CN117106650A - Agricultural compound microbial agent - Google Patents
Agricultural compound microbial agent Download PDFInfo
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- CN117106650A CN117106650A CN202311091484.0A CN202311091484A CN117106650A CN 117106650 A CN117106650 A CN 117106650A CN 202311091484 A CN202311091484 A CN 202311091484A CN 117106650 A CN117106650 A CN 117106650A
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- 150000001875 compounds Chemical class 0.000 title claims abstract description 34
- 230000000813 microbial effect Effects 0.000 title claims description 8
- 238000000855 fermentation Methods 0.000 claims abstract description 121
- 230000004151 fermentation Effects 0.000 claims abstract description 121
- 235000013399 edible fruits Nutrition 0.000 claims abstract description 53
- 244000063299 Bacillus subtilis Species 0.000 claims abstract description 46
- 235000014469 Bacillus subtilis Nutrition 0.000 claims abstract description 46
- 241000193830 Bacillus <bacterium> Species 0.000 claims abstract description 44
- 241000881860 Paenibacillus mucilaginosus Species 0.000 claims abstract description 44
- 230000000694 effects Effects 0.000 claims abstract description 37
- 239000002068 microbial inoculum Substances 0.000 claims abstract description 26
- 240000008067 Cucumis sativus Species 0.000 claims abstract description 17
- 235000002566 Capsicum Nutrition 0.000 claims abstract description 12
- 239000007788 liquid Substances 0.000 claims abstract description 7
- 240000008574 Capsicum frutescens Species 0.000 claims abstract description 5
- CSNNHWWHGAXBCP-UHFFFAOYSA-L Magnesium sulfate Chemical compound [Mg+2].[O-][S+2]([O-])([O-])[O-] CSNNHWWHGAXBCP-UHFFFAOYSA-L 0.000 claims description 18
- 239000002054 inoculum Substances 0.000 claims description 15
- 240000006365 Vitis vinifera Species 0.000 claims description 14
- 230000012010 growth Effects 0.000 claims description 13
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 13
- VTYYLEPIZMXCLO-UHFFFAOYSA-L Calcium carbonate Chemical compound [Ca+2].[O-]C([O-])=O VTYYLEPIZMXCLO-UHFFFAOYSA-L 0.000 claims description 12
- 239000001963 growth medium Substances 0.000 claims description 11
- 235000010799 Cucumis sativus var sativus Nutrition 0.000 claims description 10
- 239000003795 chemical substances by application Substances 0.000 claims description 10
- ZPWVASYFFYYZEW-UHFFFAOYSA-L dipotassium hydrogen phosphate Chemical compound [K+].[K+].OP([O-])([O-])=O ZPWVASYFFYYZEW-UHFFFAOYSA-L 0.000 claims description 9
- 229910052943 magnesium sulfate Inorganic materials 0.000 claims description 9
- 235000019341 magnesium sulphate Nutrition 0.000 claims description 9
- 229920002261 Corn starch Polymers 0.000 claims description 6
- 239000001888 Peptone Substances 0.000 claims description 6
- 108010080698 Peptones Proteins 0.000 claims description 6
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 claims description 6
- 235000019764 Soybean Meal Nutrition 0.000 claims description 6
- CZMRCDWAGMRECN-UGDNZRGBSA-N Sucrose Chemical compound O[C@H]1[C@H](O)[C@@H](CO)O[C@@]1(CO)O[C@@H]1[C@H](O)[C@@H](O)[C@H](O)[C@@H](CO)O1 CZMRCDWAGMRECN-UGDNZRGBSA-N 0.000 claims description 6
- 229930006000 Sucrose Natural products 0.000 claims description 6
- 235000009754 Vitis X bourquina Nutrition 0.000 claims description 6
- 235000012333 Vitis X labruscana Nutrition 0.000 claims description 6
- 235000014787 Vitis vinifera Nutrition 0.000 claims description 6
- BFNBIHQBYMNNAN-UHFFFAOYSA-N ammonium sulfate Chemical compound N.N.OS(O)(=O)=O BFNBIHQBYMNNAN-UHFFFAOYSA-N 0.000 claims description 6
- 229910052921 ammonium sulfate Inorganic materials 0.000 claims description 6
- 235000011130 ammonium sulphate Nutrition 0.000 claims description 6
- 229910000019 calcium carbonate Inorganic materials 0.000 claims description 6
- 239000008120 corn starch Substances 0.000 claims description 6
- 235000019319 peptone Nutrition 0.000 claims description 6
- 239000004455 soybean meal Substances 0.000 claims description 6
- 239000005720 sucrose Substances 0.000 claims description 6
- 208000031888 Mycoses Diseases 0.000 claims description 4
- 239000001390 capsicum minimum Substances 0.000 claims description 4
- 240000007594 Oryza sativa Species 0.000 claims description 3
- 235000007164 Oryza sativa Nutrition 0.000 claims description 3
- 240000004808 Saccharomyces cerevisiae Species 0.000 claims description 3
- 240000008042 Zea mays Species 0.000 claims description 3
- 235000005824 Zea mays ssp. parviglumis Nutrition 0.000 claims description 3
- 235000002017 Zea mays subsp mays Nutrition 0.000 claims description 3
- 229940041514 candida albicans extract Drugs 0.000 claims description 3
- 235000005822 corn Nutrition 0.000 claims description 3
- 239000013530 defoamer Substances 0.000 claims description 3
- 235000013312 flour Nutrition 0.000 claims description 3
- 229910000402 monopotassium phosphate Inorganic materials 0.000 claims description 3
- 235000019796 monopotassium phosphate Nutrition 0.000 claims description 3
- PJNZPQUBCPKICU-UHFFFAOYSA-N phosphoric acid;potassium Chemical compound [K].OP(O)(O)=O PJNZPQUBCPKICU-UHFFFAOYSA-N 0.000 claims description 3
- 239000000843 powder Substances 0.000 claims description 3
- 235000009566 rice Nutrition 0.000 claims description 3
- 239000011780 sodium chloride Substances 0.000 claims description 3
- 239000012138 yeast extract Substances 0.000 claims description 3
- 230000001737 promoting effect Effects 0.000 claims description 2
