CN117603880A - Microbial agent for preventing and treating gummy stem blight of melon and preparation method thereof - Google Patents
Microbial agent for preventing and treating gummy stem blight of melon and preparation method thereof Download PDFInfo
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- CN117603880A CN117603880A CN202311658214.3A CN202311658214A CN117603880A CN 117603880 A CN117603880 A CN 117603880A CN 202311658214 A CN202311658214 A CN 202311658214A CN 117603880 A CN117603880 A CN 117603880A
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- 235000015510 Cucumis melo subsp melo Nutrition 0.000 title claims abstract description 54
- FJJCIZWZNKZHII-UHFFFAOYSA-N [4,6-bis(cyanoamino)-1,3,5-triazin-2-yl]cyanamide Chemical compound N#CNC1=NC(NC#N)=NC(NC#N)=N1 FJJCIZWZNKZHII-UHFFFAOYSA-N 0.000 title claims abstract description 47
- 230000000813 microbial effect Effects 0.000 title claims abstract description 39
- 238000002360 preparation method Methods 0.000 title claims abstract description 17
- 241000219112 Cucumis Species 0.000 title claims abstract 9
- 241000894006 Bacteria Species 0.000 claims abstract description 49
- 239000002131 composite material Substances 0.000 claims abstract description 39
- 239000003795 chemical substances by application Substances 0.000 claims abstract description 37
- 150000004676 glycans Chemical class 0.000 claims abstract description 27
- 229920001282 polysaccharide Polymers 0.000 claims abstract description 27
- 239000005017 polysaccharide Substances 0.000 claims abstract description 27
- CSNNHWWHGAXBCP-UHFFFAOYSA-L Magnesium sulfate Chemical compound [Mg+2].[O-][S+2]([O-])([O-])[O-] CSNNHWWHGAXBCP-UHFFFAOYSA-L 0.000 claims abstract description 22
- 241000193744 Bacillus amyloliquefaciens Species 0.000 claims abstract description 18
- 241000223260 Trichoderma harzianum Species 0.000 claims abstract description 18
- 241000589540 Pseudomonas fluorescens Species 0.000 claims abstract description 17
- VYPSYNLAJGMNEJ-UHFFFAOYSA-N Silicium dioxide Chemical compound O=[Si]=O VYPSYNLAJGMNEJ-UHFFFAOYSA-N 0.000 claims abstract description 11
- 229910000365 copper sulfate Inorganic materials 0.000 claims abstract description 11
- ARUVKPQLZAKDPS-UHFFFAOYSA-L copper(II) sulfate Chemical compound [Cu+2].[O-][S+2]([O-])([O-])[O-] ARUVKPQLZAKDPS-UHFFFAOYSA-L 0.000 claims abstract description 11
- 235000003891 ferrous sulphate Nutrition 0.000 claims abstract description 11
- 239000011790 ferrous sulphate Substances 0.000 claims abstract description 11
- BAUYGSIQEAFULO-UHFFFAOYSA-L iron(2+) sulfate (anhydrous) Chemical compound [Fe+2].[O-]S([O-])(=O)=O BAUYGSIQEAFULO-UHFFFAOYSA-L 0.000 claims abstract description 11
- 229910000359 iron(II) sulfate Inorganic materials 0.000 claims abstract description 11
- 229910052943 magnesium sulfate Inorganic materials 0.000 claims abstract description 11
- 235000019341 magnesium sulphate Nutrition 0.000 claims abstract description 11
- OTYBMLCTZGSZBG-UHFFFAOYSA-L potassium sulfate Chemical compound [K+].[K+].[O-]S([O-])(=O)=O OTYBMLCTZGSZBG-UHFFFAOYSA-L 0.000 claims abstract description 11
- 229910052939 potassium sulfate Inorganic materials 0.000 claims abstract description 11
- 235000011151 potassium sulphates Nutrition 0.000 claims abstract description 11
- 235000015393 sodium molybdate Nutrition 0.000 claims abstract description 11
- 239000011684 sodium molybdate Substances 0.000 claims abstract description 11
- TVXXNOYZHKPKGW-UHFFFAOYSA-N sodium molybdate (anhydrous) Chemical compound [Na+].[Na+].[O-][Mo]([O-])(=O)=O TVXXNOYZHKPKGW-UHFFFAOYSA-N 0.000 claims abstract description 11
- NWONKYPBYAMBJT-UHFFFAOYSA-L zinc sulfate Chemical compound [Zn+2].[O-]S([O-])(=O)=O NWONKYPBYAMBJT-UHFFFAOYSA-L 0.000 claims abstract description 11
- 229910000368 zinc sulfate Inorganic materials 0.000 claims abstract description 11
- 229960001763 zinc sulfate Drugs 0.000 claims abstract description 11
- 239000002068 microbial inoculum Substances 0.000 claims abstract description 4
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- 235000017060 Arachis glabrata Nutrition 0.000 claims description 12
- 244000105624 Arachis hypogaea Species 0.000 claims description 12
- 235000010777 Arachis hypogaea Nutrition 0.000 claims description 12
- 235000018262 Arachis monticola Nutrition 0.000 claims description 12
- 235000012054 meals Nutrition 0.000 claims description 12
- 235000020232 peanut Nutrition 0.000 claims description 12
- 238000010298 pulverizing process Methods 0.000 claims description 10
- 241001655322 Streptomycetales Species 0.000 claims description 6
