CN112042471A - Industrialized production method of clitocybe maxima - Google Patents

Abstract

The invention provides an edible fungus mother culture medium or stock culture medium, a basic culture medium for edible fungus liquid culture and an industrialized production method of panus giganteus, wherein the method comprises the following steps: mother seed preparation, fungus bag preparation, liquid seed preparation and inoculation, spawn running management, fruiting management, harvesting and the like. The industrialized production method of the clitocybe maxima breaks through the mode that manual operation in the conventional production of the clitocybe maxima is used as a main mode, the land occupancy rate is high, the operation is complicated, the labor force is greatly saved, the labor efficiency is improved, the quality of the clitocybe maxima is improved, and the industrialized production method is suitable for industrialized production.

Description

Industrialized production method of clitocybe maxima

Technical Field

The invention relates to the field of edible mushroom cultivation, in particular to an industrialized production method of panus giganteus.

Background

China is a big country for producing edible fungi, more than 60 edible fungi can be artificially cultivated at present, and about 10 varieties can be cultivated in a large scale. In recent years, the edible fungus industry in China is developed vigorously, according to statistical investigation of China edible fungus Association on 27 provinces, autonomous regions and direct jurisdictions (provinces such as Tibet, Ningxia, Qinghai, Hainan, Hongkong and Macao and the like are not included), the total edible fungus yield in China in 2018 is 3842.04 ten thousand tons (fresh products, the same below), which is increased by 3.5% compared with 3712 ten thousand tons in 2017, the yield in 2018 is 2937.37 million yuan, and is increased by 7.92% compared with 2721.92 million yuan in 2017. The yield of 100 ten thousand tons is sequentially as follows: lentinus edodes (1043.12 ten thousand tons), Auricularia (674.03 ten thousand tons), Pleurotus ostreatus (642.82 ten thousand tons), Agaricus bisporus (307.49 ten thousand tons), Flammulina velutipes (257.56 ten thousand tons), Pleurotus eryngii (195.64 ten thousand tons) and Auricularia polytricha (189.85 ten thousand tons). The total yield of the first 7 varieties accounts for 86.15 percent of the total yield of edible fungi in China all the year round, and the edible fungi are the conventional main varieties for the edible fungi production in China.

In the last decade, with the flourishing domestic economy and the rapid improvement of the income level of residents, the industrial production can be rapidly increased from 8 ten thousand tons to 257 ten thousand tons in 2006 to 2016, the scale is increased by more than 32 times of the original scale, and the annual composite growth rate is about 41.5 percent. The daily yield of edible fungi produced by national edible fungi factory-scale enterprises in 2017 can reach 73225 tons, wherein the daily yield of flammulina velutipes is 3280 tons/day, the daily yield of pleurotus eryngii is 2705 tons/day, the daily yield of agaricus bisporus is 435 tons/day, the daily yield of hypsizygus marmoreus and white beech mushrooms is 318 tons/day, and the daily yield of hypsizygus marmoreus is 268 tons/day. In recent years, the daily productivity of these edible fungi has been greatly increased. The daily yield of the industrialized agaricus bisporus in 2017 reaches 435 tons, which is increased by 371 tons compared with 2010, and the annual compound speed increase is up to 31.6 percent.

Although the domestic edible fungus production is developed vigorously, the varieties are over concentrated, the yields of mushrooms, black fungus, oyster mushroom, agaricus bisporus, flammulina velutipes, pleurotus eryngii and auricularia polytricha ranked in the top seven account for 86.15% of the total yield of the domestic edible fungi all year round, the yields of mushrooms, black fungus and oyster mushroom ranked in the top three account for 61.42% of the total yield, and the industrial varieties mainly comprise flammulina velutipes, hypsizigus marmoreus, pleurotus eryngii, agaricus bisporus and the like; the production varieties are mainly medium-low temperature varieties, such as flammulina velutipes, the fruiting temperature of which is required to be about 4 ℃, and hypsizigus marmoreus, pleurotus eryngii, agaricus bisporus, shiitake mushrooms, agaric and the like are mainly at 15-20 ℃, so that the varieties suitable for medium-high temperature fruiting are lacked.

Panus giganteus (Berk.) Corner belongs to Basidiomycota, Basidiomycetes, Polyporales, Polyporaceae, and Dermatoloma (Panus), and the seed entity is recorded in China, Malaysia, Srilan, Indonesia, Vietnam, Laotian, Thailand, etc. singly or in a recurrent manner. The pork tripe mushroom is domesticated successfully by Sanming fungus research institute in Fujian province in China at first, and is named as the pork tripe mushroom due to unique taste and flavor and greasy taste like pork tripe.

