CN111254095B - Compound bacterium liquid, preparation method thereof and application thereof in special fertilizer for corn - Google Patents
Compound bacterium liquid, preparation method thereof and application thereof in special fertilizer for corn Download PDFInfo
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Abstract
The invention relates to a composite bacterial liquid, a preparation method thereof and application thereof in a composite biological bacterial fertilizer special for corn, wherein the composite bacterial liquid is prepared from the following raw materials in parts by weight: 45-55 parts of pleurotus eryngii processing waste extracting solution, 15-20 parts of bacillus mucilaginosus microbial inoculum, 20-25 parts of azotobacter chroococcum microbial inoculum, 5-8 parts of lilac paecilomyces microbial inoculum, 5-10 parts of streptomyces jingyangensis microbial inoculum and 10-15 parts of streptomyces aegiensis Hangzhou variant microbial inoculum; the viable count of the bacillus mucilaginosus agent, the azotobacter chroococcum agent, the lilac paecilomyces agent, the streptomyces jingyangensis agent and the streptomyces griseus Hangzhou variant agent is 2-3 hundred million/mL; according to the invention, the waste pleurotus eryngii processing waste is recycled to prepare the compound bacterium liquid capable of activating nutrients in soil and improving soil fertility.
Description
Technical Field
The invention relates to a composite bacterial liquid, a preparation method thereof and application thereof in a composite biological bacterial fertilizer special for corn.
Background
The pleurotus eryngii has rich nutrition, the content of vegetable protein reaches 25 percent, 18 amino acids are contained, and the polysaccharide has the functions of improving the human immunity and preventing and resisting cancers. Meanwhile, it contains a large amount of oligosaccharide which is 15 times of grifola frondosa, 3.5 times of flammulina velutipes and 2 times of hypsizygus marmoreus, it acts with bifid bacteria in stomach and intestines, it has very good functions of promoting digestion and absorption, pleurotus eryngii is a new rare edible fungus variety which integrates edible, medicinal and dietotherapy, therefore, it is widely planted in China and carries on various processes, such as extraction of polysaccharide and preparation of corresponding food, in the process of pleurotus eryngii, it can produce a large amount of waste materials, such as mushroom feet and broken mushrooms, which contain a large amount of nutrient substances, however, there is no technical means for processing these waste materials, so these waste materials are usually discarded directly, the discarded waste materials not only cause waste of resources, but also will pollute the environment, therefore, it is necessary to develop a technology for processing and reusing these waste materials, to reduce waste of resources.
Corn is an important feed source in the animal husbandry, the aquaculture industry and the like, is one of indispensable raw materials of food, medical health, light industry, chemical industry and the like, has a wide planting range in China, is divided into a plurality of corn areas according to different natural conditions, but is not disturbed by soil fertility reduction and plant diseases and insect pests to a certain extent no matter how the corn is planted, and the phenomena of poor quality and insufficient nutrition are seriously caused by the addition of a large amount of chemical fertilizers, so that the soil is easy to harden due to the use of a large amount of chemical fertilizers, the water and fertilizer retention performance of the soil is also worsened, the normal growth of the corn is seriously influenced, and the economic benefit is poor. In addition, the corn has more than 30 diseases, mainly including large and small spot diseases, head smut, bacterial wilt, virus diseases, stem rot and the like, and the insect pests include corn borer, cutworm, mole cricket, red spider, sorghum striped rice borer, armyworm and the like, and the diseases and the insect pests also greatly influence the yield of the corn.
Disclosure of Invention
The invention aims to provide a composite bacterial liquid, a preparation method thereof and application thereof in a composite biological bacterial fertilizer special for corn. The compound fungus solution can activate nutrients in soil, improve soil fertility, improve disease resistance and insect pest resistance of corn, and is beneficial to yield increase and efficient growth of corn.
The purpose of the invention is realized by the following technical scheme:
the compound bacterium liquid is prepared from the following raw materials in parts by weight: 45-55 parts of pleurotus eryngii processing waste extracting solution, 15-20 parts of bacillus mucilaginosus microbial inoculum, 20-25 parts of azotobacter chroococcum microbial inoculum, 5-8 parts of lilac paecilomyces microbial inoculum, 5-10 parts of streptomyces jingyangensis microbial inoculum and 10-15 parts of streptomyces aegiensis Hangzhou variant microbial inoculum; the viable count of the bacillus mucilaginosus agent, the azotobacter chroococcum agent, the lilac paecilomyces agent, the streptomyces jingyangensis agent and the streptomyces griseus Hangzhou variant agent is 2-3 hundred million/mL.
The preparation method of the compound bacterium liquid comprises the following steps:
(1) preparing pleurotus eryngii processing waste extracting solution; (2) activating each strain; (3) preparing Bacillus mucilaginosus seed suspension, azotobacter chroococcum seed suspension, lilac paecilomyces seed suspension, streptomyces jingyangensis seed suspension, and streptomyces griseus Hangzhou variety seed suspension; (4) preparing Bacillus mucilaginosus agent, azotobacter nodosum agent, lilac paecilomyces agent, streptomyces jingyangensis agent and streptomyces griseus Hangzhou variant agent; (5) preparing a bacterial liquid;
wherein the method for preparing the bacillus mucilaginosus microbial inoculum, the azotobacter chroococcum microbial inoculum, the lilac paecilomyces microbial inoculum, the Jingyang streptomyces microbial inoculum and the light gray streptomyces Hangzhou variant microbial inoculum in the step (4) comprises the following steps:
taking part of the pleurotus eryngii processing waste extracting solution obtained in the step (1), adjusting the pH value of the extracting solution to 7.2-7.4, and performing moist heat sterilization at the temperature of 121-;
taking 5 triangular flasks with the volume of 250mL, and placing the obtained culture solution into the triangular flasks, wherein the volume of the culture solution in each triangular flask is 80-90 mL; then, respectively inoculating the bacillus mucilaginosus seed suspension, the azotobacter chroococcum seed suspension, the lilac paecilomyces seed suspension, the Jingyang streptomyces seed suspension and the light gray streptomyces Hangzhou variety seed suspension obtained in the step (3) into corresponding triangular flasks, wherein the inoculation amount in each triangular flask is 4-6%; after inoculation, carrying out shaking culture at the conditions of 28-30 ℃ and the rotation speed of 100-250rpm, wherein bacillus mucilaginosus and azotobacter chroococcum are cultured for 48-72h, and paecilomyces lilacinus, streptomyces jingyangensis and streptomyces shallowlii are cultured for 7-9 days, so that the viable count of each microbial inoculum is 2-3 hundred million/mL, and thus the bacillus mucilaginosus microbial inoculum, the azotobacter chroococcum microbial inoculum, the paecilomyces lilacinus microbial inoculum, the streptomyces jingyangensis microbial inoculum and the streptomyces shallowlii Hangzhou microbial inoculum are respectively obtained;
the method for preparing the bacterial liquid in the step (5) comprises the following steps: taking part of the pleurotus eryngii processing waste extracting solution obtained in the step (1), and carrying out damp-heat sterilization on the extracting solution at the temperature of 121-; uniformly mixing the sterilized pleurotus eryngii processing waste extracting solution, a bacillus mucilaginosus microbial inoculum, a brown nodule azotobacter microbial inoculum, a lilac paecilomyces microbial inoculum, a Jingyang streptomyces microbial inoculum and a Streptomyces griseus Hangzhou variant microbial inoculum according to parts by weight to obtain the compound bacterial liquid.
The composite bacterial liquid is applied to the preparation of the composite biological bacterial fertilizer special for corn.
The composite biological bacterial fertilizer special for the corn comprises the following components in parts by weight:
30-40 parts of composite bacteria liquid, 15-20 parts of bran, 15-20 parts of attapulgite, 30-40 parts of peat, 20-25 parts of bentonite and 15-25 parts of bean cakes.
