CN106396863A - Composite microbial fertilizer for control of botrytis cinerea and preparation method thereof - Google Patents
Composite microbial fertilizer for control of botrytis cinerea and preparation method thereof Download PDFInfo
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- CN106396863A CN106396863A CN201610746812.XA CN201610746812A CN106396863A CN 106396863 A CN106396863 A CN 106396863A CN 201610746812 A CN201610746812 A CN 201610746812A CN 106396863 A CN106396863 A CN 106396863A
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- 239000003337 fertilizer Substances 0.000 title claims abstract description 41
- 238000002360 preparation method Methods 0.000 title claims abstract description 38
- 239000002131 composite material Substances 0.000 title claims abstract description 36
- 230000000813 microbial effect Effects 0.000 title abstract description 13
- 241000123650 Botrytis cinerea Species 0.000 title abstract description 12
- 238000000855 fermentation Methods 0.000 claims abstract description 120
- 230000004151 fermentation Effects 0.000 claims abstract description 120
- 235000007688 Lycopersicon esculentum Nutrition 0.000 claims abstract description 45
- 240000003768 Solanum lycopersicum Species 0.000 claims abstract description 45
- 241000193830 Bacillus <bacterium> Species 0.000 claims abstract description 35
- 241000223259 Trichoderma Species 0.000 claims abstract description 31
- 239000002994 raw material Substances 0.000 claims abstract description 4
- 239000007788 liquid Substances 0.000 claims description 64
- 239000002609 medium Substances 0.000 claims description 54
- 241001655322 Streptomycetales Species 0.000 claims description 49
- 239000007787 solid Substances 0.000 claims description 45
- CSNNHWWHGAXBCP-UHFFFAOYSA-L Magnesium sulfate Chemical compound [Mg+2].[O-][S+2]([O-])([O-])[O-] CSNNHWWHGAXBCP-UHFFFAOYSA-L 0.000 claims description 40
- 238000009423 ventilation Methods 0.000 claims description 38
- 241000894006 Bacteria Species 0.000 claims description 37
- 230000001580 bacterial effect Effects 0.000 claims description 37
- 239000000843 powder Substances 0.000 claims description 30
- 102000002322 Egg Proteins Human genes 0.000 claims description 29
- 108010000912 Egg Proteins Proteins 0.000 claims description 29
- 210000004681 ovum Anatomy 0.000 claims description 29
- 239000001963 growth medium Substances 0.000 claims description 26
- 230000001954 sterilising effect Effects 0.000 claims description 24
- 235000020289 caffè mocha Nutrition 0.000 claims description 23
- 229910052943 magnesium sulfate Inorganic materials 0.000 claims description 20
- 235000019341 magnesium sulphate Nutrition 0.000 claims description 20
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Chemical compound O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 19
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 claims description 16
- 238000000034 method Methods 0.000 claims description 15
- 238000004321 preservation Methods 0.000 claims description 15
- 238000004659 sterilization and disinfection Methods 0.000 claims description 15
- 238000003756 stirring Methods 0.000 claims description 15
- 229920001817 Agar Polymers 0.000 claims description 14
- 239000008272 agar Substances 0.000 claims description 14
- 241000193755 Bacillus cereus Species 0.000 claims description 12
- VTYYLEPIZMXCLO-UHFFFAOYSA-L Calcium carbonate Chemical compound [Ca+2].[O-]C([O-])=O VTYYLEPIZMXCLO-UHFFFAOYSA-L 0.000 claims description 12
- 235000012054 meals Nutrition 0.000 claims description 12
- ZPWVASYFFYYZEW-UHFFFAOYSA-L dipotassium hydrogen phosphate Chemical compound [K+].[K+].OP([O-])([O-])=O ZPWVASYFFYYZEW-UHFFFAOYSA-L 0.000 claims description 11
- 235000015097 nutrients Nutrition 0.000 claims description 11
- FGIUAXJPYTZDNR-UHFFFAOYSA-N potassium nitrate Chemical compound [K+].[O-][N+]([O-])=O FGIUAXJPYTZDNR-UHFFFAOYSA-N 0.000 claims description 10
- WQZGKKKJIJFFOK-GASJEMHNSA-N Glucose Natural products OC[C@H]1OC(O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-GASJEMHNSA-N 0.000 claims description 9
- 239000008103 glucose Substances 0.000 claims description 9
- 238000011081 inoculation Methods 0.000 claims description 9
- 239000002054 inoculum Substances 0.000 claims description 9
- 229940099596 manganese sulfate Drugs 0.000 claims description 9
- 239000011702 manganese sulphate Substances 0.000 claims description 9
- 235000007079 manganese sulphate Nutrition 0.000 claims description 9
- SQQMAOCOWKFBNP-UHFFFAOYSA-L manganese(II) sulfate Chemical compound [Mn+2].[O-]S([O-])(=O)=O SQQMAOCOWKFBNP-UHFFFAOYSA-L 0.000 claims description 9
- 235000013379 molasses Nutrition 0.000 claims description 9
- 239000002504 physiological saline solution Substances 0.000 claims description 9
- 239000004575 stone Substances 0.000 claims description 9
- FRXSZNDVFUDTIR-UHFFFAOYSA-N 6-methoxy-1,2,3,4-tetrahydroquinoline Chemical compound N1CCCC2=CC(OC)=CC=C21 FRXSZNDVFUDTIR-UHFFFAOYSA-N 0.000 claims description 8
- 229920002472 Starch Polymers 0.000 claims description 8
- 239000011780 sodium chloride Substances 0.000 claims description 8
- 239000008107 starch Substances 0.000 claims description 8
- 235000019698 starch Nutrition 0.000 claims description 8
- 241000187747 Streptomyces Species 0.000 claims description 7
- 240000002791 Brassica napus Species 0.000 claims description 6
- 235000004977 Brassica sinapistrum Nutrition 0.000 claims description 6
- 229920002261 Corn starch Polymers 0.000 claims description 6
- 239000001888 Peptone Substances 0.000 claims description 6
- 108010080698 Peptones Proteins 0.000 claims description 6
- 230000033228 biological regulation Effects 0.000 claims description 6
- 229910000019 calcium carbonate Inorganic materials 0.000 claims description 6
- 239000008120 corn starch Substances 0.000 claims description 6
- 235000012343 cottonseed oil Nutrition 0.000 claims description 6
- 229910000402 monopotassium phosphate Inorganic materials 0.000 claims description 6
- 235000019796 monopotassium phosphate Nutrition 0.000 claims description 6
- 235000019319 peptone Nutrition 0.000 claims description 6
- PJNZPQUBCPKICU-UHFFFAOYSA-N phosphoric acid;potassium Chemical compound [K].OP(O)(O)=O PJNZPQUBCPKICU-UHFFFAOYSA-N 0.000 claims description 6
- 239000012153 distilled water Substances 0.000 claims description 5
- 239000011790 ferrous sulphate Substances 0.000 claims description 5
- 235000003891 ferrous sulphate Nutrition 0.000 claims description 5
- BAUYGSIQEAFULO-UHFFFAOYSA-L iron(2+) sulfate (anhydrous) Chemical compound [Fe+2].[O-]S([O-])(=O)=O BAUYGSIQEAFULO-UHFFFAOYSA-L 0.000 claims description 5
- 229910000359 iron(II) sulfate Inorganic materials 0.000 claims description 5
- 235000010333 potassium nitrate Nutrition 0.000 claims description 5
- 239000004323 potassium nitrate Substances 0.000 claims description 5
- OAICVXFJPJFONN-UHFFFAOYSA-N Phosphorus Chemical compound [P] OAICVXFJPJFONN-UHFFFAOYSA-N 0.000 claims description 4
- 239000011574 phosphorus Substances 0.000 claims description 4
- 229910052698 phosphorus Inorganic materials 0.000 claims description 4
- 239000007921 spray Substances 0.000 claims description 4
- 235000017060 Arachis glabrata Nutrition 0.000 claims description 3
- 244000105624 Arachis hypogaea Species 0.000 claims description 3
- 235000010777 Arachis hypogaea Nutrition 0.000 claims description 3
- 235000018262 Arachis monticola Nutrition 0.000 claims description 3
- 244000017020 Ipomoea batatas Species 0.000 claims description 3
- 235000002678 Ipomoea batatas Nutrition 0.000 claims description 3
- PWHULOQIROXLJO-UHFFFAOYSA-N Manganese Chemical compound [Mn] PWHULOQIROXLJO-UHFFFAOYSA-N 0.000 claims description 3
