CN107189948A - A kind of culture medium and cultural method of promotion flat mushroom liquid mycelia fast-growth - Google Patents
A kind of culture medium and cultural method of promotion flat mushroom liquid mycelia fast-growth Download PDFInfo
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- CN107189948A CN107189948A CN201710423264.1A CN201710423264A CN107189948A CN 107189948 A CN107189948 A CN 107189948A CN 201710423264 A CN201710423264 A CN 201710423264A CN 107189948 A CN107189948 A CN 107189948A
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- C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
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Abstract
The invention belongs to fungus growing technique field, a kind of culture medium and cultural method of promotion flat mushroom liquid mycelia fast-growth are disclosed, cultural method includes：Strain activation and culture, culture matrix manufacturing, cultural hypha, hypha biomass are determined；Cultural hypha punches the hypha of Pleurotus ostreatus grown on flat board with card punch, is then linked into potato dextrose agar (PDA) fluid nutrient medium without agar containing 0.1%~1.0% peanut oil, carries out Shaking culture.The present invention is added after 0.1%~1.0% peanut oil, mycelia can be made dark white, mycelia branch increases, tiny mycelium pellet increase, and dry mycelial weight is not than adding the high by 109.02%~331.96% of peanut oil processing.
Description
Technical field
The invention belongs to fungus growing technique field, more particularly to a kind of training of promotion flat mushroom liquid mycelia fast-growth
Support base and cultural method.
Background technology
Liquid spawn technology is Edible Fungi, the technology especially commonly used in edible fungus industrial production, and it, which has, is permitted
Many advantages：One is that production technology simplifies, and point without parent species, original seed and cultigen, technique is simple, saves time cost；Two are
Strain purity is higher, be bacterial contamination it is small, germination point disperse, bacterium germination is fast；Three be that speed is fast, cost is low for inoculation；Four be production week
Phase is short, and fruiting is neat, is adapted to large-scale production, no seasonal limitation；Five be to be convenient for changing strain, and the replacement cycle is short.
Current factorial praluction liquid spawn is applied to the Pollution In Edible Mushroomss factory metaplasias such as asparagus, pleurotus eryngii, mushroom
In production, and achieve significant economic benefit.Compare other edible fungus varieties, and the application of flat mushroom liquid spawn is less, a side
The production of face and current flat mushroom is that seasonal cultivation production is relevant mostly, another aspect, continuous with people's living standard
Improve, the year-round supply demand for flat mushroom product is continued to increase, and the trend of flat mushroom factorial praluction is irreversible, therefore flat mushroom
The production of liquid spawn is particularly important.Make strain fast-growth within a short period of time and using simple method, concern work
The reduction of factory's operating cost.Strain cultivation influence factor is a lot, such as degree of purity of oxygen content, air etc., culture
The formula of base is also one of mostly important influence factor.
In summary, the problem of prior art is present be：In production, flat mushroom liquid spawn applies less, especially work
Almost do not applied under factory's production model.The formula of existing culture medium is more complicated simultaneously, and formula contained substance species is more,
Manufacture craft is also more complicated.Prior art can not make flat mushroom liquid spawn fast-growth within a short period of time, it is impossible to effectively drop
The reduction of low batch production operating cost.
The content of the invention
The problem of existing for prior art, the invention provides a kind of culture of promotion flat mushroom liquid mycelia fast-growth
Base and cultural method.
The present invention is achieved in that a kind of cultural method of promotion flat mushroom Mycelia fast-growth, the rush
The cultural method for entering flat mushroom Mycelia fast-growth comprises the following steps：
Step one, strain activation and culture:Low temperature or the flat mushroom strain of liquid nitrogen preservation are taken out, potato-grape is seeded in
Activation culture is carried out on sugar-agar (PDA) plating medium；Then beaten with card punch on flat board and take fungus block and be inoculated with fungus block
In the flat board center culture containing PDA culture medium；Beaten again with identical card punch and take fungus block；
Step 2, cultivates matrix manufacturing:Peeled potatoes are cut into small pieces, and are boiled in the pot for being put into the deionized water boiled；So
After filter, glucose is added in filtrate, is allowed to be completely dissolved；Deionized water constant volume is used, is stirred and evenly mixed standby；Then in triangle
The culture medium containing glucose is added in bottle；Peanut oil, ventilated membrane sealing are added in triangular flask；Sterilizing；It is cooled to after room temperature,
It is placed in superclean bench；Beaten with card punch on the mycelia flat board of culture and take multiple strain blocks, then access is free of agar
In Potato-dextrose-agar (PDA) fluid nutrient medium, constant-temperature table, shaken cultivation；
Step 3, hypha biomass is determined:Cultured mycelia is filtered, is then washed with deionized, is then dried to
Constant weight.
