CN117024210A - Microbial composition for preventing and treating fusarium root rot and application thereof - Google Patents
Microbial composition for preventing and treating fusarium root rot and application thereof Download PDFInfo
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- CN117024210A CN117024210A CN202311280139.1A CN202311280139A CN117024210A CN 117024210 A CN117024210 A CN 117024210A CN 202311280139 A CN202311280139 A CN 202311280139A CN 117024210 A CN117024210 A CN 117024210A
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- bacillus
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- 230000000813 microbial effect Effects 0.000 title claims abstract description 58
- 239000000203 mixture Substances 0.000 title claims abstract description 52
- 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 title claims abstract description 48
- 241000223218 Fusarium Species 0.000 title claims abstract description 48
- 239000002689 soil Substances 0.000 claims abstract description 55
- 241000196324 Embryophyta Species 0.000 claims abstract description 50
- CIWBSHSKHKDKBQ-JLAZNSOCSA-N Ascorbic acid Chemical compound OC[C@H](O)[C@H]1OC(=O)C(O)=C1O CIWBSHSKHKDKBQ-JLAZNSOCSA-N 0.000 claims abstract description 40
- 241000193830 Bacillus <bacterium> Species 0.000 claims abstract description 35
- QJZYHAIUNVAGQP-UHFFFAOYSA-N 3-nitrobicyclo[2.2.1]hept-5-ene-2,3-dicarboxylic acid Chemical compound C1C2C=CC1C(C(=O)O)C2(C(O)=O)[N+]([O-])=O QJZYHAIUNVAGQP-UHFFFAOYSA-N 0.000 claims abstract description 26
- 239000004021 humic acid Substances 0.000 claims abstract description 26
- 210000003608 fece Anatomy 0.000 claims abstract description 22
- 235000010323 ascorbic acid Nutrition 0.000 claims abstract description 20
- 229960005070 ascorbic acid Drugs 0.000 claims abstract description 20
- 239000011668 ascorbic acid Substances 0.000 claims abstract description 20
- 241000278457 Bacillus siamensis Species 0.000 claims abstract description 19
- 229910021536 Zeolite Inorganic materials 0.000 claims abstract description 18
- HNPSIPDUKPIQMN-UHFFFAOYSA-N dioxosilane;oxo(oxoalumanyloxy)alumane Chemical compound O=[Si]=O.O=[Al]O[Al]=O HNPSIPDUKPIQMN-UHFFFAOYSA-N 0.000 claims abstract description 18
- 239000000843 powder Substances 0.000 claims abstract description 18
- 239000010457 zeolite Substances 0.000 claims abstract description 18
- 244000269722 Thea sinensis Species 0.000 claims abstract description 16
- 150000007965 phenolic acids Chemical class 0.000 claims abstract description 16
- 240000006497 Dianthus caryophyllus Species 0.000 claims abstract description 12
- 235000009024 Ceanothus sanguineus Nutrition 0.000 claims abstract description 11
- 235000009355 Dianthus caryophyllus Nutrition 0.000 claims abstract description 11
- 240000003553 Leptospermum scoparium Species 0.000 claims abstract description 11
- 235000015459 Lycium barbarum Nutrition 0.000 claims abstract description 11
- 244000288157 Passiflora edulis Species 0.000 claims abstract description 11
- 235000000370 Passiflora edulis Nutrition 0.000 claims abstract description 11
- 238000004321 preservation Methods 0.000 claims abstract description 11
- 230000001276 controlling effect Effects 0.000 claims description 29
- 201000010099 disease Diseases 0.000 claims description 24
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 claims description 24
- 239000003337 fertilizer Substances 0.000 claims description 24
- 235000013616 tea Nutrition 0.000 claims description 15
- 241000207199 Citrus Species 0.000 claims description 12
- VYPSYNLAJGMNEJ-UHFFFAOYSA-N Silicium dioxide Chemical compound O=[Si]=O VYPSYNLAJGMNEJ-UHFFFAOYSA-N 0.000 claims description 12
- 235000020971 citrus fruits Nutrition 0.000 claims description 11
- IJGRMHOSHXDMSA-UHFFFAOYSA-N Atomic nitrogen Chemical compound N#N IJGRMHOSHXDMSA-UHFFFAOYSA-N 0.000 claims description 10
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 9
- 239000000377 silicon dioxide Substances 0.000 claims description 6
- LSHVYAFMTMFKBA-PZJWPPBQSA-N (+)-catechin-3-O-gallate Chemical compound O([C@H]1CC2=C(O)C=C(C=C2O[C@@H]1C=1C=C(O)C(O)=CC=1)O)C(=O)C1=CC(O)=C(O)C(O)=C1 LSHVYAFMTMFKBA-PZJWPPBQSA-N 0.000 claims description 5
- OAICVXFJPJFONN-UHFFFAOYSA-N Phosphorus Chemical compound [P] OAICVXFJPJFONN-UHFFFAOYSA-N 0.000 claims description 5
- ZLMJMSJWJFRBEC-UHFFFAOYSA-N Potassium Chemical compound [K] ZLMJMSJWJFRBEC-UHFFFAOYSA-N 0.000 claims description 5
- XAGFODPZIPBFFR-UHFFFAOYSA-N aluminium Chemical compound [Al] XAGFODPZIPBFFR-UHFFFAOYSA-N 0.000 claims description 5
- 229910052782 aluminium Inorganic materials 0.000 claims description 5
- LVJJFMLUMNSUFN-UHFFFAOYSA-N gallocatechin gallate Natural products C1=C(O)C=C2OC(C=3C=C(O)C(O)=CC=3)C(O)CC2=C1OC(=O)C1=CC(O)=C(O)C(O)=C1 LVJJFMLUMNSUFN-UHFFFAOYSA-N 0.000 claims description 5
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- 239000011574 phosphorus Substances 0.000 claims description 5
- 229910052698 phosphorus Inorganic materials 0.000 claims description 5
- 239000011591 potassium Substances 0.000 claims description 5
- 229910052700 potassium Inorganic materials 0.000 claims description 5
- 229910052500 inorganic mineral Inorganic materials 0.000 claims description 4
- 239000011707 mineral Substances 0.000 claims description 4