- 239000000306 component Substances 0.000 claims 4
- 239000002552 dosage form Substances 0.000 claims 1
- 230000000855 fungicidal effect Effects 0.000 claims 1
- 239000000417 fungicide Substances 0.000 claims 1
- 230000002401 inhibitory effect Effects 0.000 claims 1
- 239000012533 medium component Substances 0.000 claims 1
- 201000010099 disease Diseases 0.000 abstract description 51
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 abstract description 51
- 241000219094 Vitaceae Species 0.000 abstract description 4
- 235000021021 grapes Nutrition 0.000 abstract description 4
- 241000758706 Piperaceae Species 0.000 abstract description 3
- 239000000126 substance Substances 0.000 abstract description 2
- 235000009849 Cucumis sativus Nutrition 0.000 abstract 1
- 238000002360 preparation method Methods 0.000 abstract 1
- 235000010633 broth Nutrition 0.000 description 41
- 241000196324 Embryophyta Species 0.000 description 33
- 239000007633 bacillus mucilaginosus Substances 0.000 description 23
- 230000000052 comparative effect Effects 0.000 description 14
- 241000722363 Piper Species 0.000 description 10
- 235000013339 cereals Nutrition 0.000 description 9
- 239000000203 mixture Substances 0.000 description 9
- 238000002474 experimental method Methods 0.000 description 6
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- 235000017804 Piper guineense Nutrition 0.000 description 5
- 235000008184 Piper nigrum Nutrition 0.000 description 5
- 241000233647 Phytophthora nicotianae var. parasitica Species 0.000 description 4
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- 239000002689 soil Substances 0.000 description 4
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- 230000017260 vegetative to reproductive phase transition of meristem Effects 0.000 description 4
- 241000221785 Erysiphales Species 0.000 description 3
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- 235000002637 Nicotiana tabacum Nutrition 0.000 description 3
- 230000004913 activation Effects 0.000 description 3
- 230000018109 developmental process Effects 0.000 description 3
- 230000006806 disease prevention Effects 0.000 description 3
- 239000003337 fertilizer Substances 0.000 description 3
- 235000012055 fruits and vegetables Nutrition 0.000 description 3
- 239000002609 medium Substances 0.000 description 3
- 208000024891 symptom Diseases 0.000 description 3
- 238000004383 yellowing Methods 0.000 description 3
- 240000000560 Citrus x paradisi Species 0.000 description 2
- 241000233679 Peronosporaceae Species 0.000 description 2
- 230000003213 activating effect Effects 0.000 description 2
- 230000003321 amplification Effects 0.000 description 2
- 230000000443 biocontrol Effects 0.000 description 2
- 239000013043 chemical agent Substances 0.000 description 2
- 238000000034 method Methods 0.000 description 2
- 238000003199 nucleic acid amplification method Methods 0.000 description 2
- 241000894007 species Species 0.000 description 2
- 230000001629 suppression Effects 0.000 description 2
- 235000013311 vegetables Nutrition 0.000 description 2
- 108091000130 1-aminocyclopropane-1-carboxylate deaminase Proteins 0.000 description 1
- 241000193744 Bacillus amyloliquefaciens Species 0.000 description 1
- 102000012286 Chitinases Human genes 0.000 description 1
- 108010022172 Chitinases Proteins 0.000 description 1
- 108090000790 Enzymes Proteins 0.000 description 1
- 102000004190 Enzymes Human genes 0.000 description 1
- -1 IAA Chemical class 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
- 241000109294 Rosa suffulta Species 0.000 description 1
- 241000221662 Sclerotinia Species 0.000 description 1
- XUIMIQQOPSSXEZ-UHFFFAOYSA-N Silicon Chemical compound [Si] XUIMIQQOPSSXEZ-UHFFFAOYSA-N 0.000 description 1
- 241001052560 Thallis Species 0.000 description 1
- 238000012271 agricultural production Methods 0.000 description 1
- 239000003242 anti bacterial agent Substances 0.000 description 1
- 229940088710 antibiotic agent Drugs 0.000 description 1
- 229910052586 apatite Inorganic materials 0.000 description 1
- 244000052616 bacterial pathogen Species 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 230000000975 bioactive effect Effects 0.000 description 1
- 238000012258 culturing Methods 0.000 description 1
- 239000003814 drug Substances 0.000 description 1
- 230000004720 fertilization Effects 0.000 description 1
- 235000011389 fruit/vegetable juice Nutrition 0.000 description 1
- 239000005556 hormone Substances 0.000 description 1
- 229940088597 hormone Drugs 0.000 description 1
- 230000005764 inhibitory process Effects 0.000 description 1
- 229910052500 inorganic mineral Inorganic materials 0.000 description 1
- 230000009545 invasion Effects 0.000 description 1
- 238000011031 large-scale manufacturing process Methods 0.000 description 1
- 238000004519 manufacturing process Methods 0.000 description 1
- 239000010445 mica Substances 0.000 description 1
- 229910052618 mica group Inorganic materials 0.000 description 1
- 244000005700 microbiome Species 0.000 description 1
- 239000011707 mineral Substances 0.000 description 1
- 235000010755 mineral Nutrition 0.000 description 1
- 231100000252 nontoxic Toxicity 0.000 description 1
- 230000003000 nontoxic effect Effects 0.000 description 1
- VSIIXMUUUJUKCM-UHFFFAOYSA-D pentacalcium;fluoride;triphosphate Chemical compound [F-].[Ca+2].[Ca+2].[Ca+2].[Ca+2].[Ca+2].[O-]P([O-])([O-])=O.[O-]P([O-])([O-])=O.[O-]P([O-])([O-])=O VSIIXMUUUJUKCM-UHFFFAOYSA-D 0.000 description 1
