CN103688753A - Solid-liquid alternating culturing method for mycelia of amanita rubrovolvata imai - Google Patents
Solid-liquid alternating culturing method for mycelia of amanita rubrovolvata imai Download PDFInfo
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Abstract
The invention relates to a solid-liquid alternating culturing method for mycelia of amanita rubrovolvata imai and belongs to the technical field of culturing of macro fungi. The solid-liquid alternating culturing method for mycelia of amanita rubrovolvata imai is characterized by comprising the steps of: performing solid culturing on the induced and propagated mycelia for 4-6 generations and then performing the liquid culturing on the induced and propagated mycelia for 3-4 generations by using the solid-liquid alternating culturing method; then performing the solid culturing, wherein the growth speed of the mycelia is 8-10 times that of the mycelia before the liquid culturing, and the growth is speeded obviously. According to the invention, the propagation speed of the mycelia is speeded up by simply changing the culturing method, the operation is simple and easy to implement, the effect is obvious, and the production cost is low. The toxin of the amanita has potential application in the field of developing new specific medicines such as antineoplastic medicines, antibiosis and antiviral medicines, sedatives or narcotics, but is different to develop and apply due to the bottleneck problems such as the valuable and rare resource and the difficulty in artificial acclimatization so far. According to the invention, focused research is performed on the problems for limiting the pure culturing such as very slow growth of the mycelia of amanita rubrovolvata imai, and conditions are provided to the large-scale culturing of the mycelia, the artificial acclimatization cultivation in future and the like.
Description
Technical field
A kind of method that the present invention relates to red holder Amanita fuliginea filament solid-liquid alternate culture, belongs to biological technical field, specifically belongs to the indoor cultivation category of poisonous macro fungi.
Background technology
Amanita (
amanita) be under the jurisdiction of Basidiomycotina, Hymenomycetes, Agaricales, Amanitaceae (
amanitaceae), be in poisonous macro fungi more special, more valuable one worldwidely blazon large genus, its species diversity is enriched very much.The whole world has been reported nearly 400 kinds, nearly 100 kinds (containing subspecies, mutation and the modification) that China has recorded.According to textual criticism, China still has many kinds not yet study and name at present.But nowadays, along with the appearance of global ecological crisis, many Macro-Fungi Resources of China have been accused danger, in slump of disastrous proportions and Critical Condition, and part plant be faced with may be also not by human knowledge or while finding that it is worth, the risk of just having become extinct.
Major part kind in Amanita belongs to famous hypertoxic bacterium, it is reported, and because eating the wild gill fungus bacterium person of being poisoned to death 95% by mistake, be because of due to Amanita fuliginea.Scientists is real interested to Amanita fuliginea is its toxicity, and therefore, before 140 years, people have just started the research of In The Studies On Toxins of Amanita greatly.The application study of amatoxin is mainly reflected in following several respects: 1. can be used for studying expression, regulation and control and the celluar localization of eukaryotic gene; 2. can develop antibacterial, antiviral special efficacy new drug; 3. can antitumor medicine screening; 4. can develop calmness, anesthesia and other specific drug; 5. can be used for biological anti-smelting etc.
The Main Bottleneck problem that causes at present goose cream to be difficult to exploitation and sustainability utilization has: 1. goose cream resource is rare, an its kind of group of mean people, individual few, yield poorly, distributed quantity is extremely limited, in addition responsive to habitat, once habitat is damaged, very easily disappear or extinction, belong to the special danger biological group that causes, this has increased its further studied, difficulty of utilizing; 2. Amanita, belongs to Applying Ectomycorrhizal Fungi mostly, and its most absolutely kind still belongs to the difficult microorganism of cultivating or failing to cultivate of occurring in nature, is difficult to artificial, semi-artificial cultivation.Therefore, also there is no both at home and abroad so far a kind of toxin product that can scale exploitation, the existing Peptide toxin as biochemical reagents is expensive (before more than 10 year every gram at 100,000 dollars of left and right-Chen Zuohong etc., 1999) still, is difficult to meet scientific research and application is required.