- 229910052710 silicon Inorganic materials 0.000 claims description 4
- 239000010703 silicon Substances 0.000 claims description 4
- 244000005700 microbiome Species 0.000 claims description 3
- TWNQGVIAIRXVLR-UHFFFAOYSA-N oxo(oxoalumanyloxy)alumane Chemical compound O=[Al]O[Al]=O TWNQGVIAIRXVLR-UHFFFAOYSA-N 0.000 claims description 3
- 230000001105 regulatory effect Effects 0.000 claims description 3
- 230000000593 degrading effect Effects 0.000 claims description 2
- 235000012239 silicon dioxide Nutrition 0.000 claims description 2
- 239000000126 substance Substances 0.000 abstract description 11
- 230000008485 antagonism Effects 0.000 abstract description 2
- 244000052616 bacterial pathogen Species 0.000 abstract description 2
- WPYMKLBDIGXBTP-UHFFFAOYSA-N benzoic acid Chemical compound OC(=O)C1=CC=CC=C1 WPYMKLBDIGXBTP-UHFFFAOYSA-N 0.000 description 18
- 239000002068 microbial inoculum Substances 0.000 description 14
- 230000000694 effects Effects 0.000 description 13
- 238000002360 preparation method Methods 0.000 description 10
- 239000005711 Benzoic acid Substances 0.000 description 9
- 235000010233 benzoic acid Nutrition 0.000 description 9
- 230000015556 catabolic process Effects 0.000 description 8
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- 239000003795 chemical substances by application Substances 0.000 description 7
- 238000000034 method Methods 0.000 description 7
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- 239000000523 sample Substances 0.000 description 7
- XEKOWRVHYACXOJ-UHFFFAOYSA-N Ethyl acetate Chemical compound CCOC(C)=O XEKOWRVHYACXOJ-UHFFFAOYSA-N 0.000 description 6
- OKKJLVBELUTLKV-UHFFFAOYSA-N Methanol Chemical compound OC OKKJLVBELUTLKV-UHFFFAOYSA-N 0.000 description 6
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- 239000003895 organic fertilizer Substances 0.000 description 5
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- HEMHJVSKTPXQMS-UHFFFAOYSA-M sodium hydroxide Inorganic materials [OH-].[Na+] HEMHJVSKTPXQMS-UHFFFAOYSA-M 0.000 description 5
- 230000009418 agronomic effect Effects 0.000 description 4
- 239000000463 material Substances 0.000 description 4
- 235000009048 phenolic acids Nutrition 0.000 description 4
- XUIMIQQOPSSXEZ-UHFFFAOYSA-N Silicon Chemical compound [Si] XUIMIQQOPSSXEZ-UHFFFAOYSA-N 0.000 description 3
- 238000000855 fermentation Methods 0.000 description 3
- 230000004151 fermentation Effects 0.000 description 3
- 230000004720 fertilization Effects 0.000 description 3
- 229940094952 green tea extract Drugs 0.000 description 3
- 235000020688 green tea extract Nutrition 0.000 description 3
- 238000003306 harvesting Methods 0.000 description 3
- 238000005259 measurement Methods 0.000 description 3
- 230000001737 promoting effect Effects 0.000 description 3
- 239000012488 sample solution Substances 0.000 description 3
- 239000007787 solid Substances 0.000 description 3
- 241001328122 Bacillus clausii Species 0.000 description 2
- VEXZGXHMUGYJMC-UHFFFAOYSA-N Hydrochloric acid Chemical compound Cl VEXZGXHMUGYJMC-UHFFFAOYSA-N 0.000 description 2
- 238000012271 agricultural production Methods 0.000 description 2
- PNEYBMLMFCGWSK-UHFFFAOYSA-N aluminium oxide Inorganic materials [O-2].[O-2].[O-2].[Al+3].[Al+3] PNEYBMLMFCGWSK-UHFFFAOYSA-N 0.000 description 2
- 238000004737 colorimetric analysis Methods 0.000 description 2
- 238000011161 development Methods 0.000 description 2
- 230000007613 environmental effect Effects 0.000 description 2
- 238000001914 filtration Methods 0.000 description 2
- 238000009472 formulation Methods 0.000 description 2
- 238000004128 high performance liquid chromatography Methods 0.000 description 2
- 239000004615 ingredient Substances 0.000 description 2
- 239000002054 inoculum Substances 0.000 description 2
- 238000011835 investigation Methods 0.000 description 2
- 230000003902 lesion Effects 0.000 description 2
- 239000007788 liquid Substances 0.000 description 2
- 230000014759 maintenance of location Effects 0.000 description 2
- 238000003860 storage Methods 0.000 description 2
- 239000000758 substrate Substances 0.000 description 2
- 239000006228 supernatant Substances 0.000 description 2
- USFZMSVCRYTOJT-UHFFFAOYSA-N Ammonium acetate Chemical compound N.CC(O)=O USFZMSVCRYTOJT-UHFFFAOYSA-N 0.000 description 1
- 239000005695 Ammonium acetate Substances 0.000 description 1
- 208000034048 Asymptomatic disease Diseases 0.000 description 1
- 241000193744 Bacillus amyloliquefaciens Species 0.000 description 1
- 241000894006 Bacteria Species 0.000 description 1
- KCXVZYZYPLLWCC-UHFFFAOYSA-N EDTA Chemical compound OC(=O)CN(CC(O)=O)CCN(CC(O)=O)CC(O)=O KCXVZYZYPLLWCC-UHFFFAOYSA-N 0.000 description 1
- 238000007696 Kjeldahl method Methods 0.000 description 1
- ZOKXTWBITQBERF-UHFFFAOYSA-N Molybdenum Chemical compound [Mo] ZOKXTWBITQBERF-UHFFFAOYSA-N 0.000 description 1
- WYWFMUBFNXLFJK-UHFFFAOYSA-N [Mo].[Sb] Chemical compound [Mo].[Sb] WYWFMUBFNXLFJK-UHFFFAOYSA-N 0.000 description 1
- 239000003627 allelochemical Substances 0.000 description 1
- 235000019257 ammonium acetate Nutrition 0.000 description 1
- 229940043376 ammonium acetate Drugs 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 230000000443 biocontrol Effects 0.000 description 1
- 230000015572 biosynthetic process Effects 0.000 description 1
