CN109258304A - A kind of production method and growth condition of hickory chick liquid culture bacteria - Google Patents

A kind of production method and growth condition of hickory chick liquid culture bacteria Download PDF

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Publication number
CN109258304A
CN109258304A CN201811159066.XA CN201811159066A CN109258304A CN 109258304 A CN109258304 A CN 109258304A CN 201811159066 A CN201811159066 A CN 201811159066A CN 109258304 A CN109258304 A CN 109258304A
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liquid
hickory chick
liquid culture
cultivation
morchella
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张津京
陈辉
王倩
汪虹
陈明杰
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Shanghai Academy of Agricultural Sciences
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Shanghai Academy of Agricultural Sciences
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    • AHUMAN NECESSITIES
    • A01AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
    • A01GHORTICULTURE; CULTIVATION OF VEGETABLES, FLOWERS, RICE, FRUIT, VINES, HOPS OR SEAWEED; FORESTRY; WATERING
    • A01G18/00Cultivation of mushrooms
    • A01G18/40Cultivation of spawn
    • AHUMAN NECESSITIES
    • A01AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
    • A01GHORTICULTURE; CULTIVATION OF VEGETABLES, FLOWERS, RICE, FRUIT, VINES, HOPS OR SEAWEED; FORESTRY; WATERING
    • A01G18/00Cultivation of mushrooms
    • A01G18/30Accessories for use before inoculation of spawn, e.g. sterilisers

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  • Life Sciences & Earth Sciences (AREA)
  • Mycology (AREA)
  • Environmental Sciences (AREA)
  • Micro-Organisms Or Cultivation Processes Thereof (AREA)
  • Apparatus Associated With Microorganisms And Enzymes (AREA)

Abstract

A kind of production method and growth condition of hickory chick liquid culture bacteria, this method breaks the mode of traditional solid-solid, using the mode of Liquid-liquid, cooperate the formula of our liquid cultivation seed culture medium simultaneously: sucrose 1.0-2.0% being added in the distilled water of 1L, albumen powder 0.10-0.50%, yeast powder 0.20-0.30%, bean cake powder 0.6-0.7%, K2HPO40.05-0.10%, KH2PO40.05-0.20% and MgSO40.05-0.10%, pH 6.3-6.8 are cultivated 5-7 days in 23 DEG C of fermentors, can carry out crop field or hayashishita sprinkling inoculation.On the one hand the process of production liquid cultivation seed is simple, at low cost and bacterium germination is fast, improves the efficiency of cultivation hickory chick significantly;On the other hand hickory chick cultivation under woods has successfully been carried out using liquid cultivation seed, has been that new cultivation mode is found in the cultivation of hickory chick.

