CN101731097A - Grifola frondosus liquid strain and method for cultivating grifola frondosus by using liquid strain - Google Patents
Grifola frondosus liquid strain and method for cultivating grifola frondosus by using liquid strain Download PDFInfo
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- CN101731097A CN101731097A CN200910201986A CN200910201986A CN101731097A CN 101731097 A CN101731097 A CN 101731097A CN 200910201986 A CN200910201986 A CN 200910201986A CN 200910201986 A CN200910201986 A CN 200910201986A CN 101731097 A CN101731097 A CN 101731097A
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Abstract
The invention discloses a grifola frondosus liquid strain and a method for cultivating grifola frondosus by using the liquid strain. The liquid strain has good dispersion and strong fluidity, and can quicken the growth speed of the strain and shorten the fermentation time; and bag hypha prepared from the liquid strain grows healthily, the growth of each part is uniform, the fungus age is regular, and the grown mushroom is regular and facilitates factory management.
Description
Technical field:
The present invention relates to field of edible fungus culture, relate in particular to a kind of grifola frondosus liquid spawn and with the method for this liquid spawn fast breeding technique cultivation grifola frondosus.
Background technology:
With the liquid strain cultivation edible mushroom is the new technology that occurred in recent years, and " liquid spawn " is comparatively speaking with " solid spawn ", refers to the edible fungus species that obtains by the deep layer culture technique with liquid nutrient medium.Compare with traditional solid spawn, liquid spawn can breed out the good quality strain of a large amount of high-purities, high vigor cheaply in the short time, and can easily it directly be seeded in the bacterium bag.Liquid spawn has been used for mushroom kinds such as cultivating flammulina velutipes, hypsizygus marmoreus, flat mushroom at present, and has formed industrialization in the factory culture Asparagus, yet there are no report and the liquid spawn fast breeding technique is applied to the grifola frondosus factory culture.
Summary of the invention:
Technical problem to be solved by this invention is to provide a kind of grifola frondosus liquid spawn and cultivates the method for grifola frondosus with it.
The present invention at first provides a kind of liquid spawn of grifola frondosus, and it prepares by the following method:
The female kind made: grifola frondosus mycelia piece is inserted the PDA medium, and 25 ℃ of dark culturing to mycelia are covered with the inclined-plane;
The level liquid bacterial classification is made: the culture fluid triangular flask of packing into, be cooled to after the sterilization below 25 ℃, and insert 3-5cm
2Mother's kind of smashing leaves standstill under 24-25 ℃ and cultivates 48h, places the shaking table shaken cultivation again, and frequency of oscillation is 100-150r/min, and the shaking table room temperature is cultivated 7-10d at 24-25 ℃;
Second-class liquid isolate is made: culture fluid is packed in the fermentation tank, and culture fluid is sterilized with the aeration equipment of fermentation tank, is cooled to after the sterilization below 25 ℃, inserts the level liquid bacterial classification, cultivates 12-15d, promptly obtains the grifola frondosus liquid spawn.
The present invention also provides the method with above-mentioned liquid strain cultivation grifola frondosus, and this method comprises the steps:
The grifola frondosus cultivating bacteria bag is made: the pack of medium wet feed, and every packed wet feed 700g-900g, high 12cm-14cm, sterilization, cooling, every bag of liquid spawn that meets 20-25ml places temperature 22-25 ℃, and relative moisture is 60%-70%, CO
2Concentration is to cultivate under the 1000-2000ppm condition, cultivates 25-30d;
The fruiting cultivation management: 10d afterripening after the mycelia purseful, treat that mycelia walks the back fully at lateral opening; Place temperature 22-24 ℃, relative moisture 95%-99%, the darkroom stimulates flower bud; The existing spherical original hase of 6-8d aperture position; Be provided with temperature 18-20 ℃, relative moisture 90%-95%, CO
2Concentration 500-800ppm, spherical original hase changes into the brain shape, treats that fruit body length is fan-shaped to cap, and can gather on the first appearance in the bacterium hole.
Grifola frondosus liquid spawn provided by the present invention and with liquid spawn fast breeding technique cultivation grifola frondosus can significantly shorten grifola frondosus bacterial classification fabrication cycle.The liquid spawn good dispersion, mobile strong, can penetrate into each position of solid culture medium with culture fluid, form many bacterium centers, accelerate the growth rate of bacterial classification, thereby shortened a bacterium time greatly.And improved the shortcoming of using top and the bottom mycelia cell age difference in the solid spawn bacterium bag.The bacterium bag hyphal development stalwartness that adopts liquid spawn to make, each several part growth uniformity, cell age is neat, be convenient to the later stage can urge the same period flower bud to handle, and fruiting is neat, is convenient to factory management.Liquid spawn fast breeding technique provided by the present invention is applicable to grifola frondosus factory culture pattern, and provides reference for other cultivars such as mushroom kinds such as hypsizygus marmoreus, Xingbao mushroom.
