CN105732161A - Preparation method of high-lignin straw decomposition agent - Google Patents
Preparation method of high-lignin straw decomposition agent Download PDFInfo
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- CN105732161A CN105732161A CN201610198008.2A CN201610198008A CN105732161A CN 105732161 A CN105732161 A CN 105732161A CN 201610198008 A CN201610198008 A CN 201610198008A CN 105732161 A CN105732161 A CN 105732161A
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Abstract
The invention relates to a preparation method of a high-lignin straw decomposition agent, belonging to the technical field of straw decomposition agent preparation. The preparation method comprises the following steps: collecting the intestinal juice of worker termite, obtaining the intestinal bacterial colony of termite and culturing; inoculating a domestication medium with the intestinal bacterial colony of termite and the prepared straw decomposition bacterial liquid; screening out the qualified medium bacterial colony and further domesticizing to obtain a lignin straw medium mixed bacterial colony; and inoculating the mixed medium with the mixed bacterial colony and culturing to obtain the decomposition agent. In the invention, the prepared straw decomposition agent is not influenced by the strain formula and contains a large quantity of lignin decomposers, thus the decomposition effect is good, and the application is relatively stable; and moreover, the decomposition time can be greatly shortened, and the straw can be completely decomposed in 6-8 days.
Description
Technical field
The preparation method that the present invention relates to a kind of high lignin straw decomposing inoculant, belongs to straw decomposing inoculant preparing technical field.
Background technology
Straw decomposing inoculant can make the organic waste quick compostings such as straw, organic matter contained in straw and the element such as phosphorus, potassium is made to become the nutrition needed for plant growing, and produce a large amount of beneficial microbe, stimulate crop production, improve the soil organism, strengthen stress resistance of plant, reduce fertilizer application amount, improve crop quality, it is achieved the sustainable development of agricultural.
At present, known straw decomposing inoculant composite by the effective viable bacteria (antibacterial, fungus or actinomycetes) possessing Straw decomposing ability and add carrier prepare, improving material can be played to a certain extent become thoroughly decomposed accumulated temperature, accelerate the effect of the softening of straw material, decomposition, but join owing to being subject to strain group, put forth effort not particularly in lignin fungi selection aspect, in addition the restriction of culture propagation ability and area surroundings difference etc., straw decomposing effect is not ideal enough, and result of use is unstable.
Summary of the invention
The technical problem to be solved: join owing to being subject to strain group for straw decomposing inoculant that is composite with effective viable bacteria and that add carrier prepared, put forth effort not particularly in lignin fungi selection aspect, make its effect of becoming thoroughly decomposed undesirable, the problem that result of use is unstable, provide a kind of intestinal liquid first collecting Coptotermes formosanus Shtrari. worker ant, obtain termite gut bacterium colony and it is cultivated, again it is seeded to domestication culture medium with the straw corrosion bacterium solution prepared, screen qualified culture medium bacterium colony, and carry out continuing domestication to it, obtain high lignin straw medium mixing bacterium colony, finally it is seeded on mixed culture medium and carries out cultivating the method that can prepare decomposing agent.Containing a large amount of lignin fungis in straw decomposing inoculant prepared by the present invention, the impact do not joined by strain group, become thoroughly decomposed effective, use more stable.