- 239000000447 pesticide residue Substances 0.000 description 1
- 239000011574 phosphorus Substances 0.000 description 1
- 229910052698 phosphorus Inorganic materials 0.000 description 1
- 230000029553 photosynthesis Effects 0.000 description 1
- 238000010672 photosynthesis Methods 0.000 description 1
- 230000008635 plant growth Effects 0.000 description 1
- 239000003375 plant hormone Substances 0.000 description 1
- 239000011591 potassium Substances 0.000 description 1
- 229910052700 potassium Inorganic materials 0.000 description 1
- 239000010703 silicon Substances 0.000 description 1
- 229910052710 silicon Inorganic materials 0.000 description 1
- 239000000243 solution Substances 0.000 description 1
- 238000006467 substitution reaction Methods 0.000 description 1
Classifications
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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
- 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
- A01G—HORTICULTURE; CULTIVATION OF VEGETABLES, FLOWERS, RICE, FRUIT, VINES, HOPS OR SEAWEED; FORESTRY; WATERING
- A01G7/00—Botany in general
- A01G7/06—Treatment of growing trees or plants, e.g. for preventing decay of wood, for tingeing flowers or wood, for prolonging the life of 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
- A01N63/22—Bacillus
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01P—BIOCIDAL, PEST REPELLANT, PEST ATTRACTANT OR PLANT GROWTH REGULATORY ACTIVITY OF CHEMICAL COMPOUNDS OR PREPARATIONS
- A01P21/00—Plant growth regulators
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- A—HUMAN NECESSITIES
- 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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- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12R—INDEXING SCHEME ASSOCIATED WITH SUBCLASSES C12C - C12Q, RELATING TO MICROORGANISMS
- C12R2001/00—Microorganisms ; Processes using microorganisms
- C12R2001/01—Bacteria or Actinomycetales ; using bacteria or Actinomycetales
- C12R2001/07—Bacillus
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12R—INDEXING SCHEME ASSOCIATED WITH SUBCLASSES C12C - C12Q, RELATING TO MICROORGANISMS
- C12R2001/00—Microorganisms ; Processes using microorganisms
- C12R2001/01—Bacteria or Actinomycetales ; using bacteria or Actinomycetales
- C12R2001/07—Bacillus
- C12R2001/125—Bacillus subtilis ; Hay bacillus; Grass bacillus
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- Life Sciences & Earth Sciences (AREA)
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- Wood Science & Technology (AREA)
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- Zoology (AREA)
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- Environmental Sciences (AREA)
- General Health & Medical Sciences (AREA)
- Biotechnology (AREA)
- Plant Pathology (AREA)
- Pest Control & Pesticides (AREA)
- Microbiology (AREA)
- Bioinformatics & Cheminformatics (AREA)
- General Chemical & Material Sciences (AREA)
- Forests & Forestry (AREA)
- Ecology (AREA)
- Biodiversity & Conservation Biology (AREA)
- Organic Chemistry (AREA)
- Botany (AREA)
- Virology (AREA)
- Genetics & Genomics (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Medicinal Chemistry (AREA)
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- Tropical Medicine & Parasitology (AREA)
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Abstract
The invention belongs to the technical field of biological control of crops, and discloses an agricultural compound microbial inoculum. The agricultural compound microbial inoculum comprises the following effective components: the bacillus bailii fermentation liquid, the paenibacillus mucilaginosus fermentation liquid and the bacillus subtilis fermentation liquid are mixed according to the volume ratio of 1-5:1:1-5. The agricultural compound microbial inoculum is used for cucumbers, peppers and grapes, and the disease control effects respectively reach 91.34%, 93.02% and 90.58%. The average single-fruit weight of cucumber plants is increased by 41.25g compared with that of a control group, and the average single-plant fruiting amount is increased by 7.06 compared with that of the control group; the average single fruit weight of the capsicum plants is increased by 8.25g compared with that of the control group, and the average single fruit bearing amount is increased by 20.66 compared with that of the control group; the average ear weight and average grain weight for the grapes were increased by 213.25g and 3.66g, respectively, compared to the control group. On the other hand, the agricultural compound microbial inoculum provided by the invention has low preparation cost, is safer than the traditional chemical control, has no pollution to the environment, and can be industrially produced.
Description
Technical Field
The invention belongs to the technical field of biological control of crops, relates to an agricultural compound microbial inoculum, and in particular relates to an agricultural compound microbial inoculum for improving fruit quality and reducing crop diseases.
Background
The production of fruits and vegetables is large in China, and the yield of fruits and vegetables is greatly increased along with the development of market economy. Improving fruit quality such as single fruit weight is a key to improving market competitiveness of fruits and vegetables. At present, for improving the quality of fruits, water and fertilizer management, fruit thinning, bagging, fertilization, leaf picking, fruit turning and the like are mainly adopted.