- 240000003183 Manihot esculenta Species 0.000 claims description 3
- 235000016735 Manihot esculenta subsp esculenta Nutrition 0.000 claims description 3
- 244000046052 Phaseolus vulgaris Species 0.000 claims description 3
- 235000010627 Phaseolus vulgaris Nutrition 0.000 claims description 3
- 244000000231 Sesamum indicum Species 0.000 claims description 3
- 235000003434 Sesamum indicum Nutrition 0.000 claims description 3
- 229930006000 Sucrose Natural products 0.000 claims description 3
- CZMRCDWAGMRECN-UGDNZRGBSA-N Sucrose Chemical compound O[C@H]1[C@H](O)[C@@H](CO)O[C@@]1(CO)O[C@@H]1[C@H](O)[C@@H](O)[C@H](O)[C@@H](CO)O1 CZMRCDWAGMRECN-UGDNZRGBSA-N 0.000 claims description 3
- NINIDFKCEFEMDL-UHFFFAOYSA-N Sulfur Chemical compound [S] NINIDFKCEFEMDL-UHFFFAOYSA-N 0.000 claims description 3
- 229940041514 candida albicans extract Drugs 0.000 claims description 3
- 239000006071 cream Substances 0.000 claims description 3
- KCIDZIIHRGYJAE-YGFYJFDDSA-L dipotassium;[(2r,3r,4s,5r,6r)-3,4,5-trihydroxy-6-(hydroxymethyl)oxan-2-yl] phosphate Chemical compound [K+].[K+].OC[C@H]1O[C@H](OP([O-])([O-])=O)[C@H](O)[C@@H](O)[C@H]1O KCIDZIIHRGYJAE-YGFYJFDDSA-L 0.000 claims description 3
- 229910052739 hydrogen Inorganic materials 0.000 claims description 3
- 239000001257 hydrogen Substances 0.000 claims description 3
- 125000004435 hydrogen atom Chemical class [H]* 0.000 claims description 3
- 229910052748 manganese Inorganic materials 0.000 claims description 3
- 239000011572 manganese Substances 0.000 claims description 3
- 239000000203 mixture Substances 0.000 claims description 3
- 235000020232 peanut Nutrition 0.000 claims description 3
- 229920001592 potato starch Polymers 0.000 claims description 3
- 239000000243 solution Substances 0.000 claims description 3
- 239000005720 sucrose Substances 0.000 claims description 3
- 239000011593 sulfur Substances 0.000 claims description 3
- 229910052717 sulfur Inorganic materials 0.000 claims description 3
- 239000012138 yeast extract Substances 0.000 claims description 3
- 150000001875 compounds Chemical class 0.000 claims 1
- 230000000694 effects Effects 0.000 abstract description 20
- 244000005700 microbiome Species 0.000 abstract description 7
- 239000003242 anti bacterial agent Substances 0.000 abstract description 6
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- 229930000044 secondary metabolite Natural products 0.000 abstract description 5
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- 206010059866 Drug resistance Diseases 0.000 abstract description 3
- 239000004098 Tetracycline Substances 0.000 abstract description 3
- 229960002180 tetracycline Drugs 0.000 abstract description 3
- 235000019364 tetracycline Nutrition 0.000 abstract description 3
- 229930101283 tetracycline Natural products 0.000 abstract description 3
- 150000003522 tetracyclines Chemical class 0.000 abstract description 3
- 229960004475 chlortetracycline Drugs 0.000 abstract description 2
- ZGEGCLOFRBLKSE-UHFFFAOYSA-N 1-Heptene Chemical compound CCCCCC=C ZGEGCLOFRBLKSE-UHFFFAOYSA-N 0.000 abstract 1
- 241001249117 Bacillus mojavensis Species 0.000 abstract 1
- 241000970298 Streptomyces champavatii Species 0.000 abstract 1
- 244000052616 bacterial pathogen Species 0.000 abstract 1
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- 244000144972 livestock Species 0.000 abstract 1
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- 210000003608 fece Anatomy 0.000 description 3
- 239000010871 livestock manure Substances 0.000 description 3
- 239000000126 substance Substances 0.000 description 3
- 241000186361 Actinobacteria <class> Species 0.000 description 2
- 241000233866 Fungi Species 0.000 description 2
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- CYDMQBQPVICBEU-UHFFFAOYSA-N chlorotetracycline Natural products C1=CC(Cl)=C2C(O)(C)C3CC4C(N(C)C)C(O)=C(C(N)=O)C(=O)C4(O)C(O)=C3C(=O)C2=C1O CYDMQBQPVICBEU-UHFFFAOYSA-N 0.000 description 2
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- DHPRQBPJLMKORJ-XRNKAMNCSA-N (4s,4as,5as,6s,12ar)-7-chloro-4-(dimethylamino)-1,6,10,11,12a-pentahydroxy-6-methyl-3,12-dioxo-4,4a,5,5a-tetrahydrotetracene-2-carboxamide Chemical compound C1=CC(Cl)=C2[C@](O)(C)[C@H]3C[C@H]4[C@H](N(C)C)C(=O)C(C(N)=O)=C(O)[C@@]4(O)C(=O)C3=C(O)C2=C1O DHPRQBPJLMKORJ-XRNKAMNCSA-N 0.000 description 1
- DHPRQBPJLMKORJ-RQWFMOQPSA-N (4s,4as,6s,12ar)-7-chloro-4-(dimethylamino)-1,6,10,11,12a-pentahydroxy-6-methyl-3,12-dioxo-4,4a,5,5a-tetrahydrotetracene-2-carboxamide Chemical compound C1=CC(Cl)=C2[C@](O)(C)C3C[C@H]4[C@H](N(C)C)C(=O)C(C(N)=O)=C(O)[C@@]4(O)C(=O)C3=C(O)C2=C1O DHPRQBPJLMKORJ-RQWFMOQPSA-N 0.000 description 1
- YZEUHQHUFTYLPH-UHFFFAOYSA-N 2-nitroimidazole Chemical compound [O-][N+](=O)C1=NC=CN1 YZEUHQHUFTYLPH-UHFFFAOYSA-N 0.000 description 1
- 241000589220 Acetobacter Species 0.000 description 1
- 241000251468 Actinopterygii Species 0.000 description 1
- 241001465180 Botrytis Species 0.000 description 1
- 241000222120 Candida <Saccharomycetales> Species 0.000 description 1
- 241001530515 Candida sake Species 0.000 description 1
- 239000004099 Chlortetracycline Substances 0.000 description 1
- 239000005784 Fluoxastrobin Substances 0.000 description 1
- 241000238631 Hexapoda Species 0.000 description 1
- 241001465754 Metazoa Species 0.000 description 1
- RFCWHQNNCOJYTR-IRCAEPKSSA-N Tautomycin Chemical compound O([C@H]([C@H](CC1)C)[C@H](C)CC[C@H](O)[C@H](C)C(=O)C[C@@H](O)[C@@H](OC)[C@H](OC(=O)C[C@@H](O)C=2C(OC(=O)C=2C)=O)C(C)C)[C@@]21CC[C@@H](C)[C@H](CC[C@H](C)C(C)=O)O2 RFCWHQNNCOJYTR-IRCAEPKSSA-N 0.000 description 1
- RFCWHQNNCOJYTR-UHFFFAOYSA-N Tautomycin Natural products CC=1C(=O)OC(=O)C=1C(O)CC(=O)OC(C(C)C)C(OC)C(O)CC(=O)C(C)C(O)CCC(C)C(C(CC1)C)OC21CCC(C)C(CCC(C)C(C)=O)O2 RFCWHQNNCOJYTR-UHFFFAOYSA-N 0.000 description 1
- 241000223261 Trichoderma viride Species 0.000 description 1
- 235000009392 Vitis Nutrition 0.000 description 1
- 241000219095 Vitis Species 0.000 description 1
- 230000003281 allosteric effect Effects 0.000 description 1
- 241000702539 bacterium W10 Species 0.000 description 1
- 230000008827 biological function Effects 0.000 description 1
- 238000009395 breeding Methods 0.000 description 1
- 230000001488 breeding effect Effects 0.000 description 1
- 230000008859 change Effects 0.000 description 1
- 239000003795 chemical substances by application Substances 0.000 description 1
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- 238000004040 coloring Methods 0.000 description 1
- 230000000052 comparative effect Effects 0.000 description 1
- 239000012895 dilution Substances 0.000 description 1
- 238000010790 dilution Methods 0.000 description 1
- 230000007613 environmental effect Effects 0.000 description 1
- 238000003912 environmental pollution Methods 0.000 description 1
- 230000003628 erosive effect Effects 0.000 description 1
- 239000000284 extract Substances 0.000 description 1
- UFEODZBUAFNAEU-NLRVBDNBSA-N fluoxastrobin Chemical compound C=1C=CC=C(OC=2C(=C(OC=3C(=CC=CC=3)Cl)N=CN=2)F)C=1C(=N/OC)\C1=NOCCO1 UFEODZBUAFNAEU-NLRVBDNBSA-N 0.000 description 1
- 238000003306 harvesting Methods 0.000 description 1
- 230000006698 induction Effects 0.000 description 1
- 230000003993 interaction Effects 0.000 description 1
- AXONCVPTTFHNOI-UHFFFAOYSA-N kairine Chemical compound C1=CC(O)=C2N(C)CCCC2=C1 AXONCVPTTFHNOI-UHFFFAOYSA-N 0.000 description 1
- 230000002045 lasting effect Effects 0.000 description 1
- 206010025482 malaise Diseases 0.000 description 1
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- 239000000375 suspending agent Substances 0.000 description 1
Classifications
-
- C—CHEMISTRY; METALLURGY
- C05—FERTILISERS; MANUFACTURE THEREOF
