CN114395548A - Compound microbial preparation for preventing and treating root-knot nematode and application thereof - Google Patents
Compound microbial preparation for preventing and treating root-knot nematode and application thereof Download PDFInfo
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- 239000000843 powder Substances 0.000 claims abstract description 197
- 241000187395 Streptomyces microflavus Species 0.000 claims abstract description 54
- 239000004575 stone Substances 0.000 claims abstract description 44
- 241000193830 Bacillus <bacterium> Species 0.000 claims abstract description 27
- 241000191025 Rhodobacter Species 0.000 claims abstract description 25
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- 239000003795 chemical substances by application Substances 0.000 claims abstract description 6
- 238000000855 fermentation Methods 0.000 claims description 51
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- 241001495152 Rhodovulum sulfidophilum Species 0.000 claims description 41
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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
- C12N11/00—Carrier-bound or immobilised enzymes; Carrier-bound or immobilised microbial cells; Preparation thereof
- C12N11/02—Enzymes or microbial cells immobilised on or in an organic carrier
-
- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01G—HORTICULTURE; CULTIVATION OF VEGETABLES, FLOWERS, RICE, FRUIT, VINES, HOPS OR SEAWEED; FORESTRY; WATERING
- A01G13/00—Protecting plants
-
- 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
-
- 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
- A01N63/28—Streptomyces
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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
- C12N1/04—Preserving or maintaining viable microorganisms
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
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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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- 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
- C12N11/00—Carrier-bound or immobilised enzymes; Carrier-bound or immobilised microbial cells; Preparation thereof
- C12N11/14—Enzymes or microbial cells immobilised on or in an inorganic carrier
Abstract
The invention discloses a compound microbial preparation for preventing and treating root-knot nematode, which consists of waxy bacillus powder, sulphur-philic rhodobacter xylinum powder, streptomyces microflavus powder and a carrier; the carrier consists of crop straw powder and modified medical stone powder; the effective strains comprise waxy bacillus powder, sulphur-philic rhodobacter parvum powder and streptomyces microflavus powder; the three strains are used in a compounding way, have synergistic effect and are mutually synergistic; solves the problems of poor prevention and treatment effect and poor long-term stability of single microbial agent on the root-knot nematode. The microbial preparation is used for preventing and treating root-knot nematodes in a biological prevention and treatment manner, is green and environment-friendly, has a good effect on preventing and treating the root-knot nematodes, and can promote the growth of crops by microbial secretions to achieve the effect of increasing yield.
Description
Technical Field
The invention belongs to the field of microbial preparations, and particularly relates to a compound microbial preparation for preventing and treating root-knot nematodes and an application thereof.
Background
Root-knot nematodes, which are parasitic plant nematodes, mainly damage the roots of plants and mainly appear as root nodules formed on the roots of the plants, causing poor growth of the roots of the plants, and after the roots of the plants are attacked by the nematodes, the roots of the plants affect the absorption of nutrients and water in soil, causing wilting of the overground parts of the plants, slow and short growth, low yield and even death of the plants; the yield of crops is reduced, and serious economic loss is brought to farmers;
at present, chemical pesticides are mostly adopted in a method for preventing and treating root-knot nematodes, the pesticide resistance of the root-knot nematodes can be caused by long-term use of the pesticides, the pesticide effect is weakened, meanwhile, part of the chemical pesticides can cause pollution to farmland soil, and the existing method for preventing and treating insect pests by adopting a microbial agent is a method for expelling the insect pests or harmful bacteria by utilizing substances capable of resisting the insect pests or the harmful bacteria generated in the metabolic process of the microbial agent. The existing single-kind microbial agent has limited control effect on the root-knot nematode; the effect of improving soil can not be achieved, the continuous stability of the prevention and control effect of the root-knot nematode is poor, and the plants are repeatedly infested by the root-knot nematode due to continuous cropping cultivation.
Disclosure of Invention
Aiming at the defects of the prior art, the invention provides a compound microbial preparation for preventing and treating root-knot nematodes, wherein the effective strains of the compound microbial preparation comprise bacillus cereus powder, rhodobacter sulfidophilus powder and streptomyces microflavus powder; the three strains are used in a compounding way, have synergistic effect and are mutually synergistic; solves the problems of poor prevention and treatment effect and poor long-term stability of single microbial agent on the root-knot nematode. The microbial preparation is used for preventing and treating root-knot nematodes in a biological prevention and treatment manner, is green and environment-friendly, has a good effect on preventing and treating the root-knot nematodes, and can promote the growth of crops by microbial secretions to achieve the effect of increasing yield.
