CN114836351B - Compound microbial agent for killing insects and preventing diseases and biological fertilizer containing microbial agent - Google Patents
Compound microbial agent for killing insects and preventing diseases and biological fertilizer containing microbial agent Download PDFInfo
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
- C12N1/20—Bacteria; Culture media therefor
-
- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01N—PRESERVATION OF BODIES OF HUMANS OR ANIMALS OR PLANTS OR PARTS THEREOF; BIOCIDES, e.g. AS DISINFECTANTS, AS PESTICIDES OR AS HERBICIDES; PEST REPELLANTS OR ATTRACTANTS; PLANT GROWTH REGULATORS
- A01N63/00—Biocides, pest repellants or attractants, or plant growth regulators containing microorganisms, viruses, microbial fungi, animals or substances produced by, or obtained from, microorganisms, viruses, microbial fungi or animals, e.g. enzymes or fermentates
- A01N63/20—Bacteria; Substances produced thereby or obtained therefrom
-
- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01N—PRESERVATION OF BODIES OF HUMANS OR ANIMALS OR PLANTS OR PARTS THEREOF; BIOCIDES, e.g. AS DISINFECTANTS, AS PESTICIDES OR AS HERBICIDES; PEST REPELLANTS OR ATTRACTANTS; PLANT GROWTH REGULATORS
- A01N63/00—Biocides, pest repellants or attractants, or plant growth regulators containing microorganisms, viruses, microbial fungi, animals or substances produced by, or obtained from, microorganisms, viruses, microbial fungi or animals, e.g. enzymes or fermentates
- A01N63/20—Bacteria; Substances produced thereby or obtained therefrom
- A01N63/22—Bacillus
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- C—CHEMISTRY; METALLURGY
- C05—FERTILISERS; MANUFACTURE THEREOF
- C05F—ORGANIC FERTILISERS NOT COVERED BY SUBCLASSES C05B, C05C, e.g. FERTILISERS FROM WASTE OR REFUSE
- C05F3/00—Fertilisers from human or animal excrements, e.g. manure
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- C—CHEMISTRY; METALLURGY
- C05—FERTILISERS; MANUFACTURE THEREOF
- C05G—MIXTURES OF FERTILISERS COVERED INDIVIDUALLY BY DIFFERENT SUBCLASSES OF CLASS C05; MIXTURES OF ONE OR MORE FERTILISERS WITH MATERIALS NOT HAVING A SPECIFIC FERTILISING ACTIVITY, e.g. PESTICIDES, SOIL-CONDITIONERS, WETTING AGENTS; FERTILISERS CHARACTERISED BY THEIR FORM
- C05G3/00—Mixtures of one or more fertilisers with additives not having a specially fertilising activity
- C05G3/60—Biocides or preservatives, e.g. disinfectants, pesticides or herbicides; Pest repellants or attractants
-
- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02W—CLIMATE CHANGE MITIGATION TECHNOLOGIES RELATED TO WASTEWATER TREATMENT OR WASTE MANAGEMENT
- Y02W30/00—Technologies for solid waste management
- Y02W30/40—Bio-organic fraction processing; Production of fertilisers from the organic fraction of waste or refuse
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- Health & Medical Sciences (AREA)
- Zoology (AREA)
- General Health & Medical Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Organic Chemistry (AREA)
- Biotechnology (AREA)
- Pest Control & Pesticides (AREA)
- Wood Science & Technology (AREA)
- Microbiology (AREA)
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- Agronomy & Crop Science (AREA)
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Abstract
The invention relates to a compound microbial agent for disinsection and disease prevention and a biological fertilizer containing the microbial agent, which are obtained by mixing the compound microbial agent for disinsection and disease prevention and an organic fertilizer according to the mass ratio of 1: 1000-1100. The insecticidal disease-preventing compound microbial agent is obtained by mixing and adsorbing fermentation liquor of microorganisms and grass peat according to the mass ratio of 1: 3. The fermentation liquor of the microorganism comprises a Bacillus belgii ZLP-101 fermentation liquor and an additional bacteria fermentation liquor which are in a mass ratio of 50-60: 46-42; the additional bacteria comprise more than three of silicate bacteria K5, bacillus amyloliquefaciens D1, bacillus subtilis BSD-2, bacillus phosphate solubilizing P-6, bacillus brevis CH2-22 and Bacillus licheniformis BL-9. The invention is harmless to human and livestock, is environment-friendly, and can be widely applied to prevention and control of crop diseases and insect pests and increase production.
