CN103146600A - Antagonistic bacteria for prevention and treatment of tobacco bacterial wilt and application thereof - Google Patents
Antagonistic bacteria for prevention and treatment of tobacco bacterial wilt and application thereof Download PDFInfo
- Publication number
- CN103146600A CN103146600A CN2013100461713A CN201310046171A CN103146600A CN 103146600 A CN103146600 A CN 103146600A CN 2013100461713 A CN2013100461713 A CN 2013100461713A CN 201310046171 A CN201310046171 A CN 201310046171A CN 103146600 A CN103146600 A CN 103146600A
- Authority
- CN
- China
- Prior art keywords
- bacterial wilt
- tobacco
- preservation
- tobacco bacterial
- antagonistic fungi
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Granted
Links
- 241000208125 Nicotiana Species 0.000 title claims abstract description 62
- 235000002637 Nicotiana tabacum Nutrition 0.000 title claims abstract description 62
- 230000001580 bacterial effect Effects 0.000 title claims abstract description 51
- 230000003042 antagnostic effect Effects 0.000 title claims abstract description 46
- 241000894006 Bacteria Species 0.000 title abstract description 40
- 230000002265 prevention Effects 0.000 title abstract description 5
- 238000011282 treatment Methods 0.000 title abstract description 5
- 239000003895 organic fertilizer Substances 0.000 claims abstract description 18
- 238000004321 preservation Methods 0.000 claims abstract description 18
- 230000000813 microbial effect Effects 0.000 claims abstract description 17
- 241000228245 Aspergillus niger Species 0.000 claims abstract description 12
- 241000193755 Bacillus cereus Species 0.000 claims abstract description 12
- 239000003795 chemical substances by application Substances 0.000 claims abstract description 12
- 241000233866 Fungi Species 0.000 claims description 33
- 235000016709 nutrition Nutrition 0.000 abstract description 2
- 230000035764 nutrition Effects 0.000 abstract description 2
- 239000011368 organic material Substances 0.000 abstract 1
- 239000002689 soil Substances 0.000 description 36
- 201000010099 disease Diseases 0.000 description 22
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 22
- 235000019504 cigarettes Nutrition 0.000 description 18
- 244000005700 microbiome Species 0.000 description 17
- 239000002609 medium Substances 0.000 description 15
- 244000052616 bacterial pathogen Species 0.000 description 14
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 13
- 238000000034 method Methods 0.000 description 11
- 239000000725 suspension Substances 0.000 description 11
- 230000001954 sterilising effect Effects 0.000 description 9
- 229920001817 Agar Polymers 0.000 description 8
- 239000008272 agar Substances 0.000 description 8
- 210000003608 fece Anatomy 0.000 description 8
- 238000004659 sterilization and disinfection Methods 0.000 description 8
- 241000196324 Embryophyta Species 0.000 description 7
- 230000000694 effects Effects 0.000 description 7
- 239000010871 livestock manure Substances 0.000 description 7
- 238000004519 manufacturing process Methods 0.000 description 7
- 238000000926 separation method Methods 0.000 description 7
- 239000000243 solution Substances 0.000 description 7
- 239000001888 Peptone Substances 0.000 description 6
- 239000007788 liquid Substances 0.000 description 6
- 239000002068 microbial inoculum Substances 0.000 description 6
- 235000015097 nutrients Nutrition 0.000 description 6
- IICCLYANAQEHCI-UHFFFAOYSA-N 4,5,6,7-tetrachloro-3',6'-dihydroxy-2',4',5',7'-tetraiodospiro[2-benzofuran-3,9'-xanthene]-1-one Chemical compound O1C(=O)C(C(=C(Cl)C(Cl)=C2Cl)Cl)=C2C21C1=CC(I)=C(O)C(I)=C1OC1=C(I)C(O)=C(I)C=C21 IICCLYANAQEHCI-UHFFFAOYSA-N 0.000 description 5
- 230000000844 anti-bacterial effect Effects 0.000 description 5
- 238000002474 experimental method Methods 0.000 description 5
- 238000000855 fermentation Methods 0.000 description 5
- 230000004151 fermentation Effects 0.000 description 5
- 239000003337 fertilizer Substances 0.000 description 5
- 239000002054 inoculum Substances 0.000 description 5
- 239000000203 mixture Substances 0.000 description 5
- 230000008569 process Effects 0.000 description 5
- 229930187593 rose bengal Natural products 0.000 description 5
- 229940081623 rose bengal Drugs 0.000 description 5
- STRXNPAVPKGJQR-UHFFFAOYSA-N rose bengal A Natural products O1C(=O)C(C(=CC=C2Cl)Cl)=C2C21C1=CC(I)=C(O)C(I)=C1OC1=C(I)C(O)=C(I)C=C21 STRXNPAVPKGJQR-UHFFFAOYSA-N 0.000 description 5
- 238000012216 screening Methods 0.000 description 5
- 238000012360 testing method Methods 0.000 description 5
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 description 4
- 238000004458 analytical method Methods 0.000 description 4
- 235000015278 beef Nutrition 0.000 description 4
- 230000009514 concussion Effects 0.000 description 4
- 238000011081 inoculation Methods 0.000 description 4
- 230000001717 pathogenic effect Effects 0.000 description 4
- 238000002360 preparation method Methods 0.000 description 4
- 230000002792 vascular Effects 0.000 description 4
- 235000015099 wheat brans Nutrition 0.000 description 4
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 description 3
- 241000589771 Ralstonia solanacearum Species 0.000 description 3
- 206010039509 Scab Diseases 0.000 description 3
- 229920002472 Starch Polymers 0.000 description 3
- 230000008485 antagonism Effects 0.000 description 3
- 239000011248 coating agent Substances 0.000 description 3
- 238000000576 coating method Methods 0.000 description 3
- 206010025482 malaise Diseases 0.000 description 3
- 238000012163 sequencing technique Methods 0.000 description 3
- 239000008107 starch Substances 0.000 description 3
- 235000019698 starch Nutrition 0.000 description 3
- FRXSZNDVFUDTIR-UHFFFAOYSA-N 6-methoxy-1,2,3,4-tetrahydroquinoline Chemical compound N1CCCC2=CC(OC)=CC=C21 FRXSZNDVFUDTIR-UHFFFAOYSA-N 0.000 description 2
- 241000186361 Actinobacteria <class> Species 0.000 description 2
- 241000726221 Gemma Species 0.000 description 2
- 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 description 2
- 238000003794 Gram staining Methods 0.000 description 2
- 108010080698 Peptones Proteins 0.000 description 2
- 241001002356 Valeriana edulis Species 0.000 description 2
- 230000009286 beneficial effect Effects 0.000 description 2
- WIIZWVCIJKGZOK-RKDXNWHRSA-N chloramphenicol Chemical compound ClC(Cl)C(=O)N[C@H](CO)[C@H](O)C1=CC=C([N+]([O-])=O)C=C1 WIIZWVCIJKGZOK-RKDXNWHRSA-N 0.000 description 2
- 235000019219 chocolate Nutrition 0.000 description 2
- 230000006378 damage Effects 0.000 description 2
- 238000010790 dilution Methods 0.000 description 2
- 239000012895 dilution Substances 0.000 description 2
- 238000005516 engineering process Methods 0.000 description 2
- 238000011156 evaluation Methods 0.000 description 2
- 238000011049 filling Methods 0.000 description 2
- 239000012467 final product Substances 0.000 description 2
- 239000012634 fragment Substances 0.000 description 2
- 239000008103 glucose Substances 0.000 description 2
- 239000001963 growth medium Substances 0.000 description 2
- 230000036541 health Effects 0.000 description 2
- 230000005764 inhibitory process Effects 0.000 description 2