Further, step one strain activation and culture, is specifically included：
4 DEG C of low temperature or the flat mushroom strain of liquid nitrogen preservation are taken out, the training of Potato-dextrose-agar (PDA) flat board is seeded in
Support and 25 DEG C of activation culture 8d~10d, repetition of activation 2 times are carried out on base；
Then beaten with diameter 5mm card punch on flat board and take fungus block and be seeded in fungus block flat containing PDA culture medium
Plate center, 25 DEG C of culture 10d；Beaten again with identical card punch and take fungus block.
Further, the step 2 culture matrix manufacturing, is specifically included：
Peeled potatoes 200g, is cut into 0.5cm~1cm fritter, is put into the pot for filling the deionized water that 800mL boils
In, boil 10min~15min；
Then with 4 layers of filtered through gauze, 20g glucose is added in filtrate, is allowed to be completely dissolved；Use deionized water constant volume
To 1000mL, stir and evenly mix standby；
Then 100mL above-mentioned culture medium is added in the triangular flask of 250mL capacity；Quality is added in above-mentioned triangular flask
The peanut oil of volume ratio (w/v) 0.1%~1.0%, to be not added with peanut oil as control, ventilated membrane sealing；121 DEG C of sterilizings
30min；
It is cooled to after room temperature, is placed in superclean bench；Put down with diameter 5mm card punch in culture 8d~12d mycelia
Beaten on plate and take 10 strain blocks, then accessed in fluid nutrient medium, be placed in 25 DEG C of constant-temperature table 150rpm shaken cultivations 5d.
Further, the step 3 hypha biomass is determined, and is specifically included：
By cultured mycelia with 4 layers of filtered through gauze, 3~5 times are then washed with 100mL~200mL deionized water, so
Dried to constant weight afterwards at 50 DEG C.
Another object of the present invention is to provide a kind of culture medium for promoting flat mushroom Mycelia fast-growth by horse
Bell potato 200g, glucose 20g, distilled water 1000mL and in w/v ratios peanut oil 0.1%~1.0% constitute constitute.
Advantages of the present invention and good effect are：
Compared with prior art, the present invention is added after peanut oil, mycelia can be made dark white, mycelia branch increases, can
Promote mycelia fast-growth, dry mycelial weight is not than adding the high by 109.02%~331.96% of peanut oil processing.
Brief description of the drawings
Fig. 1 is the cultural method flow chart of promotion flat mushroom liquid mycelia fast-growth provided in an embodiment of the present invention.
Embodiment
In order to make the purpose , technical scheme and advantage of the present invention be clearer, with reference to embodiments, to the present invention
It is further elaborated.It should be appreciated that the specific embodiments described herein are merely illustrative of the present invention, it is not used to
Limit the present invention.
At present, in production, flat mushroom liquid spawn almost should not using less especially under factorial praluction pattern
With.The formula of existing culture medium is more complicated simultaneously, and formula contained substance species is more, and manufacture craft is also more complicated.It is existing
Technology can not make flat mushroom liquid spawn fast-growth within a short period of time, it is impossible to the effectively reduction of reduction batch production operating cost.
The application principle of the present invention is described in detail below in conjunction with the accompanying drawings.