- 238000009264 composting Methods 0.000 description 1
- 244000038559 crop plants Species 0.000 description 1
- 238000001514 detection method Methods 0.000 description 1
- 230000006806 disease prevention Effects 0.000 description 1
- 238000003912 environmental pollution Methods 0.000 description 1
- 238000000605 extraction Methods 0.000 description 1
- 230000035558 fertility Effects 0.000 description 1
- 239000000706 filtrate Substances 0.000 description 1
- 238000009630 liquid culture Methods 0.000 description 1
- 239000007791 liquid phase Substances 0.000 description 1
- 238000004519 manufacturing process Methods 0.000 description 1
- 239000012528 membrane Substances 0.000 description 1
- 239000011785 micronutrient Substances 0.000 description 1
- 235000013369 micronutrients Nutrition 0.000 description 1
- 239000012982 microporous membrane Substances 0.000 description 1
- 239000011733 molybdenum Substances 0.000 description 1
- 229910052750 molybdenum Inorganic materials 0.000 description 1
- 235000015097 nutrients Nutrition 0.000 description 1
- 239000012074 organic phase Substances 0.000 description 1
- 244000052769 pathogen Species 0.000 description 1
- 230000001717 pathogenic effect Effects 0.000 description 1
- 239000000575 pesticide Substances 0.000 description 1
- 239000000447 pesticide residue Substances 0.000 description 1
- 239000012071 phase Substances 0.000 description 1
- 238000005375 photometry Methods 0.000 description 1
- 230000002265 prevention Effects 0.000 description 1
- 239000008213 purified water Substances 0.000 description 1
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- 230000005070 ripening Effects 0.000 description 1
- 238000007789 sealing Methods 0.000 description 1
- 239000002002 slurry Substances 0.000 description 1
- FQENQNTWSFEDLI-UHFFFAOYSA-J sodium diphosphate Chemical compound [Na+].[Na+].[Na+].[Na+].[O-]P([O-])(=O)OP([O-])([O-])=O FQENQNTWSFEDLI-UHFFFAOYSA-J 0.000 description 1
- 229940048086 sodium pyrophosphate Drugs 0.000 description 1
- 238000002798 spectrophotometry method Methods 0.000 description 1
- 239000004575 stone Substances 0.000 description 1
- 238000006467 substitution reaction Methods 0.000 description 1
- 239000000725 suspension Substances 0.000 description 1
- 208000024891 symptom Diseases 0.000 description 1
- 235000019818 tetrasodium diphosphate Nutrition 0.000 description 1
- 239000001577 tetrasodium phosphonato phosphate Substances 0.000 description 1
- 238000009210 therapy by ultrasound Methods 0.000 description 1
- 238000004448 titration Methods 0.000 description 1
- 238000000825 ultraviolet detection Methods 0.000 description 1
- 235000013311 vegetables Nutrition 0.000 description 1
- 238000005303 weighing Methods 0.000 description 1
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- C05—FERTILISERS; MANUFACTURE THEREOF
- C05F—ORGANIC FERTILISERS NOT COVERED BY SUBCLASSES C05B, C05C, e.g. FERTILISERS FROM WASTE OR REFUSE
- C05F3/00—Fertilisers from human or animal excrements, e.g. manure
-
- 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
- A01N61/00—Biocides, pest repellants or attractants, or plant growth regulators containing substances of unknown or undetermined composition, e.g. substances characterised only by the mode of action
-
- 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
- A01N65/00—Biocides, pest repellants or attractants, or plant growth regulators containing material from algae, lichens, bryophyta, multi-cellular fungi or plants, or extracts thereof
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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
- A01N65/00—Biocides, pest repellants or attractants, or plant growth regulators containing material from algae, lichens, bryophyta, multi-cellular fungi or plants, or extracts thereof
- A01N65/08—Magnoliopsida [dicotyledons]
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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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- C05F17/00—Preparation of fertilisers characterised by biological or biochemical treatment steps, e.g. composting or fermentation
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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
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- C12R2001/00—Microorganisms ; Processes using microorganisms
- C12R2001/01—Bacteria or Actinomycetales ; using bacteria or Actinomycetales
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Abstract
The invention relates to a microbial composition for preventing and treating fusarium root rot and application thereof, wherein the composition comprises the following components: a) Bacillus belicus and bacillus siamensis, b) zeolite powder, decomposed cow dung, humic acid, ascorbic acid and tea extract; and wherein the preservation number of bacillus belicus is CCTCC NO: m2022923, the preservation number of the Siamese bacillus is CCTCC NO: m2022925. The composition provided by the invention has high-efficiency antagonism on germs causing root rot of passion fruits, oranges, carnation and tea trees; can be applied to soil to effectively regulate the microbial diversity of the soil and degrade plant rhizosphere phenolic acid substances.