- 238000011835 investigation Methods 0.000 description 2
- 230000007246 mechanism Effects 0.000 description 2
- 230000000877 morphologic effect Effects 0.000 description 2
- 229940049547 paraxin Drugs 0.000 description 2
- 235000019319 peptone Nutrition 0.000 description 2
- 239000000575 pesticide Substances 0.000 description 2
- 239000006041 probiotic Substances 0.000 description 2
- 235000018291 probiotics Nutrition 0.000 description 2
- 108090000623 proteins and genes Proteins 0.000 description 2
- 238000011218 seed culture Methods 0.000 description 2
- 239000011780 sodium chloride Substances 0.000 description 2
- 239000000126 substance Substances 0.000 description 2
- 108020004465 16S ribosomal RNA Proteins 0.000 description 1
- 108020004463 18S ribosomal RNA Proteins 0.000 description 1
- 108020004565 5.8S Ribosomal RNA Proteins 0.000 description 1
- GUBGYTABKSRVRQ-XLOQQCSPSA-N Alpha-Lactose Chemical compound O[C@@H]1[C@@H](O)[C@@H](O)[C@@H](CO)O[C@H]1O[C@@H]1[C@@H](CO)O[C@H](O)[C@H](O)[C@H]1O GUBGYTABKSRVRQ-XLOQQCSPSA-N 0.000 description 1
- 241000193830 Bacillus <bacterium> Species 0.000 description 1
- 102000016938 Catalase Human genes 0.000 description 1
- 108010053835 Catalase Proteins 0.000 description 1
- KRKNYBCHXYNGOX-UHFFFAOYSA-K Citrate Chemical compound [O-]C(=O)CC(O)(CC([O-])=O)C([O-])=O KRKNYBCHXYNGOX-UHFFFAOYSA-K 0.000 description 1
- FBPFZTCFMRRESA-KVTDHHQDSA-N D-Mannitol Chemical compound OC[C@@H](O)[C@@H](O)[C@H](O)[C@H](O)CO FBPFZTCFMRRESA-KVTDHHQDSA-N 0.000 description 1
- 108010010803 Gelatin Proteins 0.000 description 1
- SRBFZHDQGSBBOR-HWQSCIPKSA-N L-arabinopyranose Chemical compound O[C@H]1COC(O)[C@H](O)[C@H]1O SRBFZHDQGSBBOR-HWQSCIPKSA-N 0.000 description 1
- GUBGYTABKSRVRQ-QKKXKWKRSA-N Lactose Natural products OC[C@H]1O[C@@H](O[C@H]2[C@H](O)[C@@H](O)C(O)O[C@@H]2CO)[C@H](O)[C@@H](O)[C@H]1O GUBGYTABKSRVRQ-QKKXKWKRSA-N 0.000 description 1
- 229930195725 Mannitol Natural products 0.000 description 1
- 241001465754 Metazoa Species 0.000 description 1
- 229910002651 NO3 Inorganic materials 0.000 description 1
- 244000061176 Nicotiana tabacum Species 0.000 description 1
- NHNBFGGVMKEFGY-UHFFFAOYSA-N Nitrate Chemical compound [O-][N+]([O-])=O NHNBFGGVMKEFGY-UHFFFAOYSA-N 0.000 description 1
- 241000237502 Ostreidae Species 0.000 description 1
- OAICVXFJPJFONN-UHFFFAOYSA-N Phosphorus Chemical compound [P] OAICVXFJPJFONN-UHFFFAOYSA-N 0.000 description 1
- ZLMJMSJWJFRBEC-UHFFFAOYSA-N Potassium Chemical compound [K] ZLMJMSJWJFRBEC-UHFFFAOYSA-N 0.000 description 1
- 108010009736 Protein Hydrolysates Proteins 0.000 description 1
- 240000004808 Saccharomyces cerevisiae Species 0.000 description 1
- 229930006000 Sucrose Natural products 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
- 239000002253 acid Substances 0.000 description 1
- SRBFZHDQGSBBOR-LECHCGJUSA-N alpha-D-xylose Chemical compound O[C@@H]1CO[C@H](O)[C@H](O)[C@H]1O SRBFZHDQGSBBOR-LECHCGJUSA-N 0.000 description 1
- SRBFZHDQGSBBOR-UHFFFAOYSA-N beta-D-Pyranose-Lyxose Natural products OC1COC(O)C(O)C1O SRBFZHDQGSBBOR-UHFFFAOYSA-N 0.000 description 1
- 230000000443 biocontrol Effects 0.000 description 1
- 238000009835 boiling Methods 0.000 description 1
- 229940041514 candida albicans extract Drugs 0.000 description 1
- 239000002775 capsule Substances 0.000 description 1
- 239000005018 casein Substances 0.000 description 1
- BECPQYXYKAMYBN-UHFFFAOYSA-N casein, tech. Chemical compound NCCCCC(C(O)=O)N=C(O)C(CC(O)=O)N=C(O)C(CCC(O)=N)N=C(O)C(CC(C)C)N=C(O)C(CCC(O)=O)N=C(O)C(CC(O)=O)N=C(O)C(CCC(O)=O)N=C(O)C(C(C)O)N=C(O)C(CCC(O)=N)N=C(O)C(CCC(O)=N)N=C(O)C(CCC(O)=N)N=C(O)C(CCC(O)=O)N=C(O)C(CCC(O)=O)N=C(O)C(COP(O)(O)=O)N=C(O)C(CCC(O)=N)N=C(O)C(N)CC1=CC=CC=C1 BECPQYXYKAMYBN-UHFFFAOYSA-N 0.000 description 1
- 235000021240 caseins Nutrition 0.000 description 1
- 230000001364 causal effect Effects 0.000 description 1
- 230000001413 cellular effect Effects 0.000 description 1
- 239000002361 compost Substances 0.000 description 1
- 238000003967 crop rotation Methods 0.000 description 1
- 238000000354 decomposition reaction Methods 0.000 description 1
- 238000013461 design Methods 0.000 description 1
- 230000001066 destructive effect Effects 0.000 description 1
- 238000007865 diluting Methods 0.000 description 1
- 239000012153 distilled water Substances 0.000 description 1
- 230000003203 everyday effect Effects 0.000 description 1
- 239000004744 fabric Substances 0.000 description 1
- 230000035558 fertility Effects 0.000 description 1
- 230000002538 fungal effect Effects 0.000 description 1
- 239000008273 gelatin Substances 0.000 description 1
- 229920000159 gelatin Polymers 0.000 description 1
- 235000019322 gelatine Nutrition 0.000 description 1
- 235000011852 gelatine desserts Nutrition 0.000 description 1
- 101150013736 gyrB gene Proteins 0.000 description 1
- 230000007062 hydrolysis Effects 0.000 description 1
- 238000006460 hydrolysis reaction Methods 0.000 description 1
- 238000011534 incubation Methods 0.000 description 1
- 230000002262 irrigation Effects 0.000 description 1
- 238000003973 irrigation Methods 0.000 description 1
- 239000003621 irrigation water Substances 0.000 description 1
- 238000002955 isolation Methods 0.000 description 1
- 239000008101 lactose Substances 0.000 description 1
- 244000144972 livestock Species 0.000 description 1
- 239000000594 mannitol Substances 0.000 description 1
- 235000010355 mannitol Nutrition 0.000 description 1
- 239000000463 material Substances 0.000 description 1
- 239000012528 membrane Substances 0.000 description 1
- 239000002207 metabolite Substances 0.000 description 1
- CEQFOVLGLXCDCX-WUKNDPDISA-N methyl red Chemical compound C1=CC(N(C)C)=CC=C1\N=N\C1=CC=CC=C1C(O)=O CEQFOVLGLXCDCX-WUKNDPDISA-N 0.000 description 1
- 235000020636 oyster Nutrition 0.000 description 1
- 238000012856 packing Methods 0.000 description 1
- 235000011837 pasties Nutrition 0.000 description 1
- 239000011574 phosphorus Substances 0.000 description 1
- 229910052698 phosphorus Inorganic materials 0.000 description 1
- 239000000049 pigment Substances 0.000 description 1
- 239000011591 potassium Substances 0.000 description 1
- 229910052700 potassium Inorganic materials 0.000 description 1
- 244000144977 poultry Species 0.000 description 1
- 238000012545 processing Methods 0.000 description 1
- 238000012797 qualification Methods 0.000 description 1
- 108700022487 rRNA Genes Proteins 0.000 description 1
- 230000009467 reduction Effects 0.000 description 1
- 230000001105 regulatory effect Effects 0.000 description 1
- 239000004576 sand Substances 0.000 description 1
- 239000000779 smoke Substances 0.000 description 1
- 241000894007 species Species 0.000 description 1
- 238000003756 stirring Methods 0.000 description 1
- 239000005720 sucrose Substances 0.000 description 1
- 208000024891 symptom Diseases 0.000 description 1
- 238000012546 transfer Methods 0.000 description 1
- 238000002054 transplantation Methods 0.000 description 1
- 230000003442 weekly effect Effects 0.000 description 1
- 229960003487 xylose Drugs 0.000 description 1
- 239000012138 yeast extract Substances 0.000 description 1
Images
Abstract