- 239000007788 liquid Substances 0.000 claims description 5
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- 230000000694 effects Effects 0.000 abstract description 18
- 230000005764 inhibitory process Effects 0.000 abstract description 9
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- 150000001875 compounds Chemical class 0.000 description 11
- 241000187391 Streptomyces hygroscopicus Species 0.000 description 9
- 241000196324 Embryophyta Species 0.000 description 6
- 235000009847 Cucumis melo var cantalupensis Nutrition 0.000 description 4
- 239000013043 chemical agent Substances 0.000 description 3
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- 238000012360 testing method Methods 0.000 description 2
- LDXJRKWFNNFDSA-UHFFFAOYSA-N 2-(2,4,6,7-tetrahydrotriazolo[4,5-c]pyridin-5-yl)-1-[4-[2-[[3-(trifluoromethoxy)phenyl]methylamino]pyrimidin-5-yl]piperazin-1-yl]ethanone Chemical compound C1CN(CC2=NNN=C21)CC(=O)N3CCN(CC3)C4=CN=C(N=C4)NCC5=CC(=CC=C5)OC(F)(F)F LDXJRKWFNNFDSA-UHFFFAOYSA-N 0.000 description 1
- UHPMCKVQTMMPCG-UHFFFAOYSA-N 5,8-dihydroxy-2-methoxy-6-methyl-7-(2-oxopropyl)naphthalene-1,4-dione Chemical compound CC1=C(CC(C)=O)C(O)=C2C(=O)C(OC)=CC(=O)C2=C1O UHPMCKVQTMMPCG-UHFFFAOYSA-N 0.000 description 1
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- MKYBYDHXWVHEJW-UHFFFAOYSA-N N-[1-oxo-1-(2,4,6,7-tetrahydrotriazolo[4,5-c]pyridin-5-yl)propan-2-yl]-2-[[3-(trifluoromethoxy)phenyl]methylamino]pyrimidine-5-carboxamide Chemical compound O=C(C(C)NC(=O)C=1C=NC(=NC=1)NCC1=CC(=CC=C1)OC(F)(F)F)N1CC2=C(CC1)NN=N2 MKYBYDHXWVHEJW-UHFFFAOYSA-N 0.000 description 1
- OAICVXFJPJFONN-UHFFFAOYSA-N Phosphorus Chemical compound [P] OAICVXFJPJFONN-UHFFFAOYSA-N 0.000 description 1
- ZLMJMSJWJFRBEC-UHFFFAOYSA-N Potassium Chemical compound [K] ZLMJMSJWJFRBEC-UHFFFAOYSA-N 0.000 description 1
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- JAWMENYCRQKKJY-UHFFFAOYSA-N [3-(2,4,6,7-tetrahydrotriazolo[4,5-c]pyridin-5-ylmethyl)-1-oxa-2,8-diazaspiro[4.5]dec-2-en-8-yl]-[2-[[3-(trifluoromethoxy)phenyl]methylamino]pyrimidin-5-yl]methanone Chemical compound N1N=NC=2CN(CCC=21)CC1=NOC2(C1)CCN(CC2)C(=O)C=1C=NC(=NC=1)NCC1=CC(=CC=C1)OC(F)(F)F JAWMENYCRQKKJY-UHFFFAOYSA-N 0.000 description 1
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- 239000011591 potassium Substances 0.000 description 1
- 229910052700 potassium Inorganic materials 0.000 description 1
- 235000007686 potassium Nutrition 0.000 description 1
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- 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
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- 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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- 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
- A01N63/27—Pseudomonas
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- 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
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- A01N63/20—Bacteria; Substances produced thereby or obtained therefrom
- A01N63/28—Streptomyces
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- 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
- A01N63/30—Microbial fungi; Substances produced thereby or obtained therefrom
- A01N63/38—Trichoderma
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- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01P—BIOCIDAL, PEST REPELLANT, PEST ATTRACTANT OR PLANT GROWTH REGULATORY ACTIVITY OF CHEMICAL COMPOUNDS OR PREPARATIONS
- A01P3/00—Fungicides
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Abstract
The invention relates to a microbial agent for preventing and treating melon gummy stem blight and a preparation method thereof, belonging to the technical field of agricultural planting disease prevention and treatment. The microbial agent disclosed by the invention comprises, by weight, 10-15 parts of biological composite bacteria, 20-30 parts of diatomite, 40-50 parts of algal polysaccharide, 3-5 parts of zinc sulfate, 0.5-1 part of copper sulfate, 1-3 parts of sodium molybdate, 0.2-0.5 part of ferrous sulfate, 1-2 parts of potassium sulfate and 0.05-0.1 part of magnesium sulfate; wherein the biological composite bacteria comprise non-hydroscopic streptomyces, bacillus amyloliquefaciens, trichoderma harzianum and pseudomonas fluorescens. The microbial inoculum has good effect on preventing and treating melon gummy stem blight, has good inhibition effect on gummy stem blight germs, obviously reduces the incidence rate of melon gummy stem blight, and realizes the yield and income increase of melons.
