CN114903055B - Wettable powder for preventing and treating black spot of celery cabbage, preparation method and application - Google Patents
Wettable powder for preventing and treating black spot of celery cabbage, preparation method and application Download PDFInfo
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- 206010027146 Melanoderma Diseases 0.000 title claims abstract description 57
- 239000004563 wettable powder Substances 0.000 title claims abstract description 41
- 238000002360 preparation method Methods 0.000 title claims abstract description 18
- 241000499436 Brassica rapa subsp. pekinensis Species 0.000 title claims description 13
- 235000010149 Brassica rapa subsp chinensis Nutrition 0.000 title claims description 12
- 235000000536 Brassica rapa subsp pekinensis Nutrition 0.000 title claims description 12
- 241000194107 Bacillus megaterium Species 0.000 claims abstract description 73
- 101100471831 Neurospora crassa (strain ATCC 24698 / 74-OR23-1A / CBS 708.71 / DSM 1257 / FGSC 987) rpl-5 gene Proteins 0.000 claims abstract description 66
- 235000011301 Brassica oleracea var capitata Nutrition 0.000 claims abstract description 46
- 240000007124 Brassica oleracea Species 0.000 claims abstract description 45
- 235000003899 Brassica oleracea var acephala Nutrition 0.000 claims abstract description 45
- 235000001169 Brassica oleracea var oleracea Nutrition 0.000 claims abstract description 45
- 239000000843 powder Substances 0.000 claims abstract description 35
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 claims abstract description 34
- 230000001580 bacterial effect Effects 0.000 claims abstract description 29
- 201000010099 disease Diseases 0.000 claims abstract description 28
- 239000007788 liquid Substances 0.000 claims abstract description 23
- 239000003223 protective agent Substances 0.000 claims abstract description 22
- 239000002270 dispersing agent Substances 0.000 claims abstract description 19
- 239000004094 surface-active agent Substances 0.000 claims abstract description 9
- 238000000855 fermentation Methods 0.000 claims description 20
- 230000004151 fermentation Effects 0.000 claims description 20
- VYPSYNLAJGMNEJ-UHFFFAOYSA-N Silicium dioxide Chemical group O=[Si]=O VYPSYNLAJGMNEJ-UHFFFAOYSA-N 0.000 claims description 11
- 239000002609 medium Substances 0.000 claims description 11
- 241001052560 Thallis Species 0.000 claims description 9
- YDEXUEFDPVHGHE-GGMCWBHBSA-L disodium;(2r)-3-(2-hydroxy-3-methoxyphenyl)-2-[2-methoxy-4-(3-sulfonatopropyl)phenoxy]propane-1-sulfonate Chemical group [Na+].[Na+].COC1=CC=CC(C[C@H](CS([O-])(=O)=O)OC=2C(=CC(CCCS([O-])(=O)=O)=CC=2)OC)=C1O YDEXUEFDPVHGHE-GGMCWBHBSA-L 0.000 claims description 9
- DBMJMQXJHONAFJ-UHFFFAOYSA-M Sodium laurylsulphate Chemical compound [Na+].CCCCCCCCCCCCOS([O-])(=O)=O DBMJMQXJHONAFJ-UHFFFAOYSA-M 0.000 claims description 8
- DPXJVFZANSGRMM-UHFFFAOYSA-N acetic acid;2,3,4,5,6-pentahydroxyhexanal;sodium Chemical compound [Na].CC(O)=O.OCC(O)C(O)C(O)C(O)C=O DPXJVFZANSGRMM-UHFFFAOYSA-N 0.000 claims description 8
- 239000001768 carboxy methyl cellulose Substances 0.000 claims description 8
- 235000019812 sodium carboxymethyl cellulose Nutrition 0.000 claims description 8
- 229920001027 sodium carboxymethylcellulose Polymers 0.000 claims description 8
- 238000012258 culturing Methods 0.000 claims description 7
- 229920002472 Starch Polymers 0.000 claims description 6
- 230000003213 activating effect Effects 0.000 claims description 6
- -1 alkyl naphthalene sulfonate Chemical compound 0.000 claims description 6
- 239000007787 solid Substances 0.000 claims description 6
- 239000008107 starch Substances 0.000 claims description 6
- 235000019698 starch Nutrition 0.000 claims description 6
- 238000001035 drying Methods 0.000 claims description 5
- 238000004321 preservation Methods 0.000 claims description 5
- 240000007472 Leucaena leucocephala Species 0.000 claims description 4
- 235000010643 Leucaena leucocephala Nutrition 0.000 claims description 4
- 230000004913 activation Effects 0.000 claims description 3
- 239000000463 material Substances 0.000 claims description 3
- 238000000034 method Methods 0.000 claims description 3
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- 240000000103 Potentilla erecta Species 0.000 claims description 2
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- 238000005119 centrifugation Methods 0.000 claims description 2
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- 238000009629 microbiological culture Methods 0.000 claims description 2
- 238000011282 treatment Methods 0.000 abstract description 30
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- 239000000126 substance Substances 0.000 abstract description 6
- 239000000203 mixture Substances 0.000 abstract description 5
- 239000000575 pesticide Substances 0.000 abstract description 5
- 239000000447 pesticide residue Substances 0.000 abstract description 4
- 244000052616 bacterial pathogen Species 0.000 abstract description 3
- 241000894006 Bacteria Species 0.000 abstract description 2
- 239000003795 chemical substances by application Substances 0.000 abstract description 2
