CN100370018C - Segmented oxygen supply fermentation process of entomopathogenic nematode symbiotic bacteria and use of fermentation product thereof - Google Patents
Segmented oxygen supply fermentation process of entomopathogenic nematode symbiotic bacteria and use of fermentation product thereof Download PDFInfo
- Publication number
- CN100370018C CN100370018C CNB2005100428227A CN200510042822A CN100370018C CN 100370018 C CN100370018 C CN 100370018C CN B2005100428227 A CNB2005100428227 A CN B2005100428227A CN 200510042822 A CN200510042822 A CN 200510042822A CN 100370018 C CN100370018 C CN 100370018C
- Authority
- CN
- China
- Prior art keywords
- substratum
- oxygen supply
- xenorhabdus nematophilus
- fermentation
- culture medium
- 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.)
- Expired - Fee Related
Links
- 241000894006 Bacteria Species 0.000 title claims abstract description 40
- 241000244206 Nematoda Species 0.000 title claims abstract description 31
- QVGXLLKOCUKJST-UHFFFAOYSA-N atomic oxygen Chemical compound [O] QVGXLLKOCUKJST-UHFFFAOYSA-N 0.000 title claims abstract description 22
- 239000001301 oxygen Substances 0.000 title claims abstract description 22
- 229910052760 oxygen Inorganic materials 0.000 title claims abstract description 22
- 238000000855 fermentation Methods 0.000 title claims abstract description 17
- 230000004151 fermentation Effects 0.000 title claims abstract description 17
- 230000000967 entomopathogenic effect Effects 0.000 title claims description 21
- 241000607735 Xenorhabdus nematophila Species 0.000 claims abstract description 23
- 238000000034 method Methods 0.000 claims abstract description 14
- 201000010099 disease Diseases 0.000 claims abstract description 13
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 claims abstract description 13
- 230000008569 process Effects 0.000 claims abstract description 10
- 239000000463 material Substances 0.000 claims abstract description 7
- 239000001963 growth medium Substances 0.000 claims description 24
- 239000007788 liquid Substances 0.000 claims description 23
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 23
- 239000002609 medium Substances 0.000 claims description 18
- 238000002156 mixing Methods 0.000 claims description 17
- 239000001888 Peptone Substances 0.000 claims description 13
- 108010080698 Peptones Proteins 0.000 claims description 13
- 235000019319 peptone Nutrition 0.000 claims description 13
- PEDCQBHIVMGVHV-UHFFFAOYSA-N Glycerine Chemical compound OCC(O)CO PEDCQBHIVMGVHV-UHFFFAOYSA-N 0.000 claims description 11
- 238000011218 seed culture Methods 0.000 claims description 8
- 239000011734 sodium Substances 0.000 claims description 8
- 230000001954 sterilising effect Effects 0.000 claims description 7
- 229940041514 candida albicans extract Drugs 0.000 claims description 6
- 238000005516 engineering process Methods 0.000 claims description 6
- 239000006916 nutrient agar Substances 0.000 claims description 6
- 239000012138 yeast extract Substances 0.000 claims description 6
- 239000012752 auxiliary agent Substances 0.000 claims description 5
- 230000001276 controlling effect Effects 0.000 claims description 5
- 239000013530 defoamer Substances 0.000 claims description 5
- 235000011187 glycerol Nutrition 0.000 claims description 5
- 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 4
- 239000013543 active substance Substances 0.000 claims description 4
- WPYMKLBDIGXBTP-UHFFFAOYSA-N benzoic acid Chemical compound OC(=O)C1=CC=CC=C1 WPYMKLBDIGXBTP-UHFFFAOYSA-N 0.000 claims description 4
- 239000008103 glucose Substances 0.000 claims description 4
- 239000002054 inoculum Substances 0.000 claims description 4
- 230000001105 regulatory effect Effects 0.000 claims description 4
- WXMKPNITSTVMEF-UHFFFAOYSA-M sodium benzoate Chemical compound [Na+].[O-]C(=O)C1=CC=CC=C1 WXMKPNITSTVMEF-UHFFFAOYSA-M 0.000 claims description 4
- 239000004299 sodium benzoate Substances 0.000 claims description 4
- 235000010234 sodium benzoate Nutrition 0.000 claims description 4
- 238000000151 deposition Methods 0.000 claims description 3
- BEFDCLMNVWHSGT-UHFFFAOYSA-N ethenylcyclopentane Chemical compound C=CC1CCCC1 BEFDCLMNVWHSGT-UHFFFAOYSA-N 0.000 claims description 3
- 239000004334 sorbic acid Substances 0.000 claims description 3
- 235000010199 sorbic acid Nutrition 0.000 claims description 3
- 229940075582 sorbic acid Drugs 0.000 claims description 3
- 239000000203 mixture Substances 0.000 claims description 2
- 239000002994 raw material Substances 0.000 claims description 2
- RTZKZFJDLAIYFH-UHFFFAOYSA-N Diethyl ether Chemical compound CCOCC RTZKZFJDLAIYFH-UHFFFAOYSA-N 0.000 claims 2
- 229920002503 polyoxyethylene-polyoxypropylene Polymers 0.000 claims 1
- 230000000844 anti-bacterial effect Effects 0.000 abstract description 17
- 238000003756 stirring Methods 0.000 abstract description 6
- 241000238631 Hexapoda Species 0.000 abstract description 4
- 230000010261 cell growth Effects 0.000 abstract description 3
- 239000003899 bactericide agent Substances 0.000 abstract description 2
- 230000001717 pathogenic effect Effects 0.000 abstract description 2
- 238000009423 ventilation Methods 0.000 abstract 1
- 239000000047 product Substances 0.000 description 24
- 230000002538 fungal effect Effects 0.000 description 19
- 239000000126 substance Substances 0.000 description 14
- 239000000243 solution Substances 0.000 description 12
- 241000233616 Phytophthora capsici Species 0.000 description 9
- 241000607757 Xenorhabdus Species 0.000 description 8
- 241000196324 Embryophyta Species 0.000 description 6
- 235000007688 Lycopersicon esculentum Nutrition 0.000 description 6
- 241001148062 Photorhabdus Species 0.000 description 6
- 240000003768 Solanum lycopersicum Species 0.000 description 6
- 230000000845 anti-microbial effect Effects 0.000 description 6
- 230000000452 restraining effect Effects 0.000 description 6
- 240000008067 Cucumis sativus Species 0.000 description 5
- 235000010799 Cucumis sativus var sativus Nutrition 0.000 description 5
- 239000007864 aqueous solution Substances 0.000 description 5
- 239000003814 drug Substances 0.000 description 5
- 230000012010 growth Effects 0.000 description 5
- 239000006228 supernatant Substances 0.000 description 5