Description
Technical Field
The invention belongs to the technical field of agricultural planting disease control, and relates to a microbial agent for controlling melon gummy stem blight and a preparation method thereof.
Background
The melon is annual creeping or climbing herbal plant of Cucurbitaceae melon. The stems and branches have edges, and have brown or white rough bristles and wart. Tendrils are fine, single and slightly soft. The length of the leaf stalk is 8-12 cm, and the leaf stalk is provided with grooves and short bristles. Leaf blades are thick paper, nearly circular or kidney-shaped. The plant is cultivated widely in india and China in the world from temperate to tropical regions. Gardening is divided into tens of strains, for example, common muskmelon, hami melon, and white melon all belong to different strains. The whole herb has the effects of eliminating inflammation, removing toxic materials, promoting vomiting, removing dampness, and eliminating jaundice.
Melon is one of ten fruits in the world, is rich in nutrition, is fragrant, sweet and delicious, is popular with people, and has higher planting economic benefit. However, the disease is often suffered during the production and planting process to limit the high quality and high yield of the melons, wherein gummy stem blight is a common and serious disease among diseases, the damage degree is higher than that of fusarium wilt and epidemic disease, once the gummy stem is spread, the control is difficult, and especially in continuous cropping and greenhouses, the yield and quality of the melons are often severely reduced.
Gummy stem blight, also known as "black rot," mainly occurs in open-field rainy seasons. The melon gummy stem blight can occur in the whole growth period, mainly is a melon vine, can also be a leaf and a fruit, is ill in the seedling period, the stem part of the leaf has small water stain spots, gradually radiates to the periphery to form a yellow brown focus, and seriously causes the disease to be crumpled and fatal; when the true leaves are ill, yellow brown to blackish brown irregular disease spots exist at the initial stage, when the leaf is wet, the disease spots rapidly spread to the whole leaves, the leaf edges are damaged and then form blackish brown arcs, black small particles are arranged on the surfaces of dead leaves, and the leaves are always star-shaped broken after being dried; the melon vine is infected, the proximal section is in a light yellow oily stain shape and is oval to irregular disease spots, the disease spots are slightly concave, small black spots are densely grown on the spots, the epidermis is broken when serious, a yellowish white jelly is secreted, and the dried sections are in a reddish brown or black block shape; fruit body is infected, water stain-like disease spots appear at first, then the center becomes brown dead spots, the spots are star-shaped and cracked, the inside of the spots is in a wood plug shape, and the spots are blackened and then decayed.
Pathogenic bacteria of gummy stem blight mainly adopt conidium, cord hypha and ascus shells to fall into soil and insufficiently decomposed manure along with disease residues for overwintering, and seeds can also carry bacteria with the bacterial carrying rate of 5% -30%, and can survive for more than one year in the seeds; conidia are formed on the affected plant body, spread and infest in modes of wind, rain, irrigation water, farm work operation and the like, invade from air holes, water holes or wounds, and repeatedly infest to cause spreading hazard.
The occurrence degree of the disease is closely related to the temperature, humidity, cultivation management and the like of small weather of a planting area. Early sowing melons in the southern area, and leading to the most susceptible disease in the season from the melon expansion period to the positive plum rain in the mature period. Along with the increase of continuous cropping years, diseases are aggravated year by year, and the diseases can be aggravated by partial or repeated application of nitrogen fertilizer; melon continuous cropping, low topography or ponding after rain, high groundwater level, serious disease caused by phosphorus and potassium fertilizer deficiency and weak growth of field blocks, and rapid disease development; under the condition of cultivation in protected areas such as a greenhouse, a greenhouse and the like, close planting is difficult to occur due to poor ventilation and overhigh humidity.
Disclosure of Invention
The microbial agent has a good effect on preventing and treating gummy stem blight of muskmelon, has a good inhibition effect on gummy stem blight germs, obviously reduces the incidence rate of gummy stem blight of muskmelon, realizes the yield and income increase of muskmelon, has no problems of environmental pollution and the like compared with the traditional chemical agents, is nontoxic and residue-free, and is beneficial to environmental protection.