In order to solve the technical problems, the invention adopts the following technical scheme:
a compound microbial preparation for preventing and treating root-knot nematode is composed of waxy bacillus powder, sulphur-philic rhodobacter xylinum powder, streptomyces microflavus powder and a carrier; the carrier consists of crop straw powder and modified medical stone powder; the preparation method specifically comprises the following steps:
(1) preparing microbial powder:
respectively carrying out liquid fermentation culture on bacillus cereus, rhodobacter sulfidophilus and streptomyces microflavus to obtain fermentation liquor, adding a protective agent A into the bacillus cereus fermentation liquor, carrying out emulsification treatment, and carrying out spray drying to obtain bacillus cereus powder; adding a protective agent A into the rhodobacter sulfidophilus fermentation liquor, emulsifying, and then carrying out spray drying to obtain rhodobacter sulfidophilus powder; adding a protective agent B into the streptomyces microflavus fermentation liquor, emulsifying, and then carrying out spray drying to obtain rhodobacter sulfidophilus powder;
(2) preparation of Carrier Dispersion
Uniformly mixing crop straw powder and modified medical stone powder, and then adding water to prepare a mixed carrier dispersion liquid with the mass percentage of 50-55%;
(3) preparation of composite microbial preparation
Mixing the waxy bacillus powder, the sulphur-philic rhodobacter parvum powder and the streptomyces microflavus powder in the step (1) to obtain mixed bacteria powder; and (3) adding the mixed bacterial powder into the mixed carrier dispersion liquid in the step (2), uniformly stirring, and drying to obtain the composite microbial preparation.
Preferably, the protective agent A in the step (1) consists of modified gelatin and cyclodextrin, and the weight ratio of the modified gelatin to the cyclodextrin is 1: 1; the protective agent B is cyclodextrin;
preferably, the addition amount of the protective agent A accounts for 20 percent of the weight of the bacillus cereus fermentation liquor and the rhodobacter sulfidophilus fermentation liquor; the addition amount of the protective agent B accounts for 20 percent of the weight of the streptomyces microflavus fermentation liquor;
preferably, the weight ratio of the waxy bacillus powder, the rhodobacter sulfidophilus powder and the streptomyces microflavus powder in the step (2) is 5-7: 3-5: 1-3;
preferably, the weight ratio of the waxy bacillus powder, the sulfur-philic rhodobacter xylinum powder and the streptomyces microflavus powder in the step (2) is 6:4: 2;
preferably, the weight ratio of the crop straw powder to the modified medical stone powder in the step (2) is 2: 1;
preferably, the weight ratio of the adding amount of the mixed bacterial powder in the step (3) to the mixed carrier dispersion liquid is 6: 25;
preferably, the modified medical stone is sulfhydryl modified medical stone, and the preparation method comprises the following steps:
1) sequentially carrying out acid leaching on medical stone, washing with water until washing liquid is neutral, drying, and grinding to obtain medical stone powder;
2) dispersing medical stone in an ethanol solution, and dropwise adding a sulfhydryl modifier into the ethanol solution; heating and stirring at 50-60 deg.C for 2-3 hr, filtering, washing, drying, and grinding to obtain sulfhydryl modified Maifanitum;
furthermore, the mercapto modifier is mercaptopropyltrimethoxysilane;
further, the concentration of the ethanol solution is 50%; the weight volume ratio of the medical stone to the ethanol solution is 50 g/L; the weight ratio of the addition amount of the sulfhydryl modifier to the medical stone is 1: 5;
the modified gelatin is gelatin treated by gelatinase, and specifically comprises adding water into gelatin, stirring for swelling, stirring at 90 deg.C for dissolving completely, cooling to room temperature, adding gelatinase, stirring at room temperature for reaction, and inactivating enzyme to obtain modified gelatin;
furthermore, the effective viable count in the composite microbial preparation is more than or equal to 0.2 hundred million/gram;
the application method of the compound microbial preparation for preventing and treating the root-knot nematode can select any one of drip irrigation, flushing application, broadcast application and root irrigation, and the using amount is 10-20 kg/mu.