Description
Technical Field
The invention belongs to the technical field of organic fertilizers, and particularly relates to a microbial composition for preventing and treating fusarium root rot and application thereof.
Background
The improvement of crop yield and how to effectively prevent and control soil-borne diseases are the problems to be solved in modern agricultural production, and the scientific, green and sustainable crop cultivation management and control mode has positive significance for a plurality of aspects such as increasing income of farmers, food safety, environmental protection and the like. In modern agricultural production, the purposes of increasing yield and preventing and controlling diseases are usually achieved by using pesticides and fertilizers on a large scale, but at the same time, a large amount of soil environmental problems are brought, such as soil fertility reduction, nutrient loss, environmental pollution and the like, so that the yield of crops is reduced to the quality and vicious circle is carried out for a long time, on the other hand, along with the development of economy, the food safety concept of consumers is gradually enhanced, the requirements on food safety are also higher and higher, and the problems of pesticide residues, chemical fertilizer ripening and the like are more and more concerned. Therefore, there is a need to develop more efficient, environment-friendly, green and safe yield-increasing and disease-preventing products for vegetable production, thereby meeting the great demands of the market for green foods and eliminating the concerns of consumers about food safety hazards.
Disclosure of Invention
In order to overcome the problems in the background art, the invention provides a microbial composition for preventing and treating fusarium root rot and application thereof.
To achieve the above object, in one aspect, the present invention provides a microbial composition for controlling fusarium root rot, comprising: a) Bacillus belicus and Bacillus siamensis,
b) Zeolite powder, decomposed cow dung, humic acid, ascorbic acid and tea extract;
wherein, the preservation number of bacillus bailii is CCTCC NO: m2022923, the preservation number of the Siamese bacillus is CCTCC NO: m2022925.
Further, the weight ratio of the zeolite powder to the decomposed cow dung to the humic acid to the ascorbic acid to the tea extract is 10-20:10-30:5-15:1-5:0.5-1.5.
Further, the water content in the zeolite powder is 3-12 wt%;
and/or, the water content in the decomposed cow dung is 10-15 wt%;
and/or, the humic acid content in the humic acid source is 40-55 wt%;
and/or, the ascorbic acid has an ascorbic acid content of 90 to 99 wt%;
and/or, the content of catechin gallate in the tea extract is 90-98 wt%.
Further, in the zeolite powder, the content of silicon dioxide is 40-60 wt%, the content of aluminum oxide is 8-14 wt%, the content of soluble silicon is 4-13 wt%, the content of soluble aluminum is 2-8 wt%, and the content of main mineral components is 1-5% based on dry matter.
Further, in the decomposed cow dung, the organic matter content is 10-20 wt%, the humic acid content is 0.1-0.5 wt%, the nitrogen content is 0.1-0.5 wt%, the phosphorus content is 0.1-0.5 wt%, and the potassium content is 0.1-0.5 wt% based on dry matter.
Further, the Bacillus bailii is present in an amount of 1X 10 based on the total weight of the composition 8 -1×10 10 cfu/g, content of Siamese bacillus is 1×10 8 -1×10 10 cfu/g。
The second aspect of the invention provides the use of a microbial composition for controlling fusarium root rot in the degradation of soil phenolic acids and/or for modulating the diversity of soil microorganisms and/or for controlling crop diseases; wherein the crop is: passion fruit, citrus, carnation and tea tree.
In a third aspect, the present invention provides a method for modulating microbial diversity in soil and/or controlling crop disease by applying to the soil a microbial composition for controlling fusarium root rot as described above.
Further, the total amount of Bacillus belicus and Bacillus siamensis is not less than 2×10 10 cfu/strain/time; or, the microbial composition for preventing and treating fusarium root rot is applied to the passion fruit planting soil in an amount of 300-400 g/plant/time; the dosage of the fertilizer applied to the carnation planting soil is 200-300 g/plant/time, the dosage of the fertilizer applied to the citrus planting soil is 400-500 g/plant/time, and the dosage of the fertilizer applied to the tea tree planting soil is 300-400 g/plant/time.
Through the technical scheme, the invention has at least the following technical effects: the microbial composition for preventing and treating fusarium root rot provided by the invention has high-efficiency antagonism on germs causing root rot of passion fruits, citrus, carnation and tea trees; can be applied to soil to effectively regulate the microbial diversity of the soil and degrade plant rhizosphere phenolic acid substances.
Detailed Description
Biological preservation in the invention:
the Siamese bacillus (Bacillus siamensis) provided by the invention is preserved in China center for type culture collection (China) for 6 months and 20 days, and has the address of eight paths 299 of Wuchang district of Wuhan, hubei province, and the preservation number of the Wuhan university is CCTCC NO: m2022925.
The bacillus belgium (Bacillus velezensis) provided by the invention is preserved in China center for type culture collection (China) for 6 and 20 days, and has the address of eight paths 299 of Wuchang district of Wuhan, china, university of Wuhan and the preservation number of CCTCC NO: M2022923.