The preparation method of the composite biological bacterial fertilizer special for the corns comprises the following steps:
i, preparing the compound bacterium liquid;
and II, crushing bran, attapulgite, bentonite and bean cakes, mixing with peat and the prepared composite bacterial liquid under stirring, and drying at 25-30 ℃ until the water content is 5-8% to obtain the composite biological bacterial fertilizer special for the corn.
Compared with the prior art, the invention has the advantages that:
1. according to the invention, the waste pleurotus eryngii processing waste is recycled to prepare the compound bacterium liquid capable of activating nutrients in soil and improving soil fertility.
2. The composite bacterial liquid and the composite biological bacterial fertilizer special for the corn can improve the fertility of soil, fully decompose indissolvable phosphorus, potassium, mineral substances and the like in the soil, promote the corn plants to absorb nutrient components in the soil, promote the growth of the corn plants, reduce the use of chemical fertilizers and improve the quality of the corn.
3. The compound bacteria liquid and the compound biological bacterial fertilizer special for corn can also inhibit the growth and propagation of harmful microorganisms, achieve the effects of restoring soil and preventing soil hardening, and enable the soil environment to be in a loose nutritional state.
4. The compound bacterial liquid and the compound biological bacterial fertilizer special for the corn can also protect the root system of the corn from being invaded by harmful microorganisms, and improve the disease resistance and insect resistance of the corn, such as the capability of the corn for resisting large and small spots, bacterial wilt and stem rot.
5. The pleurotus eryngii processing waste extracting solution disclosed by the invention contains a large amount of amino acids, carbohydrates and trace elements, has a synergistic promotion effect by combining with various bacteria, and can promote corn plants to absorb and utilize elements such as nitrogen, phosphorus and potassium.
6. The composite biological bacterial fertilizer special for the corn can also improve the drought resistance of corn plants and improve the water retention performance of soil.
Detailed Description
The present invention will be described in detail with reference to the following examples:
the compound bacterium liquid is prepared from the following raw materials in parts by weight: 45-55 parts of pleurotus eryngii processing waste extracting solution, 15-20 parts of bacillus mucilaginosus microbial inoculum, 20-25 parts of azotobacter chroococcum microbial inoculum, 5-8 parts of lilac paecilomyces microbial inoculum, 5-10 parts of streptomyces jingyangensis microbial inoculum and 10-15 parts of streptomyces aegiensis Hangzhou variant microbial inoculum; the viable count of the bacillus mucilaginosus agent, the azotobacter chroococcum agent, the lilac paecilomyces agent, the streptomyces jingyangensis agent and the streptomyces griseus Hangzhou variant agent is 2-3 hundred million/mL.
The preparation method of the compound bacterium liquid comprises the following steps:
(1) preparing pleurotus eryngii processing waste extracting solution; (2) activating each strain; (3) preparing Bacillus mucilaginosus seed suspension, azotobacter chroococcum seed suspension, lilac paecilomyces seed suspension, streptomyces jingyangensis seed suspension, and streptomyces griseus Hangzhou variety seed suspension; (4) preparing Bacillus mucilaginosus agent, azotobacter nodosum agent, lilac paecilomyces agent, streptomyces jingyangensis agent and streptomyces griseus Hangzhou variant agent; (5) preparing a bacterial liquid;
wherein the method for preparing the pleurotus eryngii processing waste extracting solution in the step (1) comprises the following steps:
cleaning mushroom feet and crushed mushrooms generated in the pleurotus eryngii processing process, crushing the mushroom feet and the crushed mushrooms into 40-50 meshes, adding water according to the weight ratio of 2:3 of the pleurotus eryngii to the water, heating the mixture to 90-98 ℃, decocting the mixture for 2-3 hours at the temperature, cooling and filtering the mixture, and collecting filtrate; centrifuging the collected filtrate at the rotating speed of 1500-; concentrating the obtained supernatant by falling film evaporation with a falling film evaporator until the refractive index of the solution reaches 35-45% to obtain a concentrated solution; then, adding 80-90% of ethanol into the concentrated solution, and modifying the concentrated solution, wherein the volume ratio of the added ethanol to the concentrated solution is 3: 2; filtering the denatured concentrated solution to remove precipitate generated after denaturation, and collecting filtrate; performing rotary evaporation on the filtrate to remove ethanol; and (3) performing tangential flow filtration on the solution obtained after rotary evaporation by using a filter membrane with the molecular weight cutoff of 5-10K, and collecting filtrate to obtain the pleurotus eryngii processing waste extracting solution.
The method for activating each strain in the step (2) comprises the following steps:
respectively inoculating bacillus mucilaginosus and azotobacter chroococcum on a slant containing a solid culture medium A, and separately culturing for 48-72h, wherein the culture temperature is controlled at 25-35 ℃;
wherein the solid culture medium A is K2HPO4、KH2PO4、MgSO4·7H2O、CaCO3·2H2O、Na2Mo·2H2O, yeast extract, mannitol, FeCl3And agar to prepare a culture medium;
the preparation method of the solid culture medium A comprises the following steps: will K2HPO4 0.75-0.85g/L；KH2PO4 0.15-0.25g/L；MgSO4·7H2O 0.15-0.25g/L；CaCO3·2H2O 0.05-0.15g/L；Na2Mo·2H20.01-0.02g/L of O, 0.45-0.55g/L of yeast extract; 15-25g/L of mannitol; FeCl30.01-0.02g/L agar 10-20g/L, and distilled water in balance, adjusting pH to 7.0-7.2, and sterilizing at 115-122 deg.C for 15-20 min.
Inoculating Paecilomyces lilacinus to slant culture medium containing PDA slant culture medium, and culturing at 25-28 deg.C for 5-7 days;
the preparation method of the PDA slant culture medium comprises the following steps: mixing 20% potato juice 1L, glucose 20g, KH2PO4 3g、MgSO4·7H2O1.5 g, Vitanib B1 (thiamine) Trace 8mg, and agar 20g were mixed, pH was adjusted to natural pH, and autoclaved at 121 ℃ for 15 min.
Inoculating Streptomyces jingyangensis on a slant containing Gaoshi synthetic first agar medium, and culturing for 5-7 days at 28-30 deg.C;
the Hangzhou variety of Streptomyces lividans is inoculated on a slant containing Gao's synthetic first agar medium and cultured for 5-7 days at a temperature controlled at 30 ℃.
The preparation method of the Gao's synthetic agar medium I comprises the following steps: mixing KNO31g of soluble starch, 20g of soluble starch and K2HPO40.5g、MgSO4·7H2O 0.5g、NaCl 0.5g、FeSO40.01g of agar, 20g of agar and 1L of distilled water are mixed, the pH value is adjusted to 7.2-7.4, and then the mixture is sterilized for 15-20min at the temperature of 115 ℃ and 122 ℃.
The bacillus mucilaginosus used in the invention has a preservation number of GIM1.16, the azotobacter chroococcum has a preservation number of GIM1.272, the paecilomyces lilacinus has a preservation number of GIM3.405, the streptomyces jingyangensis has a preservation number of GIM4.92, and the streptomyces griseus Hangzhou variant has a preservation number of GIM4.96, and the bacillus mucilaginosus (GIM1.16), the azotobacter chroococcum (GIM1.272), the paecilomyces lilacinus (GIM3.405), the streptomyces jingyangensis (GIM4.92) and the streptomyces griseus Hangzhou variant (GIM4.96) are all purchased from Guangdong province microorganism culture collection center.