- C05G—MIXTURES OF FERTILISERS COVERED INDIVIDUALLY BY DIFFERENT SUBCLASSES OF CLASS C05; MIXTURES OF ONE OR MORE FERTILISERS WITH MATERIALS NOT HAVING A SPECIFIC FERTILISING ACTIVITY, e.g. PESTICIDES, SOIL-CONDITIONERS, WETTING AGENTS; FERTILISERS CHARACTERISED BY THEIR FORM
- C05G3/00—Mixtures of one or more fertilisers with additives not having a specially fertilising activity
-
- C—CHEMISTRY; METALLURGY
- C05—FERTILISERS; MANUFACTURE THEREOF
- C05F—ORGANIC FERTILISERS NOT COVERED BY SUBCLASSES C05B, C05C, e.g. FERTILISERS FROM WASTE OR REFUSE
- C05F11/00—Other organic fertilisers
- C05F11/08—Organic fertilisers containing added bacterial cultures, mycelia or the like
-
- 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
- Y02A40/00—Adaptation technologies in agriculture, forestry, livestock or agroalimentary production
- Y02A40/10—Adaptation technologies in agriculture, forestry, livestock or agroalimentary production in agriculture
- Y02A40/22—Improving land use; Improving water use or availability; Controlling erosion
Landscapes
- Chemical & Material Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Organic Chemistry (AREA)
- Pest Control & Pesticides (AREA)
- Biochemistry (AREA)
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
- Agricultural Chemicals And Associated Chemicals (AREA)
- Fertilizers (AREA)
Abstract
The invention discloses a composite microbial fertilizer for control of botrytis cinerea and a preparation method thereof. The composite microbial fertilizer is prepared by mixing of the following raw materials: a Str. Viridifaciens fermentation broth, a S. champavatii fermentation broth, an India bacillus fermentation broth, a B. mojavensis fermentation broth, and a Trichoderma ovalisporium fermentation broth. The strains involved in the invention can comprehensively utilize the antibiosis, competition, hyperparasitism and bacteriolysis effects between or within microbial species, or utilize secondary metabolites to induce the disease resistance of tomatoes, and can generate a variety of antibiotics like tetracycline, 7-aureomycin, champamycin A and B (antifungal antibiotics of heptaene) and champavatin to inhibit botrytis cinerea pathogenic bacteria and strengthen its disease prevention effect. The composite microbial fertilizer provided by the invention is safe to human and livestock, is environment-friendly, and does not produce drug resistance when applied to control of botrytis cinerea. The composite microbial fertilizer has the advantages of simple preparation method, low cost, and easy use.
Description
Technical field
The invention belongs to Strategies of Agricultural Bio-control and microbial manure technical field are and in particular to a kind of prevent and treat graw mold of tomato
Composite microbic bacterial fertilizer and preparation method thereof.
Background technology
Graw mold of tomato, as the fungal disease of the tomato production that causes harm in modern cultivation, often results in huge loss.I
State localised protection ground Fructus Lycopersici esculenti is subject to the gray mold serious plot underproduction of being caused harm to be up to more than 60%；Many tomato in greenhouse are subject to gray mold
Cause harm and lead to underproduction 20%-30%, serious plot is more than 50%；Shandong Province Jiaodong Areas Fructus Lycopersici esculenti young fruit is encroached on by ash arrhizus bacteria
Underproduction 10%-30%, serious plot even has no harvest.
Tomato gray mould be had a strong impact on by a kind of microbial global fungal disease of Botrytis cinerea Fructus Lycopersici esculenti yield and
Quality.At present, the main prevention and controls of graw mold of tomato are had:One is chemical prevention.Though chemical prevention has instant effect, efficiency high
The advantages of, but because life-time service can cause the shortcomings of environmental pollution, natural enemy of harm insect, therefore progressively it is eliminated;Secondly
Biological control, due to Biological control drug effect height, lasting period length, be not likely to produce Drug resistance, advantages of environment protection is increasingly subject to
The favor of people.
The Biological control of plant disease is to adjust the microbial environment of host crop in Agro-ecological System so as to be beneficial to
Host crop and be unfavorable for pathogen or make it be conducive to host to be unfavorable for the interaction of host and pathogen
The impact of pathogen, thus reach the purpose of controlling disease.The essence of Biological control be using microbial species between or anti-in planting
Life, competition, superparasitism, bacteriolysiss, or induce plant to produce disease resistance etc. by microbial secondary metabolite, to suppress
The survival of some pathogens and activity.It is a kind of friendly to environmental ecology, continuable Field Plants pest control method, more
More to obtain various countries and to plant recognizing and paying attention to of sick worker, oneself obtains significant progress.In recent years, Biological control is in Fructus Lycopersici esculenti ash
Mould prevent and treat aspect, constantly deeply and expand.Be presently used for prevent and treat graw mold of tomato microorganism mainly include funguses,
Antibacterial and actinomycetes three major types.Prevent and treat nearly 10 kinds of the biocontrol fungi of gray mold, study and apply wider mainly Trichoderma spp. and ferment
Female bacterium.Pan Yani etc. reaches 92.6% with Trichoderma viride preventing and treating greenhouse tomato gray mold prevention effect, compared with untreated plant,
Sickness rate reduces 39.1%；Wilson etc. makes grey Fructus Vitis viniferae embrace the Fructus Mali pumilae causing using very thin candida mycoderma and candida sake
Wound rots to reduce 50%.The antibacterial of gray mold study on prevention and application also has kind more than 10, and bacillus cereuss are then current grey mold
The main bacteria bacterial strain studied in bacterium Biological control and apply.The Antagonism that Tong Yunhui etc. separates acquisition is strong, the ground of has a broad antifungal spectrum
Clothing bud pole bacterium W10, W3, Y2-11-1 bacterial strain, the preventive effect to graw mold of tomato blade and fruit is 70%-80%, better than 50% speed
2000 times of liquid of kairine.Actinomycetes be used for prevent and treat tomato gray mould mainly use Streptomycetaceae streptomyces generation Wuyiencin,
The antibiotic such as mycin when phosphorus azomycin, Bai Yuetai mycin, Tautomycin, allosteric rhzomorph and fish, all has preferable suppression to gray mold
Make and use.The bacillus cereuss being presently used for preventing and treating graw mold of tomato mainly have the CGMCC No.4777 (patent No.:
ZL201210059785.0), BA-KA3 (number of patent application:201410618581. 5), SZ23 (number of patent application:
201410683826. 2), LH-1 (number of patent application:201410735508.6) and NCPSJI7 (number of patent application:
201310112580.9) etc..Due to multiformity and the synchronous evolution characteristic of pathogen, single antagonistic strain is also unable to reach to be managed very much
The prevention effect thought is combined forming the group that effect preferably prevents and treats graw mold of tomato it is therefore desirable to find different types of bacterium
Close.Due to multiformity and the synchronous evolution characteristic of pathogen, single antagonistic strain is also unable to reach highly desirable prevention effect, because
This, a lot of biological functions of biocontrol microorganisms need to rely on the synergism between two plants or more than two plants of antibacterial, and competence exertion goes out more
Good effect.
