrodin cultivation method.
Background technology
Rhizoma Gastrodiae applicating history is long, at home and abroad has long enjoyed a good reputation.When the B.C. 100 Duo Nian Eastern Han Dynasty, first pharmacy monograph Shennong's Herbal of China then gives beginning load Rhizoma Gastrodiae, is classified as top grade, rhizoma gastrodiae by name.Rhizoma Gastrodiae is the famous and precious the most frequently used Chinese medicine of China, is also the famous authentic medicinal herbs in Guizhou, goes through version " Chinese Pharmacopoeia " and all gives and recording.It is the orchid family, Gastrodia herbaceos perennial, is distributed in whole nation most area.Rhizoma Gastrodiae is rich in gastrodine, gastrodia elata polysaccharide, vitamin A, glycoside, alkaloid, vanillyl alcohol, vanillin, succinic acid, cupreol, lymphatic temperament etc., and wherein gastrodine and gastrodia elata polysaccharide are main components.Its dry tuber, also known as Rhizoma Gastrodiae, is commonly use and famousr and precious Chinese medicine simply, and clinic is used for the diseases such as headache dizziness, numb limbs and tense tendons, infantile convulsion, epilepsy, tic, tetanus.Rhizoma Gastrodiae is born in the sylvan life that humus is more and moistening, chearance, border, and shrubbery and grass slope also have on the sunny side, height above sea level 400-3200 rice.With Bai Mo section fungus armillaria mellea and Mycen osmundicola symbiosis, just must can make seed germination, form ball stem, and be grown to serve as common Gastrodia Elata Stem Tuber.Mycen osmundicola provides nutrition for seed germination, and Armillaria mellea grows up to Gastrodia Elata Stem Tuber for protocorm bacterium provides nutrition.
Summary of the invention
The present invention provides a kind of production cost low, the gastrodin cultivation method that growth cycle is short.
Propose the specific scheme is that for solving above-mentioned purpose
1, selection and the collection of material are separated:
1. Armillaria mellea separates material: in rainy season in late fall, when Armillaria mellea sporophore is unearthed, and fresh, the form of gathering is complete, healthy and strong, cause harm without disease pest, will the word entity of maturation.Gastrodia Elata Stem Tuber: can gather, in 4-6 month, the Gastrodia Elata Stem Tuber that raw Armillaria mellea is more.2. Germination Strain separation material: the protocorm after collecting tuber of elevated gastrodia germination, the root in orchid, inducing culture Germination Strain sporophore.
2, isolation technics
Separation method: by Rhizomorph of Armillaria mellea or Gastrodia Elata Stem Tuber, Rhizoma Gastrodiae protocorm, the root of orchid washes surface earth respectively, then cuts the part tissue of needs, with aseptic water washing 3 times, puts into 0.1% mercuric chloride solution and soaks 1 minute；Use aseptic water washing 3 times again, wash away the medicinal liquid of remaining, be placed in sterile petri dish.The materials such as shoestring are cut into respectively segment or the fritter of 1 centimeter length, leaching a moment in penicillin liquid (20 mcg/ml), surface attachment water liquid is sucked with sterilizing filter paper, then it is inoculated on slant medium, put 25 DEG C of constant temperature culture, about 3～7 days, start to grow white hypha and shoestring respectively；During segregant entity, after aseptic water washing sterilization remaining medicinal liquid, exhaust the globule with sterilizing filter paper, then from cap, longitudinally cut entity open with dissecting knife, take its block organization of 0.5 centimetre at stem and cap intersection and be placed in PDA culture medium and cultivate.
Purifying agaric is cultivated: the mycelium that Germination Strain selection colony growth rate is more consistent, cuts mycelia with the inoculation of sterilization shovel together with culture medium, transfers to and cultivates in new culture medium, repeats until purifying agaric；Armillaria mellea when in culture medium, vaccination place has just produced shoestring branch, selects wherein to grow vigorous and young tender shoestring, intercepts long 2～3 millimeters with inoculation shovel and moves on slant mediums, cultivates under 25 DEG C of constant temperature, and shoestring is purification strain after covering with culture medium.