The Siamese bacillus is disclosed in the application number: CN202211189773.X discloses Bacillus bailii as having the following application number: patent document cn202211479280. X.
The inventor finds in the research that the microbial composition for preventing and treating fusarium root rot, which is prepared by mixing zeolite powder, proper amount of decomposed cow dung, humic acid, ascorbic acid and tea extract, and optional bacillus belicus and bacillus siamensis according to a certain proportion, is applied to soil, so that the content of phenolic acid substances in the soil can be reduced, and the allelochemicals of plants can be reduced. In addition, the microbial composition for preventing and treating fusarium root rot can also improve the biological index of soil, regulate the micro-nutrient structure of the soil and promote the formation of the micro-ecological condition of the soil which is beneficial to the healthy growth of crops. The microbial composition for preventing and treating fusarium root rot has good preventing and treating effects on crop diseases, especially on crop soil-borne root rot caused by fusarium.
In one aspect, the invention provides a microbial composition for controlling fusarium root rot, comprising: a) Bacillus beliae and bacillus siamensis, and/or b) zeolite powder, decomposed cow dung, humic acid, ascorbic acid and tea extract; wherein, the preservation number of bacillus bailii is CCTCC NO: m2022923, the preservation number of the Siamese bacillus is CCTCCNO: m2022925.
The formula comprises the following components: the weight ratio of the zeolite powder to the decomposed cow dung to the humic acid to the ascorbic acid to the tea extract is 10-20:10-30:5-15:1-5:0.5-1.5. Preferably 15-20:20-30:5-13:1-4:0.8-1.5. More preferably 18-20:25-30:5-11:1-3:1-1.5.
In the invention, the decomposed cow dung is prepared by decomposing dry cow dung, and the preparation method can comprise the steps of adding sufficient water (controlled at about 40-65%) into the dry cow dung, piling up the dry cow dung, and then sealing and fermenting. When the material turns black brown, the temperature starts to drop to normal temperature, and the fermentation is successful.
In the present invention, the raw materials (such as zeolite powder, decomposed cow dung, humic acid, etc.) used as the microbial inoculum are not particularly limited, and may be any related products which can be used for preparing the microbial inoculum in the field, and may be related products obtained commercially or may be related products prepared by themselves according to the prior art.
The inventors found in the study that the growth promoting effect and disease control effect of the obtained microbial inoculum are better when specific raw materials are adopted. According to a preferred embodiment of the invention, the zeolite powder has a water content of 3 to 12% by weight, preferably 3 to 10% by weight.
Preferably, the zeolite powder has a silica content of 40-60 wt%, an alumina content of 8-14 wt%, a soluble silicon content of 4-13 wt%, a soluble aluminum content of 2-8 wt%, and a main mineral content of 1-5% on a dry matter basis.
More preferably, the zeolite powder has a silica content of 40 to 55 wt.%, an alumina content of 8 to 13 wt.%, a soluble silica content of 4 to 10 wt.%, a soluble aluminum content of 5 to 8 wt.%, and a main mineral component content of 3 to 5% on a dry matter basis.
According to a preferred embodiment of the present invention, wherein the moisture content of the decomposed cow dung is 10-15 wt%, preferably 10-13 wt%.
Preferably, the decomposed cow dung is driedThe organic matter content is 10-20wt%, humic acid content is 0.1-0.5wt%, nitrogen content (calculated as N) is 0.1-0.5wt%, and phosphorus content (calculated as P) 2 O 5 0.1 to 0.5 wt.%, potassium content (in K) 2 O) is 0.1 to 0.5% by weight.
More preferably, the decomposed cow dung contains 12-18 wt% of organic matter, 0.2-0.4 wt% of humic acid, 0.3-0.45 wt% of nitrogen, 0.15-0.25 wt% of phosphorus and 0.1-0.15 wt% of potassium based on dry matter.
According to a preferred embodiment of the present invention, wherein the humic acid content in the source of humic acid is 40-55% by weight;
preferably, the humic acid source has a humic acid content of 50-55% by weight;
more preferably, the humic acid source has a humic acid content of 53-55% by weight;
according to a preferred embodiment of the present invention, wherein the ascorbic acid has an ascorbic acid content of 90 to 99% by weight;
preferably, the ascorbic acid has an ascorbic acid content of 95 to 99 wt%;
more preferably, the ascorbic acid has an ascorbic acid content of 97 to 99 wt%;
according to a preferred embodiment of the present invention, wherein the content of catechin gallate in the green tea extract is 90-98% by weight;
preferably, the content of catechin gallate in the green tea extract is 95-98 wt%;
more preferably, the green tea extract has a catechin gallate content of 97-98% by weight;
in the invention, the content of bacillus behenii and Luo Yabao bacillus is 1 multiplied by 10 based on the total weight of the composition 8 -1×10 10 cfu/g, preferably 1X 10 8 -1×10 9 cfu/g, more preferably 3X 10 8 -8×10 8 cfu/g。
In the present invention, the manner of adding the Bacillus belicus and the Bacillus siamensis is not particularly limited. Any means of adding biocontrol bacteria to the composition in the art may be suitable for use in the present invention. For example, bacillus beliae and bacillus siamensis can be prepared into a microbial inoculum and then added directly into the ingredients according to a certain proportion, or the microbial inoculum and the ingredients are packaged separately and mixed or applied together before use. For example, bacillus beliensis and bacillus siamensis may be subjected to the expansion culture, and the obtained culture solution may be used to prepare the microbial inoculum, for example. In view of the preparation process and convenience in use, it is preferable to add bacillus behenii and bacillus siamensis agents to the aforementioned formulation.