Bacillus bailii (bacillus velezensis) is a widely used biological control strain and is excellent in control of various fungal diseases. Patent publication No. CN115011503A discloses Bacillus belicus for controlling tobacco black shank and phytophthora nicotianae. In addition, bacillus beleiensis promotes fruit growth by producing hormones and volatile compounds, such as IAA, NH, and ACC deaminase, through the vital activities of the thalli.
Bacillus mucilaginosus (Bacillus mucilaginosus) is an aerobic or facultative anaerobic bacillus-producing bacillus which can decompose minerals such as mica and apatite and release elements such as potassium, silicon and phosphorus when applied to soil as fertilizer, thereby increasing soil fertility. In addition, the paenibacillus mucilaginosus can also produce various bioactive substances such as various antibiotics, enzymes, plant hormones and the like, has good inhibition effect on pathogenic bacteria of plants, and simultaneously promotes the growth of crops.
Bacillus subtilis (Bacillus subtilis) is widely distributed in soil and spoilage organisms and is susceptible to propagation in the juice of the herb. Bacillus subtilis can secrete a large amount of chitinase for purifying water quality. In addition, the strain is also used for preventing and controlling plant diseases, for example, the patent with publication number CN102747103A discloses a biocontrol strain capable of resisting both tobacco black shank and bacterial wilt, and the prevention and control effect of field application on tobacco black shank and bacterial wilt mixed fuming field is 61.04%.
On the other hand, chemical agents are still the mainstream plant disease control methods at present, and it is still difficult to realize efficient and safe control of plant diseases by means of chemical agents or single-strain biocontrol microbial agents. Therefore, a compound microbial inoculum capable of preventing and treating plant diseases and promoting plant growth or improving fruit quality is needed to meet the demands of agricultural production.
Disclosure of Invention
Based on the technical problems, the invention provides an agricultural compound microbial inoculum, and the bacterial species of the agricultural compound microbial inoculum are bacillus belicus, paenibacillus mucilaginosus and bacillus subtilis.
Further, the agricultural compound microbial inoculum comprises the following effective components: a combination of bacillus belicus fermentation broth, paenibacillus mucilaginosus fermentation broth and bacillus subtilis fermentation broth; the effective viable count of the effective component is more than or equal to 5 multiplied by 10 8 And each mL.
Further, the bacillus subtilis fermentation liquor, the paenibacillus mucilaginosus fermentation liquor and the bacillus subtilis fermentation liquor are combined according to the following proportion: the volume ratio is 1-5:1:1-5.
Further, the culture medium for preparing the bacillus beijerinus fermentation broth comprises the following components: the corn starch comprises, by mass, 4 parts of corn starch, 4 parts of soybean meal, 0.2 part of yeast powder, 1.5 parts of sucrose, 0.2 part of peptone, 0.2 part of dipotassium hydrogen phosphate, 0.4 part of potassium dihydrogen phosphate, 0.2 part of sodium chloride, 0.1 part of calcium carbonate, 0.4 part of defoamer, pH of which is adjusted to 7.0, and the balance of water;
the fermentation parameters are as follows: the temperature is 37 ℃, the rotating speed is 180r/min, the inoculum size of bacillus subtilis is 5 percent, and the fermentation time is 72 hours;
further, the culture medium for preparing the paenibacillus mucilaginosus fermentation broth comprises the following components: 3 parts of sucrose, 3 parts of peptone, 3 parts of dipotassium hydrogen phosphate, 2.8 parts of yeast extract, 1.4 parts of ammonium sulfate, 1.4 parts of magnesium sulfate, and the balance of water, wherein the pH value of the magnesium sulfate is adjusted to 7.2-7.4;
the fermentation parameters are as follows: the temperature is 32 ℃, the rotating speed is 180r/min, the inoculum size of the paenibacillus mucilaginosus is 5 percent, and the fermentation time is 72 hours;
further, the culture medium for preparing the bacillus subtilis fermentation broth comprises the following components: 55 parts of rice hulls, 25 parts of corn flour, 15 parts of soybean meal, 5 parts of bran, 0.06 part of dipotassium hydrogen phosphate, 0.015 part of magnesium sulfate, 0.03 part of ammonium sulfate, 0.195 part of light calcium carbonate and the balance of water;
the fermentation parameters are as follows: the temperature is 30 ℃, the relative humidity is 85%, the bacillus subtilis inoculum size is 10%, and the fermentation time is 72 hours.
On the other hand, the invention claims the application of the agricultural compound microbial inoculum in improving the fruit quality or reducing the crop fungal diseases.
As a preferred embodiment of the present invention, the crop comprises: cucumber, capsicum, grape.
Compared with the prior art, the technical scheme provided by the invention has the following beneficial effects:
the agricultural compound microbial inoculum provided by the invention is obtained by applying a test to cucumber plants, the disease control effect reaches 91.34%, the average single fruit weight is 255.13g, 41.25g is improved compared with a control group, the average single fruit bearing amount is 20.97, and 7.06 is improved compared with the control group.
The agricultural compound microbial inoculum provided by the invention is obtained by applying experiments to pepper plants, the disease control effect reaches 93.02%, the average single fruit weight is 23.89g, 8.25g is improved compared with a control group, the average single plant fruiting amount is 45.17, and 20.66 is improved compared with the control group.
