Kitchen waste liquid culture medium and fermentation culture method of trichoderma

Abstract

The invention relates to the technical field of biological fermentation, and particularly discloses a kitchen waste liquid culture medium and a fermentation culture method of trichoderma. The kitchen waste liquid culture medium provided by the invention is prepared from the following components: 5 to 10g / L of glucose, 0.05 to 0.1 g / L of ferrous sulfate, 0.5 to 1.0 g / L of magnesium sulfate and the balance of kitchen waste liquid, the content of organic matters in the kitchen waste liquid is 50-70 g / L, the content of nitrogen is 4-5 g / L, the content of phosphorus is 1-1.5 g / L, and the pH value is 4.5-7.0; the invention also provides a fermentation culture method of trichoderma. The fermentation culture method comprises the following steps: preparing a trichoderma spore bacterial suspension, performing liquid fermentation and performing solid fermentation, the kitchen waste liquid culture medium is adopted in the liquid fermentation. The kitchen waste liquid culture medium provided by the invention can be used for culturing trichoderma, so that the fermentation cost of the trichoderma is reduced, the fermentation efficiency of the trichoderma is improved, and large-scale industrial production of the trichoderma is realized.

Description

Technical Field

[0001] The present application relates to the field of biological fermentation technology, and in particular to a liquid culture medium of kitchen waste and a fermentation culture method of Trichoderma. Background Art

[0002] As an important rhizosphere growth-promoting fungus, Trichoderma has both protective and therapeutic effects in the biological control of plant pathologies. Its good biological control effect makes it an important development target for microbial agents and microbial fertilizers. At present, with the increasing awareness of ecological environmental protection and the development of microbial technology, research on the use of natural biological resources for biological control is becoming increasingly active, which not only helps to improve the disease resistance of crops, but also promotes the effective utilization of agricultural and forestry wastes and the development of a circular economy.

[0003] At present, a number of technologies have been reported for the industrial propagation of Trichoderma. For example, there are related technical reports that use wheat bran and sawdust as carriers for the fermentation of Trichoderma pseudokoningii; there are also technical reports that first use potato starch processing waste to propagate Trichoderma agents, and then use grass leaves and leaves for solid fermentation; there are also studies that use chemical synthetic culture media to amplify and propagate Trichoderma. Although the above methods have achieved large-scale cultivation of Trichoderma, some shortcomings have also been exposed in practical applications, especially the traditional methods, which mainly rely on agricultural and forestry waste as fermentation raw materials, resulting in general fermentation effects of Trichoderma and high fermentation costs; in addition, the above methods also have problems such as unstable and difficult to control raw material components, which directly affect the growth rate of Trichoderma and the product quality and output of its final bacterial agent.

[0004] Kitchen waste, also known as perishable waste, contains extremely high moisture and organic matter, is easy to rot and produce foul odors. As the amount of kitchen waste increases, the difficulty of disposal is also increasing, especially the incineration of solid phase waste, which will increase additional energy consumption and carbon emissions. Therefore, it is a good idea to process kitchen waste into a new culture medium for Trichoderma. On the one hand, it can improve the fermentation efficiency of Trichoderma and reduce the production cost of Trichoderma. On the other hand, it can solve the problem of kitchen waste resource utilization. Summary of the invention

[0005] In order to reduce the fermentation cost of Trichoderma, improve the fermentation efficiency of Trichoderma, and realize the resource utilization of kitchen waste, the present application provides a kitchen waste liquid culture medium and a fermentation culture method of Trichoderma.

[0006] In a first aspect, the present application provides a kitchen waste liquid culture medium, which adopts the following technical solution: A kitchen waste liquid culture medium, comprising the following components: 5-10 g / L glucose, 0.05-0.1 g / L ferrous sulfate, 0.5-1.0 g / L magnesium sulfate, and the remainder being kitchen waste liquid; The kitchen waste liquid has an organic matter content of 50-70 g / L, a nitrogen content of 4-5 g / L, a phosphorus content of 1-1.5 g / L, and a pH value of 4.5-7.0.