The invention further provides a preparation method of the microbial composition for preventing and treating fusarium root rot, according to a preferred embodiment of the invention, wherein the method comprises the steps of mixing zeolite powder, decomposed cow dung, humic acid, ascorbic acid and tea extract, performing conventional composting to obtain a basic organic fertilizer, adding optional bacillus bailii, bacillus siamensis (or a microbial inoculum thereof) and other raw materials into the basic organic fertilizer according to the above dosage ratio, and uniformly mixing.
In the present invention, the specific storage and use form of the microbial composition for controlling fusarium root rot is not particularly limited. Because the raw materials adopted in the microbial composition for preventing and treating fusarium root rot are basically solid, the preparation method only comprises the steps of uniformly mixing the components according to a proportion. Therefore, the microbial composition for preventing and treating fusarium root rot can be directly stored or used in the form of a solid preparation obtained by preparation. In consideration of different crops or planting modes and planting conditions, the microbial composition for preventing and treating fusarium root rot can be mixed with a certain amount of water to prepare a preparation in the form of liquid or slurry (suspension) for storage or use.
The second aspect of the invention provides the use of a microbial composition for controlling fusarium root rot in the degradation of phenolic acids in soil and/or for regulating the diversity of soil microorganisms and/or for controlling crop diseases; wherein the crop is: passion fruit, citrus, carnation and tea tree.
In the invention, phenolic acid substances are mainly benzoic acid, and the crop diseases are selected from fusarium crop soil-borne root rot, namely, crop root rot caused by fusarium. Preferably fusarium caused crop disease. Such as passion fruit wilt, citrus root rot, carnation root rot, tea tree root rot, etc.
According to some preferred embodiments of the present invention, wherein the pathogen causing soil-borne root rot of the crop is Fusarium oxysporumFusarium oxysporum)。
In a third aspect, the invention provides a method for degrading phenolic acids in soil and/or for regulating microbial diversity in soil and/or for controlling crop diseases, said method comprising applying to the soil a microbial composition for controlling fusarium root rot.
In the present invention, the specific amounts of the microbial compositions for Bacillus belicus and Bacillus siamensis and for controlling fusarium root rot are not particularly limited as long as the effects of promoting crop growth/controlling crop diseases can be achieved.
According to a preferred embodiment of the present invention, wherein the total amount of Bacillus belicus and Bacillus siamensis as described in the above-mentioned mode (1) is not less than 2X 10 10 cfu/strain/time; preferably 1X 10 10 -1×10 11 cfu/strain/time. More preferably 2X 10 10 -8×10 10 cfu/strain/time. Further preferably 3X 10 10 -6×10 10 cfu/strain/time.
According to some preferred embodiments of the present invention, wherein the microbial composition for controlling fusarium root rot according to the above-described modes (2) and (3) is applied to the soil for passion fruit cultivation in an amount of 300 to 400g per plant/time; the application amount of the fertilizer applied to the carnation planting soil is 200-300 g/plant/time, the application amount of the fertilizer applied to the citrus planting soil is 400-500 g/plant/time, and the application amount of the fertilizer applied to the tea tree planting soil is 300-400 g/plant/time. Within the foregoing range: the microbial composition for preventing and treating fusarium root rot is applied to crop planting soil in an amount of 200 g/plant/time, 220 g/plant/time, 250 g/plant/time, 280 g/plant/time, 300 g/plant/time, 320 g/plant/time, 350 g/plant/time, 380 g/plant/time, 500 g/plant/time or any intermediate value between any two of the above values.
In order to make the objects, technical solutions and advantageous effects of the present invention more apparent, preferred embodiments of the present invention will be described in detail below to facilitate understanding by the skilled person.
Examples
The zeolite powder used in this example was purchased from the Shangshou county stone mill run product and the decomposed cow dung was purchased from Jiangsu Dirun organic fertilizer technology Co., ltd, the humic acid was purchased from Shandong Philippine chemical technology Co., ltd, the ascorbic acid was purchased from Jiangxi Lixing biotechnology Co., ltd, and the tea extract was purchased from Shandong jin Shengtai chemical Co., ltd. The microbial inoculum is a liquid culture solution obtained by amplifying Bacillus clausii and Bacillus siamensis (refer to the acquisition method of CN202211189773. X), wherein the content of the Bacillus clausii is 1×10 10 cfu/g, siamese bacillus content of 1×10 10 cfu/ml。
The results of the raw material component measurements for the preparation of the base organic fertilizer are shown in Table 1, wherein the silica content is determined by titration, the aluminum oxide content is determined by EDTA volumetric method, the soluble silicon content is determined by molybdenum blue colorimetric method, the soluble aluminum content is determined by indirect photometry, the organic matter content is determined by ultraviolet spectrophotometry, the humic acid content is determined by sodium pyrophosphate lye extraction method, the nitrogen content (in N) is determined by Kjeldahl method, the phosphorus content (in P 2 O 5 Meter) measured by molybdenum-antimony colorimetric method, potassium content (K) 2 O meter) was measured using a flame photometer. The water content in the detection result is calculated by taking the total weight of the material as a reference, and the rest components are calculated by taking dry matters in the material as a reference.
TABLE 1 basic organic fertilizer Material composition
The microbial agent is prepared according to the formula shown in Table 2, and the specific preparation method comprises the following steps: the raw materials were weighed according to the proportions shown in Table 2, and the microbial agents (solid preparations) were obtained by uniformly mixing the raw materials.