As shown in figure 1, the cultural method of promotion flat mushroom Mycelia fast-growth provided in an embodiment of the present invention, bag
Include following steps：
S101：Strain activation and culture:Low temperature (4 DEG C) or the flat mushroom strain of liquid nitrogen preservation are taken out, potato-Portugal is seeded in
25 DEG C of activation culture 8d~10d, repetition of activation 2 times are carried out on grape sugar-agar (PDA) plating medium；Then it is (straight with card punch
Footpath 5mm) beaten on flat board take fungus block and by fungus block be seeded in containing PDA culture medium flat board center, 25 DEG C culture 10d；Use again
Identical card punch, which is beaten, takes fungus block；
S102：Cultivate matrix manufacturing:Peeled potatoes 200g, is cut into 0.5cm~1cm or so fritter, is put into and fills 800mL
In the pot for the deionized water boiled, 10min~15min is boiled；Then with 4 layers of filtered through gauze, 20g grape is added in filtrate
Sugar, is allowed to be completely dissolved；1000mL is settled to deionized water, is stirred and evenly mixed standby；Then in the triangular flask of 250mL capacity
Add 100mL above-mentioned culture medium；0.1%~1.0% (w/v) peanut oil is added in above-mentioned triangular flask, to be not added with peanut
Oil is used as control, ventilated membrane sealing；121 DEG C of sterilizing 30min；It is cooled to after room temperature, is placed in superclean bench；Use diameter 5mm
Card punch beat on culture 8d~12d mycelia flat board and take 10 strain blocks, then in access fluid nutrient medium, be placed in 25 DEG C
Constant-temperature table 150rpm shaken cultivations 5d；
S103：Hypha biomass is determined:By cultured mycelia with 4 layers of filtered through gauze, then with 100mL~200mL's
Deionized water is washed 3~5 times, is then dried to constant weight at 50 DEG C.It can draw, peanut oil can remarkably promote the life of hypha of Pleurotus ostreatus
Long, dry mycelial weight is not than adding the high by 109.02%~331.96% of peanut oil processing.
With reference to specific embodiment, the invention will be further described.
Embodiment 1：
The cultural method of promotion flat mushroom Mycelia fast-growth provided in an embodiment of the present invention, including following step
Suddenly：
Strain activation and culture:Low temperature (4 DEG C) or the flat mushroom strain of liquid nitrogen preservation are taken out, be seeded in Potato-dextrose-
25 DEG C of activation culture 8d~10d, repetition of activation 2 times are carried out on agar (PDA) plating medium；Then card punch (diameter is used
5mm) beaten on flat board and take fungus block and fungus block is seeded in the flat board center containing PDA culture medium, 25 DEG C of culture 10d；Phase is used again
Same card punch, which is beaten, takes fungus block；
Cultivate matrix manufacturing:Peeled potatoes 200g, is cut into 0.75cm or so fritter, is put into going of filling that 800mL boils
In the pot of ionized water, 10min~15min is boiled；Then with 4 layers of filtered through gauze, 20g glucose is added in filtrate, is allowed to
It is completely dissolved；1000mL is settled to deionized water, is stirred and evenly mixed standby；Then added in the triangular flask of 250mL capacity
100mL above-mentioned culture medium；0.1% (w/v) peanut oil is added in above-mentioned triangular flask, to be not added with peanut oil as control,
Ventilated membrane is sealed；121 DEG C of sterilizing 30min；It is cooled to after room temperature, is placed in superclean bench；With diameter 5mm card punch in training
Beaten on the mycelia flat board for supporting 8d~12d and take 10 strain blocks, then accessed in fluid nutrient medium, be placed in 25 DEG C of constant-temperature tables
150rpm shaken cultivations 5d；
Hypha biomass is determined:By cultured mycelia with 4 layers of filtered through gauze, then with 100mL~200mL deionization
Water washing 3~5 times, then dries to constant weight at 50 DEG C.It can draw, peanut oil can remarkably promote the growth of hypha of Pleurotus ostreatus, bacterium
Silk dry weight is not than adding the high by 109.02% of peanut oil processing.
Embodiment 2：
The cultural method of promotion flat mushroom Mycelia fast-growth provided in an embodiment of the present invention, including following step
Suddenly：
Strain activation and culture:Low temperature (4 DEG C) or the flat mushroom strain of liquid nitrogen preservation are taken out, be seeded in Potato-dextrose-
25 DEG C of activation culture 8d~10d, repetition of activation 2 times are carried out on agar PDA plate culture medium；Then card punch (diameter 5mm) is used
Beaten on flat board and take fungus block and fungus block is seeded in the flat board center containing PDA culture medium, 25 DEG C of culture 10d；Identical is used again
Card punch, which is beaten, takes fungus block；
Cultivate matrix manufacturing:Peeled potatoes 200g, is cut into 0.5cm or so fritter, be put into going of filling that 800ml boils from
In the pot of sub- water, 10min~15min is boiled；Then with 4 layers of filtered through gauze, 20g glucose is added in filtrate, has been allowed to
Fully dissolved；1000mL is settled to deionized water, is stirred and evenly mixed standby；Then 100mL is added in the triangular flask of 250mL capacity
Above-mentioned culture medium；0.4% (w/v) peanut oil is added in above-mentioned triangular flask, to be not added with peanut oil as control, ventilated membrane
Sealing；121 DEG C of sterilizing 30min；It is cooled to after room temperature, is placed in superclean bench；With diameter 5mm card punch culture 8d~
Beaten on 12d mycelia flat board and take 10 strain blocks, then accessed in fluid nutrient medium, be placed in 25 DEG C of constant-temperature table 150rpm vibrations
Cultivate 5d；
Hypha biomass is determined:By cultured mycelia with 4 layers of filtered through gauze, then with 100mL~200mL deionization
Water washing 3~5 times, then dries to constant weight at 50 DEG C.It can draw, peanut oil can remarkably promote the growth of hypha of Pleurotus ostreatus, bacterium
Silk dry weight is not than adding the high by 161.54% of peanut oil processing.