[0007] The present application uses kitchen waste liquid to prepare a kitchen waste liquid culture medium, which can efficiently promote the growth and reproduction of Trichoderma, significantly improve the biomass and activity of Trichoderma, and realize the industrial amplification and reproduction of Trichoderma. Kitchen waste liquid is the filtrate obtained after the treatment of kitchen waste. The kitchen waste liquid contains a large amount of organic matter, nitrogen, phosphorus and other nutrients, which can provide rich nutrients for the growth of Trichoderma, so that Trichoderma can rapidly proliferate on the culture medium and generate a large amount of mycelium and spores. At the same time, the glucose in the above-mentioned liquid culture medium can be used as a carbon source to effectively support the energy metabolism of Trichoderma and further promote its growth; ferrous sulfate and magnesium sulfate provide the trace elements required for the growth of Trichoderma, optimize the growth environment of Trichoderma, and enhance its cell viability. More noteworthy is that the liquid culture medium uses kitchen waste as the main raw material, which not only realizes the effective recycling of kitchen waste, reduces its impact on the environment, but also reduces the cost of fermentation culture medium, thereby contributing to the large-scale industrial production of Trichoderma, and at the same time provides a new idea for achieving green environmental protection and sustainable development.

[0008] Optionally, the method for preparing the kitchen waste liquid culture medium is: (1) subjecting the kitchen waste to impurity removal, pulping, wet heat treatment, solid-liquid separation, and oil-water separation in sequence, and obtaining a filtrate, which is the kitchen waste liquid; (2) Glucose, ferrous sulfate and magnesium sulfate are added to the kitchen waste liquid, mixed evenly, and sterilized at 121° C. for 0.5-3 h to obtain a kitchen waste liquid culture medium.

[0009] In some embodiments, the glucose content may be 5-8 g / L or 8-10 g / L.

[0010] In a specific embodiment, the glucose content may also be 5 g / L, 8 g / L or 10 g / L.

[0011] In some embodiments, the content of ferrous sulfate may be 0.05-0.08 g / L or 0.08-0.1 g / L.

[0012] In a specific embodiment, the content of ferrous sulfate may also be 0.05 g / L, 0.08 g / L or 0.1 g / L.

[0013] In some embodiments, the content of magnesium sulfate may be 0.5-0.8 g / L or 0.8-1 g / L.

[0014] In a specific embodiment, the content of magnesium sulfate may also be 0.5 g / L, 0.8 g / L or 1 g / L.

[0015] In a second aspect, the present application provides a fermentation culture method of Trichoderma, which adopts the kitchen waste liquid culture medium.

[0016] A fermentation and cultivation method of Trichoderma, comprising the following steps: preparing a suspension of Trichoderma spores, liquid fermentation and solid fermentation; Liquid fermentation: inoculating the suspension of Trichoderma spores into the liquid culture medium of kitchen waste, culturing at 25-28° C. and 170-200 rpm in a shaking table for 4-6 days to obtain Trichoderma liquid fermentation liquid; The concentration of Trichoderma spores in the Trichoderma spore suspension is 1×10 8 ~1×10 9 cfu / mL; the volume ratio of the Trichoderma spore suspension to the kitchen waste liquid culture medium is 1:(50-60).

[0017] The present application provides a method for fermenting and culturing Trichoderma, which significantly improves the reproduction efficiency and yield of Trichoderma by inoculating a suspension of Trichoderma spores into a liquid culture medium of kitchen waste according to the above-mentioned volume ratio and culturing under specific conditions, thereby obtaining an efficient liquid fermentation liquid of Trichoderma. The present application greatly improves the vitality of the Trichoderma strain and the production of its secondary metabolites by adjusting the fermentation temperature, fermentation time, the concentration of the suspension of Trichoderma spores, and the volume ratio of the suspension of Trichoderma spores to the liquid culture medium of kitchen waste, thereby improving the efficiency of the entire fermentation process, and ensuring the consistency and stability of the final inoculant product, providing technical support for the large-scale industrial production of Trichoderma.