The clitocybe maxima has higher nutritional value and edible value, the protein content of the clitocybe maxima is higher than that of common wood-rotting fungi such as shiitake mushroom and oyster mushroom and lower than that of straw-rotting fungi such as straw mushroom and agaricus bisporus, the fruiting body of the clitocybe maxima contains abundant carbohydrate, major elements such as potassium and phosphorus, trace elements such as copper, manganese, iron and zinc, essential amino acids such as threonine, valine, methionine, isoleucine, leucine, phenylalanine, lysine, histidine and arginine and abundant polysaccharide, and the research shows that the clitocybe maxima contains active ingredients such as oxidation resistance, fungus resistance, liver protection, inflammation resistance, diabetes resistance and tumor resistance.

The wild morchella is high in growing environment and suitable for being cultivated in areas with high temperature. With the continuous development of domestic edible fungus industry in recent years, the production scale of domestic edible fungi is larger and larger, and edible fungus products are more and more popular with consumers. The cultivation range of the clitocybe maxima as a medium-high temperature variety is gradually expanded from places such as Guangdong, Fujian and the like to places such as Zhejiang, Shandong, Beijing and the like, and the clitocybe maxima as a substitute mushroom variety in high-temperature seasons in summer is gradually attracted by edible mushroom producers.

Although artificial cultivation of clitocybe maxima is realized in China in many places, manual workshop type production is still the main point, and therefore, an industrial clitocybe maxima production process needs to be developed.

Disclosure of Invention

The invention firstly provides an edible fungus mother strain or stock strain culture medium, which comprises the following components in percentage by weight:

20-25% of potatoes; 2-3% of fructose; 3-5% of bran; 0-2% of peptone; 0-2% of beef extract; potassium dihydrogen phosphate 0.1-0.2%; magnesium sulfate 0.2-0.5%, VB₁ 0.1-0.3％；

It is characterized in that the green bean sprout extract also comprises green bean sprout leachate;

wherein the weight percentage content of the mung bean sprout leachate is 15-25%;

the preferable weight percentage content of the mung bean sprout leachate is 20 percent;

the further preferable edible fungus mother strain or stock strain culture medium comprises the following components in percentage by weight:

22.5 percent of potato, 2.5 percent of fructose, 4.0 percent of bran, 1 percent of peptone, 1 percent of beef extract, 0.15 percent of monopotassium phosphate, 0.35 percent of magnesium sulfate and VB₁0.2 percent of mung bean sprout leaching solution, and the balance of water;

the mung bean sprout leachate is prepared by the following method:

boiling semen Phaseoli Radiati Germinatus in water at 100 deg.C for 10-15min (weight ratio of semen Phaseoli Radiati Germinatus to water is 2: 5), filtering, and collecting filtrate to obtain the desired semen Phaseoli Radiati Germinatus lixivium.

The invention also provides a basic culture medium for liquid culture of edible fungi, which comprises the following components in percentage by weight:

2-8% of oak sawdust powder; 2-3% of fructose; 3-5% of bran; 0-2% of peptone; 0-2% of beef extract; potassium dihydrogen phosphate 0.1-0.2%; magnesium sulfate 0.2-0.5%, VB₁ 0.1-0.3％；

It is characterized in that the green bean sprout extract also comprises green bean sprout leachate;

wherein the weight percentage content of the mung bean sprout leachate is 15-25%;

the preferable weight percentage content of the mung bean sprout leachate is 20 percent;

the further preferable basic culture medium for liquid culture of the edible fungi comprises the following components in percentage by weight:

oak dust 5.0%, bran 4.0%, fructose 2.5%, peptone 1.0%, beef extract 1.0%, potassium dihydrogen phosphate 0.15%, magnesium sulfate 0.35%, VB₁0.2 percent of mung bean sprout leaching solution, and the balance of water;

the mung bean sprout leachate is prepared by the following method:

boiling semen Phaseoli Radiati Germinatus in water at 100 deg.C for 10-15min (weight ratio of semen Phaseoli Radiati Germinatus to water is 2: 5), filtering, and collecting filtrate to obtain the desired semen Phaseoli Radiati Germinatus lixivium.

The invention also provides an industrialized production method of the clitocybe maxima, which comprises the following steps:

(1) preparing a mother seed: the formula of the special mother culture medium for the morchella esculenta comprises the following components: 22.5 percent of potato, 2.5 percent of fructose, 4.0 percent of bran, 1.0 percent of peptone, 1.0 percent of beef extract, 0.15 percent of monopotassium phosphate, 0.35 percent of magnesium sulfate and VB₁0.2 percent of mung bean sprout leaching solution, and the balance of water;

(2) and (3) producing fungus bags: the production formula is as follows: 20% of oak sawdust, 20% of cotton seed hulls, 20% of corncobs, 20% of bagasse, 19% of bran and 1% of calcium carbonate. The water content is 65 percent, and the pH is natural; punching the middle of the bag when the amount of the packaged material is about 1Kg, covering the fungus bag, and sterilizing;