- 230000000813 microbial effect Effects 0.000 abstract description 2
- VTYYLEPIZMXCLO-UHFFFAOYSA-L Calcium carbonate Chemical compound [Ca+2].[O-]C([O-])=O VTYYLEPIZMXCLO-UHFFFAOYSA-L 0.000 description 6
- 241000196324 Embryophyta Species 0.000 description 6
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 description 6
- HEMHJVSKTPXQMS-UHFFFAOYSA-M Sodium hydroxide Chemical compound [OH-].[Na+] HEMHJVSKTPXQMS-UHFFFAOYSA-M 0.000 description 6
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 6
- 238000012216 screening Methods 0.000 description 5
- 239000000243 solution Substances 0.000 description 5
- 235000010489 acacia gum Nutrition 0.000 description 4
- 235000020183 skimmed milk Nutrition 0.000 description 4
- 238000009736 wetting Methods 0.000 description 4
- 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 description 3
- 239000001888 Peptone Substances 0.000 description 3
- 108010080698 Peptones Proteins 0.000 description 3
- 239000001785 acacia senegal l. willd gum Substances 0.000 description 3
- 239000012752 auxiliary agent Substances 0.000 description 3
- 230000000443 biocontrol Effects 0.000 description 3
- 229910000019 calcium carbonate Inorganic materials 0.000 description 3
- 239000011248 coating agent Substances 0.000 description 3
- 238000000576 coating method Methods 0.000 description 3
- 238000007865 diluting Methods 0.000 description 3
- 239000004021 humic acid Substances 0.000 description 3
- 235000019319 peptone Nutrition 0.000 description 3
- 238000011160 research Methods 0.000 description 3
- 238000004062 sedimentation Methods 0.000 description 3
- 239000011780 sodium chloride Substances 0.000 description 3
- 235000019832 sodium triphosphate Nutrition 0.000 description 3
- 235000013311 vegetables Nutrition 0.000 description 3
- LWEAHXKXKDCSIE-UHFFFAOYSA-M 2,3-di(propan-2-yl)naphthalene-1-sulfonate Chemical compound C1=CC=C2C(S([O-])(=O)=O)=C(C(C)C)C(C(C)C)=CC2=C1 LWEAHXKXKDCSIE-UHFFFAOYSA-M 0.000 description 2
- 239000005995 Aluminium silicate Substances 0.000 description 2
- VEXZGXHMUGYJMC-UHFFFAOYSA-N Hydrochloric acid Chemical compound Cl VEXZGXHMUGYJMC-UHFFFAOYSA-N 0.000 description 2
- DGAQECJNVWCQMB-PUAWFVPOSA-M Ilexoside XXIX Chemical compound C[C@@H]1CC[C@@]2(CC[C@@]3(C(=CC[C@H]4[C@]3(CC[C@@H]5[C@@]4(CC[C@@H](C5(C)C)OS(=O)(=O)[O-])C)C)[C@@H]2[C@]1(C)O)C)C(=O)O[C@H]6[C@@H]([C@H]([C@@H]([C@H](O6)CO)O)O)O.[Na+] DGAQECJNVWCQMB-PUAWFVPOSA-M 0.000 description 2
- CSNNHWWHGAXBCP-UHFFFAOYSA-L Magnesium sulfate Chemical compound [Mg+2].[O-][S+2]([O-])([O-])[O-] CSNNHWWHGAXBCP-UHFFFAOYSA-L 0.000 description 2
- 241001465754 Metazoa Species 0.000 description 2
- 125000000217 alkyl group Chemical group 0.000 description 2
- 235000012211 aluminium silicate Nutrition 0.000 description 2
- 230000000721 bacterilogical effect Effects 0.000 description 2
- 229940041514 candida albicans extract Drugs 0.000 description 2
- 239000006185 dispersion Substances 0.000 description 2
- 238000011835 investigation Methods 0.000 description 2
- NLYAJNPCOHFWQQ-UHFFFAOYSA-N kaolin Chemical compound O.O.O=[Al]O[Si](=O)O[Si](=O)O[Al]=O NLYAJNPCOHFWQQ-UHFFFAOYSA-N 0.000 description 2
- 239000002068 microbial inoculum Substances 0.000 description 2
- PSZYNBSKGUBXEH-UHFFFAOYSA-M naphthalene-1-sulfonate Chemical compound C1=CC=C2C(S(=O)(=O)[O-])=CC=CC2=C1 PSZYNBSKGUBXEH-UHFFFAOYSA-M 0.000 description 2
- LWIHDJKSTIGBAC-UHFFFAOYSA-K potassium phosphate Substances [K+].[K+].[K+].[O-]P([O-])([O-])=O LWIHDJKSTIGBAC-UHFFFAOYSA-K 0.000 description 2
- 230000002265 prevention Effects 0.000 description 2
- 230000002829 reductive effect Effects 0.000 description 2
- 239000011734 sodium Substances 0.000 description 2
- 229910052708 sodium Inorganic materials 0.000 description 2
- 238000005507 spraying Methods 0.000 description 2
- 239000000725 suspension Substances 0.000 description 2
- 239000012138 yeast extract Substances 0.000 description 2
- FRASJONUBLZVQX-UHFFFAOYSA-N 1,4-naphthoquinone Chemical compound C1=CC=C2C(=O)C=CC(=O)C2=C1 FRASJONUBLZVQX-UHFFFAOYSA-N 0.000 description 1
- 229920001817 Agar Polymers 0.000 description 1
- 241001149961 Alternaria brassicae Species 0.000 description 1
- 235000011299 Brassica oleracea var botrytis Nutrition 0.000 description 1
- 240000003259 Brassica oleracea var. botrytis Species 0.000 description 1
- 244000178937 Brassica oleracea var. capitata Species 0.000 description 1
- 241000219193 Brassicaceae Species 0.000 description 1
- 239000005747 Chlorothalonil Substances 0.000 description 1
- 206010059866 Drug resistance Diseases 0.000 description 1
- 235000019733 Fish meal Nutrition 0.000 description 1
- 244000068988 Glycine max Species 0.000 description 1
- 235000010469 Glycine max Nutrition 0.000 description 1
- 229920000084 Gum arabic Polymers 0.000 description 1
- 239000005802 Mancozeb Substances 0.000 description 1
- 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 description 1
- 229930006000 Sucrose Natural products 0.000 description 1
- 239000005842 Thiophanate-methyl Substances 0.000 description 1
- 240000008042 Zea mays Species 0.000 description 1
- 235000005824 Zea mays ssp. parviglumis Nutrition 0.000 description 1
- 235000002017 Zea mays subsp mays Nutrition 0.000 description 1