[0018] In some embodiments, the weight ratio of the Trichoderma spore suspension to the liquid culture medium of kitchen waste can be 1:(40-50), 1:(40-55), 1:(40-60), 1:(40-70), 1:(50-55), 1:(50-60), 1:(50-70), 1:(55-60), 1:(55-70) or 1:(60-70).

[0019] In a specific embodiment, the weight ratio of the Trichoderma spore suspension to the kitchen waste liquid culture medium can also be 1:40, 1:50, 1:55, 1:60 or 1:70.

[0020] Optionally, the preparation method of the Trichoderma spore suspension is: inoculating Trichoderma spores into PDA culture medium, culturing at 28±2° C. for 4-5 days, alternating between 8-16 hours of light and 8-16 hours of darkness during the culturing process; after the culturing is completed, diluting with sterile water to obtain a Trichoderma spore suspension.

[0021] Optionally, the culture medium used for the solid fermentation is a kitchen waste solid culture medium; the kitchen waste solid culture medium comprises the following components in parts by weight: 60-80 parts of kitchen waste residue phase, 0-20 parts of sawdust and 20-40 parts of rice husk powder; the water content of the kitchen waste residue phase is 50-60%, and the pH value is 4.0-6.5.

[0022] The present application provides a novel solid culture medium for kitchen waste, which is prepared from the residue phase after kitchen waste treatment. The present application controls the addition amount of each component of the kitchen waste solid culture medium within the above range, so that the culture medium obtained has good air permeability and moisture retention ability, and can significantly improve the growth rate and mycelium density of Trichoderma, and effectively promote the accumulation of metabolites of Trichoderma during the solid fermentation process, thereby improving the yield and quality of Trichoderma preparations. Furthermore, since the kitchen waste residue phase contains rich organic matter and trace elements, the culture medium can also significantly promote the spore formation ability of Trichoderma, and enhance its adaptability to the environment and its ability to resist pathogens. In summary, the above-mentioned kitchen waste solid culture medium not only realizes the resource utilization of kitchen waste, but also significantly improves the industrial expansion efficiency and biological control effect of Trichoderma, and has a good prospect for use.

[0023] In the present application, the kitchen waste residue phase refers to the waste residue obtained by sequentially subjecting the kitchen waste to impurity removal, pulping, wet heat treatment, and solid-liquid separation.

[0024] Optionally, the kitchen waste solid culture medium comprises the following components in parts by weight: 60-80 parts of kitchen waste residue phase, 5-15 parts of sawdust and 25-35 parts of rice husk powder.

[0025] In some embodiments, the weight proportion of the kitchen waste residue phase may be 60-70 parts or 70-80 parts.

[0026] In a specific embodiment, the weight proportion of the kitchen waste residue phase may also be 60 parts, 70 parts or 80 parts.

[0027] In some embodiments, the sawdust may be present in an amount of 0-0.5 parts, 0.5-1.5 parts, or 1-1.5 parts by weight.

[0028] In a specific embodiment, the weight portion of the sawdust can also be 0 part, 0.5 part, 1 part or 1.5 parts.

[0029] In some embodiments, the sawdust may be present in an amount of 2-2.5 parts, 2-3.5 parts, 2-4 parts, 2.5-3.5 parts, 2.5-4 parts, or 3.5-4 parts by weight.

[0030] In a specific embodiment, the weight portion of the sawdust can also be 2 parts, 2.5 parts, 3.5 parts or 4 parts.

[0031] Optionally, the method for preparing the kitchen waste solid culture medium is: adding other components to the kitchen waste residue phase, sterilizing at 121° C. for 0.5-3 h, to obtain the kitchen waste solid culture medium.