Description
Technical Field
The invention relates to the technical field of fertilizers, in particular to a compound microbial agent for killing insects and preventing diseases and a biological fertilizer containing the microbial agent.
Background
The chemical fertilizer and the chemical pesticide provide nutrient elements for crops to a certain extent, reduce the incidence rate of underground insect pests and indirectly improve the yield of the crops. However, with the increase of the cultivation period and the long-term accumulated application of fertilizer and pesticide, the drug resistance of underground pests is increased day by day, and the microbial flora structure related to nutrient circulation, organic matter decomposition and disease resistance is unbalanced, so that the number of soil-borne pathogenic bacteria and underground pests is increased, the soil nutrient is unbalanced, a series of problems of aggravation of diseases and insect pests, fertility reduction, physicochemical property change and the like occur, the yield of crops is reduced finally, the sustainable development of agriculture and food safety are seriously hindered, the bio-organic fertilizer which is environment-friendly, economical and effective and suitable for the crops is urgently needed, and the incidence rate of the diseases and insect pests of the crops can be reduced on the basis of effectively ensuring the high-quality supply of products.
The organic fertilizer contains rich organic matters, potassium and nutrient elements required by plants, can provide nutrition and living space for crops and microorganisms, and achieves the effects of slowly releasing nutrients and prolonging the shelf life of viable bacteria. The turf has the advantages of porosity, large surface area, large gaps in the structure of the turf, good hydrophilicity, capability of reducing soil hardness and promotion of better growth of crops.
Aiming at the technical current situation, if the resources can be fully utilized and the problems of soil-borne pests and soil-borne diseases of crops can be solved, the method has potential application prospect and wide development space.
Disclosure of Invention
The invention aims to provide a compound microbial agent for killing insects and preventing diseases and a biological fertilizer containing the microbial agent, which is harmless to human and livestock, is environment-friendly and can be widely applied to prevention and control of crop diseases and insect pests and increase of yield.
The invention adopts the following technical scheme:
a composite microbial agent for killing insects and preventing diseases comprises microbial fermentation liquor and grass peat, wherein the mass ratio of the microbial fermentation liquor to the grass peat is 1: 2.5-3; the microbial fermentation broth comprises a Bacillus belgii ZLP-101 fermentation broth.
Wherein the Bacillus belgii ZLP-101 is preserved in the common microorganism center of China Committee for culture Collection of microorganisms at the address of: the microbial research institute of China academy of sciences No. 3, xilu No. 1, beijing, chaoyang, china has a preservation date of 2020, 7 months and 3 days and a preservation number of CGMCC No.20130.
Furthermore, the number of the bacteria in the Bacillus belgii ZLP-101 fermentation liquor is more than or equal to 1.0 multiplied by 10 10 cfu/mL。
Further, the microbial fermentation liquid also comprises additional bacteria fermentation liquid, and the mass ratio of the Bacillus belgii ZLP-101 fermentation liquid to the additional bacteria fermentation liquid is 50-60: 42-75.
Further, the fermentation liquor of the additional bacteria comprises more than one of the following strains: silicate bacteria K5, bacillus amyloliquefaciens D1, bacillus subtilis BSD-2, bacillus phosphate solubilizing P-6, bacillus brevis CH2-22 and bacillus licheniformis BL-9.
Wherein the preservation number of the bacillus subtilis BSD-2 is CGMCC No.8440.
Wherein the silicate bacteria K5, the bacillus amyloliquefaciens D1, the bacillus phosphate solubilizing bacteria P-6, the bacillus brevis CH2-22 and the bacillus licheniformis BL-9 are provided by biological research institute of academy of sciences in Hebei province.
Wherein the number of the silicate bacteria K5 fermentation liquor is more than or equal to 2.0 multiplied by 10 8 cfu/mL, the number of bacteria of the bacillus amyloliquefaciens D1 fermentation liquid is more than or equal to 3.4 multiplied by 10 10 cfu/mL, the number of bacteria of the Bacillus subtilis BSD-2 fermentation liquor is more than or equal to 8.0 multiplied by 10 10 cfu/mL, the number of bacteria of the P-6 fermentation liquor of the phosphate-solubilizing bacillus is more than or equal to 3.0 multiplied by 10 9 cfu/mL, brevibacillus brevis CH2-22 fermentationThe number of bacteria in the solution is more than or equal to 1.0 multiplied by 10 10 cfu/mL, the number of bacteria of the bacillus licheniformis BL-9 fermentation liquor is more than or equal to 3.1 multiplied by 10 10 cfu/mL。
Preferably, the fermentation liquor of the additional bacteria comprises silicate bacteria K5 fermentation liquor, bacillus amyloliquefaciens D1 fermentation liquor and bacillus subtilis BSD-2 fermentation liquor in a mass ratio of 15: 20: 11.