Advantageous effects
In the microbial preparation prepared by the invention, bacillus cereus, rhodobacter sulfidophilum and streptomyces microflavus have synergistic effect in the prevention and treatment of root-knot nematodes, and form a dominant flora in soil, so that the dominant flora can parasitize in eggs of the root-knot nematodes, cyst nematodes, stem nematodes, nematodes and the like and can infect larvae and females; on the other hand, each strain can secrete and synthesize various functional enzymes including chitinase, cell lyase, glucanase, serine protease and other metabolites such as abundant indoleacetic acid derivatives; not only achieves good effect of preventing and controlling root-knot nematode; the metabolite can also promote seed germination, plant root system and plant growth.
The protective agent added into the compound microbial preparation has the functions of promoting the growth of flora and stimulating the secretion of flora; the growth activity of the flora and the survival period of the flora in the soil can be effectively improved, and the long-term function of the flora is promoted;
the composite microbial preparation also contains crop straw powder and sulfhydryl modified medical stone powder, so that a proper living space is provided for the microbes; and can also improve the physicochemical property of soil, promote the growth of plants and reduce the risk of insect attack on plants.
The synergistic effect of the bacillus cereus, the rhodobacter sulfidophilum and the streptomyces microflavus is that when the weight ratio of the bacillus cereus, the rhodobacter sulfidophilum and the streptomyces microflavus is 6:4:2, the prevention and treatment effect on root-knot nematode is the best, the growth of cucumbers can be promoted, and the yield of the cucumbers is remarkably improved.
Detailed Description
The technical solution of the present invention is described in detail with reference to the specific embodiments, but it should be emphasized that the present invention is not limited to the specific embodiments; the microbial species used in the examples below can be isolated by themselves or obtained by purchase; the starting materials described in the examples below are, without specific indication, either prepared by conventional methods or commercially available.
Example 1
Preparing sulfhydryl modified medical stone:
1) sequentially carrying out acid soaking on medical stone, washing with water until washing liquid is neutral, drying, and grinding to obtain medical stone powder with the particle size of 200-300 meshes;
2) dispersing medical stone powder in 50% ethanol solution (volume fraction), and adding mercaptopropyl trimethoxysilane dropwise into the solution; heating and stirring at 50-60 deg.C for 2-3 hr, filtering, washing, drying, and grinding to obtain sulfhydryl modified Maifanitum; wherein the weight ratio of the mercaptopropyl trimethoxy silane to the medical stone powder is 1: 5; the addition amount of the medical stone powder accounts for 50g/L of the weight volume ratio of the ethanol solution;
preparation of modified gelatin
Adding water into gelatin, stirring for swelling, stirring at 90 deg.C until completely dissolved, cooling to room temperature, adding gelatinase, stirring at room temperature for reaction, and inactivating enzyme to obtain modified gelatin;
example 2
A compound microbial preparation for preventing and treating root-knot nematode is composed of waxy bacillus powder, sulphur-philic rhodobacter xylinum powder, streptomyces microflavus powder and a carrier; the carrier consists of crop straw powder and modified medical stone powder; the preparation method specifically comprises the following steps:
(1) preparing microbial powder:
respectively carrying out liquid fermentation culture on bacillus cereus, rhodobacter sulfidophilus and streptomyces microflavus to obtain fermentation liquor, adding a mixture of 2kg of modified gelatin and cyclodextrin which are mixed by equal weight into 10kg of bacillus cereus fermentation liquor, emulsifying, and carrying out spray drying to obtain bacillus cereus powder; adding 2kg of a mixture of modified gelatin and cyclodextrin which are mixed by equal weight into 10kg of rhodobacter sulfidophilus fermentation liquor, emulsifying, and performing spray drying to obtain rhodobacter sulfidophilus powder; adding 2kg of cyclodextrin into 10kg of streptomyces microflavus fermentation liquor for emulsification, and performing spray drying to obtain rhodobacter sulfidophilus powder;
(2) preparation of Carrier Dispersion
Mixing 17kg of crop straw powder and 8.5kg of modified medical stone powder uniformly, and adding water to 50kg of the mixture to prepare mixed carrier dispersion liquid;
(3) preparation of composite microbial preparation
Mixing 5kg of waxy bacillus powder, 5kg of sulphur-philic rhodobacter parvum powder and 2kg of streptomyces microflavus powder in the step (1) to obtain mixed bacteria powder; and (3) adding the mixed bacterial powder into the mixed carrier dispersion liquid in the step (2), uniformly stirring, and drying to obtain the composite microbial preparation.