Description

A kind of production method and growth condition of hickory chick liquid culture bacteria
Technical field
The present invention relates to the production method and growth condition of a kind of hickory chick liquid culture bacteria, belong to edible mashroom cultivating, Technical field of cultivation.
Background technique
Hickory chick (Morchella spp.) delicious meat is tender and crisp, nutritive and medicinal value is high, is a kind of food of preciousness Medicinal fungus, it is deep to be liked by various countries people, but wild toadstool yield is few, is not able to satisfy consumer demand, forest land is imitated wild Cultivation is to improve yield, indoors artificial cultivation is overcome to be not easy to form the cultural method of fructification.In production, solid is usually utilized Cultivar carries out inoculated and cultured, and the solid state cultivation kind amount needed is big and mycelia cell age difference is big, and the labour needed is also very Greatly, it is difficult to meet in production the needs of to cultivar amount.Therefore, explore the culture medium production method of liquid cultivation seed, and by this Technique Popularizing is applied in the pseudo-wild cultivating of hickory chick, is the effective way for capableing of large-scale production hickory chick.
Summary of the invention
It is an object of that present invention to provide the production methods and growth condition of a kind of hickory chick liquid culture bacteria.
To achieve the above object, present invention provide the technical scheme that a kind of production side of hickory chick liquid culture bacteria Method, including the following steps:
It is activated on the plate of PDA culture medium step 1: morchella mother culture is transferred to one and is accommodated, makes morchella mother culture Mycelia vitality restoration, then segmentation obtains hickory chick original seed PDA fungus block;
Step 2: the hickory chick original seed PDA fungus block is inoculated into the PDB fluid nutrient medium after sterilizing, in 20-25 DEG C of condition Under, it is cultivated 5-7 days with shaking table culture case in dark place, obtains hickory chick level-one kind;
Step 3: the hickory chick level-one kind is inoculated into the fermentor for accommodating hickory chick liquid cultivation seed culture medium, connect , by the fermentor after inoculation with 20-25 DEG C culture 5-7 days;The hickory chick liquid cultivation seed culture medium the preparation method comprises the following steps: It is added the sucrose of its quality 1.0-2.0% to 1 liter of distilled water, the yeast powder of the albumen powder of 0.10-0.50%, 0.20-0.30%, The bean cake powder of 0.6-0.7%, the K of 0.05-0.10%2HPO4, the KH of 0.05-0.20%2PO4With the MgSO of 0.05-0.10%4, mixing It is made after adjusting pH value to 6.3-6.8, then sterilized, cooling step after uniformly;
Step 4: being concentrated the hickory chick liquid spawn that third step culture obtains to get hickory chick using vacuum filtration machine Liquid culture bacteria.
Preferred technical solution are as follows: the condition of sterilizing are as follows: temperature is 121 DEG C, pressure 0.15Mpa, and sterilization time is 15min。
To achieve the above object, present invention provide the technical scheme that a kind of cultivation of hickory chick liquid culture bacteria is answered With, hickory chick liquid culture bacteria is placed in 4 DEG C of incubator, transport sowing place, by hickory chick liquid culture bacteria It is put into clean plastic water bucket, 9-11 times of quality of hickory chick liquid culture bacteria of commercially available drummed pure water, stirring is added It is uniformly dispersed in pure water to strain, directly crop field to be seeded or hayashishita is sprayed with water pump.
Detailed description of the invention
Attached drawing 1 is hickory chick liquid cultivation seed cultivation under woods hickory chick situation.
Since above-mentioned technical proposal is used, the present invention has the advantage, that compared with prior art
It is long that the present invention for traditional hickory chick strain solid connects the period in solid mode, and complicated for operation at high cost and cell age is poor The disadvantages of different big, is connect the production of hybrid seeds mode of liquid using liquid, and combines the formula of fluid nutrient medium, and a kind of hickory chick liquid is obtained The production method of cultivar is simultaneously carried out in hickory chick cultivation production.
Specific embodiment
Embodiments of the present invention are illustrated by particular specific embodiment below, those skilled in the art can be by this explanation Content disclosed by book is understood other advantages and efficacy of the present invention easily.
Embodiment one: a kind of production method and growth condition of hickory chick liquid culture bacteria
Hickory chick liquid cultivation seed carries out hickory chick cultivation in hayashishita
(1) preparation of hickory chick original seed
Morchella mother culture in test tube is transferred on PDA plate and is activated, continues switching activation 3 times in this way, to parent species Mycelia vitality restoration.
(2) preparation of hickory chick liquid level-one kind
100mLPDB fluid nutrient medium is added in the triangular flask of 250mL, is 121 DEG C in temperature, goes out under conditions of pressure 0.15Mpa Bacterium 15min.
(3) it by after sterilization and cooling, access diameter is 6 pieces of fungus block of hickory chick original seed PDA after the activation of 1cm, is placed in It cultivates 7 days the speed dark place of 23 DEG C of shaking table culture case 150rpm.
(4) culture medium prescription of hickory chick liquid cultivation seed is: the formula of fluid nutrient medium is: adding in the distilled water of 1L Enter to distill the sucrose of water quality 2.0%, 0.5% albumen powder, 0.3% yeast powder, 0.7% bean cake powder, 0.1% K2HPO4, 0.2% KH2PO4With 0.10% MgSO4, pH 6.8 is being 121 DEG C in temperature, sterilizing under conditions of pressure 0.15Mpa 20min, it is cooling.