For solving above-mentioned technical problem, the present invention adopts the technical scheme as described below to be:
(1) Coptotermes formosanus Shtrari. worker ant is chosen, with straw, it is raised 10～15 days, subsequently with mass concentration be 65% alcoholic solution clean Coptotermes formosanus Shtrari. body surface, wash 2～3 times with sterilized water, again with sterilizing tip tweezers pull-out intestinal, tear intestinal scrotiform stomach, collect intestinal juice and be placed in the sterile saline that mass concentration is 0.9%, by stirring rod, intestinal tissue is smashed to pieces, be prepared into termite gut bacterium mixed liquor；
(2) count by weight, weigh 25～45 parts of clean bean sprout, 25～30 parts of sucrose, 30～45 parts of agar respectively, stirring is mixed with to obtain culture medium solid, subsequently by solid-to-liquid ratio 1:5, is mixed with deionized water and stirring by culture medium solid, sterilization treatment 25～30min at 120 DEG C, it is prepared into culture medium, subsequently the termite gut bacterium mixed liquor of above-mentioned preparation is seeded in culture medium, 25～28 DEG C, shaking table shaken cultivation 3～5 days under 120r/min, it is prepared into termite gut bacterium colony, standby；
(3) straw cleaned and pulverize, 25～30 DEG C of lower seal fermentation process 15～30 days, after its initial fermentation, 1:10 in mass ratio, stalk powder mix with physiological saline solution stirring, filters subsequently and collect filtrate, be prepared into straw and corrode bacterium solution；
(4) count by weight, weigh 35～55 parts of deionized waters, 15～20 parts of agar, 10～15 portions of skim milks and 20～30 parts of stalk powder stirring mixing respectively, sterilization treatment 25～30min at 120 DEG C, it is prepared into domestication culture medium, choose 5 domestication culture medium subsequently, respectively straw is corroded bacterium solution and termite gut colony inoculation and tames culture medium to 5, be placed on 25～28 DEG C, shaking table shaken cultivation 3～5 days under 120r/min；
(5) the culture medium bacterium colony that in domestication culture medium, bacterium colony is evenly distributed and survival rate is higher is observed, it is seeded in another domestication culture medium, control external condition constant, skim milk is reduced every time 5 parts, stalk powder increases by 5 parts, repeat domestication to be cultured to culture medium to be entirely stalk powder be after carbon source, continue cultivation 3～5 generation, be prepared into high lignin straw medium mixing bacterium colony；
(6) count by weight, choose 45～55 portions of Semen Glyciness, 10～15 parts of Semen Maydis powder and 35～40 wheat brans respectively, pulverized and stirred mixing 15～20min, it is prepared into mixed culture medium, regulating water content is 55～60%, subsequently by inoculum concentration 5%, the high lignin straw medium mixing bacterium colony of above-mentioned preparation is seeded on mixed culture medium, after having inoculated, it is placed under 23～25 DEG C of gnotobasiss and cultivates 8～10 days, after cultivation completes, being placed on shady and cool ventilation place, natural air drying can be prepared into a kind of high lignin straw decomposing inoculant.
The application process of the present invention: after crops are gathered in, harrow division of cells with Niu Yili mono-, minizone is built ridge and wraps up dry rice straw also field, every community amount 0.36～0.45kg/m of being separated by with thin film2, then to spreading fertilizer over the fields decomposing agent and the carbamide of above-mentioned preparation respectively, the decomposing agent amount of spreading fertilizer over the fields is 30～33g/m2, carbamide is 70～75g/m2, after having spread fertilizer over the fields, in machinery depression bar to soil, and fill deep water waterlogged plot 4～5 days.After testing, compared with carrying out straw-returning with the decomposing agent not utilizing the present invention to prepare, straw decomposing inoculant prepared by the present invention becomes thoroughly decomposed effective, improves 15～25%, and only can need to become thoroughly decomposed in 6～8 days.
The present invention is compared with additive method, and Advantageous Effects is:
(1) impact that straw decomposing inoculant prepared by the present invention is not joined by strain group, and containing a large amount of lignin fungis so that it becomes thoroughly decomposed effective, uses more stable；
(2) straw decomposing inoculant prepared by the present invention is substantially shorter the time of becoming thoroughly decomposed, and straw can become thoroughly decomposed in 6～8 days；
(3) preparation process is simple, and required cost is low.