The invention adopts the following technical scheme to realize the purposes:
the microbial agent comprises, by weight, 10-15 parts of biological composite bacteria, 20-30 parts of diatomite, 40-50 parts of algal polysaccharide, 3-5 parts of zinc sulfate, 0.5-1 part of copper sulfate, 1-3 parts of sodium molybdate, 0.2-0.5 part of ferrous sulfate, 1-2 parts of potassium sulfate and 0.05-0.1 part of magnesium sulfate;
the biological compound bacteria in the microbial agent comprise non-hydroscopic streptomyces, bacillus amyloliquefaciens, trichoderma harzianum and pseudomonas fluorescens; the total viable bacteria concentration in the biological composite bacteria is more than or equal to 50 hundred million/g.
The preparation method of the biological composite bacteria comprises the following steps: taking waste edible fungi, adding activated non-hydroscopic streptomycete and trichoderma harzianum, and fermenting for 5-10 days in an environment with the temperature of 25-30 ℃ and the humidity of 50-70%; adding peanut meal, mixing, adding activated bacillus amyloliquefaciens and pseudomonas fluorescens, fermenting at 30-35deg.C under 50-70% humidity for 5-8 days, drying, pulverizing, and sieving to obtain biological composite bacteria.
Further, the adding amount of the non-water absorbing streptomycete is 0.4-0.6% of the weight of the waste edible fungi, the adding amount of the trichoderma harzianum is 0.1-0.3% of the weight of the waste edible fungi, the adding amount of the bacillus amyloliquefaciens is 0.3-0.5% of the weight of the waste edible fungi, and the adding amount of the pseudomonas fluorescens is 0.1-0.3% of the weight of the waste edible fungi.
Further, the weight ratio of the waste edible fungi to the peanut meal is 1 (0.5-0.8).
The invention provides a preparation method of the microbial agent, which comprises the following steps:
uniformly mixing biological composite bacteria, diatomite, zinc sulfate, copper sulfate, sodium molybdate, ferrous sulfate, potassium sulfate and magnesium sulfate to obtain a mixture; algal polysaccharide is prepared into algal polysaccharide solution according to a feed liquid ratio of 1 (2-3) kg/L; adding the mixture into algal polysaccharide solution, mixing well, drying, pulverizing, sieving to obtain microbial agent.
The invention provides a specific application of the microbial agent, namely the microbial agent can be used for preventing and treating melon gummy stem blight.
Furthermore, the microbial agent can be added into melon seedling raising matrix, and the addition amount is 0.2-0.5kg of the microbial agent per square seedling raising matrix.
Furthermore, the microbial agent can be added into a base fertilizer for basal application before planting melons, and the application amount is 1.5-3 kg/mu.
Furthermore, the microbial agent can be used for root irrigation during the growth period of melons, and the application amount is 1-2 kg/mu.
The invention has the following beneficial effects:
1. the invention effectively screens microbial flora to obtain that the combination of the streptomyces hygroscopicus, trichoderma harzianum, bacillus amyloliquefaciens and pseudomonas fluorescens can have better inhibition effect on the gummy stem blight fungus, wherein the streptomyces hygroscopicus and the trichoderma harzianum are main action flora, the bacillus amyloliquefaciens and the pseudomonas fluorescens are auxiliary flora, and the four flora are unavailable to form synergistic effect, so that the activity of the gummy stem blight fungus is inhibited.
2. In the preparation of the biological flora, the dominant flora of the streptomyces hygroscopicus and the trichoderma harzianum is formed firstly by adopting a batch fermentation mode, and the bacillus amyloliquefaciens and the pseudomonas fluorescens are added on the basis to form auxiliary flora, so that the inhibition effect of the biological compound bacteria obtained by compounding each flora according to the use amount on the gummy fungus is better than that of the traditional biological compound bacteria.
3. According to the invention, the edible fungus residues, peanut meal and other production wastes are used as fermentation matrixes, so that the recycling of resources is realized, meanwhile, certain fungi exist in the edible fungus residues, the fungi are also increased to a certain extent in the process of producing dominant fungus clusters by fermentation, the fungi play an auxiliary role in inhibiting gummy fungi, and when the edible fungus residues and the peanut meal are applied to melon seedlings, the edible fungus residues and the peanut meal can continuously provide nutrients for the fungus clusters, so that the advantages of the fungus clusters are ensured, and the fungus clusters are not lost due to the change of environment.
4. Compared with the traditional chemical agents, the microbial agent provided by the invention is nontoxic, pollution-free and residue-free, and ensures the food safety; meanwhile, the enhancement of the drug resistance of gummy stem blight fungi caused by long-term use of chemical agents is effectively avoided.
Detailed Description
The present invention is further illustrated below with reference to specific examples, which are to be construed as merely illustrative of the invention and not limiting of its scope, as various equivalent modifications to the invention will fall within the scope of the claims of the application after reading the invention.
