CN108570433A - One plant of acid proof defence pseudomonad CLP-6 and its application - Google Patents
One plant of acid proof defence pseudomonad CLP-6 and its application Download PDFInfo
- Publication number
- CN108570433A CN108570433A CN201810438730.8A CN201810438730A CN108570433A CN 108570433 A CN108570433 A CN 108570433A CN 201810438730 A CN201810438730 A CN 201810438730A CN 108570433 A CN108570433 A CN 108570433A
- Authority
- CN
- China
- Prior art keywords
- clp
- microbial
- bacterial
- pseudomonad
- tobacco
- 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
- 239000002253 acid Substances 0.000 title claims abstract description 43
- 230000001580 bacterial effect Effects 0.000 claims abstract description 79
- 241000208125 Nicotiana Species 0.000 claims abstract description 69
- 235000002637 Nicotiana tabacum Nutrition 0.000 claims abstract description 69
- 230000000813 microbial effect Effects 0.000 claims abstract description 29
- 241000196324 Embryophyta Species 0.000 claims abstract description 25
- 244000005700 microbiome Species 0.000 claims abstract description 7
- 241000894006 Bacteria Species 0.000 claims description 54
- 241000467496 Pseudomonas protegens Species 0.000 claims description 30
- 239000003795 chemical substances by application Substances 0.000 claims description 28
- ZLMJMSJWJFRBEC-UHFFFAOYSA-N Potassium Chemical compound [K] ZLMJMSJWJFRBEC-UHFFFAOYSA-N 0.000 claims description 22
- 239000011591 potassium Substances 0.000 claims description 22
- 229910052700 potassium Inorganic materials 0.000 claims description 22
- 239000012530 fluid Substances 0.000 claims description 21
- 239000003337 fertilizer Substances 0.000 claims description 20
- 238000000034 method Methods 0.000 claims description 14
- 235000015097 nutrients Nutrition 0.000 claims description 8
- 208000031888 Mycoses Diseases 0.000 claims description 7
- 230000006806 disease prevention Effects 0.000 claims description 7
- IJGRMHOSHXDMSA-UHFFFAOYSA-N Atomic nitrogen Chemical compound N#N IJGRMHOSHXDMSA-UHFFFAOYSA-N 0.000 claims description 6
- 241000223602 Alternaria alternata Species 0.000 claims description 5
- 230000001737 promoting effect Effects 0.000 claims description 4
- 239000002552 dosage form Substances 0.000 claims description 3
- 229910052757 nitrogen Inorganic materials 0.000 claims description 3
- 239000005416 organic matter Substances 0.000 claims description 3
- 239000000375 suspending agent Substances 0.000 claims description 3
- 239000004562 water dispersible granule Substances 0.000 claims description 3
- 239000004563 wettable powder Substances 0.000 claims description 3
- 239000007900 aqueous suspension Substances 0.000 claims description 2
- 239000002689 soil Substances 0.000 abstract description 27
- 201000010099 disease Diseases 0.000 abstract description 24
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 abstract description 24
- 230000003042 antagnostic effect Effects 0.000 abstract description 20
- 230000001717 pathogenic effect Effects 0.000 abstract description 19
- 244000052769 pathogen Species 0.000 abstract description 18
- 230000000694 effects Effects 0.000 abstract description 17
- JEIPFZHSYJVQDO-UHFFFAOYSA-N iron(III) oxide Inorganic materials O=[Fe]O[Fe]=O JEIPFZHSYJVQDO-UHFFFAOYSA-N 0.000 abstract description 12
- 230000002265 prevention Effects 0.000 abstract description 10
- 230000000443 biocontrol Effects 0.000 abstract description 5
- 238000011160 research Methods 0.000 abstract description 3
- 230000000845 anti-microbial effect Effects 0.000 abstract description 2
- 238000011161 development Methods 0.000 abstract description 2
- 230000020477 pH reduction Effects 0.000 abstract description 2
- 229930000044 secondary metabolite Natural products 0.000 abstract description 2
- 238000001228 spectrum Methods 0.000 abstract description 2
- 239000006228 supernatant Substances 0.000 description 25
- 241000589771 Ralstonia solanacearum Species 0.000 description 24
- 241000233647 Phytophthora nicotianae var. parasitica Species 0.000 description 22
- 239000000243 solution Substances 0.000 description 21
- 229910019142 PO4 Inorganic materials 0.000 description 14
- 230000006378 damage Effects 0.000 description 14
- 239000010452 phosphate Substances 0.000 description 14
- 239000001963 growth medium Substances 0.000 description 13
- NBIIXXVUZAFLBC-UHFFFAOYSA-K phosphate Chemical compound [O-]P([O-])([O-])=O NBIIXXVUZAFLBC-UHFFFAOYSA-K 0.000 description 13
- 238000012360 testing method Methods 0.000 description 13
- OAICVXFJPJFONN-UHFFFAOYSA-N Phosphorus Chemical compound [P] OAICVXFJPJFONN-UHFFFAOYSA-N 0.000 description 12
- 235000019504 cigarettes Nutrition 0.000 description 12
- 229910052698 phosphorus Inorganic materials 0.000 description 12
- 239000011574 phosphorus Substances 0.000 description 12
- FRXSZNDVFUDTIR-UHFFFAOYSA-N 6-methoxy-1,2,3,4-tetrahydroquinoline Chemical compound N1CCCC2=CC(OC)=CC=C21 FRXSZNDVFUDTIR-UHFFFAOYSA-N 0.000 description 11
- -1 antifreeze Substances 0.000 description 10
- 239000007788 liquid Substances 0.000 description 10
- 238000012545 processing Methods 0.000 description 9
- 230000002401 inhibitory effect Effects 0.000 description 8
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 8
- 239000000463 material Substances 0.000 description 7
- 239000000575 pesticide Substances 0.000 description 7
- 241000193830 Bacillus <bacterium> Species 0.000 description 6
- 239000001888 Peptone Substances 0.000 description 6
- 108010080698 Peptones Proteins 0.000 description 6
- 230000012010 growth Effects 0.000 description 6
- 235000019319 peptone Nutrition 0.000 description 6
- 238000005063 solubilization Methods 0.000 description 6
- 230000007928 solubilization Effects 0.000 description 6
- 239000000725 suspension Substances 0.000 description 6
- 108090000790 Enzymes Proteins 0.000 description 5
- 102000004190 Enzymes Human genes 0.000 description 5
- ISWSIDIOOBJBQZ-UHFFFAOYSA-N Phenol Natural products OC1=CC=CC=C1 ISWSIDIOOBJBQZ-UHFFFAOYSA-N 0.000 description 5
- VYPSYNLAJGMNEJ-UHFFFAOYSA-N Silicium dioxide Chemical compound O=[Si]=O VYPSYNLAJGMNEJ-UHFFFAOYSA-N 0.000 description 5
- 235000015278 beef Nutrition 0.000 description 5
- 239000003153 chemical reaction reagent Substances 0.000 description 5
- 239000002270 dispersing agent Substances 0.000 description 5
- 238000002156 mixing Methods 0.000 description 5
- 241000233866 Fungi Species 0.000 description 4
- 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 4
- 108091005804 Peptidases Proteins 0.000 description 4
- 241000589516 Pseudomonas Species 0.000 description 4
- XSQUKJJJFZCRTK-UHFFFAOYSA-N Urea Chemical compound NC(N)=O XSQUKJJJFZCRTK-UHFFFAOYSA-N 0.000 description 4
- WQZGKKKJIJFFOK-VFUOTHLCSA-N beta-D-glucose Chemical compound OC[C@H]1O[C@@H](O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-VFUOTHLCSA-N 0.000 description 4
- 230000005764 inhibitory process Effects 0.000 description 4
- 239000002609 medium Substances 0.000 description 4
- 238000009335 monocropping Methods 0.000 description 4
- 239000000843 powder Substances 0.000 description 4
- 238000001556 precipitation Methods 0.000 description 4
- QTBSBXVTEAMEQO-UHFFFAOYSA-N Acetic acid Chemical compound CC(O)=O QTBSBXVTEAMEQO-UHFFFAOYSA-N 0.000 description 3
- LYCAIKOWRPUZTN-UHFFFAOYSA-N Ethylene glycol Chemical compound OCCO LYCAIKOWRPUZTN-UHFFFAOYSA-N 0.000 description 3
- WSFSSNUMVMOOMR-UHFFFAOYSA-N Formaldehyde Chemical compound O=C WSFSSNUMVMOOMR-UHFFFAOYSA-N 0.000 description 3
- 102000035195 Peptidases Human genes 0.000 description 3
- DNIAPMSPPWPWGF-UHFFFAOYSA-N Propylene glycol Chemical compound CC(O)CO DNIAPMSPPWPWGF-UHFFFAOYSA-N 0.000 description 3
- 241000607479 Yersinia pestis Species 0.000 description 3
- 238000002835 absorbance Methods 0.000 description 3
- 230000002528 anti-freeze Effects 0.000 description 3
- 239000002518 antifoaming agent Substances 0.000 description 3
- 230000003385 bacteriostatic effect Effects 0.000 description 3
- 239000011230 binding agent Substances 0.000 description 3
- 238000009395 breeding Methods 0.000 description 3
- 230000001488 breeding effect Effects 0.000 description 3
- 210000004027 cell Anatomy 0.000 description 3
- 238000005119 centrifugation Methods 0.000 description 3
- 239000007884 disintegrant Substances 0.000 description 3
- 230000003628 erosive effect Effects 0.000 description 3
- 238000000855 fermentation Methods 0.000 description 3
- 230000004151 fermentation Effects 0.000 description 3
- 239000000945 filler Substances 0.000 description 3
- 239000008103 glucose Substances 0.000 description 3
- 230000007935 neutral effect Effects 0.000 description 3
- 235000019833 protease Nutrition 0.000 description 3
- 239000002904 solvent Substances 0.000 description 3
- 239000002562 thickening agent Substances 0.000 description 3
