CN101849476A - Method for rapidly producing agaric strain by sclerotizing - Google Patents
Method for rapidly producing agaric strain by sclerotizing Download PDFInfo
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- CN101849476A CN101849476A CN200910010974A CN200910010974A CN101849476A CN 101849476 A CN101849476 A CN 101849476A CN 200910010974 A CN200910010974 A CN 200910010974A CN 200910010974 A CN200910010974 A CN 200910010974A CN 101849476 A CN101849476 A CN 101849476A
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- umbellate pore
- pore furgus
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- halimasch
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Abstract
The invention relates to Chinese medicinal agaric, in particular to a method for rapidly producing an agaric strain by sclerotizing. The method comprises the following steps of: 1) preparing a first-stage culture medium from 10 to 50 grams of corn flour, 10 to 20 grams of cane sugar or glucose, 1 to 3 grams of beef extract, 1,000 milliliters of water and 15 to 22 grams of agar, sterilizing the first-stage culture medium, inoculating the agaric strain and performing first-stage culture of the agaric to obtain a first-stage strain; and 2) mixing 20 to 60 weight percent of corn grit and 80 to 40 weight percent of broadleaf tree sawdust and/or a small wood section of which the diameter is between 0.1 and 2 centimeters and the length is between 3 and 5 centimeters to prepare a second-stage culture medium, adding water into the second-stage culture medium, uniformly stirring the mixture until the water content of the culture medium reaches 55 to 60 percent, sterilizing the mixture, simultaneously inoculating the first-stage strain and halimasch of the agaric strain and cultivating the strain at the temperature of between 20 and 25 DEG C for 30 to 45 days to obtain a second-stage strain which can be propagated in a great quantity. The method has the advantages of low production cost and rapid sclerotium formation.
Description
Technical field
The present invention relates to the traditional Chinese medicine umbellate pore furgus, the quick production method of specifically a kind of umbellate pore furgus bacterial classification sclerotiumization.
Background technology
Umbellate pore furgus is the traditional rare traditional Chinese medicine of China, have effects such as diuresis excreting dampness, treating stranguria detumescence, at present the umbellate pore furgus of pharmaceutically using is many by excavating wild product or adopting wild sclerotium to make kind of half an artificial cultivation 3-4 gather (document 1: Guo Shunxing, Xu Jintang, Xiao Peigen.Umbellate pore furgus biological study progress.CHINA JOURNAL OF CHINESE MATERIA MEDICA, 1996, (9): 515-517; Document 2: Xu Guangbo, Fu Weijie, Zhao Xukui.China's umbellate pore furgus progress of research.Fungus research, 2003,1 (1): 58-61; Document 3: Cheng Xianhao.Umbellate pore furgus and halimasch artificial culture and influence the research that several active ingredients produce factors.China's doctorate paper, 2006-09-27:1-181).Because the expansion and the outlet of medical market, wild product resource falls sharply, and is difficult to meet the need of market.Adopt wild sclerotium to make the planting type of planting, need a large amount of wild sclerotium (grey Siberian cocklebur is the 2nd year product of umbellate pore furgus), cause artificial cultivation provenance difficulty, cause the artificial cultivation cost to rise, become the main bottleneck (document 4: Xing Xiaoke that restriction umbellate pore furgus artificial cultivation technique large tracts of land is implemented.Umbellate pore furgus with mix living bacterium Study of Interaction.China's doctorate paper, 2006-10-13:1-154).Therefore, high efficiency, low cost breeding problem extremely urgent (document 5: Guo Shunxing, Wang Qiuying, Zhang Jihui, full in summer swallow during solution umbellate pore furgus bacterial classification is produced.The umbellate pore furgus mycelia forms the research of sclerotium cultivation method.Chinese Pharmaceutical Journal, 2001,36 (10): 559-560; Document 6: Xu Guangbo, Fu Weijie, Li Ruliang etc.The research of Changbaishan area umbellate pore furgus half artificial cultivation technique system.Yanbian University's agronomy journal, 2004,24 (4): 241-244).
Summary of the invention
The object of the present invention is to provide the quick production method of a kind of umbellate pore furgus bacterial classification sclerotiumization.
For achieving the above object, the technical solution used in the present invention is:
The quick production method of a kind of umbellate pore furgus bacterial classification sclerotiumization,
1) the one-level culture medium prescription consists of: 10-50g corn flour, 10-20g sucrose or glucose, beef extract 1-3g, water 1000ml, agar 15-22g; Sterilization back inoculation umbellate pore furgus bacterial classification carries out the one-level of umbellate pore furgus bacterium and cultivates, and gets first class inoculum;
2) by weight, with the 20-60% corn cook and stir, the little juggle of 80-40% hardwood sawdust and/or diameter 0.1-2cm, long 3-5cm is mixed and made into secondary medium, adds water and stirs, and makes the weight water content of medium reach 55-60%; Inoculate the one-level kind and the halimasch of umbellate pore furgus bacterial classification after the sterilization simultaneously, cultivated 30-45 days under 20-25 ℃ of condition, promptly obtaining can prolific second class inoculum.