Preferably, the fermentation liquor of the additional bacteria comprises a fermentation liquor of the phosphate-solubilizing bacillus P-6, a fermentation liquor of the Brevibacillus brevis CH2-22 and a fermentation liquor of the Bacillus licheniformis BL-9 in a mass ratio of 13: 10: 19.
Preferably, the fermentation liquor of the additional bacteria comprises fermentation liquor of bacillus subtilis BSD-2, fermentation liquor of bacillus phosphate solubilizing bacteria P-6, fermentation liquor of bacillus brevis CH2-22 and fermentation liquor of bacillus licheniformis BL-9 in a mass ratio of 28: 10: 25: 12.
The insecticidal disease-preventing bio-fertilizer containing the insecticidal disease-preventing compound microbial agent is prepared by mixing the insecticidal disease-preventing compound microbial agent and an organic fertilizer according to the mass ratio of 1: 1000-1100.
The organic fertilizer comprises the following components in parts by weight, and is obtained by composting and fermenting: 1 to 4 parts of animal waste, 2 to 2.5 parts of plant straw, 1.5 to 3 parts of flammulina velutipes mushroom bran and 1 to 2 parts of bran coat.
Wherein the animal manure comprises one or more of chicken manure, cow manure, pig manure, sheep manure and duck manure; the plant straw comprises one or more of wheat straw, corn straw, cotton straw, peanut straw, soybean straw, corncob, mung bean straw, rape straw, sweet potato straw, eggplant straw and sugarcane straw.
The flammulina velutipes mushroom residue is culture medium residues (flammulina velutipes mushroom residue) obtained after harvesting flammulina velutipes, and is commonly called edible mushroom cultivation waste or mushroom residues.
The biological fertilizer for killing insects and preventing diseases is prepared by the following steps:
(1) Respectively fermenting the microorganisms to respectively obtain zymocyte liquid of each microorganism;
(2) Mixing the zymophyte liquid to obtain mixed microbial liquid;
(3) Mixing the mixed microbial solution with turfy soil to obtain a compound microbial solution;
(4) Carrying out composting fermentation on organic fertilizer raw materials to obtain an organic fertilizer;
(5) And mixing the compound microbial bacteria liquid with an organic fertilizer to obtain the insecticidal disease-preventing biological fertilizer.
The invention has the beneficial effects that: the biological fertilizer has the effects of promoting the growth of crops such as cucumbers, corns, peanuts, wheat, potatoes, tomatoes and the like and improving the yield of the crops, can also prevent and control grubs, wireworms, cutworms, mole crickets and other underground insect pests, and can prevent and control cotton blight, cotton wilt, cotton black rot, cotton red rot, potato wilt, potato early blight, potato ring rot, corn leaf spot, corn sheath blight, wheat smut, peanut root rot, pepper bacterial wilt, cucumber wilt, tomato wilt and other fungal diseases. The biological fertilizer for killing pests and preventing diseases prepared by the invention is harmless to human and livestock, is environment-friendly, can be widely applied to the aspects of controlling crop pests and increasing yield, and has wide application prospect and development space.
Drawings
FIG. 1 is a photograph showing the colony morphology of Bacillus belgii ZLP-101 on LB solid medium.
FIG. 2 is a photograph showing the morphology of Bacillus belgii ZLP-101 cells on LB solid medium.
Detailed Description
The technical solution of the present invention is explained in detail below with reference to preferred embodiments. The following examples are only for illustrating and explaining the present invention and do not constitute a limitation to the technical solution of the present invention.
EXAMPLE 1 determination of the bacteriostatic Profile of the composite microbial sterile filtrate
Selecting cotton verticillium wilt, cotton black rot, potato early blight, potato wilt, potato ring rot, potato dry rot, tobacco brown spot, tomato wilt, cotton red rot, pepper ralstonia solanacearum, corn leaf spot, corn leaf blight, wheat smut, peanut root rot, grape white rot, tomato early blight, cucumber gray mold, cucumber powdery mildew, cucumber wilt and cucumber anthracnose as targets, and determining ZLP-101, D1 and P-6 by a plate confrontation method; ZLP-101, BSD-2, D1; ZLP-101, CH2-22, BL-9, K5, and 3 kinds of bacteria inhibiting spectrum of the composite microbe bacteria-free filtrate. The implementation data in table 1 show that the mixed sterile filtration of the functional microorganisms has better inhibition effect on different pathogenic bacteria, which indicates that the composite functional microorganism has wider inhibition spectrum, has inhibition effect on various types of pathogenic bacteria, and has good application effect in the field of prevention and treatment of crop diseases, especially soil-borne fungal diseases.