The pure culture of goose cream is prerequisite or the key that guarantees that goose cream sustainability of natural resources utilizes; but this still belongs to the problem that is difficult to capture at present; have many blind spots, wherein mycelia is difficult to induction, mycelial growth is very slowly restriction and important step or the factor that affects pure culture.
Red holder goose cream (
a.rubrovolvata S. Imai), belong to goose cream subgenus (Amanita subgen. Amanita) goose cream group (sect. Amanita), its population distributes less, successfully be separated at present the mycelia of this kind, but the growth of mycelia is still very slow early stage, is difficult to expand numerous, the present invention levies this problem, change cultural method, greatly improved the growth rate of mycelia, for important foundation has been established in the research and development utilization of amatoxin etc.
Summary of the invention
The object of the invention is to, provide a kind of production cost low, easily realize, can make the cultural method of Amanita fuliginea silk Fast Growth.
Technical assignment of the present invention is, adopt solid-liquid alternate culture method, to accelerate mycelial reproduction speed, described in it, feature is as follows: will induce and expand numerous mycelium, solid culture is after 4～6 generations, 3～4 generations of liquid culture, then carry out 2～3 generations of solid culture, adopt the cultural method of solid-liquid-solid;
Induction and the expanding propagation method of described red holder Amanita fuliginea filament are as follows: the strong shape of growth, the tender fruit body of the children of parachute-opening are not plucked in field; Get the about 0.3cm of inside sterile tissue piece of cap and stem junction
2size, embeds inducing culture gently, and described medium component is: potato 200g/L, glucose 13～15g/L, yeast extract 1.20～1.40g/L, MgSO
41.00g/L, CaCl
21.00g/L, KH
2pO
4brewer's wort 120～the 140ml of 0.50g/L, 16.0 Baumes, agar 10.00g/L, controlling medium pH value is 5.8～6.0, dark cultivation 46～55 days at 22～23 ℃, tissue block surface grows fine hair shape white hypha; Mycelia subculture in above-mentioned inducing culture is expanded numerous 2～3 times;
Described solid culture method is as follows: the diameter that mycelium is proceeded to containing solid culture medium is in 9cm culture dish, dark cultivation 70～80 days at 22～23 ℃, and when mycelia grows to 3.5～4.5cm, switching, cultivated for 3～6 generations; Described solid culture based component is: potato 200g/L, glucose 15～18g/L, yeast extract 1.20～1.40g/L, ZnSO
40.40～0.60 g/L, MgSO
40.40～0.60g/L, zeatin ZT 1.00～1.20mg/L, complex habitat material 90～100g/L, agar 10.00g/L, controlling medium pH value is 5.8～6.0;
The preparation method of the complex habitat material in described solid culture medium is as follows: at red holder goose cream vegetatively, get Qinggang, Yunnan tree root, Qinggang, Yunnan fallen leaves of growth soil, symbiosis, take back laboratory, dry, with high speed disintegrator, pulverize respectively, in native: the ratio of root: leaf=1:1:1 is mixed;
Described liquid cultivating method is as follows: get in culture dish through the solid culture mycelium in 3～6 generations, the aseptic card punch that is 6mm with diameter, under aseptic condition, cut bacterium sheet, reject medium above, proceed to containing in the 250ml triangular flask of 100ml liquid nutrient medium, 4 of every bottle graft kinds, being placed in rotating speed is 150r/min, temperature is to cultivate on the shaking table of 25 ℃, after 20 days, when mycelia grows up to the tiny bacterium ball of furcella shape of dispersion, getting bacterium liquid that 10ml cultivates moves in the medium of newly joining and carries out subculture, 20～25 days, treat that little bacterium ball becomes large gradually, while being cotton-shaped agglomerate, get final product subculture or go back in solid culture medium, liquid culture based component described in it is: potato 200g/L, glucose 15～18g/L, yeast extract 1.20～1.40g/L, ZnSO
40.40～0.60g/L, MgSO
40.40～0.60g/L, ZT1.00～1.20mg/L, complex habitat material water extraction liquid 400～500ml, controlling medium pH value is 5.8～6.0,
The preparation method of described complex habitat material water extraction liquid is as follows: get above complex habitat material 200g, add 1000ml distilled water immersion after 1 day, 1～2hr is boiled in heating, filters, and filtrate heating is condensed into 400～500ml;
The cotton-shaped mycelium pellet in 3～4 generations of described process liquid culture, then by carrying out above 2～3 generations of solid culture, mycelial growth rate is without 8～10 times before liquid culture, growth is obviously accelerated.