Example 1
The preparation method of the biological composite bacteria comprises the following steps: taking 10kg of waste edible fungi, crushing, adding 0.4kg of activated streptomyces hygroscopicus and 0.3kg of trichoderma harzianum, uniformly mixing, and fermenting for 7 days in an environment with the temperature of 30 ℃ and the humidity of 70%, thus obtaining a primary fermentation product; adding 8kg of crushed peanut meal into the primary fermentation product, uniformly mixing, adding 0.3kg of activated bacillus amyloliquefaciens and 0.3kg of pseudomonas fluorescens, uniformly mixing, continuously fermenting for 3 days in an environment with the temperature of 30 ℃ and the humidity of 70%, obtaining a final fermentation product, drying, crushing and sieving the fermentation product, and thus obtaining the biological composite bacteria.
Uniformly mixing 15kg of biological composite bacteria, 20kg of diatomite, 5kg of zinc sulfate, 0.5kg of copper sulfate, 1kg of sodium molybdate, 0.5kg of ferrous sulfate, 2kg of potassium sulfate and 0.05kg of magnesium sulfate to obtain a mixture; taking 50kg of algal polysaccharide, adding 100L of water to prepare algal polysaccharide solution; adding the mixture into algal polysaccharide solution, mixing well, drying, pulverizing, sieving to obtain microbial agent.
Example 2
The preparation method of the biological composite bacteria comprises the following steps: taking 10kg of waste edible fungi, crushing, adding 0.6kg of activated streptomyces hygroscopicus and 0.1kg of trichoderma harzianum, uniformly mixing, and fermenting for 5 days in an environment with the temperature of 25 ℃ and the humidity of 50%, thus obtaining a primary fermentation product; adding 5kg of crushed peanut meal into the primary fermentation product, uniformly mixing, adding 0.5kg of activated bacillus amyloliquefaciens and 0.1kg of pseudomonas fluorescens, uniformly mixing, continuously fermenting for 5 days in an environment with the temperature of 35 ℃ and the humidity of 50%, obtaining a final fermentation product, drying, crushing and sieving the fermentation product, and thus obtaining the biological composite bacteria.
Uniformly mixing 10kg of biological composite bacteria, 30kg of diatomite, 3kg of zinc sulfate, 1kg of copper sulfate, 3kg of sodium molybdate, 0.2kg of ferrous sulfate, 1kg of potassium sulfate and 0.1kg of magnesium sulfate to obtain a mixture; taking 40kg of algal polysaccharide, and adding 120L of water to prepare algal polysaccharide solution; adding the mixture into algal polysaccharide solution, mixing well, drying, pulverizing, sieving to obtain microbial agent.
Example 3
The preparation method of the biological composite bacteria comprises the following steps: taking 10kg of waste edible fungi, crushing, adding 0.5kg of activated streptomyces hygroscopicus and 0.2kg of trichoderma harzianum, uniformly mixing, and fermenting for 6 days in an environment with the temperature of 30 ℃ and the humidity of 50%, thus obtaining a primary fermentation product; adding 6kg of crushed peanut meal into the primary fermentation product, uniformly mixing, adding 0.4kg of activated bacillus amyloliquefaciens and 0.2kg of pseudomonas fluorescens, uniformly mixing, continuously fermenting for 4 days in an environment with the temperature of 25 ℃ and the humidity of 50%, obtaining a final fermentation product, drying, crushing and sieving the fermentation product, and thus obtaining the biological composite bacteria.
Uniformly mixing 13kg of biological composite bacteria, 15kg of diatomite, 4kg of zinc sulfate, 0.8kg of copper sulfate, 2kg of sodium molybdate, 0.4kg of ferrous sulfate, 1kg of potassium sulfate and 0.07kg of magnesium sulfate to obtain a mixture; taking 50kg of algal polysaccharide, adding 100L of water to prepare algal polysaccharide solution; adding the mixture into algal polysaccharide solution, mixing well, drying, pulverizing, sieving to obtain microbial agent.
Comparative example 1
The preparation method of the biological composite bacteria comprises the following steps: taking 10kg of waste edible fungi, crushing, adding activated 0.5kg of non-hydroscopic streptomycete, 0.2kg of trichoderma harzianum, 0.4kg of bacillus amyloliquefaciens and 0.2kg of pseudomonas fluorescens, uniformly mixing, continuously fermenting for 4 days in an environment with the temperature of 25 ℃ and the humidity of 50%, obtaining a fermented product, drying, crushing and sieving the fermented product, thus obtaining the biological composite fungus.
Uniformly mixing 13kg of biological composite bacteria, 15kg of diatomite, 4kg of zinc sulfate, 0.8kg of copper sulfate, 2kg of sodium molybdate, 0.4kg of ferrous sulfate, 1kg of potassium sulfate and 0.07kg of magnesium sulfate to obtain a mixture; taking 50kg of algal polysaccharide, adding 100L of water to prepare algal polysaccharide solution; adding the mixture into algal polysaccharide solution, mixing well, drying, pulverizing, sieving to obtain microbial agent.