Embodiment
Grifola frondosus liquid mycelia provided by the present invention prepares by the following method:
The female kind made: grifola frondosus mycelia piece is inserted the PDA medium, and mycelia can be covered with the inclined-plane behind 25 ℃ of dark culturing 10d;
The level liquid bacterial classification is made: adopt the shake-flask culture mode.Culture fluid prescription: potato 5%, glucose 2%, corn flour 2-5%, wheat bran leaching liquor 1-3%, agar 0.8-1%, pH nature.After culture fluid prepared, in the triangular flask of the 500mL capacity of packing into, every bottled amount was 100mL, and adds 10 beades, added to wrap up brown paper again behind the tampon and seal, and when being cooled to below 25 ℃ after the sterilization, inserted 3-5cm
2With mother's kind that refiner is smashed, under 25 ℃, leave standstill and cultivate 48h, place the shaking table shaken cultivation again, frequency of oscillation is 100-150r/min.The shaking table room temperature is controlled at 24 ℃～25 ℃, and it is as clear as crystal to culture fluid to cultivate 7-10d, and a large amount of Grifolas frondosa germ pompons are suspending in the liquid.
Second-class liquid isolate (being liquid spawn) is made: adopt the grifola frondosus liquid fermentation tank.After culture fluid prepares, pack in the fermentation tank, every bottled amount is 3-5L, the aeration equipment of culture fluid and fermentation tank is together sterilized, and when being cooled to below 25 ℃ after the sterilization, inserts the level liquid bacterial classification of 200-400ml, open inlet duct and constantly replenish the fresh air that has filtered, it is as clear as crystal to culture fluid to cultivate 12-15d, and a large amount of Grifolas frondosa germ pompons that suspending in the liquid are the grifola frondosus liquid spawn.
Concrete grammar with above-mentioned liquid strain cultivation grifola frondosus is as follows:
The grifola frondosus cultivating bacteria bag is made: the culture medium raw material proportioning is: wood chip 30%, cotton seed hulls 30%, wheat bran 20%, corn flour 5%, brown sugar 1%, gypsum 1%, soil 13%.Water content 62%-65% is with automatic packer pack, every packed wet feed 700g-900g, high 12cm-14cm.The every bag of liquid spawn that meets 20-25ml in sterilization back is placed on temperature 22-25 ℃, and relative moisture is 60%-70%, CO
2Concentration is to cultivate under the condition of 1000-2000ppm, and the 25-30d mycelia can be covered with the bacterium bag.
Fruiting cultivation management: need the 10d afterripening after the mycelia purseful, treat that mycelia walks the back fully at the side scarfing.Place temperature 22-24 ℃, relative moisture 95%-99%, the darkroom stimulates flower bud; The existing spherical original hase in 7d after-opening position; Be provided with temperature 18-20 ℃, relative moisture 90%-95%, CO
2Concentration 500-800ppm,, indoor fluorescent lamp is opened; Spherical original hase is divided into the brain shape gradually, treats that fruit body length is fan-shaped to cap, can gather on the first appearance in the bacterium hole.
Embodiment one:
The female kind made: grifola frondosus bacterial classification piece (preservation of DSMZ of Edible Fungus Inst., Shanghai Academy of Agriculture, numbering: GF1) be inoculated in the PDA medium, 25 ℃ of dark culturing 10d;
The level liquid bacterial classification is made (shake-flask culture mode): the culture fluid prescription: potato 5%, glucose 2%, corn flour 3%, wheat bran leaching liquor 1%, agar 0.8%, pH nature; After culture fluid prepares, pack in the triangular flask of 500mL capacity, every bottled amount is 100mL, and add 10 beades, add and wrap up brown paper again behind the tampon and seal, when being cooled to below 25 ℃ after the sterilization, mother's kind that access is smashed with refiner, leave standstill under 25 ℃ and cultivate 48h, place the shaking table shaken cultivation again, frequency of oscillation is 100r/min.It is as clear as crystal to cultivate the visible culture fluid of 10d, and a large amount of Grifolas frondosa germ pompons are suspending in the liquid;
Second-class liquid isolate (being liquid spawn) is made (liquid fermentation tank cultivation): the culture fluid prescription is the same, after culture fluid prepares, pack in the fermentation tank, every bottled amount is 3L, and the aeration equipment of culture fluid and fermentation tank is together sterilized, when being cooled to below 25 ℃ after the sterilization, insert the level liquid bacterial classification of 200ml, open inlet duct and constantly replenish the fresh air that has filtered, it is as clear as crystal to cultivate the visible culture fluid of 14d, and a large amount of Grifolas frondosa germ pompons are suspending in the liquid; Obtain liquid spawn.
