CN108330087B - Solid leaven for fermenting peanut straw - Google Patents
Solid leaven for fermenting peanut straw Download PDFInfo
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Abstract
The invention relates to a solid leaven for fermenting peanut straws, which can effectively solve the problems of inconvenient storage of liquid fermentation, easy aging of strains, loss of activity, great technical difficulty, unstable yield and large investment and solves the technical scheme that the solid leaven is prepared from five microorganisms, bran and corn flour, wherein the five microorganisms are trichoderma viride powder, phanerochaete chrysosporium powder, candida utilis powder, bacillus subtilis powder and enterococcus faecalis powder, and the effective viable count contained in each 1g of the solid leaven is 5 multiplied by 109‑20×109cfu, wherein each 1g of solid leaven contains 20-45% of bran and 10-30% of corn flour, and the raw materials are mixed together and uniformly stirred to obtain the peanut straw solid leaven.
Description
Technical Field
The invention belongs to the technical field of biology, and particularly relates to a solid leaven for fermenting peanut straws.
Background
The peanut is one of five important oil crops and is also one of important economic crops in China, Henan province is the first peanut production province in China, the planting area is more than 1500 mu of ten thousand, the total yield is more than 450 million tons, the yield of the peanut straw is 300-400 million tons, the yield is rich, the peanut straw is soft, the nutrient substances are rich, the crude protein is about 6% -20%, the crude fat is about 2% -3%, and the carbohydrate (mainly crude fiber) is about 46% -48%. Because peanut straws contain a large amount of cellulose, the digestive utilization rate of the peanut straws is low, the digestive absorption of the peanut straws by livestock is low, the production performance of the livestock is influenced, the contact of other nutrient substances and various enzymes is hindered, the digestive utilization rate of the feed is reduced, the application of the peanut straws in animal feed is limited, most of the peanut straws are directly burnt or decomposed and returned to the field, the resources are wasted, and the environment is seriously polluted. With the improvement of the living standard of people, the animal breeding industry is rapidly developed, the cultivated land area of China is gradually reduced, the grain yield is limited, and the contradiction that people and livestock fight for grain is increasingly prominent. Therefore, the peanut straws are subjected to certain biotechnology means and ways to convert cellulose in the peanut straws into monosaccharide, and the monosaccharide is used for producing protein feed, so that the palatability and the feed utilization rate of the peanut straws are changed, and the peanut straw protein feed has very important significance for relieving the shortage of grains and solving the problems of environmental pollution and the like.
At present, the process for fermenting crop straws mainly adopts liquid fermentation, but the liquid fermentation has the following defects: (1) the storage is inconvenient, and the strains are easy to age and lose the activity; (2) large technical difficulty (3) unstable yield; (4) the investment is large. The solid state fermentation has the advantages of low investment, low energy consumption, simplicity, convenience and the like, so the development of the peanut straw solid fermentation agent, the development and utilization of peanut straw resources by utilizing the microbial technology and the production of cheap animal high-protein feed have profound significance for forming grain-saving animal husbandry and promoting the sustainable development of agriculture and animal husbandry.
Disclosure of Invention
In view of the above situation, the present invention aims to provide a solid fermentation agent for peanut straw and a preparation method thereof, which can effectively solve the problems of inconvenient storage of liquid fermentation, easy aging and inactivation of strains, great technical difficulty, unstable yield and large investment.
The technical scheme includes that the solid leavening agent is prepared from five microorganisms, bran and corn flour, the five microorganisms are Trichoderma viride (Trichoderma viride) powder, Phanerochaete chrysosporium (Phanerochaete) powder, Candida utilis (Candida utilis) powder, Bacillus subtilis powder and Enterococcus faecalis (Enterococcus faecalis) powder, and the number of effective viable bacteria contained in each 1g of the solid leavening agent is 5 multiplied by 109-20×109The composite microbial inoculum comprises cfu, wherein the effective viable count of Trichoderma viride (Trichoderma viride) powder is 10-20%, the effective viable count of Phanerochaete chrysosporium (Phanerochaete) powder is 20-30%, the effective viable count of Candida utilis (Candida utilis) powder is 30-40%, the effective viable count of Bacillus subtilis (Bacillus subtilis) powder is 10-30%, the effective viable count of Enterococcus faecalis (Enterococcus faecalis) powder is 10-20%, each 1g of solid leavening agent contains 20-45% of bran and 10-30% of corn flour, and the composite microbial inoculum is obtained by mixing the raw materials together and uniformly stirring.