The secondary medium of water content 55-60% is packed in the polypropylene plastics pocket into sterilization back inoculation step 2) second class inoculum that obtains, under 20-25 ℃ of condition, cultivated 30-45 days, promptly obtaining can prolific three-class strain.
The condition of culture of umbellate pore furgus bacterium is in the step 1), and be 16-25 with the umbellate pore furgus bacterial classification with the volume ratio with medium: 100 ratio is mixed, and cultivates 14-20 days for 20-27 ℃, forms a large amount of sclerotium on the test tube slant, gets first class inoculum.
Described step 2) umbellate pore furgus bacterial classification and the halimasch inoculum concentration in secondary medium is, umbellate pore furgus bacterial classification and halimasch are 0.5-5 with the volume ratio of medium respectively: 100.
The halimasch of being adopted step 2) is the halimasch of adopting after the one-level medium culture is bred, and its condition of culture is that be 16-25 with the halimasch bacterial classification with the volume ratio with medium: 100 ratio is mixed, and cultivates 14-20 days for 20-27 ℃, gets halimasch.
When described secondary medium was prepared, corn is cooked and stir needed water in advance to soak 12-16 hour, prewets and mixes with hardwood sawdust and/or little juggle; Described broad-leaved tree is one or more in toothed oak tree, Chinese chestnut, maple, the elm.
The present invention has following advantage:
1. production cost is low.Adopt wild sclerotia half artificial culture method cultivation, every mu of sclerotia cost is 15000 yuan (calculating with the wild sclerotium of 300kg, cultivation 600 caves by every mu).Because sclerotia price height is difficult to again guarantee to produce a large amount of needs and supply in time simultaneously, has greatly restricted the umbellate pore furgus commerial growing.Materials such as the umbellate pore furgus bacterial classification that utilizes the inventive method to produce carries out artificial cultivation, and every mu needs umbellate pore furgus bacterial classification 300kg, 600 yuan of costs, cultivation 600 caves, other bacterium rods and cost are identical with in the past cultivation method.
Adopt the umbellate pore furgus bacterial classification that the present invention produced to carry out artificial cultivation, can save 14400 yuan than conventional method for every mu, the cultivation cost is 4% of a conventional method, has tangible economic benefit, can satisfy the needs of large-scale production efficiently, in time.
2. sclerotium forms soon.Cultivated the umbellate pore furgus bacterial classification to adopt the PDA medium be the one-level medium more in the past, and the umbellate pore furgus mycelial growth is slow, it is few to form sclerotium, and sclerotium forms to be needed more than 30 days.To the research of umbellate pore furgus C, N source, the umbellate pore furgus bacterial classification one-level culture medium prescription of being developed can form a large amount of sclerotium at 14 days through the applicant, than PDA medium fast 16 days, had solved enlarged culture prescription and three grades of umbellate pore furgus culture propagation technology of second class inoculum simultaneously.Inoculation umbellate pore furgus bacterium and halimasch when the expansion of secondary kind is numerous, can form in 35-40 days can be for the umbellate pore furgus bacterial classification of using in producing, owing to use corn flour and lower cost materials such as corn is cooked and stir, sucrose wood chip and little juggle are produced the umbellate pore furgus bacterial classification that can form sclerotium in a large number, the wild sclerotium that uses in the half artificial cultivation process of substituting has greatly reduced production cost.