(3) liquid seed preparation and inoculation:

firstly, a first-stage shake flask seed culture medium is prepared and cultured according to the following formula: 200g of potato, 20g of fructose and VB₁2g, 1000mL of water and natural pH value;

secondly, preparing and culturing a second-stage shake flask seed culture medium according to the following formula:

the formula of the culture medium is as follows: 225g of potato, 25g of fructose, 40g of bran, 10g of peptone, 10g of beef extract, 1.25g of monopotassium phosphate, 3.5g of magnesium sulfate and VB₁2g, 200mL of mung bean sprout leachate, using distilled water to fix the volume to 1000mL, and enabling the pH value to be natural;

shaking the cultured first-stage shake flask seeds uniformly, transferring the seeds into a triangular flask with the capacity of 250mL and the capacity of 100mL of the culture medium by a liquid separator, wherein the inoculation amount of each flask is 10mL, and then placing the flask on a shaking table with the speed of 160r/min for carrying out constant-temperature shaking culture at 25 ℃ for 4 d;

thirdly, the liquid strain is subjected to basic culture according to the following formula, and the formula of the liquid strain basic culture medium is as follows: oak dust 5.0%, bran 4.0%, fructose 2.5%, peptone 1.0%, beef extract 1.0%, potassium dihydrogen phosphate 0.15%, magnesium sulfate 0.35%, VB₁0.2 percent of mung bean sprout leaching solution, and the balance of water;

after the liquid is prepared, inoculating fungus bags by using a GXJZ-Y type inoculating machine of a Xin edible fungus complete equipment company Limited in the Hongkong, and the inoculation amount of each bag is 10-15 mL;

(4) spawn running of the spawn package: the fungus bag adopts a high frame three-dimensional segmented culture mode: controlling the temperature in the culture room at 22-23 deg.C, the temperature in the fungus bag at 24-26 deg.C, and the humidity at 60-70% RH in the first 7 days, culturing in dark, and culturing in the presence of CO₂The concentration is less than 3000 ppm; controlling the temperature in the late culture room to be 20-21 ℃, the temperature in the fungus bag to be 22-23 ℃, the humidity to be 70-80% RH, culturing in the dark, and culturing in CO₂The concentration is less than 2000ppm, and the cultivation is continued for 30d after the hyphae are full, so that the hyphae are completely mature;

(5) and (3) fruiting management: standing the edge of the fungus bag after the fungus bag is mature, covering the fungus bag with turfy soil, and controlling the humidity of the covering soil to be within55-65%, the thickness of the covering soil is 3-4cm, the environmental temperature of the mushroom house is controlled at 23 ℃, the relative humidity is controlled at about 85%, and the illumination intensity is 200-₂The concentration is controlled below 3000 ppm; adjusting the ambient temperature of the mushroom house to 28 ℃ after three days, controlling the relative humidity to be more than 90 percent, and controlling the illumination intensity to be 200-₂Controlling the concentration to be below 2000ppm and maintaining for 24 hours; after 24h, the temperature of the mushroom house is reduced to 18 ℃, the relative humidity is controlled to be about 80 percent, and the illumination intensity is 500Lux and CO₂Controlling the concentration below 1500ppm, maintaining for 24 hr, allowing the primordium of Gaster Sus Domestica to break skin from the surface of the soil covering layer under the stimulation of high temperature and temperature difference, gradually growing into small bud, regulating the environmental temperature to above 23 deg.C-25 deg.C, not lower than 23 deg.C, controlling relative humidity at about 85%, and illuminating intensity at 500Lux and CO at about 85%₂The concentration is controlled below 2000 ppm; generally, the fruiting bodies can be harvested after the fruiting bodies are mature for 7-8 minutes after covering soil for about 12 days;

removing residual mushroom stem after harvesting fruiting body, adjusting temperature to 23 deg.C, controlling relative humidity at about 70%, keeping out of the sun, and keeping out of the sun with CO₂Controlling the concentration below 3000ppm to recover hypha for 7 d; and (7) stimulating fruiting again at high temperature after 7d, and harvesting the second tide of mushrooms.

The technical key points and the technical effects of the invention are as follows:

1. the mung bean sprout leachate is added into the seed culture medium and the liquid strain basic culture medium of the morchella esculenta, so that the morchella esculenta has faster mycelium setting value, strong and thick mycelium growth and stronger activity, and the culture time can be shortened by about 2 days.

2. The industrialized production of the clitocybe maxima is realized for the first time, the annual circulating production can be realized, the yield of the clitocybe maxima is increased, the content of heavy metal is effectively reduced, and the content of effective components is improved.

In a word, the industrial cultivation method of the clitocybe maxima breaks through the mode that manual operation in the existing production of the clitocybe maxima is used as a main mode, the land occupancy rate is high, the operation is complicated, labor force is greatly saved, the labor efficiency is improved, the content of heavy metal mercury, cadmium and chromium in the produced clitocybe maxima can be effectively reduced, the content of crude protein, total sugar and crude fiber in sporocarp is improved, the quality of the clitocybe maxima is improved, and the industrial cultivation method is suitable for industrial production.