[0032] Optionally, the specific steps of the solid fermentation are: inoculating the liquid fermentation liquid of Trichoderma into the solid culture medium of kitchen waste, and performing static shallow tray fermentation for 4-7 days at a temperature of 25-30°C and a humidity of 60-70%, wherein the thickness of the shallow tray fermentation is 2-5 cm, to obtain Trichoderma.

[0033] Optionally, the weight ratio of the Trichoderma liquid fermentation broth to the kitchen waste solid culture medium is 1:(5-10).

[0034] In some embodiments, the weight ratio of the Trichoderma liquid fermentation broth to the kitchen waste solid culture medium can be 1:(5-10) or 1:(10-15).

[0035] In a specific embodiment, the weight ratio of the Trichoderma liquid fermentation broth to the kitchen waste solid culture medium can also be 1:5, 1:8, 1:10 or 1:15.

[0036] In summary, this application has the following beneficial effects: 1. The present application provides a liquid culture medium of kitchen waste for Trichoderma fermentation, which can efficiently promote the growth and reproduction of Trichoderma, significantly increase the biomass and activity of Trichoderma, and realize the industrialized amplification and reproduction of Trichoderma; in addition, the above-mentioned liquid culture medium of kitchen waste uses kitchen waste as the main raw material, which not only realizes the effective recycling of kitchen waste and reduces its impact on the environment, but also reduces the cost of fermentation culture medium, which is conducive to the large-scale industrial production of Trichoderma.

[0037] 2. The present application also provides a kitchen waste solid culture medium, which is prepared from the residue phase after kitchen waste treatment. The kitchen waste solid culture medium has good air permeability and moisture retention ability, and can significantly increase the growth rate and mycelium density of Trichoderma, effectively promote the accumulation of metabolic products of Trichoderma, and improve the yield and quality of Trichoderma preparations.

[0038] 3. The present application further controls the volume ratio of the Trichoderma spore suspension to the liquid culture medium of kitchen waste within the range of 1: (50-60), and the weight ratio of the Trichoderma liquid fermentation liquid to the solid culture medium of kitchen waste within the range of 1: (5-10), which can significantly improve the fermentation efficiency of Trichoderma and make the reproduction yield of Trichoderma higher. DETAILED DESCRIPTION

[0039] In a first aspect, the present application provides a kitchen waste liquid culture medium, comprising the following components: 5-10 g / L glucose, 0.05-0.1 g / L ferrous sulfate, 0.5-1.0 g / L magnesium sulfate, and the remainder being kitchen waste liquid; The kitchen waste liquid has an organic matter content of 50-70 g / L, a nitrogen content of 4-5 g / L, a phosphorus content of 1-1.5 g / L, and a pH value of 4.5-7.0.

[0040] The preparation method of the kitchen waste liquid culture medium is: (1) subjecting the kitchen waste to impurity removal, pulping, wet heat treatment, solid-liquid separation, and oil-water separation in sequence, and obtaining a filtrate, which is the kitchen waste liquid; (2) Glucose, ferrous sulfate and magnesium sulfate are added to the kitchen waste liquid, mixed evenly, and sterilized at 121° C. for 0.5-3 h to obtain a kitchen waste liquid culture medium.

[0041] In a second aspect, the present application provides a kitchen waste solid culture medium, comprising the following components in parts by weight: 60-80 parts of kitchen waste residue phase, 0-20 parts of sawdust, and 20-40 parts of rice husk powder; the water content of the kitchen waste residue phase is 50-60%, and the pH value is 4.0-6.5. Further, the kitchen waste solid culture medium comprises the following components in parts by weight: 60-80 parts of kitchen waste residue phase, 5-15 parts of sawdust, and 25-35 parts of rice husk powder.

[0042] The method for preparing the kitchen waste solid culture medium comprises: adding sawdust and rice husk powder to the kitchen waste residue phase, mixing evenly, and sterilizing at 121° C. for 0.5-3 hours to obtain the kitchen waste solid culture medium.