The method for preparing the bacillus mucilaginosus seed suspension, the azotobacter chroococcum seed suspension, the lilac paecilomyces seed suspension, the Jingyang streptomyces seed suspension and the light gray streptomyces Hangzhou variant seed suspension comprises the following steps:
selecting 1-ring activated Bacillus mucilaginosus, azotobacter chroococcum, paecilomyces lilacinus, streptomyces jingyangensis and streptomyces griseus Hangzhou varieties in the step (2) by using inoculating rings, and correspondingly inoculating the strains into 5 triangular flasks filled with 100-150mL liquid culture medium A, liquid culture medium A, PDA, Gauss synthetic first liquid culture medium and Gauss synthetic first liquid culture medium for culturing, wherein the Bacillus mucilaginosus and the azotobacter chroococcum are subjected to shaking culture for 24-48h under the conditions of 200rpm of 150-charge and 25-35 ℃; culturing the lilac paecilomyces on a shaking table for 5-7 days at the temperature of 25-28 ℃ and 200rpm of 150-; shaking culturing Streptomyces jingyangensis for 5-7 days at 150-; shaking and culturing Hangzhou variety of Streptomyces lividans in a shaker at 150-; wherein the liquid culture medium A is K2HPO4、KH2PO4、MgSO4·7H2O、CaCO3·2H2O、Na2Mo·2H2O, yeast extract, mannitol, FeCl3Mixing to obtain the culture medium.
The preparation method of the liquid culture medium A comprises the following steps: will K2HPO4 0.75-0.85g/L；KH2PO4 0.15-0.25g/L；MgSO4·7H2O 0.15-0.25g/L；CaCO3·2H2O 0.05-0.15g/L；Na2Mo·2H20.01-0.02g/L of O, 0.45-0.55g/L of yeast extract; 15-25g/L of mannitol; FeCl30.01-0.02 g/L; mixing the rest of distilled water, adjusting pH to 7.0-7.2, and sterilizing at 115-122 deg.C for 15-20 min.
The preparation method of the PDA liquid culture medium comprises the following steps: 1L of 20 percent potato juice, 20g of glucose,KH2PO4 3g、MgSO4·7H2O1.5 g and Vitanib B1 (thiamine) Trace 8mg, mixed at natural pH, autoclaved at 121 ℃ for 15 min.
The preparation method of the Gao's synthetic No. I liquid culture medium comprises the following steps: mixing KNO31g of soluble starch, 20g of soluble starch and K2HPO40.5g、MgSO4·7H2O 0.5g、NaCl 0.5g、FeSO40.01g and 1L of distilled water, adjusting the pH value to 7.2-7.4, and then sterilizing at 115-122 ℃ for 15-20 min.
The method for preparing the bacillus mucilaginosus agent, the brown nodule azotobacter agent, the lilac paecilomyces agent, the Jingyang streptomyces agent and the light gray streptomyces Hangzhou variant agent comprises the following steps:
taking part of the pleurotus eryngii processing waste extracting solution obtained in the step (1), adjusting the pH value of the extracting solution to 7.2-7.4, and performing moist heat sterilization at the temperature of 121-;
taking 5 triangular flasks with the volume of 250mL, and placing the obtained culture solution into the triangular flasks, wherein the volume of the culture solution in each triangular flask is 80-90 mL; then, respectively inoculating the bacillus mucilaginosus seed suspension, the azotobacter chroococcum seed suspension, the lilac paecilomyces seed suspension, the Jingyang streptomyces seed suspension and the light gray streptomyces Hangzhou variety seed suspension obtained in the step (3) into corresponding triangular flasks, wherein the inoculation amount in each triangular flask is 4-6%; after inoculation, carrying out shaking culture at the conditions of 28-30 ℃ and the rotation speed of 100-250rpm, wherein bacillus mucilaginosus and azotobacter chroococcum are cultured for 48-72h, and paecilomyces lilacinus, streptomyces jingyangensis and streptomyces shallowlii are cultured for 7-9 days, so that the viable count of each microbial inoculum is 2-3 hundred million/mL, and thus the bacillus mucilaginosus microbial inoculum, the azotobacter chroococcum microbial inoculum, the paecilomyces lilacinus microbial inoculum, the streptomyces jingyangensis microbial inoculum and the streptomyces shallowlii Hangzhou microbial inoculum are respectively obtained;
the method for preparing the bacterial liquid in the step (5) comprises the following steps: taking part of the pleurotus eryngii processing waste extracting solution obtained in the step (1), and carrying out damp-heat sterilization on the extracting solution at the temperature of 121-; uniformly mixing the sterilized pleurotus eryngii processing waste extracting solution, a bacillus mucilaginosus microbial inoculum, a brown nodule azotobacter microbial inoculum, a lilac paecilomyces microbial inoculum, a Jingyang streptomyces microbial inoculum and a Streptomyces griseus Hangzhou variant microbial inoculum according to parts by weight to obtain the compound bacterial liquid.
The composite bacterial liquid can be directly applied to the root of a corn plant after being diluted by water, and the volume ratio of the composite bacterial liquid to the added water is 1:20 during dilution.
The composite bacterial liquid can also be mixed with an adsorbent to prepare a solid bacterial fertilizer. The composite bacterial liquid can be used for preparing the composite biological bacterial fertilizer special for corn. The composite biological bacterial fertilizer special for the corn comprises the following components in parts by weight: 30-40 parts of composite bacteria liquid, 15-20 parts of bran, 15-20 parts of attapulgite, 30-40 parts of peat, 20-25 parts of bentonite and 15-25 parts of bean cakes.
The preparation method of the composite biological bacterial fertilizer special for the corn comprises the following steps:
i, preparing the compound bacterium liquid;
and II, crushing bran, attapulgite, bentonite and bean cakes, mixing with peat and the prepared composite bacterial liquid under stirring, and drying at 25-30 ℃ until the water content is 5-8% to obtain the composite biological bacterial fertilizer special for the corn.
The invention is explained in more detail below with reference to specific examples:
the Bacillus mucilaginosus used in the following examples of the present invention has a accession number of GIM1.16, the Azotobacter sphaeroides has a accession number of GIM1.272, the Paecilomyces lilacinus has a accession number of GIM3.405, the Streptomyces jingyangensis has a accession number of GIM4.92, and the Streptomyces lividans Hangzhou variety has a accession number of GIM4.96, and the Bacillus mucilaginosus (GIM1.16), the Azotobacter sphaeroides (GIM1.272), the Paecilomyces lilacinus (GIM3.405), the Streptomyces jingyangensis (GIM4.92), and the Streptomyces lividans Hangzhou variety (GIM4.96) were all purchased from Guangdong province microorganism culture collection.
The following examples of the present invention use media: a solid culture medium A, PDA slant culture medium, a Gao's synthetic No. I agar culture medium, a liquid culture medium A, PDA liquid culture medium and a Gao's synthetic No. I liquid culture medium;
wherein the solid culture medium A is K2HPO4、KH2PO4、MgSO4·7H2O、CaCO3·2H2O、Na2Mo·2H2O, yeast extract, mannitol, FeCl3And agar to obtain culture medium; the preparation method of the solid culture medium A comprises the following steps: will K2HPO4 0.75-0.85g/L；KH2PO4 0.15-0.25g/L；MgSO4·7H2O 0.15-0.25g/L；CaCO3·2H2O 0.05-0.15g/L；Na2Mo·2H20.01-0.02g/L of O, 0.45-0.55g/L of yeast extract; 15-25g/L of mannitol; FeCl30.01-0.02g/L agar 10-20g/L, and distilled water in balance, adjusting pH to 7.0-7.2, and sterilizing at 115-122 deg.C for 15-20 min.
The preparation method of the PDA slant culture medium comprises the following steps: mixing 20% potato juice 1L, glucose 20g, KH2PO4 3g、MgSO4·7H2O1.5 g, Vitanib B1 (thiamine) Trace 8mg, and agar 20g were mixed, pH was adjusted to natural pH, and autoclaved at 121 ℃ for 15 min.