- 239000000205 acacia gum Substances 0.000 description 1
- 239000008272 agar Substances 0.000 description 1
- BFNBIHQBYMNNAN-UHFFFAOYSA-N ammonium sulfate Chemical compound N.N.OS(O)(=O)=O BFNBIHQBYMNNAN-UHFFFAOYSA-N 0.000 description 1
- 229910052921 ammonium sulfate Inorganic materials 0.000 description 1
- 235000011130 ammonium sulphate Nutrition 0.000 description 1
- 230000000844 anti-bacterial effect Effects 0.000 description 1
- 230000001174 ascending effect Effects 0.000 description 1
- 239000003899 bactericide agent Substances 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 230000004071 biological effect Effects 0.000 description 1
- CRQQGFGUEAVUIL-UHFFFAOYSA-N chlorothalonil Chemical compound ClC1=C(Cl)C(C#N)=C(Cl)C(C#N)=C1Cl CRQQGFGUEAVUIL-UHFFFAOYSA-N 0.000 description 1
- 238000013329 compounding Methods 0.000 description 1
- 235000005822 corn Nutrition 0.000 description 1
- 239000013530 defoamer Substances 0.000 description 1
- 238000009792 diffusion process Methods 0.000 description 1
- 238000010790 dilution Methods 0.000 description 1
- 239000012895 dilution Substances 0.000 description 1
- ZPWVASYFFYYZEW-UHFFFAOYSA-L dipotassium hydrogen phosphate Chemical compound [K+].[K+].OP([O-])([O-])=O ZPWVASYFFYYZEW-UHFFFAOYSA-L 0.000 description 1
- 235000019797 dipotassium phosphate Nutrition 0.000 description 1
- 229910000396 dipotassium phosphate Inorganic materials 0.000 description 1
- 239000012153 distilled water Substances 0.000 description 1
- 238000002474 experimental method Methods 0.000 description 1
- 239000004467 fishmeal Substances 0.000 description 1
- 235000013312 flour Nutrition 0.000 description 1
- 238000009472 formulation Methods 0.000 description 1
- 230000035784 germination Effects 0.000 description 1
- 230000002401 inhibitory effect Effects 0.000 description 1
- 230000003902 lesion Effects 0.000 description 1
- 230000007774 longterm Effects 0.000 description 1
- 229910052943 magnesium sulfate Inorganic materials 0.000 description 1
- 235000019341 magnesium sulphate Nutrition 0.000 description 1
- 229940099596 manganese sulfate Drugs 0.000 description 1
- 239000011702 manganese sulphate Substances 0.000 description 1
- 235000007079 manganese sulphate Nutrition 0.000 description 1
- SQQMAOCOWKFBNP-UHFFFAOYSA-L manganese(II) sulfate Chemical compound [Mn+2].[O-]S([O-])(=O)=O SQQMAOCOWKFBNP-UHFFFAOYSA-L 0.000 description 1
- 238000004519 manufacturing process Methods 0.000 description 1
- 235000012054 meals Nutrition 0.000 description 1
- 230000002906 microbiologic effect Effects 0.000 description 1
- 244000005700 microbiome Species 0.000 description 1
- 238000002156 mixing Methods 0.000 description 1
- 229910000402 monopotassium phosphate Inorganic materials 0.000 description 1
- 235000019796 monopotassium phosphate Nutrition 0.000 description 1
- 230000000877 morphologic effect Effects 0.000 description 1
- 231100000989 no adverse effect Toxicity 0.000 description 1
- GNSKLFRGEWLPPA-UHFFFAOYSA-M potassium dihydrogen phosphate Chemical compound [K+].OP(O)([O-])=O GNSKLFRGEWLPPA-UHFFFAOYSA-M 0.000 description 1
- 238000012827 research and development Methods 0.000 description 1
- 238000006467 substitution reaction Methods 0.000 description 1
- 239000005720 sucrose Substances 0.000 description 1
- 238000012360 testing method Methods 0.000 description 1
- QGHREAKMXXNCOA-UHFFFAOYSA-N thiophanate-methyl Chemical compound COC(=O)NC(=S)NC1=CC=CC=C1NC(=S)NC(=O)OC QGHREAKMXXNCOA-UHFFFAOYSA-N 0.000 description 1
Classifications
-
- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01N—PRESERVATION OF BODIES OF HUMANS OR ANIMALS OR PLANTS OR PARTS THEREOF; BIOCIDES, e.g. AS DISINFECTANTS, AS PESTICIDES OR AS HERBICIDES; PEST REPELLANTS OR ATTRACTANTS; PLANT GROWTH REGULATORS
- A01N63/00—Biocides, pest repellants or attractants, or plant growth regulators containing microorganisms, viruses, microbial fungi, animals or substances produced by, or obtained from, microorganisms, viruses, microbial fungi or animals, e.g. enzymes or fermentates
- A01N63/20—Bacteria; Substances produced thereby or obtained therefrom
- A01N63/22—Bacillus
-
- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01N—PRESERVATION OF BODIES OF HUMANS OR ANIMALS OR PLANTS OR PARTS THEREOF; BIOCIDES, e.g. AS DISINFECTANTS, AS PESTICIDES OR AS HERBICIDES; PEST REPELLANTS OR ATTRACTANTS; PLANT GROWTH REGULATORS
- A01N25/00—Biocides, pest repellants or attractants, or plant growth regulators, characterised by their forms, or by their non-active ingredients or by their methods of application, e.g. seed treatment or sequential application; Substances for reducing the noxious effect of the active ingredients to organisms other than pests
- A01N25/12—Powders or granules
- A01N25/14—Powders or granules wettable
-
- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02A—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE
- Y02A50/00—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE in human health protection, e.g. against extreme weather
- Y02A50/30—Against vector-borne diseases, e.g. mosquito-borne, fly-borne, tick-borne or waterborne diseases whose impact is exacerbated by climate change