Embodiment
Embodiment 1
1, with 10g corn flour, 20g sucrose, beef extract 3g, water 1000ml, agar 22g, preparation one-level medium; Under 1.5 atmospheric pressure, 121 ℃ of conditions, sterilized 20 minutes;
1) inoculation umbellate pore furgus bacterial classification under gnotobasis, the volume ratio of umbellate pore furgus bacterial classification and medium is 25: 100, cultivates for 20 ℃ and makes first class inoculum in 20 days, forms a large amount of sclerotium on the test tube slant;
2) with after the sterilization of one-level culture medium prescription, 25 ℃ of upright test tube coolings, the switching halimasch, the volume ratio of umbellate pore furgus bacterial classification and medium is 25: 100, condition of culture gets the halimasch bacterial classification with the umbellate pore furgus bacterial classification;
2, with 20% corn cook and stir, the part by weight of 80% toothed oak trees bits is mixed with the secondary medium of water content 60%, corn is cooked and stir before the use in advance through 12-16 hour soaked prewetting; Then secondary medium is packed into Cans and seal with polypropylene film, under 121 ℃ of conditions, autoclaving 90 minutes; To be cooled to below 25 ℃, umbellate pore furgus first class inoculum and halimasch bacterial classification that while inoculation step " 1 " obtains, the umbellate pore furgus bacterial classification of inoculation and halimasch are 2: 100 with the volume ratio of medium respectively, cultivate 45 days under 20 ℃ of conditions, and promptly obtaining can prolific second class inoculum;
3, with the polypropylene spear pocket secondary medium of packing into, after the method for " 2 " sterilization, the inoculation second class inoculum was also cultivated 45 days set by step, and the second class inoculum of inoculation and the volume ratio of medium are 2: 100, the method of the same step of condition of culture " 2 " promptly obtains three-class strain.
Embodiment 2
1, with 50g corn flour, 10g glucose, beef extract 1g, water 1000ml, agar 15g, preparation one-level medium; Under 1.5 atmospheric pressure, 121 ℃ of conditions, sterilized 20 minutes;
1) inoculation umbellate pore furgus bacterial classification under gnotobasis, the volume ratio of umbellate pore furgus bacterial classification and medium is 20: 100, cultivates for 27 ℃ and makes first class inoculum in 20 days, forms a large amount of sclerotium on the test tube slant;
2) with after the sterilization of one-level culture medium prescription, 25 ℃ of upright test tube coolings, the switching halimasch, the volume ratio of umbellate pore furgus bacterial classification and medium is 20: 100, condition of culture is with the umbellate pore furgus bacterial classification;
2, with part by weight 60% corn cook and stir, the little juggle of 40% broad-leaved tree maple and elm diameter 0.1-2cm, long 3-5cm is mixed with the secondary medium of water content 55%, corn is cooked and stir before the use in advance through 12-16 hour soaked prewetting; Then secondary medium is packed into Cans and seal with polypropylene film, under 121 ℃ of conditions, autoclaving 90 minutes; To be cooled to below 25 ℃, umbellate pore furgus first class inoculum and halimasch bacterial classification that while inoculation step " 1 " obtains, the umbellate pore furgus first class inoculum of inoculation and halimasch are 4: 100 with the volume ratio of medium respectively, cultivate 30 days under 25 ℃ of conditions, and promptly obtaining can prolific second class inoculum;
3, with the polypropylene spear pocket secondary medium of packing into, after the method for " 2 " sterilization, the inoculation second class inoculum was also cultivated 35 days set by step, and the second class inoculum of inoculation and the volume ratio of medium are 5: 100, the method of the same step of condition of culture " 2 " promptly obtains three-class strain.
Embodiment 3
1, with 30g corn flour, 15g sucrose or glucose, beef extract 2g, water 1000ml, agar 18g, preparation one-level medium; Under 1.5 atmospheric pressure, 121 ℃ of conditions, sterilized 20 minutes;
1) inoculation umbellate pore furgus bacterial classification under gnotobasis, the volume ratio of umbellate pore furgus bacterial classification and medium is 16: 100, cultivates for 25 ℃ and makes first class inoculum in 18 days, forms a large amount of sclerotium on the test tube slant;
2) with after the sterilization of one-level culture medium prescription, 25 ℃ of upright test tube coolings, the switching halimasch, the volume ratio of umbellate pore furgus bacterial classification and medium is 16: 100, condition of culture is with the umbellate pore furgus bacterial classification;
2, with 30% corn cook and stir, the part by weight of 70% Chinese chestnut wood chip is mixed with the secondary medium of water content 58%, corn is cooked and stir needs 12-16 hour soaked prewetting in advance; Then secondary medium is packed into Cans and seal with polypropylene film, under 121 ℃ of conditions, autoclaving 90 minutes; To be cooled to below 25 ℃, umbellate pore furgus first class inoculum and halimasch bacterial classification that while inoculation step " 1 " obtains, the umbellate pore furgus first class inoculum of inoculation and halimasch are 1: 100 with the volume ratio of medium respectively, cultivate 40 days under 23 ℃ of conditions, and promptly obtaining can prolific second class inoculum;
3, with the polypropylene spear pocket secondary medium of packing into, after the method for " 2 " sterilization, the inoculation second class inoculum was also cultivated 40 days set by step, and the second class inoculum of inoculation and the volume ratio of medium are 1: 100, the method of the same step of condition of culture " 2 " promptly obtains three-class strain.