The invention has the beneficial effects as follows: simply change training method, can greatly accelerate mycelial reproduction speed, its feature is as follows:
(1) easy realization simple to operate: by the alternate culture method of solid-liquid-solid, can obviously improve mycelial growth rate;
(2) benefit is obvious: through liquid culture, then proceed to solid culture, mycelial growth rate is without 8～10 times before liquid culture, and growth is obviously accelerated;
(3) production cost is low: the present invention adopts cellar culture mode, and just two kinds of modes hocket; Medium is in conjunction with adding complex habitat material, and this material is taken from field, be easy to get, so the whole incubation of the present invention does not relate to expensive biological or chemical medicine, does not also need special equipment.
Embodiment
Following examples of implementation are to further illustrate of the present invention, are not limitations of the present invention.
example one:
The strong shape of growth, the tender fruit body of the children of parachute-opening are not plucked in field; Get the about 0.3cm of inside sterile tissue piece of cap and stem junction
2size, embeds inducing culture gently, and described medium component is: potato 200g/L, glucose 13g/L, yeast extract 1.20g/L, MgSO
41.00g/L, CaCl
21.00g/L, KH
2pO
4the brewer's wort 120ml of 0.50g/L, 16.0 Baumes, agar 10.00g/L, controlling medium pH value is 5.8, dark cultivation 46 days at 22～23 ℃, tissue block surface grows fine hair shape white hypha; Mycelia subculture in above-mentioned inducing culture is expanded numerous 3 times;
The diameter again mycelium being proceeded to containing solid culture medium is in 9cm culture dish, dark cultivation 70 days at 22～23 ℃, and when mycelia grows to 3.5cm, switching, cultivated for 4 generations; Described solid culture based component is: potato 200g/L, glucose 15g/L, yeast extract 1.20g/L, ZnSO
40.40 g/L, MgSO
40.40g/L, zeatin ZT 1.00mg/L, complex habitat material 90g/L, agar 10.00g/L, controlling medium pH value is 5.8; The preparation method of described complex habitat material is as follows: at red holder goose cream vegetatively, get Qinggang, Yunnan tree root, Qinggang, Yunnan fallen leaves of growth soil, symbiosis, take back laboratory, dry, pulverize respectively, in native: the ratio of root: leaf=1:1:1 is mixed with high speed disintegrator;
Get in culture dish through the solid culture mycelium in 3 generations, the aseptic card punch that is 6mm with diameter, under aseptic condition, cut bacterium sheet, reject medium above, proceed to containing in the 250ml triangular flask of 100ml liquid nutrient medium, 4 of every bottle graft kinds, be placed in rotating speed and be 150r/min, temperature and be on the shaking table of 25 ℃ and cultivate, after 20 days, when mycelia grows up to the tiny bacterium ball of furcella shape of dispersion, get bacterium liquid that 10ml cultivates and move in the medium of newly joining and carry out subculture, 20 days, treat that little bacterium ball becomes greatly gradually, while being cotton-shaped agglomerate, can subculture or go back in solid culture medium; Liquid culture based component described in it is: potato 200g/L, glucose 15g/L, yeast extract 1.20g/L, ZnSO
40.40g/L, MgSO
40.40g/L, ZT1.00mg/L, complex habitat material water extraction liquid 400ml, controlling medium pH value is 5.8; The preparation method of described complex habitat material water extraction liquid is as follows: get above complex habitat material 200g, add 1000ml distilled water immersion after 1 day, 1hr is boiled in heating, filters, and filtrate heating is condensed into 400ml;
By the process liquid culture cotton-shaped mycelium pellet in 3 generations, then cultivated for 2 generations by above solid culture method, mycelial growth rate is without 8 times before liquid culture, and growth is obviously accelerated.