- 235000019354 vermiculite Nutrition 0.000 description 5
- 229910052902 vermiculite Inorganic materials 0.000 description 5
- 239000010455 vermiculite Substances 0.000 description 5
- 235000002566 Capsicum Nutrition 0.000 description 4
- WSFSSNUMVMOOMR-UHFFFAOYSA-N Formaldehyde Chemical compound O=C WSFSSNUMVMOOMR-UHFFFAOYSA-N 0.000 description 4
- -1 LB substratum Substances 0.000 description 4
- 239000006002 Pepper Substances 0.000 description 4
- 241000722363 Piper Species 0.000 description 4
- 235000016761 Piper aduncum Nutrition 0.000 description 4
- 235000017804 Piper guineense Nutrition 0.000 description 4
- 235000008184 Piper nigrum Nutrition 0.000 description 4
- 241001480238 Steinernema Species 0.000 description 4
- 230000001580 bacterial effect Effects 0.000 description 4
- 235000013399 edible fruits Nutrition 0.000 description 4
- 238000011534 incubation Methods 0.000 description 4
- 238000004519 manufacturing process Methods 0.000 description 4
- 230000003449 preventive effect Effects 0.000 description 4
- 229930000044 secondary metabolite Natural products 0.000 description 4
- 238000012360 testing method Methods 0.000 description 4
- 241000588921 Enterobacteriaceae Species 0.000 description 3
- QIVBCDIJIAJPQS-VIFPVBQESA-N L-tryptophane Chemical compound C1=CC=C2C(C[C@H](N)C(O)=O)=CNC2=C1 QIVBCDIJIAJPQS-VIFPVBQESA-N 0.000 description 3
- QIVBCDIJIAJPQS-UHFFFAOYSA-N Tryptophan Natural products C1=CC=C2C(CC(N)C(O)=O)=CNC2=C1 QIVBCDIJIAJPQS-UHFFFAOYSA-N 0.000 description 3
- 239000006035 Tryptophane Substances 0.000 description 3
- 241000123579 Xenorhabdus bovienii Species 0.000 description 3
- 238000002791 soaking Methods 0.000 description 3
- 229960004799 tryptophan Drugs 0.000 description 3
- 238000005406 washing Methods 0.000 description 3
- NIQUZFQMNMNAMD-NSHDSACASA-N (3s)-n-[2-(1h-indol-3-yl)ethyl]-3-methyl-2-oxopentanamide Chemical compound C1=CC=C2C(CCNC(=O)C(=O)[C@@H](C)CC)=CNC2=C1 NIQUZFQMNMNAMD-NSHDSACASA-N 0.000 description 2
- 235000000832 Ayote Nutrition 0.000 description 2
- 240000004244 Cucurbita moschata Species 0.000 description 2
- 235000009854 Cucurbita moschata Nutrition 0.000 description 2
- 235000009804 Cucurbita pepo subsp pepo Nutrition 0.000 description 2
- 241000233866 Fungi Species 0.000 description 2
- 241000223221 Fusarium oxysporum Species 0.000 description 2
- 240000003978 Ipomoea coccinea Species 0.000 description 2
- 241001330975 Magnaporthe oryzae Species 0.000 description 2
- 244000061176 Nicotiana tabacum Species 0.000 description 2
- 235000002637 Nicotiana tabacum Nutrition 0.000 description 2
- 240000007594 Oryza sativa Species 0.000 description 2
- 235000007164 Oryza sativa Nutrition 0.000 description 2
- 241000191967 Staphylococcus aureus Species 0.000 description 2
- 230000000259 anti-tumor effect Effects 0.000 description 2
- 230000003385 bacteriostatic effect Effects 0.000 description 2
- 230000004071 biological effect Effects 0.000 description 2
- 229910052799 carbon Inorganic materials 0.000 description 2
- 239000000645 desinfectant Substances 0.000 description 2
- 230000000694 effects Effects 0.000 description 2
- 230000035558 fertility Effects 0.000 description 2
- 244000000004 fungal plant pathogen Species 0.000 description 2
- 150000002475 indoles Chemical class 0.000 description 2
- 238000002955 isolation Methods 0.000 description 2
- 239000002207 metabolite Substances 0.000 description 2
- ZQEIXNIJLIKNTD-UHFFFAOYSA-N methyl N-(2,6-dimethylphenyl)-N-(methoxyacetyl)alaninate Chemical compound COCC(=O)N(C(C)C(=O)OC)C1=C(C)C=CC=C1C ZQEIXNIJLIKNTD-UHFFFAOYSA-N 0.000 description 2
- NIQUZFQMNMNAMD-UHFFFAOYSA-N nematophin Natural products C1=CC=C2C(CCNC(=O)C(=O)C(C)CC)=CNC2=C1 NIQUZFQMNMNAMD-UHFFFAOYSA-N 0.000 description 2
- 239000000575 pesticide Substances 0.000 description 2
- 238000012545 processing Methods 0.000 description 2
- 235000015136 pumpkin Nutrition 0.000 description 2
- RXWNCPJZOCPEPQ-NVWDDTSBSA-N puromycin Chemical compound C1=CC(OC)=CC=C1C[C@H](N)C(=O)N[C@H]1[C@@H](O)[C@H](N2C3=NC=NC(=C3N=C2)N(C)C)O[C@@H]1CO RXWNCPJZOCPEPQ-NVWDDTSBSA-N 0.000 description 2
- 235000009566 rice Nutrition 0.000 description 2
- 238000012216 screening Methods 0.000 description 2
- 239000002689 soil Substances 0.000 description 2
- 238000004659 sterilization and disinfection Methods 0.000 description 2
- 229960005486 vaccine Drugs 0.000 description 2
- 241000193755 Bacillus cereus Species 0.000 description 1
- 241000123650 Botrytis cinerea Species 0.000 description 1
- OKTJSMMVPCPJKN-UHFFFAOYSA-N Carbon Chemical compound [C] OKTJSMMVPCPJKN-UHFFFAOYSA-N 0.000 description 1
- 235000016623 Fragaria vesca Nutrition 0.000 description 1
- 240000009088 Fragaria x ananassa Species 0.000 description 1
- 235000011363 Fragaria x ananassa Nutrition 0.000 description 1
- 241001523406 Heterorhabditis Species 0.000 description 1
- 241000500097 Heterorhabditis indica Species 0.000 description 1
- SAQNYTQFLPVTNJ-UHFFFAOYSA-N Madumycin II Natural products C1C(O)CC(O)C=C(C)C=CCNC(=O)C=CC(C)C(C(C)C)OC(=O)C(C)NC(=O)C2=COC1=N2 SAQNYTQFLPVTNJ-UHFFFAOYSA-N 0.000 description 1
- 241000589516 Pseudomonas Species 0.000 description 1
- 240000004808 Saccharomyces cerevisiae Species 0.000 description 1
- 241000607142 Salmonella Species 0.000 description 1
- 241000187747 Streptomyces Species 0.000 description 1
- 241001041742 Xenorhabdus miraniensis Species 0.000 description 1
- 230000003698 anagen phase Effects 0.000 description 1
- 150000004056 anthraquinones Chemical class 0.000 description 1
- 239000003242 anti bacterial agent Substances 0.000 description 1
- 230000001857 anti-mycotic effect Effects 0.000 description 1
- 230000002155 anti-virotic effect Effects 0.000 description 1
- 229940088710 antibiotic agent Drugs 0.000 description 1
- 239000002543 antimycotic Substances 0.000 description 1
- 239000003699 antiulcer agent Substances 0.000 description 1
- 244000052616 bacterial pathogen Species 0.000 description 1
- 230000015572 biosynthetic process Effects 0.000 description 1
- 210000000481 breast Anatomy 0.000 description 1
- 150000001875 compounds Chemical class 0.000 description 1
- 239000007859 condensation product Substances 0.000 description 1
- 230000007812 deficiency Effects 0.000 description 1