3, Germination Strain, Armillaria mellea second class inoculum (original seed) are cultivated
(1) Germination Strain second class inoculum is cultivated: medium component: the wood flour 70% of broad leaf tree, wheat bran or Testa oryzae 26%, sucrose 1%, Gypsum Fibrosum powder 1%, potassium dihydrogen phosphate 1.5%, the magnesium sulfate 0.5% such as green grass or young crops, birch, adds water appropriate.Above composition is fully mixed thoroughly, pinches culture medium with hands and can drip for spending, then bottle, beat the circular hole of diameter 1 centimetre at bottle center, as deep as culture medium 2/3 place.Build bottle cap, autoclaving (1.5 thousand grams/cm) 1 hour, inoculate after cooling.In transfer room, by the mycelia female kind together with about Semen Viciae fabae size, be connected in the circular hole of culture medium, one female plant can connect former in about 5 bottles, build the rearmounted 25 DEG C of culturing room of bottle cap and cultivate.
(2) Armillaria mellea second class inoculum cultivation: medium component is broad leaf tree wood sawdust 3 parts, 1 part of Testa Tritici and twig section, water is appropriate.During making, first the branch of thick fingers is cut into the segment of long 1.5 centimetres, water soaks fully water suction in 12 hours, then mix thoroughly with wood sawdust, Testa Tritici, add water drenched, load in 500 milliliters of wide mouthed bottles, often bottled enter branch section 50～60 sections, be advisable with the 4/5 of bottle capacity, add water a small amount of, autoclaving (1.5 thousand grams/cm) 1 hour after lid bottle cap, after cooling, access mother and plant, be placed in 25 DEG C of constant temperature culture 30 days～45 days.
4, Germination Strain, Armillaria mellea three-class strain (cultigen) are cultivated
(1) Germination Strain three-class strain is cultivated: taking Fagaceae fallen leaves 70%, wood flour 10%, Testa Tritici 15%, magnesium sulfate 0.5%, potassium dihydrogen phosphate 1.5%, carbamide 1%, sucrose 1%, Gypsum Fibrosum 1%, water is appropriate.Bottle after mixing thoroughly, autoclaving (1.5 thousand grams/cm) 1 hour, accesses second class inoculum, builds bottle cap, move into culturing room and cultivate after cooling.
(2) three grades of kinds of Armillaria mellea are cultivated: compost is identical with second class inoculum cultivation with cultural method.After autoclaving cools down, every bottle graft enters two grades of branch strain 1-2 sections, and 25 DEG C of constant temperature culture 30 days～45 days, Armillaria mellea silk and shoestring can cover with full bottle, can be used for cultivating bacterium branch and bacterium material.
5, sowing
(1) bacterium leaf seed dressing: 1. first by three grades of kinds of Germination Strain before sowing, by every square metre of consumption 4 bottles, takes out from seed bottle with the cleek of cleaning, puts in the seed dressing basin of cleaning, bacterium leaf is torn into individual；2. the tender fruit of Rhizoma Gastrodiae gathered and by every square metre, dehiscent fruit is sowed capsule 30～40, shakes out seed；3. seed is loaded planter box and is sprinkling upon on bacterium leaf, simultaneously with the dynamic bacterium leaf of handspring, seed is uniformly mixed on bacterium leaf, and is divided into two parts；
(2) bacterium sowing in bed kind: excavate bacterium bed during sowing, take out bacterium material, at the bottom of raking bed, the first wet fallen leaves of paving a thin layer, then the bacterium leaf divided is sprinkling upon on fallen leaves with 1 part, in statu quo set lower floor's bacterium material, 3～4 cm distance are stayed between bacterium material, blinding is put down to bacterium material, repave wet fallen leaves, spread another part of seed dressing bacterium leaf, put earthing 5-6 centimetre after bacterium material, bed top lid layer leaves moisturizing, then, seeding layer spreads one layer of cultured Armillaria mellea three-class strain, use 8-12 weight portion for every square metre, spread last layer clear water again and soak drenched broad-leaved leaves, every 40-60 weight parts water adds ground phosphate rock 0.3-0.6 weight portion, the upper fine sand covering 8-12cm or fertile soil, moisturizing.
The beneficial effects of the present invention is:
Can effectively improve the yield of Rhizoma Gastrodiae, reduce cost, the harm of the miscellaneous bacteria of minimizing, improve the quality of produced Rhizoma Gastrodiae, growth cycle can be shortened, reach quickly to produce the purpose of the sky ramie product of high yield, high-quality.