Content of the invention
The first object of the present invention is to provide a kind of composite microbic bacterial fertilizer of preventing and treating graw mold of tomato.This composite microbial
The proportioning raw materials of thing bacterial manure are：Raw green streptomycete fermentation liquid 10-30 part, prosperous handkerchief watt bottom streptomycete fermentation liquid 20-40 part, India's bud
Spore bacillus fermentation liquid 10-30 part, mocha husband's fermentation of bacillus liquid 10-30 part, ovum spore Trichoderma spp. fermentation liquid 10-30 part.Described
In microbial bacterial agent, raw green streptomycete, prosperous handkerchief watt bottom streptomycete, India's bacillus cereuss, mocha husband's bacillus cereuss, ovum spore Trichoderma spp.
Five kinds of bacterium are the energy symbiotic co-existences that screening obtains from more than 400 plants of microbial strains, can produce multiple antibiotics such as tetracycline, 7-
Chlortetracycline, champamycin A and B（Seven alkene antifungal antibiotics）With prosperous handkerchief watt bottom rhzomorph；They are to suppression graw mold of tomato --
The pathogen of Botrytis cinerea has especially powerful effect, thus reaching the effect of preventing and treating graw mold of tomato；On the other hand, the strain of the present invention
Comprehensively using the antibiosis between microbial species or in kind, competition, superparasitism, bacteriolysiss, or can be produced by cometabolism
Thing induction Fructus Lycopersici esculenti produces disease resistance, strengthens its protection effect；Again, 5 plants of bacterial strains of this used in the present invention are all can be from soil
Be directly isolated to obtain in earth, all there are root mark growth-promoting functions, can crop surface, plant be internal or soil in flourish same
When, and orienting the plant rhizosphere some secondary metabolites of secretion, it is possible to increase plant rises to the absorption of nutrient, stimulation plant strain growth
Effect to fertilizer.
The second object of the present invention is to provide a kind of preparation method of the composite microbic bacterial fertilizer of preventing and treating graw mold of tomato,
The method step is simple it is ensured that the medicine fertilizer efficiency fruit of microbial-bacterial fertilizer is prominent.
In order to achieve the above object, the preparation method of the composite microbic bacterial fertilizer of this preventing and treating graw mold of tomato, including following
Step：
Step one：The preparation of raw green streptomycete fermentation liquid
Take out raw green streptomyces species preservation pipe, draw flat board with Gause I solid medium and recovered, cultivate 7 days for 30 DEG C.
Under flat board, the streak inoculation of picking single bacterium colony is equipped with the Fructus Solani melongenae bottle of 150 milliliters of Gause I solid mediums, in incubator
In 30 DEG C cultivate 6-8 days, treat that lawn covers with Fructus Solani melongenae bottle, produce a large amount of spore powders and can use 500 milliliters of physiological saline solution to wash
De-, regulation spore concentration is 0.1 hundred million cfu/ml, that is, green streptomycete seed liquor of making a living；
Fermentation：Green for the life of above-mentioned preparation streptomycete seed liquor is seeded to the green chain of life of the 600L equipped with sterilizing with 1% inoculum concentration
In the 1000L fermentation tank of mold fermentation culture medium, 28-32 DEG C of temperature control, first 24 hours, ventilation be per minute for 200L air,
24-36 hour, ventilation is 400L, and after 36 hours, ventilation is 600L, opens stirring 200r/min, cultivates 40-48 hour, treat
Thalline content reaches 60g/L, you can stop tank, that is, obtain raw green streptomycete fermentation liquid；
Wherein, described Gause I solid medium：Potassium nitrate：1 gram, soluble starch：20 grams, dipotassium hydrogen phosphate：0.5
Gram, magnesium sulfate：0.5 gram, sodium chloride：0.5 gram, ferrous sulfate：0.01 gram, agar：20 grams, 1000ml supplied by distilled water,
PH7.2～7.4；
Wherein, described life green streptomycete fermentation culture medium：Peanut meal powder 3%, glucose 1%, stone powder 1%, soy molasses 2%, phosphorus
Sour hydrogen dipotassium 0.04%, magnesium sulfate 0.05%, the condition of each medium sterilization is：0.10-0.15MPa, 121 DEG C sterilize 30 minutes；
Step 2：The preparation of prosperous handkerchief watt bottom streptomycete fermentation liquid
Take out prosperous handkerchief watt bottom streptomyces species preservation pipe, draw flat board with Gause I solid medium and recovered, 30 DEG C of cultures 7
My god.Under flat board, the streak inoculation of picking single bacterium colony, equipped with the Fructus Solani melongenae bottle of 150 milliliters of Gause I solid mediums, is being cultivated
Cultivate 6-8 days for 30 DEG C in case, treat that lawn covers with Fructus Solani melongenae bottle, produce the physiological saline solution that a large amount of spore powders can use 500 milliliters
Eluting, regulation spore concentration is 0.1 hundred million cfu/ml, as prosperous handkerchief watt bottom streptomycete seed liquor；
Fermentation：The prosperous handkerchief watt bottom streptomycete seed liquor of above-mentioned preparation is seeded to prosperous equipped with the 600L sterilizing with 1% inoculum concentration
In the 1000L fermentation tank of handkerchief watt bottom streptomycete fermentation culture medium, 28-30 DEG C of temperature control, first 18 hours, ventilation was per minute to be
300L air, 18-32 hour, ventilation is 500L, and after 32 hours, ventilation is 700L, opens stirring 200r/min, cultivates 40-
48 hours, treat that thalline content reaches 80g/L, you can stop tank, that is, obtain prosperous handkerchief watt bottom streptomycete fermentation liquid；
Wherein, described Gause I solid medium：Potassium nitrate：1 gram, soluble starch：20 grams, dipotassium hydrogen phosphate：0.5
Gram, magnesium sulfate：0.5 gram, sodium chloride：0.5 gram, ferrous sulfate：0.01 gram, agar：20 grams, 1000ml supplied by distilled water,
PH7.2～7.4；
Wherein, described prosperous handkerchief watt bottom streptomycete fermentation culture medium：Sesame seed meal powder 3%, cottonseed meal 1%, corn starch 2%, stone powder 1%,
Cane molasses 3%, manganese sulfate 0.002%, dipotassium hydrogen phosphate 0.01%, magnesium sulfate 0.02%, the condition of each medium sterilization is：
0.10-0.15MPa, 121 DEG C sterilize 30 minutes.
Step 3, the preparation of India's fermentation of bacillus liquid
Take out India's bacillus cereuss preservation pipe, draw flat board respectively with LB solid medium and recovered, cultivate 48 hours for 30 DEG C.?
Under flat board, picking single bacterium colony is seeded to equipped with LB solid medium, culture 48 hours in 30 DEG C of incubators, with 3000ml no
Bacterium water, by the lawn eluting in three Fructus Solani melongenae bottles, is seeded to the 500L fermentation tank equipped with 300L India fermentation of bacillus culture medium
In, open stirring 120r/min, ventilation is 200L/min within first 12 hours, after 12 hours, ventilation is 320L/min, 30 DEG C of cultures
26-36 hour, treats that total spore content is not less than 6,000,000,000 cfu/ml, you can as India's fermentation of bacillus liquid；
Wherein, described LB culture medium：Yeast extract 5.0 g, peptone 10.0, NaCl10.0 g, agar 20 g, water
1000mL, pH7.2；
Wherein, described India fermentation of bacillus culture medium：Glucose 8g/L, corn starch 22 g/L, bean cake powder 50 g/L,
Magnesium sulfate 0.4 g/L, manganese sulfate 0.5 g/L, potassium dihydrogen phosphate 0.5 g/L, Calcium Carbonate 5 g/L, pH7.0.Each medium sterilization
Condition be：0.10-0.15MPa, 121 DEG C sterilize 30 minutes.