Example 3
A compound microbial preparation for preventing and treating root-knot nematode is composed of waxy bacillus powder, sulphur-philic rhodobacter xylinum powder, streptomyces microflavus powder and a carrier; the carrier consists of crop straw powder and modified medical stone powder; the preparation method specifically comprises the following steps:
(1) preparing microbial powder:
respectively carrying out liquid fermentation culture on bacillus cereus, rhodobacter sulfidophilus and streptomyces microflavus to obtain fermentation liquor, adding a mixture of 2kg of modified gelatin and cyclodextrin which are mixed by equal weight into 10kg of bacillus cereus fermentation liquor, emulsifying, and carrying out spray drying to obtain bacillus cereus powder; adding 2kg of a mixture of modified gelatin and cyclodextrin which are mixed by equal weight into 10kg of rhodobacter sulfidophilus fermentation liquor, emulsifying, and performing spray drying to obtain rhodobacter sulfidophilus powder; adding 2kg of cyclodextrin into 10kg of streptomyces microflavus fermentation liquor for emulsification, and performing spray drying to obtain rhodobacter sulfidophilus powder;
(2) preparation of Carrier Dispersion
Mixing 17kg of crop straw powder and 8.5kg of modified medical stone powder uniformly, and adding water to 50kg of the mixture to prepare mixed carrier dispersion liquid;
(3) preparation of composite microbial preparation
Mixing 5kg of waxy bacillus powder, 4kg of sulphur-philic rhodobacter parvum powder and 3kg of streptomyces microflavus powder in the step (1) to obtain mixed bacteria powder; and (3) adding the mixed bacterial powder into the mixed carrier dispersion liquid in the step (2), uniformly stirring, and drying to obtain the composite microbial preparation.
Example 4
A compound microbial preparation for preventing and treating root-knot nematode is composed of waxy bacillus powder, sulphur-philic rhodobacter xylinum powder, streptomyces microflavus powder and a carrier; the carrier consists of crop straw powder and modified medical stone powder; the preparation method specifically comprises the following steps:
(1) preparing microbial powder:
respectively carrying out liquid fermentation culture on bacillus cereus, rhodobacter sulfidophilus and streptomyces microflavus to obtain fermentation liquor, adding a mixture of 2kg of modified gelatin and cyclodextrin which are mixed by equal weight into 10kg of bacillus cereus fermentation liquor, emulsifying, and carrying out spray drying to obtain bacillus cereus powder; adding 2kg of a mixture of modified gelatin and cyclodextrin which are mixed by equal weight into 10kg of rhodobacter sulfidophilus fermentation liquor, emulsifying, and performing spray drying to obtain rhodobacter sulfidophilus powder; adding 2kg of cyclodextrin into 10kg of streptomyces microflavus fermentation liquor for emulsification, and performing spray drying to obtain rhodobacter sulfidophilus powder;
(2) preparation of Carrier Dispersion
Mixing 17kg of crop straw powder and 8.5kg of modified medical stone powder uniformly, and adding water to 50kg of the mixture to prepare mixed carrier dispersion liquid;
(3) preparation of composite microbial preparation
Mixing 6kg of waxy bacillus powder, 3kg of sulphur-philic rhodobacter parvum powder and 3kg of streptomyces microflavus powder in the step (1) to obtain mixed bacteria powder; and (3) adding the mixed bacterial powder into the mixed carrier dispersion liquid in the step (2), uniformly stirring, and drying to obtain the composite microbial preparation.