Detailed description of the invention
1, selection and the collection of material are separated:
1. Armillaria mellea separates material: in rainy season in late fall, when Armillaria mellea sporophore is unearthed, and fresh, the form of gathering is complete, healthy and strong, cause harm without disease pest, will the word entity of maturation.2. Germination Strain separation material: the protocorm after collecting tuber of elevated gastrodia germination, the root in orchid, inducing culture Germination Strain sporophore.
2, isolation technics
Separation method: Rhizomorph of Armillaria mellea washes surface earth, then cuts the part tissue of needs, with aseptic water washing 3 times, puts into 0.1% mercuric chloride solution and soaks 1 minute；Use aseptic water washing 3 times again, wash away the medicinal liquid of remaining, be placed in sterile petri dish.The materials such as shoestring are cut into respectively segment or the fritter of 1 centimeter length, leaching a moment in penicillin liquid (20 mcg/ml), surface attachment water liquid is sucked with sterilizing filter paper, then it is inoculated on slant medium, put 25 DEG C of constant temperature culture, about 3～7 days, start to grow white hypha and shoestring respectively；During segregant entity, after aseptic water washing sterilization remaining medicinal liquid, exhaust the globule with sterilizing filter paper, then from cap, longitudinally cut entity open with dissecting knife, take its block organization of 0.5 centimetre at stem and cap intersection and be placed in PDA culture medium and cultivate.
Purifying agaric is cultivated: the mycelium that Germination Strain selection colony growth rate is more consistent, cuts mycelia with the inoculation of sterilization shovel together with culture medium, transfers to and cultivates in new culture medium, repeats until purifying agaric；Armillaria mellea when in culture medium, vaccination place has just produced shoestring branch, selects wherein to grow vigorous and young tender shoestring, intercepts long 2～3 millimeters with inoculation shovel and moves on slant mediums, cultivates under 25 DEG C of constant temperature, and shoestring is purification strain after covering with culture medium.
3, Germination Strain, Armillaria mellea second class inoculum (original seed) are cultivated
(1) Germination Strain second class inoculum is cultivated: medium component: the wood flour 70% of broad leaf tree, wheat bran or Testa oryzae 26%, sucrose 1%, Gypsum Fibrosum powder 1%, potassium dihydrogen phosphate 1.5%, the magnesium sulfate 0.5% such as green grass or young crops, birch, adds water appropriate.Above composition is fully mixed thoroughly, pinches culture medium with hands and can drip for spending, then bottle, beat the circular hole of diameter 1 centimetre at bottle center, as deep as culture medium 2/3 place.Build bottle cap, autoclaving (1.5 thousand grams/cm) 1 hour, inoculate after cooling.In transfer room, by the mycelia female kind together with about Semen Viciae fabae size, be connected in the circular hole of culture medium, one female plant can connect former in about 5 bottles, build the rearmounted 25 DEG C of culturing room of bottle cap and cultivate.
(2) Armillaria mellea second class inoculum cultivation: medium component is broad leaf tree wood sawdust 3 parts, 1 part of Testa Tritici and twig section, water is appropriate.During making, first the branch of thick fingers is cut into the segment of long 1.5 centimetres, water soaks fully water suction in 12 hours, then mix thoroughly with wood sawdust, Testa Tritici, add water drenched, load in 500 milliliters of wide mouthed bottles, often bottled enter branch section 50～60 sections, be advisable with the 4/5 of bottle capacity, add water a small amount of, autoclaving (1.5 thousand grams/cm) 1 hour after lid bottle cap, after cooling, access mother and plant, be placed in 25 DEG C of constant temperature culture 30 days～45 days.
4, Germination Strain, Armillaria mellea three-class strain (cultigen) are cultivated
(1) Germination Strain three-class strain is cultivated: taking Fagaceae fallen leaves 70%, wood flour 10%, Testa Tritici 15%, magnesium sulfate 0.5%, potassium dihydrogen phosphate 1.5%, carbamide 1%, sucrose 1%, Gypsum Fibrosum 1%, water is appropriate.Bottle after mixing thoroughly, autoclaving (1.5 thousand grams/cm) 1 hour, accesses second class inoculum, builds bottle cap, move into culturing room and cultivate after cooling.