The trichoderma viride powder is prepared from the following components in parts by weight:
1) slant culture of test tube strains: inoculating Trichoderma viride to a PDA slant culture medium, and culturing at 28 deg.C for 5 days;
2) and (3) amplification culture: inoculating Trichoderma viride from PDA culture medium to Trichoderma viride amplification culture medium, and culturing at 28 deg.C for 5 days to obtain Trichoderma viride primary strain;
3) preparing a secondary seed liquid: inoculating the primary Trichoderma viride strain into a Trichoderma viride seed culture medium according to the inoculation amount of 5%, and continuously culturing at 30 ℃ and 200r/min for 24h to obtain a secondary Trichoderma viride strain;
4) fermentation in a fermentation tank: inoculating the secondary strain of the trichoderma viride into a trichoderma viride fermentation culture medium according to the inoculation amount of 5%, and continuously culturing for 28h at 30 ℃ and 240r/min to obtain trichoderma viride fermentation liquor;
5) centrifuging the Trichoderma viride fermentation liquor to obtain bacterial mud, adding 3% mannitol and 7% milk, and vacuum freeze drying to obtain Trichoderma viride powder with viable count of 3.2 × 1012cfu/g。
The phanerochaete chrysosporium powder is as follows:
1) slant culture of test tube strains: inoculating Phanerochaete chrysosporium to improved PDA slant culture medium, and culturing at 28 deg.C for 6 days;
2) and (3) amplification culture: inoculating Phanerochaete chrysosporium from PDA slant culture medium to Phanerochaete chrysosporium enlarged culture medium, and culturing at 30 deg.C for 3 days to obtain Phanerochaete chrysosporium first-stage strain;
3) preparing a secondary seed liquid: inoculating the first-level strain of Phanerochaete chrysosporium into a Phanerochaete chrysosporium seed culture medium according to the inoculation amount of 10%, and continuously culturing for 3 days at 30 ℃ and 200r/min to obtain a second-level strain of Phanerochaete chrysosporium;
4) fermentation in a fermentation tank: inoculating the phanerochaete chrysosporium secondary strain into a phanerochaete chrysosporium fermentation culture medium according to the inoculation amount of 10%, and continuously culturing for 3 days at 30 ℃ and 250r/min to obtain a phanerochaete chrysosporium fermentation liquid;
5) centrifuging the Phanerochaete chrysosporium fermentation liquid to obtain bacterial mud, adding 3% mannitol and 7% milk, and vacuum freeze drying to obtain Phanerochaete chrysosporium powder with viable count of 3 × 1011cfu/g。
The candida utilis strain powder is as follows:
1) slant culture of test tube strains: inoculating candida utilis to a PDA slant culture medium, and culturing for 24h at 28 ℃;
2) and (3) amplification culture: inoculating Candida utilis from PDA slant culture medium to Candida utilis enlarged culture medium, and culturing at 28 deg.C for 48 hr to obtain first-class yeast strain;
3) preparing a secondary seed liquid: inoculating the primary strain of the candida utilis into a candida utilis seed culture medium according to the inoculation amount of 5 percent, and continuously culturing for 48 hours at the temperature of 28 ℃ and at the speed of 200r/min to obtain a secondary strain of the candida utilis;
4) fermentation in a fermentation tank: inoculating the inoculation amount of the first-stage strain of the candida utilis into a candida utilis fermentation culture medium, and continuously culturing for 36 hours at the temperature of 30 ℃ and the speed of 240r/min to obtain candida utilis fermentation liquor;
5) centrifuging the Candida utilis fermentation liquor to obtain bacterial mud, adding 3% of mannitol and 7% of milk, and vacuum freeze drying to obtain Candida utilis powder with viable count of 4.7 × 109cfu/g。
The bacillus subtilis powder is as follows:
1) slant culture of test tube strains: inoculating the bacillus subtilis to a common agar slant culture medium and culturing for 18h at 37 ℃;
2) and (3) amplification culture: inoculating Bacillus subtilis from common agar slant culture medium to Bacillus subtilis enlarged culture medium, and culturing at 37 deg.C for 18 days to obtain Bacillus subtilis primary strain;
3) preparing a secondary seed liquid: inoculating the first-stage strain of the bacillus subtilis into a bacillus subtilis seed culture medium according to the inoculation amount of 8%, and continuously culturing for 12h at 35 ℃ and 180r/min to obtain a second-stage strain of the bacillus subtilis;