Comparative example 2
The preparation method of the biological composite bacteria comprises the following steps: taking 10kg of peanut meal, crushing, adding activated 0.5kg of non-hydroscopic streptomycete and 0.2kg of trichoderma harzianum, uniformly mixing, fermenting for 6 days in an environment with the temperature of 30 ℃ and the humidity of 50%, obtaining a fermented product, drying, crushing and sieving the fermented product, and obtaining the biological composite bacteria.
Uniformly mixing 13kg of biological composite bacteria, 15kg of diatomite, 4kg of zinc sulfate, 0.8kg of copper sulfate, 2kg of sodium molybdate, 0.4kg of ferrous sulfate, 1kg of potassium sulfate and 0.07kg of magnesium sulfate to obtain a mixture; taking 50kg of algal polysaccharide, adding 100L of water to prepare algal polysaccharide solution; adding the mixture into algal polysaccharide solution, mixing well, drying, pulverizing, sieving to obtain microbial agent.
Comparative example 3
The preparation method of the biological composite bacteria comprises the following steps: taking 10kg of waste edible fungi, crushing, adding 0.4kg of activated bacillus amyloliquefaciens and 0.2kg of pseudomonas fluorescens, uniformly mixing, and fermenting for 6 days in an environment with the temperature of 30 ℃ and the humidity of 50% to obtain a first fermentation product; adding 6kg of crushed peanut meal into the primary fermentation product, uniformly mixing, adding 0.5kg of activated streptomyces hygroscopicus and 0.2kg of trichoderma harzianum, uniformly mixing, continuously fermenting for 4 days in an environment with the temperature of 25 ℃ and the humidity of 50%, obtaining a final fermentation product, drying, crushing and sieving the fermentation product, and obtaining the biological composite bacteria.
Uniformly mixing 13kg of biological composite bacteria, 15kg of diatomite, 4kg of zinc sulfate, 0.8kg of copper sulfate, 2kg of sodium molybdate, 0.4kg of ferrous sulfate, 1kg of potassium sulfate and 0.07kg of magnesium sulfate to obtain a mixture; taking 50kg of algal polysaccharide, adding 100L of water to prepare algal polysaccharide solution; adding the mixture into algal polysaccharide solution, mixing well, drying, pulverizing, sieving to obtain microbial agent.
Performance testing
1. Plate antagonism of the microbial agent of the invention to gummy stem blight germ
Taking 3g of biological composite bacteria prepared according to the methods of examples 1-3 and comparative examples 1-3, adding a proper amount of water, stirring uniformly, and filtering to obtain filtrate for later use.
Inoculating gummy stem blight fungus to the central position on the PDA plate, inoculating the obtained filtrate with sterile droppers at two opposite positions equidistant from the center, inoculating sterilized water at two opposite corners as control, inverting the PDA plate, culturing in a incubator at 25deg.C, and observing the growth of gummy stem blight fungus every two days.
The bacterial plaque radius of the biological compound bacteria of the inoculation examples 1-3 is above 1.0cm, the distance from the edge of the bacterial plaque to the edge of the bacterial plaque of the gummy stem fungus is above 1.3cm, and the bacterial plaque radius of the gummy stem fungus at the control position is about 3.3 cm. The radius of bacterial plaque of the biological compound bacteria inoculated in the comparative example 1 is about 0.8cm, the distance between the bacterial plaque and the edge of bacterial plaque of the gummy stem fungus is about 0.9cm, and the radius of bacterial plaque of the gummy stem fungus at the control position is about 3.5 cm; the radius of bacterial plaque inoculated with the biological compound bacteria of comparative example 2 is about 0.5cm, the distance between the bacterial plaque radius and the edge of bacterial plaque of gummy stem blight fungus is about 0.5cm, and the bacterial plaque of gummy stem blight fungus at the control position is about 3.8 cm; the radius of the bacterial plaque of the biological composite bacteria inoculated in comparative example 3 was about 0.7cm, the distance from the edge of the bacterial plaque of the gummy stem fungus was about 0.4cm, and the radius of the bacterial plaque of the gummy stem fungus at the control place was about 3.6 cm.
The data show that the biological composite bacteria prepared by the invention have better effect of inhibiting the activity of gummy stem blight fungi.
In the biological composite flora prepared by the invention, the inhibition effect on the gummy fungus is mainly that the gummy fungus is streptomyces hygroscopicus and trichoderma harzianum, the bacillus amyloliquefaciens and the pseudomonas fluorescens are auxiliary effects, and certain fungi exist in edible fungus residues and can also play an auxiliary effect to a certain extent, and various flora are mutually matched to realize the efficient inhibition on the gummy fungus.