Step 4, the preparation of mocha husband's fermentation of bacillus liquid
Take out mocha husband's bacillus cereuss preservation pipe, draw flat board respectively with nutrient broth solid medium and recovered, 30 DEG C of cultures
48 hours.Under flat board, picking single bacterium colony is seeded to equipped with nutrient broth solid medium, cultivates 48 in 30 DEG C of incubators
Hour, with 3000ml sterilized water by the lawn eluting in three Fructus Solani melongenae bottles, it is seeded to equipped with 300L mocha husband's fermentation of bacillus
In the 500L fermentation tank of culture medium, open stirring 120r/min, ventilation is 200L/min within first 8 hours, and after 8 hours, ventilation is
300L/min, 30 DEG C of culture 24-36 hours, treat that total spore content is not less than 8,000,000,000 cfu/ml, you can as mocha husband's spore bar
Fermented liquid；
Wherein, described nutrient broth solid medium：Peptone 10.0 g/L, Carnis Bovis seu Bubali cream 3.0 g/L, sodium chloride 5.0 g/L,
Agar 20 g/L, pH 7.2 ± 0.2；
Wherein, described mocha husband's fermentation of bacillus culture medium：Glucose 8g/L, tapioca starch 20 g/L, rapeseed cake powder 30 g/L,
Magnesium sulfate 0.4 g/L, manganese sulfate 0.5 g/L, potassium dihydrogen phosphate 0.2 g/L, Calcium Carbonate 5 g/L, pH7.0.Each medium sterilization
Condition be：0.10-0.15MPa, 121 DEG C sterilize 30 minutes.
Step 5, the preparation of ovum spore Trichoderma spp. fermentation liquid
Take out ovum spore Trichoderma spp. strain preservation pipe, draw flat board with PDA solid medium and recovered, cultivate 7 days for 30 DEG C.Under flat board
Picking single bacterium colony streak inoculation, equipped with the Fructus Solani melongenae bottle of 150 milliliters of PDA solid mediums, cultivates 6-8 for 30 DEG C in incubator
My god, treat that lawn covers with Fructus Solani melongenae bottle, produce the physiological saline solution eluting that a large amount of spore powders can use 500 milliliters, adjust spore dense
Spend for 0.05 hundred million cfu/ml, as ovum spore Trichoderma spp. seed liquor；
Fermentation：The ovum spore Trichoderma spp. seed liquor of above-mentioned preparation is seeded to equipped with 300L ovum spore Trichoderma spp. fermentation culture with 2% inoculum concentration
In the 500L fermentation tank of base, open stirring 200r/min, ventilation is 120L/min within first 8 hours, and after 8-24 hour, ventilation is
240L/min, after 24 hours, ventilation is 360L/min, 30 DEG C of culture 32-48 hours, treats that thalline content reaches 80g/L, that is,
Tank can be stopped, you can as ovum spore Trichoderma spp. fermentation liquid；
Wherein, described PDA solid medium：Rhizoma Solani tuber osi 200g, sucrose 20g, water 1000mL, agar 20g；
Wherein, described ovum spore Trichoderma spp. fermentation medium：Rapeseed cake 2%, cottonseed meal 2%, sweet potato flour 2%, stone powder 1%, soy molasses 3%, sulfur
Sour manganese 0.001%, dipotassium hydrogen phosphate 0.04%, magnesium sulfate 0.02%, the condition of each medium sterilization is：0.10-0.15MPa, 121
DEG C sterilizing 30 minutes.
Step 6, mixed fermentation liquid
The bacterium solution of first five step preparation is pressed raw green streptomycete fermentation liquid 10-30 part, prosperous handkerchief watt bottom streptomycete fermentation liquid 20-40
Part, India's fermentation of bacillus liquid 10-30 part, mocha husband's fermentation of bacillus liquid 10-30 part, ovum spore Trichoderma spp. fermentation liquid 10-30
Part.Mix, that is, obtain preventing and treating the composite microbic bacterial fertilizer of graw mold of tomato.
Wherein, this composite microbic bacterial fertilizer is foliage-spray each 2-5L/ mu in the method for preventing and treating graw mold of tomato, and it is dilute
The multiple released is 100-200 times, continuously uses 2-3 time, every 7-10 days using once.
The present invention has the advantage that and beneficial effect：, the green streptomycete of life that comprises in the present invention, prosperous handkerchief watt bottom streptomycete, print
Degree bacillus cereuss, mocha husband's bacillus cereuss, five kinds of bacterium of ovum spore Trichoderma spp. are to screen the energy obtaining from more than 400 plants of microbial strains
Symbiotic co-existence, can be in the root of plant, stem, in leaf and soil can growth and breeding, multiple antibiotics such as tetracycline, 7- can be produced
Chlortetracycline, champamycin A and B（Seven alkene antifungal antibiotics）With prosperous handkerchief watt bottom rhzomorph；They are to suppression graw mold of tomato --
The pathogen of Botrytis cinerea has especially powerful effect, thus reaching the effect of superpower preventing and treating graw mold of tomato, it is to graw mold of tomato
Drug effect is high, and average preventive effect is 90. more than 0% and with strong points；, the strain of the present invention can comprehensively utilize microbial species
Between or plant in antibiosis, competition, superparasitism, bacteriolysiss, or by secondary metabolite induce Fructus Lycopersici esculenti generation disease resistance, increasing
Its protection effect strong；, this 5 plants of bacterial strains used in the present invention be all can be directly isolated to obtain from soil, all there is root
Mark growth-promoting functions, can be in crop surface, plant inside or soil while flourish, and it is some to orient plant rhizosphere secretion
Secondary metabolite, it is possible to increase plant to the absorption of nutrient, stimulate plant strain growth to play the effect of fertilizer；(4), the present invention
Composite microbic bacterial fertilizer, to people, animal safety, belongs to environmentally friendly, is not likely to produce using the inventive method preventing and treating graw mold of tomato
Drug resistance, preparation method of the present invention is simple, strain is many, and the antagonistic substance of generation is extensive, effect is significant, and low cost, use are simple.
Specific embodiment
Embodiment 1
A kind of composite microbic bacterial fertilizer of preventing and treating graw mold of tomato is it is characterised in that the proportioning raw materials of this microbial-bacterial fertilizer are：Raw
20 parts of green streptomycete fermentation liquid, 30 parts of prosperous handkerchief watt bottom streptomycete fermentation liquid, 20 parts of India's fermentation of bacillus liquid, mocha husband's spore
20 parts of bacillus fermentation liquid, 20 parts of ovum spore Trichoderma spp. fermentation liquid.
The preparation method of the composite microbic bacterial fertilizer of this preventing and treating graw mold of tomato, comprises the following steps：
Step one：The preparation of raw green streptomycete fermentation liquid
Take out raw green streptomyces species preservation pipe, draw flat board with Gause I solid medium and recovered, cultivate 7 days for 30 DEG C.
Under flat board, the streak inoculation of picking single bacterium colony is equipped with the Fructus Solani melongenae bottle of 150 milliliters of Gause I solid mediums, in incubator
In 30 DEG C cultivate 6-8 days, treat that lawn covers with Fructus Solani melongenae bottle, produce a large amount of spore powders and can use 500 milliliters of physiological saline solution to wash
De-, regulation spore concentration is 0.1 hundred million cfu/ml, that is, green streptomycete seed liquor of making a living；
Fermentation：Green for the life of above-mentioned preparation streptomycete seed liquor is seeded to the green chain of life of the 600L equipped with sterilizing with 1% inoculum concentration
In the 1000L fermentation tank of mold fermentation culture medium, 28-32 DEG C of temperature control, first 24 hours, ventilation be per minute for 200L air,
24-36 hour, ventilation is 400L, and after 36 hours, ventilation is 600L, opens stirring 200r/min, cultivates 44 hours, record bacterium
Body burden is 65g/L, stops tank, that is, obtain raw green streptomycete fermentation liquid；
Wherein, described Gause I solid medium：Potassium nitrate：1 gram, soluble starch：20 grams, dipotassium hydrogen phosphate：0.5
Gram, magnesium sulfate：0.5 gram, sodium chloride：0.5 gram, ferrous sulfate：0.01 gram, agar：20 grams, 1000ml supplied by distilled water,
PH7.2～7.4；
Wherein, described life green streptomycete fermentation culture medium：Peanut meal powder 3%, glucose 1%, stone powder 1%, soy molasses 2%, phosphorus
Sour hydrogen dipotassium 0.04%, magnesium sulfate 0.05%, the condition of each medium sterilization is：0.10-0.15MPa, 121 DEG C sterilize 30 minutes.