- 239000000080 wetting agent Substances 0.000 description 3
- 108020004465 16S ribosomal RNA Proteins 0.000 description 2
- 241000589291 Acinetobacter Species 0.000 description 2
- 241000228212 Aspergillus Species 0.000 description 2
- 241000588914 Enterobacter Species 0.000 description 2
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 description 2
- 241000187809 Frankia Species 0.000 description 2
- PEDCQBHIVMGVHV-UHFFFAOYSA-N Glycerine Chemical compound OCC(O)CO PEDCQBHIVMGVHV-UHFFFAOYSA-N 0.000 description 2
- 238000002768 Kirby-Bauer method Methods 0.000 description 2
- TWRXJAOTZQYOKJ-UHFFFAOYSA-L Magnesium chloride Chemical compound [Mg+2].[Cl-].[Cl-] TWRXJAOTZQYOKJ-UHFFFAOYSA-L 0.000 description 2
- 241000233645 Phytophthora nicotianae Species 0.000 description 2
- 239000002202 Polyethylene glycol Substances 0.000 description 2
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 description 2
- 241000122971 Stenotrophomonas Species 0.000 description 2
- 241000223259 Trichoderma Species 0.000 description 2
- WYWFMUBFNXLFJK-UHFFFAOYSA-N [Mo].[Sb] Chemical compound [Mo].[Sb] WYWFMUBFNXLFJK-UHFFFAOYSA-N 0.000 description 2
- VSCWAEJMTAWNJL-UHFFFAOYSA-K aluminium trichloride Chemical compound Cl[Al](Cl)Cl VSCWAEJMTAWNJL-UHFFFAOYSA-K 0.000 description 2
- 230000008485 antagonism Effects 0.000 description 2
- 244000052616 bacterial pathogen Species 0.000 description 2
- 239000000440 bentonite Substances 0.000 description 2
- 229910000278 bentonite Inorganic materials 0.000 description 2
- SVPXDRXYRYOSEX-UHFFFAOYSA-N bentoquatam Chemical compound O.O=[Si]=O.O=[Al]O[Al]=O SVPXDRXYRYOSEX-UHFFFAOYSA-N 0.000 description 2
- 239000004202 carbamide Substances 0.000 description 2
- 239000005018 casein Substances 0.000 description 2
- 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 2
- 235000021240 caseins Nutrition 0.000 description 2
- 238000006243 chemical reaction Methods 0.000 description 2
- 239000013065 commercial product Substances 0.000 description 2
- 230000001066 destructive effect Effects 0.000 description 2
- 238000004821 distillation Methods 0.000 description 2
- 239000012153 distilled water Substances 0.000 description 2
- 235000019441 ethanol Nutrition 0.000 description 2
- 238000002474 experimental method Methods 0.000 description 2
- 230000001408 fungistatic effect Effects 0.000 description 2
- 230000003450 growing effect Effects 0.000 description 2
- 101150013736 gyrB gene Proteins 0.000 description 2
- 238000004519 manufacturing process Methods 0.000 description 2
- 238000000386 microscopy Methods 0.000 description 2
- 230000000877 morphologic effect Effects 0.000 description 2
- 239000004570 mortar (masonry) Substances 0.000 description 2
- 230000000474 nursing effect Effects 0.000 description 2
- 230000010355 oscillation Effects 0.000 description 2
- 229920001223 polyethylene glycol Polymers 0.000 description 2
- 239000010453 quartz Substances 0.000 description 2
- 239000011734 sodium Substances 0.000 description 2
- 239000008223 sterile water Substances 0.000 description 2
- 239000001038 titanium pigment Substances 0.000 description 2
- YNJBWRMUSHSURL-UHFFFAOYSA-N trichloroacetic acid Chemical compound OC(=O)C(Cl)(Cl)Cl YNJBWRMUSHSURL-UHFFFAOYSA-N 0.000 description 2
- DWNBOPVKNPVNQG-LURJTMIESA-N (2s)-4-hydroxy-2-(propylamino)butanoic acid Chemical compound CCCN[C@H](C(O)=O)CCO DWNBOPVKNPVNQG-LURJTMIESA-N 0.000 description 1
- RSWGJHLUYNHPMX-UHFFFAOYSA-N Abietic-Saeure Natural products C12CCC(C(C)C)=CC2=CCC2C1(C)CCCC2(C)C(O)=O RSWGJHLUYNHPMX-UHFFFAOYSA-N 0.000 description 1
- 241000223600 Alternaria Species 0.000 description 1
- 241000186063 Arthrobacter Species 0.000 description 1
- LSNNMFCWUKXFEE-UHFFFAOYSA-M Bisulfite Chemical compound OS([O-])=O LSNNMFCWUKXFEE-UHFFFAOYSA-M 0.000 description 1
- 241000555281 Brevibacillus Species 0.000 description 1
- VTYYLEPIZMXCLO-UHFFFAOYSA-L Calcium carbonate Chemical compound [Ca+2].[O-]C([O-])=O VTYYLEPIZMXCLO-UHFFFAOYSA-L 0.000 description 1
- 244000025254 Cannabis sativa Species 0.000 description 1
- 229920002134 Carboxymethyl cellulose Polymers 0.000 description 1
- 229920000858 Cyclodextrin Polymers 0.000 description 1
- 208000035240 Disease Resistance Diseases 0.000 description 1
- 239000004593 Epoxy Substances 0.000 description 1
- 108010010803 Gelatin Proteins 0.000 description 1
- 241000626621 Geobacillus Species 0.000 description 1
- 206010053759 Growth retardation Diseases 0.000 description 1
- 208000025309 Hair disease Diseases 0.000 description 1
- DGAQECJNVWCQMB-PUAWFVPOSA-M Ilexoside XXIX Chemical compound C[C@@H]1CC[C@@]2(CC[C@@]3(C(=CC[C@H]4[C@]3(CC[C@@H]5[C@@]4(CC[C@@H](C5(C)C)OS(=O)(=O)[O-])C)C)[C@@H]2[C@]1(C)O)C)C(=O)O[C@H]6[C@@H]([C@H]([C@@H]([C@H](O6)CO)O)O)O.[Na+] DGAQECJNVWCQMB-PUAWFVPOSA-M 0.000 description 1
- 241000568397 Lysinibacillus Species 0.000 description 1
- RCEAADKTGXTDOA-UHFFFAOYSA-N OS(O)(=O)=O.CCCCCCCCCCCC[Na] Chemical compound OS(O)(=O)=O.CCCCCCCCCCCC[Na] RCEAADKTGXTDOA-UHFFFAOYSA-N 0.000 description 1
- 241000896292 Odontothrips loti Species 0.000 description 1
- 241000179039 Paenibacillus Species 0.000 description 1
- 244000131316 Panax pseudoginseng Species 0.000 description 1
- 235000005035 Panax pseudoginseng ssp. pseudoginseng Nutrition 0.000 description 1
- 235000003140 Panax quinquefolius Nutrition 0.000 description 1
- 241000228143 Penicillium Species 0.000 description 1
- 241000233629 Phytophthora parasitica Species 0.000 description 1
- 229920003171 Poly (ethylene oxide) Polymers 0.000 description 1
- 239000004372 Polyvinyl alcohol Substances 0.000 description 1
- 239000004365 Protease Substances 0.000 description 1
- 102100037486 Reverse transcriptase/ribonuclease H Human genes 0.000 description 1
- KHPCPRHQVVSZAH-HUOMCSJISA-N Rosin Natural products O(C/C=C/c1ccccc1)[C@H]1[C@H](O)[C@@H](O)[C@@H](O)[C@@H](CO)O1 KHPCPRHQVVSZAH-HUOMCSJISA-N 0.000 description 1
- VMHLLURERBWHNL-UHFFFAOYSA-M Sodium acetate Chemical compound [Na+].CC([O-])=O VMHLLURERBWHNL-UHFFFAOYSA-M 0.000 description 1
- 244000061456 Solanum tuberosum Species 0.000 description 1
- 235000002595 Solanum tuberosum Nutrition 0.000 description 1
- 229920002472 Starch Polymers 0.000 description 1
- 241000187747 Streptomyces Species 0.000 description 1
- 241001655322 Streptomycetales Species 0.000 description 1
- QAOWNCQODCNURD-UHFFFAOYSA-L Sulfate Chemical compound [O-]S([O-])(=O)=O QAOWNCQODCNURD-UHFFFAOYSA-L 0.000 description 1
- 244000269722 Thea sinensis Species 0.000 description 1
- 241001002356 Valeriana edulis Species 0.000 description 1
- 230000002378 acidificating effect Effects 0.000 description 1
- 230000002776 aggregation Effects 0.000 description 1
- 238000004220 aggregation Methods 0.000 description 1
- BFNBIHQBYMNNAN-UHFFFAOYSA-N ammonium sulfate Chemical compound N.N.OS(O)(=O)=O BFNBIHQBYMNNAN-UHFFFAOYSA-N 0.000 description 1
- 229910052921 ammonium sulfate Inorganic materials 0.000 description 1
- 235000011130 ammonium sulphate Nutrition 0.000 description 1
- 125000000129 anionic group Chemical group 0.000 description 1
- 230000000844 anti-bacterial effect Effects 0.000 description 1
- 229960000892 attapulgite Drugs 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 239000001768 carboxy methyl cellulose Substances 0.000 description 1
- 235000010948 carboxy methyl cellulose Nutrition 0.000 description 1
- 229920003064 carboxyethyl cellulose Polymers 0.000 description 1
- 150000007942 carboxylates Chemical class 0.000 description 1
- 239000008112 carboxymethyl-cellulose Substances 0.000 description 1
- 210000000170 cell membrane Anatomy 0.000 description 1
- 210000002421 cell wall Anatomy 0.000 description 1
- 230000008859 change Effects 0.000 description 1
- 238000004737 colorimetric analysis Methods 0.000 description 1
- 150000001875 compounds Chemical class 0.000 description 1
- 239000007859 condensation product Substances 0.000 description 1
- 230000007797 corrosion Effects 0.000 description 1
- 238000005260 corrosion Methods 0.000 description 1
- 210000000805 cytoplasm Anatomy 0.000 description 1
- 239000008367 deionised water Substances 0.000 description 1
- 229910021641 deionized water Inorganic materials 0.000 description 1
- 239000008121 dextrose Substances 0.000 description 1
- 238000009792 diffusion process Methods 0.000 description 1
- 238000007865 diluting Methods 0.000 description 1
- 238000010790 dilution Methods 0.000 description 1
- 239000012895 dilution Substances 0.000 description 1
- 208000034653 disorder of pilosebaceous unit Diseases 0.000 description 1
- 238000009826 distribution Methods 0.000 description 1
- 238000001035 drying Methods 0.000 description 1
- QYDYPVFESGNLHU-UHFFFAOYSA-N elaidic acid methyl ester Natural products CCCCCCCCC=CCCCCCCCC(=O)OC QYDYPVFESGNLHU-UHFFFAOYSA-N 0.000 description 1
- 238000005516 engineering process Methods 0.000 description 1