example two:
The strong shape of growth, the tender fruit body of the children of parachute-opening are not plucked in field; Get the about 0.3cm of inside sterile tissue piece of cap and stem junction
2size, embeds inducing culture gently, and described medium component is: potato 200g/L, glucose 15g/L, yeast extract 1.40g/L, MgSO
41.00g/L, CaCl
21.00g/L, KH
2pO
4the brewer's wort 140ml of 0.50g/L, 16.0 Baumes, agar 10.00g/L, controlling medium pH value is 6.0, dark cultivation 55 days at 22～23 ℃, tissue block surface grows fine hair shape white hypha; Mycelia subculture in above-mentioned inducing culture is expanded numerous 2 times;
The diameter again mycelium being proceeded to containing solid culture medium is in 9cm culture dish, dark cultivation 80 days at 22～23 ℃, and when mycelia grows to 4.5cm, switching, cultivated for 6 generations; Described solid culture based component is: potato 200g/L, glucose 18g/L, yeast extract 1.40g/L, ZnSO
40.60 g/L, MgSO
40.60g/L, zeatin ZT1.20mg/L, complex habitat material 100g/L, agar 10.00g/L, controlling medium pH value is 6.0; The preparation method of described complex habitat material is as follows: at red holder goose cream vegetatively, get Qinggang, Yunnan tree root, Qinggang, Yunnan fallen leaves of growth soil, symbiosis, take back laboratory, dry, pulverize respectively, in native: the ratio of root: leaf=1:1:1 is mixed with high speed disintegrator;
Get in culture dish through the solid culture mycelium in 6 generations, the aseptic card punch that is 6mm with diameter, under aseptic condition, cut bacterium sheet, reject medium above, proceed to containing in the 250ml triangular flask of 100ml liquid nutrient medium, 4 of every bottle graft kinds, be placed in rotating speed and be 150r/min, temperature and be on the shaking table of 25 ℃ and cultivate, after 20 days, when mycelia grows up to the tiny bacterium ball of furcella shape of dispersion, get bacterium liquid that 10ml cultivates and move in the medium of newly joining and carry out subculture, 25 days, treat that little bacterium ball becomes greatly gradually, while being cotton-shaped agglomerate, can subculture or go back in solid culture medium; Liquid culture based component described in it is: potato 200g/L, glucose 18g/L, yeast extract 1.40g/L, ZnSO
40.60g/L, MgSO
40.60g/L, ZT1.20mg/L, complex habitat material water extraction liquid 500ml, controlling medium pH value is 6.0; The preparation method of described complex habitat material water extraction liquid is as follows: get above complex habitat material 200g, add 1000ml distilled water immersion after 1 day, 2hr is boiled in heating, filters, and filtrate heating is condensed into 500ml;
By the process liquid culture cotton-shaped mycelium pellet in 4 generations, then cultivated for 3 generations by above solid culture method, mycelial growth rate is without 10 times before liquid culture, and growth is obviously accelerated.