The grifola frondosus cultivating bacteria bag is made: the culture medium raw material proportioning is: wood chip 30%, cotton seed hulls 30%, wheat bran 20%, corn flour 5%, brown sugar 1%, gypsum 1%, soil 13%.Add water and stir, making its water content is 65%, with automatic packer pack, every packed wet feed 900g, high 14cm.The every bag of liquid spawn that meets 20ml in sterilization back is placed on 22 ℃ of temperature, and relative moisture is 60%%, CO
2Concentration is that 1000ppm cultivates, and the 28d mycelia is promptly covered with the bacterium bag;
Fruiting cultivation management: need the 10d afterripening after the mycelia purseful, treat that mycelia walks the back fully at the side scarfing.Place 22 ℃ of temperature, relative moisture 95%, the darkroom stimulates flower bud.The existing spherical original hase in 7d after-opening position.19 ℃ of temperature, relative moisture 95%, CO are set this moment
2Concentration 500ppm, indoor fluorescent lamp is opened.Spherical original hase is divided into the brain shape gradually, treats that fruit body length is fan-shaped to cap, can gather on the first appearance in the bacterium hole.
Embodiment two:
The female kind made with embodiment one; Used bacterial classification is GF2 (preservation of DSMZ of Edible Fungus Inst., Shanghai Academy of Agriculture, a numbering: GF2)
Level liquid bacterial classification system (shake-flask culture mode): culture fluid prescription: potato 5%, glucose 2%, corn flour 2%, wheat bran leaching liquor 3%, agar 1%, pH nature.Bottling, sterilization, inoculation are with embodiment one, and inoculation is placed on the shaking table shaken cultivation, and frequency of oscillation is 150r/min.The shaking table room temperature is controlled at 24 ℃, and it is as clear as crystal to cultivate the visible culture fluid of 9d, and a large amount of Grifolas frondosa germ pompons are suspending in the liquid;
Second-class liquid isolate is made (liquid fermentation tank cultivation): culture fluid prescription, bottling, sterilization are with embodiment one, when being cooled to below 25 ℃ after the sterilization, insert the level liquid bacterial classification of 150ml, open inlet duct and constantly replenish the fresh air that has filtered, it is as clear as crystal to cultivate the visible culture fluid of 13d, a large amount of Grifolas frondosa germ pompons that suspending in the liquid are liquid spawn;
The grifola frondosus cultivating bacteria bag is made: the culture medium raw material proportioning is with embodiment one, and automatic packer packs, every packed wet feed 800g, high 14cm.The every bag of liquid spawn that meets 25ml in sterilization back places 23 ℃ of temperature, and relative moisture is 65%, CO
2Concentration is that 1000ppm cultivates, and the 30d mycelia is covered with the bacterium bag;
The fruiting cultivation management: bacterium bag scarfing mode is placed on 22 ℃ of temperature with embodiment one, relative moisture 98%, and the darkroom stimulates flower bud.The existing spherical original hase in 7d after-opening position.19 ℃ of temperature, relative moisture 93%, CO are set this moment
2Concentration 800ppm, indoor fluorescent lamp is opened.Spherical original hase is divided into the brain shape gradually, treats that fruit body length is fan-shaped to cap, can gather on the first appearance in the bacterium hole.
Compare with the cultural method that utilizes solid spawn, the present invention has following superiority:
Compare with solid spawn, the bacterial classification fabrication cycle of liquid spawn is short, and cell age is consistent and cost is low. Be distributed to the each several part of bacterium bag after the inoculation, mycelium pellet is sprouted simultaneously so that the bacterium time of sending out of bacterium bag is short and cell age is consistent, is convenient to the management of producing mushroom in later stage. Liquid spawn is paved with bacterium bag surface, and the bacterial classification fast-germination has significantly reduced bacterium bag pollution rate. Compare with using solid spawn, the former basic formation ratio height of the bacterium bag of liquid-spawn inoculation and grifola frondosus output height, quality are good.