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- Life Sciences & Earth Sciences (AREA)
- General Health & Medical Sciences (AREA)
- Health & Medical Sciences (AREA)
- Engineering & Computer Science (AREA)
- Zoology (AREA)
- Wood Science & Technology (AREA)
- Pest Control & Pesticides (AREA)
- Plant Pathology (AREA)
- Dentistry (AREA)
- Agronomy & Crop Science (AREA)
- Environmental Sciences (AREA)
- Microbiology (AREA)
- Virology (AREA)
- Biotechnology (AREA)
- Toxicology (AREA)
- Agricultural Chemicals And Associated Chemicals (AREA)
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
Abstract
The invention discloses wettable powder for preventing and treating cabbage black spot, a preparation method and application thereof, relates to the field of microbial agents, and solves the problems of pesticide residue and environmental pollution existing in the prior art that chemical pesticides are adopted for preventing and treating cabbage black spot. The wettable powder for preventing and treating cabbage black spot comprises the following components in parts by weight: 25 to 65 parts of bacillus megaterium CRP-4 bacterial powder, 15 to 25 parts of dispersing agent, 5 to 15 parts of surfactant, 1 to 6 parts of protective agent and 10 to 80 parts of carrier. The wettable powder for preventing and treating cabbage black spot is used before or at the beginning of the disease of cabbage black spot, and can protect non-disease crops from being infected by black spot bacteria; for the crops with disease, the composition has good treatment effect and can obviously prevent and treat the spread of germs; the wettable powder for preventing and treating cabbage black spot is diluted 1000-2000 times and sprayed on leaf surfaces, and bacterial liquid can be completely absorbed by the leaf surfaces without leaving any surface residues and causing environmental pollution.
Description
Technical Field
The invention relates to the technical field of microbial agents, in particular to wettable powder for preventing and treating cabbage black spot, a preparation method and application thereof.
Background
The black spot disease of the Chinese cabbage is a disease caused by alternaria brassicae and the like and occurring in the Chinese cabbage. The black spot disease of the Chinese cabbage mainly occurs on the outer leaves or outer leaf bulbs of the Chinese cabbage, and cotyledons, petioles, pedicel and seed pods can be possibly damaged. Leaf onset is manifested as: small black spots are generated on the leaves, the disease spots are gradually enlarged to form black brown concentric circular patterns, yellow halation is sometimes arranged around the disease spots, and the diameter of the disease spots is 2-6 mm; or from the edge of the leaf, brown to blackish brown concentric rings are formed, and later multiple lesions merge, sometimes with intermediate perforations or breaks. The lower leaf of the plant is frequently seriously ill, and then gradually develops to the upper leaf until the whole plant dies.
Cabbage is a cruciferae and cabbage vegetable black spot disease is common in China. In recent years, the incidence rate of cabbage black spot disease is in an ascending trend in Gansu, and the disease-resistant variety is also in the disease-resistant condition. The disease only damages cruciferous vegetables, more cabbage, cabbage and cauliflower occur, the quality of the cruciferous vegetables is reduced after the appearance of leaf bulbs and flower bulbs is stained except that the yield is reduced when the disease is serious, and the seed strain is infected, so that the seed yield is affected, and the seeds can be brought into charge; however, when the cabbage is damaged, the leaves become bitter, and eating is affected.
At present, in the aspect of preventing and treating the black spot, a large amount of chemical, such as 70% mancozeb wettable powder, 75% chlorothalonil wettable powder and the like, is used for preventing and treating the black spot. The chemical pesticide has a certain control effect on cabbage black spot control, but with long-term use of the chemical pesticide, the problems of pesticide residue and environmental pollution are more serious. Therefore, research and development of new biocontrol bactericides for preventing and treating cabbage black spot is widely focused. However, the research on preventing and controlling cabbage black spot by using the biocontrol microbial inoculum is very few at present, so that the research on preventing and controlling cabbage black spot by using the preparation form of the biocontrol microbial inoculum has important research significance.
Disclosure of Invention
The invention aims to provide wettable powder for preventing and treating cabbage black spot, a preparation method and application thereof, and solves the technical problems of pesticide residue and environmental pollution existing in the prior art that chemical pesticides are adopted for preventing and treating cabbage black spot. The technical effects that can be produced by the preferred technical scheme of the present invention are described in detail below.