The agricultural compound microbial inoculum provided by the invention is obtained by applying experiments to grape plants, the disease control effect reaches 90.58%, the average spike weight is 733.58g, 213.25g is increased compared with a control group, the average grain weight is 12.62g, and 3.66g is increased compared with the control group.
The agricultural compound microbial inoculum provided by the invention is nontoxic and harmless to the environment, does not generate pesticide residues, pollutes water or soil, has low fermentation cost, is simple to prepare and easy to operate, can meet the requirement of large-scale production, and is suitable for popularization and use.
Detailed Description
The present invention will be described in further detail with reference to the following examples in order to make the objects, technical solutions and advantages of the present invention more apparent. It should be understood that the detailed description and specific examples are intended for purposes of illustration only and are not intended to limit the scope of the invention.
In various embodiments, the medium used for bacillus beijerinus activation is medium No. one of gao; the culture medium used for activating the paenibacillus mucilaginosus is a PDA flat plate; the culture medium used for activating the bacillus subtilis is a PDA plate.
In various embodiments, the culture medium composition for preparing the bacillus beijerinus fermentation broth is: the corn starch comprises, by mass, 4 parts of corn starch, 4 parts of soybean meal, 0.2 part of yeast powder, 1.5 parts of sucrose, 0.2 part of peptone, 0.2 part of dipotassium hydrogen phosphate, 0.4 part of potassium dihydrogen phosphate, 0.2 part of sodium chloride, 0.1 part of calcium carbonate, 0.4 part of defoamer, pH of which is adjusted to 7.0, and the balance of water; the fermentation parameters are as follows: the temperature is 37 ℃, the rotating speed is 180r/min, the inoculum size of the bacillus beijerinckii is 5 percent, and the fermentation time is 72 hours.
The culture medium for preparing the paenibacillus mucilaginosus fermentation broth comprises the following components: 3 parts of sucrose, 3 parts of peptone, 3 parts of dipotassium hydrogen phosphate, 2.8 parts of yeast extract, 1.4 parts of ammonium sulfate, 1.4 parts of magnesium sulfate, and the balance of water, wherein the pH value of the magnesium sulfate is adjusted to 7.2-7.4; the fermentation parameters are as follows: the temperature is 32 ℃, the rotating speed is 180r/min, the inoculum size of the paenibacillus mucilaginosus is 5 percent, and the fermentation time is 72 hours.
The culture medium for preparing the bacillus subtilis fermentation broth comprises the following components: 55 parts of rice hulls, 25 parts of corn flour, 15 parts of soybean meal, 5 parts of bran, 0.06 part of dipotassium hydrogen phosphate, 0.015 part of magnesium sulfate, 0.03 part of ammonium sulfate, 0.195 part of light calcium carbonate and the balance of water; the fermentation parameters are as follows: the temperature is 30 ℃, the relative humidity is 85%, the bacillus subtilis inoculum size is 10%, and the fermentation time is 72 hours.
The bacillus belicus, paenibacillus mucilaginosus and bacillus subtilis strains are purchased from Shanghai morning microorganisms.
Example 1
This example provides for the activation and amplification culture of species.
The deposited Bacillus belicus, paenibacillus mucilaginosus and Bacillus subtilis were inoculated onto the activation medium, respectively, using a plate streaking method. Culturing at 28 deg.c for 2-3 d to obtain the activated strain.
The activated strain is subjected to the culture medium groupAnd (5) carrying out amplification culture under different fermentation conditions to obtain bacillus belicus, paenibacillus mucilaginosus and bacillus subtilis fermentation liquor respectively. The cell concentrations were adjusted to 5×10 with an ultraviolet spectrophotometer 8 CFU/mL。
Example 2
This example provides the experimental effect of applying a mixture of 3 fermentation broths to cucumber.
The strain was activated and grown up in the manner described in example 1 before each application, and the mixture was prepared immediately before each application.
The test set up control and test groups, the control group did not administer any agent.
Mixing bacillus bailii fermentation liquor, bacillus mucilaginosus fermentation liquor and bacillus subtilis fermentation liquor according to the volume ratio of 1:1:1 to obtain mixed liquor for test group 1;
mixing bacillus bailii fermentation liquor, bacillus mucilaginosus fermentation liquor and bacillus subtilis fermentation liquor according to the volume ratio of 3:1:3 to obtain mixed liquor for test group 2;
mixing bacillus bailii fermentation liquor, bacillus mucilaginosus fermentation liquor and bacillus subtilis fermentation liquor according to the volume ratio of 1:1:5 to obtain mixed liquor for test group 3;
mixing bacillus bailii fermentation liquor, bacillus mucilaginosus fermentation liquor and bacillus subtilis fermentation liquor according to the volume ratio of 5:1:1 to obtain mixed liquor for test group 4;
the bacillus bailii fermentation broth, the paenibacillus mucilaginosus fermentation broth and the bacillus subtilis fermentation broth are mixed according to the volume ratio of 5:1:5 to obtain a mixed solution for the test group 5.
In the embodiment, the Shuangfeng cucumber is used as a test object, and the test site is the vegetable planting garden in Yu county of Kai city in Henan province. The fertility level of the land block in the park is the same as the management condition, and the tested plants are consistent in growth vigor. The land parcels of the test group and the control group are not adjacent. 200 cucumber seedlings are selected from each test group and control group, and total 1200 cucumber seedlings are selected. Each test group was repeatedly applied 4 times, sprayed for the first time when the plants were grown to a plant height of about 40cm, then applied every 10 days for 40 days, the control group was not applied with any agent, and both the test group and the control group were subjected to normal field water and fertilizer management.