- 230000002255 enzymatic effect Effects 0.000 description 1
- 210000003608 fece Anatomy 0.000 description 1
- NVVZQXQBYZPMLJ-UHFFFAOYSA-N formaldehyde;naphthalene-1-sulfonic acid Chemical compound O=C.C1=CC=C2C(S(=O)(=O)O)=CC=CC2=C1 NVVZQXQBYZPMLJ-UHFFFAOYSA-N 0.000 description 1
- 229920000159 gelatin Polymers 0.000 description 1
- 239000008273 gelatin Substances 0.000 description 1
- 235000019322 gelatine Nutrition 0.000 description 1
- 235000011852 gelatine desserts Nutrition 0.000 description 1
- 235000008434 ginseng Nutrition 0.000 description 1
- 235000011187 glycerol Nutrition 0.000 description 1
- 235000013882 gravy Nutrition 0.000 description 1
- 231100000001 growth retardation Toxicity 0.000 description 1
- 238000011081 inoculation Methods 0.000 description 1
- 230000001788 irregular Effects 0.000 description 1
- 238000002955 isolation Methods 0.000 description 1
- 238000002386 leaching Methods 0.000 description 1
- 230000031700 light absorption Effects 0.000 description 1
- 229920005610 lignin Polymers 0.000 description 1
- 230000000670 limiting effect Effects 0.000 description 1
- 239000010871 livestock manure Substances 0.000 description 1
- 229910001629 magnesium chloride Inorganic materials 0.000 description 1
- 239000011159 matrix material Substances 0.000 description 1
- 238000005259 measurement Methods 0.000 description 1
- QYDYPVFESGNLHU-KHPPLWFESA-N methyl oleate Chemical compound CCCCCCCC\C=C/CCCCCCCC(=O)OC QYDYPVFESGNLHU-KHPPLWFESA-N 0.000 description 1
- 229940073769 methyl oleate Drugs 0.000 description 1
- 239000000203 mixture Substances 0.000 description 1
- 230000003020 moisturizing effect Effects 0.000 description 1
- LCRMGUFGEDUSOG-UHFFFAOYSA-N naphthalen-1-ylsulfonyloxymethyl naphthalene-1-sulfonate;sodium Chemical compound [Na].C1=CC=C2C(S(=O)(OCOS(=O)(=O)C=3C4=CC=CC=C4C=CC=3)=O)=CC=CC2=C1 LCRMGUFGEDUSOG-UHFFFAOYSA-N 0.000 description 1
- 235000016709 nutrition Nutrition 0.000 description 1
- 230000035764 nutrition Effects 0.000 description 1
- 239000003921 oil Substances 0.000 description 1
- 235000019198 oils Nutrition 0.000 description 1
- 229910052625 palygorskite Inorganic materials 0.000 description 1
- 101150012629 parE gene Proteins 0.000 description 1
- 229920005646 polycarboxylate Polymers 0.000 description 1
- 229920001296 polysiloxane Polymers 0.000 description 1
- 229920002451 polyvinyl alcohol Polymers 0.000 description 1
- 229940088417 precipitated calcium carbonate Drugs 0.000 description 1
- 238000004321 preservation Methods 0.000 description 1
- 230000003449 preventive effect Effects 0.000 description 1
- 230000008569 process Effects 0.000 description 1
- 235000019419 proteases Nutrition 0.000 description 1
- 108090000623 proteins and genes Proteins 0.000 description 1
- 230000009467 reduction Effects 0.000 description 1
- 230000000452 restraining effect Effects 0.000 description 1
- 238000005070 sampling Methods 0.000 description 1
- 235000021003 saturated fats Nutrition 0.000 description 1
- HFHDHCJBZVLPGP-UHFFFAOYSA-N schardinger α-dextrin Chemical compound O1C(C(C2O)O)C(CO)OC2OC(C(C2O)O)C(CO)OC2OC(C(C2O)O)C(CO)OC2OC(C(O)C2O)C(CO)OC2OC(C(C2O)O)C(CO)OC2OC2C(O)C(O)C1OC2CO HFHDHCJBZVLPGP-UHFFFAOYSA-N 0.000 description 1
- 238000012216 screening Methods 0.000 description 1
- 230000028327 secretion Effects 0.000 description 1
- 229920002545 silicone oil Polymers 0.000 description 1
- 229910052708 sodium Inorganic materials 0.000 description 1
- 239000001632 sodium acetate Substances 0.000 description 1
- 235000017281 sodium acetate Nutrition 0.000 description 1
- 239000011780 sodium chloride Substances 0.000 description 1
- PYODKQIVQIVELM-UHFFFAOYSA-M sodium;2,3-bis(2-methylpropyl)naphthalene-1-sulfonate Chemical compound [Na+].C1=CC=C2C(S([O-])(=O)=O)=C(CC(C)C)C(CC(C)C)=CC2=C1 PYODKQIVQIVELM-UHFFFAOYSA-M 0.000 description 1
- 230000003381 solubilizing effect Effects 0.000 description 1
- 235000012424 soybean oil Nutrition 0.000 description 1
- 239000003549 soybean oil Substances 0.000 description 1
- 239000008107 starch Substances 0.000 description 1
- 235000019698 starch Nutrition 0.000 description 1
- 230000001954 sterilising effect Effects 0.000 description 1
- 239000000126 substance Substances 0.000 description 1
- 230000001629 suppression Effects 0.000 description 1
- 230000008961 swelling Effects 0.000 description 1
- 208000024891 symptom Diseases 0.000 description 1
- KHPCPRHQVVSZAH-UHFFFAOYSA-N trans-cinnamyl beta-D-glucopyranoside Natural products OC1C(O)C(O)C(CO)OC1OCC=CC1=CC=CC=C1 KHPCPRHQVVSZAH-UHFFFAOYSA-N 0.000 description 1
- 241001515965 unidentified phage Species 0.000 description 1
- 230000002792 vascular Effects 0.000 description 1
- 238000012795 verification Methods 0.000 description 1
- 239000010455 vermiculite Substances 0.000 description 1
- 229910052902 vermiculite Inorganic materials 0.000 description 1
- 235000019354 vermiculite Nutrition 0.000 description 1
- 229920001285 xanthan gum Polymers 0.000 description 1
Classifications
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
- C12N1/20—Bacteria; Culture media therefor
- C12N1/205—Bacterial isolates
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12R—INDEXING SCHEME ASSOCIATED WITH SUBCLASSES C12C - C12Q, RELATING TO MICROORGANISMS
- C12R2001/00—Microorganisms ; Processes using microorganisms
- C12R2001/01—Bacteria or Actinomycetales ; using bacteria or Actinomycetales
- C12R2001/38—Pseudomonas
-
- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01N—PRESERVATION OF BODIES OF HUMANS OR ANIMALS OR PLANTS OR PARTS THEREOF; BIOCIDES, e.g. AS DISINFECTANTS, AS PESTICIDES OR AS HERBICIDES; PEST REPELLANTS OR ATTRACTANTS; PLANT GROWTH REGULATORS
- A01N63/00—Biocides, pest repellants or attractants, or plant growth regulators containing microorganisms, viruses, microbial fungi, animals or substances produced by, or obtained from, microorganisms, viruses, microbial fungi or animals, e.g. enzymes or fermentates
-
- C—CHEMISTRY; METALLURGY
- C05—FERTILISERS; MANUFACTURE THEREOF
- C05F—ORGANIC FERTILISERS NOT COVERED BY SUBCLASSES C05B, C05C, e.g. FERTILISERS FROM WASTE OR REFUSE
- C05F11/00—Other organic fertilisers
- C05F11/08—Organic fertilisers containing added bacterial cultures, mycelia or the like
-
- C—CHEMISTRY; METALLURGY
- C05—FERTILISERS; MANUFACTURE THEREOF
- C05G—MIXTURES OF FERTILISERS COVERED INDIVIDUALLY BY DIFFERENT SUBCLASSES OF CLASS C05; MIXTURES OF ONE OR MORE FERTILISERS WITH MATERIALS NOT HAVING A SPECIFIC FERTILISING ACTIVITY, e.g. PESTICIDES, SOIL-CONDITIONERS, WETTING AGENTS; FERTILISERS CHARACTERISED BY THEIR FORM
- C05G3/00—Mixtures of one or more fertilisers with additives not having a specially fertilising activity
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
- C12N1/20—Bacteria; Culture media therefor
Landscapes
- Life Sciences & Earth Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Health & Medical Sciences (AREA)
- Engineering & Computer Science (AREA)
- Organic Chemistry (AREA)
- Zoology (AREA)
- Wood Science & Technology (AREA)
- Biotechnology (AREA)
- Bioinformatics & Cheminformatics (AREA)
- General Health & Medical Sciences (AREA)
- Genetics & Genomics (AREA)
- Microbiology (AREA)
- Biochemistry (AREA)
- General Engineering & Computer Science (AREA)
- Virology (AREA)
- Tropical Medicine & Parasitology (AREA)
- Biomedical Technology (AREA)
- Medicinal Chemistry (AREA)
- Pest Control & Pesticides (AREA)
- Agronomy & Crop Science (AREA)
- Plant Pathology (AREA)
- Dentistry (AREA)
- Environmental Sciences (AREA)
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
- Agricultural Chemicals And Associated Chemicals (AREA)
Abstract
The present invention provides one plant of acid proof defence pseudomonad CLP 6 and its application.The bacterial strain is preserved in China Committee for Culture Collection of Microorganisms's common micro-organisms center on October 27th, 2016, and biological deposits number are CGMCC No.13205.The bacterial strain has acid resistance, and diseases prevention, growth-promoting effect are all good, and resistance is strong, which has both pathogen antagonistic activity, and it is wide to the acidity tolerance range of soil, antimicrobial spectrum is wide, have application and development potentiality in the prevention of tobacco bacterial wilt, balck shank and rust.Simultaneously, under the increasingly severe overall background of China's acidification situation, the research of the bacterial strain also has reference for the control of plant disease under various soil properties, microbial resources are provided for the green prevention and control of tobacco bacterial wilt, balck shank and rust in acid soil, microbial secondary metabolites is excavated and then improves bacterial wilt, balck shank and rust biocontrol effect in acid soil and establish important foundation.