TABLE 1 inhibition of different pathogenic bacteria by sterile filtrates of complex-function microorganisms
Example 2
A biological fertilizer (formula 1) for killing insects and preventing diseases is prepared by the following steps:
(1) Respectively inoculating bacillus belgii ZLP-101 (with the preservation number of CGMCC No. 20130), silicate bacteria K5, bacillus amyloliquefaciens D1 and bacillus subtilis BSD-2 (with the preservation number of CGMCC No. 20129) on a solid medium by using a four-zone streaking method to culture a single colony; wherein the solid culture medium of the Bacillus beleisi ZLP-101, the Bacillus amyloliquefaciens D1 and the Bacillus subtilis BSD-2 is NB culture medium (5 g/L of beef extract, 10g/L of peptone, 5g/L of NaCl, 10g/L of glucose, 12.0g/L of agar and 7.1 +/-0.1 of pH), and is placed in a 37 ℃ incubator for culturing for 18-24h. Solid culture medium of silicate bacteria K5 (yeast extract powder 5.0g/L, calcium carbonate 0.1g/L, calcium chloride 0.04g/L, K) 2 HPO 4 ·3H 2 O 1.67g/L，KH 2 PO 4 0.87g/L，FeCl 3 ·6H 2 O 0.004g/L，MgSO 4 ·7H 2 O:0.1g/L, agar 15.0g/L, pH6.9 +/-0.1), and culturing in 25 deg.C incubator for 48-60 hr.
Then inoculating single colony into each 100ml liquid culture medium (except no agar, the other components are the same as solid culture medium) respectively, culturing bacillus belgii ZLP-101, bacillus amyloliquefaciens D1 and bacillus subtilis BSD-2 at 37 ℃ at 180r/min for 12-18h to logarithmic phase. Culturing silicate bacteria K5 at 25 deg.c for 12 hr to logarithmic phase at 180 r/min; finally, 100ml of liquid culture medium in the logarithmic phase is respectively inoculated into 800ml of liquid culture medium for culture, and finally zymogen liquid is obtained.
(2) The number of Bacillus belgii ZLP-101 in the zymocyte liquid is more than or equal to 1.0 multiplied by 10 10 cfu/mL, the number of silicate bacteria K5 is more than or equal to 7.0 multiplied by 10 9 cfu/mL, the number of bacillus amyloliquefaciens D1 is more than or equal to 3.4 multiplied by 10 10 cfu/mL, the number of bacillus subtilis BSD-2 is more than or equal to 8.0 multiplied by 10 10 cfu/mL。
(3) The composite microbial liquid is prepared by compounding Bacillus belgii ZLP-101, silicate bacteria K5, bacillus amyloliquefaciens D1 and Bacillus subtilis BSD-2 zymocyte liquid according to the mass ratio of 50: 15: 20: 11.
(4) And mixing and adsorbing the compound microbial inoculum and the grass carbon according to the mass ratio of 1: 3 to prepare the compound microbial inoculum.
(5) The chicken manure, the cotton straws, the needle mushroom fungus chaff and the rice chaff are subjected to compost fermentation according to the mass ratio of 4: 2.5: 1.5: 2, the compost is piled for about 30 days, the pile is turned for 1 time, water is timely supplemented according to the dry and wet conditions (the water content is kept between 50 and 60 percent), and the organic fertilizer can be made by decomposing in about 60 days at high temperature and high humidity in summer.
(6) The compound microbial agent and the organic fertilizer after compost fermentation are uniformly mixed according to the mass ratio of 1: 1000 to prepare the insecticidal disease-preventing biological fertilizer.
Example 3
A biological fertilizer (formula 2) for killing insects and preventing diseases is prepared by the following steps:
(1) Bacillus belgii ZLP-101, bacillus subtilis BSD-2 and phosphate-solubilizing sporeRespectively inoculating bacillus P-6, brevibacillus brevis CH2-22 and Bacillus licheniformis on a solid culture medium by using a four-zone streaking method to culture single colonies; wherein the solid culture medium of the Bacillus belgii ZLP-101, the Bacillus brevis CH2-22, the Bacillus subtilis BSD-2 and the Bacillus licheniformis is NB culture medium (5 g/L of beef extract, 10g/L of peptone, 5g/L of NaCl, 10g/L of glucose, 12.0g/L of agar and 7.1 +/-0.1 of pH), and is placed in an incubator at 37 ℃ for culturing for 18-24h. The solid culture medium of the phosphate-solubilizing bacillus P-6 is an inorganic phosphate culture medium (sucrose 2g/L, glucose 2g/L, NH) 4 Cl 1.5g/L、KCl 0.3g/L、MgSO 4 .7H 2 O0.4 g/L, naCl 0.2g/L, calcium phosphate 20g/L, pH 7.0), and culturing in 32 deg.C incubator for 20-24 hr.