Example 5
A compound microbial preparation for preventing and treating root-knot nematode is composed of waxy bacillus powder, sulphur-philic rhodobacter xylinum powder, streptomyces microflavus powder and a carrier; the carrier consists of crop straw powder and modified medical stone powder; the preparation method specifically comprises the following steps:
(1) preparing microbial powder:
respectively carrying out liquid fermentation culture on bacillus cereus, rhodobacter sulfidophilus and streptomyces microflavus to obtain fermentation liquor, adding a mixture of 2kg of modified gelatin and cyclodextrin which are mixed by equal weight into 10kg of bacillus cereus fermentation liquor, emulsifying, and carrying out spray drying to obtain bacillus cereus powder; adding 2kg of a mixture of modified gelatin and cyclodextrin which are mixed by equal weight into 10kg of rhodobacter sulfidophilus fermentation liquor, emulsifying, and performing spray drying to obtain rhodobacter sulfidophilus powder; adding 2kg of cyclodextrin into 10kg of streptomyces microflavus fermentation liquor for emulsification, and performing spray drying to obtain rhodobacter sulfidophilus powder;
(2) preparation of Carrier Dispersion
Mixing 17kg of crop straw powder and 8.5kg of modified medical stone powder uniformly, and adding water to 50kg of the mixture to prepare mixed carrier dispersion liquid;
(3) preparation of composite microbial preparation
Mixing 6kg of waxy bacillus powder, 4kg of sulphur-philic rhodobacter parvum powder and 2kg of streptomyces microflavus powder in the step (1) to obtain mixed bacteria powder; and (3) adding the mixed bacterial powder into the mixed carrier dispersion liquid in the step (2), uniformly stirring, and drying to obtain the composite microbial preparation.
Example 6
A compound microbial preparation for preventing and treating root-knot nematode is composed of waxy bacillus powder, sulphur-philic rhodobacter xylinum powder, streptomyces microflavus powder and a carrier; the carrier consists of crop straw powder and modified medical stone powder; the preparation method specifically comprises the following steps:
(1) preparing microbial powder:
respectively carrying out liquid fermentation culture on bacillus cereus, rhodobacter sulfidophilus and streptomyces microflavus to obtain fermentation liquor, adding a mixture of 2kg of modified gelatin and cyclodextrin which are mixed by equal weight into 10kg of bacillus cereus fermentation liquor, emulsifying, and carrying out spray drying to obtain bacillus cereus powder; adding 2kg of a mixture of modified gelatin and cyclodextrin which are mixed by equal weight into 10kg of rhodobacter sulfidophilus fermentation liquor, emulsifying, and performing spray drying to obtain rhodobacter sulfidophilus powder; adding 2kg of cyclodextrin into 10kg of streptomyces microflavus fermentation liquor for emulsification, and performing spray drying to obtain rhodobacter sulfidophilus powder;
(2) preparation of Carrier Dispersion
Mixing 17kg of crop straw powder and 8.5kg of modified medical stone powder uniformly, and adding water to 50kg of the mixture to prepare mixed carrier dispersion liquid;
(3) preparation of composite microbial preparation
Mixing 6kg of waxy bacillus powder, 6kg of sulphur-philic rhodobacter parvum powder and 1kg of streptomyces microflavus powder in the step (1) to obtain mixed bacteria powder; and (3) adding the mixed bacterial powder into the mixed carrier dispersion liquid in the step (2), uniformly stirring, and drying to obtain the composite microbial preparation.
Example 7
(1) Preparing microbial powder:
respectively carrying out liquid fermentation culture on bacillus cereus, rhodobacter sulfidophilus and streptomyces microflavus to obtain fermentation liquor, adding a mixture of 2kg of modified gelatin and cyclodextrin which are mixed by equal weight into 10kg of bacillus cereus fermentation liquor, emulsifying, and carrying out spray drying to obtain bacillus cereus powder; adding 2kg of a mixture of modified gelatin and cyclodextrin which are mixed by equal weight into 10kg of rhodobacter sulfidophilus fermentation liquor, emulsifying, and performing spray drying to obtain rhodobacter sulfidophilus powder; adding 2kg of cyclodextrin into 10kg of streptomyces microflavus fermentation liquor for emulsification, and performing spray drying to obtain rhodobacter sulfidophilus powder;
(2) preparation of Carrier Dispersion
Mixing 17kg of crop straw powder and 8.5kg of modified medical stone powder uniformly, and adding water to 50kg of the mixture to prepare mixed carrier dispersion liquid;
(3) preparation of composite microbial preparation
Mixing 7kg of waxy bacillus powder, 4kg of sulphur-philic rhodobacter parvum powder and 1kg of streptomyces microflavus powder in the step (1) to obtain mixed bacteria powder; and (3) adding the mixed bacterial powder into the mixed carrier dispersion liquid in the step (2), uniformly stirring, and drying to obtain the composite microbial preparation.