Step 2：The preparation of prosperous handkerchief watt bottom streptomycete fermentation liquid
Take out prosperous handkerchief watt bottom streptomyces species preservation pipe, draw flat board with Gause I solid medium and recovered, 30 DEG C of cultures 7
My god.Under flat board, the streak inoculation of picking single bacterium colony, equipped with the Fructus Solani melongenae bottle of 150 milliliters of Gause I solid mediums, is being cultivated
Cultivate 6-8 days for 30 DEG C in case, treat that lawn covers with Fructus Solani melongenae bottle, produce the physiological saline solution that a large amount of spore powders can use 500 milliliters
Eluting, regulation spore concentration is 0.1 hundred million cfu/ml, as prosperous handkerchief watt bottom streptomycete seed liquor；
Fermentation：The prosperous handkerchief watt bottom streptomycete seed liquor of above-mentioned preparation is seeded to prosperous equipped with the 600L sterilizing with 1% inoculum concentration
In the 1000L fermentation tank of handkerchief watt bottom streptomycete fermentation culture medium, 28-30 DEG C of temperature control, first 18 hours, ventilation was per minute to be
300L air, 18-32 hour, ventilation is 500L, and after 32 hours, ventilation is 700L, opens stirring 200r/min, and culture 46 is little
When, recording thalline content is 82g/L, stops tank, that is, obtains prosperous handkerchief watt bottom streptomycete fermentation liquid；
Wherein, described Gause I solid medium：Potassium nitrate：1 gram, soluble starch：20 grams, dipotassium hydrogen phosphate：0.5
Gram, magnesium sulfate：0.5 gram, sodium chloride：0.5 gram, ferrous sulfate：0.01 gram, agar：20 grams, 1000ml supplied by distilled water,
PH7.2～7.4；
Wherein, described prosperous handkerchief watt bottom streptomycete fermentation culture medium：Sesame seed meal powder 3%, cottonseed meal 1%, corn starch 2%, stone powder 1%,
Cane molasses 3%, manganese sulfate 0.002%, dipotassium hydrogen phosphate 0.01%, magnesium sulfate 0.02%, the condition of each medium sterilization is：
0.10-0.15MPa, 121 DEG C sterilize 30 minutes.
Step 3, the preparation of India's fermentation of bacillus liquid
Take out India's bacillus cereuss preservation pipe, draw flat board respectively with LB solid medium and recovered, cultivate 48 hours for 30 DEG C.?
Under flat board, picking single bacterium colony is seeded to equipped with LB solid medium, culture 48 hours in 30 DEG C of incubators, with 3000ml no
Bacterium water, by the lawn eluting in three Fructus Solani melongenae bottles, is seeded to the 500L fermentation tank equipped with 300L India fermentation of bacillus culture medium
In, open stirring 120r/min, ventilation is 200L/min within first 12 hours, after 12 hours, ventilation is 320L/min, 30 DEG C of cultures
28 hours, recording total spore content was 6,100,000,000 cfu/ml, that is, as India's fermentation of bacillus liquid；
Wherein, described LB culture medium：Yeast extract 5.0 g, peptone 10.0, NaCl10.0 g, water 1000mL, pH7.2,
Agar 2% is added in solid medium；
Wherein, described India fermentation of bacillus culture medium：Glucose 8g/L, corn starch 22 g/L, bean cake powder 50 g/L,
Magnesium sulfate 0.4 g/L, manganese sulfate 0.5 g/L, potassium dihydrogen phosphate 0.5 g/L, Calcium Carbonate 5 g/L, pH7.0.Each medium sterilization
Condition be：0.10-0.15MPa, 121 DEG C sterilize 30 minutes.
Step 4, the preparation of mocha husband's fermentation of bacillus liquid
Take out mocha husband's bacillus cereuss preservation pipe, draw flat board respectively with nutrient broth solid medium and recovered, 30 DEG C of cultures
48 hours.Under flat board, picking single bacterium colony is seeded to equipped with nutrient broth solid medium, cultivates 48 in 30 DEG C of incubators
Hour, with 3000ml sterilized water by the lawn eluting in three Fructus Solani melongenae bottles, it is seeded to equipped with 300L mocha husband's fermentation of bacillus
In the 500L fermentation tank of culture medium, open stirring 120r/min, ventilation is 200L/min within first 8 hours, and after 8 hours, ventilation is
300L/min, cultivates 29 hours for 30 DEG C, the total spore content of detection is 8,300,000,000 cfu/ml, that is, as mocha husband's fermentation of bacillus
Liquid；
Wherein, described nutrient broth solid medium：Peptone 10.0 g/L, Carnis Bovis seu Bubali cream 3.0 g/L, sodium chloride 5.0 g/L,
Agar 20 g/L, pH 7.2 ± 0.2；
Wherein, described mocha husband's fermentation of bacillus culture medium：Glucose 8g/L, tapioca starch 20 g/L, rapeseed cake powder 30 g/L,
Magnesium sulfate 0.4 g/L, manganese sulfate 0.5 g/L, potassium dihydrogen phosphate 0.2 g/L, Calcium Carbonate 5 g/L, pH7.0.Each medium sterilization
Condition be：0.10-0.15MPa, 121 DEG C sterilize 30 minutes.
Step 5, the preparation of ovum spore Trichoderma spp. fermentation liquid
Take out ovum spore Trichoderma spp. strain preservation pipe, draw flat board with PDA solid medium and recovered, cultivate 7 days for 30 DEG C.Under flat board
Picking single bacterium colony streak inoculation, equipped with the Fructus Solani melongenae bottle of 150 milliliters of PDA solid mediums, cultivates 6-8 for 30 DEG C in incubator
My god, treat that lawn covers with Fructus Solani melongenae bottle, produce the physiological saline solution eluting that a large amount of spore powders can use 500 milliliters, adjust spore dense
Spend for 0.05 hundred million cfu/ml, as ovum spore Trichoderma spp. seed liquor；
Fermentation：The ovum spore Trichoderma spp. seed liquor of above-mentioned preparation is seeded to equipped with 300L ovum spore Trichoderma spp. fermentation culture with 2% inoculum concentration
In the 500L fermentation tank of base, open stirring 200r/min, ventilation is 120L/min within first 8 hours, and after 8-24 hour, ventilation is
240L/min, after 24 hours, ventilation is 360L/min, cultivates 36 hours for 30 DEG C, detection thalline content is 81g/L, stops tank,
Can be used as ovum spore Trichoderma spp. fermentation liquid；
Wherein, described PDA solid medium：Rhizoma Solani tuber osi 200g, sucrose 20g, water 1000mL, agar 20g；
Wherein, described ovum spore Trichoderma spp. fermentation medium：Rapeseed cake 2%, cottonseed meal 2%, sweet potato flour 2%, stone powder 1%, soy molasses 3%, sulfur
Sour manganese 0.001%, dipotassium hydrogen phosphate 0.04%, magnesium sulfate 0.02%, the condition of each medium sterilization is：0.10-0.15MPa, 121
DEG C sterilizing 30 minutes.
Step 6, mixed fermentation liquid
The bacterium solution of first five step preparation is pressed 20 parts of the green streptomycete fermentation liquid of life, 30 parts of prosperous handkerchief watt bottom streptomycete fermentation liquid, print
20 parts of fermentation of bacillus liquid of degree, 20 parts of mocha husband's fermentation of bacillus liquid, 20 parts of ovum spore Trichoderma spp. fermentation liquid.Mix, that is, obtain
The composite microbic bacterial fertilizer of preventing and treating graw mold of tomato.