example three:
The strong shape of growth, the tender fruit body of the children of parachute-opening are not plucked in field; Get the about 0.3cm of inside sterile tissue piece of cap and stem junction
2size, embeds inducing culture gently, and described medium component is: potato 200g/L, glucose 14g/L, yeast extract 1.30g/L, MgSO
41.00g/L, CaCl
21.00g/L, KH
2pO
4the brewer's wort 130ml of 0.50g/L, 16.0 Baumes, agar 10.00g/L, controlling medium pH value is 5.8, dark cultivation 50 days at 22～23 ℃, tissue block surface grows fine hair shape white hypha; Mycelia subculture in above-mentioned inducing culture is expanded numerous 3 times;
The diameter again mycelium being proceeded to containing solid culture medium is in 9cm culture dish, dark cultivation 75 days at 22～23 ℃, and when mycelia grows to 4.0cm, switching, cultivated for 5 generations; Described solid culture based component is: potato 200g/L, glucose 17g/L, yeast extract 1.30g/L, ZnSO
40.50 g/L, MgSO
40.50g/L, zeatin ZT 1.10mg/L, complex habitat material 97g/L, agar 10.00g/L, controlling medium pH value is 5.8; The preparation method of described complex habitat material is as follows: at red holder goose cream vegetatively, get Qinggang, Yunnan tree root, Qinggang, Yunnan fallen leaves of growth soil, symbiosis, take back laboratory, dry, pulverize respectively, in native: the ratio of root: leaf=1:1:1 is mixed with high speed disintegrator;
Get in culture dish through the solid culture mycelium in 5 generations, the aseptic card punch that is 6mm with diameter, under aseptic condition, cut bacterium sheet, reject medium above, proceed to containing in the 250ml triangular flask of 100ml liquid nutrient medium, 4 of every bottle graft kinds, be placed in rotating speed and be 150r/min, temperature and be on the shaking table of 25 ℃ and cultivate, after 20 days, when mycelia grows up to the tiny bacterium ball of furcella shape of dispersion, get bacterium liquid that 10ml cultivates and move in the medium of newly joining and carry out subculture, 23 days, treat that little bacterium ball becomes greatly gradually, while being cotton-shaped agglomerate, can subculture or go back in solid culture medium; Liquid culture based component described in it is: potato 200g/L, glucose 17g/L, yeast extract 1.30g/L, ZnSO
40.50g/L, MgSO
40.50g/L, ZT1.15mg/L, complex habitat material water extraction liquid 450ml, controlling medium pH value is 5.8; The preparation method of described complex habitat material water extraction liquid is as follows: get above complex habitat material 200g, add 1000ml distilled water immersion after 1 day, 1.5hr is boiled in heating, filters, and filtrate heating is condensed into 450ml;
By the process liquid culture cotton-shaped mycelium pellet in 3 generations, then cultivated for 3 generations by above solid culture method, mycelial growth rate is without 9 times before liquid culture, and growth is obviously accelerated.
example four:
The strong shape of growth, the tender fruit body of the children of parachute-opening are not plucked in field; Get the about 0.3cm of inside sterile tissue piece of cap and stem junction
2size, embeds inducing culture gently, and described medium component is: potato 200g/L, glucose 13g/L, yeast extract 1.30g/L, MgSO
41.00g/L, CaCl
21.00g/L, KH
2pO
4the brewer's wort 135ml of 0.50g/L, 16.0 Baumes, agar 10.00g/L, controlling medium pH value is 6.0, dark cultivation 53 days at 22～23 ℃, tissue block surface grows fine hair shape white hypha; Mycelia subculture in above-mentioned inducing culture is expanded numerous 2 times;
The diameter again mycelium being proceeded to containing solid culture medium is in 9cm culture dish, dark cultivation 73 days at 22～23 ℃, and when mycelia grows to 3.8cm, switching, cultivated for 3 generations; Described solid culture based component is: potato 200g/L, glucose 16g/L, yeast extract 1.25g/L, ZnSO
40.55 g/L, MgSO
40.55g/L, zeatin ZT1.15mg/L, complex habitat material 94g/L, agar 10.00g/L, controlling medium pH value is 6.0; The preparation method of described complex habitat material is as follows: at red holder goose cream vegetatively, get Qinggang, Yunnan tree root, Qinggang, Yunnan fallen leaves of growth soil, symbiosis, take back laboratory, dry, pulverize respectively, in native: the ratio of root: leaf=1:1:1 is mixed with high speed disintegrator;