Description
Technical field
The invention belongs to microorganisms technical fields, and in particular to one plant of acid proof defence pseudomonad CLP-6 and its answer
With.
Background technology
Tobacco can be caused harm in entire breeding time by a variety of fungies and bacteriosis, domestic tobacco diseases 60 on the books
Remaining kind, include mainly tobacco black shank, bacterial wilt, rust, anthracnose, black lowering and white wherein nearly 10 kinds of weight of causing harm
Powder disease etc..Some cause of diseases can cause harm in each breeding time of tobacco, rhizome disease such as tobacco black shank (Phytophthora
Parasitica var.Nicotianae) and tobacco bacterial wilt (Ralstonia solanacearum), tobacco in seedling stage at
The ripe phase can cause harm, and Alternaria alternate is caused harm heavier, and the happening and prevelence with the weather of different year at later stages, is caused
Vega Severe Reduction seriously affects yield of tobacco and quality.
Tobacco bacterial wilt is also known as bacterialo wilt disease (bacterial wilt), is by Ralstonia solanacearum
(R.solanacearum) a kind of typical vascular bundle diseases, root, stem, Ye Gebu can be aggrieved caused by, catch an illness blade initial stage
Still it is that green is suspended on stem, therefore claims " bacterial wilt ".When hot weather, the blade of diseased plant is burnt in irregular, and blade is dried,
The fall off stem of diseased plant of edge is in vertical, and hang down on stem dead blade；When soil moisture is big, the soft rotten stickness of root causes complete stool
It is withered.For tobacco bacterial wilt based on Tobacco Root portion of causing harm, most significant symptom is withered, and withered speed is quickly, once hair
Disease can cause complete stool dead, be that a War Torn venereal disease is done harm on tobacco.Tobacco bacterial wilt is that China's cigarette district causes harm most heavy one kind
Tobacco bacteriosis, only Jilin and Heilungkiang there is no this disease occur, at present China Yangtze river basin and on the south cigarette district generally occur,
Had the tendency that north developing in recent years, Shandong, Henan and Liaoning part cigarette district also have generation, some areas to cause harm heavier.
Tobacco black shank is the destructive soil-borne disease caused by Phytophthora nicotianae Breda (P.parasitica), incidence
Height has a very wide distribution, caused by economic loss it is huge.Nineteen fifty, the disease was found for the first time in Chinese the Yellow River and Huai He River cigarette district, removed northeast at present
Outside cigarette district sporadicly occurs, each cigarette district in the whole nation has generation, the production for tobacco of seriously causing harm.In recent years, due to China's continuous cropping vega
Area expands year by year, and the continuous cropping time limit is continuously increased, and has aggravated the prevalence of tobacco black shank.
Alternaria alternate is the tikka in the tobacco leaf maturity period caused by alternaria nees fungus Alternaria alternata
Class disease, each cigarette district in China have generation.In recent years, the Yellow River and Huai He River, northeast cigarette district local vega, it has also become destructive disease it
One, it is not only caused, and tobacco leaf is incomplete, grade declines, but also since interior quality is uncoordinated, makes jealous variation, reduce work
Industry use value.
Currently, the study on prevention of tobacco bacterial wilt, balck shank and rust, including from breeding for disease resistance, cultural control, change
It learns prevention and biological control etc. and has carried out a large amount of research work, but since cultivated land resource is insufficient, of high cost, high-quality anti-
The restraining factors such as sick kind shortage, so far still without ideal control measure.And Biocontrol microorganism be in crop green prevention and control most
One of means of potentiality, that has reported so far there is the active microorganism of biological and ecological methods to prevent plant disease, pests, and erosion to be mainly derived from host's plant Ralstonia solanacearum
The rhizosphere soil of object and the endogenous cycle border of root, including it is pseudomonad (Pseudomonas), bacillus (Bacillus), short
Bacillus (Brevibacillus), ground bacillus (Geobacillus), relies series bacillus (Paenibacillus)
Propylhomoserin bacillus (Lysinibacillus), Stenotrophomonas (Stenotrophomonas), acinetobacter
(Acinetobacter), Enterobacter (Enterobacter), streptomycete (Streptomyces), arthrobacterium
(Arthrobacter), frankia (Frankia), aspergillus (Aspergillus), trichoderma (Trichoderma), mould
(Penicillium) and bacteriophage etc., ironically, some pseudomonads for being isolated from acid soil environment also show
Very high antagonistic activity.It is much ground although the biological control about the fungies such as tobacco bacterial wilt or bacteriosis both at home and abroad has had
Study carefully, and achieve certain effect, but the acid resistance of these bacterial strains is poor, applies plate caused by acid soil, especially continuous cropping
Tie, be acidified in serious soil and cannot give full play to its antagonism, and most biocontrol bacterial strains only for one type fungi or
Bacteriosis has preferable control effect, and the defence that Biocontrol Effect is all had with acid resistance and to soil-borne disease pathogen is false
There has been no reports for unit cell bacterial strain.
Invention content
For the above-mentioned problems in the prior art, the purpose of the present invention is to provide one plant of acid resistance disease prevention growth-promotings to give birth to
Defence pseudomonad strain is suitable for the biological control of tobacco bacterium, fungal disease under the conditions of acid soil, is that tobacco is main
The preventions such as rhizome disease bacterial wilt, balck shank and rust provide new microbial resources.
To achieve the above object, the present invention relates to following technical schemes：
The first aspect of the invention provides one plant of acid resistance disease prevention growth-promoting defence pseudomonad (Pseudomonas
Protegens) CLP-6, it is general which has been preserved in China Committee for Culture Collection of Microorganisms on October 27th, 2016
Logical microorganism center (address：City of BeiJing, China Chaoyang District North Star West Road 1 institute 3), biological deposits number are CGMCC
No.13205。
The second aspect of the invention provides above-mentioned defence pseudomonad (Pseudomonas protegens) CLP-6's
Cultural method, including pseudomonad (Pseudomonas protegens) CLP-6 will be defended and be placed in NB fluid nutrient mediums and train
It supports, condition of culture is aerobic culture at 30 DEG C, such as shaking table (30 DEG C, 150rpm) shaken cultivation.
Wherein, the NB culture medium prescriptions (g/L) are：Beef extract 3.0；Peptone 5.0；Glucose 2.5；pH7.0±
0.2；
The third aspect of the invention, provides a kind of microbial bacterial agent, and the microbial bacterial agent includes that above-mentioned defence is false single
Born of the same parents bacterium (Pseudomonas protegens) CLP-6's or defence pseudomonad (Pseudomonas protegens) CLP-6
Culture.The microbial bacterial agent, which has, inhibits tobacco black shank bacterium, ralstonia solanacearum and brown spot pathogen and promotion tobacco growing
Effect.
Preferably, the dosage form of microbial bacterial agent is that wettable powder, water dispersible granules, aqueous suspension agent or dispersible oil are outstanding
Floating agent；
Preferably, further include acceptable auxiliary material in Pesticide Science in microbial bacterial agent, it is acceptable in the Pesticide Science
One kind in dispersant, wetting agent, disintegrant, binder, antifoaming agent, antifreeze, thickener, filler and solvent of auxiliary material or
It is a variety of.The present invention is not particularly limited the source etc. of acceptable auxiliary material in the Pesticide Science, is generally using commercial product
It can.
The fourth aspect of the invention, provides a kind of microbial-bacterial fertilizer, and the microbial-bacterial fertilizer includes that above-mentioned defence is false single
Born of the same parents bacterium (Pseudomonas protegens) CLP-6's or defence pseudomonad (Pseudomonas protegens) CLP-6
Culture.The microbial-bacterial fertilizer, which has, inhibits tobacco black shank bacterium, ralstonia solanacearum and brown spot pathogen and promotion tobacco growing
Effect.
Preferably, microbial-bacterial fertilizer also contains organic matter, full potassium, full nitrogen, for providing nutrient.
The fifth aspect of the invention provides above-mentioned defence pseudomonad (Pseudomonas protegens) CLP-
6, the application of microbial bacterial agent or microbial-bacterial fertilizer in tobacco bacterium, fungal disease prevention.
Preferably, the tobacco bacterium, fungal disease include but not limited to tobacco bacterial wilt, tobacco black shank and tobacco
Rust.
The sixth aspect of the invention, provide above-mentioned defence pseudomonad (Pseudomonas protegens) CLP-6,
The application of microbial bacterial agent or microbial-bacterial fertilizer in promoting tobacco growing.
Preferably, the bacterial strain, microbial bacterial agent or microbial-bacterial fertilizer are more suitable for playing a role in acid condition；
Further, the acid condition pH is 5.5-6.5.
Beneficial effects of the present invention：
The present invention is mixed from tobacco bacterial wilt and balck shank for the first time by indoor flat plate face-off method and greenhouse pot culture control effect
The tobacco healthy tree Rhizosphere sampling of hair field filters out one plant of defence pseudomonad (Pseudomonas protegens) CLP-6, institute
Stating bacterial strain has acid resistance, and diseases prevention, growth-promoting effect are all good, and its resistance is strong, which has both pathogen antagonistic activity, and right
The acidity tolerance range of soil is wide, antimicrobial spectrum is wide, has certain answer in the prevention of tobacco bacterial wilt, balck shank and rust
Use potentiality to be exploited.Meanwhile under the increasingly severe overall background of China's acidification situation, the research of the bacterial strain is for various soil properties
Under control of plant disease also there is certain reference, be tobacco bacterial wilt in acid soil, balck shank and rust
Green prevention and control provide microbial resources, excavate microbial secondary metabolites so that improve acid soil in bacterial wilt, balck shank and
Rust biocontrol effect establishes important foundation.
Description of the drawings
Fig. 1 is CLP-6 flat-plate bacterial colonies, morphological features figure.
Fig. 2 is CLP-6 degermings supernatant to the inhibiting effect of ralstonia solanacearum, and wherein left figure is the diffusion examination of Oxford cup tablet
It tests, right figure is CLP-6 treated ralstonia solanacearum figures.
Fig. 3 is CLP-6 under Electronic Speculum to the extent of the destruction figure of Ralstonia solanacearum thalline.
Fig. 4 is the inhibiting effect lithograph that CLP-6 degermings supernatant grows black shank bacterium mycelia, wherein a left side is CLP-6
Black shank bacterium deformity mycelia after processing, the right side are the normal mycelia of black shank bacterium.