The preparation method of the Gao's synthetic agar medium I comprises the following steps: mixing KNO31g of soluble starch, 20g of soluble starch and K2HPO40.5g、MgSO4·7H2O 0.5g、NaCl 0.5g、FeSO40.01g of agar, 20g of agar and 1L of distilled water are mixed, the pH value is adjusted to 7.2-7.4, and then the mixture is sterilized for 15-20min at the temperature of 115 ℃ and 122 ℃.
The liquid culture medium A is K2HPO4、KH2PO4、MgSO4·7H2O、CaCO3·2H2O、Na2Mo·2H2O, yeast extract, mannitol, FeCl3Mixing to obtain the culture medium. The preparation method of the liquid culture medium A comprises the following steps: will K2HPO4 0.75-0.85g/L；KH2PO4 0.15-0.25g/L；MgSO4·7H2O 0.15-0.25g/L；CaCO3·2H2O 0.05-0.15g/L；Na2Mo·2H20.01-0.02g/L of O, 0.45-0.55g/L of yeast extract; 15-25g/L of mannitol; FeCl30.01-0.02 g/L; the balance of distilled waterMixing, adjusting pH to 7.0-7.2, and sterilizing at 115-122 deg.C for 15-20 min.
The preparation method of the PDA liquid culture medium comprises the following steps: mixing 20% potato juice 1L, glucose 20g, KH2PO4 3g、MgSO4·7H2O1.5 g and Vitanib B1 (thiamine) Trace 8mg, mixed at natural pH, autoclaved at 121 ℃ for 15 min.
The preparation method of the Gao's synthetic No. I liquid culture medium comprises the following steps: mixing KNO31g of soluble starch, 20g of soluble starch and K2HPO40.5g、MgSO4·7H2O 0.5g、NaCl 0.5g、FeSO40.01g and 1L of distilled water, adjusting the pH value to 7.2-7.4, and then sterilizing at 115-122 ℃ for 15-20 min.
The first embodiment is as follows:
the compound bacterium liquid is prepared from the following raw materials in parts by weight: 45 parts of pleurotus eryngii processing waste extracting solution, 20 parts of bacillus mucilaginosus microbial inoculum, 20 parts of azotobacter chroococcum microbial inoculum, 8 parts of lilac paecilomyces microbial inoculum, 5 parts of streptomyces jingyangensis microbial inoculum and 15 parts of streptomyces griseus Hangzhou variant microbial inoculum; the viable count of the bacillus mucilaginosus agent, the azotobacter chroococcum agent, the lilac paecilomyces agent, the streptomyces jingyangensis agent and the streptomyces griseus Hangzhou variant agent is 2-3 hundred million/mL.
The preparation method of the compound bacterium liquid comprises the following steps:
(1) preparing pleurotus eryngii processing waste extracting solution; (2) activating each strain; (3) preparing Bacillus mucilaginosus seed suspension, azotobacter chroococcum seed suspension, lilac paecilomyces seed suspension, streptomyces jingyangensis seed suspension, and streptomyces griseus Hangzhou variety seed suspension; (4) preparing Bacillus mucilaginosus agent, azotobacter nodosum agent, lilac paecilomyces agent, streptomyces jingyangensis agent and streptomyces griseus Hangzhou variant agent; (5) preparing a bacterial liquid;
wherein the method for preparing the pleurotus eryngii processing waste extracting solution in the step (1) comprises the following steps:
cleaning mushroom feet and crushed mushrooms generated in the pleurotus eryngii processing process, crushing the mushroom feet and the crushed mushrooms into 40-50 meshes, adding water according to the weight ratio of 2:3 of the pleurotus eryngii to the water, heating the mixture to 90 ℃, decocting the mixture for 3 hours at the temperature, cooling and filtering the mixture, and collecting filtrate; centrifuging the collected filtrate at 1500r/min for 15min, and collecting supernatant; the obtained supernatant is evaporated and concentrated by falling film evaporation by a falling film evaporator until the refractive index of the solution reaches 35 percent to obtain concentrated solution; then, adding 90% ethanol into the concentrated solution, and modifying the concentrated solution, wherein the volume ratio of the added ethanol to the concentrated solution is 3: 2; filtering the denatured concentrated solution to remove precipitate generated after denaturation, and collecting filtrate; performing rotary evaporation on the filtrate to remove ethanol; and (3) performing tangential flow filtration on the solution obtained after rotary evaporation by using a filter membrane with the molecular weight cutoff of 5K, and collecting filtrate to obtain the pleurotus eryngii processing waste extracting solution.
The method for activating each strain in the step (2) comprises the following steps:
respectively inoculating bacillus mucilaginosus and azotobacter chroococcum on a slant containing a solid culture medium A, and separately culturing for 48h, wherein the culture temperature is controlled at 35 ℃ during culture;
inoculating paecilomyces lilacinus on a slant containing PDA slant culture medium, and culturing for 5 days at 28 deg.C;
inoculating streptomyces jingyangensis on a slant containing Gao's synthetic first agar medium, and culturing for 5 days at the culture temperature of 30 ℃;
the Hangzhou variety of Streptomyces lividans is inoculated on a slant containing a Gao's synthetic first agar medium and cultured for 5 days at a temperature controlled at 30 ℃.
The method for preparing the bacillus mucilaginosus seed suspension, the azotobacter chroococcum seed suspension, the lilac paecilomyces seed suspension, the Jingyang streptomycete seed suspension and the light gray streptomycete Hangzhou variety seed suspension comprises the following steps:
selecting 1 ring of activated bacillus mucilaginosus, azotobacter chroococcum, paecilomyces lilacinus, streptomyces jingyangensis and streptomyces griseus Hangzhou varieties in the step (2) by using inoculating rings, and correspondingly inoculating the bacillus mucilaginosus, azotobacter chroococcum, paecilomyces lilacinus, streptomyces jingyangensis and streptomyces griseus into 5 triangular flasks filled with 100mL of liquid culture medium A, A, PDA of liquid culture medium, I-Gao-synthetic liquid culture medium and I-Gao-synthetic liquid culture medium respectively for culture, wherein the bacillus mucilaginosus and the azotobacter chroococcum are subjected to shaking culture for 24 hours at the conditions of 200rpm and 35 ℃; culturing the lilac paecilomyces for 5 days at the conditions of 200rpm and 28 ℃ by shaking on a shaking table; shaking culturing Streptomyces jingyangensis for 5 days at 200rpm and 30 ℃; shaking and culturing the Hangzhou variety of the streptomyces aegaensis for 5 days at the temperature of 30 ℃ and at the rpm of 200, thus respectively obtaining a bacillus mucilaginosus seed suspension, a azotobacter chroococcum seed suspension, a paecilomyces lilacinus seed suspension, a streptomyces jingyangensis seed suspension and a streptomyces aegaensis Hangzhou variety seed suspension;
the method for preparing the bacillus mucilaginosus agent, the brown nodule azotobacter agent, the lilac paecilomyces agent, the Jingyang streptomyces agent and the light gray streptomyces Hangzhou variant agent comprises the following steps:
taking part of the pleurotus eryngii processing waste extracting solution obtained in the step (1), adjusting the pH value of the extracting solution to 7.2, and performing damp-heat sterilization at 121 ℃ for 20min to obtain a culture solution;
taking 5 triangular flasks with the volume of 250mL, and placing the obtained culture solution into the triangular flasks, wherein the volume of the culture solution in each triangular flask is 80 mL; then, respectively inoculating the bacillus mucilaginosus seed suspension, the azotobacter chroococcum seed suspension, the lilac paecilomyces seed suspension, the Jingyang streptomyces seed suspension and the light gray streptomyces Hangzhou variety seed suspension obtained in the step (3) into corresponding triangular flasks, wherein the inoculation amount in each triangular flask is 4%; after inoculation, carrying out shaking culture at the temperature of 30 ℃ and the rotating speed of 250rpm, wherein bacillus mucilaginosus and azotobacter chroococcum are cultured for 48h, and paecilomyces lilacinus, streptomyces jingyangensis and streptomyces griseus are cultured for 7 days, so that the viable count of each microbial inoculum is 2-3 hundred million/mL, and thus the bacillus mucilaginosus microbial inoculum, the azotobacter chroococcum microbial inoculum, the paecilomyces lilacinus microbial inoculum, the streptomyces jingyangensis microbial inoculum and the streptomyces griseus Hangzhou microbial inoculum are respectively obtained;
the method for preparing the bacterial liquid in the step (5) comprises the following steps: taking part of the pleurotus eryngii processing waste extracting solution obtained in the step (1), and carrying out damp-heat sterilization on the extracting solution for 20min at the temperature of 121 ℃; uniformly mixing the sterilized pleurotus eryngii processing waste extracting solution, a bacillus mucilaginosus microbial inoculum, a brown nodule azotobacter microbial inoculum, a lilac paecilomyces microbial inoculum, a Jingyang streptomyces microbial inoculum and a Streptomyces griseus Hangzhou variant microbial inoculum according to parts by weight to obtain the compound bacterial liquid.