Drawings

FIG. 1, two forms of bag (A: conventional semi-automatic bag-filling, B: automatic bag-filling machine of embodiment 3)

FIG. 2, two modes of strain morphology (A: conventional solid strain, B: liquid strain of example 3)

FIG. 3, two modes of culture in bags (A: conventional shelf culture, B: three-dimensional high-frame culture according to example 3)

FIG. 4 shows two modes of fruiting field (A: traditional buried soil-covered cultivation, B: industrial environmental control layer rack stereoscopic cultivation of example 3)

FIG. 5 shows two patterns of fruiting photos (A: fruiting in traditional buried soil covering mode, long period, large influence of soil quality, irregular fruiting and seasonal production; B: industrialized production, controllable environment, annual stereo production, influenced by soil environment and controllable quality)

FIG. 6 shows the development process of fruiting body in industrialized production of Gaster Sus Domestica

FIG. 7 shows the harvesting criteria of the fruiting body in the industrialized production of Gaster Sus Domestica

Detailed Description

The main raw material sources are as follows:

and (3) strains of the clitocybe maxima: shen mushroom No. 1 (Shanghai city academy of agricultural sciences edible fungus institute)

Mung bean sprouts, potatoes and bran are common commercial products;

fructose, peptone, beef extract, potassium dihydrogen phosphate, magnesium sulfate, VB₁: purchased from chemical reagents of national drug group, ltd;

oak dust and bran: purchased from Shanghai national Sen Biotech, Inc.;

EXAMPLE 1 stock culture Medium preparation

225g of potato, 25g of fructose, 40g of bran, 10g of peptone, 10g of beef extract, 1.25g of monopotassium phosphate, 3.5g of magnesium sulfate and VB₁2g, 200mL of mung bean sprout leachate, using distilled water to fix the volume to 1000mL, and enabling the pH value to be natural;

the mung bean sprout leachate is commercially available mung bean sprout, 200g of mung bean sprout is placed into 500mL of distilled water, boiled at 100 ℃ for 10min, filtered by using 4 layers of gauze, and the filtrate is collected for later use, and is boiled at present to prevent deterioration.

EXAMPLE 2 liquid Strain basal Medium preparation

The basic culture medium formula comprises: 5Kg of oak chip powder, 4Kg of bran, 2.5Kg of fructose, 1Kg of peptone, 1Kg of beef extract, 150g of monopotassium phosphate, 350g of magnesium sulfate and VB₁200g, 20L of mung bean sprout leachate, using distilled water to fix the volume to 100L, and keeping the pH natural; the formulation of the mungbean sprout leachate was as described in example 1.

Example 3 Industrial cultivation of Gaster Sus Domestica

And (3) strains of the clitocybe maxima: shiitake mushroom No. 1, institute of domestic fungus, academy of agricultural sciences, Shanghai city;

(1) preparing a mother seed: the mother culture medium formula special for the morchella esculenta in example 1 was used

(2) And (3) producing fungus bags: the production formula is as follows: 20% of oak sawdust, 20% of cotton seed hulls, 20% of corncobs, 20% of bagasse, 19% of bran and 1% of calcium carbonate. The water content is 65 percent, and the pH is natural. A fungus bag with the size of 17cm x 33cm is adopted, an automatic bag making machine is used for making bags, the amount of each bag is about 1Kg, a hole is punched in the middle, and the fungus bags are covered. Sterilizing in an autoclave at 121 ℃ for 2h, and cooling in a cooling chamber to room temperature;

wherein the main steps of the sterilization operation are as follows: introducing steam into a sterilizer chamber, wherein the temperature in a pot body reaches 98 ℃, and the time is about 30 min; pumping the air in the sterilizer chamber by a vacuum pump to reach a specified vacuum degree; thirdly, repeating the processes of the first step and the second step to reach the set times; introducing steam into the sterilizer chamber, heating the sterilized object to raise the temperature from 102 ℃ to 121 ℃, and keeping the temperature at the set temperature of 121 ℃ for 120min to achieve the aim of sterilization; stopping ventilation, reducing the temperature from 117 ℃ to 106 ℃ for 1 hour, discharging steam in a sterilization room: sixthly, vacuumizing by a vacuum pump, reducing the temperature from 106 ℃ to 103 ℃, refluxing air, and drying the sterilized culture bottles for 15 min; seventhly, repeating the process of the sixth step, and reducing the temperature from 103 ℃ to 98 ℃.