Table 2 microbial composition formulation for controlling fusarium root rot
According to a preferred embodiment of the invention, the bacillus beijerinus and bacillus siamensis inoculants are applied 1-3 times per crop.
In order to obtain better crop growth promotion and disease prevention effects, according to some preferred embodiments of the invention, when the bacillus belicus and the bacillus siamensis microbial inoculum are applied to passion fruits, the bacillus siamensis microbial inoculum is applied once with base fertilizer in spring of the year, and then additional fertilizer is applied 1 time after the growth and development period of plants (5-6 months of the year) (50 plants are planted in each test group).
The carnation plants with consistent growth vigor are selected, the biological microbial inoculum in the embodiment is applied to the rhizosphere of flowers according to the dosage shown in table 2, the base fertilizer is applied once 10-15 days before planting, and the application frequency is 1 time per stubble. Meanwhile, carnation (50 plants are planted in each test group) is planted in farmlands to which the microbial agent (a treatment group) and the microbial agent (a control group) are applied, the disease condition is investigated in a harvesting period, agronomic characters such as plant height and the like of the plants, microbial diversity of rhizosphere soil and the content of phenolic acid (benzoic acid) in the rhizosphere soil of the plants are measured, and the phenolic acid degradation rate is calculated.
The microbial composition 500 g/plant/time for controlling fusarium root rot in table 2 was applied to the rhizosphere of citrus plants (50 plants were planted per test group) such that the microbial agent was used in an amount of not less than 1×10 8 cfu/strain/time; the method comprises the steps of applying for 1 time at intervals of 70-90 days, investigating the control effect of citrus root rot in the citrus harvesting period, measuring agronomic characters such as plant height and the like of plants, microbial diversity of rhizosphere soil and the content of phenolic acid (benzoic acid) in the rhizosphere soil of the plants, and calculating the degradation rate of the phenolic acid.
A batch of potted tea seedlings of substantially uniform initial height (no more than 1cm difference between individual plants) was selected, and 1 plant per pot (50 plants per test group). The microbial agents obtained in the above examples were applied to the rhizosphere soil of tea seedlings in the amounts shown in Table 2, and then cultured simultaneously under other conditions kept identical. The specific fertilization mode comprises the following steps: digging pits of potted tea seedlings before fertilization, namely digging soil deeply along the periphery of the tea seedlings by using a soil digging shovel by 3-5cm and 10-13cm away from the ground, broadcasting microbial inoculum on the periphery of roots of the tea trees, and then covering soil to finish fertilization. Meanwhile, the disease conditions are investigated in the harvest period in the farmland to which the microbial inoculum (treatment group) and the microbial inoculum (control group) provided by the invention are applied, the agronomic characters such as plant height and the like of the plants are measured, the microbial diversity of rhizosphere soil and the content of phenolic acid (benzoic acid) in the rhizosphere soil of the plants are measured, and the phenolic acid degradation rate is calculated.
Passion fruit wilt and carnation wilt investigation and classification standard:
grade 0, healthy root, no disease symptoms;
1 grade, slight damage to root, small amount of disease spots (0-10%);
2, a certain degree of disease spots (11% -25%) are formed on the root;
grade 3, moderate hazard, root lesion spread (26% -50%);
grade 4, serious hazard, most of roots are infected, and only few uninfected tissues (51% -75%);
grade 5, very severe, fully infected roots and even dead plants (76% -100%).
Citrus wilt and tea tree wilt investigation grading criteria:
level 0: asymptomatic condition
Stage 1: the root of the plant has a small amount of disease spots of 0-10%.
3 stages: part of the lesion spots appear in the root 10-25%.
5 stages: root is completely infected.
7 stages: the plants die. The control effect is calculated by adopting the following two formulas:
disease index = [ (Σ (number of disease stages×number of representative stages))/(total number of plants×highest representative stage value) ]×100
Control effect (%) = [ (control group disease index-treatment group disease index)/control group disease index ] ×100%
Determination of the content of phenolic acid (benzoic acid) in the rhizosphere soil of the crop plants:
(1) Benzoic acid standard configuration
0.1g of benzoic acid (analytically pure) is accurately taken and dissolved in purified water, and the volume is fixed to 1L by using a volumetric flask for substitution measurement.
(2) Plant rhizosphere soil sample solution configuration
Respectively weighing each plant rhizosphere soil sample of 25. 25 g, placing the plant rhizosphere soil samples in 500mL conical flasks, respectively adding 200mL of 1mol/L NaOH solution, oscillating, performing ultrasonic treatment for 30 min, and placing the plant rhizosphere soil samples on a shaking table for shaking for 2h. The filtrate was filtered, adjusted p H to 2.5 with hydrochloric acid, allowed to stand overnight and centrifuged. The supernatant was transferred to a 500mL separatory funnel, 200mL ethyl acetate solution was added to the mixture twice, and the organic phase was collected. Recovering ethyl acetate, dissolving the residue with methanol, fixing volume to 5mL, and filtering with 0.45um microporous membrane to obtain sample solution.
(3) Sample measurement
And respectively taking a benzoic acid standard substance and a plant rhizosphere soil sample solution, filtering, and detecting the substrate amount by using HPLC (high performance liquid chromatography), thereby calculating the substrate degradation efficiency. Conditions of high performance liquid phase: taking a sample of the liquid to be detected, adding the sample into a 50mL centrifuge tube, centrifuging for 10min at 8000r/min at 4 ℃, taking a supernatant, taking 1mL of the centrifuged fermentation liquor, and taking the fermentation liquor as a sample, wherein the sample is taken by a 0.22 mu m microporous filter membrane, and a chromatographic column is an Agilent C18 column (250 mm multiplied by 4.0mm multiplied by 5 mu m); the column temperature is 35 ℃; the ultraviolet detection wavelength is 230nm; the mobile phase is methanol: 0.02mol/L ammonium acetate = 5:95; the flow rate is 1.0mL/min; the sample volume was 10. Mu.L, eluted isocratically, peaks were detected and retention time was determined. The benzoic acid standard was determined to have a peak area of 4335.28369 and a retention time of 3.495min.