Marking in flowering, taking the fact that the fruit growth speed of the cucumber is high, picking off fruits before and after the 14 th day and 19 th day from the flowering day, recording the number and the weight of the fruits in a test group and a control group, counting average single fruit weight after the test is finished, and counting the morbidity and the prevention effect of each group on the 3 rd day after the last spraying is finished. Wherein,
average single fruit weight = total weight of fruit/total number of fruits;
the disease categories incorporating statistics were: brown spot, downy mildew, leaf blight, anthracnose, target spot, powdery mildew, that is, a plant that is affected if a cucumber plant is infected with any of the above diseases.
The morbidity and the control effect were calculated according to the following formula, and the results are shown in table 1.
Incidence = number of lesions/total number of trial plants x 100%
Control effect= (control disease rate-treatment disease rate)/control disease rate×100%
Table 1 test results of cucumber plants test and control groups
Table 1 shows that the control group had higher disease incidence in the later period without any disease prevention measure, and more than 60% of plants were infected with the disease to different degrees, showing symptoms such as yellowing and withering of leaves of the plants, and gradually involved other plants, and the photosynthesis was affected by the damage of the leaves, resulting in slower fruit growth rate and smaller fruit weight.
The disease expansion of each test group is not obvious, and the overall control effect is obviously improved compared with the control group. In addition, from the average single fruit weight and the average single plant fruit bearing quantity, each test group is obviously improved compared with the control group. Specifically, the disease control effect of test group 5 reached 91.34%. The average single fruit weight is 255.13g, which is improved by 41.25g compared with the control group; the average single plant fruiting amount is 20.97, and 7.06 fruits are added compared with the control group, so that the effect is obvious. Therefore, the compound microbial inoculum provided by the invention can effectively prevent and treat disease attack of cucumber plants, promote cucumber growth and improve fruit quality.
Comparative example 1
This comparative example provides a test effect of applying a mixture of 2 of the fermentation broths to cucumber.
The experimental procedure of this comparative example is identical to that of example 1, except that: the microbial inoculum of the test group was set as follows:
control group: no agent is administered.
Test group 1: bacillus bailii fermentation broth: bacillus mucilaginosus fermentation broth: the volume ratio of the bacillus subtilis fermentation liquor is set as follows: 0:1:1, a step of;
test group 2: bacillus bailii fermentation broth: bacillus mucilaginosus fermentation broth: the volume ratio of the bacillus subtilis fermentation liquor is set as follows: 1:0:1, a step of;
test group 3: bacillus bailii fermentation broth: bacillus mucilaginosus fermentation broth: the volume ratio of the bacillus subtilis fermentation liquor is set as follows: 1:1:0 and the test results are shown in Table 2.
Table 2, comparative example 1 test results
As can be seen from Table 2, reducing one of the components has a certain suppression effect on the disease development of cucumber plants, but the overall control effect is not ideal, and part of plants still have reduced yield due to the disease. Compared with the test result of the comparative example, the compound microbial inoculum provided by the invention has better cucumber disease control effect.
Example 3
This example provides the test effect of applying a mixture of 3 fermentation broths to capsicum.
The strain was activated and grown up in the manner described in example 1 before each application, and the mixture was prepared immediately before each application.
Mixing bacillus bailii fermentation liquor, bacillus mucilaginosus fermentation liquor and bacillus subtilis fermentation liquor according to the volume ratio of 1:1:1 to obtain mixed liquor for test group 1;
mixing bacillus bailii fermentation liquor, bacillus mucilaginosus fermentation liquor and bacillus subtilis fermentation liquor according to the volume ratio of 3:1:3 to obtain mixed liquor for test group 2;
mixing bacillus bailii fermentation liquor, bacillus mucilaginosus fermentation liquor and bacillus subtilis fermentation liquor according to the volume ratio of 1:1:5 to obtain mixed liquor for test group 3;
mixing bacillus bailii fermentation liquor, bacillus mucilaginosus fermentation liquor and bacillus subtilis fermentation liquor according to the volume ratio of 5:1:1 to obtain mixed liquor for test group 4;
the bacillus bailii fermentation broth, the paenibacillus mucilaginosus fermentation broth and the bacillus subtilis fermentation broth are mixed according to the volume ratio of 5:1:5 to obtain a mixed solution for the test group 5.
In the embodiment, peppery Feng third variety of wire peppers are used as test objects, and the test sites and the embodiment 2 are on different plots of the same vegetable planting park. The fertility level of the land block in the park is the same as the management condition, and the tested plants are consistent in growth vigor. The land parcels of the test group and the control group are not adjacent. 200 pepper seedlings are selected from each test group and control group, and total 1200 pepper seedlings are selected. The test group was repeatedly applied 4 times, sprayed for the first time when the plant was budded, then applied every 10 days for 40 days, the control group was not applied with any agent, and normal field management was performed for both the test group and the control group.
Marking when flowering, picking fruits on the 16 th day from the day of flowering, recording the number and weight of the fruits in the test group and the control group, counting average single fruit weight after the test is finished, and counting the morbidity and prevention effect of each group on the 3 rd day after the last spraying is finished.
Wherein,
average single fruit weight = total weight of fruit/total number of fruits;
the disease categories incorporating statistics were: epidemic disease, sclerotinia rot, powdery mildew, brown spot, anthracnose, southern blight, gray mold, that is, a plant infected with any one of the above diseases is designated as an infected plant.
The morbidity and the control effect were calculated according to the following formula, and the results are shown in table 3.