4) fermentation in a fermentation tank: inoculating the second-level strain of the bacillus subtilis into a bacillus subtilis fermentation culture medium according to the inoculation amount of 10%, and continuously culturing for 18h at 30 ℃ and 200r/min to obtain bacillus subtilis fermentation liquor;
5) centrifuging the Bacillus subtilis fermentation liquid to obtain bacterial mud, adding 5% mannitol and 5% milk, and vacuum freeze drying to obtain Bacillus subtilis powder with viable count of 4.3 × 1012cfu/g。
The enterococcus faecalis powder comprises the following components:
1) slant culture of test tube strains: inoculating enterococcus faecalis to a common agar slant culture medium, and culturing for 16h at 37 ℃;
2) and (3) amplification culture: taking enterococcus faecalis from a common agar slant culture medium, inoculating the enterococcus faecalis on a common broth culture medium, and culturing at 37 ℃ for 18h to obtain a first-level strain of the enterococcus faecalis;
3) preparing a secondary seed liquid: inoculating the first-stage enterococcus faecalis strain into an enterococcus faecalis seed culture medium according to the inoculation amount of 5%, and continuously culturing at 35 ℃ and 150r/min for 18h to obtain a second-stage enterococcus faecalis strain;
4) fermentation in a fermentation tank: inoculating the secondary strain of enterococcus faecalis to a enterococcus faecalis fermentation medium according to the inoculation amount of 5%, and continuously culturing at 35 ℃ and 180r/min for 16h to obtain an enterococcus faecalis fermentation liquid;
5) centrifuging the enterococcus faecalis fermentation liquid to obtain bacterial sludge, adding 3% mannitol and 7% milk, vacuum freeze drying to obtain enterococcus faecalis powder with viable count of 5.1 × 1010cfu/g。
The slant culture medium for PDA comprises peeling rhizoma Solani Tuber osi 200g, adding water 1000ml, boiling for 20min, filtering with gauze, weighing glucose 20g and agar 17g, adding into the filtrate, diluting with distilled water to 1L, and sterilizing at 121 deg.C for 30 min.
The improved PDA slant culture medium comprises: collecting peeled potato 200g, adding water 1000ml, boiling for 20min, filtering with gauze, weighing glucose 20g, agar 20g, and 3gKH2PO4、1.5gMgSO4·7H2O、0.1mgFeSO4·7H2O、0.2mgCuSO4·5H2O and 8mg of vitamin B1, and adding distilled water to a constant volume of 1L to obtain the final product.
The trichoderma viride amplification medium comprises: and (3) sufficiently and uniformly stirring 200mL of potato filtrate, 2g of glucose and 0.5g of urea, filling into a 250mL triangular flask, and sterilizing at 121 ℃ for 30min to obtain the potato beverage.
The Phanerochaete chrysosporium expanding culture medium comprises: 300mL of potato filtrate, 20g of glucose and 3gKH2PO4、1.5gMgSO4·7H2O、0.1mgFeSO4·7H2O、0.2mgCuSO4·5H2O and 8mg of vitamin B1, and adding distilled water to a constant volume of 1L to obtain the final product.
The candida utilis expanding culture medium comprises: collecting peeled fresh potato 200g, adding water 500mL, boiling for 20min, filtering with two layers of gauze, adding glucose 20g into the filtrate, heating to melt, diluting to 1L with distilled water, and sterilizing at 121 deg.C for 30 min.
The bacillus subtilis expanded culture medium comprises: adding 50mL of water into 5g of beef extract and 10g of peptone, heating until the beef extract and the peptone are completely dissolved, adding 500mL of water and 5g of NaCl to completely dissolve, adding distilled water to a constant volume of 1L, adjusting the pH value to 7.0 by using NaOH, and sterilizing at 121 ℃ for 30min to obtain the beef extract.
The Trichoderma viride fermentation medium (g/L) is as follows: 10g of peptone, 20g of glucose and 5g of beef extract, and the pH value is 6.8.
The Phanerochaete chrysosporium fermentation medium (g/L) is as follows: peptone 15g, glucose 25g, beef extract 3g, KH2PO45g、MgSO4·7H2O1g、FeSO4·7H2O0.15mg、CuSO4·5H2O0.2mg and vitamin B110mg, pH 7.0.
The Candida utilis fermentation medium (g/L) is as follows: 10g of beef extract, 9g of peptone, 20g of glucose, 5g of urea and MgSO4·7H2O1.5 g, pH 7.4.
The bacillus subtilis fermentation medium (g/L) is as follows: peptone 12g, beef extract 7g and sodium chloride 5, pH 7.3.