Embodiment 3：
The cultural method of promotion flat mushroom Mycelia fast-growth provided in an embodiment of the present invention, including following step
Suddenly：
Strain activation and culture:Low temperature (4 DEG C) or the flat mushroom strain of liquid nitrogen preservation are taken out, be seeded in Potato-dextrose-
25 DEG C of activation culture 8d~10d, repetition of activation 2 times are carried out on agar PDA plate culture medium；Then card punch (diameter 5mm) is used
Beaten on flat board and take fungus block and fungus block is seeded in the flat board center containing PDA culture medium, 25 DEG C of culture 10d；Identical is used again
Card punch, which is beaten, takes fungus block；
Cultivate matrix manufacturing:Peeled potatoes 200g, is cut into 1cm or so fritter, is put into and fills the deionization that 800mL boils
In the pot of water, 10min~15min is boiled；Then with 4 layers of filtered through gauze, 20g glucose is added in filtrate, is allowed to complete
Dissolving；1000mL is settled to deionized water, is stirred and evenly mixed standby；Then add 100mL's in the triangular flask of 250mL capacity
Above-mentioned culture medium；1.0% (w/v) peanut oil is added in above-mentioned triangular flask, to be not added with peanut oil as control, ventilated membrane envelope
Mouthful；121 DEG C of sterilizing 30min；It is cooled to after room temperature, is placed in superclean bench；With diameter 5mm card punch culture 8d~
Beaten on 12d mycelia flat board and take 10 strain blocks, then accessed in fluid nutrient medium, be placed in 25 DEG C of constant-temperature table 150rpm vibrations
Cultivate 5d；
Hypha biomass is determined:By cultured mycelia with 4 layers of filtered through gauze, then with 100mL~200mL deionization
Water washing 3~5 times, then dries to constant weight at 50 DEG C.It can draw, peanut oil can remarkably promote the growth of hypha of Pleurotus ostreatus, bacterium
Silk dry weight is not than adding the high by 331.96% of peanut oil processing.
The foregoing is merely illustrative of the preferred embodiments of the present invention, is not intended to limit the invention, all essences in the present invention
Any modifications, equivalent substitutions and improvements made within refreshing and principle etc., should be included in the scope of the protection.
Claims (5)
1. a kind of cultural method of promotion flat mushroom Mycelia fast-growth, it is characterised in that the promotion flat mushroom liquid
The cultural method of culture mycelia fast-growth comprises the following steps：
Step one, strain activation and culture:Low temperature or the flat mushroom strain of liquid nitrogen preservation are taken out, Potato-dextrose-fine jade is seeded in
Activation culture is carried out on fat plating medium；Then beaten with card punch on flat board and take fungus block and fungus block is seeded in containing PDA
The flat board center culture of culture medium；Beaten again with identical card punch and take fungus block；
Step 2, cultivates matrix manufacturing:Peeled potatoes are cut into small pieces, and are boiled in the pot for being put into the deionized water boiled；Then cross
Filter, glucose is added in filtrate, is allowed to be completely dissolved；Deionized water constant volume is used, is stirred and evenly mixed standby；Then in triangular flask
Add the culture medium containing glucose；Peanut oil, ventilated membrane sealing are added in triangular flask；Sterilizing；It is cooled to after room temperature, is placed in
In superclean bench；Beaten with card punch on the mycelia flat board of culture and take multiple strain blocks, then Ma Ling of the access without agar
Potato-glucose-agar) in fluid nutrient medium, constant-temperature table, shaken cultivation；
Step 3, hypha biomass is determined:Cultured mycelia is filtered, is then washed with deionized, is then dried to constant weight.