Incidence = number of lesions/total number of trial plants x 100%
Control effect= (control disease rate-treatment disease rate)/control disease rate×100%
TABLE 3 results of experiments with Capsici fructus plants in the test and control groups
Table 3 shows that the control group pepper plants have higher disease incidence rate in the later period without any disease prevention measures according to the test results, and more than 60% of pepper plants are infected with diseases to different degrees, so that symptoms such as complete pepper withering, yellowing or falling of leaves, soft rot of fruits and the like appear. Over time, the disease is also more and more serious, and the infected plants are more and more, so that the fruit quality is seriously affected.
The plant diseases of each test group are effectively controlled, and the overall control effect is good, wherein the disease control effect of the test group 5 reaches 93.02%. In addition, from the average single fruit weight and the average single plant fruit bearing amount, each test group is obviously improved compared with the control group: wherein, the average single fruit weight of the test group 5 reaches 23.89g, which is 8.25g higher than that of the control group; the average single plant fruiting amount is 45.17, which is 20.66 higher than that of the control group. Therefore, the compound microbial inoculum provided by the invention can effectively prevent and treat the disease invasion of pepper plants, promote the growth of peppers and improve the fruit quality.
Comparative example 2
This comparative example provides a test effect of applying a mixture of 2 of the fermentation broths to capsicum.
The experimental procedure of this comparative example is identical to that of example 2, except that: the microbial inoculum of the test group was set as follows:
control group: no agent is administered.
Test group 1: bacillus bailii fermentation broth: bacillus mucilaginosus fermentation broth: the volume ratio of the bacillus subtilis fermentation liquor is set as follows: 0:1:1, a step of;
test group 2: bacillus bailii fermentation broth: bacillus mucilaginosus fermentation broth: the volume ratio of the bacillus subtilis fermentation liquor is set as follows: 1:0:1, a step of;
test group 3: bacillus bailii fermentation broth: bacillus mucilaginosus fermentation broth: the volume ratio of the bacillus subtilis fermentation liquor is set as follows: 1:1:0 and the test results are shown in Table 4.
Table 4, results of comparative example 2
As can be seen from Table 4, reducing one of the components has a certain suppression effect on the disease development of pepper plants, but the overall control effect is not ideal, about 50% -60%, and still some plants are reduced in yield due to the disease. The comparison of the test results of the comparative example and the test results of the example 2 shows that the microbial inoculum provided by the example has better effects of resisting pepper diseases and improving pepper yield, and each component is indispensable.
Example 4
This example provides the effect of applying a mixture of 3 fermentation broths to grapes.
The strain was activated and grown up in the manner described in example 1 before each application, and the mixture was prepared immediately before each application.
Mixing bacillus bailii fermentation liquor, bacillus mucilaginosus fermentation liquor and bacillus subtilis fermentation liquor according to the volume ratio of 1:1:1 to obtain mixed liquor for test group 1;
mixing bacillus bailii fermentation liquor, bacillus mucilaginosus fermentation liquor and bacillus subtilis fermentation liquor according to the volume ratio of 3:1:3 to obtain mixed liquor for test group 2;
mixing bacillus bailii fermentation liquor, bacillus mucilaginosus fermentation liquor and bacillus subtilis fermentation liquor according to the volume ratio of 1:1:5 to obtain mixed liquor for test group 3;
mixing bacillus bailii fermentation liquor, bacillus mucilaginosus fermentation liquor and bacillus subtilis fermentation liquor according to the volume ratio of 5:1:1 to obtain mixed liquor for test group 4;
the bacillus bailii fermentation broth, the paenibacillus mucilaginosus fermentation broth and the bacillus subtilis fermentation broth are mixed according to the volume ratio of 5:1:5 to obtain a mixed solution for the test group 5.
In the embodiment, the sunshine rose variety grape is used as a test object, and the test site is a grape plantation in Yi region of Xishan City of Shanxi province. The fertility level of the land block in the park is the same as the management condition, and the tested plants are consistent in growth vigor. The land parcels of the test group and the control group are not adjacent. 200 grape plants with consistent growth vigor are selected from each test group and control group, and total is 1200. Considering that the grape fruit growth time is longer, the test group is repeatedly applied for 6 times, the first spraying is performed when the plant buds are found, then the test group is applied every 20 days for 120 days, the control group is not applied with any medicament, and the test group and the control group are subjected to normal field management.
And respectively picking fruits of the test group and the control group after the fruits are ripe, recording the total weight of the fruits picked by the grapes of the test group and the control group, the total spike number of the picked fruits and the corresponding grain number of each spike, calculating the average spike weight and the average grain weight after the test is finished, and counting the morbidity and the prevention and treatment effect of each group on the 10 th day after the last spraying is finished. Wherein,
average spike weight = total weight of fruit/total spike number;
average grain weight = total weight of fruit after removal of fruit stalks/total grain number of fruit;
the disease categories incorporating statistics were: downy mildew, anthracnose, white rot, black rot, anthracnose, gray mold, powdery mildew, brown spot, that is, a infected plant if a grape plant is infected with any of the above diseases.
The disease rate and the control effect were calculated according to the following formulas, and the results are shown in table 5.
Incidence = number of lesions/total number of trial plants x 100%
Control effect= (control disease rate-treatment disease rate)/control disease rate×100%
Table 5 test results of grape plant test group and control group
Table 5 shows that the grape plants in the control group have higher disease incidence rate in the later period without any disease prevention measures according to the test results, 69.0% of grape plants are infected with diseases to different degrees, and symptoms such as wilting, rotting, falling or having lesions, yellowing, curling, falling and the like of grape fruits appear in part. Over time, the disease is also more and more serious, and the infected plants are more and more, so that the fruit quality is seriously affected.