Fig. 5 is the inhibiting effect light microscopic figure that CLP-6 degermings supernatant grows black shank bacterium mycelia, wherein a left side is CLP-6
Black shank bacterium deformity mycelia after processing, the right side are the normal mycelia of black shank bacterium.
Fig. 6 is antagonistic activity of the CLP-6 thalline to brown spot pathogen.
Fig. 7 is the influence light microscopic figure that CLP-6 thalline grow brown spot pathogen mycelia, wherein the right side is red star after CLP-6 processing
Germ deformity mycelia, a left side are the normal mycelia of brown spot pathogen.
Fig. 8 is the phosphate solubilization figure of CLP-6 degerming supernatants.
Fig. 9 is the ability of dissolving potassium figure of CLP-6 degerming supernatants.
Figure 10 is production protease ability (under acid condition) figure of CLP-6 degerming supernatants.
Specific implementation mode
It is noted that following detailed description is all illustrative, it is intended to provide further instruction to the application.Unless another
It indicates, all technical and scientific terms used herein has usual with the application person of an ordinary skill in the technical field
The identical meanings of understanding.
It should be noted that term used herein above is merely to describe specific implementation mode, and be not intended to restricted root
According to the illustrative embodiments of the application.As used herein, unless the context clearly indicates otherwise, otherwise singulative
It is also intended to include plural form, additionally, it should be understood that, when in the present specification using term "comprising" and/or " packet
Include " when, indicate existing characteristics, step, operation, device, component and/or combination thereof.
In conjunction with specific example, the present invention is further illustrated, and following instance is not right merely to the explanation present invention
Its content is defined.If the experiment actual conditions being not specified in embodiment, usually according to normal condition, or according to reagent public affairs
The recommended condition of department；Reagent as used in the following examples, consumptive material etc., are commercially available unless otherwise specified.
In the specific embodiment of the present invention, one plant of acid resistance disease prevention growth-promoting defence pseudomonad is provided
(Pseudomonas protegens) CLP-6, the bacterial strain are preserved in Chinese microorganism strain preservation on October 27th, 2016
Administration committee's common micro-organisms center (address：City of BeiJing, China Chaoyang District North Star West Road 1 institute 3), biological deposits number
For CGMCC No.13205.
In the present invention, by measuring bacterial strain 16S rDNA gene orders (see SEQ ID NO.1), gyrB gene orders
(see SEQ ID NO.2), in conjunction with bacterium colony, morphological features (see Fig. 1), physiological and biochemical property (see the table below), and final determine should
Bacterial strain belongs to defence pseudomonad (Pseudomonas protegens).
Table bacterial strain CLP-6 physiological and biochemical properties
Note：+:Positive ,-Negative, W：Weak Positive
In the still another embodiment of the present invention, above-mentioned defence pseudomonad (Pseudomonas is provided
Protegens) the cultural method of CLP-6, including pseudomonad (Pseudomonas protegens) CLP-6 will be defended and be placed in
It is cultivated in NB fluid nutrient mediums, condition of culture is aerobic culture at 30 DEG C, such as shaking table (30 DEG C, 150rpm) shaken cultivation.
Wherein, the NB culture medium prescriptions (g/L) are：Beef extract 3.0；Peptone 5.0；Glucose 2.5；pH7.0±
0.2；
In the still another embodiment of the present invention, a kind of microbial bacterial agent is provided, the microbial bacterial agent includes above-mentioned
Defend pseudomonad (Pseudomonas protegens) CLP-6 or defence pseudomonad (Pseudomonas protegens)
The culture of CLP-6.The microbial bacterial agent have inhibit tobacco brown spot pathogen, black shank bacterium and with ralstonia solanacearum and promotion
The effect of tobacco growing.
In the still another embodiment of the present invention, the dosage form of microbial bacterial agent is wettable powder, water dispersible granules, water
Suspending agent or dispersible oil-suspending agent；
Preferably, further include acceptable auxiliary material in Pesticide Science in microbial bacterial agent, it is acceptable in the Pesticide Science
One kind in dispersant, wetting agent, disintegrant, binder, antifoaming agent, antifreeze, thickener, filler and solvent of auxiliary material or
It is a variety of.The present invention is not particularly limited the source etc. of acceptable auxiliary material in the Pesticide Science, is generally using commercial product
It can.
Wherein, the dispersant is anionic dispersing agent and/or non-ionic dispersing agent, can be selected from lignin sulfonic acid
Sodium, naphthalenesulfonic acid-formaldehyde condensate, sodium methylene bis-naphthalene sulfonate, formaldehyde condensation products sulfate, polycarboxylate, alkyl phenol polyoxy second
It is one or more in alkenyl phosphate and polyoxyethylene carboxylate；
The wetting agent can be selected from lauryl sodium sulfate, neopelex, spaonin powder, spaonin powder, tea seed cake
It is one or more in powder and Nekal BX；
The disintegrant can be selected from one kind or more in bentonite, ammonium sulfate, aluminium chloride, urea, magnesium chloride and glucose
Kind；
The binder can be selected from starch, diatomite, cyclodextrin, rosin, carboxymethyl cellulose, carboxyethyl cellulose and carboxylic
It is one or more in methylcellulose salt；
The antifoaming agent can be selected from C8~C20 aliphatic alcohols compound, C10~C20 saturated fat acids compound, epoxy
It is one or more in soybean oil, ethyl alcohol, silicone compound and organic silicone oil；
The antifreeze can be selected from sorbierite, ethylene glycol, polyethylene glycol, propylene glycol, glycerine, urea and sodium chloride
It is one or more；
The thickener can be selected from one or more in gelatin, xanthans, polyethylene glycol and polyvinyl alcohol；
The filler can be selected from one kind or more in precipitated calcium carbonate, diatomite, bentonite, attapulgite and white carbon
Kind；
The solvent can be selected from water (preferably deionized water) or methyl oleate；
The still another embodiment of the present invention, provides a kind of microbial-bacterial fertilizer, and the microbial-bacterial fertilizer includes above-mentioned anti-
Defend pseudomonad (Pseudomonas protegens) CLP-6 or defence pseudomonad (Pseudomonas protegens)
The culture of CLP-6.The microbial-bacterial fertilizer, which has, inhibits tobacco brown spot pathogen, black shank bacterium and ralstonia solanacearum and promotion cigarette
The effect of grass growth.
In the still another embodiment of the present invention, the microbial-bacterial fertilizer also contains organic matter, full potassium, full nitrogen, is used for
Nutrient is provided.
In the still another embodiment of the present invention, above-mentioned defence pseudomonad (Pseudomonas is provided
Protegens) the application of CLP-6, microbial bacterial agent or microbial-bacterial fertilizer in tobacco bacterium, fungal disease prevention.
In the still another embodiment of the present invention, the tobacco bacterium, fungal disease include but not limited to that tobacco blueness is withered
Disease, tobacco black shank and Alternaria alternate.
In the still another embodiment of the present invention, above-mentioned defence pseudomonad (Pseudomonas is provided
Protegens) the application of CLP-6, microbial bacterial agent or microbial-bacterial fertilizer in promoting tobacco growing.
In the still another embodiment of the present invention, the bacterial strain, microbial bacterial agent or microbial-bacterial fertilizer are more suitable for acid
It plays a role in property condition；
In the still another embodiment of the present invention, the acid condition pH is 5.5-6.5.
Explanation is further explained to the present invention by the following examples, but is not construed as limiting the invention.
The antagonistic activity verification of embodiment 1CLP-6 and its fermentation broth on tobacco bacterial wilt, balck shank germ：To from Hunan
The Zhangjiajie City Cili County towns Jiang Ya cigarette strain diseased plant antagonistic activity is verified.By NA culture mediums and appropriate acidic buffer
(sodium acetate of 0.2mol/L and the acetic acid of 0.3mol/L are according to volume ratio 1：9 mixing) mixing, be prepared into pH be respectively 5.5,
6.0, the NA culture mediums of 6.5,7.0 and 7.5 different pH values measure CLP-6 bacterial strain fermentation liquors not using Odontothrips loti
With the difference of bacteriostatic activity under the conditions of acid-base value.CLP-6 is inoculated in 30 DEG C of culture 48h on NA tablets, then tablet is used to stand facing each other
Method is inoculated with black shank bacterium bacteria cake (a diameter of 5mm) and rust that is, on oat and potato dextrose medium (PDA) tablet
Bacterium is placed in 30 DEG C of culture 5d, the inhibition to Ralstonia solanacearum with oese picking CLP-6 in the equidistant scribing line in bacteria cake both sides
Using Oxford cup Agar diffusion test (such as Fig. 2)；CLP-6 is inoculated in beef broth peptone liquid medium (NB), 30 DEG C,
150rpm shaken cultivations 2 days take appropriate zymotic fluid and sterile supernatant (being filtered through biofilter) to be placed in the ox containing Ralstonia solanacearum
In Oxford cup on gravy peptone culture medium tablet (NA), stationary culture 2 days, suppressions of the observation CLP-6 to above-mentioned each pathogenic bacteria
Situation processed.From indoor flat plate Bacteriostatic Effect, CLP-6 thalline, zymotic fluid and sterile supernatant are to tobacco brown spot pathogen, balck shank
Bacterium and ralstonia solanacearum all have good inhibiting effect (see Fig. 2,4 and 6, table 1,2).
From table 1,2 as can be seen that in acid (5.5≤pH≤6.5) range, CLP-6 is to ralstonia solanacearum, black shank bacterium
Inhibition is stronger, and higher than neutral or alkaline condition, illustrates that acid condition is conducive to the bacterial strain and plays best fungistatic effect, because
The biological control of bacterial wilt, balck shank will have very big potentiality under the conditions of the bacterial strain is applied to acid soil by this.
Fungistatic effects of the 1 acidproof antagonistic strain CLP-6 of table to Ralstonia solanacearum
Note：Significant difference (p≤0.01) is represented with alphabetical difference after column data.
2 acidproof antagonistic strain CLP-6 bacterial strains of table antagonistic activity in acid and neutrallty condition compares
Note：Significant difference (p≤0.05) is represented with alphabetical difference after column data.