Detailed description of the invention
First Coptotermes formosanus Shtrari. worker ant is chosen, with straw, it is raised 10～15 days, subsequently with mass concentration be 65% alcoholic solution clean Coptotermes formosanus Shtrari. body surface, wash 2～3 times with sterilized water, again with sterilizing tip tweezers pull-out intestinal, tear intestinal scrotiform stomach, collect intestinal juice and be placed in the sterile saline that mass concentration is 0.9%, by stirring rod, intestinal tissue is smashed to pieces, be prepared into termite gut bacterium mixed liquor；Count by weight, weigh 25～45 parts of clean bean sprout, 25～30 parts of sucrose, 30～45 parts of agar respectively, stirring is mixed with to obtain culture medium solid, subsequently by solid-to-liquid ratio 1:5, is mixed with deionized water and stirring by culture medium solid, sterilization treatment 25～30min at 120 DEG C, it is prepared into culture medium, subsequently the termite gut bacterium mixed liquor of above-mentioned preparation is seeded in culture medium, 25～28 DEG C, shaking table shaken cultivation 3～5 days under 120r/min, it is prepared into termite gut bacterium colony, standby；Again straw cleaned and pulverize, 25～30 DEG C of lower seal fermentation process 15～30 days, after its initial fermentation, 1:10 in mass ratio, stalk powder mix with physiological saline solution stirring, filters subsequently and collect filtrate, be prepared into straw and corrode bacterium solution；Then count by weight, weigh 35～55 parts of deionized waters, 15～20 parts of agar, 10～15 portions of skim milks and 20～30 parts of stalk powder stirring mixing respectively, sterilization treatment 25～30min at 120 DEG C, it is prepared into domestication culture medium, choose 5 domestication culture medium subsequently, respectively straw is corroded bacterium solution and termite gut colony inoculation and tames culture medium to 5, be placed on 25～28 DEG C, shaking table shaken cultivation 3～5 days under 120r/min；Observe the culture medium bacterium colony that in domestication culture medium, bacterium colony is evenly distributed and survival rate is higher, it is seeded in another domestication culture medium, control external condition constant, skim milk is reduced every time 5 parts, stalk powder increases by 5 parts, repeat domestication to be cultured to culture medium to be entirely stalk powder be after carbon source, continue cultivation 3～5 generation, be prepared into high lignin straw medium mixing bacterium colony；Finally count by weight, choose 45～55 portions of Semen Glyciness, 10～15 parts of Semen Maydis powder and 35～40 wheat brans respectively, pulverized and stirred mixing 15～20min, it is prepared into mixed culture medium, regulating water content is 55～60%, subsequently by inoculum concentration 5%, the high lignin straw medium mixing bacterium colony of above-mentioned preparation is seeded on mixed culture medium, after having inoculated, it is placed under 23～25 DEG C of gnotobasiss and cultivates 8～10 days, after cultivation completes, being placed on shady and cool ventilation place, natural air drying can be prepared into a kind of high lignin straw decomposing inoculant.
Example 1
First Coptotermes formosanus Shtrari. worker ant is chosen, with straw, it is raised 15 days, subsequently with mass concentration be 65% alcoholic solution clean Coptotermes formosanus Shtrari. body surface, wash 3 times with sterilized water, again with sterilizing tip tweezers pull-out intestinal, tear intestinal scrotiform stomach, collect intestinal juice and be placed in the sterile saline that mass concentration is 0.9%, by stirring rod, intestinal tissue is smashed to pieces, be prepared into termite gut bacterium mixed liquor；Count by weight, weigh 45 parts of clean bean sprout, 25 parts of sucrose, 30 parts of agar respectively, stirring is mixed with to obtain culture medium solid, subsequently by solid-to-liquid ratio 1:5, is mixed with deionized water and stirring by culture medium solid, sterilization treatment 30min at 120 DEG C, it is prepared into culture medium, subsequently the termite gut bacterium mixed liquor of above-mentioned preparation is seeded in culture medium, 28 DEG C, shaking table shaken cultivation 5 days under 120r/min, it is prepared into termite gut bacterium colony, standby；Again straw cleaned and pulverize, 30 DEG C of lower seal fermentation process 30 days, after its initial fermentation, 1:10 in mass ratio, stalk powder mix with physiological saline solution stirring, filters subsequently and collect filtrate, be prepared into straw and corrode bacterium solution；Then count by weight, weigh 55 parts of deionized waters, 15 parts of agar, 10 portions of skim milks and 20 parts of stalk powder stirring mixing respectively, sterilization treatment 30min at 120 DEG C, it is prepared into domestication culture medium, choose 5 domestication culture medium subsequently, respectively straw is corroded bacterium solution and termite gut colony inoculation and tames culture medium to 5, be placed on 28 DEG C, shaking table shaken cultivation 5 days under 120r/min；Observe the culture medium bacterium colony that in domestication culture medium, bacterium colony is evenly distributed and survival rate is higher, it is seeded in another domestication culture medium, control external condition constant, skim milk is reduced every time 5 parts, stalk powder increases by 5 parts, repeat domestication to be cultured to culture medium to be entirely stalk powder be after carbon source, continue to cultivate for 5 generations, be prepared into high lignin straw medium mixing bacterium colony；Finally count by weight, choose 55 portions of Semen Glyciness, 10 parts of Semen Maydis powder and 35 wheat brans respectively, pulverized and stir mixing 15min, being prepared into mixed culture medium, regulating water content is 60%, subsequently by inoculum concentration 5%, the high lignin straw medium mixing bacterium colony of above-mentioned preparation is seeded on mixed culture medium, after having inoculated, it is placed under 25 DEG C of gnotobasiss and cultivates 10 days, after cultivation completes, being placed on shady and cool ventilation place, natural air drying can be prepared into a kind of high lignin straw decomposing inoculant.