Claims (2)
1. grifola frondosus liquid spawn is characterized in that preparing by the following method:
The female kind made: grifola frondosus mycelia piece is inserted the PDA medium, and 25 ℃ of dark culturing to mycelia are covered with the inclined-plane;
The level liquid bacterial classification is made: the culture fluid triangular flask of packing into, be cooled to after the sterilization below 25 ℃, and insert 3-5cm
2Mother's kind of smashing leaves standstill under 24-25 ℃ and cultivates 48h, places the shaking table shaken cultivation again, and frequency of oscillation is 100-150r/min, and the shaking table room temperature is cultivated 7-10d at 24-25 ℃;
Second-class liquid isolate is made: culture fluid is packed in the fermentation tank, and culture fluid is sterilized with the aeration equipment of fermentation tank, is cooled to after the sterilization below 25 ℃, inserts the level liquid bacterial classification, cultivates 12-15d, promptly obtains the grifola frondosus liquid spawn.
2. the method for the described grifola frondosus liquid strain cultivation of claim 1 grifola frondosus is characterized in that comprising the steps:
The grifola frondosus cultivating bacteria bag is made: the pack of medium wet feed, and every packed wet feed 700g-900g, high 12cm-14cm, sterilization, cooling, every bag of liquid spawn that meets 20-25ml places temperature 22-25 ℃, and relative moisture is 60%-70%, CO
2Concentration is to cultivate under the 1000-2000ppm condition, cultivates 25-30d;
The fruiting cultivation management: 10d afterripening after the mycelia purseful, treat that mycelia walks the back fully at lateral opening; Place temperature 22-24 ℃, relative moisture 95%-99%, the darkroom stimulates flower bud; The existing spherical original hase of 6-8d aperture position; Be provided with temperature 18-20 ℃, relative moisture 90%-95%, CO
2Concentration 500-800ppm, spherical original hase changes into the brain shape, treats that fruit body length is fan-shaped to cap, and can gather on the first appearance in the bacterium hole.
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|---|---|---|---|---|
| CN101946631A (en) * | 2010-08-12 | 2011-01-19 | 上海市农业科学院 | Bud pressing method for grifola frondosa |
| CN102432359A (en) * | 2010-09-29 | 2012-05-02 | 上海丰科生物科技股份有限公司 | Liquid medium for hypsizigus marmoreus liquid spawn culturing, preparation and cultivation method of liquid spawn |
| CN102498929A (en) * | 2011-10-12 | 2012-06-20 | 上海浦东天厨菇业有限公司 | Method for controlling yellow water exudation of plant bag-cultivated grifola frondosa at culture period |
| CN103155785A (en) * | 2011-12-09 | 2013-06-19 | 天津市金三农农业科技开发有限公司 | Establishment method for brown mushroom three-level strain propagation system |
| RU2485758C1 (en) * | 2012-02-07 | 2013-06-27 | Федеральное государственное бюджетное образовательное учреждение высшего профессионального образования "Национальный исследовательский Томский государственный университет" | METHOD OF GROWING MYCELIUM OF Grifola frondosa |
| CN103404364A (en) * | 2013-07-01 | 2013-11-27 | 鲁东大学 | Grifola frondosa liquid culture cultivating and high-yield cultivating method |
| CN103891527A (en) * | 2014-04-18 | 2014-07-02 | 湖南省春华生物科技有限公司 | Grifola frondosa factory production segmented mushroom cultivating method |
| CN104145711A (en) * | 2014-04-18 | 2014-11-19 | 湖南省春华生物科技有限公司 | Grifola frondosa three-stage culture production method |
| CN104285667A (en) * | 2014-09-10 | 2015-01-21 | 韦俊 | Cultivation method for improving polysaccharide content of grifola frondosa |
| CN104311348A (en) * | 2014-10-22 | 2015-01-28 | 常熟理工学院 | Improved liquid fermentation culture medium of pleurotus eryngii and method for culturing liquid strain of pleurotus eryngii by utilizing improved liquid fermentation culture medium |
| CN104541984A (en) * | 2015-01-28 | 2015-04-29 | 上海光明森源生物科技有限公司 | Culture compost for industrial production of grifola frondosa |
| CN105009929A (en) * | 2014-04-21 | 2015-11-04 | 浙江科技学院 | Grifold frondosa cultivation production technology with salvia miltiorrhiza dregs as main materials of cultivation bag |
| CN105638232A (en) * | 2014-11-13 | 2016-06-08 | 上海市农业科学院 | Domestic fungus liquid spawn culture method and medium thereof |
| CN106754409A (en) * | 2016-12-08 | 2017-05-31 | 段必儒 | White ginseng bacterium efficiently educates plant method |