The invention discloses antagonistic bacteria for prevention and treatment of tobacco bacterial wilt and application thereof. The antagonistic bacteria are Bacillus cereus QJ-1 and Aspergillus niger Ty-3. Specifically, the preservation unit of QJ-1 is China Center for Type Culture Collection, the preservation address is Wuhan University, Wuhan, China, the preservation date is July 5, 2012, and the preservation number is: CCTCC, NO:M2012271; and the preservation unit of Ty-3 is China Center for Type Culture Collection, the preservation address is Wuhan University, Wuhan, China, the preservation date is July 8, 2011, and the preservation number is CCTCC NO:M2011241. The antagonistic bacteria, i.e. the Bacillus cereus QJ-1 and Aspergillus niger Ty-3 provided in the invention can be combined into a microbial agent. The microbial agent and an organic material can be fermented into a microbial organic fertilizer, which can provide effective nutrition and functional bacteria, and realize effective prevention and treatment of tobacco bacterial wilt.
Description
Technical field
The present invention relates to prevent and treat the tobacco bacterial wilt Antagonistic Fungi, also relate to simultaneously this Antagonistic Fungi in the application of control aspect soil-borne disease of tobacco, belong to the biological control field.
Background technology
Tobacco bacterial wilt is the important soil-borne disease of tobacco of a class, and it infects the tobacco root usually, causes the disease of crop root and even complete stool, causes great financial loss.Tobacco bacterial wilt is that a kind of systematicness that is caused by cloth Ke Shi bacillus (Ralstonia solanacearum) infects disease, and in a single day the cigarette strain is caught an illness and often caused whole strain dead, and its harm is destructive often, therefore usually produces to tobacco and causes heavy economic losses.The tobacco bacterial wilt pathogenic bacteria is mainly lost to fall within soil with plant residue and survives the winter, also can survive the winter in seed or in other pin main body of field, the soil that carries disease germs, invalid tissue and the organic fertilizer etc. that contains germ are the main sources of just dying of this disease, and the propagation of disease is mainly by irrigation water, rainwater, seedling, farm implements, sick soil and people and animals' activity etc. in spite of illness.
At present, in production, the main agricultural measures such as disease-resistant variety and shift of crops that adopt are prevented and treated the soil-borne disease of tobacco disease, but because disease-resistant variety needs the disease-resistant gene variation, and being subjected to the impact of environment larger, prevention effect is undesirable.Shift of crops is also one of effective ways of control tobacco bacterial wilt, but because China has a large population and a few land, in production, normal crop rotation measure can't realize.The phytopathologist thinks that plant morbidity and these three factors of host, pathogenic bacteria and environment have substantial connection, when pathogenic bacteria and susceptible host meet in adapt circumstance, plant just begins morbidity, if revise or eradicate any one in this three, just can alleviate or control Plant diseases.
Studies show that in a large number, not only exist in vega soil to cause the microorganism of tobacco diseases but also to have beneficial microorganism diversified non-virulent, that improve tobacco vitality, thereby the biological control of tobacco and bionomic control come into one's own day by day.Bio-control method is to utilize beneficial microorganism to reduce phytopathogen to the harm of plant, bionomic control is by measures such as species diversities in soil introducing antagonistic microbe, fertilizing soil and raising soil, and then make soil micro-ecosystem reach balance, finally by the antagonism between different biologies in soil and the control of competition realization to soil disease.
Summary of the invention
The purpose of this invention is to provide control tobacco bacterial wilt Antagonistic Fungi.
In order to realize above purpose, the technical solution adopted in the present invention is to provide control tobacco bacterial wilt Antagonistic Fungi, and described Antagonistic Fungi is bacillus cereus (Bacillus cereus) QJ-1 and aspergillus niger (Aspergillusniger) Ty-3; Wherein, bacillus cereus (Bacillus cereus) QJ-1, depositary institution: Chinese Typical Representative culture collection center, preservation address: Wuhan, China Wuhan University, preservation date: on July 5th, 2012, preserving number: CCTCC NO:M2012271; Aspergillus niger (Aspergillus niger) Ty-3, depositary institution: Chinese Typical Representative culture collection center, preservation address: Wuhan, China Wuhan University, preservation date: on July 8th, 2011, preserving number: CCTCC NO:M2011241.
The present invention prevents and treats the screening method of tobacco bacterial wilt Antagonistic Fungi, comprises the following steps:
1) pathogenicbacteria separation
Choose the cigarette strain of tobacco bacterial wilt morbidity, tobacco rod is cleaned, sterilizes, get scab on tobacco rod, susceptible position vascular bundle is organized put into that the beef extract-peptone bacterium is dull and stereotyped to be cultivated after stripping and slicing; When Stalk Rot was grown to mycelia, picking colony purifying, cultivation were inoculated in beef extract-peptone inclined-plane preservation;
2) Pathogenic is identified
Sow tobacco seed, when the cigarette strain has grown to vascular bundle and organizes, destroy the tobacco root and also pour the bacteria suspension of cultured Stalk Rot into root, keep hot and humid, observe incidence after 30d;
3) separation of soil microorganisms
Get the cigarette strain rhizosphere soil on every side of catching an illness, add sterilized water, standing after concussion, the soil solution is carried out the concentration gradient dilution, get diluting soln evenly coating on beef-protein medium, Gause I substratum and rose bengal medium respectively, cultivate, picking list bacterium colony purifying also is seeded to corresponding slant medium preservation;
4) Antagonistic Fungi screening
With the cultivation that stands facing each other on the NA substratum of the pathogenic bacteria of the different soils microorganism cultivated and bacterial wilt, judge to have or not antagonistic action;
5) antagonistic action is measured
Wash out Stalk Rot with sterilized water, the preparation bacteria suspension, and bacteria suspension is mixed with the NA substratum, placement is frozen into flat board, single and the combined inoculation in twos of the soil microorganisms with antagonistic action measure is cultivated in face-off cultivate on flat board, the mensuration soil microorganisms is to the antibacterial spot of pathogenic bacteria or the size of antibacterial band;
6) Antagonistic Fungi is carried out morphological observation;
7) Antagonistic Fungi is identified classification.