After crops are gathered in, harrowing division of cells with Niu Yili mono-, minizone is built ridge and wraps up dry rice straw also field, every community amount 0.45kg/m of being separated by with thin film2, then to spreading fertilizer over the fields decomposing agent and the carbamide of above-mentioned preparation respectively, the decomposing agent amount of spreading fertilizer over the fields is 33g/m2, carbamide is 75g/m2, after having spread fertilizer over the fields, in machinery depression bar to soil, and fill deep water waterlogged plot 5 days.After testing, compared with carrying out straw-returning with the decomposing agent not utilizing the present invention to prepare, straw decomposing inoculant prepared by the present invention becomes thoroughly decomposed effective, improves 25%, and only can need to become thoroughly decomposed in 8 days.
Example 2
First Coptotermes formosanus Shtrari. worker ant is chosen, with straw, it is raised 10 days, subsequently with mass concentration be 65% alcoholic solution clean Coptotermes formosanus Shtrari. body surface, wash 2 times with sterilized water, again with sterilizing tip tweezers pull-out intestinal, tear intestinal scrotiform stomach, collect intestinal juice and be placed in the sterile saline that mass concentration is 0.9%, by stirring rod, intestinal tissue is smashed to pieces, be prepared into termite gut bacterium mixed liquor；Count by weight, weigh 25 parts of clean bean sprout, 30 parts of sucrose, 45 parts of agar respectively, stirring is mixed with to obtain culture medium solid, subsequently by solid-to-liquid ratio 1:5, is mixed with deionized water and stirring by culture medium solid, sterilization treatment 25min at 120 DEG C, it is prepared into culture medium, subsequently the termite gut bacterium mixed liquor of above-mentioned preparation is seeded in culture medium, 25 DEG C, shaking table shaken cultivation 3 days under 120r/min, it is prepared into termite gut bacterium colony, standby；Again straw cleaned and pulverize, 25 DEG C of lower seal fermentation process 15 days, after its initial fermentation, 1:10 in mass ratio, stalk powder mix with physiological saline solution stirring, filters subsequently and collect filtrate, be prepared into straw and corrode bacterium solution；Then count by weight, weigh 35 parts of deionized waters, 20 parts of agar, 15 portions of skim milks and 30 parts of stalk powder stirring mixing respectively, sterilization treatment 25min at 120 DEG C, it is prepared into domestication culture medium, choose 5 domestication culture medium subsequently, respectively straw is corroded bacterium solution and termite gut colony inoculation and tames culture medium to 5, be placed on 25 DEG C, shaking table shaken cultivation 3 days under 120r/min；Observe the culture medium bacterium colony that in domestication culture medium, bacterium colony is evenly distributed and survival rate is higher, it is seeded in another domestication culture medium, control external condition constant, skim milk is reduced every time 5 parts, stalk powder increases by 5 parts, repeat domestication to be cultured to culture medium to be entirely stalk powder be after carbon source, continue to cultivate for 3 generations, be prepared into high lignin straw medium mixing bacterium colony；Finally count by weight, choose 45 portions of Semen Glyciness, 15 parts of Semen Maydis powder and 40 wheat brans respectively, pulverized and stir mixing 15min, being prepared into mixed culture medium, regulating water content is 55%, subsequently by inoculum concentration 5%, the high lignin straw medium mixing bacterium colony of above-mentioned preparation is seeded on mixed culture medium, after having inoculated, it is placed under 23 DEG C of gnotobasiss and cultivates 8 days, after cultivation completes, being placed on shady and cool ventilation place, natural air drying can be prepared into a kind of high lignin straw decomposing inoculant.