Get in culture dish through the solid culture mycelium in 4 generations, the aseptic card punch that is 6mm with diameter, under aseptic condition, cut bacterium sheet, reject medium above, proceed to containing in the 250ml triangular flask of 100ml liquid nutrient medium, 4 of every bottle graft kinds, be placed in rotating speed and be 150r/min, temperature and be on the shaking table of 25 ℃ and cultivate, after 20 days, when mycelia grows up to the tiny bacterium ball of furcella shape of dispersion, get bacterium liquid that 10ml cultivates and move in the medium of newly joining and carry out subculture, 21 days, treat that little bacterium ball becomes greatly gradually, while being cotton-shaped agglomerate, can subculture or go back in solid culture medium; Liquid culture based component described in it is: potato 200g/L, glucose 16g/L, yeast extract 1.30g/L, ZnSO
40.55g/L, MgSO
40.55g/L, ZT1.10mg/L, complex habitat material water extraction liquid 430ml, controlling medium pH value is 6.0; The preparation method of described complex habitat material water extraction liquid is as follows: get above complex habitat material 200g, add 1000ml distilled water immersion after 1 day, 1.2hr is boiled in heating, filters, and filtrate heating is condensed into 430ml;
By the process liquid culture cotton-shaped mycelium pellet in 4 generations, then cultivated for 2 generations by above solid culture method, mycelial growth rate is without 8.5 times before liquid culture, and growth is obviously accelerated.
Claims (7)
1. the method for a red holder Amanita fuliginea filament solid-liquid alternate culture, it is characterized by, adopt solid-liquid alternate culture method, can greatly accelerate mycelial reproduction speed, cultural method described in it is: will induce and expand numerous mycelium, solid culture be after 4～6 generations, 3～4 generations of liquid culture, carry out again 2～3 generations of solid culture, adopt the cultural method of solid-liquid-solid.
2. the method for red holder Amanita fuliginea filament solid-liquid alternate culture according to claim 1, is characterized in that, induction and the expanding propagation method of red holder Amanita fuliginea filament are as follows: the strong shape of growth, the tender fruit body of the children of parachute-opening are not plucked in field; Get the about 0.3cm of inside sterile tissue piece of cap and stem junction
2size, embeds inducing culture gently, and described medium component is: potato 200g/L, glucose 13～15g/L, yeast extract 1.20～1.40g/L, MgSO
41.00g/L, CaCl
21.00g/L, KH
2pO
4brewer's wort 120～the 140ml of 0.50g/L, 16.0 Baumes, agar 10.00g/L, controlling medium pH value is 5.8～6.0, dark cultivation 46～55 days at 22～23 ℃, tissue block surface grows fine hair shape white hypha; Mycelia subculture in above-mentioned inducing culture is expanded numerous 2～3 times.
3. the method for red holder Amanita fuliginea filament solid-liquid alternate culture according to claim 1, it is characterized in that, solid culture method is as follows: the diameter that mycelium is proceeded to containing solid culture medium is in 9cm culture dish, dark cultivation 70～80 days at 22～23 ℃, when mycelia grows to 3.5～4.5cm, switching, cultivated for 3～6 generations; Described solid culture based component is: potato 200g/L, glucose 15～18g/L, yeast extract 1.20～1.40g/L, ZnSO
40.40～0.60 g/L, MgSO
40.40～0.60g/L, zeatin ZT 1.00～1.20mg/L, complex habitat material 90～100g/L, agar 10.00g/L, controlling medium pH value is 5.8～6.0.
4. solid culture method according to claim 3, it is characterized in that, the preparation method of the complex habitat material in described solid culture medium is as follows: at red holder goose cream vegetatively, get Qinggang, Yunnan tree root, Qinggang, Yunnan fallen leaves of growth soil, symbiosis, take back laboratory, dry, with high speed disintegrator, pulverize respectively, in native: the ratio of root: leaf=1:1:1 is mixed.