The present invention also aims to provide the application of control tobacco bacterial wilt Antagonistic Fungi aspect microbiobacterial agent.
The technical solution adopted in the present invention also is to provide the application of control tobacco bacterial wilt Antagonistic Fungi aspect microbiobacterial agent.
the present invention prevents and treats the production method of tobacco bacterial wilt Antagonistic Fungi microbial inoculum: Antagonistic Fungi QJ-1 and the Ty-3 of purifying are inoculated into respectively on the NA substratum, cultivate 24h in the incubator of 32 ℃, get 5mL sterilized water contra bevel, vibration gently, then accurately drawing the lmL bacteria suspension joins in the 250mL triangular flask that the aseptic NA liquid nutrient medium of 150mL is housed, 1d is cultivated in the shaking bath concussion that is put into 32 ℃, then the inoculum size of cultured bacteria suspension according to massfraction 10% joined respectively in the wheat bran of sterilization, cultivate 3d for 32 ℃, the air-dry single bacteria agent that is QJ-1 and Ty-3.In single bacteria agent, QJ-1 and Ty-3 reach respectively 10
8Then cfu/g mixes two kinds of single bacteria agents in the ratio of quality 1:1, be the Antagonistic Fungi microbial inoculum.
The present invention also aims to provide the application of control tobacco bacterial wilt Antagonistic Fungi aspect microbial organic fertilizer.
The technical solution adopted in the present invention also is to provide the application of control tobacco bacterial wilt Antagonistic Fungi aspect microbial organic fertilizer.
The production method of control tobacco bacterial wilt Antagonistic Fungi microbial organic fertilizer of the present invention: the addition by massfraction 8～12% adds microbial inoculum decomposed manure to mix equably, regulate moisture to 50～60%, pile the stacking of 50～80cm, 5～7d and get final product banks up.
The present invention sets about from improving the soil microorganisms aspect, from original position soil screening Antagonistic Fungi, and has studied the application of control tobacco bacterial wilt Antagonistic Fungi in microbiobacterial agent and microbial organic fertilizer.Antagonistic Fungi mixes the microorganism organic fertilizer that makes with organic fertilizer can significantly reduce tobacco bacterial wilt, and this fertilizer provides effective nutrition and function yeast, and Antagonistic Fungi is grown rapidly in soil surely, regulates the soil micro-ecosystem balance and suppresses bacterial wilt.Can reach by use this functional fertilizer to vega the ability that bacterial wilt is resisted in cigarette strain self that improves, improve the purpose of quality of tobacco and minimizing chemical fertilizer, pesticide dosage.
Description of drawings
Fig. 1 is for separating the gramstaining of the bacterial isolates QJ-1 that obtains from original position soil;
Fig. 2 is for separating the fungal bacterial strain Ty-3 that obtains from original position soil;
Fig. 3 is that QJ-1 and Ty-3 mix and ralstonia solanacearum face-off growth 1 day;
Fig. 4 is the microscopic morphology figure of Ty-3 spore.
Embodiment
The tobacco bacterial wilt pathogenic bacteria is taken from Shaowu City, Fujian Province cigarette district and has the cigarette strain of more typical illness, and gets the soil around the diseased plant rhizosphere.
The preparation of substratum:
Beef-protein medium: NaCl 5g, extractum carnis 3g, peptone 10g, agar 20g adds water and is settled to 1L, pH7.4～7.6,121 ℃ sterilization 20min.
Gause I substratum: Zulkovsky starch 20g, KNO
31g, K
2HPO
40.5g, MgSO
47H
2O 0.5g, NaCl 0.5g, FeSO
47H
2O 0.01g, agar 20g, pH7.4～7.6.During preparation, first use a small amount of cold water, with starch furnishing pasty state, pour in the boiling water that is less than institute's water requirement, heat on fire, dissolve one by one successively while stirring other compositions, after dissolving, supply moisture to 1000mL, transfer pH, 121 ℃ of sterilization 20min.
Rose bengal medium: peptone 5g, glucose 10g, KH
2PO
41g, MgSO
47H
2O 0.5g, agar 20g, 1/3000 rose-bengal solution 100mL, paraxin 0.1g.After above-mentioned each composition adds in distilled water dissolving, then add rose-bengal solution.Separately with a small amount of dissolve with ethanol paraxin, add in substratum, after packing, 121 ℃ of sterilization 20min.
NA substratum: yeast extract 12g, sucrose 20g, K
2HPO
44g, agar 20g, pH7.0～7.2 add the water constant volume to 1L, 121 ℃ of sterilization 20min.The difference of NA liquid nutrient medium and NA substratum is, the NA liquid nutrient medium does not add agar.
Wort agar substratum (MEA): wort 150mL, agar 3g, pH nature (approximately 6.4), 121 ℃ of sterilization 20min.
The isolation and screening of embodiment 1, tobacco bacterial wilt Antagonistic Fungi
1) pathogenicbacteria separation
Before separating germ, first tobacco rod is rinsed well, with 75% alcohol, tobacco rod and pocket knife are sterilized again, then cut scab on cigarette strain cane open with pocket knife under aseptic condition, susceptible position vascular bundle tissue is cut into some fritters, put into beef extract-peptone bacterium flat board culture dish with the tweezers gripping cigarette piece of sterilizing and cultivate, put 5 in every ware, repeat 3 wares, culture temperature is 32 ℃, and incubation time is 48h.Treat that bacterial wilt cigarette piece and substratum contact position grow white mobility bacterium colony and outwards disperse tiny thalline, be inoculated in beef extract-peptone and preserve.
Photomicrograph shows that the Stalk Rot thalline is shaft-like, the blunt circle in two ends, and amphitrichous, without pod membrane, Gram-negative.
2) pathogenic evaluation
Select tobacco K326 to be the test kind.With tobacco seed after planting, when the cigarette strain has grown into vascular bundle and organizes, the root of tobacco destroyed and cultured Stalk Rot weaker concn gradient is made bacteria suspension in the basin alms bowl, pouring the 50mL bacteria suspension in the tobacco rhizosphere, and keep hot and humid.Observe incidence after 30d.
According to the KochShi rule, the bacterial wilt pathogenic strains of separating in the cigarette strain of falling ill is inoculated on the health tobacco plant, pathogenic to measure it.With isolated Causal Organism of The Bacterial Wilt strains tested inoculation 20 strain health tobaccos, there are 15 strain tobaccos the bacterial wilt symptom to occur through after 30d.