- 239000012895 dilution Substances 0.000 description 1
- 238000010790 dilution Methods 0.000 description 1
- 239000012153 distilled water Substances 0.000 description 1
- 230000000857 drug effect Effects 0.000 description 1
- 239000000975 dye Substances 0.000 description 1
- 238000009313 farming Methods 0.000 description 1
- 238000011010 flushing procedure Methods 0.000 description 1
- 239000008098 formaldehyde solution Substances 0.000 description 1
- 230000000855 fungicidal effect Effects 0.000 description 1
- 239000000417 fungicide Substances 0.000 description 1
- 150000002391 heterocyclic compounds Chemical class 0.000 description 1
- 238000000338 in vitro Methods 0.000 description 1
- 230000002401 inhibitory effect Effects 0.000 description 1
- 230000005764 inhibitory process Effects 0.000 description 1
- 108010053119 madumycin II Proteins 0.000 description 1
- VQNWNKSWWORSDJ-QBPIOVJMSA-N madumycin ii Chemical compound C1C(=O)CC(O)\C=C(/C)\C=C\CNC(=O)\C=C/C(C)C(C(C)C)OC(=O)C(C)NC(=O)C2=COC1=N2 VQNWNKSWWORSDJ-QBPIOVJMSA-N 0.000 description 1
- 230000003211 malignant effect Effects 0.000 description 1
- 239000000155 melt Substances 0.000 description 1
- 229960002523 mercuric chloride Drugs 0.000 description 1
- LWJROJCJINYWOX-UHFFFAOYSA-L mercury dichloride Chemical compound Cl[Hg]Cl LWJROJCJINYWOX-UHFFFAOYSA-L 0.000 description 1
- 230000002906 microbiologic effect Effects 0.000 description 1
- 239000005645 nematicide Substances 0.000 description 1
- 229920005646 polycarboxylate Polymers 0.000 description 1
- 238000002360 preparation method Methods 0.000 description 1
- 108090000623 proteins and genes Proteins 0.000 description 1
- 102000004169 proteins and genes Human genes 0.000 description 1
- 229950010131 puromycin Drugs 0.000 description 1
- 230000002829 reductive effect Effects 0.000 description 1
- 238000012552 review Methods 0.000 description 1
- 239000013535 sea water Substances 0.000 description 1
- 238000007789 sealing Methods 0.000 description 1
- SUKJFIGYRHOWBL-UHFFFAOYSA-N sodium hypochlorite Chemical compound [Na+].Cl[O-] SUKJFIGYRHOWBL-UHFFFAOYSA-N 0.000 description 1
- 229920005552 sodium lignosulfonate Polymers 0.000 description 1
- 159000000000 sodium salts Chemical class 0.000 description 1
- 238000001228 spectrum Methods 0.000 description 1
- 239000007921 spray Substances 0.000 description 1
- 238000007619 statistical method Methods 0.000 description 1
- 230000031068 symbiosis, encompassing mutualism through parasitism Effects 0.000 description 1
- 238000003786 synthesis reaction Methods 0.000 description 1
- 239000008399 tap water Substances 0.000 description 1
- 235000020679 tap water Nutrition 0.000 description 1
- 238000010998 test method Methods 0.000 description 1
- RTKIYNMVFMVABJ-UHFFFAOYSA-L thimerosal Chemical compound [Na+].CC[Hg]SC1=CC=CC=C1C([O-])=O RTKIYNMVFMVABJ-UHFFFAOYSA-L 0.000 description 1
- 239000003053 toxin Substances 0.000 description 1
- 231100000765 toxin Toxicity 0.000 description 1
- 239000003643 water by type Substances 0.000 description 1
Landscapes
- Agricultural Chemicals And Associated Chemicals (AREA)
Abstract
The invention relates to a segmented oxygen supply fermentation process of insect pathogenic nematode symbiotic bacteria antibacterial activity and application of fermentation liquor thereof. The process takes Xenorhabdus nematophilus YL001 as a material, and oxygen is supplied in a segmented manner by adjusting ventilation volume and stirring speed in the fermentation process. The fermentation process of the present invention has cell growth amount and antibacterial activity higher than that of single oxygen supply fermentation. The fermentation broth obtained by the fermentation process can be used for preparing a bactericide for preventing and treating plant diseases.
Description
Technical field
The present invention relates to the zymotechnique of a kind of bacterium, is a kind of sectional oxygen supply fermentation technology and fermented product purposes thereof that improves the entomopathogenic nematode symbiotic bacteria anti-microbial activity specifically.
Background technology
Entomopathogenic nematode symbiotic bacteria is a bacterioid that parasitizes in the entomopathogenic nematode enteron aisle, belongs to enterobacteriaceae (Enterobacteriaceae), amphimicrobian, chemoheterotrophy, Gram-negative.Comprise that Xenorhabdus belongs to (Xenorhabdus) and polished rod shape Pseudomonas (Photorhabdus), belong to the symbiosis of (Heterorhabditis) nematode with genus steinernema (Steinernema) and heterorhabditis indica respectively.At nature, this bacterioid is present in and infects for three ages in the enteron aisle of phase nematode, along with nematode infecting insect, fungal component is carried in the insect body and is discharged in host's haemocoele, and fungal component breeds in a large number, produces toxin and antibacterial substance, with the nematode acting in conjunction, kill host insect.
Have the hypothesis of antimicrobial acivity since Dutky (1959) proposes entomopathogenic nematode symbiotic bacteria, just paul et al. ability isolation identification from fungal component Xenorhabdus spp. went out several bacteriostatic compounds up to 1981.The chemical nature of the antibacterial substance that fungal component is produced and biological activity thereof are existing comparatively detailed explains (Frost, S.and Nealson, K. (1996) Molecular biology of thesymbiotic-pathogenic bacteriaXenorhabdus spp.and Photorhabdus spp.Microbiological Reviews 60,21-43.; Li, J., Hu, K.and Webster, J.M. (1998) Antibiotics from Xenorhabdus spp.and Photorhabdusspp. (Enterobacteriaceae) .Chemistry of Heterocyclic Compounds 34,1561-1570.).The antibacterial substance that fungal component produces has wider antimicrobial spectrum, inhibited to bacterium, fungi (comprising the human disease bacterium) and yeast, and some human malignant bacterias that multiple medicine produces resistance had stronger restraining effect, also has anti-tumor activity, pharmaceutically have bigger application potential, more particularly having antimycotic and meta-bolites and derivative thereof eelworm-killing activity application promise in clinical practice is arranged on agricultural.