Example 8
A compound microbial preparation for preventing and treating root-knot nematode is composed of waxy bacillus powder, sulphur-philic rhodobacter xylinum powder, streptomyces microflavus powder and a carrier; the carrier consists of crop straw powder and modified medical stone powder; the preparation method specifically comprises the following steps:
(1) preparing microbial powder:
respectively carrying out liquid fermentation culture on bacillus cereus, rhodobacter sulfidophilus and streptomyces microflavus to obtain fermentation liquor, adding a mixture of 2kg of modified gelatin and cyclodextrin which are mixed by equal weight into 10kg of bacillus cereus fermentation liquor, emulsifying, and carrying out spray drying to obtain bacillus cereus powder; adding 2kg of a mixture of modified gelatin and cyclodextrin which are mixed by equal weight into 10kg of rhodobacter sulfidophilus fermentation liquor, emulsifying, and performing spray drying to obtain rhodobacter sulfidophilus powder; adding 2kg of cyclodextrin into 10kg of streptomyces microflavus fermentation liquor for emulsification, and performing spray drying to obtain rhodobacter sulfidophilus powder;
(2) preparation of Carrier Dispersion
Mixing 17kg of crop straw powder and 8.5kg of modified medical stone powder uniformly, and adding water to 50kg of the mixture to prepare mixed carrier dispersion liquid;
(3) preparation of composite microbial preparation
Mixing 7kg of waxy bacillus powder, 3kg of sulphur-philic rhodobacter parvum powder and 2kg of streptomyces microflavus powder in the step (1) to obtain mixed bacteria powder; and (3) adding the mixed bacterial powder into the mixed carrier dispersion liquid in the step (2), uniformly stirring, and drying to obtain the composite microbial preparation.
Comparative example 1
Comparative example 1 was prepared in substantially the same manner as in example 5 except that the mixed powder of Streptomyces microflavus in step (3) was replaced with an equal amount of Haematococcus sulfidophilus powder.
Comparative example 2
Comparative example 2 was prepared in substantially the same manner as in example 5, except that the mixed powder of Streptomyces microflavus in step (3) was replaced with an equal amount of Bacillus cereus powder.
Comparative example 3
Comparative example 3 the same procedure as in example 5 was followed except that the Rhodococcus sulfidophilus powder in the mixed powder in step (3) was replaced with an equal amount of Bacillus cereus powder.
Comparative example 4
Comparative example 4 the same procedure as in example 5 was conducted except that the S.sulfidophilus powder in the mixed powder in step (3) was replaced with an equal amount of Streptomyces microflavus powder.
Comparative example 5
Comparative example 5 was prepared in substantially the same manner as in example 5 except that the waxy Bacillus powder in the mixed powder in step (3) was replaced with an equal amount of Streptomyces microflavus powder.
Comparative example 6
Comparative example 6 was prepared in substantially the same manner as in example 5 except that the Bacillus cereus powder in the mixed powder in step (3) was replaced with an equal amount of Haematococcus thiophosphaeri powder.
Comparative example 7
Comparative example 7 was prepared in substantially the same manner as in example 5, except that only one waxy Bacillus powder was contained in the mixed powder in step (3).
Comparative example 8
Comparative example 8 (basically the same preparation method as in example 5 except that only one kind of Rhodotorula sulfidophila powder was contained in the mixed powder in step (3)).
Comparative example 9
Comparative example 9 (basically the same preparation method as example 5 except that only one kind of Streptomyces microflavus powder was contained in the mixed powder in step (3)).
Comparative example 10
Comparative example 10 the same procedure as in example 5 was conducted except that the modified gelatin in step (1) was replaced with a conventional gelatin; the modified medical stone in the step (2) is replaced by conventional medical stone.