Example two:
the compound bacterium liquid is prepared from the following raw materials in parts by weight: 50 parts of pleurotus eryngii processing waste extracting solution, 17 parts of bacillus mucilaginosus agent, 22 parts of azotobacter chroococcum agent, 6 parts of lilac paecilomyces variotii agent, 8 parts of streptomyces jingyangensis agent and 12 parts of streptomyces griseus Hangzhou variant agent; the viable count of the Bacillus mucilaginosus agent, the azotobacter nodosum agent, the paecilomyces lilacinus agent, the streptomyces jingyangensis agent and the streptomyces griseus Hangzhou variant agent is 2-3 hundred million/mL.
The preparation method of the compound bacterium liquid comprises the following steps:
(1) preparing pleurotus eryngii processing waste extracting solution; (2) activating each strain; (3) preparing Bacillus mucilaginosus seed suspension, azotobacter chroococcum seed suspension, lilac paecilomyces seed suspension, streptomyces jingyangensis seed suspension, and streptomyces griseus Hangzhou variety seed suspension; (4) preparing Bacillus mucilaginosus agent, azotobacter nodosum agent, lilac paecilomyces agent, streptomyces jingyangensis agent and streptomyces griseus Hangzhou variant agent; (5) preparing a bacterial liquid;
wherein the method for preparing the pleurotus eryngii processing waste extracting solution in the step (1) comprises the following steps:
cleaning mushroom feet and crushed mushrooms generated in the pleurotus eryngii processing process, crushing the mushroom feet and the crushed mushrooms into 40-50 meshes, adding water according to the weight ratio of the pleurotus eryngii to the water of 2:3, heating the mixture to 95 ℃, decocting the mixture for 2.5 hours at the temperature, cooling and filtering the mixture, and collecting filtrate; centrifuging the collected filtrate at 2000r/min for 12min, and collecting supernatant; the obtained supernatant is evaporated and concentrated by falling film evaporation by a falling film evaporator until the refractive index of the solution reaches 40 percent to obtain concentrated solution; then, 85% of ethanol is added into the concentrated solution to denature the concentrated solution, wherein the volume ratio of the added ethanol to the concentrated solution is 3: 2; filtering the denatured concentrated solution to remove precipitate generated after denaturation, and collecting filtrate; performing rotary evaporation on the filtrate to remove ethanol; and (3) performing tangential flow filtration on the solution obtained after rotary evaporation by using a filter membrane with the molecular weight cutoff of 10K, and collecting filtrate to obtain the pleurotus eryngii processing waste extracting solution.
The method for activating each strain in the step (2) comprises the following steps:
respectively inoculating bacillus mucilaginosus and azotobacter chroococcum on a slant containing a solid culture medium A, and separately culturing for 60h, wherein the culture temperature is controlled at 28 ℃ during culture;
inoculating paecilomyces lilacinus on a slant containing PDA slant culture medium, and culturing for 6 days at 27 deg.C;
inoculating streptomyces jingyangensis on a slant containing a Gao's synthetic first agar medium to culture for 6 days, wherein the culture temperature is controlled at 29 ℃;
the Hangzhou variety of Streptomyces lividans is inoculated on a slant containing a Gao's synthetic first agar medium and cultured for 6 days at a temperature controlled at 30 ℃.
The method for preparing the bacillus mucilaginosus seed suspension, the azotobacter chroococcum seed suspension, the lilac paecilomyces seed suspension, the Jingyang streptomyces seed suspension and the light gray streptomyces Hangzhou variant seed suspension comprises the following steps:
respectively selecting 1 ring of activated bacillus mucilaginosus, azotobacter chroococcum, paecilomyces lilacinus, streptomyces jingyangensis and streptomyces griseus Hangzhou varieties in the step (2) by using inoculating rings, and correspondingly inoculating the varieties into 5 triangular flasks filled with 120mL of liquid culture medium A, A, PDA of liquid culture medium, Gao's synthetic first liquid culture medium and Gao's synthetic first liquid culture medium for culture, wherein the bacillus mucilaginosus and the azotobacter chroococcum are subjected to shaking culture for 36 hours at 180rpm and 28 ℃; shaking culturing Paecilomyces lilacinus at 180rpm and 27 deg.C for 6 days; shaking culturing Streptomyces jingyangensis on a shaker for 6 days at 180rpm and 29 ℃; shaking and culturing the Hangzhou variety of the streptomyces aegaensis for 6 days at 180rpm and 30 ℃ in a shaking way to respectively obtain a bacillus mucilaginosus seed suspension, a azotobacter chroococcum seed suspension, a paecilomyces lilacinus seed suspension, a streptomyces jingyangensis seed suspension and a streptomyces aegaensis Hangzhou variety seed suspension;
the method for preparing the bacillus mucilaginosus agent, the brown nodule azotobacter agent, the lilac paecilomyces agent, the Jingyang streptomyces agent and the light gray streptomyces Hangzhou variant agent comprises the following steps:
taking part of the pleurotus eryngii processing waste extracting solution obtained in the step (1), adjusting the pH value of the extracting solution to 7.3, and performing damp-heat sterilization at 122 ℃ for 15min to obtain a culture solution;
taking 5 triangular flasks of 250mL, and placing the obtained culture solution into the triangular flasks, wherein the volume of the culture solution in each triangular flask is 85 mL; then, respectively inoculating the bacillus mucilaginosus seed suspension, the azotobacter chroococcum seed suspension, the lilac paecilomyces seed suspension, the Jingyang streptomyces seed suspension and the light gray streptomyces Hangzhou variety seed suspension obtained in the step (3) into corresponding triangular flasks, wherein the inoculation amount in each triangular flask is 5%; after inoculation, carrying out shaking culture at the temperature of 28 ℃ and the rotating speed of 200rpm, wherein bacillus mucilaginosus and azotobacter chroococcum are cultured for 60h, and paecilomyces lilacinus, streptomyces jingyangensis and streptomyces griseus Hangzhou varieties are cultured for 8 days, so that the number of live bacteria of each microbial inoculum is 2-3 hundred million/mL, and thus the bacillus mucilaginosus microbial inoculum, the azotobacter chroococcum microbial inoculum, the paecilomyces lilacinus microbial inoculum, the streptomyces jingyangensis microbial inoculum and the streptomyces griseus Hangzhou variety microbial inoculum are obtained respectively;
the method for preparing the bacterial liquid in the step (5) comprises the following steps: taking part of the pleurotus eryngii processing waste extracting solution obtained in the step (1), and carrying out damp-heat sterilization on the extracting solution for 15min at 122 ℃; uniformly mixing the sterilized pleurotus eryngii processing waste extracting solution, a bacillus mucilaginosus microbial inoculum, a brown nodule azotobacter microbial inoculum, a lilac paecilomyces microbial inoculum, a Jingyang streptomyces microbial inoculum and a Streptomyces griseus Hangzhou variant microbial inoculum according to parts by weight to obtain the compound bacterial liquid.