(5) step (4) are housed by what the level liquid kind of step (2) cultured hickory chick was inoculated into that sterilizing cool down In the fermentor of hickory chick liquid cultivation seed.Then, the fermentor after inoculation is cultivated 7 days with 23 DEG C.
(6) after to the liquid cultivation seed culture of hickory chick, hickory chick liquid spawn is carried out using vacuum filtration machine Concentration, the hickory chick strain after concentration is placed in 4 DEG C of incubator, is transported sowing place, liquid cultivation seed is put into clean In net plastic water bucket, 10 times of commercially available drummed pure waters of quality of strain are added, stirring is uniformly dispersed in pure water to strain, Sprinkling hickory chick liquid cultivation seed directly is carried out in hayashishita to be seeded with water pump.Attached drawing 1 is the elite stand of hickory chick liquid culture Lower cultivation hickory chick situation, A-B: mycelium germination;C: former base is sprouted;D: fruit body development.
Embodiment two: a kind of production method and growth condition of hickory chick liquid culture bacteria
A kind of production method of hickory chick liquid culture bacteria, including the following steps:
It is activated on the plate of PDA culture medium step 1: morchella mother culture is transferred to one and is accommodated, makes morchella mother culture Mycelia vitality restoration, then segmentation obtains hickory chick original seed PDA fungus block;
Step 2: the hickory chick original seed PDA fungus block is inoculated into the PDB fluid nutrient medium after sterilizing, at 20 °C, use Shaking table culture case is cultivated 5 days in dark place, obtains hickory chick level-one kind;
Step 3: the hickory chick level-one kind is inoculated into the fermentor for accommodating hickory chick liquid cultivation seed culture medium, connect , the fermentor after inoculation is cultivated 5 days with 20 DEG C;The hickory chick liquid cultivation seed culture medium the preparation method comprises the following steps: to 1 liter Distilled water is added the sucrose of its quality 1.0%, 0.10% albumen powder, 0.20% yeast powder, 0.6% bean cake powder, 0.05% K2HPO4, 0.05% KH2PO4With 0.05% MgSO4, after adjusting pH value to 6.3, then sterilized, cooling step after mixing It is made;
Step 4: being concentrated the hickory chick liquid spawn that third step culture obtains to get hickory chick using vacuum filtration machine Liquid culture bacteria.
Preferred embodiment are as follows: the condition of sterilizing are as follows: temperature is 121 DEG C, pressure 0.15Mpa, and sterilization time is 15min。
Hickory chick liquid culture bacteria, is placed in 4 DEG C of incubator by a kind of growth condition of hickory chick liquid culture bacteria It is interior, sowing place is transported, hickory chick liquid culture bacteria is put into clean plastic water bucket, hickory chick liquid is added and plants The commercially available drummed pure water of 9 times of strain quality of training, stirring is uniformly dispersed in pure water to strain, with water pump directly to be seeded Crop field or hayashishita sprayed.
Embodiment three: a kind of production method and growth condition of hickory chick liquid culture bacteria
A kind of production method of hickory chick liquid culture bacteria, including the following steps:
It is activated on the plate of PDA culture medium step 1: morchella mother culture is transferred to one and is accommodated, makes morchella mother culture Mycelia vitality restoration, then segmentation obtains hickory chick original seed PDA fungus block;
Step 2: the hickory chick original seed PDA fungus block is inoculated into the PDB fluid nutrient medium after sterilizing, under the conditions of 25 DEG C, use Shaking table culture case is cultivated 6 days in dark place, obtains hickory chick level-one kind;
Step 3: the hickory chick level-one kind is inoculated into the fermentor for accommodating hickory chick liquid cultivation seed culture medium, connect , the fermentor after inoculation is cultivated 6 days with 25 DEG C;The hickory chick liquid cultivation seed culture medium the preparation method comprises the following steps: to 1 liter Distilled water is added the sucrose of its quality 1.5%, 0.3% albumen powder, 0.25% yeast powder, 0.65% bean cake powder, 0.07% K2HPO4, 0.12% KH2PO4With 0.07% MgSO4, after adjusting pH value to 6.5, then sterilized, cooling step after mixing It is made;
Step 4: being concentrated the hickory chick liquid spawn that third step culture obtains to get hickory chick using vacuum filtration machine Liquid culture bacteria.
Preferred embodiment are as follows: the condition of sterilizing are as follows: temperature is 121 DEG C, pressure 0.15Mpa, and sterilization time is 15min。
Hickory chick liquid culture bacteria, is placed in 4 DEG C of incubator by a kind of growth condition of hickory chick liquid culture bacteria It is interior, sowing place is transported, hickory chick liquid culture bacteria is put into clean plastic water bucket, hickory chick liquid is added and plants The commercially available drummed pure water of 11 times of strain quality of training, stirring is uniformly dispersed in pure water to strain, with water pump directly to waiting It is sprayed in the crop field of kind.
The above-described embodiments merely illustrate the principles and effects of the present invention, and is not intended to limit the present invention.It is any ripe The personage for knowing this technology all without departing from the spirit and scope of the present invention, carries out modifications and changes to above-described embodiment.Cause This, institute is complete without departing from the spirit and technical ideas disclosed in the present invention by those of ordinary skill in the art such as At all equivalent modifications or change, should be covered by the claims of the present invention.