(3) Liquid seed preparation and inoculation:

the preparation process of the liquid seed comprises the following steps:

selecting soybean-sized hypha blocks from a mother seed test tube, inoculating the hypha blocks in the middle of a prepared PDA inclined plane, culturing at a constant temperature of 23 ℃ in the dark for 14 days, checking the existence of foreign bacteria pollution through microbiology, taking out the cultured hypha blocks, homogenizing by adding a proper amount of sterile water into a homogenizer, inoculating into a sterilized culture solution according to a ratio of 15% (V/V), fermenting and culturing to produce primary liquid seeds, homogenizing the primary liquid seeds, amplifying to produce secondary liquid seeds, homogenizing the secondary liquid seeds, and producing culture liquid seeds. Wherein the primary and secondary liquid seeds are produced by a triangular flask method.

First-stage shake flask seed culture medium preparation and culture

The formula of the culture medium is as follows: 200g of potato, 20g of fructose and VB₁2g, 1000mL of water and natural pH value.

Cutting the activated slant mother seeds into 1cm²Inoculating the hypha blocks into 250mL triangular flasks (6-8 hypha blocks are inoculated in each flask) containing 100mL of the culture medium, and placing the flasks on a shaking table at 160r/min at the constant temperature of 25 ℃ for shaking culture for 7d, wherein hypha grows vigorously to form a plurality of hypha balls.

② second-stage shake flask seed culture medium preparation and culture

The formula of the culture medium is as follows: (culture medium of example 1) Potato 225g, fructose 25g, bran 40g, peptone 10g, beef extract 10g, Potassium dihydrogen phosphate 1.25g, magnesium sulfate 3.5g, VB₁2g, 200mL of mung bean sprout leachate, using distilled water to fix the volume to 1000mL, and enabling the pH value to be natural;

shaking the cultured first-stage shake flask seeds uniformly, transferring into a triangular flask with a capacity of 250mL and 100mL of the culture medium by using a liquid separator, wherein the inoculation amount of each flask is 10mL, and then placing on a shaking table with the speed of 160r/min for shaking culture at the constant temperature of 25 ℃ for 4 days.

Preparation and culture of liquid strain culture medium

Basal medium formulation (medium of example 2): 5Kg of oak chip powder, 4Kg of bran, 2.5Kg of fructose, 1Kg of peptone, 1Kg of beef extract, 150g of monopotassium phosphate, 350g of magnesium sulfate and VB₁200g of mung bean sprout leachate, 20L of mung bean sprout leachate, and using distilled water to fix the volume to100L, and the pH is natural;

the preparation method comprises decocting semen Phaseoli Radiati Germinatus, placing testa Tritici and oak sawdust powder in the decoction, boiling and extracting for 10min, filtering with 8 layers of gauze, and collecting the extractive solution. And then adding the weighed other substances into the mixture respectively according to the experimental design. Inoculating according to the inoculation amount of 10%, controlling the culture temperature of liquid seeds at 24-25 deg.C for about 10 days, introducing purified air at the amount of 1: 0.5V/V.min for the first 3 days, and maintaining the pressure at 5-7 × 10⁴Pa, the ventilation was doubled at 7 d. After the culture is finished, the culture solution is clear and transparent, and a large amount of tiny uniform mycelium pellets are suspended in the culture solution without peculiar smell.

After the liquid is prepared, a GXJZ-Y type inoculation machine of a Xin edible fungus complete equipment company Limited in the Hongkong province is used for inoculation, and the inoculation amount of each bag is 10-15 mL.

(4) Spawn running of the spawn package: the fungus bag adopts a high frame three-dimensional segmented culture mode: controlling the temperature in the culture room at 22-23 deg.C, the temperature in the fungus bag at 24-26 deg.C, and the humidity at 60-70% RH in the first 7 days, culturing in dark, and culturing in the presence of CO₂The concentration is less than 3000 ppm; controlling the temperature in the late culture room to be 20-21 ℃, the temperature in the fungus bag to be 22-23 ℃, the humidity to be 70-80% RH, culturing in the dark, and culturing in CO₂The concentration is less than 2000ppm, and the culture is continued for 30 days after the hyphae are full, so that the hyphae are completely mature;

(5) and (3) fruiting management: standing the edge of the fungus bag after the fungus bag is mature, covering turfy soil in the fungus bag, controlling the soil covering humidity to be 55-65%, the soil covering thickness to be 3-4cm, controlling the ambient temperature of the mushroom house after soil covering to be 23 ℃, controlling the relative humidity to be about 85%, and controlling the illumination intensity to be 200-₂The concentration is controlled below 3000 ppm; adjusting the ambient temperature of the mushroom house to 28 ℃ after three days, controlling the relative humidity to be more than 90 percent, and controlling the illumination intensity to be 200-₂Controlling the concentration to be below 2000ppm and maintaining for 24 hours; after 24h, the temperature of the mushroom house is reduced to 18 ℃, the relative humidity is controlled to be about 80 percent, and the illumination intensity is 500Lux and CO₂Controlling the concentration below 1500ppm, maintaining for 24 hr, and stimulating at high temperature and temperature difference to allow the primordium of Gaster Sus Domestica to break skin from the surface of the soil covering layer and grow into small bud gradually, and regulating the environmental temperature to above 23-25 deg.CThe relative humidity is controlled to be about 85 percent at the temperature of lower than 23 ℃, the illumination intensity is 500Lux, and CO is₂The concentration is controlled below 2000 ppm; generally, the fruiting bodies can be harvested after the fruiting bodies are mature for 7-8 minutes after covering soil for about 12 days; removing residual mushroom stem after harvesting fruiting body, adjusting temperature to 23 deg.C, controlling relative humidity at about 70%, keeping out of the sun, and keeping out of the sun with CO₂Controlling the concentration below 3000ppm to recover hypha for 7 d; and (7) stimulating fruiting again at high temperature after 7d, and harvesting the second tide of mushrooms.