Then inoculating the single colonies into 100ml of liquid culture medium (except no agar, the other components are the same as the solid culture medium); bacillus belgii ZLP-101, bacillus phosphate solubilizing P-6, bacillus subtilis BSD-2, bacillus brevis CH2-22 and Bacillus licheniformis are cultured at the temperature of 37 ℃ at 180r/min for 12-18h until logarithmic phase. Finally, 100ml of liquid culture medium in the logarithmic phase is respectively inoculated into 800ml of liquid culture medium for culture, and finally zymogen liquid is obtained.
(2) Finally, the number of the Bacillus belgii ZLP-101 in the zymocyte liquid is more than or equal to 1.0 multiplied by 10 10 cfu/mL, the bacterial count of the bacillus subtilis BSD-2 fermentation liquor is more than or equal to 8.0 multiplied by 10 10 cfu/mL, the number of bacteria of the P-6 fermentation liquor of the phosphate-solubilizing bacillus is more than or equal to 3.0 multiplied by 10 9 cfu/mL, the number of bacteria in the Brevibacillus brevis CH2-22 fermentation liquor is more than or equal to 1.0 multiplied by 10 10 cfu/mL, the number of bacteria of the fermentation liquor of the bacillus licheniformis BL-9 is more than or equal to 3.1 multiplied by 10 10 cfu/mL。
(3) The composite microbial liquid is prepared by compounding fermentation liquids of Bacillus belgii ZLP-101, bacillus subtilis BSD-2, bacillus phosphate solubilizing bacteria P-6, bacillus brevis CH2-22 and Bacillus licheniformis BL-9 according to the mass ratio of 45: 28: 10: 25: 12.
(4) And adsorbing the compound microbial inoculum and the grass carbon according to the mass ratio of 1: 3 to prepare the compound microbial inoculum.
(5) The cow dung, the corn straws, the needle mushroom fungus chaff and the bran coat are subjected to compost fermentation according to the ratio of 2: 2.5: 3: 1, the compost is piled for about 30 days, the pile is turned for 1 time, water is supplemented in time according to the dry and wet conditions (the water content is kept between 50 and 60 percent), and the organic fertilizer can be prepared by decomposing in about 60 days under high temperature and high humidity in summer.
(6) The compound microbial agent and the organic fertilizer after additional fertilization and fermentation are uniformly mixed according to the mass ratio of 1: 1200, and the insecticidal disease-preventing biological fertilizer is prepared.
Example 4
A biological fertilizer (formula 3) for killing insects and preventing diseases is prepared by the following steps:
(1) Respectively inoculating and culturing a single colony of Bacillus belgii ZLP-101, bacillus phosphate solubilizing P-6, bacillus brevis CH2-22 and Bacillus licheniformis BL-9 on a solid culture medium by using a four-zone streaking method; wherein the solid culture medium of the Bacillus belgii ZLP-101, the Bacillus brevis CH2-22 and the Bacillus licheniformis BL-9 is NB culture medium (5 g/L of beef extract, 10g/L of peptone, 5g/L of NaCl, 10g/L of glucose, 12.0g/L of agar and 7.1 +/-0.1 of pH), and is placed in a 37 ℃ incubator for culturing for 48h. The solid culture medium of the phosphate-solubilizing bacillus P-6 is an inorganic phosphate culture medium (sucrose 2g/L, glucose 2g/L, NH) 4 Cl 1.5g/L、KCl 0.3g/L、MgSO 4 .7H 2 O0.4 g/L, naCl 0.2g/L, calcium phosphate 20g/L, pH 7.0), and culturing in 32 deg.C incubator for 20-24 hr.
Then inoculating the single colony into 100ml liquid culture medium (except no agar, the other components are the same as the solid culture medium) respectively, culturing for 12-18h to logarithmic phase by using Bacillus belgii ZLP-101 (37 ℃), bacillus phosphate solubilizing P-6 (32 ℃), bacillus brevis CH2-22 (37 ℃) and Bacillus licheniformis BL-9 (37 ℃) at 180 r/min. Finally, 100ml of liquid culture medium in the logarithmic phase is respectively inoculated into 800ml of liquid culture medium for culture, and finally zymogen liquid is obtained.