In order to achieve the above purpose, the present invention provides the following technical solutions:
the invention relates to wettable powder for preventing and treating cabbage black spot, which comprises the following components in parts by weight: 25 to 65 parts of bacillus megaterium CRP-4 bacterial powder, 15 to 25 parts of dispersing agent, 5 to 15 parts of surfactant, 1 to 6 parts of protective agent and 10 to 80 parts of carrier.
The bacillus megatherium (Bacillus megaterium) is provided with a CRP-4 number, a CGMCC No.18458 and a preservation date of 2019, 09 and 02 days, and the preservation unit is as follows: china general microbiological culture Collection center, preservation address: no.1 and No. 3 of the north cinquefoil of the morning sun area of beijing city.
The bacillus megatherium of the invention is CRP-4, which is a strain separated from greenhouse of microorganism fermentation engineering research center, inc. of Yunnan province. The strain has obvious inhibiting effect on the germination of black spot spores indoors. From the comparison and analysis result of the 16SrDNA sequence similarity and the phylogenetic tree, the CRP-4 and the bacillus megatherium (Bacillus megaterium) have the closest relationship, the homology is the highest, the similarity is 99 percent, the CRP-4 and the Bacillus megaterium are positioned on the same branch according to the similarity and the homology analysis of the phylogenetic tree, the evolution relationship and the distance of the CRP-4 and the Bacillus megaterium are the closest, the similarity is 100 percent, and the strain is identified to be the bacillus megatherium by combining morphological characteristics, culture characteristics and physiological and biochemical characteristics.
According to a preferred embodiment, the bacillus megaterium CRP-4 bacterial powder is prepared by taking bacillus megaterium CRP-4 as a bacterial strain, and performing activation, culture, fermentation, centrifugation and drying.
According to a preferred embodiment, the effective viable count in the bacillus megaterium CRP-4 powder is not less than 1000 hundred million/gram.
According to a preferred embodiment, the dispersant comprises one or more of sodium dodecyl sulfate, alkyl naphthalene sulfonate and sodium lignin sulfonate; the surfactant comprises sodium lignin sulfonate; the protective agent comprises one or two of acacia and sodium carboxymethylcellulose, and the carrier comprises one or more of light calcium carbonate, diatomite and kaolin.
According to a preferred embodiment, the dispersant comprises one or more of the following components in parts by weight: 1 to 10 parts of sodium dodecyl sulfate, 10 to 25 parts of alkyl naphthalene sulfonate and 1 to 6 parts of sodium lignin sulfonate.
According to a preferred embodiment, the protective agent comprises one or both of the following components in parts by weight: 1 to 6 parts of Arabic gum and 1 to 5 parts of sodium carboxymethyl cellulose.
According to a preferred embodiment, the carrier comprises one or more of the following components in parts by weight: 20 to 35 parts of light calcium carbonate, 10 to 25 parts of diatomite and 10 to 20 parts of kaolin.
The preparation method of the wettable powder for preventing and treating cabbage black spot according to any one of the technical schemes comprises the following steps:
preparing bacillus megatherium CRP-4 bacterial powder;
25 to 65 parts of bacillus megaterium CRP-4 bacterial powder, 15 to 25 parts of dispersing agent, 5 to 15 parts of surfactant, 1 to 6 parts of protective agent and 10 to 80 parts of carrier are compounded to prepare the wettable powder for preventing and treating the black spot of the celery cabbage.
According to a preferred embodiment, the preparation of bacillus megaterium CRP-4 meal comprises the following steps:
activating strains: activating the bacillus megatherium CRP-4 stored at the temperature of minus 80 ℃ on an LB solid medium for 12 to 16 hours to obtain activated bacillus megatherium CRP-4;
seed liquid preparation: scraping a loop of activated bacillus megatherium CRP-4 by using a sterile inoculating loop, inoculating the bacillus megatherium CRP-4 into 50ml of LB liquid medium, and culturing for 24 hours at the temperature of 35 ℃ and the rotating speed of 160rpm to obtain bacillus megatherium CRP-4 seed liquid;
preparing fermentation liquid: inoculating the bacillus megaterium CRP-4 seed liquid into a workshop fermentation material culture medium according to the volume ratio of the strain to the culture liquid of 1:120, and culturing for 36-48 hours at the temperature of 36.5-37.5 ℃ and the rotating speed of 120rpm/min to obtain bacillus megaterium CRP-4 fermentation liquid, wherein the effective viable count of the bacillus megaterium CRP-4 fermentation liquid is not less than 40 multiplied by 10 8 cfu/g；
Centrifuging and drying: pouring the fermentation liquor of the bacillus megatherium CRP-4 into a centrifugal machine through equipment, preparing centrifugal thalli of the bacillus megatherium CRP-4 under the condition that the rotating speed of the centrifugal machine is 4000rpm/min,
adding soluble starch and diatomite into the centrifugal thalli of the bacillus megaterium CRP-4, and transferring the centrifugal thalli of the bacillus megaterium CRP-4 added with the soluble starch and the diatomite into spray drying equipment to obtain bacillus megaterium CRP-4 bacterial powder.
The application of the wettable powder for preventing and treating cabbage black spot in the invention is that the wettable powder for preventing and treating cabbage black spot is diluted 1000-2000 times before or in the initial stage of the occurrence of cabbage black spot and sprayed on leaf surfaces.