Embodiment 2
The antagonism test to tomato gray mould bacterium for the composite microbic bacterial fertilizer of the present invention
(1) test period and place:In April, 2014, indoor in the Experiment on Microbiology of Yan Hui bio tech ltd of Foshan City
Carry out；
(2) test method：
(1) for examination tomato gray mould bacterium source：Tomato gray mould bacterium pathogen-the pathogen of Botrytis cinerea is given for agricultural college of Guangxi University, causes
Sick power measures and shows as High pathogenicity；
(2) flat board face-off experiment:
First the pathogen of Botrytis cinerea is activated on PDA plate, in colony edge region after cultivating 4 days, with card punch (6mm) punching
Make bacterium piece, by the switching of the pathogen of Botrytis cinerea bacterium piece in another PDA plate central authorities, then answering the middle preparation of embodiment 1 step (1)
Close microbial inoculum and dilute 100 times, point is connected on away from indicator bacteria bacterium piece 2.5 centimeters, if blank (is not put and connect complex microorganism
The tomato gray mould bacteria growing situation of bacterial manure).25 DEG C of constant temperature culture, when blank will cover with whole culture dish, measurement ash
The comparison increment (colony radius) of botrytis and process increment, antagonism bacteriostasis rate represents.Computing formula is:
Bacteriostasis rate (%)=(comparison increment one processes increment)/comparison increment X 100
Result：As table 1, composite microbic bacterial fertilizer reaches 91.79% to the suppression ratio of the pathogen of Botrytis cinerea；Transparent antibacterial bandwidth reaches 21.
5 millimeters, illustrate that composite microbic bacterial fertilizer of the present invention is strong to tomato gray mould bacterium biocidal property, the biological and ecological methods to prevent plant disease, pests, and erosion with preventing and treating graw mold of tomato is dived
Power.
The antagonism test result to the pathogen of Botrytis cinerea for table 1 composite microbic bacterial fertilizer
| Process | Comparison increment（mm） | Process increment（mm） | Bacteriostasis rate（%） |
| Composite microbic bacterial fertilizer | 39 | 3.2 | 91.79 |
Embodiment 3
Test effect in booth for the composite microbic bacterial fertilizer of preventing and treating graw mold of tomato
(1) test process:
Test group：This composite microbic bacterial fertilizer of embodiment 1 preparation is that foliage-spray is each in the method for preventing and treating graw mold of tomato
4L/ mu, the multiple of its dilution is 100-200 times, continuously uses 2 times, every 7 days using once；
Chemical agent：Fluoxastrobin suspending agent (250 g/l) (Syngenta Co., Ltd of Britain), 500 times of dilute with water；
Blank：Clear water
(2) test method：July March one in 2015 is carried out in Cangzhou, Hebei Province Cang County Fructus Lycopersici esculenti booth.Start whole during test
In individual canopy, Fructus Lycopersici esculenti leaf portion gray mold moderate occurs, and extracts sick leaf before dispenser.Every cell three row, four repetitions, random district's groups arrangement.
Composite microbic bacterial fertilizer prepared by embodiment 1 is inoculated using knapsack electric sprayer；Separately set comparison medicament and blank.
Dispenser first sprays 1 time after 7 days again, the 7th after secondary dispenser day investigate each process cell individual plant disease the number of sheets with pluck into
Ripe fruit, and calculate preventive effect and rate of growth；
Result：From table 2 it can be seen that after composite microbic bacterial fertilizer is processed, the individual plant disease number of sheets of its graw mold of tomato is substantially less than
Blank and chemical agent comparison, its preventive effect reaches 93. 12%, and the composite microbic bacterial fertilizer that the present invention is described is to tomato gray mould
Disease has good prevention effect；
In addition, as can be seen from Table 2, the tomato yield applying the composite microbic bacterial fertilizer of the present invention is significantly larger than blank control group,
Rate of growth has reached 40.45%, also high than its yield by 22.55% with the contrast of chemical agent group, and finds that its fruit is big and full,
Number of results is many, and heterocarp is few, in good taste, and coloring is vivid, and its fruit sweetness is also obviously improved.
The comparative test result to graw mold of tomato preventive effect for table 2 composite microbic bacterial fertilizer
| Process | The individual plant disease number of sheets（Piece） | Preventive effect（%） | Yield（kg） | Rate of growth（%） |
| Composite microbic bacterial fertilizer | 0. 33c | 93.12 | 125 | 40.45 |
| Chemical agent | 1. 56b | 68.42 | 102 | 14.61 |
| Blank | 4. 94a | 89 |
Below the present invention is described in detail, the above, only the preferred embodiments of the invention, when can not limit
The scope of the present invention, that is, all according to the made impartial change of the application scope and modification, all should still belong in covering scope of the present invention.
Claims (3)
1. a kind of composite microbic bacterial fertilizer of preventing and treating graw mold of tomato is it is characterised in that the raw material of this composite microbic bacterial fertilizer is joined
Than for：Raw green streptomycete fermentation liquid 10-30 part, prosperous handkerchief watt bottom streptomycete fermentation liquid 20-40 part, India's fermentation of bacillus liquid
10-30 part, mocha husband's fermentation of bacillus liquid 10-30 part, ovum spore Trichoderma spp. fermentation liquid 10-30 part.
2. a kind of composite microbic bacterial fertilizer of preventing and treating graw mold of tomato according to claim 1 is it is characterised in that it is prepared
Method, comprises the following steps：
Step one：The preparation of raw green streptomycete fermentation liquid
Take out raw green streptomyces species preservation pipe, draw flat board with Gause I solid medium and recovered, cultivate 7 days for 30 DEG C,
Under flat board, the streak inoculation of picking single bacterium colony is equipped with the Fructus Solani melongenae bottle of 150 milliliters of Gause I solid mediums, in incubator
In 30 DEG C cultivate 6-8 days, treat that lawn covers with Fructus Solani melongenae bottle, produce a large amount of spore powders and can use 500 milliliters of physiological saline solution to wash
De-, regulation spore concentration is 0.1 hundred million cfu/ml, that is, green streptomycete seed liquor of making a living；
Fermentation：Green for the life of above-mentioned preparation streptomycete seed liquor is seeded to the green chain of life of the 600L equipped with sterilizing with 1% inoculum concentration
In the 1000L fermentation tank of mold fermentation culture medium, 28-32 DEG C of temperature control, open stirring 200r/min, first 24 hours, ventilation was every
Minute is 200L air, 24-36 hour, and ventilation is 400L, and after 36 hours, ventilation is 600L, cultivates 40-48 hour, treats
Thalline content reaches 60g/L, you can stop tank, that is, obtain raw green streptomycete fermentation liquid；
Wherein, described Gause I solid medium：Potassium nitrate：1 gram, soluble starch：20 grams, dipotassium hydrogen phosphate：0.5
Gram, magnesium sulfate：0.5 gram, sodium chloride：0.5 gram, ferrous sulfate：0.01 gram, agar：20 grams, 1000ml supplied by distilled water,
PH7.2～7.4；
Wherein, described life green streptomycete fermentation culture medium：Peanut meal powder 3%, glucose 1%, stone powder 1%, soy molasses 2%, phosphorus
Sour hydrogen dipotassium 0.04%, magnesium sulfate 0.05%, the condition of each medium sterilization is：0.10-0.15MPa, 121 DEG C sterilize 30 minutes；
Step 2：The preparation of prosperous handkerchief watt bottom streptomycete fermentation liquid
Take out prosperous handkerchief watt bottom streptomyces species preservation pipe, draw flat board with Gause I solid medium and recovered, 30 DEG C of cultures 7
My god, picking single bacterium colony streak inoculation under flat board, equipped with the Fructus Solani melongenae bottle of 150 milliliters of Gause I solid mediums, is being cultivated
Cultivate 6-8 days for 30 DEG C in case, treat that lawn covers with Fructus Solani melongenae bottle, produce the physiological saline solution that a large amount of spore powders can use 500 milliliters
Eluting, regulation spore concentration is 0.1 hundred million cfu/ml, as prosperous handkerchief watt bottom streptomycete seed liquor；