(2) The number of Bacillus belgii ZLP-101 in the zymocyte liquid is more than or equal to 1.0 multiplied by 10 10 cfu/mL, the number of the bacillus phosphate solubilizing P-6 is more than or equal to 3.0 multiplied by 10 9 cfu/mL, the number of the Brevibacillus brevis CH2-22 is more than or equal to 1.0 multiplied by 10 10 The number of cfu/mL and the number of bacillus licheniformis BL-9 is more than or equal to 3.1 multiplied by 10 10 cfu/mL。
(3) The composite microbial liquid is prepared by compounding fermentation liquid of Bacillus belgii ZLP-101, bacillus phosphate solubilizing P-6, bacillus brevis CH2-22 and Bacillus licheniformis BL-9 according to the mass ratio of 60: 13: 10: 19.
(4) Adsorbing the compound microbial bacteria liquid and the grass carbon according to the mass ratio of 1: 3 to prepare the compound microbial bacteria agent.
(5) Carrying out compost fermentation on pig manure, wheat straws, needle mushroom fungus chaff and bran coat according to the ratio of 1: 2: 2.5: 1.5, piling the compost for about 30 days, turning the compost for 1 time, supplementing water in time according to the dry and wet conditions (keeping the water content between 50 and 60 percent), and carrying out high-temperature and high-humidity treatment in summer for about 60 days to become thoroughly decomposed into the organic fertilizer.
(6) The compound microbial agent and the organic fertilizer after compost fermentation are uniformly mixed according to the mass ratio of 1: 1100, and the insecticidal disease-preventing biological fertilizer is prepared.
Example 5
A biological fertilizer (formula 4) for killing insects and preventing diseases is prepared by the following steps:
(1) Inoculating Bacillus belgii ZLP-101 (with the preservation number of CGMCC No. 20130) on a solid culture medium by using a four-region streaking method to culture a single colony; wherein the solid culture medium is NB culture medium (beef extract 5g/L, peptone 10g/L, naCl 5g/L, glucose 10g/L, agar 12.0g/L, pH 7.1 + -0.1), and is placed in a 37 deg.C incubator for culturing for 18-24h.
Then inoculating a single colony into 100ml of liquid culture medium (except no agar, the other components are the same as the solid culture medium) respectively, and culturing the Bacillus belgii ZLP-101 at the temperature of 37 ℃ for 12-18h to a logarithmic phase. Finally, 100ml of liquid culture medium in the logarithmic phase is respectively inoculated into 800ml of liquid culture medium for culture, and finally zymogen liquid is obtained.
(2) The number of Bacillus belgii ZLP-101 in the zymocyte liquid is more than or equal to 1.0 multiplied by 10 10 cfu/mL。
(3) And mixing and adsorbing the compound microbial bacteria liquid and the grass peat according to the mass ratio of 1: 3 to prepare the compound microbial bacteria agent.
(4) The chicken manure, the cotton straws, the needle mushroom fungus chaff and the rice chaff are subjected to compost fermentation according to the mass ratio of 4: 2.5: 1.5: 2, the compost is piled for about 30 days, the pile is turned for 1 time, water is timely supplemented according to the dry and wet conditions (the water content is kept between 50 and 60 percent), and the organic fertilizer can be made by decomposing in about 60 days at high temperature and high humidity in summer.
(5) The compound microbial agent and the organic fertilizer after compost fermentation are uniformly mixed according to the mass ratio of 1: 1000 to prepare the insecticidal disease-preventing biological fertilizer.
Example 6 pesticidal effect of the composite microbial fermentation sterile filtrate on grub and other pests
B, mixing Bacillus belgii ZLP-101, silicate bacteria K5, bacillus amyloliquefaciens D1 and Bacillus subtilis BSD-2 (the mixture ratio of fermentation liquor is shown in formula 1); bacillus belgii ZLP-101, bacillus subtilis BSD-2, bacillus phosphate solubilizing P-6, bacillus brevis CH2-22 and Bacillus licheniformis BL-9 (the proportion of fermentation liquor is shown in formula 2); bacillus belgii ZLP-101, bacillus phosphate solubilizing P-6, bacillus brevis CH2-22 and Bacillus licheniformis BL-9 (the proportion of fermentation liquor is shown in formula 3); bacillus belgii ZLP-101 fermentation broth (formula 4); and mixing fermentation liquor obtained by fermenting each strain according to the formula, and filtering to obtain sterile filtrate of each formula. Diluting the raw solution by 20 times, 50 times, 100 times and 200 times to prepare medicinal soil, adding appropriate amount of foodstuff (crushed corn stalk, etc.), mixing, and preparing medicinal soil with clear water as control. Selecting grubs (larvae of the white star scarab), diamond back moths, beet armyworms, prodenia litura, cabbage caterpillars, cicadas and cockroaches with the same size, and repeating the treatment for 3 times in an environment with the relative humidity of 50-70% and the temperature of 25-28 ℃. The number of test insects per treatment is 30, and the test insects are repeated for 3 times. And (4) inoculating the insects, culturing at room temperature, observing every 8 hours, and counting the death rate after 56 hours. As can be seen from the implementation data in Table 2, the composite microbial fermentation product in the biological fertilizer for killing insects and preventing diseases has good killing effect on pests such as grubs and the like.