Embodiment 2CLP-6 and its zymotic fluid are to ralstonia solanacearum thalline extent of the destruction：CLP-6 is inoculated in 30 DEG C on NA tablets
48h is cultivated, tablet face-off method is then used, Oxford cup Agar diffusion test is used to the inhibition of Ralstonia solanacearum；I.e. by CLP-6
It is inoculated in beef broth peptone liquid medium (NB), 30 DEG C, 150rpm shaken cultivations 2 days, takes appropriate zymotic fluid and sterile
Supernatant (being filtered through biofilter) is placed in the Oxford cup on the beef broth peptone culture medium tablet (NA) containing Ralstonia solanacearum,
Stationary culture 2 days, inhibition situations of the observation CLP-6 to above-mentioned each pathogenic bacteria.From indoor flat plate Bacteriostatic Effect, CLP-6 bacterium
Body, zymotic fluid and sterile supernatant have good inhibiting effect to tobacco ralstonia solanacearum (see Fig. 2).
The antibacterial substance of CLP-6 strain secretes is the CLP- that will be activated to the antagonistic activity of the destruction of Ralstonia solanacearum thalline
6, which are inoculated in 30 DEG C of shaken cultivation 48h in NB culture mediums, obtains zymotic fluid, the supernatant after taking it to remove thalline and Ralstonia solanacearum bacteria suspension
(a concentration of 2 × 108Cfu/ml) isometric mixing, blank control are equivalent sterile water and bacteria suspension mixed processing.It handles for 24 hours,
The mixed liquor managed everywhere in after diluting is taken to be observed under scanning electron microscope.From the results, it was seen that the bateriostatics of CLP-6 strain secretes
Confrontation Ralstonia solanacearum thalline destruction is stronger, and rod-shaped thalline is caused to deform, and content leaks, and the Ralstonia solanacearum bar of sterile water control
Shape thalline is normal (see Fig. 3).
The inhibiting effect that embodiment 3CLP-6 and its zymotic fluid grow black shank bacterium mycelia：Using black shank bacterium as target
Bacterium is inoculated with CLP-6 in 30 DEG C of culture 48h on NA tablets, then uses tablet face-off method, i.e., be inoculated with black shin on oat tablet
Germ is placed in 30 DEG C of culture 5d, picking black shank bacterium bacterium with oese picking CLP-6 and the equidistant scribing line centered on bacteria cake
As a result cake edge mycelia microscopy under microscope is shown through CLP-6 treated black shank bacterium bacteria cake edge mycelia and normal bacterium
Silk is compared, and the cytoplasm in hyphal cell is collected as spherical, is detached from cell membrane and cell wall, therefore influences its normal growth
(see Fig. 4).
Influence of the thalline of embodiment 4CLP-6 bacterial strains to the inhibiting effect of brown spot pathogen and its to mycelia growth：
Using brown spot pathogen as target bacterium, that is, CLP-6 bacterial strains are inoculated in 30 DEG C of culture 48h on NA tablets, then use tablet
Face-off method, i.e., be inoculated with brown spot pathogen on PDA plate, and with oese picking CLP-6 bacterial strains, both sides are equidistant centered on bacteria cake
Parallel scribing is placed in 30 DEG C of culture 5d, and then picking brown spot pathogen bacteria cake edge mycelia microscopy under microscope, as a result shows warp
CLP-6 treated brown spot pathogen bacterium colony growths are suppressed, and the brown spot pathogen bacterium colony for closing on CLP-6 bacterial strains is slow-growing, bacterium
It falls and develops (Fig. 6) in sleeve configuration.And through CLP-6 treated brown spot pathogen mycelia and normal Hyphal form ratio, inhibiting effect table
It is now mycelia deformity, the deformation of mycelia compartment and swelling, mycelia content is disperseed or aggregation, and mycelia is without vitality, growth retardation
(Fig. 7).
Embodiment 5CLP-6 and its zymotic fluid have Soluble phosphorus effect：Acidproof antagonistic strain culture is measured with molybdenum antimony resistance colorimetric method
Water-soluble phosphorus content in liquid determines the bacterial strain phosphate solubilization.It takes acidproof antagonistic strain standard bacterium solution to be inoculated in pH by 1% to be respectively
In 5.5 and 7.0 Soluble phosphorus culture medium, 28 DEG C, 170r/min shaken cultivation 5d, by cultured zymotic fluid in centrifuge
4000r/min centrifuges 20min, pours out supernatant and be settled to 50mL, after precipitation adds 1g quartz sands to grind 10min in mortar
Under being washed with distillation, lapping liquid 4000r/min in centrifuge, secondary centrifuging 20min discard precipitation, and supernatant is equally fixed
It is 50mL to hold.The supernatant centrifuged twice is merged, water-soluble phosphorus yield in strains tested zymotic fluid is carried out, determines
Its phosphate solubilization.Each 3 repetitions of processing.
Supernatant 5ml is taken, 0.5mol/L NaHCO are added3Then solution 10mL is added distilled water 35mL, then uses liquid relief
Pipe draws the anti-color-developer reagent 5.0mL of molybdenum antimony, shakes up, and after being stored at room temperature 30min, colorimetric is carried out under 720nm wavelength, is adjusted not
The OD of the Soluble phosphorus culture medium of inoculation720Value is 0, measures the absorbance of strain to be tested supernatant, compares to obtain the bacterium with standard curve
Titanium pigment content in strain supernatant.
CLP-6 points after taking oese picking to activate are connected on Soluble phosphorus culture medium, and 30 DEG C of culture 48h observe CLP-6 bacterium colonies
Whether surrounding generates haloing, if there is haloing generation, is shown to be positive.The results show that CLP-6 periphery of bacterial colonies generates one fixed width
Haloing (such as Fig. 8) illustrates that the bacterial strain has certain phosphate solubilization.
Test result such as Fig. 5, shown in table 3, water-soluble phosphorus content difference is aobvious in CLP-6 culture solutions under acidity-neutrallty condition
It writes, under the acid condition of pH 5.5, water-soluble phosphorus content is 46.2mg/L；It is 21.1mg/L under the neutrallty condition of pH 7.0.
Therefore, (pH 5.5) phosphate solubilization is better than neutrallty condition (pH7.0) to CLP-6 in acid condition.
3 acidproof antagonistic strain CLP-6 of table water-soluble phosphorus contents in acid-neutral culture medium
Note：Significant difference (p≤0.05) is represented with alphabetical difference after column data.
Embodiment 6CLP-6 and its zymotic fluid have potassium decomposing effect：Acidproof antagonistic strain culture is measured using Sodium Tetraphenylborate Method
Water-soluble phosphorus content in liquid determines the bacterial strain phosphate solubilization.It takes acidproof Antagonistic Fungi standard bacterium solution to be inoculated in pH by 1% to be respectively
In 5.5 and 7.0 potassium decomposing culture medium, 28 DEG C, 170r/min shaken cultivation 5d, by cultured zymotic fluid in centrifuge
4000r/min centrifuges 20min, pours out supernatant and be settled to 50mL, after precipitation adds 1g quartz sands to grind 10min in mortar
Under being washed with distillation, lapping liquid 4000r/min in centrifuge, secondary centrifuging 20min discard precipitation, and supernatant is equally fixed
It is 50mL to hold.The supernatant centrifuged twice is merged, the measurement of water-soluble potassium in strains tested zymotic fluid is carried out, is determined
Its ability of dissolving potassium.Each 3 repetitions of processing.
It takes above-mentioned centrifugation solution 5.0mL in colorimetric cylinder, 1mL formaldehyde-EDTA screening agents is added, shakes up, is inhaled with pipette
It takes sodium tetraphenylborate solution 1.0mL to be added in colorimetric cylinder, after shaking up, 15min is stored at room temperature, after shaking up again, in 420nm wavelength items
Colorimetric is carried out under part, adjusts the OD for the potassium decomposing culture medium not being inoculated with420Value is 0, measures the absorbance of strain to be tested supernatant, with
Standard curve compares to obtain titanium pigment content in the bacterial strain supernatant.
The CLP-6 points after oese picking activationization are taken to be connected on potassium decomposing culture medium, 30 DEG C of culture 48h observe CLP-6 bacterium
Whether haloing is generated around falling, if there is haloing generation, is shown to be positive.As a result (such as Figure 10) is shown, CLP-6 periphery of bacterial colonies generates
The haloing of one fixed width illustrates that the bacterial strain has certain ability of dissolving potassium.Invalid potassium in soil can be switched to effective potassium by potassium solubilizing bacteria,
Increase the potassium element in soil.Tobacco is that happiness potassium crop provides enough potassium nutritions for cigarette strain if CLP-6 is applied to tobacco,
To have larger application value in terms of promoting quality of tobacco and yield.
Test result such as Fig. 9, shown in table 4, water-soluble potassium contains in acidproof antagonism CLP-6 culture solutions under acidity-neutrallty condition
It is not notable to measure difference, under the acid condition of pH 5.5, water-soluble potassium content is 3.2mg/L in culture solution；In pH 7.0
Property under the conditions of be 2.9mg/L.Therefore, CLP-6 ability of dissolving potassium under the conditions of (pH 5.5) and neutral (pH 7.0) in acid condition
Quite.
Table 4CLP-6 water-soluble potassium contents in acid-neutral culture medium
Note：Significant difference (p≤0.05) is represented with alphabetical difference after column data.
Embodiment 7CLP-6 and its zymotic fluid have the ability of extracellular proteinase：With reference to forint phenol reagent process and fitted
When adjustment, take and be inoculated in by 1% in the NB of pH 5.5 and 7.0 under standard bacteria suspension aseptic condition, each handle 3 repetitions,
28 DEG C, 170r/min oscillation 3d, 10000r/min centrifugation 10min, take 1mL supernatants as enzyme solution, are respectively placed in 3 test tubes simultaneously
Be placed in 37 DEG C of water-baths and preheat 2min, be then added again into test tube after the casein solutions that have been warmed up of 1mL mix well in
37 DEG C of reaction 30min, then the solution of trichloroacetic acid of addition 2ml 0.4mol/L is anti-in 37 DEG C after mixing well into each test tube
30min is answered to make reaction terminating.10000g centrifuges 10min, takes 1mL supernatants in test tube, and 5mL 0.4mol/ are added into test tube
The Na of L2CO3Solution and 1mL forint phenol reagents are developed the color 20min in 40 DEG C of water-baths after mixing.Blank control is set simultaneously, i.e., in enzyme
Solution of trichloroacetic acid is first added in liquid, casein solution is added afterwards.After colour developing light absorption value is measured under the wavelength of 680nm.