The enterococcus faecalis fermentation medium (g/L) is as follows: peptone 12g, beef extract 7g, glucose 10g and sodium chloride 3, and the pH value is 7.0.
The Trichoderma viride seed culture medium comprises: 200mL of potato filtrate, 20g of glucose, 5g (NH)4)2SO4And (5) adding distilled water to a constant volume of 1L to obtain the product.
The Phanerochaete chrysosporium seed culture medium comprises: 40g corn steep liquor, 20g glucose, 3gKH2PO4、2gMgSO4·7H2O、1g(NH4)2SO4And 50mg of vitamin B1, and adding distilled water to a constant volume of 1L to obtain the final product.
The candida utilis seed culture medium comprises the following components: 70g glucose, 5g (NH)4)2SO440g of peptone, sucrose, 2gKH2PO4And 2g of magnesium sulfate, and adding distilled water to a constant volume of 1L to obtain the magnesium sulfate.
The bacillus subtilis seed culture medium is as follows: and (3) adding 10g of yeast extract, 20g of peptone, 5g of NaCl, 10g of sucrose and 5g of agar powder, and diluting to a constant volume of 1L by using distilled water to obtain the yeast extract.
The enterococcus faecalis seed culture medium comprises: 10g of peptone, 3g of beef extract powder and 5g of sodium chloride, and adding distilled water to a constant volume of 1L.
The common agar slant culture medium comprises the following components: 10g of peptone, 3g of beef extract powder, 5g of sodium chloride and 15g of agar, and adding distilled water to a constant volume of 1L.
The general broth culture medium: 10g of peptone, 3g of beef extract powder and 5g of sodium chloride, and adding distilled water to a constant volume of 1L.
The peanut straw solid fermentation agent disclosed by the invention is formed by combining five active microorganisms, namely trichoderma viride powder, phanerochaete chrysosporium powder, candida utilis powder, bacillus subtilis powder and enterococcus faecalis, bran and corn flour, so that the content of crude protein in peanut straws can be increased, the content of cellulose can be reduced, the palatability and the feed utilization rate of the peanut straws can be changed, and the peanut straw solid fermentation agent is an innovation on the peanut straw fermentation agent.
Detailed Description
Example 1
The invention can be made from five microorganisms, bran and corn flour when in specific implementation, the five microorganisms are trichoderma viride powder, phanerochaete chrysosporium powder, candida utilis powder, bacillus subtilis powder and enterococcus faecalis powder, and the effective viable count contained in each 1g of solid leavening agent is 6 multiplied by 109cfu, wherein the effective viable count of the trichoderma viride powder accounts for 11 percent, the effective viable count of the phanerochaete chrysosporium powder accounts for 23 percent, the effective viable count of the candida utilis powder accounts for 31 percent, the effective viable count of the bacillus subtilis powder accounts for 24 percent, and the effective count of the enterococcus faecalis powderThe viable count accounts for 11%, and each 1g of the solid leaven contains 30% of bran and 30% of corn flour, and the raw materials are mixed together and stirred uniformly to obtain the biological leaven.
Example 2
The invention can be made from five microorganisms, bran and corn flour when in specific implementation, the five microorganisms are trichoderma viride powder, phanerochaete chrysosporium powder, candida utilis powder, bacillus subtilis powder and enterococcus faecalis powder, and the effective viable count contained in each 1g of solid leavening agent is 10 multiplied by 109The cfu comprises 15 percent of trichoderma viride powder effective viable bacteria, 25 percent of phanerochaete chrysosporium powder effective viable bacteria, 30 percent of candida utilis powder effective viable bacteria, 15 percent of bacillus subtilis powder effective viable bacteria, 15 percent of enterococcus faecalis powder effective viable bacteria, 20 percent of bran and 15 percent of corn flour in 1g of solid leavening agent, and the raw materials are mixed together and uniformly stirred to obtain the microbial fuel.
Example 3
The invention can be made from five microorganisms, bran and corn flour when in specific implementation, the five microorganisms are trichoderma viride powder, phanerochaete chrysosporium powder, candida utilis powder, bacillus subtilis powder and enterococcus faecalis powder, and the effective viable count contained in each 1g of solid leavening agent is 19 multiplied by 109The composite microbial agent comprises cfu, wherein the effective viable count of trichoderma viride powder is 17%, the effective viable count of phanerochaete chrysosporium powder is 21%, the effective viable count of candida utilis powder is 34%, the effective viable count of bacillus subtilis powder is 11%, the effective viable count of enterococcus faecalis powder is 17%, and each 1g of solid leavening agent contains 40% of bran and 10% of corn flour, and the raw materials are mixed together and stirred uniformly to obtain the composite microbial agent.