2. promote the cultural method of flat mushroom Mycelia fast-growth as claimed in claim 1, it is characterised in that described
Step one strain activation and culture, is specifically included：
4 DEG C of low temperature or the flat mushroom strain of liquid nitrogen preservation are taken out, Potato-dextrose-Agar Plating are seeded in enterprising
25 DEG C of activation culture 8d~10d of row, repetition of activation 2 times；
Then beaten with diameter 5mm card punch on flat board and take fungus block and fungus block is seeded in the flat board containing PDA culture medium
Centre, 25 DEG C of culture 10d；Beaten again with identical card punch and take fungus block.
3. promote the cultural method of flat mushroom Mycelia fast-growth as claimed in claim 1, it is characterised in that described
Step 2 culture matrix manufacturing, is specifically included：
Peeled potatoes 200g, is cut into 0.5cm~1cm fritter, is put into the pot for filling the deionized water that 800mL boils, boils
Boil 10min~15min；
Then with 4 layers of filtered through gauze, 20g glucose is added in filtrate, is allowed to be completely dissolved；It is settled to deionized water
1000mL, is stirred and evenly mixed standby；
Then 100mL above-mentioned culture medium is added in the triangular flask of 250mL capacity；Quality volume is added in above-mentioned triangular flask
Than the peanut oil that w/v is 0.1%~1.0%, to be not added with peanut oil as control, ventilated membrane sealing；121 DEG C of sterilizing 30min；
It is cooled to after room temperature, is placed in superclean bench；With diameter 5mm card punch on culture 8d~12d mycelia flat board
Beat and take 10 strain blocks, then access in fluid nutrient medium, be placed in 25 DEG C of constant-temperature table 150rpm shaken cultivations 5d.
4. promote the cultural method of flat mushroom Mycelia fast-growth as claimed in claim 1, it is characterised in that described
Step 3 hypha biomass is determined, and is specifically included：
By cultured mycelia with 4 layers of filtered through gauze, then 3~5 times, Ran Hou are washed with 100mL~200mL deionized water
50 DEG C dry to constant weight.
5. a kind of culture medium of the cultural method culture of promotion flat mushroom Mycelia fast-growth as claimed in claim 1,
Characterized in that, the culture medium for promoting flat mushroom Mycelia fast-growth is by potato 200g, glucose 20g, steaming
Distilled water 1000mL and the composition of peanut oil 0.1%~1.0% in w/v ratios.
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| CN201710423264.1A CN107189948A (en) | 2017-06-07 | 2017-06-07 | A kind of culture medium and cultural method of promotion flat mushroom liquid mycelia fast-growth |
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| CN201710423264.1A CN107189948A (en) | 2017-06-07 | 2017-06-07 | A kind of culture medium and cultural method of promotion flat mushroom liquid mycelia fast-growth |
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Cited By (5)
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| CN109769590A (en) * | 2019-02-25 | 2019-05-21 | 黔西县盛腾农业科技开发有限责任公司 | A method of oyster mushroom is planted using mushroom waste material |
| CN111394252A (en) * | 2020-03-27 | 2020-07-10 | 江苏华绿生物科技股份有限公司 | Preservation strain activation transfer method |
| CN114503873A (en) * | 2020-11-16 | 2022-05-17 | 上海国森生物科技有限公司 | Agaricus bisporus liquid strain reduction strain and cultivation method thereof |
| CN114868599A (en) * | 2022-06-09 | 2022-08-09 | 河南省农业科学院植物营养与资源环境研究所 | Method for accelerating induction of formation of pleurotus ostreatus primordium by using maltose |
| WO2023116563A1 (en) * | 2021-12-21 | 2023-06-29 | 深圳先进技术研究院 | Leather-like biological composite material, preparation method therefor and use thereof |
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| Publication number | Priority date | Publication date | Assignee | Title |
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| CN114503873A (en) * | 2020-11-16 | 2022-05-17 | 上海国森生物科技有限公司 | Agaricus bisporus liquid strain reduction strain and cultivation method thereof |
| WO2023116563A1 (en) * | 2021-12-21 | 2023-06-29 | 深圳先进技术研究院 | Leather-like biological composite material, preparation method therefor and use thereof |
| CN114868599A (en) * | 2022-06-09 | 2022-08-09 | 河南省农业科学院植物营养与资源环境研究所 | Method for accelerating induction of formation of pleurotus ostreatus primordium by using maltose |
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