Example three:
the compound bacterium liquid is prepared from the following raw materials in parts by weight: 55 parts of pleurotus eryngii processing waste extracting solution, 15 parts of bacillus mucilaginosus fungicide, 25 parts of azotobacter chroococcum fungicide, 5 parts of lilac paecilomyces fungicide, 10 parts of streptomyces jingyangensis fungicide and 10 parts of streptomyces aegiensis Hangzhou variant fungicide; the viable count of the bacillus mucilaginosus agent, the azotobacter chroococcum agent, the lilac paecilomyces agent, the streptomyces jingyangensis agent and the streptomyces griseus Hangzhou variant agent is 2-3 hundred million/mL.
The preparation method of the compound bacterium liquid comprises the following steps:
(1) preparing pleurotus eryngii processing waste extracting solution; (2) activating each strain; (3) preparing Bacillus mucilaginosus seed suspension, azotobacter chroococcum seed suspension, lilac paecilomyces seed suspension, streptomyces jingyangensis seed suspension, and streptomyces griseus Hangzhou variety seed suspension; (4) preparing Bacillus mucilaginosus agent, azotobacter nodosum agent, lilac paecilomyces agent, streptomyces jingyangensis agent and streptomyces griseus Hangzhou variant agent; (5) preparing a bacterial liquid;
wherein the method for preparing the pleurotus eryngii processing waste extracting solution in the step (1) comprises the following steps:
cleaning mushroom feet and crushed mushrooms generated in the pleurotus eryngii processing process, crushing the mushroom feet and the crushed mushrooms into 40-50 meshes, adding water according to the weight ratio of 2:3 of the pleurotus eryngii to the water, heating the mixture to 98 ℃, decocting the mixture for 2 hours at the temperature, cooling and filtering the mixture, and collecting filtrate; centrifuging the collected filtrate at 2500r/min for 10min, and collecting supernatant; the obtained supernatant is evaporated and concentrated by falling film evaporation of a falling film evaporator until the refractive index of the solution reaches 45 percent to obtain concentrated solution; then, adding 80% ethanol into the concentrated solution, and modifying the concentrated solution, wherein the volume ratio of the added ethanol to the concentrated solution is 3: 2; filtering the denatured concentrated solution to remove precipitate generated after denaturation, and collecting filtrate; performing rotary evaporation on the filtrate to remove ethanol; and (3) performing tangential flow filtration on the solution obtained after rotary evaporation by using a filter membrane with the molecular weight cutoff of 10K, and collecting filtrate to obtain the pleurotus eryngii processing waste extracting solution.
The method for activating each strain in the step (2) comprises the following steps:
respectively inoculating bacillus mucilaginosus and azotobacter chroococcum on a slant containing a solid culture medium A, and separately culturing for 72h, wherein the culture temperature is controlled at 25 ℃ during culture;
inoculating paecilomyces lilacinus on a slant containing a PDA slant culture medium, and culturing for 7 days at the culture temperature of 25 ℃;
inoculating streptomyces jingyangensis on a slant containing a Gao's synthetic first agar medium to culture for 7 days, wherein the culture temperature is controlled at 28 ℃;
the Hangzhou variety of Streptomyces lividans is inoculated on a slant containing a Gao's synthetic first agar medium and cultured for 7 days at a temperature controlled at 30 ℃.
The method for preparing the bacillus mucilaginosus seed suspension, the azotobacter chroococcum seed suspension, the lilac paecilomyces seed suspension, the Jingyang streptomyces seed suspension and the light gray streptomyces Hangzhou variant seed suspension comprises the following steps:
respectively selecting 1 ring of activated bacillus mucilaginosus, azotobacter chroococcum, paecilomyces lilacinus, streptomyces jingyangensis and streptomyces griseus Hangzhou varieties in the step (2) by using inoculating rings, and correspondingly inoculating the varieties into 5 triangular flasks filled with 150mL of liquid culture medium A, A, PDA of liquid culture medium, a first Gao's synthetic liquid culture medium and a first Gao's synthetic liquid culture medium for culture, wherein the bacillus mucilaginosus and the azotobacter chroococcum are subjected to shaking culture for 48 hours at the conditions of 150rpm and 25 ℃; shaking and culturing paecilomyces lilacinus at 150rpm and 25 deg.C for 7 days; shaking culturing Streptomyces jingyangensis for 7 days at 150rpm and 28 ℃; shaking and culturing the Hangzhou variety of the streptomyces lividans for 7 days at the temperature of 30 ℃ and at the speed of 150rpm by using a shaking table to obtain a bacillus mucilaginosus seed suspension, a azotobacter chroococcum seed suspension, a paecilomyces lilacinus seed suspension, a streptomyces jingyangensis seed suspension and a streptomyces lividans Hangzhou variety seed suspension respectively;
the method for preparing the bacillus mucilaginosus agent, the brown nodule azotobacter agent, the lilac paecilomyces agent, the Jingyang streptomyces agent and the light gray streptomyces Hangzhou variant agent comprises the following steps:
taking part of the pleurotus eryngii processing waste extracting solution obtained in the step (1), adjusting the pH value of the extracting solution to 7.4, and performing damp-heat sterilization at 122 ℃ for 10min to obtain a culture solution;
taking 5 triangular flasks with the volume of 250mL, and placing the obtained culture solution into the triangular flasks, wherein the volume of the culture solution in each triangular flask is 90 mL; then, respectively inoculating the bacillus mucilaginosus seed suspension, the azotobacter chroococcum seed suspension, the lilac paecilomyces seed suspension, the Jingyang streptomyces seed suspension and the light gray streptomyces Hangzhou variety seed suspension obtained in the step (3) into corresponding triangular flasks, wherein the inoculation amount in each triangular flask is 6%; after inoculation, carrying out shaking culture at the temperature of 29 ℃ and the rotating speed of 100rpm, wherein bacillus mucilaginosus and azotobacter chroococcum are cultured for 72h, and paecilomyces lilacinus, streptomyces jingyangensis and streptomyces griseus Hangzhou varieties are cultured for 9 days, so that the number of live bacteria of each microbial inoculum is 2-3 hundred million/mL, and thus the bacillus mucilaginosus microbial inoculum, the azotobacter chroococcum microbial inoculum, the paecilomyces lilacinus microbial inoculum, the streptomyces jingyangensis microbial inoculum and the streptomyces griseus Hangzhou variety microbial inoculum are obtained respectively;
the method for preparing the bacterial liquid in the step (5) comprises the following steps: taking part of the pleurotus eryngii processing waste extracting solution obtained in the step (1), and carrying out damp-heat sterilization on the extracting solution for 10min at 122 ℃; uniformly mixing the sterilized pleurotus eryngii processing waste extracting solution, a bacillus mucilaginosus microbial inoculum, a brown nodule azotobacter microbial inoculum, a lilac paecilomyces microbial inoculum, a Jingyang streptomyces microbial inoculum and a Streptomyces griseus Hangzhou variant microbial inoculum according to parts by weight to obtain the compound bacterial liquid.
Example four
The composite biological bacterial fertilizer special for the corn comprises the following components in parts by weight:
the composite bacteria liquid of the invention comprises 30 parts of composite bacteria liquid, 20 parts of bran, 15 parts of attapulgite, 40 parts of peat, 20 parts of bentonite and 25 parts of bean cakes.