Evaluation of prevention and treatment effect of compound microbial preparation on root-knot nematode
The test crop is cucumber, and the test product is the compound microbial preparation prepared in the examples 2-8 and the comparative examples 1-10 of the invention;
the test field is farmland soil in which the cucumbers planted for a long time are damaged by the root-knot nematodes;
the test method comprises the following specific steps: 5 days before the cultivation of the cucumbers, when soil preparation treatment is carried out, the compound microbial preparation is applied to a field by flushing, and 10L (diluted by 400 times) of microbial preparation dispersion liquid is applied to each mu; applying 20L of microbial preparation dispersion liquid (diluted by 600 times) per mu at the seedling stage of the cucumbers; applying 20L of microbial preparation dispersion liquid (diluted by 800 times) to each mu of cucumber in the initial flowering period; applying 20L of microbial preparation dispersion (diluted by 800 times) in the cucumber fruiting period; until the cucumber is pulled out, no microbial preparation is applied; the test groups are subjected to the same field management except that the microbial preparations are applied differently until the cucumbers pull out, and when the test groups are subjected to the compound microbial preparation, the control groups are applied with the same amount of water; and (3) detecting the root-knot nematodes in the soil of the test group and the control group after the cucumber seedlings are pulled out, and evaluating the control effect of the microbial preparation on the root-knot nematodes. The cucumber yield of each group was analyzed, and the results are shown in table 1.
Table 1.
| Root knot nematode Density (strips/g) | Cucumber yield (%) | |
| Control group | 13.52 | / |
| Example 2 | 3.43 | 18.7％ |
| Example 3 | 3.86 | 18.2％ |
| Example 4 | 3.03 | 19.8％ |
| Example 5 | 2.28 | 22.3％ |
| Example 6 | 2.73 | 21.4％ |
| Example 7 | 2.96 | 20.8％ |
| Example 8 | 3.27 | 19.2％ |
| Comparative example 1 | 5.23 | 12.1％ |
| Comparative example 2 | 4.34 | 15.1％ |
| Comparative example 3 | 5.46 | 11.8％ |
| Comparative example 4 | 4.73 | 14.3％ |
| Comparative example 5 | 7.21 | 8.5％ |
| Comparative example 6 | 5.11 | 12.8％ |
| Comparative example 7 | 6.52 | 10.7％ |
| Comparative example 8 | 8.13 | 7.4％ |
| Comparative example 9 | 8.87 | 5.6％ |
| Comparative example 10 | 6.97 | 10.2％ |
As can be seen from the data in table 1, the microbial composite preparation prepared in embodiments 2 to 8 of the present invention has a good control effect on the root-knot nematodes in the soil, and compared with the control group, the density of the root-knot nematodes in the soil is significantly reduced, and the reduction rate reaches more than 71%; the composite preparation formed by mixing the waxy bacillus powder, the sulphur-tolerant rhodobacter xylinum powder and the streptomyces microflavus powder shows a synergistic effect in the aspect of preventing and treating the root-knot nematodes, and when the weight ratio of the waxy bacillus powder, the sulphur-tolerant rhodobacter xylinum powder and the streptomyces microflavus powder is 6:4:2, the prevention and treatment effect on the root-knot nematodes is optimal, and the yield of cucumbers can be obviously improved.
Compared with example 5, the control effect of the compound microbial preparation prepared in comparative examples 1 to 10 on root-knot nematodes and the yield increase rate of cucumbers are reduced, so that the control effect is obviously affected by the proportion of the three bacterial powders, and the carrier selection of the modified medical stone powder has a promoting effect on the control of the root-knot nematodes by the compound microbial preparation.
The above experimental examples are only preferred embodiments of the present invention, and the scope of the present invention is not limited thereto, and any simple changes or equivalent substitutions of the technical solutions that can be obviously obtained by those skilled in the art within the technical scope of the present invention are within the scope of the present invention.