Up to the present, from Xenorhabdus and Photorhabdus isolation identification go out the secondary metabolite of more than 30 kind of biologically active.Comprising puromycin and madumycin II, these two kinds of materials at first are isolated from Streptomyces.The kind of Xenorhabdus secondary metabolite is more than Photorhabdus.Xenorhabdus bovienii mainly produces Xenorhabdins, and Xenorhabdus nematophilus produces Xenocoumacins, and Photorhabdus then mainly produces Hydroxystibenes and Anthraquinones.These meta-bolitess not only have the number of chemical structure, and on medicine and agricultural wide biological activity are arranged, as antibacterium, fungi, desinsection, nematicide, antiulcer agent, antitumor and antivirus action.
The generation of antibacterial substance is relevant with bacterial strain of fungal component and culture condition.Xenorhabdus spp does not produce antibacterial substance in 1.0% protein culture medium, and can produce antibacterial substance in yeast extract medium, LB substratum, sea water medium and TSB substratum.Xenocoumacins1 that X.nematophilus ALL produces in the TSB substratum and 2 output are 300 and 100mg.L
-1In yeast extract medium, Xenorhabdus sp.Q1 is under the cultured continuously condition, and Xenorhabdins1 and 2 output are 10.8 and 36.4mg.L
-1In the TSB substratum, X.bovienii is the main Xenorhabdins that produces under the batch culture condition.
The generation of different incubation time antibacterial substances also has certain difference.Fungal component is in the TSB substratum, and the concentration of nematophin differs 5 times at different incubation times.Concentration at the 1-2d nematophin that cultivates sharply increases, and maintains higher level then.Indoles2 that X.bovienii produces and the output of indoles4 reach maximum at the 1d that cultivates, and are respectively 17.00 and 11.56 μ g.ml
-1, output reduces gradually then; And indoles1 and indoles3 are respectively 24.42 and 18.87 μ g.ml in the output minimum of cultivating 1d
-1, increase gradually with the prolongation output of incubation time.This explanation indoles2 and indoles4 are at the fungal component early stage synthetic of growing, and are degraded to indoles1 and indoles3 subsequently gradually.Indoles is by the tryptophane synthetic, and it has kept the mesomethylene carbon on the tryptophane, has removed carboxyl carbon, and this is because add the output that tryptophane can improve indoles in substratum.The amount of the ST that P.luminscens produces under the different culture condition has bigger difference, promptly is same bacterial strain.
Kenel variation, culture temperature and the growth of dissolved oxygen condition influence fungal component and the generation of antibacterial substance.Nascent type fungal component can produce antibacterial substance, and secondary type then can not; In vitro culture, kenel can take place and change in fungal component, and this is to influence the subject matter that ablastins produce.X.nematophilusD1 at 35 ℃ bacteriostatic activity far below 15-30 ℃.Xenorhabdus and Photorhabdus are the facultative bacterias of detesting, and oxygen is that its growth and antibacterial substance produce necessary.Fungal component does not produce ablastins under non-oscillating condition in the container of sealing.
As previously mentioned, fungal component has certain difference at different substratum with different incubation time secondary metabolites; Its kenel variation, culture temperature and dissolved oxygen condition etc. influence the growth of fungal component and the generation of antibacterial substance.The correlation technique of the generation of the cultivation and fermentation of fungal component and secondary metabolite and the document or the patent of method are not appeared in the newspapers.
Summary of the invention
The objective of the invention is at the deficiency of entomopathogenic nematode symbiotic bacteria on cultivation and fermentation technology and method, with Xenorhabdus nematophilus Xenorhabdus nematophilus YL001 is that (this bacterium is to separate to obtain from a kind of nematode Steinernema sp of Shaanxi Yang Ling screening to material, through identifying and called after Xenorhabdus nema tophilus YL001.)。Set up the zymotechnique of a set segmented oxygen supply raising entomopathogenic nematode symbiotic bacteria anti-microbial activity at this bacterium.Concrete processing step of the present invention is:
1. kind is guaranteed the Xenorhabdus nematophilus of depositing, lines on the NA culture medium flat plate that cultivate 24～48h for 28 ℃, picking list bacterium colony lines on the NBTA culture medium flat plate again, cultivates 24～48h for 28 ℃;
2. with the blue colonies on the transfering loop picking NBTA culture medium flat plate, be inoculated in the 250ml triangular flask that 50ml NB substratum is housed, 28 ℃, 180r/min shaking culture 16-24h become primary seed solution;
3. primary seed solution is equipped with in the 250ml triangular flask of seed culture medium by 9.0% inoculum size access, liquid amount is 25ml, and pH 7.2, at 26 ℃, 220rpm shaking culture 16-24h, become secondary seed solution;
4. in the 7L stirred-tank fermenter, add the 4.5L fermention medium, 0.01% defoamer, pH 7.2, and the 0.1-0.15MPa steam sterilizing 30min that offs normal is cooled to 26 ℃, inserts secondary seed solution, 26 ℃ of cultivation 72h, acquisition fermented product by 9.0%.Fermenting process is regulated air flow and mixing speed, and sectional oxygen supply can obtain fermented product, and fermented product is filtered, and removes cell and obtains fermented liquid.
Wherein said NA substratum is: extractum carnis 3.0g, peptone 5.0g, nutrient agar medium 20g, water 1000ml, PH 7.2～7.4.
Wherein said NBTA substratum is: NA+ (TTC 0.04g, BTB 0.025g).
Wherein said NB substratum is: extractum carnis 3.0g, peptone 5.0g, water 1000ml, PH 7.2～7.4.
Wherein said seed culture medium is: glycerine 5g, yeast extract 15g, 1M-MgSO
45ml, (NH
4)
2SO
42g, 1M-KH
2PO
45ml, 1M-K
2HPO
45ml, 1M-Na
2SO
410ml, water 1000ml, PH 7.2.
Wherein said fermention medium is (g/L): glucose 6.13, peptone 21.29, MgSO
41.50, (NH
4)
2SO
42.46, KH
2PO
40.86, K
2HPO
41.11, Na
2SO
41.72.
Wherein fermenting process adjusting air flow and mixing speed sectional oxygen supply pattern are: 0-10h, air flow 1.5L/min, mixing speed 200rpm; 10-40h, air flow 2.5L/min, mixing speed 200rpm → 425rpm → 250rpm; 40-72h, air flow 2.0L/min, mixing speed 250rpm.
The application of the fermented liquid of Xenorhabdus nematophilus Xenorhabdus nematophilus YL001 is to prepare disinfectant use in agriculture, and this sterilant is used for controlling plant diseases.