| CN106906205A (en) * | 2017-01-10 | 2017-06-30 | 江苏大学 | The method for promoting grifola frondosus strain liquid state fermentation using magnetic field |
| CN107371804A (en) * | 2017-08-16 | 2017-11-24 | 吉首大学 | Utilize the method for giant knotweed cultivation grifola frondosus |
| CN107475135A (en) * | 2017-09-30 | 2017-12-15 | 山东省农业科学院农业资源与环境研究所 | A kind of special mother culture media of grifola frondosus and preparation method and application |
| CN111296177A (en) * | 2020-03-13 | 2020-06-19 | 江苏华绿生物科技股份有限公司 | Liquid strain culture medium applied to industrial cultivation of grifola frondosa and preparation method thereof |
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| CN101946631A (en) * | 2010-08-12 | 2011-01-19 | 上海市农业科学院 | Bud pressing method for grifola frondosa |
| CN102432359A (en) * | 2010-09-29 | 2012-05-02 | 上海丰科生物科技股份有限公司 | Liquid medium for hypsizigus marmoreus liquid spawn culturing, preparation and cultivation method of liquid spawn |
| CN102498929A (en) * | 2011-10-12 | 2012-06-20 | 上海浦东天厨菇业有限公司 | Method for controlling yellow water exudation of plant bag-cultivated grifola frondosa at culture period |
| CN103155785A (en) * | 2011-12-09 | 2013-06-19 | 天津市金三农农业科技开发有限公司 | Establishment method for brown mushroom three-level strain propagation system |
| CN103155785B (en) * | 2011-12-09 | 2014-04-16 | 天津市金三农农业科技开发有限公司 | Establishment method for brown mushroom three-level strain propagation system |
| RU2485758C1 (en) * | 2012-02-07 | 2013-06-27 | Федеральное государственное бюджетное образовательное учреждение высшего профессионального образования "Национальный исследовательский Томский государственный университет" | METHOD OF GROWING MYCELIUM OF Grifola frondosa |
| CN103404364A (en) * | 2013-07-01 | 2013-11-27 | 鲁东大学 | Grifola frondosa liquid culture cultivating and high-yield cultivating method |
| CN103891527A (en) * | 2014-04-18 | 2014-07-02 | 湖南省春华生物科技有限公司 | Grifola frondosa factory production segmented mushroom cultivating method |
| CN104145711A (en) * | 2014-04-18 | 2014-11-19 | 湖南省春华生物科技有限公司 | Grifola frondosa three-stage culture production method |
| CN104145711B (en) * | 2014-04-18 | 2016-03-02 | 湖南省春华生物科技有限公司 | A kind of grifola frondosus three-class strain preparation method |
| CN103891527B (en) * | 2014-04-18 | 2015-12-30 | 湖南省春华生物科技有限公司 | Mushroom method is educated in the segmentation of a kind of grifola frondosus factorial praluction |
| CN105009929A (en) * | 2014-04-21 | 2015-11-04 | 浙江科技学院 | Grifold frondosa cultivation production technology with salvia miltiorrhiza dregs as main materials of cultivation bag |
| CN104285667A (en) * | 2014-09-10 | 2015-01-21 | 韦俊 | Cultivation method for improving polysaccharide content of grifola frondosa |
| CN104311348A (en) * | 2014-10-22 | 2015-01-28 | 常熟理工学院 | Improved liquid fermentation culture medium of pleurotus eryngii and method for culturing liquid strain of pleurotus eryngii by utilizing improved liquid fermentation culture medium |
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| CN105638232A (en) * | 2014-11-13 | 2016-06-08 | 上海市农业科学院 | Domestic fungus liquid spawn culture method and medium thereof |
| CN104541984A (en) * | 2015-01-28 | 2015-04-29 | 上海光明森源生物科技有限公司 | Culture compost for industrial production of grifola frondosa |
| CN106754409A (en) * | 2016-12-08 | 2017-05-31 | 段必儒 | White ginseng bacterium efficiently educates plant method |
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| CN106906205A (en) * | 2017-01-10 | 2017-06-30 | 江苏大学 | The method for promoting grifola frondosus strain liquid state fermentation using magnetic field |
| CN107371804A (en) * | 2017-08-16 | 2017-11-24 | 吉首大学 | Utilize the method for giant knotweed cultivation grifola frondosus |
| CN107475135A (en) * | 2017-09-30 | 2017-12-15 | 山东省农业科学院农业资源与环境研究所 | A kind of special mother culture media of grifola frondosus and preparation method and application |
| CN111296177A (en) * | 2020-03-13 | 2020-06-19 | 江苏华绿生物科技股份有限公司 | Liquid strain culture medium applied to industrial cultivation of grifola frondosa and preparation method thereof |
| CN113812302A (en) * | 2021-10-11 | 2021-12-21 | 中国农业科学院农业资源与农业区划研究所 | Method for preparing liquid edible fungus strain |
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