Test result such as Figure 10, table 5, are computed, in neutral-acid culture solution, acidity cultures of the CLP-6 in pH5.5
In liquid, the bacterium solution enzyme activity of CLP-6 is 49.89U/mL；It is 38.15U/mL in the neutral culture solution of pH 7.0.Therefore, acid item
Part is suitable for CLP-6 extracellular proteinases.
5 acidproof antagonistic strain CLP-6 enzymatic productivities (acid condition pH5.5) of table
Note：Significant difference (p≤0.05) is represented with alphabetical difference after column data.
Embodiment 8CLP-6 and its zymotic fluid have the ability of secretion phosphate：Phosphate in acid-neutral bacterium solution
Vitality test takes with reference to occluded corrosion cell and is inoculated in pH5.5's and 7.0 by 1% under standard bacteria suspension aseptic condition
In Pikovskaya fluid nutrient mediums, 3 repetitions, 28 DEG C, 170r/min oscillation 3d, 10000r/min centrifugations are each handled
10min takes 1mL supernatant distilled water to dilute 10 times.The accurate zymotic fluid drawn after dilution, and with nonvaccinated sky after sterilizing
White Pikovskaya fluid nutrient mediums as a contrast, measure absorbance value, the phosphorus content in test liquid are checked on standard curve.
Test result is as shown in Table 5,6, the CLP-6 phosphate vigor significant differences in acid-neutral culture solution, in pH
Under 5.5 acid condition, CLP-6 phosphate vigor is 5.12U/mL；Under the neutrallty condition of pH 7.0, enzyme activity is
8.27U/mL.Illustrate under solutions of weak acidity (pH 5.5), CLP-6 produces phosphate ability and is less than neutrallty condition (pH 7.0).
6 acidproof antagonistic strain CLP-6 of table phosphate vigor in acid-neutral culture medium
Note：Significant difference (p≤0.01) is represented with alphabetical difference after column data.
The growth-promoting functions of embodiment 9CLP-6 and its fermentation broth on tobacco：Experiment uses nutrient discs planting patterns, in nature
It is carried out in soil, the mode of seedling leaching root handles tobacco seedlings, and method is as follows：By crop field loam:Farm manure:Vermiculite is according to 7:2:1 ratio
Matrix is prepared, natural soil is filled in seedlings nursing plate.The careful tobacco seedlings for digging out 3-4 piece true leaves, gently shake off to be attached to root
Soil, and be soaked in the prepared standard biological and ecological methods to prevent plant disease, pests, and erosion bacteria suspension to be measured of pre-selection (concentration is about 108Cfu/mL in), after 40min
It takes out and transplants in seedlings nursing plate, after being spaced 7d, with standard bacteria suspension pouring root 1 time, 5mL/ plants；Control is that clear water is handled.Often handle
It 10 plants, repeats three times, in 30 DEG C of greenhouse moisturizing culture.20d after last processing, randomly selects 10 plants of tobacco seedlings, carefully by the whole strain of seedling
It digs out, washes away root soil, measure the indexs such as its plant height, whole strain fresh weight, root fresh weight.Then 180 DEG C drying to constant weight, survey whole strain
Dry weight and root dry weight.Growth-promoting test result shows and (is shown in Table 7,8), when being inoculated with the tobacco seedlings that CLP-6 is planted in soil naturally, Neng Gouming
Aobvious to promote tobacco growing, whole strain fresh weight and dry weight increase significantly, and leaf color is dark green, and blade is plump and big；In natural bacteria soil, survey
Each index for determining tobacco seedlings after CLP-6 is handled is higher than compareing, and compared with CK, the bacterial strain of CLP-6 processing can be effectively promoted tobacco
Growth, plant height, whole strain fresh weight and dry weight, root fresh weight, root dry weight development difference is notable, and the cigarette strain through CLP-6 processing is to plant height, whole
Strain fresh weight, dry weight, root dry weight growth-promoting are with obvious effects, increase by 31.2%, 40.0%, 70.6%, 45.2% respectively, this bacterial strain growth-promoting
Effect is extremely strong, while illustrating that CLP-6 bacterium are safe to tobacco.
Acidproof antagonistic strain CLP-6 growth-promoting functions in table 7pH5.5 soil
Note：Significant difference (p≤0.05) is represented with alphabetical difference after column data.
Acidproof antagonistic strain CLP-6 growth-promoting functions in table 8 normal pH (7.1) soil
Note：Significant difference (p≤0.05) is represented with alphabetical difference after column data.
In conclusion bacterial strain defence pseudomonad (Pseudomonas protegens) CLP-6 provided by the invention is abundant
The type of biological and ecological methods to prevent plant disease, pests, and erosion pseudomonad and its scope is applied, which in different medium shows strong growth ability and steady
Fixed activity has the value further researched and developed.
The present invention filters out one plant of defence pseudomonad for the first time by indoor flat plate culture and live body greenhouse preventive effect
(Pseudomonas protegens) CLP-6, the strain isolation is from acid tobacco root soil, to tobacco bacterial wilt, black shin
The pathogenic strain of disease and rust all has antagonistic activity, while having acid resistance, Soluble phosphorus, potassium decomposing, extracellular proteinase, phosphate
The abilities such as enzyme.The CLP-6 and its microbial bacterial agent can effectively prevent soil-borne disease of tobacco under the conditions of continuous cropping or acid soil,
It is the microbial pesticide resource of novel high-quality, has broad application prospects.
It should be noted that above example is only used to illustrate the technical scheme of the present invention rather than is limited.Although ginseng
It is described the invention in detail according to given example, but those skilled in the art can be as needed to this hair
Bright technical solution is modified or replaced equivalently, without departing from the spirit of the technical scheme of the invention and range.
SEQUENCE LISTING
<110>Tobacco Institute, Chinese Academy of Agricultural Science
<120>One plant of acid proof defence pseudomonad CLP-6 and its application
<130>
<160> 1
<170> PatentIn version 3.5
<210> 1
<211> 1381
<212> DNA
<213>Defend pseudomonad Pseudomonas protegens 16s rDNA
<400> 1
cggcagcacg ggtacttgta cctggtggcg agcggcggac gggtgagtaa tgcctaggaa 60
tctgcctagt agtgggggat aacgtccgga aacgggcgct aataccgcat acgtcctacg 120
ggagaaagtg ggggatcttc ggacctcacg ctattagatg agcctaggtc ggattagcta 180
gttggtgagg taatggctca ccaaggcgac gatccgtaac tggtctgaga ggatgatcag 240
tcacactgga actgagacac ggtccagact cctacgggag gcagcagtgg ggaatattgg 300
acaatgggcg aaagcctgat ccagccatgc cgcgtgtgtg aagaaggtct tcggattgta 360
aagcacttta agttgggagg aagggcagtt acctaatacg tgattgtttt gacgttaccg 420
acagaataag caccggctaa ctctgtgcca gcagccgcgg taatacagag ggtgcaagcg 480
ttaatcggaa ttactgggcg taaagcgcgc gtaggtggtt tgttaagttg gatgtgaaag 540
ccccgggctc aacctgggaa ctgcatccaa aactggcaag ctagagtatg gtagagggtg 600
gtggaatttc ctgtgtagcg gtgaaatgcg tagatatagg aaggaacacc agtggcgaag 660
gcgaccacct ggactgatac tgacactgag gtgcgaaagc gtggggagca aacaggatta 720
gataccctgg tagtccacgc cgtaaacgat gtcaactagc cgttgggagc cttgagctct 780
tagtggcgca gctaacgcat taagttgacc gcctggggag tacggccgca aggttaaaac 840
tcaaatgaat tgacgggggc ccgcacaagc ggtggagcat gtggtttaat tcgaagcaac 900
gcgaagaacc ttaccaggcc ttgacatcca atgaactttc tagagataga ttggtgcctt 960
cgggaacatt gagacaggtg ctgcatggct gtcgtcagct cgtgtcgtga gatgttgggt 1020
taagtcccgt aacgagcgca acccttgtcc ttagttacca gcacgttatg gtgggcactc 1080
taaggagact gccggtgaca aaccggagga aggtggggat gacgtcaagt catcatggcc 1140
cttacggcct gggctacaca cgtgctacaa tggtcggtac aaagggttgc caagccgcga 1200
ggtggagcta atcccataaa accgatcgta gtccggatcg cagtctgcaa ctcgactgcg 1260
tgaagtcgga atcgctagta atcgcgaatc agaatgtcgc ggtgaatacg ttcccgggcc 1320
ttgtacacac cgcccgtcac accatgggag tgggttgcac cagaagtagc tagtctaacc 1380
t 1381
<210> 2
<211> 652
<212> DNA
<213>Defend pseudomonad Pseudomonas protegens gyrB
<400> 2
ggtagtgaac gccctgtccg aggagctgat cctcaccgtg cgccgtagcg gcaagatctg 60
ggaacagacc tatgtccacg gtgttccgca agagcggatg aaaatcgttg gtgacagcga 120
aaccaccggt acccagatcc acttcaagcc ttcggctgaa accttcaaga acatccactt 180
cagctgggac gtcctggcca agcggatccg tgaactgtcc ttcctcaact ccggtgtcgg 240
catcgtcctc aaggacgagc gcagcggcaa ggaagagctg ttcaagtacg aaggcggcct 300
gcgggcattc gttgaatacc tgaacaccaa caagactgcg gtcaaccagg tgttccactt 360
caacatccag cgtgaagacg gcatcggcgt ggaaatcgcc ttgcagtgga acgacagctt 420
caacgagaac ctgttgtgct tcaccaacaa cattcctcag cgcgacggcg gtacccacct 480
ggtgggcttc cgttcggccc tgacccgtaa cctgaacaac tacatcgagc aggaaggcct 540
ggccaagaaa cacaaggtcg ccaccactgg tgacgacgct cgtgaaggcc tgaccgcgat 600
catctcggtg aaggtgccgg atccgaagtt cagctcccag accaaggaca ag 652
Claims (10)
1. one plant of acid resistance disease prevention growth-promoting defends pseudomonad (Pseudomonas protegens) CLP-6, the bacterial strain in
On October 27th, 2016 is preserved in China Committee for Culture Collection of Microorganisms's common micro-organisms center, and biological deposits number are
CGMCC No.13205。
2. defending the cultural method of pseudomonad (Pseudomonas protegens) CLP-6, feature described in claim 1
It is, defence pseudomonad (Pseudomonas protegens) CLP-6 is placed in NB fluid nutrient mediums and is cultivated, it is preferred that
Condition of culture is aerobic culture at 30 DEG C.