Fermentation：The prosperous handkerchief watt bottom streptomycete seed liquor of above-mentioned preparation is seeded to prosperous equipped with the 600L sterilizing with 1% inoculum concentration
In the 1000L fermentation tank of handkerchief watt bottom streptomycete fermentation culture medium, 28-30 DEG C of temperature control, open stirring 200r/min, first 18 hours, lead to
Tolerance be per minute for 300L air, 18-32 hour, ventilation is 500L, and after 32 hours, ventilation is 700L, cultivates 40-48
Hour, treat that thalline content reaches 80g/L, you can stop tank, that is, obtain prosperous handkerchief watt bottom streptomycete fermentation liquid；
Wherein, described Gause I solid medium with step 1 Gause I solid medium identical；
Wherein, described prosperous handkerchief watt bottom streptomycete fermentation culture medium：Sesame seed meal powder 3%, cottonseed meal 1%, corn starch 2%, stone powder 1%,
Cane molasses 3%, manganese sulfate 0.002%, dipotassium hydrogen phosphate 0.01%, magnesium sulfate 0.02%, the condition of each medium sterilization is：
0.10-0.15MPa, 121 DEG C sterilize 30 minutes；
Step 3, the preparation of India's fermentation of bacillus liquid
Take out India's bacillus cereuss preservation pipe, draw flat board respectively with LB solid medium and recovered, cultivate 48 hours for 30 DEG C,
Under flat board, picking single bacterium colony is seeded to equipped with LB solid medium, culture 48 hours in 30 DEG C of incubators, with 3000ml no
Bacterium water, by the lawn eluting in three Fructus Solani melongenae bottles, is seeded to the 500L fermentation tank equipped with 300L India fermentation of bacillus culture medium
In, open stirring 120r/min, ventilation is 200L/min within first 12 hours, after 12 hours, ventilation is 320L/min, 30 DEG C of cultures
26-36 hour, treats that total spore content is not less than 6,000,000,000 cfu/ml, you can as India's fermentation of bacillus liquid；
Wherein, described LB solid medium：Yeast extract 5.0 g, peptone 10.0, NaCl10.0 g, agar 20 g, water
1000mL, pH7.2；
Wherein, described India fermentation of bacillus culture medium：Glucose 8g/L, corn starch 22 g/L, bean cake powder 50 g/L,
Magnesium sulfate 0.4 g/L, manganese sulfate 0.5 g/L, potassium dihydrogen phosphate 0.5 g/L, Calcium Carbonate 5 g/L, pH7.0；Each medium sterilization
Condition be：0.10-0.15MPa, 121 DEG C sterilize 30 minutes；
Step 4, the preparation of mocha husband's fermentation of bacillus liquid
Take out mocha husband's bacillus cereuss preservation pipe, draw flat board respectively with nutrient broth solid medium and recovered, 30 DEG C of cultures
48 hours, under flat board, picking single bacterium colony was seeded to equipped with nutrient broth solid medium, cultivated 48 in 30 DEG C of incubators
Hour, with 3000ml sterilized water by the lawn eluting in three Fructus Solani melongenae bottles, it is seeded to equipped with 300L mocha husband's fermentation of bacillus
In the 500L fermentation tank of culture medium, open stirring 120r/min, ventilation is 200L/min within first 8 hours, and after 8 hours, ventilation is
300L/min, 30 DEG C of culture 24-36 hours, treat that total spore content is not less than 8,000,000,000 cfu/ml, you can as mocha husband's spore bar
Fermented liquid；
Wherein, described nutrient broth solid medium：Peptone 10.0 g/L, Carnis Bovis seu Bubali cream 3.0 g/L, sodium chloride 5.0 g/L,
Agar 20 g/L, pH 7.2 ± 0.2；
Wherein, described mocha husband's fermentation of bacillus culture medium：Glucose 8g/L, tapioca starch 20 g/L, rapeseed cake powder 30 g/L,
Magnesium sulfate 0.4 g/L, manganese sulfate 0.5 g/L, potassium dihydrogen phosphate 0.2 g/L, Calcium Carbonate 5 g/L, pH7.0；Each medium sterilization
Condition be：0.10-0.15MPa, 121 DEG C sterilize 30 minutes；
Step 5, the preparation of ovum spore Trichoderma spp. fermentation liquid
Take out ovum spore Trichoderma spp. strain preservation pipe, draw flat board with PDA solid medium and recovered, cultivate 7 days, under flat board for 30 DEG C
Picking single bacterium colony streak inoculation, equipped with the Fructus Solani melongenae bottle of 150 milliliters of PDA solid mediums, cultivates 6-8 for 30 DEG C in incubator
My god, treat that lawn covers with Fructus Solani melongenae bottle, produce the physiological saline solution eluting that a large amount of spore powders can use 500 milliliters, adjust spore dense
Spend for 0.05 hundred million cfu/ml, as ovum spore Trichoderma spp. seed liquor；
Fermentation：The ovum spore Trichoderma spp. seed liquor of above-mentioned preparation is seeded to equipped with 300L ovum spore Trichoderma spp. fermentation culture with 2% inoculum concentration
In the 500L fermentation tank of base, open stirring 200r/min, ventilation is 120L/min within first 8 hours, and after 8-24 hour, ventilation is
240L/min, after 24 hours, ventilation is 360L/min, 30 DEG C of culture 32-48 hours, treats that thalline content reaches 80g/L, that is,
Tank can be stopped, you can as ovum spore Trichoderma spp. fermentation liquid；
Wherein, described PDA solid medium：Rhizoma Solani tuber osi 200g, sucrose 20g, water 1000mL, agar 20g；
Wherein, described ovum spore Trichoderma spp. fermentation medium：Rapeseed cake 2%, cottonseed meal 2%, sweet potato flour 2%, stone powder 1%, soy molasses 3%, sulfur
Sour manganese 0.001%, dipotassium hydrogen phosphate 0.04%, magnesium sulfate 0.02%, the condition of each medium sterilization is：0.10-0.15MPa, 121
DEG C sterilizing 30 minutes；
Step 6, mixed fermentation liquid
The bacterium solution of first five step preparation is pressed raw green streptomycete fermentation liquid 10-30 part, prosperous handkerchief watt bottom streptomycete fermentation liquid 20-40
Part, India's fermentation of bacillus liquid 10-30 part, mocha husband's fermentation of bacillus liquid 10-30 part, ovum spore Trichoderma spp. fermentation liquid 10-30
Part；Mix, that is, obtain preventing and treating the composite microbic bacterial fertilizer of graw mold of tomato.
3. a kind of composite microbic bacterial fertilizer of preventing and treating graw mold of tomato according to claim 1 is it is characterised in that this is compound
Microbial-bacterial fertilizer is foliage-spray each 2-5L composite microbic bacterial fertilizer/mu in the method for preventing and treating graw mold of tomato, and it uses dilute
The multiple released is 100-200 times, continuously uses 2-3 time, every 7-10 days using once.
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| CN108142247A (en) * | 2017-12-23 | 2018-06-12 | 青岛麦迪尔生物工程有限公司 | A kind of biological and ecological methods to prevent plant disease, pests, and erosion culture medium for cultivating matter of anti-graw mold of tomato |
| CN108203703A (en) * | 2018-03-26 | 2018-06-26 | 叶志坚 | A kind of biocontrol agent and its application in gray mold is prevented |
| CN108293480A (en) * | 2018-03-26 | 2018-07-20 | 叶志坚 | The method for preventing graw mold of tomato using biocontrol agent |
| CN108624527A (en) * | 2018-05-12 | 2018-10-09 | 湖南科技学院 | A kind of ginkgo source growth-promoting preparation of prevention ginger bacterial wilt |
| CN114410518A (en) * | 2022-01-07 | 2022-04-29 | 春华秋实科技集团有限公司 | Compound microbial agent containing streptomyces and application thereof |
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| CN105199996A (en) * | 2015-11-03 | 2015-12-30 | 河北省农林科学院植物保护研究所 | Bacillus amyloliquefaciens for controlling tomato gray mould and application of bacillus amyloliquefaciens |
| CN105777329A (en) * | 2016-02-04 | 2016-07-20 | 鞍山禾瑞生物科技有限公司 | Solid composite biological multi-control water and moisture retention fertilizer and preparing method and application thereof |
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| CN108142247A (en) * | 2017-12-23 | 2018-06-12 | 青岛麦迪尔生物工程有限公司 | A kind of biological and ecological methods to prevent plant disease, pests, and erosion culture medium for cultivating matter of anti-graw mold of tomato |
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| CN108293480A (en) * | 2018-03-26 | 2018-07-20 | 叶志坚 | The method for preventing graw mold of tomato using biocontrol agent |
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| CN108624527A (en) * | 2018-05-12 | 2018-10-09 | 湖南科技学院 | A kind of ginkgo source growth-promoting preparation of prevention ginger bacterial wilt |
| CN114410518A (en) * | 2022-01-07 | 2022-04-29 | 春华秋实科技集团有限公司 | Compound microbial agent containing streptomyces and application thereof |
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