The five microorganisms show synergistic effect in the process of fermenting the peanut straws, the using effect is good, and the test data fully proves that the following is a solid leaven fermentation effect test of the invention:
test Material and Strain
Peanut straw: taking mildew-free dry peanut straws from a Hongrun family farm in the plain bridge area of Xinyang city, and processing the straws into powder by a feed grinder.
Strain: the peanut straw solid leaven prepared by the invention
The experimental site: xinyang agriculture and forestry college feed production laboratory;
2 instruments and apparatus
MBJ-80B incubator (Tianjin Sedris technologies, Inc.); YXQ-LS-100A pressure steam sterilizer (Jinan Xin Beixi Biotechnology Co., Ltd.); SW-CJ-2FD clean bench (air technologies, Inc., Antai, Suzhou); k1100 full-automatic Kjeldahl apparatus (Jinan ocean energy apparatus, Inc.); UV-1810 ultraviolet spectrophotometer (Beijing Pujingyo technologies, Ltd.).
3 method
3.1 pretreatment of peanut straw
Putting the prepared fermented feed into a 1 kg fermentation bottle, putting 0.7kg peanut straw in each bag, adding distilled water until the water content reaches 60%, wrapping with kraft paper, placing into a pressure cooker, sterilizing at 121 deg.C under high pressure for 30min, taking out, and placing on a clean bench to room temperature for use.
3.2 design of the experiment
The test is divided into 2 groups, namely a test group and a control group, the peanut straw of the control group is not added with fungi, the test group uses the leaven prepared by the invention, the inoculation amount is 10 percent, 5 times of each group are repeated, the peanut straw is crushed into 10 to 30 meshes, water is added to ensure that the water content of the peanut straw reaches 60 percent, 10 percent of peanut straw solid leaven and 1 percent of urea are added, and after being uniformly mixed, the mixture is fermented for 15 days at the constant temperature of 30 ℃.
3.3 Collection of samples
And (4) sampling at 15d of culture, and putting the taken sample into a constant temperature incubator to be dried at 40 ℃ for later use.
3.4 determination of nutrient composition
The determination of Crude Protein (CP), Crude Fiber (CF) and Crude Ash (CA) of the microbial fermentation straw is carried out according to GB/T6435-86, GB/T6432-86 and GB/T6434-86 standards.
4 results
The indexes of the main nutrient components of the leavening agent fermented peanut straw feed prepared by the invention are shown in table 1.
TABLE 1 Change in the Primary nutrient content after fermentation of peanut stalks
| Main nutrient components | Test group | Control group |
| Crude protein (%) | 14.73 | 11.51 |
| Coarse ash (%) | 9.39 | 11.78 |
| Crude fiber (%) | 28.49 | 41.30 |
| pH value | 4.2 | 7.1 |
The results show that after the peanut straws are subjected to the conversion action of the leavening agent, the crude protein is increased by 27.98%, the crude fiber is reduced by 31.02%, and the pH value is reduced by 2.9, which shows that when the peanut straws are fermented by the leavening agent, the crude fiber content of the peanut straws can be reduced, the crude protein of the peanut straws is improved, the digestion is promoted, the appetite is stimulated, and the utilization rate of the peanut straws is promoted.
The peanut straw solid fermentation agent disclosed by the invention is prepared by combining five active microorganisms, namely trichoderma viride powder, phanerochaete chrysosporium powder, candida utilis powder, bacillus subtilis powder and enterococcus faecalis, bran and corn flour, so that the content of crude protein in peanut straws can be increased, the content of cellulose can be reduced, the palatability and the feed utilization rate of the peanut straws can be changed, the peanut straw solid fermentation agent is an innovation on the peanut straw fermentation agent, the sustainable development of agriculture and animal husbandry is effectively promoted, and the peanut straw solid fermentation agent has great practical significance to economy and society.
Claims (6)
1. A solid leaven for fermenting peanut straws is characterized in that the solid leaven is prepared from five microorganisms, bran and corn flour, the five microorganisms are Trichoderma viride powder, Phanerochaete chrysosporium powder, Candida utilis powder, Bacillus subtilis powder and enterococcus faecalis powder, and the effective viable count of each 1g of the solid leaven is 5 multiplied by 109-20×109The composite microbial agent comprises cfu, wherein the effective viable count of trichoderma viride powder is 10-20%, the effective viable count of phanerochaete chrysosporium powder is 20-30%, the effective viable count of candida utilis powder is 30-40%, the effective viable count of bacillus subtilis powder is 10-30%, the effective viable count of enterococcus faecalis powder is 10-20%, each 1g of solid leavening agent contains 20-45% of bran and 10-30% of corn flour, and the raw materials are mixed together and stirred uniformly to obtain the composite microbial agent.