TABLE 2 effect of the composite microbial fermentation product of the present invention on killing grubs and other pests
Example 7 quality determination of pesticidal disease-preventing biofertilizer
The detection results in the table 3 show that the quality of the insecticidal disease-preventing biological fertilizer meets the requirements of agricultural industry standard NY884-2012 of the people's republic of China.
TABLE 3 quality test results of the insecticidal and disease-preventing biofertilizer of the present invention
Example 8 yield increase effect of pesticidal disease-preventing biofertilizer on field crops and vegetables
The insecticidal disease-preventing biological fertilizer (formula 1, formula 2, formula 3 and formula 4) is applied to corn, peanut and wheat field crops, cucumber, tomato and potato vegetables, and 150g of the insecticidal disease-preventing biological fertilizer is applied to 1 group. The treatment 2 groups are common biological organic fertilizers (root-force multi-biological organic fertilizer) on the market, purchased from root-force multi-biological science and technology limited company and applied by 100-200g per square. The control group is applied with chemical fertilizer group (NPK organic ternary compound fertilizer 16-16-16) purchased from Jinduobao Biotechnology Co., ltd, and applied with about 50g per square, and uniformly spread or buried 15cm under soil, far away from the root. Different crops are applied in different amounts. The yields of various crops and vegetables of each group of 4 formulas are counted, and as can be seen from the yields of each group in table 4, the organic fertilizer is superior to the existing biological organic fertilizer on the market in the aspect of improving the crop yield, wherein the yields of corn, peanut, wheat, cucumber, tomato and potato are respectively improved by 6.76% -17.51%, 12.64% -19.90%, 7.29% -9.71%, 2.25% -6.28%, 6.05% -9.30% and 3.72% -6.14% compared with the common biological organic fertilizer, and the biological organic fertilizer has significant differences. Compared with chemical fertilizers, the yield has no significant difference, which shows that the insecticidal disease-preventing biological fertilizer has the potential of replacing the chemical fertilizers.
TABLE 4 Effect of different treatments on crops such as maize, peanut, wheat, cucumber, tomato
Note: the same lower case letters in the same row indicate no significant difference, P > 0.05. Different lower case letters indicate significant differences, P < 0.05. The same applies below.
Example 9 prevention of soil-borne diseases in greenhouse vegetables by insecticidal and disease-preventing biofertilizer
The application of the insecticidal disease-preventing bio-fertilizer (formula 1, formula 2, formula 3 and formula 4) is respectively carried out in greenhouse for planting field crops such as potatoes, corns, wheat and peanuts and vegetables such as hot peppers, cucumbers and tomatoes, 150g of the insecticidal disease-preventing bio-fertilizer is applied to a treatment group per square, about 50g of chemical fertilizer (nitrogen-phosphorus-potassium organic ternary compound fertilizer 16-16-16) is applied to a control group per square, and the insecticidal disease-preventing bio-fertilizer is uniformly spread or buried 15cm under soil and is far away from roots.