The preparation method of the composite biological bacterial fertilizer special for the corns comprises the following steps:
preparing a compound bacterium liquid according to the method described in the second embodiment;
and II, crushing bran, attapulgite, bentonite and bean cakes, stirring and mixing the crushed bran, attapulgite, bentonite and bean cakes with peat and the prepared composite bacterial liquid, and drying the mixture at 25 ℃ to ensure that the water content of the mixture is 8% to obtain the composite biological bacterial fertilizer special for the corn.
EXAMPLE five
The composite biological bacterial fertilizer special for the corn comprises the following components in parts by weight:
the composite bacteria liquid of the invention comprises 35 parts of composite bacteria liquid, 18 parts of bran, 16 parts of attapulgite, 35 parts of peat, 22 parts of bentonite and 20 parts of bean cakes.
The preparation method of the composite biological bacterial fertilizer special for the corns comprises the following steps:
preparing a compound bacterium liquid according to the method described in the second embodiment;
and II, crushing the bran, the attapulgite, the bentonite and the bean cake, mixing the crushed mixture with peat and the prepared composite bacterial liquid under stirring, and drying the mixture at 28 ℃ to ensure that the water content of the mixture is 6% to obtain the composite biological bacterial fertilizer special for the corn.
EXAMPLE six
The composite biological bacterial fertilizer special for the corn comprises the following components in parts by weight:
the composite bacteria liquid of the invention comprises 40 parts of composite bacteria liquid, 15 parts of bran, 20 parts of attapulgite, 30 parts of peat, 25 parts of bentonite and 15 parts of bean cakes.
The preparation method of the composite biological bacterial fertilizer special for the corns comprises the following steps:
preparing a compound bacterium liquid according to the method described in the second embodiment;
and II, crushing bran, attapulgite, bentonite and bean cakes, stirring and mixing the crushed bran, attapulgite, bentonite and bean cakes with peat and the prepared composite bacterial liquid, and drying the mixture at 30 ℃ to ensure that the water content of the mixture is 5% to obtain the composite biological bacterial fertilizer special for the corn.
Example seven: determination of effect of composite biological bacterial fertilizer special for corn
In this example, the effect of the composite biological bacterial fertilizer special for tobacco corn obtained in the fifth example was measured.
The same plot is selected, divided into 5 blocks on average, and the 5 blocks are randomly divided into an experimental group, a control group 1, a control group 2, a control group 3 and a control group 4. Wherein, the experimental group applies the special composite biological bacterial fertilizer for tobacco corn obtained in the fifth embodiment; the control group 1 was applied with the greenling compound fertilizer; applying the compound fertilizer of the dragon Hao to the control group 2; a matrix fertilizer (which is prepared by mixing 18 parts of bran, 16 parts of attapulgite, 35 parts of peat, 22 parts of bentonite and 20 parts of bean cakes) is applied to the control group 3; control group 4 did not apply any fertilizer. Wherein the fertilization amounts and application times of the experimental group, the control group 1, the control group 2 and the control group 3 are the same, the effects of various fertilizers on the incidence rate of the corn northern leaf blight, the incidence rate of bacterial wilt, the incidence rate of stalk rot and the lodging resistance of corn are shown in the table 1; the influence of various fertilizers on the number of corn big and small spots, the plant height, the number of corn fruits and the yield is shown in table 2; the effect of various fertilizers on the nutrient content of corn is shown in table 3.
TABLE 1
| Treatment of | Northern leaf blight | Bacterial wilt | Stem rot of stalk | Degree of lodging | Rate of reverse turn |
| Experimental group | 0.0％ | 0.0％ | 0.0％ | 8° | 0.0％ |
| Control group 1 | 1.3％ | 2.3％ | 3.2％ | 12° | 0.2％ |
| Control group 2 | 2.5％ | 2.8％ | 3.5％ | 15° | 0.2％ |
| Control group 3 | 7.5％ | 10％ | 12％ | 25° | 0.3％ |
| Control group 4 | 9.5％ | 12％ | 15％ | 32° | 0.4％ |
TABLE 2
| Treatment of | The number of large and small spots/plant-1 | Plant height/cm | Corn fruit number/(number/strain) | Yield (kg/m)2) |
| Experimental group | 0 | 238 | 3.5 | 2.23 |
| Control group 1 | 12.7 | 183 | 2 | 1.78 |
| Control group 2 | 25.7 | 181 | 2 | 1.75 |
| Control group 3 | 56.2 | 176 | 1 | 1.63 |
| Control group 4 | 75.6 | 172 | 1 | 1.42 |
TABLE 3
As can be seen from the data in tables 1-3, the composite biological bacterial fertilizer special for tobacco corn can effectively promote the growth of corn plants, is beneficial to the transformation and absorption of corn to nutrient components in soil, increases the yield of corn, and can improve the disease resistance of corn.
It should be noted that the above mentioned embodiments are only preferred embodiments of the present invention, and it should be noted that those skilled in the art can make various changes, improvements and modifications without departing from the spirit of the present invention, and these changes, improvements and modifications should be construed as the protection scope of the present invention.
Claims (6)
1. The special composite biological bacterial fertilizer for the corn is characterized in that: the paint comprises the following components in parts by weight:
30-40 parts of composite bacteria liquid, 15-20 parts of bran, 15-20 parts of attapulgite, 30-40 parts of peat, 20-25 parts of bentonite and 15-25 parts of bean cakes;
the compound bacterium liquid is prepared from the following raw materials in parts by weight: 45-55 parts of pleurotus eryngii processing waste extracting solution, 15-20 parts of bacillus mucilaginosus microbial inoculum, 20-25 parts of azotobacter chroococcum microbial inoculum, 5-8 parts of lilac paecilomyces microbial inoculum, 5-10 parts of streptomyces jingyangensis microbial inoculum and 10-15 parts of streptomyces aegiensis Hangzhou variant microbial inoculum; the viable count of the bacillus mucilaginosus agent, the azotobacter chroococcum agent, the paecilomyces lilacinus agent, the streptomyces jingyangensis agent and the streptomyces griseus Hangzhou variant agent is 2-3 hundred million/mL;
the preparation method of the pleurotus eryngii processing waste extracting solution comprises the following steps:
cleaning mushroom feet and crushed mushrooms generated in the pleurotus eryngii processing process, crushing the mushroom feet and the crushed mushrooms into 40-50 meshes, adding water according to the weight ratio of 2:3 of the pleurotus eryngii to the water, heating the mixture to 90-98 ℃, decocting the mixture for 2-3 hours at the temperature, cooling and filtering the mixture, and collecting filtrate; centrifuging the collected filtrate at the rotating speed of 1500-; concentrating the obtained supernatant by falling film evaporation with a falling film evaporator until the refractive index of the solution reaches 35-45% to obtain a concentrated solution; then, adding 80-90% of ethanol into the concentrated solution, and modifying the concentrated solution, wherein the volume ratio of the added ethanol to the concentrated solution is 3: 2; filtering the denatured concentrated solution to remove precipitates generated after denaturation, and collecting filtrate; performing rotary evaporation on the filtrate to remove ethanol; and (3) performing tangential flow filtration on the solution obtained after rotary evaporation by using a filter membrane with the molecular weight cutoff of 5-10K, and collecting filtrate to obtain the pleurotus eryngii processing waste extracting solution.
2. The preparation method of the special composite biological bacterial fertilizer for corn as claimed in claim 1, characterized by comprising the following steps: it comprises the following steps:
i, preparing the compound bacterium liquid;
and II, crushing bran, attapulgite, bentonite and bean cakes, mixing with peat and the prepared composite bacterial liquid under stirring, and drying at 25-30 ℃ until the water content is 5-8% to obtain the composite biological bacterial fertilizer special for the corn.