2. The solid fermentation agent for fermenting peanut straw as claimed in claim 1, which is prepared from five microorganisms, bran and corn flour, wherein the five microorganisms are trichoderma viride powder, phanerochaete chrysosporium powder, candida utilis powder, bacillus subtilis powder and enterococcus faecalis powder, and the effective viable count of each 1g of the solid fermentation agent is 6 x 109The cfu comprises 11 percent of effective viable count of trichoderma viride powder, 23 percent of effective viable count of phanerochaete chrysosporium powder, 31 percent of effective viable count of candida utilis powder, 24 percent of effective viable count of bacillus subtilis powder, 11 percent of effective viable count of enterococcus faecalis powder, 30 percent of bran and 30 percent of corn flour in 1g of solid leavening agent, and the raw materials are mixed togetherAnd uniformly stirring to obtain the product.
3. The solid fermentation agent for fermenting peanut straw as claimed in claim 1, which is prepared from five microorganisms, bran and corn flour, wherein the five microorganisms are trichoderma viride powder, phanerochaete chrysosporium powder, candida utilis powder, bacillus subtilis powder and enterococcus faecalis powder, and the effective viable count of each 1g of the solid fermentation agent is 10 x 109The cfu comprises 15 percent of trichoderma viride powder effective viable bacteria, 25 percent of phanerochaete chrysosporium powder effective viable bacteria, 30 percent of candida utilis powder effective viable bacteria, 15 percent of bacillus subtilis powder effective viable bacteria, 15 percent of enterococcus faecalis powder effective viable bacteria, 20 percent of bran and 15 percent of corn flour in 1g of solid leavening agent, and the raw materials are mixed together and uniformly stirred to obtain the microbial fuel.
4. The solid fermentation agent for fermenting peanut straw as claimed in claim 1, which is prepared from five microorganisms, bran and corn flour, wherein the five microorganisms are trichoderma viride powder, phanerochaete chrysosporium powder, candida utilis powder, bacillus subtilis powder and enterococcus faecalis powder, and the effective viable count of each 1g of the solid fermentation agent is 19 x 109The composite microbial agent comprises cfu, wherein the effective viable count of trichoderma viride powder is 17%, the effective viable count of phanerochaete chrysosporium powder is 21%, the effective viable count of candida utilis powder is 34%, the effective viable count of bacillus subtilis powder is 11%, the effective viable count of enterococcus faecalis powder is 17%, and each 1g of solid leavening agent contains 40% of bran and 10% of corn flour, and the raw materials are mixed together and stirred uniformly to obtain the composite microbial agent.
5. The solid leaven for fermenting peanut straw as claimed in any one of claims 1 or 2 to 4, wherein the Trichoderma viride powder is:
1) slant culture of test tube strains: inoculating Trichoderma viride to a PDA slant culture medium, and culturing at 28 deg.C for 5 days;
2) and (3) amplification culture: inoculating Trichoderma viride from PDA culture medium to Trichoderma viride amplification culture medium, and culturing at 28 deg.C for 5 days to obtain Trichoderma viride primary strain;
3) preparing a secondary seed liquid: inoculating the primary Trichoderma viride strain into a Trichoderma viride seed culture medium according to the inoculation amount of 5%, and continuously culturing at 30 ℃ and 200r/min for 24h to obtain a secondary Trichoderma viride strain;
4) fermentation in a fermentation tank: inoculating the secondary strain of the trichoderma viride into a trichoderma viride fermentation culture medium according to the inoculation amount of 5%, and continuously culturing for 28h at 30 ℃ and 240r/min to obtain trichoderma viride fermentation liquor;
5) centrifuging the Trichoderma viride fermentation liquor to obtain bacterial mud, adding 3% mannitol and 7% milk, and vacuum freeze drying to obtain Trichoderma viride powder with viable count of 3.2 × 1012cfu/g；
The phanerochaete chrysosporium powder is as follows:
1) slant culture of test tube strains: inoculating Phanerochaete chrysosporium to improved PDA slant culture medium, and culturing at 28 deg.C for 6 days;
2) and (3) amplification culture: inoculating Phanerochaete chrysosporium from PDA slant culture medium to Phanerochaete chrysosporium enlarged culture medium, and culturing at 30 deg.C for 3 days to obtain Phanerochaete chrysosporium first-stage strain;