Claims (10)
1. A compound microbial preparation for preventing and treating root-knot nematode is characterized by comprising waxy bacillus powder, sulphur-philic rhodobacter xylinum powder, streptomyces microflavus powder and a carrier; the carrier consists of crop straw powder and modified medical stone powder; the preparation method specifically comprises the following steps:
(1) preparing microbial powder:
respectively carrying out liquid fermentation culture on bacillus cereus, rhodobacter sulfidophilus and streptomyces microflavus to obtain fermentation liquor, adding a protective agent A into the bacillus cereus fermentation liquor, carrying out emulsification treatment, and carrying out spray drying to obtain bacillus cereus powder; adding a protective agent A into the rhodobacter sulfidophilus fermentation liquor, emulsifying, and then carrying out spray drying to obtain rhodobacter sulfidophilus powder; adding a protective agent B into the streptomyces microflavus fermentation liquor, emulsifying, and then carrying out spray drying to obtain rhodobacter sulfidophilus powder;
(2) preparation of Carrier Dispersion
Uniformly mixing crop straw powder and modified medical stone powder, and then adding water to prepare a mixed carrier dispersion liquid with the mass percentage of 50-55%;
(3) preparation of composite microbial preparation
Mixing the waxy bacillus powder, the sulphur-philic rhodobacter parvum powder and the streptomyces microflavus powder in the step (1) to obtain mixed bacteria powder; and (3) adding the mixed bacterial powder into the mixed carrier dispersion liquid in the step (2), uniformly stirring, and drying to obtain the compound microbial agent.
2. The complex microbial preparation for controlling root-knot nematodes according to claim 1, wherein the protective agent A in the step (1) consists of modified gelatin and cyclodextrin in a weight ratio of 1: 1; the protective agent B is cyclodextrin; the addition amount of the protective agent A accounts for 20 percent of the weight of the bacillus cereus fermentation liquor and the rhodobacter sulfidophilus fermentation liquor; the addition amount of the protective agent B accounts for 20 percent of the weight of the streptomyces microflavus fermentation liquor.
3. The complex microbial preparation for controlling Meloidogyne according to claim 1, wherein the weight ratio of the waxy Bacillus powder, the Rhodotorula sulfidophila powder and the Streptomyces microflavus powder in step (2) is 5-7: 3-5: 1-3.
4. The complex microbial preparation for controlling meloidogyne according to claim 1, wherein the weight ratio of the waxy bacillus powder, the rhodobacter sulfidophilus powder and the streptomyces microflavus powder in the step (2) is 6:4: 2.
5. The compound microbial preparation for controlling root-knot nematodes according to claim 1, wherein the weight ratio of the crop straw powder to the modified medical stone powder in the step (2) is 2: 1; the weight ratio of the adding amount of the mixed bacterial powder in the step (3) to the mixed carrier dispersion liquid is 6: 25.
6. The compound microbial preparation for controlling root-knot nematodes as claimed in claim 1, wherein the modified medical stone is a mercapto-modified medical stone, and the preparation method comprises the following steps:
1) sequentially carrying out acid leaching on medical stone, washing with water until washing liquid is neutral, drying, and grinding to obtain medical stone powder;
2) dispersing medical stone in an ethanol solution, and dropwise adding a sulfhydryl modifier into the ethanol solution; heating and stirring at 50-60 deg.C for 2-3 hr, filtering, washing, drying, and grinding to obtain sulfhydryl modified Maifanitum; wherein the mercapto modifier is mercaptopropyltrimethoxysilane.
7. The complex microbial preparation for controlling root-knot nematodes of claim 6, wherein the concentration of said ethanol solution is 50%; the weight volume ratio of the medical stone to the ethanol solution is 50 g/L; the weight ratio of the added amount of the sulfhydryl modifier to the medical stone is 1: 5.
8. The complex microbial preparation for controlling root-knot nematodes according to claim 1, wherein the modified gelatin is gelatin treated with gelatinase, and specifically, the modified gelatin is obtained by adding water to gelatin, stirring for swelling, stirring at 90 ℃ until the gelatin is completely dissolved, cooling to room temperature, adding gelatinase thereto, stirring for reaction at room temperature, and performing enzyme inactivation treatment.
9. The complex microbial preparation for controlling meloidogyne according to claim 1, wherein the number of active viable bacteria in the complex microbial preparation is not less than 0.2 hundred million/g.
10. The use of the compound microbial inoculant for controlling root-knot nematodes according to any one of claims 1 to 9, wherein any one of drip irrigation, flushing application, broadcasting application and root irrigation can be selected as the use method, and the use amount is 10-20 kg/mu.
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