[0043] In a third aspect, the present application provides a fermentation culture method of Trichoderma, comprising the following steps: (1) Preparation of Trichoderma spore suspension: inoculate Trichoderma spores into PDA medium and culture at 28±2°C for 4-5 days, with alternating light exposure for 8-16 h and darkness for 8-16 h during the culture process; after the culture is completed, dilute with sterile water to obtain a concentration of 1×10 8 ~1×10 9 cfu / mL of Trichoderma spore suspension.

[0044] (2) Liquid fermentation: inoculating the Trichoderma spore suspension into the kitchen waste liquid culture medium, culturing at 25-28° C. and 170-200 rpm on a shaker for 4-6 days to obtain a Trichoderma liquid fermentation liquid; the volume ratio of the Trichoderma spore suspension to the kitchen waste liquid culture medium is 1:(50-60).

[0045] (3) Solid fermentation: inoculating the liquid fermentation liquid of Trichoderma into the solid culture medium of kitchen waste, and fermenting in a static shallow tray for 4-7 days at a temperature of 25-30° C. and a humidity of 60-70%; the thickness of the shallow tray fermentation is 2-5 cm, and Trichoderma is obtained; the weight ratio of the liquid fermentation liquid of Trichoderma to the solid culture medium of kitchen waste is 1:(5-10).

[0046] In the examples of the present application, kitchen waste liquid and kitchen waste residue phase were obtained from Shanghai Wenxin Biotechnology Co., Ltd. The organic matter content of the kitchen waste liquid was 62 g / L, the nitrogen content was 4.8 g / L, the phosphorus content was 1.3 g / L, and the pH value was 6.1; the organic matter content of the kitchen waste residue phase was 75%, the pH value was 5.3, the nitrogen content was 4.2%, and the water content was 55%; the preservation number of the Trichoderma harzianum strain was CICC13056, which was purchased from the China Industrial Microbiological Culture Collection Management Center; sawdust was purchased from Caoxian Shenhang Wood Industry Co., Ltd.; rice husk powder was purchased from Shijiazhuang Lvwan Agricultural and Sideline Products Co., Ltd.; the raw materials, reagents, solvents, etc. of this application can all be obtained commercially.

[0047] The present application is further described in detail below in combination with preparation examples, embodiments, and performance testing experiments.

[0048] Preparation Example 1-7 Preparation Example 1-7 provides a kitchen waste liquid culture medium.

[0049] The difference between the above preparation examples is that the content of each component in the kitchen waste liquid culture medium is specifically shown in Table 1 below.

[0050] The preparation method of the kitchen waste liquid culture medium provided in Preparation Example 1-7 comprises the following steps: adding glucose, ferrous sulfate and magnesium sulfate to 1L of kitchen waste liquid, mixing evenly, sterilizing at 121° C. for 2 hours, and obtaining a kitchen waste liquid culture medium.

[0051] Table 1 Content of each component in the kitchen waste liquid culture medium of Preparation Examples 1-7 Preparation Example 8-17 Preparation Example 8-17 provides a kitchen waste solid culture medium.

[0052] The difference between the above preparation examples is that the content of each component in the kitchen waste solid culture medium is specifically shown in Table 2 below.

[0053] The preparation method of the kitchen waste solid culture medium provided in Preparation Example 8-17 comprises the following steps: adding sawdust and rice husk powder to the kitchen waste residue phase, mixing evenly, sterilizing at 121° C. for 2 hours, and obtaining the kitchen waste solid culture medium.

[0054] Table 2 Content of each component in the kitchen waste solid culture medium of Preparation Examples 8-17 Examples 1-16 Embodiments 1-16 respectively provide a fermentation culture method of Trichoderma.

[0055] The difference between the above embodiments is that the sources of the kitchen waste liquid culture medium and the kitchen waste solid culture medium are specifically shown in Table 3 below.