3. The preparation method of the special composite biological bacterial fertilizer for corn according to claim 2, characterized by comprising the following steps: the preparation of the compound bacterium liquid in the step I comprises the following steps:
(1) preparing pleurotus eryngii processing waste extracting solution; (2) activating each strain; (3) preparing Bacillus mucilaginosus seed suspension, azotobacter chroococcum seed suspension, lilac paecilomyces seed suspension, streptomyces jingyangensis seed suspension, and streptomyces griseus Hangzhou variety seed suspension; (4) preparing Bacillus mucilaginosus agent, azotobacter nodosum agent, lilac paecilomyces agent, streptomyces jingyangensis agent and streptomyces griseus Hangzhou variant agent; (5) preparing a bacterial liquid;
wherein the method for preparing the bacillus mucilaginosus agent, the azotobacter chroococcum agent, the lilac paecilomyces agent, the Jingyang streptomyces agent and the Streptomyces griseus Hangzhou variant agent in the step (4) comprises the following steps of:
taking part of the pleurotus eryngii processing waste extracting solution obtained in the step (1), adjusting the pH value of the extracting solution to 7.2-7.4, and performing moist heat sterilization at the temperature of 121-;
taking 5 triangular flasks with the volume of 250mL, and placing the obtained culture solution into the triangular flasks, wherein the volume of the culture solution in each triangular flask is 80-90 mL; then, respectively inoculating the bacillus mucilaginosus seed suspension, the azotobacter chroococcum seed suspension, the lilac paecilomyces seed suspension, the Jingyang streptomyces seed suspension and the light gray streptomyces Hangzhou variety seed suspension obtained in the step (3) into corresponding triangular flasks, wherein the inoculation amount in each triangular flask is 4-6%; after inoculation, carrying out shaking culture at the conditions of 28-30 ℃ and the rotation speed of 100-250rpm, wherein bacillus mucilaginosus and azotobacter chroococcum are cultured for 48-72h, and paecilomyces lilacinus, streptomyces jingyangensis and streptomyces shallowlii are cultured for 7-9 days, so that the viable count of each microbial inoculum is 2-3 hundred million/mL, and thus the bacillus mucilaginosus microbial inoculum, the azotobacter chroococcum microbial inoculum, the paecilomyces lilacinus microbial inoculum, the streptomyces jingyangensis microbial inoculum and the streptomyces shallowlii Hangzhou microbial inoculum are respectively obtained;
the method for preparing the bacterial liquid in the step (5) comprises the following steps: taking part of the pleurotus eryngii processing waste extracting solution obtained in the step (1), and carrying out damp-heat sterilization on the extracting solution at the temperature of 121-; uniformly mixing the sterilized pleurotus eryngii processing waste extracting solution, a bacillus mucilaginosus microbial inoculum, a brown nodule azotobacter microbial inoculum, a lilac paecilomyces microbial inoculum, a Jingyang streptomyces microbial inoculum and a Streptomyces griseus Hangzhou variant microbial inoculum according to parts by weight to obtain the compound bacterial liquid.
4. The preparation method of the special composite biological bacterial fertilizer for corn according to claim 3, characterized by comprising the following steps: the method for preparing the pleurotus eryngii processing waste extracting solution in the step (1) comprises the following steps:
cleaning mushroom feet and crushed mushrooms generated in the pleurotus eryngii processing process, crushing the mushroom feet and the crushed mushrooms into 40-50 meshes, adding water according to the weight ratio of 2:3 of the pleurotus eryngii to the water, heating the mixture to 90-98 ℃, decocting the mixture for 2-3 hours at the temperature, cooling and filtering the mixture, and collecting filtrate; centrifuging the collected filtrate at the rotating speed of 1500-; concentrating the obtained supernatant by falling film evaporation with a falling film evaporator until the refractive index of the solution reaches 35-45% to obtain a concentrated solution; then, adding 80-90% of ethanol into the concentrated solution, and modifying the concentrated solution, wherein the volume ratio of the added ethanol to the concentrated solution is 3: 2; filtering the denatured concentrated solution to remove precipitate generated after denaturation, and collecting filtrate; performing rotary evaporation on the filtrate to remove ethanol; and (3) performing tangential flow filtration on the solution obtained after rotary evaporation by using a filter membrane with the molecular weight cutoff of 5-10K, and collecting filtrate to obtain the pleurotus eryngii processing waste extracting solution.
5. The preparation method of the special composite biological bacterial fertilizer for corn according to claim 3, characterized by comprising the following steps: the method for activating each strain in the step (2) comprises the following steps:
respectively inoculating bacillus mucilaginosus and azotobacter chroococcum on a slant containing a solid culture medium A, and separately culturing for 48-72h, wherein the culture temperature is controlled at 25-35 ℃;
wherein the solid medium A is K2HPO4、KH2PO4、MgSO4·7H2O、CaCO3·2H2O、Na2Mo·2H2O, yeast extract, mannitol, FeCl3And agar to prepare a culture medium;
inoculating Paecilomyces lilacinus to slant culture medium containing PDA slant culture medium, and culturing at 25-28 deg.C for 5-7 days;
inoculating Streptomyces jingyangensis on a slant containing Gaoshi synthetic first agar medium, and culturing for 5-7 days at 28-30 deg.C;
the Hangzhou variety of Streptomyces lividans is inoculated on a slant containing Gao's synthetic first agar medium and cultured for 5-7 days at a temperature controlled at 30 ℃.
6. The preparation method of the special composite biological bacterial fertilizer for corn according to claim 3, characterized by comprising the following steps: the method for preparing the bacillus mucilaginosus seed suspension, the azotobacter chroococcum seed suspension, the lilac paecilomyces seed suspension, the Jingyang streptomyces seed suspension and the light gray streptomyces Hangzhou variant seed suspension comprises the following steps:
selecting 1 ring of activated Bacillus mucilaginosus, azotobacter chroococcum, lilac paecilomyces and Jingyang from each inoculating ringStreptomyces and Hangzhou variety of Streptomyces griseus are respectively and correspondingly inoculated into 5 triangular flasks filled with 100 plus 150mL of liquid culture medium A, A, PDA liquid culture medium, Gao's synthetic first liquid culture medium and Gao's synthetic first liquid culture medium for culture, wherein, the Bacillus mucilaginosus and the azotobacter chroococcum are subjected to shaking culture for 24-48h under the conditions of 150 plus 200rpm and 25-35 ℃; culturing the lilac paecilomyces on a shaking table for 5-7 days at the temperature of 25-28 ℃ and 200rpm of 150-; shaking culturing Streptomyces jingyangensis for 5-7 days at 150-; shaking and culturing Hangzhou variety of Streptomyces lividans in a shaker at 150-; wherein the liquid culture medium A is K2HPO4、KH2PO4、MgSO4·7H2O、CaCO3·2H2O、Na2Mo·2H2O, yeast extract, mannitol, FeCl3Mixing to obtain the culture medium.
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| CN105130684A (en) * | 2015-09-29 | 2015-12-09 | 闽南师范大学 | Formula of compound biological bacterial fertilizer and preparation process and application thereof |
| CN107162673A (en) * | 2017-04-26 | 2017-09-15 | 闽南师范大学 | A kind of cultivation matrix of quickly breeding Banana Seedlings |
| CN109796274A (en) * | 2019-02-20 | 2019-05-24 | 福建三炬生物科技股份有限公司 | A kind of bio-feritlizer and its preparation method and application |
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| CN107162673A (en) * | 2017-04-26 | 2017-09-15 | 闽南师范大学 | A kind of cultivation matrix of quickly breeding Banana Seedlings |
| CN109796274A (en) * | 2019-02-20 | 2019-05-24 | 福建三炬生物科技股份有限公司 | A kind of bio-feritlizer and its preparation method and application |
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