The wettable powder for preventing and treating cabbage black spot, the preparation method and the application provided by the invention have at least the following beneficial technical effects:
the wettable powder for preventing and treating cabbage black spot is prepared by preparing bacillus megaterium CRP-4 bacterial powder, and then compounding bacillus megaterium CRP-4 bacterial powder, a dispersing agent, a surfactant, a protective agent and a carrier in proportion, is used before or at the beginning of the incidence of cabbage black spot in the process of planting crucifer plants, and can protect non-incidence crops from being infected by black spot bacteria; for the crops with the disease, the wettable powder for preventing and treating the black spot disease of the celery cabbage can play a good role in treatment, can remove germs which invade the plant body, and can obviously prevent and treat the spread of the germs; on the other hand, the wettable powder for preventing and treating the black spot of the celery cabbage is diluted by 1000-2000 times and sprayed on leaf surfaces, bacterial liquid can be completely absorbed by the leaf surfaces, no surface residues are left, no environmental pollution is caused, no adverse effect is generated on biological activities such as field animals, natural enemies of the field animals and the like, and the plant is not easy to generate drug resistance.
The wettable powder for preventing and treating cabbage black spot, the preparation method and the application solve the technical problems of pesticide residue and environmental pollution existing in the prior art that chemical pesticides are adopted to prevent and treat cabbage black spot.
Detailed Description
In order to make the objects, technical solutions and advantages of the present invention more apparent, the technical solutions of the present invention will be described in detail below. It will be apparent that the described embodiments are only some, but not all, embodiments of the invention. All other embodiments, based on the examples herein, which are within the scope of the invention as defined by the claims, will be within the scope of the invention as defined by the claims.
The present invention will be described in detail with reference to examples 1 to 5.
Example 1
The preparation process of the bacillus megaterium CRP-4 bacterial powder is described in detail in the embodiment.
The preparation of bacillus megaterium CRP-4 bacterial powder in the embodiment comprises the following steps:
s1: activating strains. Activating the bacillus megatherium CRP-4 stored at the temperature of minus 80 ℃ on an LB solid medium for 12 to 16 hours to obtain the activated bacillus megatherium CRP-4. Preferably, the activation time of the Bacillus megaterium CRP-4 on LB solid medium is, for example, 12h, 14h or 16h.
S2: and (5) preparing seed liquid. Scraping a loop of activated bacillus megatherium CRP-4 by using a sterile inoculating loop, inoculating the bacillus megatherium CRP-4 into 50ml of LB liquid medium, and culturing for 24 hours at the temperature of 35 ℃ and the rotating speed of 160rpm to obtain bacillus megatherium CRP-4 seed liquid.
S3: and (5) preparing fermentation liquor. Inoculating bacillus megaterium CRP-4 seed solution according to the volume ratio of the strain to the culture solution of 1:120Culturing in workshop fermentation medium at 36.5-37.5 deg.c and 120rpm/min for 36-48 hr to obtain bacillus megaterium CRP-4 fermentation liquid with effective viable count not lower than 40×10 8 cfu/g. Preferably, the fermentation temperature is, for example, 36.5 ℃, 37 ℃ or 37.5 ℃. Preferably, the fermentation time is, for example, 36h, 42h or 48h.
S4: centrifuging and drying. Pouring the bacillus megatherium CRP-4 fermentation liquor into a centrifuge through equipment, and preparing centrifugal thalli of bacillus megatherium CRP-4 under the condition that the rotational speed of the centrifuge is 4000 rpm/min; adding soluble starch and diatomite into the centrifugal thalli of the bacillus megaterium CRP-4, transferring the centrifugal thalli of the bacillus megaterium CRP-4 added with the soluble starch and the diatomite into spray drying equipment to obtain bacillus megaterium CRP-4 bacterial powder, wherein the effective viable count in the bacillus megaterium CRP-4 bacterial powder is not less than 1000 hundred million/g.
According to a preferred embodiment, in step S1, the LB solid medium comprises the following components by weight: 10g/L bacteriological peptone, 5g/L yeast extract powder, 10g/L sodium chloride, 15g/L agar powder and 1L water.
According to a preferred embodiment, in step S2, the LB liquid medium comprises the following components by weight: 10g/L bacteriological peptone, 5g/L yeast extract powder, 10g/L sodium chloride and 1L water.
According to a preferred embodiment, in step S3, the fermenter broth comprises the following components by weight: 1.0 to 1.6 percent of soybean powder, 0.1 to 0.5 percent of sucrose, 0.2 to 0.6 percent of peptone, 0.2 to 0.6 percent of corn flour, 0.3 to 0.9 percent of calcium carbonate, 0.1 to 0.3 percent of fish meal, 0.01 to 0.08 percent of ammonium sulfate, 0.02 to 0.05 percent of magnesium sulfate, 0.01 to 0.04 percent of monopotassium phosphate, 0.02 to 0.05 percent of dipotassium phosphate, 0.01 to 0.04 percent of manganese sulfate, 0.01 to 0.04 percent of sodium hydroxide, 0.02 to 0.03 percent of sodium chloride, 0.5 to 1.5 percent of defoamer, and the balance of water, and adjusting the pH value to 6.8 to 7.5 by hydrochloric acid or sodium hydroxide.
Example 2
The present example describes in detail the screening of the dispersing agent in the wettable powder for controlling cabbage black spot according to the present invention.
The bacillus megaterium CRP-4 bacterial powder obtained in the example 1 is mixed with a carrier (such as diatomite), then is mixed with a dispersing agent and dried, 2g of the dispersing agent and 1g of the bacterial powder are respectively and evenly mixed, 47mL of distilled water is sequentially added into the mixture, the mixture is fully and evenly mixed, different preparations are prepared, and the dispersion condition of the mixture in water is observed. The dispersing agent is sodium tripolyphosphate, sodium lignin sulfonate, sodium dodecyl sulfate, alkyl naphthalene sulfonate and nekal, and the wettability of the bacterial powder after the dispersing agent is added is measured.