3. a kind of microbial bacterial agent, which is characterized in that the microbial bacterial agent includes defending pseudomonad described in claim 1
(Pseudomonas protegens) CLP-6 or the culture for defending pseudomonad (Pseudomonas protegens) CLP-6
Object.
4. a kind of microbial bacterial agent as claimed in claim 3, which is characterized in that the microbial bacterial agent dosage form is wettable powder
Agent, water dispersible granules, aqueous suspension agent or dispersible oil-suspending agent.
5. a kind of microbial-bacterial fertilizer, which is characterized in that the microbial-bacterial fertilizer includes above-mentioned defence pseudomonad
(Pseudomonas protegens) CLP-6 or the culture for defending pseudomonad (Pseudomonas protegens) CLP-6
Object.
6. microbial-bacterial fertilizer as claimed in claim 5, which is characterized in that the microbial-bacterial fertilizer also contains organic matter, full potassium, complete
Nitrogen.
7. being defended described in claim 1 described in pseudomonad (Pseudomonas protegens) CLP-6, claim 3 or 4
Application of microbial bacterial agent, claim 5 or 6 microbial-bacterial fertilizer in tobacco bacterium, fungal disease prevention.
8. application as claimed in claim 7, which is characterized in that the tobacco bacterium, fungal disease include tobacco bacterial wilt, tobacco
Balck shank and Alternaria alternate.
9. being defended described in claim 1 described in pseudomonad (Pseudomonas protegens) CLP-6, claim 3 or 4
Application of microbial bacterial agent, claim 5 or 6 microbial-bacterial fertilizer in promoting tobacco growing.
10. being applied as described in claim any one of 7-9, which is characterized in that the application environment is acid condition；Preferably,
The acid condition pH is 5.5-6.5.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201810438730.8A CN108570433B (en) | 2018-05-09 | 2018-05-09 | Acid-resistant pseudomonas defensins C L P-6 and application thereof |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201810438730.8A CN108570433B (en) | 2018-05-09 | 2018-05-09 | Acid-resistant pseudomonas defensins C L P-6 and application thereof |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN108570433A true CN108570433A (en) | 2018-09-25 |
| CN108570433B CN108570433B (en) | 2020-07-31 |
Family
ID=63572322
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201810438730.8A Active CN108570433B (en) | 2018-05-09 | 2018-05-09 | Acid-resistant pseudomonas defensins C L P-6 and application thereof |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN108570433B (en) |
Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN109136154A (en) * | 2018-09-29 | 2019-01-04 | 河南省农业科学院植物保护研究所 | One plant can prevent and treat tobacco black shank and root black rot and Granada pseudomonad and its application with growth-promoting functions simultaneously |
| CN109182213A (en) * | 2018-10-16 | 2019-01-11 | 云南省烟草公司普洱市公司 | A kind of tobacco-growing soil Sphingol single-cell X1-8 and its application in tobacco black shank prevention and treatment |
| CN110643538A (en) * | 2019-10-15 | 2020-01-03 | 浙江大学 | Pseudomonas fluorescens XY2F4 and application thereof in preventing and treating crop verticillium wilt |
| CN111778183A (en) * | 2020-06-30 | 2020-10-16 | 中国农业科学院烟草研究所 | Acidophilic nitrogen-producing pseudomonas strain and application thereof |
| CN112175869A (en) * | 2020-09-29 | 2021-01-05 | 烟台市林业科学研究所 | Pseudomonas and application thereof |
Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1347981A (en) * | 2000-10-11 | 2002-05-08 | 常州兰陵制药有限公司 | Pseudomonas stutzeri Zh9944 and its screening method and bacterial wilt preventing and controlling microbial prepn |
| CN105543133A (en) * | 2015-12-31 | 2016-05-04 | 山东农业大学 | Separation and application of pseudomonas with significant antagonism to plant pathogenic bacteria |
| CN106916764A (en) * | 2017-02-15 | 2017-07-04 | 中国农业科学院烟草研究所 | One plant of acid proof South Korea pseudomonad CLP 7 and its application |
| CN106978373A (en) * | 2017-05-09 | 2017-07-25 | 福建农林大学 | One plant is used for pseudomonad and the application that bacteriosis is prevented and treated |
-
2018
- 2018-05-09 CN CN201810438730.8A patent/CN108570433B/en active Active
Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1347981A (en) * | 2000-10-11 | 2002-05-08 | 常州兰陵制药有限公司 | Pseudomonas stutzeri Zh9944 and its screening method and bacterial wilt preventing and controlling microbial prepn |
| CN105543133A (en) * | 2015-12-31 | 2016-05-04 | 山东农业大学 | Separation and application of pseudomonas with significant antagonism to plant pathogenic bacteria |
| CN106916764A (en) * | 2017-02-15 | 2017-07-04 | 中国农业科学院烟草研究所 | One plant of acid proof South Korea pseudomonad CLP 7 and its application |
| CN106978373A (en) * | 2017-05-09 | 2017-07-25 | 福建农林大学 | One plant is used for pseudomonad and the application that bacteriosis is prevented and treated |
Cited By (9)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN109136154A (en) * | 2018-09-29 | 2019-01-04 | 河南省农业科学院植物保护研究所 | One plant can prevent and treat tobacco black shank and root black rot and Granada pseudomonad and its application with growth-promoting functions simultaneously |
| CN109136154B (en) * | 2018-09-29 | 2021-06-04 | 河南省农业科学院植物保护研究所 | Pseudomonas glanadensis capable of preventing and treating tobacco black shank and root black rot simultaneously and having growth promoting effect and application thereof |
| CN109182213A (en) * | 2018-10-16 | 2019-01-11 | 云南省烟草公司普洱市公司 | A kind of tobacco-growing soil Sphingol single-cell X1-8 and its application in tobacco black shank prevention and treatment |
| CN109182213B (en) * | 2018-10-16 | 2021-07-09 | 云南省烟草公司普洱市公司 | Sphingomonas X1-8 in tobacco planting soil and application thereof in prevention and treatment of tobacco black shank |
| CN110643538A (en) * | 2019-10-15 | 2020-01-03 | 浙江大学 | Pseudomonas fluorescens XY2F4 and application thereof in preventing and treating crop verticillium wilt |
| CN110643538B (en) * | 2019-10-15 | 2021-04-27 | 浙江大学 | Pseudomonas fluorescens XY2F4 and application thereof in preventing and treating crop verticillium wilt |
| CN111778183A (en) * | 2020-06-30 | 2020-10-16 | 中国农业科学院烟草研究所 | Acidophilic nitrogen-producing pseudomonas strain and application thereof |
| CN111778183B (en) * | 2020-06-30 | 2021-11-30 | 中国农业科学院烟草研究所 | Acidophilic nitrogen-producing pseudomonas strain and application thereof |
| CN112175869A (en) * | 2020-09-29 | 2021-01-05 | 烟台市林业科学研究所 | Pseudomonas and application thereof |
Also Published As
| Publication number | Publication date |
|---|---|
| CN108570433B (en) | 2020-07-31 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN105296381B (en) | One bacillus subtilis CYY-25 and its application | |
| CN108570433A (en) | One plant of acid proof defence pseudomonad CLP-6 and its application | |
| CN112266881B (en) | Bacillus amyloliquefaciens and application thereof in preventing and treating apple continuous cropping obstacle | |
| CN106916764B (en) | One plant of acid proof South Korea pseudomonad CLP-7 and its application | |
| CN101864378B (en) | Streptomyces microflavus | |
| CN109355228A (en) | One plant of forest rhizosphere bacteria intends bacillus mycoides JYZ-SD5 and its application | |
| CN101851597B (en) | Streptomyces griseoflavus for resisting alfalfa diseases and screening method thereof | |
| CN102329756B (en) | Streptomyces albospinus strain BWL15-4 for preventing and treating banana vascular wilt and application thereof | |
| CN102329755B (en) | Nonabsorbent streptomyces ahygroscopicus strain BWL58 for preventing and treating banana vascular wilt and application thereof | |
| CN103642734B (en) | Microbacterium maritypicum and application thereof in preventing sugar beet disease-causing organisms | |
| CN102168042A (en) | Pseudomonas aeruginosa and application thereof | |
| CN104388335A (en) | Bacillus amyloliquefacien FS6 and application thereof | |
| CN108384734B (en) | Biocontrol microbial inoculum for preventing and treating tobacco root and stem diseases and preparation method thereof | |
| CN103436465B (en) | Alcaligenes faecalis strain | |
| CN110079484A (en) | A kind of bacillus subtilis and its application in agricultural production | |
| Dua et al. | Effectiveness of rhizosphere bacteria for control of root rot disease and improving plant growth of wheat (Triticum aestivum L.) | |
| CN114164137B (en) | Streptomyces diastochromogenes for resisting banana vascular wilt and application thereof | |
| CN103146609B (en) | Pseudomonas fluorescens and method for preventing phytophthora capsici thereby | |
| CN104651286B (en) | A kind of soybean nodulation endophyte DEnt1302 and its preparation method and application, microbial inoculum and preparation method thereof | |
| CN110527647A (en) | A kind of bacillus and its screening technique and application from the inducible system resistant of tomato rhizosphere | |
| CN109112072B (en) | Application of trichoderma in promoting plant growth | |
| CN111334458B (en) | Biocontrol actinomycetes and application thereof in prevention and control of ginger stem basal rot or soybean epidemic disease | |
| CN109321500A (en) | One bacillus amyloliquefaciens bacterial strain and its application in prevention and treatment Oil Tea Anthracnose evil | |
| CN102533564B (en) | A kind of screening method of bio-control trichoderma in corn seedling stage root rot period | |
| CN102021131A (en) | Bacillus licheniformis strain and application thereof |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| GR01 | Patent grant | ||
| GR01 | Patent grant | ||
| TR01 | Transfer of patent right | ||
| TR01 | Transfer of patent right |
Effective date of registration: 20240131 Address after: 274300 East Middle Section of Binhe North Road, Shan County Chemical Industry Park, Xieji Town, Shan County, Heze City, Shandong Province Patentee after: Shandong Shibang Agrochemical Co.,Ltd. Country or region after: China Address before: 266101 Shandong Province, Qingdao city Laoshan District Four Keyuan Road No. 11 Patentee before: TOBACCO Research Institute CHINESE ACADEMY OF AGRICULTURAL SCIENCES Country or region before: China |