[0056] The fermentation culture method of Trichoderma provided in Examples 1-16 comprises the following steps: (1) Preparation of Trichoderma spore suspension: inoculate Trichoderma harzianum spores into PDA medium and culture at 28±2°C for 4-5 days, with 12 h of light and 12 h of darkness alternating during the culture process; after the culture is completed, dilute with sterile water to obtain a concentration of 1×10 8 cfu / mL of Trichoderma spore suspension.

[0057] (2) Liquid fermentation: inoculating the Trichoderma spore suspension into the kitchen waste liquid culture medium, culturing at 27° C. and 180 rpm in a shaking incubator for 5 days to obtain a Trichoderma liquid fermentation liquid; the volume ratio of the Trichoderma spore suspension to the kitchen waste liquid culture medium is 1:55.

[0058] (3) Solid fermentation: inoculating the liquid fermentation liquid of Trichoderma into the solid culture medium of kitchen waste, and fermenting in a static shallow tray for 6 days at a temperature of 27° C. and a humidity of 65%; the thickness of the shallow tray fermentation is 4 cm, and obtaining Trichoderma; the weight ratio of the liquid fermentation liquid of Trichoderma to the solid culture medium of kitchen waste is 1:8.

[0059] Table 3 Sources of kitchen waste liquid culture medium and kitchen waste solid culture medium in Examples 1-16 Examples 17-20 Examples 17-20 respectively provide a fermentation culture method of Trichoderma.

[0060] The difference between the above embodiment and embodiment 2 is that the volume ratio of the Trichoderma spore suspension to the kitchen waste liquid culture medium is specifically shown in Table 4 below.

[0061] Table 4 Volume ratio of Trichoderma spore suspension to kitchen waste liquid culture medium in Examples 17-20 Example Volume ratio of Trichoderma spore suspension to liquid culture medium of kitchen waste 2 1：55 17 1：40 18 1：50 19 1：60 20 1：70 Examples 21-23 Examples 21-23 respectively provide a fermentation culture method of Trichoderma.

[0062] The difference between the above embodiment and embodiment 2 is that the weight ratio of the liquid fermentation liquid of Trichoderma to the solid culture medium of kitchen waste is specifically shown in Table 5 below.

[0063] Table 5 Weight ratio of Trichoderma liquid fermentation broth to kitchen waste solid culture medium in Examples 21-23 Example Weight ratio of liquid fermentation liquid of Trichoderma to solid culture medium of kitchen waste 2 1：8 21 1：5 22 1：10 23 1：15 Comparative Example 1 Comparative Example 1 provides a fermentation culture method of Trichoderma.

[0064] The difference between the comparative example and Example 2 is that the kitchen waste liquid culture medium is replaced by a liquid Sabouraud culture medium, whose formula is: peptone 10 g / L, glucose 40 g / L.

[0065] Comparative Example 2 Comparative Example 2 provides a fermentation culture method of Trichoderma.

[0066] The difference between the above comparative example and Example 2 is that in the kitchen waste liquid culture medium of Comparative Example 2, the organic matter content in the kitchen waste liquid is 73 g / L, the nitrogen content is 3.7 g / L, and the phosphorus content is 2.3 g / L.

[0067] Comparative Example 3 Comparative Example 3 provides a fermentation culture method of Trichoderma.

[0068] The difference between the above comparative example and Example 2 is that: Comparative Example 3 directly uses kitchen waste liquid sterilized at 121° C. for 2 hours as the kitchen waste liquid culture medium.

[0069] Performance testing The Trichoderma harzianum strains were fermented and cultured using the Trichoderma fermentation culture methods provided in Examples 1-23 and Comparative Examples 1-3, and the spore production was counted by the hemocytometer counting method. The results are shown in Table 6 below.