The biological compound strains in the comparative example 1 are fermented together, and bacillus amyloliquefaciens and pseudomonas fluorescens are produced by fermentation in the comparative example 3, and the two fermentation modes lead to that streptomyces hygroscopicus and trichoderma harzianum do not form dominant bacterial groups, so that the inhibition effect of the biological compound strains on gummy fungi is reduced; in comparative example 2, bacillus amyloliquefaciens and pseudomonas fluorescens are absent, and edible fungus residues are not used as fermentation matrixes, so that the inhibition effect of the biological composite bacteria on gummy fungus is obviously weakened.
2. The microbial agent of the invention has disease prevention effect on melon gummy stem blight
20g of the biological composite bacteria prepared according to the methods of examples 1-3 and comparative examples 1-3 is dissolved in 1500mL of water to obtain bacterial liquid for later use;
transplanting melon seedlings with 5 leaf periods and consistent growth vigor into pots, transplanting one melon seedling in each pot, and transplanting 400 melon seedlings in total, wherein each group is divided into 8 groups of 50 melon seedlings, namely a blank group, a control group, an experiment 1 group, an experiment 2 group, an experiment 3 group, an experiment 4 group, an experiment 5 group and an experiment 6 group.
On the day of transplanting, 1-6 groups of experiments respectively use biological compound bacteria solution to irrigate root of melon seedlings, and each melon seedling is irrigated with 20mL; the root of each melon seedling in the blank group and the control group is irrigated with 20mL of clear water;
the next day of transplanting, 20mL of gummy stem blight fungus spore suspension is respectively irrigated to the seedlings of the control group and the experiment 1-6 groups, and the spore concentration is 10 multiplied by 10 6 individual/mL; and 20mL of clear water is irrigated into roots of each melon seedling in the blank group.
In a greenhouse environment at 25 ℃, observing the conditions of melon seedlings in each group every day, starting counting the disease conditions of the melon seedlings in each group after the disease conditions appear in a control group, and stopping observing after the disease rate of the control group reaches more than 90%.
Table 1 number and incidence of melon seedlings of each group
| Group of | Bacterial liquid | Number of affected plants | Incidence of disease |
| Blank group | — | 10 | 20 |
| Control group | — | 45 | 90 |
| Experiment 1 group | Example 1 | 8 | 16 |
| Experiment 2 group | Example 2 | 9 | 18 |
| Experiment 3 group | Example 3 | 5 | 10 |
| Experiment 4 group | Comparative example 1 | 19 | 38 |
| Experiment 5 group | Comparative example 2 | 29 | 58 |
| Experiment 6 group | Comparative example 3 | 23 | 46 |
As can be seen from the data in Table 1, the incidence of melon seedlings (experiments 1, 2 and 3) irrigated with the biological compound bacteria prepared by the invention is below 20%, and the incidence of melon seedlings with the biological compound bacteria obtained in comparative examples 1-3 is higher and above 35%; the blank melon seedlings are in the same greenhouse environment as other melon seedlings, are easy to be infected by gummy fungus spores, and cause morbidity, and the morbidity rate is about 20% and is higher than that of experiments 1-3. The biological composite bacteria prepared by the invention has better inhibition effect on the gummy stem blight fungi, and can obviously reduce the incidence rate of the gummy stem blight.
3. Field experiments test the prevention and treatment effect of the microbial agent of the invention on melon gummy stem blight
The field experiment is carried out on a melon planting base of Lanling, shandong province, the variety is Boyang 99, and the area of each treatment area is 40m 2 100 melons are planted in each area, wherein the plant spacing is 35-40cm; a guard row 1.5m wide is established between adjacent processing regions. And setting 7 treatment areas which are a control area, an experiment 1 area, an experiment 2 area, an experiment 3 area, an experiment 4 area, an experiment 5 area and an experiment 6 area respectively, and performing field management on each area according to a conventional cultivation management technology.
Root irrigation treatment is carried out on each experimental area on the day of transplanting melon seedlings, 10 th day, 20 th day and 30 th day by using corresponding microbial agents, and 100mL (about 1.2g containing the microbial agents) of each melon seedling is irrigated; the control area was rooted with 100mL of clear water.
After the onset of the control group, the onset of each experimental area was observed.
Table 2 number of and incidence of melon seedlings in each group
| Group of | Bacterial liquid | Number of affected plants | Incidence of disease |
| Control group | — | 16 | 16％ |
| Experiment 1 group | Example 1 | 4 | 4％ |
| Experiment 2 group | Example 2 | 1 | 1％ |
| Experiment 3 group | Example 3 | 2 | 2％ |
| Experiment 4 group | Comparative example 1 | 6 | 6％ |
| Experiment 5 group | Comparative example 2 | 10 | 10％ |
| Experiment 6 group | Comparative example 3 | 7 | 7％ |
As can be seen from the data in Table 2, the microbial inoculum prepared in examples 1-3 of the invention is used for root-filling melon seedlings, the incidence rate of gummy stem blight is lower than 5% during the growth period of melon seedlings; the incidence of melon seedlings using the microbial agents obtained in comparative examples 1 to 3 was slightly higher, wherein the incidence of melon seedlings using the microbial agents obtained in comparative example 2 was 10% higher than others.