3) preparing a secondary seed liquid: inoculating the first-level strain of Phanerochaete chrysosporium into a Phanerochaete chrysosporium seed culture medium according to the inoculation amount of 10%, and continuously culturing for 3 days at 30 ℃ and 200r/min to obtain a second-level strain of Phanerochaete chrysosporium;
4) fermentation in a fermentation tank: inoculating the phanerochaete chrysosporium secondary strain into a phanerochaete chrysosporium fermentation culture medium according to the inoculation amount of 10%, and continuously culturing for 3 days at 30 ℃ and 250r/min to obtain a phanerochaete chrysosporium fermentation liquid;
5) centrifuging the Phanerochaete chrysosporium fermentation liquid to obtain bacterial mud, adding 3% mannitol and 7% milk, and vacuum freeze drying to obtain Phanerochaete chrysosporium powder with viable count of 3 × 1011cfu/g；
The candida utilis strain powder is as follows:
1) slant culture of test tube strains: inoculating candida utilis to a PDA slant culture medium, and culturing for 24h at 28 ℃;
2) and (3) amplification culture: inoculating Candida utilis from PDA slant culture medium to Candida utilis enlarged culture medium, and culturing at 28 deg.C for 48 hr to obtain first-class yeast strain;
3) preparing a secondary seed liquid: inoculating the primary strain of the candida utilis into a candida utilis seed culture medium according to the inoculation amount of 5 percent, and continuously culturing for 48 hours at the temperature of 28 ℃ and at the speed of 200r/min to obtain a secondary strain of the candida utilis;
4) fermentation in a fermentation tank: inoculating the inoculation amount of the first-stage strain of the candida utilis into a candida utilis fermentation culture medium, and continuously culturing for 36 hours at the temperature of 30 ℃ and the speed of 240r/min to obtain candida utilis fermentation liquor;
5) centrifuging the Candida utilis fermentation liquor to obtain bacterial mud, adding 3% of mannitol and 7% of milk, and vacuum freeze drying to obtain Candida utilis powder with viable count of 4.7 × 109cfu/g；
The bacillus subtilis powder is as follows:
1) slant culture of test tube strains: inoculating the bacillus subtilis to a common agar slant culture medium and culturing for 18h at 37 ℃;
2) and (3) amplification culture: inoculating Bacillus subtilis from common agar slant culture medium to Bacillus subtilis enlarged culture medium, and culturing at 37 deg.C for 18 days to obtain Bacillus subtilis primary strain;
3) preparing a secondary seed liquid: inoculating the first-stage strain of the bacillus subtilis into a bacillus subtilis seed culture medium according to the inoculation amount of 8%, and continuously culturing for 12h at 35 ℃ and 180r/min to obtain a second-stage strain of the bacillus subtilis;
4) fermentation in a fermentation tank: inoculating the second-level strain of the bacillus subtilis into a bacillus subtilis fermentation culture medium according to the inoculation amount of 10%, and continuously culturing for 18h at 30 ℃ and 200r/min to obtain bacillus subtilis fermentation liquor;
5) centrifuging the Bacillus subtilis fermentation liquid to obtain bacterial sludge, adding 5% mannitol and 5% milk, vacuum freeze drying to obtain Bacillus subtilis powder with viable count of 4.31012cfu/g；
The enterococcus faecalis powder comprises the following components:
1) slant culture of test tube strains: inoculating enterococcus faecalis to a common agar slant culture medium, and culturing for 16h at 37 ℃;
2) and (3) amplification culture: taking enterococcus faecalis from a common agar slant culture medium, inoculating the enterococcus faecalis on a common broth culture medium, and culturing at 37 ℃ for 18h to obtain a first-level strain of the enterococcus faecalis;
3) preparing a secondary seed liquid: inoculating the first-stage enterococcus faecalis strain into an enterococcus faecalis seed culture medium according to the inoculation amount of 5%, and continuously culturing at 35 ℃ and 150r/min for 18h to obtain a second-stage enterococcus faecalis strain;
4) fermentation in a fermentation tank: inoculating the secondary strain of enterococcus faecalis to a enterococcus faecalis fermentation medium according to the inoculation amount of 5%, and continuously culturing at 35 ℃ and 180r/min for 16h to obtain an enterococcus faecalis fermentation liquid;