This sterilant is made of hundred parts of ratios of volume of following component: the fermented liquid 60%-85% of Xenorhabdus nematophilus Xenorhabdusnematophilus YL001, and auxiliary agent 1%-5%, surplus is tensio-active agent, the summation of said components is 100%.
Described auxiliary agent is a kind of in the materials such as phenylformic acid, Sodium Benzoate, Sorbic Acid, and tensio-active agent is the composition of the known material of this area staff.
Process characteristic of the present invention: adopt by regulating the new process for fermenting of air flow and mixing speed sectional oxygen supply during the fermentation, can satisfy fungal component, help the generation of cell growth and metabolite in the demand of different growth phases to oxygen.Not only improve increment, yield and the production intensity of cell, and improved output, yield and the production intensity of antimicrobial substance, thereby improved utilization ratio of raw materials, reduced production cost.
Early vaccine, pumpkin Fusarium oxysporum, cucumber anthrax-bacilus, rice blast fungus and phytophthora blight of pepper have stronger restraining effect to the fermented product of this bacterium to the red star bacterium of plant pathogenic fungi tobacco, tomato; Bacteriums such as paddy rice bacterial leaf spot bacterium, streptococcus aureus, bacillus cereus also there is stronger inhibition activity.Fermented product can be used for preparing disinfectant use in agriculture, controlling plant diseases.
Embodiment
Below content of the present invention is further elaborated by following more excellent embodiment, but these embodiment do not limit protection scope of the present invention.
The acquisition of Xenorhabdus nematophilus Xenorhabdus nematophilus YL001:
A, at first will contaminate entomopathogenic nematode (IJs) (this nematode obtains from Shaanxi Yang Ling screening, through the being accredited as Stahli line worm Steinernema sp) soaking disinfection 3 times of phase with 0.1% formaldehyde solution, each 30min uses aseptic water washing after sterilize at every turn; With 0.1% merthiolate nematode is soaked 30min subsequently, back aseptic water washing 3 times of having sterilized, the nematode that will sterilize is added on the NBTA culture medium flat plate and separates at last;
C, the clorox with 10% or 1% mercuric chloride will infect the phase entomopathogenic nematode and soak 30min, use aseptic water washing then 3 times, the nematode of sterilizing directly is dispersed on the NBTA substratum separate.The NBTA culture medium flat plate is cultivated 3-5d under 28 ℃, dark condition, nematode discharges fungal component, forms the blue-greenish colour bacterium colony, and the dyestuff in the substratum is absorbed and becomes yellow on every side, can obtain entomopathogenic nematode symbiotic bacteria, through identifying and called after Xenorhabdus nematophilus YL001.
Wherein said NBTA substratum is: extractum carnis 3.0g, peptone 5.0g, nutrient agar medium 20g, TTC 0.04g, BTB 0.025g, water 1000ml, PH 7.2～7.4.
Embodiment 1:
1. kind is guaranteed the Xenorhabdus nematophilus Xenorhabdus nematophilus YL001 that deposits, lines on the NA culture medium flat plate that cultivate 24h for 28 ℃, picking list bacterium colony lines on the NBTA culture medium flat plate again, cultivates 48h for 28 ℃;
2, with the blue colonies on the transfering loop picking NBTA culture medium flat plate, be inoculated in the 250ml triangular flask that 50ml NB substratum is housed, 28 ℃, 180r/min shaking culture 16-24h become primary seed solution;
3, primary seed solution is equipped with in the 250ml triangular flask of seed culture medium by 9.0% inoculum size access, liquid amount is 25ml, and pH7.25 at 26 ℃, 220rpm shaking culture 16-24h, becomes secondary seed solution;
4, in the 7L stirred-tank fermenter, add the 4.5L fermention medium, 0.01% defoamer, pH 7.2, the 0.1-0.15MPa steam sterilizing 30min that offs normal is cooled to 28 ℃, insert secondary seed solution by 9.0%, under 26 ℃, air flow 2.5L/min, mixing speed 300rpm condition, cultivate 72h, obtain fermented product.
The NA substratum is: extractum carnis 3.0g, peptone 5.0g, nutrient agar medium 20g, water 1000ml, PH 7.2～7.4.
The NBTA substratum is: NA+ (TTC 0.04g, BTB 0.025g).
The NB substratum is: extractum carnis 3.0g, peptone 5.0g, water 1000ml, PH 7.2～7.4.Seed culture medium is: glycerine 5g, yeast extract 15g, 1M-MgSO
45ml, (NH
4)
2SO
42g, 1M-KH
2PO
45ml, 1M-K
2HPO
45ml, 1M-Na
2SO
410ml, water 1000ml, PH 7.2.Fermention medium is (g/L) glucose 6.13, peptone 21.29, MgSO
41.50, (NH
4)
2SO
42.46, KH
2PO
40.86, K
2HPO
41.11, Na
2SO
41.72.
Embodiment 2:
1, kind is guaranteed the fungal component of depositing, lines on the NA culture medium flat plate that cultivate 24～48h for 28 ℃, picking list bacterium colony lines on the NBTA culture medium flat plate again, cultivates 24～48h for 28 ℃;
2, with the blue colonies on the transfering loop picking NBTA culture medium flat plate, be inoculated in the 250ml triangular flask that 50ml NB substratum is housed, 28 ℃, 180r/min shaking culture 16-24h become primary seed solution;
3, primary seed solution is equipped with in the 250ml triangular flask of seed culture medium by 9.0% inoculum size access, liquid amount is 25ml, and pH 7.2, at 26 ℃, 220rpm shaking culture 16-24h, become secondary seed solution;
4, in the 7L stirred-tank fermenter, add the 4.5L fermention medium, 0.01% defoamer, pH 7.2, and the 0.1-0.15MPa steam sterilizing 30min that offs normal is cooled to 26 ℃, inserts secondary seed solution, 26 ℃ of cultivation 72h, acquisition fermented product by 9.0%.Fermenting process is regulated air flow and mixing speed, sectional oxygen supply.
Wherein said NA substratum is: extractum carnis 3.0g, peptone 5.0g, nutrient agar medium 20g, water 1000ml, PH 7.2～7.4.
Wherein said NBTA substratum is: NA+ (TTC 0.04g, BTB 0.025g).
Wherein said NB substratum is: extractum carnis 3.0g, peptone 5.0g, water 1000ml, PH 7.2～7.4.
Wherein said seed culture medium is: glycerine 5g, yeast extract 15g, 1M-MgSO
45ml, (NH
4)
2SO
42g, 1M-KH
2PO
45ml, 1M-K
2HPO
45ml, 1M-Na
2SO
410ml, water 1000ml, PH 7.2.
Wherein said fermention medium is (g/L): glucose 6.13, peptone 21.29, MgSO
41.50, (NH
4)
2SO
42.46, KH
2PO
40.86, K
2HPO
41.11, Na
2SO
41.72.
Wherein fermenting process adjusting air flow and mixing speed sectional oxygen supply pattern are: 0-10h, air flow 1.5L/min, mixing speed 200rpm; 10-40h, air flow 2.5L/min, mixing speed 200rpm → 425rpm → 250rpm; 40-72h, air flow 2.0L/min, mixing speed 250rpm.