Comparative example: traditional manual production mode of panus giganteus (Wang Qingwu, Li Xiumei, Anxiong, lan Yu Fei. big clitocybe fungus bag cultivation high-yield technology, Shandong agricultural science, 2013, 45 (1): 130-132.)

And (3) strains of the clitocybe maxima: shiitake mushroom No. 1, institute of domestic fungus, academy of agricultural sciences, Shanghai city;

the production of the mushroom bag for producing the pork tripe mushroom adopts manual bagging, the sterilization adopts normal pressure sterilization, and the temperature of the normal pressure sterilization is maintained at 100 ℃ for more than 24 hours.

The strain uses solid strain, and is generally divided into three-stage seed production processes: mother seeds, stock seeds and cultivated seeds, wherein the production period of the mother seeds is 10d, the production period of the stock seeds is 30d, and the production period of the cultivated seeds is 30 d;

the mother seeds adopt the traditional PDA formula, and the original seed and cultivated species formula is as follows: 39% of each of the miscellaneous wood chips and the cotton seed hulls, 20% of the wheat bran and 1% of each of the sugar and the calcium carbonate (the weight percentage contents).

After hyphae grow over, the hyphae are manually inoculated into a cultivation bag, the inoculation amount is about 3 percent (volume ratio), the hyphae are placed into a cultivation room for spawn running cultivation after inoculation, the hyphae are cultivated in a dark place in the spawn running period, the temperature is kept between 25 and 30 percent, the air humidity is below 70 percent, ventilation is carried out for 2 times every day, about 2 hours every time, the indoor air is kept fresh, and the mixed fungus infection is prevented. The mycelia are grown over 30-40 days, and then continuously culturing for 30-60 days, and after the environmental temperature is kept above 23 ℃ and is proper, carrying out bag-removing, buried and earth-covering cultivation in a greenhouse.

Example 4

The morchella esculenta produced according to the method of example 3 and the method of comparative example was compared as follows:

1. the comparison of the basic production efficiency of the two cultivation processes is shown in table 1 below:

TABLE 1 comparison of basic production efficiencies for the two modes

As can be seen from table 1, when the method of example 3 of the present invention is used to cultivate panus giganteus, the production period is only 120d, annual production can be realized, the pollution rate is low, the quality is guaranteed, the environmental impact is less, and the bagging and inoculation are both mechanized, so the production efficiency is greatly improved.

2. The method for detecting the content of heavy metals in the clitocybe maxima fruiting bodies produced in two cultivation modes comprises the following steps: determination of total arsenic and inorganic arsenic in GB 5009.11-2014 food safety national standard food

Determination of lead in GB 5009.12-2017 national food safety standard food

Determination of cadmium in GB 5009.15-2014 food safety national standard food

Determination of total mercury and organic mercury in GB 5009.17-2014 food safety national standard food

Determination of chromium in GB 5009.123-2014 food safety national standard food

Specific detection comparison results are shown in table 2 below:

TABLE 2 comparison of heavy metal content in two patterns of fruiting bodies

	Comparative example	Example 3	National standard limit value (fresh product meter)
				Lead (mg/kg)	Not detected (<0.02)	Not detected (<0.02)	1.0
Arsenic (mg/kg)	0.048±0.005a	0.052±0.012a	0.5
				Mercury (mg/kg)	0.016±0.001a	0.011±0.002b	0.1
Cadmium (mg/kg)	0.212±0.001a	0.1905±0.0035b	0.2
				Chromium (mg/kg)	0.175±0.015a	0.045±0.013b	0.5

Note: a indicates that the difference is not statistically significant; b indicates that the difference is statistically significant. The limit value of edible fungi in GB2762-2017 pollutant limit in national food Standard for food safety is measured on fresh products, and is specified in the standard: in the case of a limit indicator that has a requirement for a product, wherein the limit of contaminants in the dry product is converted from the limit of contaminants in the corresponding fresh food product in combination with its dehydration rate or concentration rate. The dehydration rate or concentration rate can be determined by analysis of the food product, information provided by the producer, and other available data information, among other things. Unless otherwise specified.