[0070] Table 6 Spore production test results of Examples 1-23 and Comparative Examples 1-3 According to the test results in Table 6, the fermentation and cultivation methods of Trichoderma provided in Examples 1-23 of the present application sequentially use kitchen waste liquid culture medium and kitchen waste solid culture medium to ferment and cultivate the Trichoderma harzianum strain, so that the spore production of Trichoderma harzianum reaches 5.5×10 9 cfu / g or more; while the fermentation culture method of Trichoderma provided in Comparative Examples 1-3 was used to ferment the Trichoderma harzianum strain, the spore production of Trichoderma harzianum was only 9.5×10 8 ~2.3×10 9 Therefore, it is shown that the fermentation and cultivation method of Trichoderma provided in the present application not only realizes the effective recycling of kitchen waste, but also realizes the efficient amplification and reproduction of Trichoderma, and has a good application prospect.

[0071] The test results of Examples 1-7 and Comparative Examples 1-3 show that in Example 1-7, the liquid culture medium of kitchen waste prepared by using glucose, ferrous sulfate, magnesium sulfate and kitchen waste liquid was used for liquid fermentation, and the spore production of Trichoderma harzianum obtained was (6.3-6.8)×10 9 cfu / g; Comparative Example 1 used liquid Sabouraud medium for liquid fermentation, and the spore production of Trichoderma harzianum obtained was only 2.3×10 9 cfu / g; Comparative Example 2 used a kitchen waste liquid medium prepared from kitchen waste liquid with an organic matter content of 73 g / L, a nitrogen content of 3.7 g / L, and a phosphorus content of 2.3 g / L for liquid fermentation, and the spore production of the obtained Trichoderma harzianum was only 2.1×10 9 cfu / g; Comparative Example 3 used kitchen waste liquid for liquid fermentation, and the spore production of Trichoderma harzianum was only 9.5×10 8 Therefore, it is explained that the present application uses glucose, ferrous sulfate, magnesium sulfate and kitchen waste liquid to prepare a kitchen waste liquid culture medium, and the dosage of each component and the content of the kitchen waste liquid are controlled within the following ranges: glucose 5-10g / L, ferrous sulfate 0.05-0.1g / L, magnesium sulfate 0.5-1.0g / L; the organic matter content in the kitchen waste liquid is 50-70g / L, the nitrogen content is 4-5g / L, and the phosphorus content is 1-1.5g / L. The prepared kitchen waste liquid culture medium has a good use effect and a significant effect on promoting the growth and reproduction of Trichoderma.

[0072] The test results of Example 2 and Example 8-16 show that the spore production of Trichoderma harzianum obtained in Example 2, Example 8-9, Example 11-12, and Example 14-15 is (6.0-6.8)×10 9 cfu / g; while the spore production of Trichoderma harzianum obtained in Examples 10, 13, and 16 was only (5.5-5.9)×10 9cfu / g, indicating that the present application further controls the addition amount of each component in the kitchen waste solid culture medium within the following range: 60-80 parts of kitchen waste residue phase, 5-15 parts of sawdust and 25-35 parts of rice husk powder, and the obtained kitchen waste solid culture medium has better use effect and higher industrial expansion efficiency of Trichoderma.

[0073] The test results of Example 2 and Examples 17-20 show that with the increase in the amount of kitchen waste liquid culture medium, the spore production of Trichoderma harzianum obtained shows a trend of first increasing and then decreasing; among which the spore production of Trichoderma harzianum obtained in Examples 2 and Examples 18-19 is (6.1-6.8)×10 9 cfu / g; while the spore production of Trichoderma harzianum obtained in Examples 17 and 20 was only (5.6-5.9)×10 9 cfu / g, indicating that the present application further controls the volume ratio of the Trichoderma spore suspension to the kitchen waste liquid culture medium within the range of 1:(50-60), which can improve the fermentation efficiency of Trichoderma and make the reproduction yield of Trichoderma higher.

[0074] The test results of Example 2 and Examples 21-23 show that with the increase in the amount of kitchen waste solid culture medium, the spore production of Trichoderma harzianum obtained shows a trend of first increasing and then decreasing; among which the spore production of Trichoderma harzianum obtained in Examples 2 and Examples 21-22 is (6.1-6.8)×10 9 cfu / g; while the spore production of Trichoderma harzianum obtained in Example 23 was only 5.8×10 9 cfu / g, indicating that the present application further controls the weight ratio of Trichoderma liquid fermentation broth to kitchen waste solid culture medium within the range of 1:(5-10), which can improve the fermentation efficiency of Trichoderma and make the reproduction yield of Trichoderma higher.