(2) three grades of kinds of Armillaria mellea are cultivated: compost is identical with second class inoculum cultivation with cultural method.After autoclaving cools down, every bottle graft enters two grades of branch strain 1-2 sections, and 25 DEG C of constant temperature culture 30 days～45 days, Armillaria mellea silk and shoestring can cover with full bottle, can be used for cultivating bacterium branch and bacterium material.
5, sowing
(1) bacterium leaf seed dressing: 1. first by three grades of kinds of Germination Strain before sowing, by every square metre of consumption 4 bottles, takes out from seed bottle with the cleek of cleaning, puts in the seed dressing basin of cleaning, bacterium leaf is torn into individual；2. the tender fruit of Rhizoma Gastrodiae gathered and by every square metre, dehiscent fruit is sowed capsule 30～40, shakes out seed；3. seed is loaded planter box and is sprinkling upon on bacterium leaf, simultaneously with the dynamic bacterium leaf of handspring, seed is uniformly mixed on bacterium leaf, and is divided into two parts；
(2) bacterium sowing in bed kind: excavate bacterium bed during sowing, take out bacterium material, at the bottom of raking bed, the first wet fallen leaves of paving a thin layer, then the bacterium leaf divided is sprinkling upon on fallen leaves with 1 part, in statu quo set lower floor's bacterium material, 3～4 cm distance are stayed between bacterium material, blinding is put down to bacterium material, repave wet fallen leaves, spread another part of seed dressing bacterium leaf, put earthing 5-6 centimetre after bacterium material, bed top lid layer leaves moisturizing, then, seeding layer spreads one layer of cultured Armillaria mellea three-class strain, use 8-12 weight portion for every square metre, spread last layer clear water again and soak drenched broad-leaved leaves, every 40-60 weight parts water adds ground phosphate rock 0.3-0.6 weight portion, the upper fine sand covering 8-12cm or fertile soil, moisturizing.
Claims (1)
1. a gastrodin cultivation method, it is characterised in that concrete technical scheme is:
(1) selection and the collection of material are separated:
1. Armillaria mellea separates material: in rainy season in late fall, when Armillaria mellea sporophore is unearthed, and fresh, the form of gathering is complete, healthy and strong, cause harm without disease pest, will the word entity of maturation.Gastrodia Elata Stem Tuber: can gather, in 4-6 month, the Gastrodia Elata Stem Tuber that raw Armillaria mellea is more.2. Germination Strain separation material: the protocorm after collecting tuber of elevated gastrodia germination, the root in orchid, inducing culture Germination Strain sporophore.
(2) isolation technics
Separation method: by Rhizomorph of Armillaria mellea or Gastrodia Elata Stem Tuber, Rhizoma Gastrodiae protocorm, the root of orchid washes surface earth respectively, then cuts the part tissue of needs, with aseptic water washing 3 times, puts into 0.1% mercuric chloride solution and soaks 1 minute；Use aseptic water washing 3 times again, wash away the medicinal liquid of remaining, be placed in sterile petri dish.The materials such as shoestring are cut into respectively segment or the fritter of 1 centimeter length, leaching a moment in penicillin liquid (20 mcg/ml), surface attachment water liquid is sucked with sterilizing filter paper, then it is inoculated on slant medium, put 25 DEG C of constant temperature culture, about 3～7 days, start to grow white hypha and shoestring respectively；During segregant entity, after aseptic water washing sterilization remaining medicinal liquid, exhaust the globule with sterilizing filter paper, then from cap, longitudinally cut entity open with dissecting knife, take its block organization of 0.5 centimetre at stem and cap intersection and be placed in PDA culture medium and cultivate.
Purifying agaric is cultivated: the mycelium that Germination Strain selection colony growth rate is more consistent, cuts mycelia with the inoculation of sterilization shovel together with culture medium, transfers to and cultivates in new culture medium, repeats until purifying agaric；Armillaria mellea when in culture medium, vaccination place has just produced shoestring branch, selects wherein to grow vigorous and young tender shoestring, intercepts long 2～3 millimeters with inoculation shovel and moves on slant mediums, cultivates under 25 DEG C of constant temperature, and shoestring is purification strain after covering with culture medium.