Claims (9)
1. The microbial agent is characterized by comprising, by weight, 10-15 parts of biological composite bacteria, 20-30 parts of diatomite, 40-50 parts of algal polysaccharide, 3-5 parts of zinc sulfate, 0.5-1 part of copper sulfate, 1-3 parts of sodium molybdate, 0.2-0.5 part of ferrous sulfate, 1-2 parts of potassium sulfate and 0.05-0.1 part of magnesium sulfate; wherein the biological composite bacteria comprise non-hydroscopic streptomyces, bacillus amyloliquefaciens, trichoderma harzianum and pseudomonas fluorescens; the total viable bacteria concentration in the biological composite bacteria is more than or equal to 50 hundred million/g.
2. The microbial agent of claim 1, wherein the preparation method of the biological composite bacteria comprises the following steps: taking waste edible fungi, adding activated non-hydroscopic streptomycete and trichoderma harzianum, and fermenting for 5-7 days in an environment with the temperature of 25-30 ℃ and the humidity of 50-70%; adding peanut meal, mixing, adding activated bacillus amyloliquefaciens and pseudomonas fluorescens, fermenting at 30-35deg.C under 50-70% humidity for 3-5 days, drying, pulverizing, and sieving to obtain the final product.
3. The microbial agent according to claim 2, wherein in the preparation method of the biological composite bacteria, the adding amount of the non-hydroscopic streptomycete is 0.4-0.6% of the weight of the waste edible fungi, the adding amount of the trichoderma harzianum is 0.1-0.3% of the weight of the waste edible fungi, the adding amount of the bacillus amyloliquefaciens is 0.3-0.5% of the weight of the waste edible fungi, and the adding amount of the fluorescent pseudomonas is 0.1-0.3% of the weight of the waste edible fungi.
4. The microbial agent according to claim 2, wherein in the preparation method of the biological composite bacteria, the weight ratio of the waste edible fungi to the peanut meal is 1 (0.5-0.8).
5. A process for the preparation of a microbial agent as claimed in any one of claims 1 to 4, comprising the steps of: uniformly mixing biological composite bacteria, diatomite, zinc sulfate, copper sulfate, sodium molybdate, ferrous sulfate, potassium sulfate and magnesium sulfate to obtain a mixture; algal polysaccharide is prepared into algal polysaccharide solution according to a feed liquid ratio of 1 (2-3) kg/L; adding the mixture into algal polysaccharide solution, mixing well, drying, pulverizing, sieving to obtain microbial agent.
6. Use of a microbial agent according to any one of claims 1 to 4 for the control of melon gummy stem blight.
7. The use according to claim 6, wherein the microbial agent is added to melon seedling substrates in an amount of 0.2-0.5kg per seedling substrate.
8. The use according to claim 6, wherein the microbial inoculum is applied singly or added to the base fertilizer before the melon is planted, and the application amount is 1.5-3 kg/mu.
9. The use according to claim 6, wherein the microbial inoculum is root irrigated during melon growth at an application rate of 1-2 kg/mu.
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| CN117778276A (en) * | 2024-02-23 | 2024-03-29 | 四川省农业农村厅植物保护站 | Biocontrol microbial inoculum for preventing and treating soil-borne diseases of ginger, preparation method and cultivation method |
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| CN104247717A (en) * | 2013-06-25 | 2014-12-31 | 中国科学院沈阳应用生态研究所 | Microbial inoculant inhibiting diseases and insect pests of plants, and preparation method and application thereof |
| CN107245461A (en) * | 2017-05-31 | 2017-10-13 | 北方民族大学 | A kind of bacillus amyloliquefaciens B6 and its application |
| CN109055265A (en) * | 2018-08-16 | 2018-12-21 | 南京农业大学 | A kind of biocontrol microorganisms and its application in terms of crops blight dis-ease prevention and treatment |
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| CN102329755A (en) * | 2011-09-28 | 2012-01-25 | 中国热带农业科学院海口实验站 | Nonabsorbent streptomyces ahygroscopicus strain BWL58 for preventing and treating banana vascular wilt and application thereof |
| CN104247717A (en) * | 2013-06-25 | 2014-12-31 | 中国科学院沈阳应用生态研究所 | Microbial inoculant inhibiting diseases and insect pests of plants, and preparation method and application thereof |
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| Publication number | Priority date | Publication date | Assignee | Title |
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| CN117778276A (en) * | 2024-02-23 | 2024-03-29 | 四川省农业农村厅植物保护站 | Biocontrol microbial inoculum for preventing and treating soil-borne diseases of ginger, preparation method and cultivation method |
| CN117778276B (en) * | 2024-02-23 | 2024-05-10 | 四川省农业农村厅植物保护站 | Biocontrol microbial inoculum for preventing and treating soil-borne diseases of ginger, preparation method and cultivation method |
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