5) centrifuging the enterococcus faecalis fermentation liquid to obtain bacterial sludge, adding 3% mannitol and 7% milk, vacuum freeze drying to obtain enterococcus faecalis powder with viable count of 5.1 × 1010cfu/g。
6. The solid leaven for fermenting peanut straw as claimed in claim 5, wherein the PDA slant culture medium is prepared by taking 200g peeled potato, adding 1000ml water, boiling for 20min, filtering with gauze, weighing 20g glucose and 17g agar, adding into the filtrate, diluting with distilled water to 1L, and sterilizing at 121 deg.C for 30 min;
the improved PDA slant culture medium comprises: collecting peeled potato 200g, adding water 1000ml, boiling for 20min, filtering with gauze, weighing glucose 20g, agar 20g, and KH 3g2PO4、1.5g MgSO4·7H2O、0.1mg FeSO4·7H2O、0.2mg CuSO4·5H2O and 8mg of vitamin B1, and adding distilled water to a constant volume of 1L to obtain the final product;
the trichoderma viride amplification medium comprises: sufficiently stirring 200mL of potato filtrate, 2g of glucose and 0.5g of urea, putting into a 250mL triangular flask, and sterilizing at 121 ℃ for 30min to obtain the potato beverage;
the Phanerochaete chrysosporium expanding culture medium comprises: 300mL of potato filtrate, 20g of glucose and 3g of KH2PO4、1.5g MgSO4·7H2O、0.1mg FeSO4·7H2O、0.2mg CuSO4·5H2O and 8mg of vitamin B1, and adding distilled water to a constant volume of 1L to obtain the final product;
the candida utilis expanding culture medium comprises: collecting peeled fresh potato 200g, adding water 500mL, boiling for 20min, filtering with two layers of gauze, adding glucose 20g into the filtrate, heating to melt, adding water to constant volume of 1L, and sterilizing at 121 deg.C for 30min to obtain the final product;
the bacillus subtilis expanded culture medium comprises: adding 50mL of water into 5g of beef extract and 10g of peptone, heating until the beef extract and the peptone are completely dissolved, adding 500mL of water and 5g of NaCl to completely dissolve, adding water to a constant volume of 1L, adjusting the pH value to 7.0 by using NaOH, and sterilizing at 121 ℃ for 30min to obtain the beef extract;
the trichoderma viride fermentation medium comprises: 10g of peptone, 20g of glucose and 5g of beef extract, and the pH value is 6.8;
the Phanerochaete chrysosporium fermentation medium comprises: peptone 15g, glucose 25g, beef extract 3g, KH2PO4 5g、MgSO4·7H2O 1g、FeSO4·7H2O 0.15mg、CuSO4·5H2O0.2mg and vitamin B1 10mg, pH 7.0;
the candida utilis fermentation medium comprises: 10g of beef extract, 9g of peptone, 20g of glucose, 5g of urea and MgSO4·7H2O1.5 g, pH 7.4;
the bacillus subtilis fermentation medium comprises: peptone 12g, beef extract 7g and sodium chloride 5g, and the pH value is 7.3;
the enterococcus faecalis fermentation medium comprises: peptone 12g, beef extract 7g, glucose 10g and sodium chloride 3g, and the pH value is 7.0;
the Trichoderma viride seed culture medium comprises: 200mL of potato filtrate, 20g of glucose, 5g (NH)4)2SO4Adding distilled water to constant volume of 1L to obtain the final product;
the described chrysosporiumThe pangolin fungus seed culture medium comprises: 40g of corn steep liquor, 20g of glucose and 3g of KH2PO4、2g MgSO4·7H2O、1g (NH4)2SO4And 50mg of vitamin B1, and adding distilled water to a constant volume of 1L to obtain the final product;
the candida utilis seed culture medium comprises the following components: 70g glucose, 5g (NH)4)2SO440g of peptone, sucrose, 2g of KH2PO42g of magnesium sulfate, and fixing the volume to 1L by using distilled water to obtain the magnesium sulfate;
the bacillus subtilis seed culture medium is as follows: 10g of yeast extract, 20g of peptone, 5g of NaCl, 10g of sucrose and 5g of agar powder, and adding distilled water to a constant volume of 1L to obtain the yeast extract;
the enterococcus faecalis seed culture medium comprises: 10g of peptone, 3g of beef extract powder and 5g of sodium chloride, and adding distilled water to a constant volume of 1L;
the common agar slant culture medium comprises the following components: 10g of peptone, 3g of beef extract powder, 5g of sodium chloride and 15g of agar, and adding distilled water to a constant volume of 1L;
the general broth culture medium: 10g of peptone, 3g of beef extract powder and 5g of sodium chloride, and adding distilled water to a constant volume of 1L.
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