3) separation of soil microorganisms
Obtain bacterial wilt cigarette strain rhizosphere soil 10g on every side, put into the triangular flask of 90mL sterilized water, then standing 20min after concussion 30min is diluted to 10
-4, 10
-5With 10
-6G/mL, draw respectively solution 0.1mL evenly coating on beef-protein medium, Gause I substratum and rose bengal medium of different concns, every kind is repeated 3 wares, and after being placed in 32 ℃ of incubators cultivation 48h, picking list bacterium colony carries out purifying and move on corresponding slant medium preserving.
Separate through dull and stereotyped dilution spread, obtain altogether 5 kinds of bacteriums, 4 kinds of fungies and 4 kinds of actinomycetes from soil.Bacterium is numbered respectively QJ-1 to QJ-5, and fungi is numbered Ty-1 to Ty-4, and actinomycetes are numbered QF-1 to QF-4.In bacterium, QJ-1 is gram-positive microorganism, and all the other 4 strain bacterium are Gram-negative bacteria, and QJ-1 gramstaining photo as shown in Figure 1.The colonial morphology figure of Ty-3 as shown in Figure 2.
4) screening of antagonistic strain
The cultivation that stands facing each other on the NA substratum of the soil microorganisms that separation is obtained and Stalk Rot.Substratum coating Stalk Rot, middle inoculation separates the soil microorganisms that obtains, two bacterium are 3mm apart, check cultivation results after 2d, according to having or not inhibition zone etc. to judge that the soil microorganisms of separation has or not antagonistic action to pathogenic bacteria between bacterium colony tempo, bacterium colony.
Cultivate by face-off, filter out 3 strains and the tobacco bacterial wilt germ is had the bacterial strain of antagonistic action from the soil microorganisms that separation obtains, be respectively QJ-1, QJ-3 and Ty-3.
5) antagonistic action is measured
wash out the pathogenic bacteria of the bacterial wilt of cultivating 24h with sterilized water, make bacteria suspension, draw in the culture dish of bacteria suspension after sterilization of 1mL, then pour into 45-50 ℃ NA substratum 15mL, place 2h and solidify rear one-tenth flat board, single and the combined inoculation in twos of the soil microorganisms with antagonistic action measure is cultivated in face-off cultivates on flat board, every kind is repeated 3 wares, observe the growing state of pathogenic bacteria after cultivating 2d under 32 ℃, and measure soil microorganisms to the antibacterial spot of germ or the size of antibacterial band, wherein the antibacterial band of QJ-1 and Ty-3 mixed bacterium is maximum, reach 9mm, has the effect that suppresses preferably Stalk Rot, as shown in Figure 3.
6) antagonism mechanism
QJ-1 and the Ty-3 antagonism mechanism to the tobacco bacterial wilt germ:
QJ-1 and Ty-3 can produce some meta-bolites, and this substance effectively suppresses the growth of Stalk Rot, and both mix the generation that mutual promotion suppresses the pathogenic bacteria material, strengthen bacteriostasis.
Embodiment 2, morphological observation
QJ-1 bacterium initial stage bacterium colony is open and flat, and oyster white is smooth, and is opaque.
Ty-3 bacterium initial stage white hypha, rear generation dark-brown or chocolate spore, spore is abundant.
The evaluation classification of embodiment 3, Antagonistic Fungi
Qualification result is through institute of microbiology of the Chinese Academy of Sciences:
1, QJ-1 is bacillus cereus (Bacillus cereus).
1) cellular form and physics and chemistry test-results
Gram-positive, cell are shaft-like, and cell dia forms gemma greater than 1 μ m, and gemma does not expand; VP test, methyl red test, Starch Hydrolysis, decomposition casein, gelatin hydrolysate, catalase, oxydase, nitrate reduction react, utilize Citrate trianion all to be positive; Acid-producing shows as: glucose is positive, wood sugar, L-arabinose, N.F,USP MANNITOL, lactose feminine gender.
2) 16S rRNA gene sequencing result
GCAGTCGAGC?GATGGATTAA?GAGCTTGCTC?TTATGAAGTT?AGCGGCGGAC
GGGTGAGTAA?CACGTGGGTA?ACCTGCCCAT?AAGACTGGGA?TAACTCCGGG
AAACCGGGGC?TAATACCGGA?TAACATTTTG?AACTGCATGG?TTCGAAATTG
AAAGGCGGCT?TCGGCTGCCA?CTTATGGATG?GACCCGCGTC?GCATTAGCTA
GTTGGTGAGG?TAACGGCTCA?CCAAGGCAAC?GATGCGTAGC?CGACCTGAGA
GGGTGATCGG?CCACACTGGG?ACTGAGACAC?GGCCCAGACT?CCTACGGGAG
GCAGCAGTAG?GGAATCTTCC?GCAATGGACG?AAAGTCTGAC?GGAGCAACGC
CGCGTGAGTG?ATGAAGGCTT?TCGGGTCGTA?AAACTCTGTT?GTTAGGGAAG
AACAAGTGCT?AGTTGAATAA?GCTGGCACCT?TGACGGTACC?TAACCAGAAA
GCCACGGCTA?ACTACGTGCC?AGCAGCCGCG?GTAATACGTA?GGTGGCAAGC
GTTATCCGGA?ATTATTGGGC?GTAAAGCGCG?CGCAGGTGGT?TTCTTAAGTC
TGATGTGAAA?GCCCACGGCT?CAACCGTGGA?GGGTCATTGG?AAACTGGGAG
ACTTGAGTGC?AGAAGAGGAA?AGTGGAATTC?CATGTGTAGC?GGTGAAATGC
GTAGAGATAT?GGAGGAACAC?CAGTGGCGAA?GGCGACTTTC?TGGTCTGTAA
CTGACACTGA?GGCGCGAAAG?CGTGGGGAGC?AAACAGGATT?AGATACCCTG
GTAGTCCACG?CCGTAAACGA?TGAGTGCTAA?GTGTTAGAGG?GTTTCCGCCC
TTTAGTGCTG?AAGTTAACGC?ATTAAGCACT?CCGCCTGGGG?AGTACGGCCG
CAAGGCTGAA?ACTCAAAGGA?ATTGACGGGG?GCCCGCACAA?GCGGTGGAGC
ATGTGGTTTA?ATTCGAAGCA?ACGCGAAGAA?CCTTACCAGG?TCTTGACATC
CTCTGAAAAC?CCTAGAGATA?GGGCTTCTCC?TTCGGGAGCA?GAGTGACAGG
TGGTGCATGG?TTGTCGTCAG?CTCGTGTCGT?GAGATGTTGG?GTTAAGTCCC
GCAACGAGCG?CAACCCTTGA?TCTTAGTTGC?CATCATTAAG?TTGGGCACTC
TAAGGTGACT?GCCGGTGACA?AACCGGAGGA?AGGTGGGGAT?GACGTCAAAT