[0075] Although the present invention has been described in detail above with general descriptions and specific embodiments, it is obvious to those skilled in the art that some modifications or improvements may be made thereto based on the present invention. Therefore, these modifications or improvements made without departing from the spirit of the present invention all fall within the scope of protection claimed by the present invention.

Claims

1. A kitchen waste liquid culture medium, It is characterized in that The composition includes the following contents: glucose 5-10 g / L, ferrous sulfate 0.05-0.1 g / L, magnesium sulfate 0.5-1.0 g / L, and the remainder is kitchen waste liquid; The kitchen waste liquid has an organic matter content of 50-70 g / L, a nitrogen content of 4-5 g / L, a phosphorus content of 1-1.5 g / L, and a pH value of 4.5-7.

0.

2. A fermentation culture method of Trichoderma, It is characterized in that The kitchen waste liquid culture medium according to claim 1 is adopted.

3. The fermentation and culturing method of Trichoderma according to claim 2, It is characterized in that The following steps are involved: Preparation of Trichoderma spore suspension, liquid fermentation and solid fermentation; Liquid fermentation: inoculating the suspension of Trichoderma spores into the liquid culture medium of kitchen waste, culturing at 25-28° C. and 170-200 rpm in a shaking table for 4-6 days to obtain Trichoderma liquid fermentation liquid; The concentration of Trichoderma spores in the Trichoderma spore suspension is 1×10 8 ~1×10 9 cfu / mL; the volume ratio of the Trichoderma spore suspension to the kitchen waste liquid culture medium is 1: (50-60).

4. The fermentation and culturing method of Trichoderma according to claim 3, It is characterized in that The preparation method of the Trichoderma spore suspension is as follows: inoculating the Trichoderma spores into a PDA culture medium, culturing at 28±2° C. for 4-5 days, and performing light illumination for 8-16 hours and darkness for 8-16 hours alternately during the culturing process; after the culturing is completed, diluting with sterile water to obtain the Trichoderma spore suspension.

5. The fermentation and culturing method of Trichoderma according to claim 3, It is characterized in that The culture medium used in the solid fermentation is a kitchen waste solid culture medium; The kitchen waste solid culture medium comprises the following components in parts by weight: 60-80 parts of kitchen waste residue phase, 0-20 parts of sawdust and 20-40 parts of rice husk powder; The water content of the kitchen waste residue phase is 50-60%, and the pH value is 4.0-6.

5.

6. The fermentation and culturing method of Trichoderma according to claim 5, It is characterized in that The kitchen waste solid culture medium comprises the following components in parts by weight: 60-80 parts of kitchen waste residue phase, 5-15 parts of sawdust and 25-35 parts of rice husk powder.

7. The fermentation and culturing method of Trichoderma according to claim 6, It is characterized in that The method for preparing the kitchen waste solid culture medium comprises: adding other components to the kitchen waste residue phase, sterilizing at 121° C. for 0.5-3 h, and obtaining the kitchen waste solid culture medium.

8. The fermentation and culturing method of Trichoderma according to claim 5, It is characterized in that The specific steps of the solid fermentation are: inoculating the liquid fermentation liquid of Trichoderma into the solid culture medium of kitchen waste, and performing static shallow tray fermentation for 4-7 days at a temperature of 25-30° C. and a humidity of 60-70%, wherein the thickness of the shallow tray fermentation is 2-5 cm, to obtain Trichoderma.

9. The fermentation and culturing method of Trichoderma according to claim 1, It is characterized in that The weight ratio of the Trichoderma liquid fermentation liquid to the kitchen waste solid culture medium is 1: (5-10).

Images