Claims (3)

1.一种羊肚菌液体栽培菌种的制作方法,其特征在于:包括下列步骤:A method for preparing a liquid culture strain of Morchella, characterized in that the method comprises the following steps: 第一步:将羊肚菌母种转接到一容纳有PDA培养基的平板上进行活化,使羊肚菌母种的菌丝活力恢复,然后分割得到羊肚菌原种PDA菌块;The first step: transferring the Morchella mother to a plate containing PDA medium for activation, recovering the mycelial activity of the Morchella parent species, and then dividing to obtain the PDA strain of the morel mushroom; 第二步:将所述羊肚菌原种PDA菌块接种到灭菌后的PDB液体培养基,在20-25℃条件下,用摇床培养箱于暗处培养5-7天,得到羊肚菌一级种;The second step: inoculating the morel PDA bacteria into the sterilized PDB liquid medium, and incubating in a dark place for 5-7 days at 20-25 ° C to obtain a sheep. First-class species of the genus 第三步:将所述羊肚菌一级种接种到容纳有羊肚菌液体栽培种培养基的发酵罐中,接着,将接种后的发酵罐以20-25℃培养5-7天;所述羊肚菌液体栽培种培养基的制备方法为:向1升蒸馏水加入其质量1.0-2.0%的蔗糖,0.10-0.50%的蛋白粉、0.20-0.30%的酵母粉,0.6-0.7%的豆粕粉,0.05-0.10%的K2HPO4,0.05-0.20%的KH2PO4和0.05-0.10%的MgSO4,混合均匀后调节pH值至6.3-6.8,再经灭菌、冷却步骤后制得;The third step: inoculating the first species of the morel into a fermenter containing the liquid medium of the liquid culture of the morel, and then culturing the inoculated fermenter at 20-25 ° C for 5-7 days; The liquid culture medium for the Morchella is prepared by adding 1.0-2.0% sucrose, 0.10-0.50% protein powder, 0.20-0.30% yeast powder, 0.6-0.7% soybean meal to 1 liter of distilled water. Powder, 0.05-0.10% K 2 HPO 4 , 0.05-0.20% KH 2 PO 4 and 0.05-0.10% MgSO 4 , mix well and adjust the pH to 6.3-6.8, then sterilize and cool Have 第四步:利用真空抽滤机对第三步培养得到的羊肚菌液体菌种进行浓缩,即得羊肚菌液体栽培菌种。The fourth step: using a vacuum filter to concentrate the liquid strain of the morel obtained in the third step of cultivation, that is, the liquid culture strain of the morel. 2.根据权利要求1所述的羊肚菌液体栽培菌种的制作方法,其特征在于:灭菌的条件为:温度为121℃,压力为0.15Mpa,灭菌时间为15min。The method for preparing a liquid culture strain of Morchella according to claim 1, wherein the sterilization condition is: a temperature of 121 ° C, a pressure of 0.15 MPa, and a sterilization time of 15 min. 3.一种羊肚菌液体栽培菌种的栽培应用,其特征在于:将羊肚菌液体栽培菌种置于4℃的保温箱内,运输到播种地点,将羊肚菌液体栽培菌种放入洁净的塑料储水桶内,加入羊肚菌液体栽培菌种质量9-11倍的市售桶装纯净水,搅拌至菌种在纯净水中分散均匀,用水泵直接对待接种的大田或者林下进行喷洒。3. A cultivation application of a liquid culture strain of Morchella, characterized in that the liquid culture strain of Morchella is placed in an incubator at 4 ° C, transported to a seeding site, and the liquid culture strain of morel is placed. Into a clean plastic water storage tank, add 9-11 times of the quality of the liquid culture strain of Morchella, commercially available bottled pure water, stir until the strain is evenly dispersed in pure water, and directly treat the inoculated field or forest with a water pump. spray.
CN201811159066.XA 2018-09-30 2018-09-30 A kind of production method and growth condition of hickory chick liquid culture bacteria Pending CN109258304A (en)

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Cited By (5)

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CN111527820A (en) * 2020-06-15 2020-08-14 上海市农业科学院 Method for improving secondary salinized soil of vegetable greenhouse by using morchella cultivation
CN112772295A (en) * 2021-01-29 2021-05-11 贵州省生物研究所 Preparation method of morchella liquid strain and improved cultivar
CN115039635A (en) * 2022-06-08 2022-09-13 甘肃河州云菇农业科技有限公司 Liquid culture medium for preparing morchella stock culture in high altitude area and stock culture preparation method
CN115885770A (en) * 2022-11-23 2023-04-04 黑龙江省科学院微生物研究所 Morchella seeding method
CN118216369A (en) * 2024-04-24 2024-06-21 上海市农业科学院 Method for improving planting yield of Morchella strain by adding potassium salt

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Cited By (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN111527820A (en) * 2020-06-15 2020-08-14 上海市农业科学院 Method for improving secondary salinized soil of vegetable greenhouse by using morchella cultivation
CN112772295A (en) * 2021-01-29 2021-05-11 贵州省生物研究所 Preparation method of morchella liquid strain and improved cultivar
CN115039635A (en) * 2022-06-08 2022-09-13 甘肃河州云菇农业科技有限公司 Liquid culture medium for preparing morchella stock culture in high altitude area and stock culture preparation method
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CN118216369A (en) * 2024-04-24 2024-06-21 上海市农业科学院 Method for improving planting yield of Morchella strain by adding potassium salt

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