CATCATGCCC?CTTATGACCT?GGGCTACACA?CGTGCTACAA?TGGACGGTAC
AAAGAGCTGC?AAGACCGCGA?GGTGGAGCTA?ATCTCATAAA?ACCGTTCTCA
GTTCGGATTG?TAGGCTGCAA?CTCGCCTACA?TGAAGCTGGA?ATCGCTAGTA
ATCGCGGATC?AGCATGCCGC?GGTGAATACG?TTCCCGGGCC?TTGTACACAC
CGCCCGTCAC?ACCACGAGAG?TTTGTAACAC?CCGAAGTCGG?TGGGGTAACC
TTTTCAAAAT
3) gyrB gene sequencing result
TGACGGCGGC?GGTTATAAAG?TTTCTGGTGG?TTTGCATGGT?GTTGGGGCAT
CTGTAGTAAA?TGCTCTATCA?ACAGAACTAG?AGGTATTTGT?ACATCGTGAA
GGTAAAATCC?ATTATCAAAA?ATACGAAAGA?GGTATTCCAG?TTGCGGATTT
AAAAGTCATT?GGTGACACAG?ATCAAACAGG?AACGATAACT?CGATTTAAAC
CAGATCCAGA?AATTTTTCAG?GAAACAACAG?TATACGAATT?CGATACGCTA
GCAACTCGTA?TGCGTGAATT?AGCATTTTAA?ATCGTAATAT?TAAATTGACG
ATTGAAGATA?AACGTGAACA?TAAGCAAAAG?AAAGAATTCC?ATTACGAAGG
TGGAATTAAA?TCATATGTTG?AGCATTTAAA?CCGCTCAAAA?CAACCAATCC
ATGAAGAACC?TGTATATGTA?GAAGGATCAA?AAGATGGTAT?TCAAGTTGAG
GTTTCCTTAC?AGTATAACGA?AGGATATACA?AATAATATTT?ACTCATTTAC
GAATAACATT?CATACGTATG?AAGGTGGAAC?ACATGAAGTA?GGTTTTAAAA
CAGCTTTAAC?TCGTGTGATT?AACGATTACG?GTCGTAAAAA?TAGTATTTTA
AAAGATGCAG?ACAGTAACTT?AACTGGCGAG?GATGTTCGTG?AAGGTTTAAC
TGCAATCGTA?TCAATTAAAC?ATCCAAATCC?ACAATTTGAA?GGACAAACGA
AGACGAAACT?TGGGAATAGT?GAAGCGAGAA?CGATTACAGA?GTCTGTGTTT
TCAGAGGCAT?TTGAAAAGTT?CTTACTAGAA?AACCCAAACG?TTGCACGAAA
AATTGTGGAA?AAAGGTACGA?TGGCAGCACG?TGCGCGTGTT?GCAGCGAAAA
AAGCACGTGA?ATTAACACGC?CGTAAGAGTG?CGTTAGAAGT?TTCAAGCTTA
CCTGGTAAAT?TAGCAGATTG?CTCTTCAAAA?GATCCAGCAA?TTAGCGAAAT
TTATATTGTA?GAGGGTGATT?CTGCCGGCGG?ATCAGCAAAG?CAAGGTCGTG
ACCGTCACTT?CCAGGCGATT?TTACCGCTAA?AGGGTAAAAT?TATTAACGTT
GAAAAAGCAA?GATTAGATAA?AATTTTATCT?AACGATGAAG?TGCGTACAAT
TATTACTGCA?ATTGGTACGA?ACATTGGCGG?AGATTTTGAT?ATTGAGAAAG
CTCGTTATCA?TAAAGTTATT?ATTATGACGG?ACGCCGACGT?CGACGGCTCG
CACATCCG
4) fermentation condition
Seed culture medium is the NA liquid nutrient medium, and the initial pH that ferments is that 7.5,250mL triangular flask bottling amount is 150mL, 32 ℃ of shaking table revolutions are 150r/min, fermentation time is 24h, with the inoculum size of massfraction 10% be inoculated into cultivate 72h in wheat bran after, naturally dry to moisture content less than 30%.
2, Ty-3 is aspergillus niger (Aspergillus niger)
1) colonial morphology and microscopic features
The upper colony growth of wort agar substratum (MEA) is fast, lower 7 days colony diameter 50～63mm of 25 ℃ of dark conditions, and quality is velvet-like; Conidial fructification forms in a large number, and the conidial head Vandyke brown is to chocolate, and the initial stage is spherical, and the later stage splits and is several columnar structures; The bacterium colony back side is light brown, without water-soluble pigment.
Conidiophore is tall and big, wide 6～15 μ m, and wall is smooth, and is straight or crooked; The top capsule is spherical, diameter 22～30 μ m, and all surfaces can be educated; Conidial fructification is double-deck; Conidium is subsphaeroidal, and is shallow to brown, the coarse or tool spinule of wall, 3.0～4.8 μ m.There are no spermatium as shown in Figure 4.
2) rRNA gene order sequencing result:
(comprising 18S rRNA fragment, ITS1,5.8S rRNA, ITS2 district's complete sequence and 28S rRNA sequence fragment)
GATCCGAGGTCACCTGGAAAGAATGGTTGGAAAACGTCGGCAGGCGCCGGCCAATCCTACAGAGCATGTGACAAAGCCCCATACGCTCGAGGATCGGACGCGGTGCCGCCGCTGCCTTTCGGGCCCGTCCCCCCGGAGAGGGGGACGGCGACCCAACACACAAGCCGGGCTTGAGGGCAGCAATGACGCTCGGACAGGCATGCCCCCCGGAATACCAGGGGGCGCAATGTGCGTTCAAAGACTCGATGATTCACTGAATTCTGCAATTCACATTAGTTATCGCATTTCGCTGCGTTCTTCATCGATGCCGGAACCAAGAGATCCATTGTTGAAAGTTTTAACTGATTGCATTCAATCAACTCAGACTGCACGCTTTCAGACAGTGTTCGTGTTGGGGTCTCCGGCGGGCACGGGCCCGGGGGGCAGAGGCGCCCCCCCGGCGGCCCACAAGCGGCGGGCCCGCCGAAGCAACAGGGTACAATAGACACGGATGGGAGGTTGGGCCCAAAGGACCCGCACTCGGTAATGATCCTTCCGCAGGTTCACCTACGGAAACCTTGTTAC
4) fermentation condition
Seed culture medium is the NA liquid nutrient medium, and the initial pH that ferments is that 7.5,250mL triangular flask bottling amount is 150mL, 32 ℃ of shaking table revolutions are 150r/min, fermentation time is 24h, with the inoculum size of massfraction 10% be inoculated into cultivate 72h in wheat bran after, naturally dry to moisture content less than 30%.
The production of embodiment 4, Antagonistic Fungi microbial inoculum
Antagonistic Fungi QJ-1 and the Ty-3 of purifying are inoculated into respectively on the NA slant medium, cultivate 24h in the incubator of 32 ℃, get 5mL sterilized water contra bevel, vibration gently, then accurately drawing the lmL bacteria suspension joins in the 250mL triangular flask that the aseptic NA liquid nutrient medium of 150mL is housed, 1d is cultivated in the shaking bath concussion that is put into 32 ℃, then the inoculum size of cultured bacteria suspension according to massfraction 10% joined respectively in the wheat bran of sterilization, cultivate 3d, the air-dry single bacteria agent that is QJ-1 and Ty-3 for 32 ℃.In single bacteria agent, QJ-1 and Ty-3 reach respectively 10
8Then cfu/g mixes two kinds of single bacteria agents in the ratio of quality 1:1, be the Antagonistic Fungi microbial inoculum.
The production of embodiment 5, microbial organic fertilizer
By the inoculum size of massfraction 10%, the Antagonistic Fungi microbial inoculum is added in decomposed manure and mix equably, regulate moisture to 50-60%, pile the stacking of 80cm, after the 5d that banks up and get final product.