After crops are gathered in, harrowing division of cells with Niu Yili mono-, minizone is built ridge and wraps up dry rice straw also field, every community amount 0.36kg/m of being separated by with thin film2, then to spreading fertilizer over the fields decomposing agent and the carbamide of above-mentioned preparation respectively, the decomposing agent amount of spreading fertilizer over the fields is 30g/m2, carbamide is 70g/m2, after having spread fertilizer over the fields, in machinery depression bar to soil, and fill deep water waterlogged plot 4 days.After testing, compared with carrying out straw-returning with the decomposing agent not utilizing the present invention to prepare, straw decomposing inoculant prepared by the present invention becomes thoroughly decomposed effective, improves 15%, and only can need to become thoroughly decomposed in 6 days.
Example 3
First Coptotermes formosanus Shtrari. worker ant is chosen, with straw, it is raised 12 days, subsequently with mass concentration be 65% alcoholic solution clean Coptotermes formosanus Shtrari. body surface, wash 2 times with sterilized water, again with sterilizing tip tweezers pull-out intestinal, tear intestinal scrotiform stomach, collect intestinal juice and be placed in the sterile saline that mass concentration is 0.9%, by stirring rod, intestinal tissue is smashed to pieces, be prepared into termite gut bacterium mixed liquor；Count by weight, weigh 40 parts of clean bean sprout, 30 parts of sucrose, 30 parts of agar respectively, stirring is mixed with to obtain culture medium solid, subsequently by solid-to-liquid ratio 1:5, is mixed with deionized water and stirring by culture medium solid, sterilization treatment 27min at 120 DEG C, it is prepared into culture medium, subsequently the termite gut bacterium mixed liquor of above-mentioned preparation is seeded in culture medium, 26 DEG C, shaking table shaken cultivation 4 days under 120r/min, it is prepared into termite gut bacterium colony, standby；Again straw cleaned and pulverize, 27 DEG C of lower seal fermentation process 20 days, after its initial fermentation, 1:10 in mass ratio, stalk powder mix with physiological saline solution stirring, filters subsequently and collect filtrate, be prepared into straw and corrode bacterium solution；Then count by weight, weigh 40 parts of deionized waters, 20 parts of agar, 10 portions of skim milks and 30 parts of stalk powder stirring mixing respectively, sterilization treatment 27min at 120 DEG C, it is prepared into domestication culture medium, choose 5 domestication culture medium subsequently, respectively straw is corroded bacterium solution and termite gut colony inoculation and tames culture medium to 5, be placed on 26 DEG C, shaking table shaken cultivation 4 days under 120r/min；Observe the culture medium bacterium colony that in domestication culture medium, bacterium colony is evenly distributed and survival rate is higher, it is seeded in another domestication culture medium, control external condition constant, skim milk is reduced every time 5 parts, stalk powder increases by 5 parts, repeat domestication to be cultured to culture medium to be entirely stalk powder be after carbon source, continue to cultivate for 4 generations, be prepared into high lignin straw medium mixing bacterium colony；Finally count by weight, choose 50 portions of Semen Glyciness, 15 parts of Semen Maydis powder and 35 wheat brans respectively, pulverized and stir mixing 17min, being prepared into mixed culture medium, regulating water content is 57%, subsequently by inoculum concentration 5%, the high lignin straw medium mixing bacterium colony of above-mentioned preparation is seeded on mixed culture medium, after having inoculated, it is placed under 24 DEG C of gnotobasiss and cultivates 9 days, after cultivation completes, being placed on shady and cool ventilation place, natural air drying can be prepared into a kind of high lignin straw decomposing inoculant.
After crops are gathered in, harrowing division of cells with Niu Yili mono-, minizone is built ridge and wraps up dry rice straw also field, every community amount 0.42kg/m of being separated by with thin film2, then to spreading fertilizer over the fields decomposing agent and the carbamide of above-mentioned preparation respectively, the decomposing agent amount of spreading fertilizer over the fields is 32g/m2, carbamide is 72g/m2, after having spread fertilizer over the fields, in machinery depression bar to soil, and fill deep water waterlogged plot 4 days.After testing, compared with carrying out straw-returning with the decomposing agent not utilizing the present invention to prepare, straw decomposing inoculant prepared by the present invention becomes thoroughly decomposed effective, improves 20%, and only can need to become thoroughly decomposed in 7 days.