The significant parameter of different oxygen supply pattern fermenting processs
| Fermentation parameter | The sectional oxygen supply pattern | Single oxygen supply pattern |
| X (dry cell weight)/g.L -1P (anti-microbial activity unit)/U.ml -1Cells produce intensity P X/g.L -1.h -1Product production intensity P P/U.ml -1.h -1Specific cell growth rate μ/h -1Product is than synthesis rate q p/h -1 | 24.22 24.9 0.47 0.52 0.19 0.3 | 19.58 23.2 0.34 0.48 0.23 0.16 |
| Y X/S/g.g -1 Y P/S/U.g -1 | 4.08 4.69 | 3.2 4.31 |
Embodiment 3:
Carry out on the basis that is applied in embodiment 1,2 of Xenorhabdus nematophilus Xenorhabdus nematophilus YL001 fermented liquid:
1. the fermented liquid that obtains in the technology is collected supernatant liquor further at 14000r/min, 4 ℃ of centrifugal 10min;
2, with the PDA substratum that melts with 10 times of supernatant liquor and fermented product dilutions, respectively with the 10ml substratum in the sterilization culture dish of going into diameter 9.0cm, make flat board, be contrast with the PDA that adds aseptic culture medium.From cultivating the plant pathogenic fungi colony edge of 3～6d, be cut into the bacterium piece of diameter 7.5mm with punch tool, place dull and stereotyped central authorities, cultivate 3d down in 20 ℃ of dark conditions, measure each pathogenic bacteria colony diameter;
3, Xenorhabdus nematophilus YL001 fermented product supernatant liquor, 100～300ml (fermented liquid)/L to the red star bacterium of tobacco, tomato early the mycelial growth of vaccine, pumpkin Fusarium oxysporum, cucumber anthrax-bacilus, rice blast fungus and phytophthora blight of pepper stronger restraining effect is arranged, inhibiting rate is 75～100%, wherein fermented product is the highest to the restraining effect of phytophthora blight of pepper mycelial growth, EC
50Be 16.3ml (fermented liquid)/L.
Embodiment 4:
Carry out on the basis that is applied in embodiment 1,2 of Xenorhabdus nematophilus Xenorhabdus nematophilus YL001 fermented liquid:
1, fermented liquid is collected supernatant liquor at 14000r/min, 4 ℃ of centrifugal 10min;
2, the nutrient agar with fusing is cooled to 45 ℃～50 ℃, add for examination pathogenetic bacteria (1.5%V/V) and shake all, every ware (d=9.0cm) is poured the 15ml substratum into, after making flat board, uniformly making a call to three holes with punch tool (diameter 7.5mm) on flat board, add 100 μ l fungal component fermented liquid or fermented products in every hole, is contrast with the substratum, cultivate 24h at 28 ℃, measure antibacterial circle diameter;
3, the supernatant liquor of X.nematophilus YL001 and fermented product have stronger restraining effect (antibacterial circle diameter of fermented liquid, fermented product is respectively 26.50mm, 25.25mm) to streptococcus aureus; Paddy rice bacterial leaf spot bacterium takes second place; To the restraining effect of Salmonellas weak (antibacterial circle diameter of fermented liquid, fermented product is respectively 13.50mm, 13.13mm).
Embodiment 5:
Carry out on the basis that is applied in embodiment 1,2 of the entomopathogenic nematode symbiotic bacteria sterilant of Xenorhabdus nematophilus Xenorhabdus nematophilus YL001 fermented liquid preparation control Phytophthora capsici disease:
Get 65 liters of fermented products, 10025 liters of farming breasts, 10 liters of the 0.5kg/L Sodium Benzoate aqueous solution stir in reactor, promptly get 100 liters of entomopathogenic nematode symbiotic bacteria sterilant.This sterilant is used to prevent and treat the Phytophthora capsici disease.
Embodiment 6:
Present embodiment is got 80 liters of fermented products as different from Example 5,15 liters of sodium dibutyl naphthalene sulfonates, and 5 liters of the 0.5kg/L phenylformic acid aqueous solution stir in reactor, promptly get 100 liters of entomopathogenic nematode symbiotic bacteria sterilant.All the other are with embodiment 5.
Embodiment 7:
Present embodiment is got 65 liters of fermented products as different from Example 5,25 liters of sodium lignosulfonates, and 10 liters of the 0.5kg/L Sorbic Acid aqueous solution stir in reactor, promptly get 100 liters of entomopathogenic nematode symbiotic bacteria sterilant.All the other are with embodiment 5.
Embodiment 8:
Present embodiment is got 70 liters of fermented products as different from Example 5,20 liters of the formaldehyde condensation products of alkyl naphthalenesulfonate, and 10 liters of the 0.5kg/L phenylformic acid aqueous solution stir in reactor, promptly get 100 liters of entomopathogenic nematode symbiotic bacteria sterilant.All the other are with embodiment 5.
Embodiment 9:
Present embodiment is got 65 liters of fermented products as different from Example 5,25 liters of polycarboxylate sodium salts, and 10 liters of the 0.5kg/L Sodium Benzoate aqueous solution stir in reactor, promptly get 100 liters of entomopathogenic nematode symbiotic bacteria sterilant.All the other are with embodiment 5.
The foregoing description can also be enumerated, so long as auxiliary agent well known in the art may be used to prepare the entomopathogenic nematode symbiotic bacteria sterilant of controlling plant diseases.
Embodiment 10:
Carry out on the basis that is applied in embodiment 5～embodiment 9 of Xenorhabdus nematophilus Xenorhabdus nematophilus YL001 fermented product:
From cultivating the phytophthora blight of pepper colony edge of 3～6d, be cut into the bacterium piece of diameter 7.5mm with punch tool, place the flowerpot that contains the vermiculite of sterilizing, with pepper seed with sterilant seed soaking 6 hours after, with twice of flushing with clean water, be sowed on the phytophthora blight of pepper piece that contains in the aseptic vermiculite dixie cup, cover the aseptic vermiculite of one deck then in the above, and watering.With distilled water seed soaking be contrast, 25 ℃ of cultivations, investigated the incidence of seedling respectively at the 12nd day, the diseased plant rate of going down of pepper seedling is respectively 60.6%.
Embodiment 11:
Carry out on the basis that is applied in embodiment 5～embodiment 9 of Xenorhabdus nematophilus Xenorhabdus nematophilus YL001 fermented liquid:
The seed of having sprouted is sowed on the phytophthora blight of pepper piece that contains in the aseptic vermiculite flowerpot, covers the aseptic vermiculite of one deck above, and watering.Cultivating 24h at 25 ℃, is that entomopathogenic nematode symbiotic bacteria sterilant and the 1.7mL/L metaxanin liquid 25mL of 100mL/L waters respectively in flowerpot then with concentration, is contrast with the tap water, is normal healthy controls with what do not contain the phytophthora blight of pepper piece.Cultivate 12d down at 25 ℃～28 ℃, check the incidence of seedling, the diseased plant rate of going down of pepper seedling is respectively 74.34%; The diseased plant rate of going down that metaxanin is handled the back pepper seedling is respectively 64.10%.