TABLE 1 screening results of dispersants in wettable powders
| Name of the name | Dispersibility of | Wetting time/s |
| Alkyl naphthalene sulfonate | +++ | 12.15 |
| Sodium dodecyl sulfate | +++ | 10.79 |
| Sodium lignin sulfonate | +++ | 10.30 |
| Sodium tripolyphosphate | + | 16.31 |
| Pull-open powder | － | 20.12 |
Note that: +++ indicates the most dispersible good (sedimentation rate is less than 10 percent), ++ denotes the secondary dispersibility (sedimentation rate < 20%), + indicates that the dispersibility is general (sedimentation rate < 30%) -indicates that the dispersion is not uniform.
As can be seen from table 1, the wetting time of sodium tripolyphosphate and nekal is long, and the dispersibility is poor; the wetting time of sodium lignin sulfonate, sodium dodecyl sulfate and sodium alkyl naphthalene sulfonate is short, and the dispersing agent has good dispersibility and can be used as a dispersing agent of wettable powder. Thus, sodium alkyl naphthalene sulfonate, sodium dodecyl sulfate, and sodium lignin sulfonate are selected as dispersants for the wettable powder.
Example 3
The screening of the protective agent in the wettable powder for preventing and treating cabbage black spot is described in detail in the embodiment.
The effect of the protective agent is judged by the influence of the protective agent on the bacterial content. Specifically, the protective agent sodium carboxymethyl cellulose, arabic gum, humic acid and skimmed milk powder are added into LB solid medium according to the amount of 5% to prepare LB flat plate, and the flat plate without the protective agent is arranged as a control, and the bacillus megaterium bacterial suspension is diluted to 10 -7 Coating on LB plates containing protective agent and blank control ck1, coating 2 plates for each treatment, turning on ultraviolet lamp to irradiate for 5min after coating, culturing in a constant temperature incubator at 35 ℃ for 24h, counting colony numbers on the plates, setting blank control ck2 plates without ultraviolet irradiation, culturing in a constant temperature incubator at 35 ℃ and counting colony numbers on the plates after 24 h.
The screening results of the protective agent in the wettable powder are shown in table 2. As can be seen from table 2: the colony number of the colony added with the protective agent is better than that of the colony without the protective agent (ck 1) through ultraviolet irradiation treatment; sodium carboxymethyl cellulose, acacia and skimmed milk powder are all preferred over humic acid as the protective agent, but in view of cost, skimmed milk powder is not suitable for mass production and application, so sodium carboxymethyl cellulose and acacia are more preferred to be added as the protective agent of wettable powder.
TABLE 2 screening results of protective agents in wettable powders
| Treatment of | Colony count (number/plate) |
| Sodium carboxymethyl cellulose | 37 |
| Acacia gum | 37.5 |
| Humic acid | 31 |
| Skimmed milk powder | 39 |
| ck1 | 20 |
| ck2 | 52 |
Note that: the colony count in the above table was 2 plates statistically averaged.
Example 4
The preparation process of the wettable powder for preventing and treating cabbage black spot is described in detail in the embodiment.
1 to 10 parts of dispersing agent sodium dodecyl sulfate, 10 to 25 parts of alkyl naphthalene sulfonate, 1 to 6 parts of surfactant sodium lignin sulfonate, 1 to 6 parts of protecting agent Arabic gum, 1 to 5 parts of sodium carboxymethyl cellulose or a combination of two of the above materials and 10 to 25 parts of carrier diatomite are uniformly mixed for standby. Crushing the uniformly mixed auxiliary agent and carrier by an airflow crusher, adding 28-48 parts of bacillus megatherium bacterial powder which is the main component of the crushed auxiliary agent and carrier, and uniformly mixing the crushed auxiliary agent and carrier again. The prepared bacillus megatherium wettable powder after being crushed and uniformly mixed can pass through a mesh screen with 325 meshes, the wetting time is less than 2min, the dispersibility is excellent, and the suspension rate is more than 85%. The spray coating on cabbage leaves after being diluted by 1000 times has better spreadability. Therefore, the preparation method and the formula of the wettable powder for preventing and treating cabbage black spot can be determined according to the proportion, and the formula is shown in the following table 3.
Table 3 formulation of wettable powder for preventing and treating black spot of chinese cabbage
Example 5
The embodiment describes in detail the efficacy experiment of the wettable powder for preventing and treating cabbage black spot.
The test was carried out in a greenhouse from the institute of microbiological fermentation engineering, inc., yunnan province, anning, kunming, inc. A total of 4 treatments were set up, respectively:
A. diluting the wettable powder for preventing and treating cabbage black spot by 1000 times;
B. diluting the wettable powder for preventing and treating cabbage black spot by 2000 times;
C. diluting 70% thiophanate-methyl wettable powder 800 times for treatment;
D. and (5) treating with clear water.
Beginning treatment after the occurrence of Chinese cabbage plants, carrying out treatment for the 2 nd time at intervals of 3 days after spraying for the 1 st time, investigating the disease condition of black spot disease before treatment, carrying out disease investigation 1, 4 and 7 days after treatment, investigating the disease spot areas on all the damaged leaves of the plants, grading according to the disease area, wherein the grading standard is as follows:
level 0: no disease spots;
stage 1: the area of the disease spots accounts for less than 5% of the area of the blade;
3 stages: the area of the disease spots accounts for 6-10% of the area of the leaf blades;
5 stages: the area of the disease spots accounts for 11% -20% of the area of the leaf blades;
7 stages: the area of the disease spots accounts for 21% -40% of the area of the blade;
stage 9: the area of the disease spots accounts for more than 41 percent of the area of the blade;
and calculating the disease index and the control effect according to the disease severity before and after the investigation treatment. The formula is as follows:
as can be seen from table 4: A. b, C has obvious control effect on cabbage black spot, and the control effect of the three treatments is gradually enhanced, and the disease index of the A, B, C treatments is lower than that of the starting treatment except the control group treatment. Wherein after 1d of treatment, the control effect of the treatment C reaches 70.64%, and the quick-acting property is strong; the control effect of the treatment A is inferior and reaches 57.58 percent; the control effect of treatment B was the lowest, 43.14%. After 3d treatment, the statistical results on day 4 are: the prevention effect of the treatment C and the prevention effect of the treatment A are not obviously different and are 78.41 percent and 73.06 percent respectively; the control effect of treatment B was 41.67%, which was different from treatments C and A. And after the treatment is performed for 7 days, the effect of the treatment C is best, the treatment is performed for A times, and the treatment is performed for B times, so that the control effect reaches more than 81%, and the expected control effect is achieved. Thus, it can be derived that: the wettable powder for preventing and treating cabbage black spot has obvious preventing and treating effect on white cabbage black spot, the dilution ratio is preferably 1000-2000, if the land with serious disease is diluted 1000 times, the land with light disease is diluted 2000 times, and the occurrence and diffusion of cabbage black spot can be prevented and treated.
TABLE 4 statistical Table of control effects of different treatments on black spot of Chinese cabbage
Note that: the different lower case letters after the same column of data in the table indicate a significant difference in 0.05 level.
The foregoing is merely illustrative of the present invention, and the present invention is not limited thereto, and any person skilled in the art will readily recognize that variations or substitutions are within the scope of the present invention. Therefore, the protection scope of the present invention shall be subject to the protection scope of the claims.
Claims (4)
1. The wettable powder for preventing and treating cabbage black spot is characterized by comprising the following components in parts by weight: 25 to 65 parts of bacillus megaterium CRP-4 bacterial powder, 15 to 25 parts of dispersing agent, 5 to 15 parts of surfactant, 1 to 6 parts of protective agent and 10 to 80 parts of carrier,
the bacillus megaterium CRP-4 bacterial powder is prepared by taking bacillus megaterium CRP-4 as a bacterial strain, and performing activation, culture, fermentation, centrifugation and drying,
the effective viable count in the bacillus megaterium CRP-4 bacterial powder is not lower than 1000 hundred million/g,
the dispersing agent is one or two of sodium dodecyl sulfate and alkyl naphthalene sulfonate; the surfactant is sodium lignin sulfonate; the protective agent is one or two of acacia and sodium carboxymethylcellulose, and the carrier is diatomite;
bacillus megaterium (Bacillus megaterium), which is numbered CRP-4, accession number of CGMCC No.18458, and date of preservation of 2019, 09, 02, accession number: china general microbiological culture Collection center, preservation address: no.1 and No. 3 of the north cinquefoil of the morning sun area of beijing city.
2. The method for preparing the wettable powder for preventing and treating cabbage black spot according to claim 1, which is characterized by comprising the following steps:
preparing bacillus megatherium CRP-4 bacterial powder;
25 to 65 parts of bacillus megaterium CRP-4 bacterial powder, 15 to 25 parts of dispersing agent, 5 to 15 parts of surfactant, 1 to 6 parts of protective agent and 10 to 80 parts of carrier are compounded to prepare the wettable powder for preventing and treating the black spot of the celery cabbage.
3. The method for preparing wettable powder for preventing and treating cabbage black spot according to claim 2, wherein the preparation of bacillus megatherium CRP-4 bacterial powder comprises the following steps:
activating strains: activating the bacillus megatherium CRP-4 stored at the temperature of minus 80 ℃ on an LB solid medium for 12 to 16 hours to obtain activated bacillus megatherium CRP-4;
seed liquid preparation: scraping a loop of activated bacillus megatherium CRP-4 by using a sterile inoculating loop, inoculating the bacillus megatherium CRP-4 into 50ml of LB liquid medium, and culturing for 24 hours at the temperature of 35 ℃ and the rotating speed of 160rpm to obtain bacillus megatherium CRP-4 seed liquid;
preparing fermentation liquid: inoculating the bacillus megaterium CRP-4 seed liquid into a workshop fermentation material culture medium according to the volume ratio of the strain to the culture liquid of 1:120, and culturing for 36-48 hours at the temperature of 36.5-37.5 ℃ and the rotating speed of 120rpm/min to obtain bacillus megaterium CRP-4 fermentation liquid, wherein the effective viable count of the bacillus megaterium CRP-4 fermentation liquid is not less than 4.0x10 9 cfu/g；
Centrifuging and drying: pouring the fermentation liquor of the bacillus megatherium CRP-4 into a centrifugal machine through equipment, preparing centrifugal thalli of the bacillus megatherium CRP-4 under the condition that the rotating speed of the centrifugal machine is 4000rpm/min,
adding soluble starch and diatomite into the centrifugal thalli of the bacillus megaterium CRP-4, and transferring the centrifugal thalli of the bacillus megaterium CRP-4 added with the soluble starch and the diatomite into spray drying equipment to obtain bacillus megaterium CRP-4 bacterial powder.
4. The application of the wettable powder for preventing and treating cabbage black spot according to claim 1, wherein the wettable powder for preventing and treating cabbage black spot is diluted 1000-2000 times before or at the beginning of the disease, and sprayed on leaf surfaces.
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