As can be seen from Table 2, the fruiting body obtained by the cultivation method of example 3 of the present invention has significantly lower contents of mercury, cadmium and chromium than those of the comparative examples, especially much lower contents of chromium than those of the comparative examples. No lead was detected in the fruiting bodies obtained in both modes, and the arsenic content was slightly higher in example 3 than in the comparative example, but not statistically significant, i.e. there was no substantial difference.

3. The sporocarp obtained in the two cultivation modes is subjected to nutrient component detection, and specific data are shown in table 3. The detection methods of several nutrient components are respectively as follows:

determination of protein in crude protein GB 5009.5-2016 food safety national standard food

Determination of fat in crude fat GB 5009.6-2016 food safety national standard food

Determination of crude fiber in crude fiber GB/T5009.10-2003 plant food

Spectrophotometry for measuring total sugar content in red ginseng with total sugar NY/T2332-2013

Determination of content of crude polysaccharide in total polysaccharide NY/T1676-

TABLE 3 comparison of nutrient contents of fruiting bodies in two cultivation modes

	Comparative example	Example 3
			Crude protein content (g/kg)	311.81±2.63a	391.66±1.95b
Total polysaccharide content (mg/g)	22.61±0.076a	21.90±0.483a
			Total sugar (mg/g)	421.99±2.5831a	474.86±4.3427b
Crude fat (%)	4.68±0.1732a	9.36±0.1722b
			Crude fiber (%)	14.05±0.9683a	12.6±0.6615b

Note: a indicates that the difference is not statistically significant; b indicates that the difference is statistically significant.

As can be seen from Table 3, the fruiting bodies of example 3 have significantly higher contents of crude protein, total sugar and crude fat than the comparative examples, and significantly lower contents of crude fiber than the comparative examples, except that there is no statistical difference in total polysaccharides.

Claims (8)

1. An edible fungus mother strain or stock strain culture medium comprises the following components in percentage by weight:

20-25% of potatoes; 2-3% of fructose; 3-5% of bran; 0-2% of peptone; 0-2% of beef extract; potassium dihydrogen phosphate 0.1-0.2%; magnesium sulfate 0.2-0.5%, VB₁ 0.1-0.3％；

It is characterized in that the green bean sprout extract also comprises green bean sprout leachate;

wherein the weight percentage content of the mung bean sprout leachate is 15-25%.

2. The edible fungus mother culture or stock culture medium according to claim 1, wherein the weight percentage content of the mungbean sprout leachate is 20%.

3. The edible fungus mother culture or stock culture medium according to any one of claims 1 or 2, wherein the mungbean sprout leachate is prepared by the following method:

boiling mung bean sprouts in water at 100 ℃ for 10-15min, wherein the weight ratio of the mung bean sprouts to the water is 2: and 5, filtering, and collecting filtrate to obtain the required mung bean sprout leachate.

4. The edible fungus mother strain or stock strain culture medium according to any one of claims 1 or 2, which consists of the following components in percentage by weight:

22.5 percent of potato, 2.5 percent of fructose, 4.0 percent of bran, 1.0 percent of peptone, 1.0 percent of beef extract, 0.15 percent of monopotassium phosphate, 0.35 percent of magnesium sulfate and VB₁0.2 percent of mung bean sprout leaching solution, and the balance of water;

the mung bean sprout leachate is prepared by the following method:

boiling mung bean sprouts in water at 100 ℃ for 10-15min, wherein the weight ratio of the mung bean sprouts to the water is 2: and 5, filtering, and collecting filtrate to obtain the required mung bean sprout leachate.

5. A basic culture medium for liquid culture of edible fungi comprises the following components in percentage by weight:

2-8% of oak sawdust powder; 2-3% of fructose; 3-5% of bran; 0-2% of peptone; 0-2% of beef extract; potassium dihydrogen phosphate 0.1-0.2%; magnesium sulfate 0.2-0.5%, VB₁ 0.1-0.3％；

It is characterized in that the green bean sprout extract also comprises green bean sprout leachate;

wherein the weight percentage content of the mung bean sprout leachate is 15-25%.

6. The basal medium for liquid culture of edible fungi according to claim 5, wherein the weight percentage content of the leaching solution of mung bean sprouts is 20%.

7. The basal medium for liquid culture of edible fungi according to any one of claims 5 or 6, which consists of the following components in percentage by weight:

oak dust 5.0%, bran 4.0%, fructose 2.5%, peptone 1.0%, beef extract 1.0%, potassium dihydrogen phosphate 0.15%, magnesium sulfate 0.35%, VB₁0.2 percent of mung bean sprout leaching solution, and the balance of water;

the mung bean sprout leachate is prepared by the following method:

putting the mung bean sprouts into water, wherein the weight ratio of the mung bean sprouts to the water is 2: boiling at 5,100 deg.C for 10-15min, filtering, and collecting filtrate to obtain the desired mung bean sprout leachate.