Through recording and statistics of the disease conditions of various crops and vegetables, the control effects of the treatment group of the formula 1 on potato wilt, potato early blight, potato ring rot, corn leaf spot, corn sheath blight, wheat smut, peanut root rot, pepper bacterial wilt, cucumber wilt and tomato wilt are increased by 67.8%, 63.62%, 64.05%, 58.47%, 67.30%, 58.80%, 62.91%, 57.05%, 69.82% and 68.12% compared with the control group. The control effect of the treatment group of the formula 2 on potato wilt, potato early blight, potato ring rot, corn leaf spot, corn sheath blight, wheat smut, peanut root rot, pepper bacterial wilt, cucumber wilt and tomato wilt is increased by 61.09%, 60.55%, 71.78%, 62.76%, 65.99%, 59.63%, 67.02%, 69.09%, 68.34% and 66.20% compared with the control group. The control effect of the treatment group of the formula 3 on potato wilt, potato early blight, potato ring rot, corn leaf spot, corn sheath blight, wheat smut, peanut root rot, pepper bacterial wilt, cucumber wilt and tomato wilt is increased by 58.32%, 53.65%, 53.80%, 69.43%, 68.55%, 55.40%, 70.26%, 67.24%, 68.44% and 66.44% compared with the control group. The control effect of the treatment group of the formula 4 on potato wilt, potato early blight, potato ring rot, corn leaf spot, corn sheath blight, wheat smut, peanut root rot, pepper bacterial wilt, cucumber wilt and tomato wilt is increased by 57.40%, 49.23%, 49.20%, 60.39%, 65.46%, 54.40%, 60.84%, 61.07%, 65.81% and 60.24% compared with the control group. The implementation result shows that the insecticidal disease-preventing bio-fertilizer has good control effect on soil-borne diseases of field crops and facility vegetables.
TABLE 5 prevention of soil-borne diseases of vegetables such as eggplant, lettuce and hot pepper by different treatments
Example 10 control of soil pests with insecticidal and disease-preventive biofertilizers
The method is implemented on corn, peanut and potato fields respectively, and compared with chemical fertilizers (16-16-16 of nitrogen-phosphorus-potassium organic ternary compound fertilizers) applied to the bottom, about 50g of the chemical fertilizers are applied to each square, and the chemical fertilizers are uniformly spread or buried 15cm under the soil and far away from roots. The insecticide disease-prevention biofertilizer (formula 3) is applied to the bottom of the treatment layer, and 150g of the insecticide disease-prevention biofertilizer is applied to each square. Each crop or vegetable is divided into 12 cells, and the area of each cell is 20m 2 At the same timeAnd arranging the machine blocks. And (5) investigating all plants in each cell, and calculating the seedling protection rate and the control effect.
Seedling retention (%) = (total number of plants-number of damaged plants)/total number of plants × 100.
Control effect (%) = (treatment seedling protection rate-CK seedling protection rate)/(1-CK seedling protection rate) × 100.
TABLE 6 controlling effect of the present invention on cutworm, wireworm and grub underground pests
According to the implementation data in table 6, the control effects of the insecticidal disease-prevention bio-fertilizer on the cutworms in peanut fields, the wireworms in corn fields and the white grubs in potato fields are 75.56%, 69.23% and 77.16%, respectively, which shows that the insecticidal disease-prevention bio-fertilizer has remarkable control effects on the soil insects of peanuts, corns and potatoes.
Claims (5)
1. The compound microbial agent for killing insects and preventing diseases is characterized by comprising microbial fermentation liquor and grass peat, wherein the mass ratio of the microbial fermentation liquor to the grass peat is 1: 2.5-3; the microbial fermentation broth is a Bacillus beiLeisi ZLP-101 fermentation broth; the preservation number of the Bacillus belgii ZLP-101 is CGMCC No.20130.
2. The compound microbial inoculant for killing insects and preventing diseases as claimed in claim 1, wherein the number of bacteria in the fermentation broth of Bacillus belgii ZLP-101 is not less than 1.0 x 10 10 cfu/mL。
3. An insecticidal disease-prevention biofertilizer containing the insecticidal disease-prevention compound microbial agent as claimed in claim 1 or 2, characterized in that the insecticidal disease-prevention compound microbial agent and organic fertilizer are mixed according to the mass ratio of 1: 1000-1100 to obtain the insecticidal disease-prevention biofertilizer.
4. The insecticidal disease-preventing biofertilizer according to claim 3, characterized in that the organic fertilizer comprises the following components in parts by weight which are obtained by composting and fermentation: 1 to 4 parts of animal waste, 2 to 2.5 parts of plant straw, 1.5 to 3 parts of flammulina velutipes mushroom bran and 1 to 2 parts of bran coat.
5. The insecticidal disease-preventing biofertilizer according to claim 4, characterized in that it is prepared by the following steps:
(1) Fermenting the microorganisms to obtain a zymophyte liquid of the microorganisms;
(2) Mixing the zymophyte liquid with turfy soil to obtain a compound microorganism bacterium liquid;
(3) Carrying out composting fermentation on organic fertilizer raw materials to obtain an organic fertilizer;
(4) And mixing the compound microbial bacteria liquid with an organic fertilizer to obtain the insecticidal disease-preventing biological fertilizer.
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