Embodiment 12:
Carry out on the basis that is applied in embodiment 5～embodiment 9 of Xenorhabdus nematophilus Xenorhabdus nematophilus YL001 fermented liquid:
Each is prepared by every mu of using dosage and water consumption for the examination sterilant, and clear water is a blank, and every processing repeats 4 times, sub-district of every repetition, and the sub-district area is 15～30m2.Each sub-district is in the field random alignment.
When control tomato and grey mould fruit rot of strawberry, every mu of water 60kg, the control gray mold of cucumber, every mu of water 45kg, dispenser is carried out conventional constant foliar spray with knapsack sprayer.Every 7d dispenser 1 time, dispenser is 4 times altogether.Test method and drug effect statistical method are carried out according to the method for medicine inspecting institute of Ministry of Agriculture promulgation.Prevent and treat that test-results sees Table 1, table 2.
Shaanxi, table 1 embodiment 5 bactericidal agent for preventing and treating graw mold of tomato test of pesticide effectiveness result (fruit) in April, the 2005 Yang Ling Yang Cunnan village
| Medicament | Concentration (ml/L) | After the 2nd dispenser 7 days | After the 4th dispenser 7 days | ||
| Disease index (%) | Preventive effect (%) | Disease index (%) | Preventive effect (%) | ||
| The contrast of embodiment 5 sterilant Sukeling clear water | 200 100 50 500 - | 19.0d 23.0c 30.0b 31.0b 68.0a | 72.06a 66.18b 55.88c 54.41c - | 21.0 25.0 34.0 36.0 82.0 | 75.40a 70.51b 58.54c 56.10d - |
Annotate: with mark same letter person behind the column data, being illustrated on 0.05 level does not have significant difference in 1 table;
2 tests are located at village spring stubble greenhouse tomato field, south, Yang Ling Yangchuan village township, and graw mold of tomato over the years takes place heavier.Soil fertility is medium, pH6.9;
Remove sick fruit before 3 dispensers.
Table 2 embodiment 9 fungicide against Botrytis cinerea on cucumber test of pesticide effectiveness results March three in 2005 former Shaanxi
| Medicament | For examination concentration (ml/L) | 7d after the 2nd dispenser | 7d after the 4th dispenser | ||
| Disease index (%) | Preventive effect (%) | Disease index (%) | Preventive effect (%) | ||
| Embodiment 9 sterilant 40% are executed good happy SC blank | 200 100 50 500 - | 8.32c 9.42b 12.43bc 9.25bc 38.52a | 78.40a 75.55b 67.73c 75.99b - | 15.12d 17.64c 21.45b 17.35c 65.28a | 76.84a 73.88b 67.14c 73.42b - |
Annotate: with mark same letter person behind the column data, being illustrated on 0.05 level does not have significant difference in 1, showing;
2, this test is located at Qu An township, Sanyuan County king Zhuan Cun greenhouse cucumber field, and gray mold over the years takes place heavier.Soil fertility is medium;
Remove sick fruit before 3 dispensers.
Claims (10)
1. the sectional oxygen supply fermentation technology of an entomopathogenic nematode symbiotic bacteria is characterized in that, fermentation strain is Xenorhabdus nematophilus Xenorhabdus nematophilus YL001; And follow these steps to carry out:
(1) kind is guaranteed the above-mentioned Xenorhabdus nematophilus of depositing, lines on the NA culture medium flat plate that cultivate 24～48h for 28 ℃, picking list bacterium colony lines on the NBTA culture medium flat plate again, cultivates 24～48h for 28 ℃;
(2) with the blue colonies on the transfering loop picking NBTA culture medium flat plate, be inoculated in the 250ml triangular flask that 50ml NB substratum is housed, 28 ℃, 180r/min shaking culture 16-24h become primary seed solution;
(3) primary seed solution is equipped with in the 250ml triangular flask of seed culture medium by 9.0% inoculum size access, liquid amount is 25ml, at 26 ℃, 220rpm shaking culture 16-24h, becomes secondary seed solution;
(4) in the 7L stirred-tank fermenter, add the 4.5L fermention medium, 0.01% defoamer, pH7.2, the 0.1-0.15MPa steam sterilizing 30min that offs normal is cooled to 26 ℃, inserts secondary seed solution by 9.0%, cultivate 72h for 26 ℃, fermenting process is regulated air flow and mixing speed, and sectional oxygen supply can obtain fermented product, fermented product is filtered, remove cell and obtain fermented liquid.
2. zymotechnique as claimed in claim 1 is characterized in that, the described seed culture medium of step (3) is: glycerine 5g, yeast extract 15g, 1M-MgSO
45ml, (NH
4)
2SO
42g, 1M-KH
2PO
45ml, 1M-K
2HPO
45ml, 1M-Na
2SO
410ml, water 1000ml, its PH are 7.2.
3. zymotechnique as claimed in claim 1 is characterized in that, the described defoamer of step (4) is a glycerol polyoxyethylene polyoxypropylene ether.
4. zymotechnique as claimed in claim 1 is characterized in that, contains following raw materials in weight in every liter of the described fermention medium of step (4): glucose 6.13g, peptone 21.29g, MgSO
41.50g, (NH
4)
2SO
42.46g, KH
2PO
40.86g, K
2HPO
41.11g, Na
2SO
41.72g.
5. zymotechnique as claimed in claim 1 is characterized in that, the described fermenting process of step (4) regulates air flow and mixing speed sectional oxygen supply pattern is: 0-10h, air flow 1.5L/min, mixing speed 200rpm; 10-40h, air flow 2.5L/min, mixing speed 200rpm → 425rpm → 250rpm; 40-72h, air flow 2.0L/min, mixing speed 250rpm.
6. zymotechnique as claimed in claim 1 is characterized in that, the described NB substratum of step (2) is: extractum carnis 3.0g, peptone 5.0g, water 1000ml, PH7.2～7.4.
7. zymotechnique as claimed in claim 1 is characterized in that, the described NA substratum of step (1) is: extractum carnis 3.0g, peptone 5.0g, nutrient agar medium 20g, water 1000ml, PH7.2～7.4;
The NBTA substratum is: NA substratum+TTC 0.04g+BTB 0.025g.
8. the fermented liquid of the Xenorhabdus nematophilus Xenorhabdus nematophilus YL001 application that is used to prepare the controlling plant diseases sterilant.
9. the entomopathogenic nematode symbiotic bacteria sterilant of a controlling plant diseases, it is characterized in that, this sterilant is made of the volume percent of following component: the fermented liquid 60%-85% of Xenorhabdus nematophilus Xenorhabdusnematophilus YL001, auxiliary agent 1%-5%, surplus is tensio-active agent, and the summation of said components is 100%.