(3) Germination Strain, Armillaria mellea second class inoculum (original seed) are cultivated
Germination Strain second class inoculum is cultivated: medium component: the wood flour 70% of broad leaf tree, wheat bran or Testa oryzae 26%, sucrose 1%, Gypsum Fibrosum powder 1%, potassium dihydrogen phosphate 1.5%, the magnesium sulfate 0.5% such as green grass or young crops, birch, adds water appropriate.Above composition is fully mixed thoroughly, pinches culture medium with hands and can drip for spending, then bottle, beat the circular hole of diameter 1 centimetre at bottle center, as deep as culture medium 2/3 place.Build bottle cap, autoclaving (1.5 thousand grams/cm) 1 hour, inoculate after cooling.In transfer room, by the mycelia female kind together with about Semen Viciae fabae size, be connected in the circular hole of culture medium, one female plant can connect former in about 5 bottles, build the rearmounted 25 DEG C of culturing room of bottle cap and cultivate.
Armillaria mellea second class inoculum cultivation: medium component is broad leaf tree wood sawdust 3 parts, 1 part of Testa Tritici and twig section, water is appropriate.During making, first the branch of thick fingers is cut into the segment of long 1.5 centimetres, water soaks fully water suction in 12 hours, then mix thoroughly with wood sawdust, Testa Tritici, add water drenched, load in 500 milliliters of wide mouthed bottles, often bottled enter branch section 50～60 sections, be advisable with the 4/5 of bottle capacity, add water a small amount of, autoclaving (1.5 thousand grams/cm) 1 hour after lid bottle cap, after cooling, access mother and plant, be placed in 25 DEG C of constant temperature culture 30 days～45 days.
(4) Germination Strain, Armillaria mellea three-class strain (cultigen) are cultivated
Germination Strain three-class strain is cultivated: taking Fagaceae fallen leaves 70%, wood flour 10%, Testa Tritici 15%, magnesium sulfate 0.5%, potassium dihydrogen phosphate 1.5%, carbamide 1%, sucrose 1%, Gypsum Fibrosum 1%, water is appropriate.Bottle after mixing thoroughly, autoclaving (1.5 thousand grams/cm) 1 hour, accesses second class inoculum, builds bottle cap, move into culturing room and cultivate after cooling.
Three grades of kinds of Armillaria mellea are cultivated: compost is identical with second class inoculum cultivation with cultural method.After autoclaving cools down, every bottle graft enters two grades of branch strain 1-2 sections, and 25 DEG C of constant temperature culture 30 days～45 days, Armillaria mellea silk and shoestring can cover with full bottle, can be used for cultivating bacterium branch and bacterium material.
(5) sowing
Bacterium leaf is dressed seed: 1. first by three grades of kinds of Germination Strain before sowing, by every square metre of consumption 4 bottles, takes out with the cleek of cleaning, put in the seed dressing basin of cleaning, bacterium leaf is torn into individual from seed bottle；2. the tender fruit of Rhizoma Gastrodiae gathered and by every square metre, dehiscent fruit is sowed capsule 30～40, shakes out seed；3. seed is loaded planter box and is sprinkling upon on bacterium leaf, simultaneously with the dynamic bacterium leaf of handspring, seed is uniformly mixed on bacterium leaf, and is divided into two parts；
Bacterium sowing in bed kind: excavate bacterium bed during sowing, take out bacterium material, at the bottom of raking bed, the first wet fallen leaves of paving a thin layer, then the bacterium leaf divided is sprinkling upon on fallen leaves with 1 part, in statu quo set lower floor's bacterium material, 3～4 cm distance are stayed between bacterium material, blinding is put down to bacterium material, repave wet fallen leaves, spread another part of seed dressing bacterium leaf, put earthing 5-6 centimetre after bacterium material, bed top lid layer leaves moisturizing, then, seeding layer spreads one layer of cultured Armillaria mellea three-class strain, use 8-12 weight portion for every square metre, spread last layer clear water again and soak drenched broad-leaved leaves, every 40-60 weight parts water adds ground phosphate rock 0.3-0.6 weight portion, the upper fine sand covering 8-12cm or fertile soil, moisturizing.
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