The disease expansion of each test group is not obvious, and the overall control effect is better, wherein the disease control effect of the test group 5 reaches 90.58 percent. From the average spike weight and the average grain weight, each test group was significantly improved over the control group: wherein, the average spike weight of the test group 5 reaches 733.58g, which is 213.25g higher than that of the control group; the average grain weight is 12.62g, and the effect is obvious when the grain weight is increased by 3.66g compared with the control group. Therefore, the compound microbial inoculum provided by the invention can effectively prevent and treat disease attack of grape plants, promote grape growth and improve fruit quality.
Comparative example 3
The present comparative example provides a test effect in which a mixed solution of 2 bacterial liquids was applied to grape.
The experimental procedure of this comparative example is identical to that of example 3, except that: the microbial inoculum of the test group was set as follows:
control group: no agent is administered.
Test group 1: bacillus bailii fermentation broth: bacillus mucilaginosus fermentation broth: the volume ratio of the bacillus subtilis fermentation liquor is set as follows: 0:1:1, a step of;
test group 2: bacillus bailii fermentation broth: bacillus mucilaginosus fermentation broth: the volume ratio of the bacillus subtilis fermentation liquor is set as follows: 1:0:1, a step of;
test group 3: bacillus bailii fermentation broth: bacillus mucilaginosus fermentation broth: the volume ratio of the bacillus subtilis fermentation liquor is set as follows: 1:1:0 and the test results are shown in Table 6.
TABLE 6 test results for comparative example 3
As can be seen from Table 6, the effect of the absence of any component on disease control or fruit quality improvement of grape plants was not as good as in example 3, wherein the disease control effect was about 60%, and the average spike weight and average grain weight were also not good. Therefore, the 3 components provided by the invention are indispensable.
The embodiments described above are only some, but not all, embodiments of the invention. The detailed description of the embodiments of the invention is not intended to limit the scope of the invention, as claimed, but is merely representative of selected embodiments of the invention. All other embodiments obtained without inventive effort by a person skilled in the art, which are related deductions and substitutions made by the person skilled in the art under the condition of the inventive concept, are within the scope of protection of the present invention.
Claims (9)
1. The agricultural compound microbial inoculum is characterized in that the adopted strain of the agricultural compound microbial inoculum consists of bacillus belicus, paenibacillus mucilaginosus and bacillus subtilis.
2. The agricultural compound microbial inoculant of claim 1, wherein the agricultural compound microbial inoculant comprises the following effective components: bacillus belicus fermentation liquor, paenibacillus mucilaginosus fermentation liquor and bacillus subtilis fermentation liquor;
the effective viable count of the effective component is more than or equal to 5 multiplied by 10 8 individual/mL;
the agricultural compound microbial inoculum has the effects of promoting fruit growth or inhibiting fungal diseases.
3. The agricultural compound microbial agent according to claim 2, wherein the ratio of the bacillus belicus fermentation liquid, the paenibacillus mucilaginosus fermentation liquid and the bacillus subtilis fermentation liquid is 1-5:1:1-5 in terms of volume ratio.
4. The agricultural compound microbial agent of claim 2, wherein the culture medium components for preparing the bacillus beijerinus fermentation broth are as follows: the corn starch comprises, by mass, 4 parts of corn starch, 4 parts of soybean meal, 0.2 part of yeast powder, 1.5 parts of sucrose, 0.2 part of peptone, 0.2 part of dipotassium hydrogen phosphate, 0.4 part of potassium dihydrogen phosphate, 0.2 part of sodium chloride, 0.1 part of calcium carbonate, 0.4 part of defoamer, pH of which is adjusted to 7.0, and the balance of water;
the fermentation parameters are as follows: the temperature is 37 ℃, the rotating speed is 180r/min, the inoculum size of the bacillus beijerinckii is 5 percent, and the fermentation time is 72 hours.
5. The agricultural compound inoculant of claim 2, wherein the culture medium components for preparing the paenibacillus mucilaginosus fermentation broth are: 3 parts of sucrose, 3 parts of peptone, 3 parts of dipotassium hydrogen phosphate, 2.8 parts of yeast extract, 1.4 parts of ammonium sulfate, 1.4 parts of magnesium sulfate, and the balance of water, wherein the pH value of the magnesium sulfate is adjusted to 7.2-7.4;
the fermentation parameters are as follows: the temperature is 32 ℃, the rotating speed is 180r/min, the inoculum size of the paenibacillus mucilaginosus is 5 percent, and the fermentation time is 72 hours.
6. The agricultural compound microbial inoculant of claim 2, wherein the medium components for preparing the bacillus subtilis broth are: 55 parts of rice hulls, 25 parts of corn flour, 15 parts of soybean meal, 5 parts of bran, 0.06 part of dipotassium hydrogen phosphate, 0.015 part of magnesium sulfate, 0.03 part of ammonium sulfate, 0.195 part of light calcium carbonate and the balance of water;
the fermentation parameters are as follows: the temperature is 30 ℃, the relative humidity is 85%, the bacillus subtilis inoculum size is 10%, and the fermentation time is 72 hours.
7. The use of the agricultural compound fungicide of any one of claims 1 to 6 for improving fruit quality or reducing crop fungal disease.
8. The use according to claim 7, wherein the crop comprises: cucumber, capsicum, grape.
9. The agricultural compound inoculant of any one of claims 1-6, wherein the agricultural compound inoculant is formulated for application after processing into an agriculturally acceptable dosage form.
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