Second class inoculum that the above-mentioned example of the present invention is obtained or three-class strain promptly can be used for the cultivation of umbellate pore furgus yielding ability.
Claims (7)
1. quick production method of umbellate pore furgus bacterial classification sclerotiumization is characterized in that:
1) the one-level culture medium prescription consists of: 10-50g corn flour, 10-20g sucrose or glucose, beef extract 1-3g, water 1000ml, agar 15-22g; Sterilization back inoculation umbellate pore furgus bacterial classification carries out the one-level of umbellate pore furgus bacterium and cultivates, and gets first class inoculum;
2) by weight, with the 20-60% corn cook and stir, the little juggle of 80-40% hardwood sawdust and/or diameter 0.1-2cm, long 3-5cm is mixed and made into secondary medium, adds water and stirs, and makes the weight water content of medium reach 55-60%; Inoculate the one-level kind and the halimasch of umbellate pore furgus bacterial classification after the sterilization simultaneously, cultivated 30-45 days under 20-25 ℃ of condition, promptly obtaining can prolific second class inoculum.
2. according to the described production method of claim 1, it is characterized in that: the secondary medium of water content 55-60% is packed in the polypropylene plastics pocket, sterilization back inoculation step 2) second class inoculum that obtains was cultivated 30-45 days under 20-25 ℃ of condition, and promptly obtaining can prolific three-class strain.
3. according to the described production method of claim 1, it is characterized in that: the condition of culture of umbellate pore furgus bacterium is in the step 1), and be 16-25 with the umbellate pore furgus bacterial classification with the volume ratio with medium: 100 ratio is mixed, and cultivates 14-20 days for 20-27 ℃, on the test tube slant, form a large amount of sclerotium, get first class inoculum.
4. according to the described production method of claim 1, it is characterized in that: described step 2) umbellate pore furgus bacterial classification and the halimasch inoculum concentration in secondary medium is, umbellate pore furgus bacterial classification and halimasch are 0.5-5 with the volume ratio of medium respectively: 100.
5. according to the described production method of claim 1, it is characterized in that: step 2) in the halimasch of being adopted be the halimasch of adopting after the one-level medium culture propagation, its condition of culture is, is 16-25 with the halimasch bacterial classification with the volume ratio with medium: 100 ratio is mixed, cultivated 14-20 days for 20-27 ℃, get halimasch.
6. according to the described production method of claim 1, it is characterized in that: when described secondary medium was prepared, corn is cooked and stir needed water in advance to soak 12-16 hour, prewets and mixes with hardwood sawdust and/or little juggle.
7. according to the described production method of claim 1, it is characterized in that: described broad-leaved tree is one or more in toothed oak tree, Chinese chestnut, maple, the elm.
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| CN2009100109747A CN101849476B (en) | 2009-04-01 | 2009-04-01 | Method for rapidly producing agaric strain by sclerotizing |
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| CN2009100109747A CN101849476B (en) | 2009-04-01 | 2009-04-01 | Method for rapidly producing agaric strain by sclerotizing |
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Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102812854A (en) * | 2012-09-10 | 2012-12-12 | 薛向阳 | Manual wild-imitation cultivation technology for pure strains of grifola |
| CN105900685A (en) * | 2016-05-05 | 2016-08-31 | 甄永福 | Grifola umbellate and pteridium aquilinum mixed planting method |
| CN107759306A (en) * | 2016-08-22 | 2018-03-06 | 鲁克千 | The method of vacuum collecting umbellate pore furgus strain |
| CN112840949A (en) * | 2021-01-25 | 2021-05-28 | 中国医学科学院药用植物研究所 | Artificial cultivation method of polyporus umbellatus |
Family Cites Families (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN100457891C (en) * | 2005-07-22 | 2009-02-04 | 中国医学科学院药用植物研究所 | New method for cultivating umbellate pore fungus mycelium to form umbellate pore fungus sclerotium |
-
2009
- 2009-04-01 CN CN2009100109747A patent/CN101849476B/en not_active Expired - Fee Related
Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102812854A (en) * | 2012-09-10 | 2012-12-12 | 薛向阳 | Manual wild-imitation cultivation technology for pure strains of grifola |
| CN105900685A (en) * | 2016-05-05 | 2016-08-31 | 甄永福 | Grifola umbellate and pteridium aquilinum mixed planting method |
| CN107759306A (en) * | 2016-08-22 | 2018-03-06 | 鲁克千 | The method of vacuum collecting umbellate pore furgus strain |
| CN112840949A (en) * | 2021-01-25 | 2021-05-28 | 中国医学科学院药用植物研究所 | Artificial cultivation method of polyporus umbellatus |
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