Claims (1)
1. the preparation method of a high lignin straw decomposing inoculant, it is characterised in that concrete preparation process is:
(1) Coptotermes formosanus Shtrari. worker ant is chosen, with straw, it is raised 10～15 days, subsequently with mass concentration be 65% alcoholic solution clean Coptotermes formosanus Shtrari. body surface, wash 2～3 times with sterilized water, again with sterilizing tip tweezers pull-out intestinal, tear intestinal scrotiform stomach, collect intestinal juice and be placed in the sterile saline that mass concentration is 0.9%, by stirring rod, intestinal tissue is smashed to pieces, be prepared into termite gut bacterium mixed liquor；
(2) count by weight, weigh 25～45 parts of clean bean sprout, 25～30 parts of sucrose, 30～45 parts of agar respectively, stirring is mixed with to obtain culture medium solid, subsequently by solid-to-liquid ratio 1:5, is mixed with deionized water and stirring by culture medium solid, sterilization treatment 25～30min at 120 DEG C, it is prepared into culture medium, subsequently the termite gut bacterium mixed liquor of above-mentioned preparation is seeded in culture medium, 25～28 DEG C, shaking table shaken cultivation 3～5 days under 120r/min, it is prepared into termite gut bacterium colony, standby；
(3) straw cleaned and pulverize, 25～30 DEG C of lower seal fermentation process 15～30 days, after its initial fermentation, 1:10 in mass ratio, stalk powder mix with physiological saline solution stirring, filters subsequently and collect filtrate, be prepared into straw and corrode bacterium solution；
(4) count by weight, weigh 35～55 parts of deionized waters, 15～20 parts of agar, 10～15 portions of skim milks and 20～30 parts of stalk powder stirring mixing respectively, sterilization treatment 25～30min at 120 DEG C, it is prepared into domestication culture medium, choose 5 domestication culture medium subsequently, respectively straw is corroded bacterium solution and termite gut colony inoculation and tames culture medium to 5, be placed on 25～28 DEG C, shaking table shaken cultivation 3～5 days under 120r/min；
(5) the culture medium bacterium colony that in domestication culture medium, bacterium colony is evenly distributed and survival rate is higher is observed, inoculated in another domestication culture medium, control external condition constant, skim milk is reduced every time 5 parts, stalk powder increases by 5 parts, repeat domestication to be cultured to culture medium to be entirely stalk powder be after carbon source, continue cultivation 3～5 generation, be prepared into high lignin straw medium mixing bacterium colony；
(6) count by weight, choose 45～55 portions of Semen Glyciness, 10～15 parts of Semen Maydis powder and 35～40 wheat brans respectively, pulverized and stirred mixing 15～20min, it is prepared into mixed culture medium, regulating water content is 55～60%, subsequently by inoculum concentration 5%, the high lignin straw medium mixing bacterium colony of above-mentioned preparation is seeded on mixed culture medium, after having inoculated, it is placed under 23～25 DEG C of gnotobasiss and cultivates 8～10 days, after cultivation completes, being placed on shady and cool ventilation place, natural air drying can be prepared into a kind of high lignin straw decomposing inoculant.
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| CN201610198008.2A CN105732161A (en) | 2016-03-31 | 2016-03-31 | Preparation method of high-lignin straw decomposition agent |
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN106495769A (en) * | 2016-10-11 | 2017-03-15 | 常州市鼎升环保科技有限公司 | A kind of preparation method of straw decomposing inoculant |
| CN110771426A (en) * | 2019-10-21 | 2020-02-11 | 郭红伟 | Stable culture method of phellinus igniarius strains |
-
2016
- 2016-03-31 CN CN201610198008.2A patent/CN105732161A/en not_active Withdrawn
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN106495769A (en) * | 2016-10-11 | 2017-03-15 | 常州市鼎升环保科技有限公司 | A kind of preparation method of straw decomposing inoculant |
| CN110771426A (en) * | 2019-10-21 | 2020-02-11 | 郭红伟 | Stable culture method of phellinus igniarius strains |
| CN110771426B (en) * | 2019-10-21 | 2021-08-27 | 郭红伟 | Stable culture method of phellinus igniarius strains |
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