10. sterilant as claimed in claim 9 is characterized in that, described auxiliary agent is a kind of in phenylformic acid, Sodium Benzoate, the Sorbic Acid material, and tensio-active agent is the composition of the known material of this area staff.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CNB2005100428227A CN100370018C (en) | 2005-06-16 | 2005-06-16 | Segmented oxygen supply fermentation process of entomopathogenic nematode symbiotic bacteria and use of fermentation product thereof |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CNB2005100428227A CN100370018C (en) | 2005-06-16 | 2005-06-16 | Segmented oxygen supply fermentation process of entomopathogenic nematode symbiotic bacteria and use of fermentation product thereof |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN1724651A CN1724651A (en) | 2006-01-25 |
| CN100370018C true CN100370018C (en) | 2008-02-20 |
Family
ID=35924281
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CNB2005100428227A Expired - Fee Related CN100370018C (en) | 2005-06-16 | 2005-06-16 | Segmented oxygen supply fermentation process of entomopathogenic nematode symbiotic bacteria and use of fermentation product thereof |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN100370018C (en) |
Families Citing this family (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN100415873C (en) * | 2006-10-17 | 2008-09-03 | 广东省昆虫研究所 | Bacteria strain (X-7 strain) for culturing bait eelworm and its eelworm culture method |
| CN101892170B (en) * | 2009-05-20 | 2012-07-04 | 河北农业大学 | Entomopathogenic nematode symbiotic bacteria and application thereof |
| CN111647522B (en) * | 2020-05-06 | 2022-04-29 | 南京农业大学 | Entomopathogenic nematode symbiotic bacterial strain for inhibiting various plant fungal diseases and application thereof |
| CN114304146A (en) * | 2021-12-07 | 2022-04-12 | 西北农林科技大学 | Xcn 1-containing microbial source sterilization ointment, and preparation method and application thereof |
Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1233938A (en) * | 1996-08-29 | 1999-11-03 | 英王陛下的大不列颠及北爱尔兰联合王国政府农业渔业和食品部 | Pesticidal agent |
-
2005
- 2005-06-16 CN CNB2005100428227A patent/CN100370018C/en not_active Expired - Fee Related
Patent Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1233938A (en) * | 1996-08-29 | 1999-11-03 | 英王陛下的大不列颠及北爱尔兰联合王国政府农业渔业和食品部 | Pesticidal agent |
Non-Patent Citations (2)
| Title |
|---|
| 昆虫病原线虫共生菌培养与活性研究. 王永宏.西北农林科技大学博士学位论文. 2004 * |
| 高毒力昆虫病原线虫共生细菌菌株的筛选及其杀虫活性的检测. 王立霞等.昆虫学报，第43卷第增刊期. 2000 * |
Also Published As
| Publication number | Publication date |
|---|---|
| CN1724651A (en) | 2006-01-25 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| Elshahawy et al. | First report of Pythium aphanidermatum infecting tomato in Egypt and its control using biogenic silver nanoparticles | |
| KR101334742B1 (en) | A novel Bacillus subtilis strain and use thereof for protecting the rust of plant roots | |
| CN101473853B (en) | Application of Bacillus cereus Bacillus cereusCMCC63305 in agriculture field | |
| CN104388356A (en) | Streptomyces sampsonii strain as well as separation method and application thereof | |
| KR101184296B1 (en) | Composition for anti plant pathogen containing bacillus subtilis kbc1010 or its culture fluid as an active ingredient | |
| CN1951197A (en) | A novel method for controlling soybean root disease by using rhizobia | |
| BG67257B1 (en) | Bacterial strain of bacillus amyloliquefaciens subsp. plantarum bs89 as a means of increasing plant productivity and their protection against diseases | |
| CN103975955A (en) | Composite biological seed coating agent used for controlling vegetable soil-borne diseases and preparation and application methods thereof | |
| CN1724649A (en) | Entomopathogenic nematode symbiotic bacteria fermentation process and application of fermentation liquor thereof | |
| CN102643148A (en) | Special biological disease-resistant vermin-proof culture medium for flowers and preparation method thereof | |
| CN101225368A (en) | Biocontrol actinomyces-streptomyces diastatochromogenes D | |
| KR20140071145A (en) | Novel Paenibacillus polymyxa AB-15 strain and use the same | |
| KR20120068674A (en) | Streptomyces griseus big105 for controlling plant diseases and uses thereof | |
| CN100370018C (en) | Segmented oxygen supply fermentation process of entomopathogenic nematode symbiotic bacteria and use of fermentation product thereof | |
| CN103141519A (en) | Preparation method and application of anti-biological inoculants for controlling crop soil-borne diseases | |
| KR101175532B1 (en) | Novel burkholderia sp., and media for mass culture of burkholderia for increasing antifungal effect and method for mass production thereof using the same | |
| CN100364411C (en) | Microbiological agent resisting rice blast, preparation method and application thereof | |
| KR20170054158A (en) | Burkholderia ambifaria CJ4 strain Possessing antifungal activity against Root Rot Pathogen of ginseng and Use Thereof | |
| CN108314522A (en) | A kind of disease-resisting, insecticidal biological organic fertilizer and preparation method thereof | |
| CN101008017A (en) | Bacillus subtilis (Ehrenberg)Cohn fermented product and its production method and uses | |
| CN102443559B (en) | Baclillus subtilis used for controlling cotton verticillium wilt and application thereof | |
| KR20020031961A (en) | Streptomyces kasugaensis GBA-0927 | |
| KR101279040B1 (en) | Exiguobacterium acetylicum WCU292 strain, composition for control plant disease and control method of plant disease with same | |
| JP2003531603A (en) | Microbial preparation for biological control using novel Trichoderma microorganism strain and method for producing the same | |
| CN108587979B (en) | Bacillus subtilis and application thereof |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| C14 | Grant of patent or utility model | ||
| GR01 | Patent grant | ||
| ASS | Succession or assignment of patent right |
Owner name: FUJI BIOTECH (SHANGHAI) CO., LTD. Free format text: FORMER OWNER: NUISANCELESS PESTICIDE RESEARCH SERVICE CENTER, XIBEI AGROFORESTRY SCIENCE AND T Effective date: 20121127 |
|
| C41 | Transfer of patent application or patent right or utility model | ||
| COR | Change of bibliographic data |
Free format text: CORRECT: ADDRESS; FROM: 712100 XIANYANG, SHAANXI PROVINCE TO: 200443 BAOSHAN, SHANGHAI |
|
| TR01 | Transfer of patent right |
Effective date of registration: 20121127 Address after: 200443 Shanghai city Baoshan District Ji Yunlu No. 588 South 10C Patentee after: FUJI Biotech (Shanghai) Co., Ltd. Address before: 712100 Shaanxi province Yangling District Tai Road No. 3 Patentee before: Nuisanceless Pesticide Research Service Center, Xibei Agroforestry Science and T |
|
| 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: 20080220 Termination date: 20180616 |