5. the method for red holder Amanita fuliginea filament solid-liquid alternate culture according to claim 1, it is characterized in that, liquid cultivating method is as follows: get in culture dish through the solid culture mycelium in 3～6 generations, the aseptic card punch that is 6mm with diameter, under aseptic condition, cut bacterium sheet, reject medium above, proceed to containing in the 250ml triangular flask of 100ml liquid nutrient medium, 4 of every bottle graft kinds, being placed in rotating speed is 150r/min, temperature is to cultivate on the shaking table of 25 ℃, after 20 days, when mycelia grows up to the tiny bacterium ball of furcella shape of dispersion, getting bacterium liquid that 10ml cultivates moves in the medium of newly joining and carries out subculture, 20～25 days, treat that little bacterium ball becomes large gradually, while being cotton-shaped agglomerate, get final product subculture or go back in solid culture medium, liquid culture based component described in it is: potato 200g/L, glucose 15～18g/L, yeast extract 1.20～1.40g/L, ZnSO
40.40～0.60g/L, MgSO
40.40～0.60g/L, ZT1.00～1.20mg/L, complex habitat material water extraction liquid 400～500ml, controlling medium pH value is 5.8～6.0.
6. liquid cultivating method according to claim 5, it is characterized in that, the preparation method of complex habitat material water extraction liquid is as follows: get the complex habitat material 200g in claim 4, add 1000ml distilled water immersion after 1 day, 1～2hr is boiled in heating, filter, filtrate heating is condensed into 400～500ml.
7. liquid culture according to claim 5, is characterized in that, through the cotton-shaped mycelium pellet in 3～4 generations of liquid culture, then carries out 2～3 generations of solid culture by claim 3, and mycelial growth rate be without 8～10 times before liquid culture, and growth is quickening obviously.
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103980060A (en) * | 2014-06-05 | 2014-08-13 | 尤溪县林业科技推广中心 | Chinese amanita fuliginea culture medium as well as preparation method and application thereof |
| CN110184244A (en) * | 2019-06-20 | 2019-08-30 | 福清市火麒麟食用菌技术开发有限公司 | A kind of Sparassis crispa produces the preparation method of laccase |
Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JP2005052068A (en) * | 2003-08-04 | 2005-03-03 | Unitika Ltd | Cultivation method for sparasiss crispa |
| KR20070121073A (en) * | 2006-06-21 | 2007-12-27 | 주식회사 비아이지 | The cultivation method of phellinus linteus mycellia having high beta-glucan content |
| CN102696404A (en) * | 2012-07-10 | 2012-10-03 | 云南大学 | Optimized culturing method for promoting growth of Amanitaflavipes mycelia |
-
2013
- 2013-12-19 CN CN201310702241.6A patent/CN103688753B/en not_active Expired - Fee Related
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JP2005052068A (en) * | 2003-08-04 | 2005-03-03 | Unitika Ltd | Cultivation method for sparasiss crispa |
| KR20070121073A (en) * | 2006-06-21 | 2007-12-27 | 주식회사 비아이지 | The cultivation method of phellinus linteus mycellia having high beta-glucan content |
| CN102696404A (en) * | 2012-07-10 | 2012-10-03 | 云南大学 | Optimized culturing method for promoting growth of Amanitaflavipes mycelia |
Non-Patent Citations (3)
| Title |
|---|
| 李宗菊等: "红托鹅膏生态环境研究", 《中国食用菌》 * |
| 王秀建: "平菇菌种的制作技术", 《现代园艺》 * |
| 郭学武等: "三种鹅膏菌培养条件及所产肽类毒素的比较研究", 《菌物学报》 * |
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103980060A (en) * | 2014-06-05 | 2014-08-13 | 尤溪县林业科技推广中心 | Chinese amanita fuliginea culture medium as well as preparation method and application thereof |
| CN103980060B (en) * | 2014-06-05 | 2016-05-25 | 尤溪县林业科技推广中心 | A kind of Chinese Amanita fuliginea culture medium and preparation method thereof and application |
| CN110184244A (en) * | 2019-06-20 | 2019-08-30 | 福清市火麒麟食用菌技术开发有限公司 | A kind of Sparassis crispa produces the preparation method of laccase |
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