The preparation method of decomposed manure: feces of livestock and poultry is regulated moisture to 50-60%, piles 1.1m high, turning every day, and compost temperature raises gradually, and when temperature dropped near room temperature, fermentation ends was the fertilizer that becomes thoroughly decomposed.
Experimental example 1, the effect analysis of control tobacco bacterial wilt
1, carried out field experiment in Shaowu City, Fujian Province cigarette district in 2012
1) fertilizing time:
The present invention reaches the purpose that suppresses bacterial wilt by the administered twice microbial organic fertilizer.For the first time, microbial organic fertilizer is applied with the base manure form, probiotics in soil just can be rolled up in the early stage of cigarette strain growth, thereby the growth of inhibition pathogenic bacteria in soil greatly reduces its quantity.For the second time, (high density bacterial manure: the form of water=1:10) applies, and utilizes the amount reproduction of probiotics to resist the Growth and reproduction of ralstonia solanacearum to topdress when local bacterial wilt is about to occur.
2) for examination soil and block design:
Soil is sand loam, and physical features is smooth, and irrigation and drainage are convenient, and basic fertility is: organic 40.6g/kg, alkali-hydrolyzable nitrogen 198.5mg/kg, rapid available phosphorus (P
2O
5) 20.52mg/kg, available potassium (K
2O) 80.43mg/kg, pH are 5.52.Be K326 for the examination flue-cured tobacco cultivars.Adopt the floating seedlings technology to grow seedlings, transplant with seedling transplantation technique under film, manage by the requirement of High Quality Tobacco production specifications after transplanting.Each processes three repetitions, and the cultivation technique measure of each residential quarter, bookkeeping require substantially to be consistent.
Test arranges four processing altogether:
Process A: contrast, local conventional fertilizer application and management;
Treatments B: microbial organic fertilizer applies with base manure (250g/ strain) form;
Process C: microbial organic fertilizer applies with base manure (200g/ strain)+(the filling with root 50g/ strain) form of topdressing;
Process D: microbial organic fertilizer applies with (the filling with root 50g/ strain) form of topdressing.
3) researching determining project
Between local bacterial wilt period of disease, date, sickness rate and the onset speed of just morbidity of observed and recorded different treatment bacterial wilt also calculates disease index, and from morbidity, record once weekly; The severity Scaling standard is undertaken by national tobacco industry tobacco diseases investigation grade scale Yc/T39-1996, and the bacterial wilt investigation method: 0 grade, complete stool is anosis; 1 grade, the even an IOU issued by a post office spot that moves back of stem, or (with) slightly wilting with lower blade or top, or scab appears in bottom minority blade; 2 grades, there is obvious black streak in stem but does not reach cigarette strain top, or (with) sick side is more than half or part waist leaf is wilting with blade; 3 grades, stem black streak arrives cigarette strain top, or (with) sick lateral lobe sheet is all wilting, or the most of blade of complete stool is wilting; 4 grades, diseased plant is withered.Disease index and prevention effect are pressed tobacco diseases test of pesticide effectiveness method Yc/T40-1996 and are calculated:
Shaowu City, Fujian Province cigarette district field experiment result is as shown in table 1.As can be seen from Table 1, use microbial organic fertilizer of the present invention, the sickness rate of tobacco bacterial wilt has reduced more than 98%, and disease index obviously descends, and illustrates that microbial organic fertilizer of the present invention has the effect of good control tobacco bacterial wilt.
Table 1 Shaowu, Fujian Province field experiment in 2012 is prevented and treated the bacterial wilt effect
| Process | A | B | C | D |
| Sickness rate | 71.95% | 3.11% | 1.21% | 4.34% |
| Disease index | 33.83% | 0.98% | 0.67% | 1.34% |
The yield of tobacco of experimental example 2, tobacco and mass analysis
The tobacco of Shaowu, Fujian Province field experiment in 2012 in experimental example 1 is carried out yield of tobacco and mass analysis, result such as table 2.
Tobacco smoke leaf-making quantity and the mass analysis of table 2 Shaowu, Fujian Province field experiment in 2012
As can be seen from Table 2, after using microbial organic fertilizer of the present invention, the Quality and yield of tobacco tobacco leaf all is greatly improved.
Claims (4)
1. control tobacco bacterial wilt Antagonistic Fungi, is characterized in that, described Antagonistic Fungi is bacillus cereus (Bacillus cereus) QJ-1 and aspergillus niger (Aspergillus niger) Ty-3; Wherein, bacillus cereus (Bacillus cereus) QJ-1, depositary institution: Chinese Typical Representative culture collection center, preservation address: Wuhan, China Wuhan University, preservation date: on July 5th, 2012, preserving number: CCTCCNO:M2012271.
2. control tobacco bacterial wilt Antagonistic Fungi according to claim 1, it is characterized in that, described aspergillus niger (Aspergillus niger) Ty-3, depositary institution: Chinese Typical Representative culture collection center, preservation address: Wuhan, China Wuhan University, preservation date: on July 8th, 2011, preserving number: CCTCC NO:M2011241.
3. the application of control tobacco bacterial wilt Antagonistic Fungi aspect microbiobacterial agent.
4. the application of control tobacco bacterial wilt Antagonistic Fungi aspect microbial organic fertilizer.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201310046171.3A CN103146600B (en) | 2013-02-05 | 2013-02-05 | Antagonistic bacteria for prevention and treatment of tobacco bacterial wilt and application thereof |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201310046171.3A CN103146600B (en) | 2013-02-05 | 2013-02-05 | Antagonistic bacteria for prevention and treatment of tobacco bacterial wilt and application thereof |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN103146600A true CN103146600A (en) | 2013-06-12 |
| CN103146600B CN103146600B (en) | 2014-07-30 |
Family
ID=48544923
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201310046171.3A Expired - Fee Related CN103146600B (en) | 2013-02-05 | 2013-02-05 | Antagonistic bacteria for prevention and treatment of tobacco bacterial wilt and application thereof |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN103146600B (en) |
Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103865805A (en) * | 2014-01-28 | 2014-06-18 | 李祝 | Application of fermented Aspergillus niger xj for inhibiting Ralstonia solannacearum of tobacco |
| CN104450560A (en) * | 2014-11-11 | 2015-03-25 | 云南大学 | Nematode-killing sphingosine bacillus strain and application thereof |
| CN105175162A (en) * | 2015-10-22 | 2015-12-23 | 仲恺农业工程学院 | Microbial organic fertilizer for preventing bacterial wilt as well as preparation method and application thereof |
| CN106190890A (en) * | 2016-07-08 | 2016-12-07 | 郑州大学 | A kind of complex microbial inoculum preventing and treating black shank and biological organic fertilizer |
| CN114854627A (en) * | 2022-04-29 | 2022-08-05 | 重庆西农植物保护科技开发有限公司 | Pseudomonas fluorescens for preventing and treating bacterial wilt and application thereof |
Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101659932A (en) * | 2009-09-18 | 2010-03-03 | 南京农业大学 | Antagonistic bacteria preventing and removing continuous cropping tobacco bacterial wilt and microbial organic fertilizer thereof |
| CN101886055A (en) * | 2009-09-18 | 2010-11-17 | 南京农业大学 | Antagonistic bacteria NJL-14 for preventing and controlling continuous-cropping tobacco bacterial wilt |
| CN102732430A (en) * | 2011-09-05 | 2012-10-17 | 郑州大学 | Aspergillus niger strain and application thereof |
-
2013
- 2013-02-05 CN CN201310046171.3A patent/CN103146600B/en not_active Expired - Fee Related
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101659932A (en) * | 2009-09-18 | 2010-03-03 | 南京农业大学 | Antagonistic bacteria preventing and removing continuous cropping tobacco bacterial wilt and microbial organic fertilizer thereof |
| CN101886055A (en) * | 2009-09-18 | 2010-11-17 | 南京农业大学 | Antagonistic bacteria NJL-14 for preventing and controlling continuous-cropping tobacco bacterial wilt |
| CN102732430A (en) * | 2011-09-05 | 2012-10-17 | 郑州大学 | Aspergillus niger strain and application thereof |
Non-Patent Citations (1)
| Title |
|---|
| 张竹青等: "七株抗青枯病菌生防菌的初步鉴定", 《湖南农业大学学报(自然科学版)》 * |
Cited By (10)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103865805A (en) * | 2014-01-28 | 2014-06-18 | 李祝 | Application of fermented Aspergillus niger xj for inhibiting Ralstonia solannacearum of tobacco |
| CN103865805B (en) * | 2014-01-28 | 2016-05-04 | 李祝 | Fermented black aspergillus suppresses the purposes of tobacco ralstonia solanacearum |
| CN104450560A (en) * | 2014-11-11 | 2015-03-25 | 云南大学 | Nematode-killing sphingosine bacillus strain and application thereof |
| CN104450560B (en) * | 2014-11-11 | 2017-03-01 | 云南大学 | One plant of nematicide Sphingobacterium bacterial strain and its application |
| CN105175162A (en) * | 2015-10-22 | 2015-12-23 | 仲恺农业工程学院 | Microbial organic fertilizer for preventing bacterial wilt as well as preparation method and application thereof |
| CN105175162B (en) * | 2015-10-22 | 2019-01-11 | 仲恺农业工程学院 | It is a kind of for preventing and treating the microbial organic fertilizer and its preparation and application of bacterial wilt |
| CN106190890A (en) * | 2016-07-08 | 2016-12-07 | 郑州大学 | A kind of complex microbial inoculum preventing and treating black shank and biological organic fertilizer |
| CN106190890B (en) * | 2016-07-08 | 2019-05-14 | 郑州大学 | A kind of complex microbial inoculum and biological organic fertilizer for preventing and treating tobacco black shank |
| CN114854627A (en) * | 2022-04-29 | 2022-08-05 | 重庆西农植物保护科技开发有限公司 | Pseudomonas fluorescens for preventing and treating bacterial wilt and application thereof |
| CN114854627B (en) * | 2022-04-29 | 2023-10-13 | 重庆西农植物保护科技开发有限公司 | Pseudomonas fluorescens for preventing and treating bacterial wilt and application thereof |
Also Published As
| Publication number | Publication date |
|---|---|
| CN103146600B (en) | 2014-07-30 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN104531533B (en) | A kind of composite bacteria agent for preventing and treating plant continuous cropping disease and preparation method thereof, application | |
| CN103451135B (en) | Bacillus subtilis M3 and application thereof | |
| CN105886428A (en) | Streptomyces albidoflavus and applications thereof in microbial fertilizers | |
| CN106497831B (en) | A kind of prevention and treatment Phytophthora nicotianae disease composite bacteria agent and its preparation method and application | |
| CN106119155B (en) | For preventing and treating the bacterial strain WXX-2 and microbial inoculum of peanut root rot | |
| CN106588440A (en) | Special biological organic fertilizer for cucumis sativus linn succession cropping and preparation method of biological organic fertilizer | |
| CN102851219B (en) | Paecilomyces lilacinus and application thereof | |
| CN105543132A (en) | Bacillus methylotrophicus YB-F7 and application thereof in preventing plant diseases | |
| CN111690578B (en) | Salt and alkali resistant Siamese bacillus and production method and application of viable bacteria preparation thereof | |
| CN103146600B (en) | Antagonistic bacteria for prevention and treatment of tobacco bacterial wilt and application thereof | |
| CN108034601A (en) | One bacillus amyloliquefaciens and its application | |
| CN103087960A (en) | Bacillus amyloliquefaciens FQS38 and application thereof | |
| CN105132296B (en) | A kind of hook-shaped trichoderma strain and its application | |
| CN104738093A (en) | Preparation method for bacillus coagulans bacterial suspension | |
| CN106119154A (en) | A kind of bacterial strain preventing and treating Peanut continuous cropping silborne fungal diseases and application | |
| CN106222121B (en) | A kind of bacillus megaterium bacterial strain, biocontrol agent and the preparation method and application thereof | |
| CN105154339A (en) | Trichoderma viride strain and application thereof | |
| CN102925387A (en) | Bacillus simplex for inducing soybean to generate soybean cyst nematode resistance and application | |
| CN110122485A (en) | A kind of trichoderma, Trichoderma and preparation method thereof | |
| CN110317747A (en) | A kind of bacillus amyloliquefaciens JT68 and its application in prevention and treatment tea anthracnose | |
| CN109749953A (en) | Bacillus cereus, microbial inoculum and its preparation method and application | |
| CN105439657A (en) | Preparation method for strawberry dedicated anti-continuous cropping biological organic fertilizer | |
| CN102443559B (en) | Baclillus subtilis used for controlling cotton verticillium wilt and application thereof | |
| CN102732430A (en) | Aspergillus niger strain and application thereof | |
| CN107502584A (en) | One fluorescent pseudomonads and its application in walnut black spot is prevented and treated |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| ASS | Succession or assignment of patent right |
Owner name: ZHENGZHOU UNIVERSITY Effective date: 20131101 Owner name: SHAOWU SUBBRANCH OF NANPING TOBACCO CORPORATION Free format text: FORMER OWNER: ZHENGZHOU UNIVERSITY Effective date: 20131101 |
|
| C41 | Transfer of patent application or patent right or utility model | ||
| C53 | Correction of patent of invention or patent application | ||
| CB03 | Change of inventor or designer information |
Inventor after: Xie Tingxin Inventor after: Li Hongli Inventor after: Zeng Qiang Inventor after: Li Xiaolong Inventor after: Lu Aqian Inventor after: Wang Yan Inventor before: Li Hongli Inventor before: Lu Aqian Inventor before: Wang Yan |
|
| COR | Change of bibliographic data |
Free format text: CORRECT: INVENTOR; FROM: LI HONGLI LU AQIAN WANG YAN TO: XIE TINGXIN LI HONGLI CENG QIANG LI XIAOLONG LU AQIAN WANG YAN Free format text: CORRECT: ADDRESS; FROM: 450001 ZHENGZHOU, HENAN PROVINCE TO: 354000 NANPING, FUJIAN PROVINCE |
|
| TA01 | Transfer of patent application right |
Effective date of registration: 20131101 Address after: 354000 Shaowu province Fujian City Huaguang Road Garden Office Minxing tobacco Applicant after: Shaowu Branch, Nanping Tobacco Co.,Ltd. Applicant after: Zhengzhou University Address before: 450001 science Avenue, Henan, Zhengzhou, No. 100 Applicant before: Zhengzhou University |
|
| C14 | Grant of patent or utility model | ||
| GR01 | Patent grant | ||
| CF01 | Termination of patent right due to non-payment of annual fee | ||
| CF01 | Termination of patent right due to non-payment of annual fee |
Granted publication date: 20140730 Termination date: 20180205 |