8. An industrialized production method of panus giganteus is characterized by comprising the following steps:

(1) preparing a mother seed: the formula of the special mother culture medium for the morchella esculenta comprises the following components: 22.5 percent of potato, 2.5 percent of fructose, 4.0 percent of bran, 1 percent of peptone, 1 percent of beef extract, 0.15 percent of monopotassium phosphate, 0.35 percent of magnesium sulfate and VB₁0.2 percent of mung bean sprout leaching solution, and the balance of water;

(2) and (3) producing fungus bags: the production formula is as follows: 20% of oak sawdust, 20% of cotton seed hulls, 20% of corncobs, 20% of bagasse, 19% of bran and 1% of calcium carbonate. The water content is 65 percent, and the pH is natural; punching the middle of the bag when the amount of the packaged material is about 1Kg, covering the fungus bag, and sterilizing;

(3) liquid seed preparation and inoculation:

firstly, a first-stage shake flask seed culture medium is prepared and cultured according to the following formula: 200g of potato, 20g of fructose and VB₁2g, 1000mL of water and natural pH value;

secondly, preparing and culturing a second-stage shake flask seed culture medium according to the following formula:

the formula of the culture medium is as follows: 225g of potato, 25g of fructose, 40g of bran, 10g of peptone, 10g of beef extract, 1.25g of monopotassium phosphate, 3.5g of magnesium sulfate and VB₁2g, 200mL of mung bean sprout leachate, using distilled water to fix the volume to 1000mL, and enabling the pH value to be natural;

shaking the cultured first-stage shake flask seeds uniformly, transferring the seeds into a triangular flask with the capacity of 250mL and the capacity of 100mL of the culture medium by a liquid separator, wherein the inoculation amount of each flask is 10mL, and then placing the flask on a shaking table with the speed of 160r/min for carrying out constant-temperature shaking culture at 25 ℃ for 4 d;

thirdly, preparing a basic culture medium for the liquid strains according to the following formula: oak dust 5.0%, bran 4.0%, fructose 2.5%, peptone 1.0%, beef extract 1.0%, potassium dihydrogen phosphate 0.15%, magnesium sulfate 0.35%, VB₁0.2 percent of mung bean sprout leaching solution, and the balance of water;

after the liquid is prepared, inoculating fungus bags by using a GXJZ-Y type inoculating machine of a Xin edible fungus complete equipment company Limited in the Hongkong, and the inoculation amount of each bag is 10-15 mL;

(4) spawn running of the spawn package: the fungus bag adopts a high frame three-dimensional segmented culture mode: controlling the temperature in the culture room to be 22-23 ℃ in the first 7 days, controlling the temperature in the fungus bag to be 24-26 ℃ and the humidity to be 60-70% RH, culturing in the dark, and culturing in CO₂The concentration is less than 3000 ppm; controlling the temperature in the late culture room to be 20-21 ℃, the temperature in the fungus bag to be 22-23 ℃, the humidity to be 70-80% RH, culturing in the dark, and culturing in CO₂The concentration is less than 2000ppm, and the cultivation is continued for 30d after the hyphae are full, so that the hyphae are completely mature;

(5) and (3) fruiting management: standing the edge of the fungus bag after the fungus bag is mature, covering turfy soil in the fungus bag, controlling the soil covering humidity to be 55-65%, the soil covering thickness to be 3-4cm, controlling the ambient temperature of the mushroom house after soil covering to be 23 ℃, controlling the relative humidity to be about 85%, and controlling the illumination intensity to be 200-₂The concentration is controlled below 3000 ppm; adjusting the ambient temperature of the mushroom house to 28 ℃ after three days, controlling the relative humidity to be more than 90 percent, and controlling the illumination intensity to be 200-₂Controlling the concentration to be below 2000ppm and maintaining for 24 hours; after 24 hours, the temperature of the mushroom house is reduced to 18 DEG CRelative humidity is controlled to be about 80%, illumination intensity is 500Lux, and CO is₂Controlling the concentration below 1500ppm, maintaining for 24 hr, allowing the primordium of Gaster Sus Domestica to break skin from the surface of the soil covering layer under the stimulation of high temperature and temperature difference, gradually growing into small bud, regulating the environmental temperature to above 23 deg.C-25 deg.C, not lower than 23 deg.C, controlling relative humidity at about 85%, and illuminating intensity at 500Lux and CO at about 85%₂The concentration is controlled below 2000 ppm; generally, the fruiting bodies can be harvested after the fruiting bodies are mature for 7-8 minutes after covering soil for about 12 days;

removing residual mushroom stem after harvesting fruiting body, adjusting temperature to 23 deg.C, controlling relative humidity at about 70%, keeping out of the sun, and keeping out of the sun with CO₂Controlling the concentration below 3000ppm to recover hypha for 7 d; and (7) stimulating fruiting again at high temperature after 7d, and harvesting the second tide of mushrooms.

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