As can be seen from the data in tables 3, 4 and 5, the prevention and control effect on the fusarium root rot of crops is significantly higher than that of the control when the microbial composition for preventing and controlling the fusarium root rot is applied; the improvement of agronomic characters of crops and the improvement of microbial diversity of rhizosphere soil of crops are also stronger than those of a control; in addition, the microbial composition for preventing and treating fusarium root rot has a stronger degradation effect on the content of the autotoxic substances in the rhizosphere soil of crops than a control.
TABLE 3 test conditions and results of test of control effect of microbial inoculants on Fusarium wilt of crops
* The control group was a blank control group to which no fertilizer was applied, and the fertilizer group was a control group to which the chemical fertilizer of component 3 in example 1 was applied.
TABLE 4 effect of microbial compositions for controlling Fusarium root rot on the growth promoting effect of crops and on the diversity of microbial communities in rhizosphere soil of crops
* The control group was a blank control group to which no fertilizer was applied, and the fertilizer group was a control group to which the chemical fertilizer of component 3 in example 1 was applied.
TABLE 5 degradation of crop rhizosphere phenolic acids by microbial compositions for controlling Fusarium root rot
* The control group was a blank control group to which no fertilizer was applied, and the fertilizer group was a control group to which the chemical fertilizer of component 3 in example 1 was applied.
Finally, it is noted that the above-mentioned preferred embodiments illustrate rather than limit the invention, and that, although the invention has been described in detail with reference to the above-mentioned preferred embodiments, it will be understood by those skilled in the art that various changes in form and details may be made therein without departing from the scope of the invention as defined by the appended claims.
Claims (9)
1. A microbial composition for controlling fusarium root rot, comprising: a) Bacillus beleiensis and bacillus siamensis;
b) Zeolite powder, decomposed cow dung, humic acid, ascorbic acid and tea extract;
wherein, the preservation number of bacillus bailii is CCTCC NO: m2022923, the preservation number of the Siamese bacillus is CCTCC NO: m2022925.
2. The microbial composition for controlling fusarium root rot of claim 1, wherein: the weight ratio of the zeolite powder to the decomposed cow dung to the humic acid to the ascorbic acid to the tea extract is 10-20:10-30:5-15:1-5:0.5-1.5.
3. The microbial composition for controlling fusarium root rot according to claim 2, wherein:
the water content in the zeolite powder is 3-12 wt%;
and/or, the water content in the decomposed cow dung is 10-15 wt%;
and/or, the humic acid content in the humic acid source is 40-55 wt%;
and/or, the ascorbic acid has an ascorbic acid content of 90 to 99 wt%;
and/or, the content of catechin gallate in the tea extract is 90-98 wt%.
4. A microbial composition for controlling fusarium root rot according to claim 3, wherein: in the zeolite powder, the content of silicon dioxide is 40-60 wt%, the content of aluminum oxide is 8-14 wt%, the content of soluble silicon is 4-13 wt%, the content of soluble aluminum is 2-8 wt%, and the content of main mineral components is 1-5%.
5. A microbial composition for controlling fusarium root rot according to claim 3, wherein: in the decomposed cow dung, the organic matter content is 10-20 wt%, the humic acid content is 0.1-0.5 wt%, the nitrogen content is 0.1-0.5 wt%, the phosphorus content is 0.1-0.5 wt%, and the potassium content is 0.1-0.5 wt% based on dry matter.
6. The microbial composition for controlling fusarium root rot of claim 5, wherein: the content of Bacillus bailii is 1×10 based on the total weight of the composition 8 -1×10 10 cfu/g, content of Siamese bacillus is 1×10 8 -1×10 10 cfu/g。
7. Use of a microbial composition for controlling fusarium root rot according to any one of claims 1-6, characterized in that: the application of the microbial composition for preventing and treating fusarium root rot in degrading soil phenolic acid and/or regulating the diversity of soil microorganisms and/or preventing and treating crop diseases; wherein the crops are: passion fruit, citrus, carnation or tea tree.
8. The use of a microbial composition for controlling fusarium root rot according to claim 7, characterized in that: the microbial composition for preventing and treating fusarium root rot is applied to soil.
9. The use of a microbial composition for controlling fusarium root rot according to claim 8, wherein the total amount of bacillus belicus and bacillus siamensis is not less than 2 x 10 10 cfu/strain/time; alternatively, the microbial composition is applied to the passion fruit planting soil in an amount of 300-400 g/plant/time; the application amount of the fertilizer applied to the carnation planting soil is 200-300 g/plant/time, the application amount of the fertilizer applied to the citrus planting soil is 400-500 g/plant/time, and the application amount of the fertilizer applied to the tea tree planting soil is 300-400 g/plant/time.
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申云鑫等: "三株具生防功能芽孢杆菌的分离鉴定及其生物活性研究", 生物技术通报, pages 267 - 277 * |
申云鑫等: "贝莱斯芽胞杆菌SH-1471可湿性粉剂研制及其对番茄枯萎病的防治效果", 中国生物防治学报, pages 904 - 914 * |
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