CN118086105A - Lysobacter antibioticus LMT16 and application thereof - Google Patents
Lysobacter antibioticus LMT16 and application thereof Download PDFInfo
- Publication number
- CN118086105A CN118086105A CN202410106440.9A CN202410106440A CN118086105A CN 118086105 A CN118086105 A CN 118086105A CN 202410106440 A CN202410106440 A CN 202410106440A CN 118086105 A CN118086105 A CN 118086105A
- Authority
- CN
- China
- Prior art keywords
- lmt16
- blueberry
- lysobacter
- antibiotic
- root cancer
- 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.)
- Pending
Links
- 241001660189 Lysobacter antibioticus Species 0.000 title claims description 38
- 240000000851 Vaccinium corymbosum Species 0.000 claims abstract description 129
- 235000003095 Vaccinium corymbosum Nutrition 0.000 claims abstract description 114
- 235000017537 Vaccinium myrtillus Nutrition 0.000 claims abstract description 114
- 235000021014 blueberries Nutrition 0.000 claims abstract description 114
- 206010028980 Neoplasm Diseases 0.000 claims abstract description 88
- 201000011510 cancer Diseases 0.000 claims abstract description 82
- 241000863031 Lysobacter Species 0.000 claims abstract description 36
- 230000003115 biocidal effect Effects 0.000 claims abstract description 32
- 238000011282 treatment Methods 0.000 claims abstract description 30
- 230000002265 prevention Effects 0.000 claims abstract description 21
- 230000002262 irrigation Effects 0.000 claims abstract description 14
- 238000003973 irrigation Methods 0.000 claims abstract description 14
- 238000004321 preservation Methods 0.000 claims abstract description 5
- 241000894006 Bacteria Species 0.000 claims description 36
- 241000196324 Embryophyta Species 0.000 claims description 34
- 238000000034 method Methods 0.000 claims description 31
- 238000000855 fermentation Methods 0.000 claims description 24
- 230000004151 fermentation Effects 0.000 claims description 24
- 239000001963 growth medium Substances 0.000 claims description 23
- 239000007788 liquid Substances 0.000 claims description 21
- CSNNHWWHGAXBCP-UHFFFAOYSA-L Magnesium sulfate Chemical compound [Mg+2].[O-][S+2]([O-])([O-])[O-] CSNNHWWHGAXBCP-UHFFFAOYSA-L 0.000 claims description 18
- 230000000443 biocontrol Effects 0.000 claims description 15
- 238000009630 liquid culture Methods 0.000 claims description 15
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Chemical compound O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 14
- 238000012258 culturing Methods 0.000 claims description 13
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 claims description 12
- 239000002609 medium Substances 0.000 claims description 10
- ZPWVASYFFYYZEW-UHFFFAOYSA-L dipotassium hydrogen phosphate Chemical compound [K+].[K+].OP([O-])([O-])=O ZPWVASYFFYYZEW-UHFFFAOYSA-L 0.000 claims description 9
- 239000012153 distilled water Substances 0.000 claims description 9
- 229910052943 magnesium sulfate Inorganic materials 0.000 claims description 9
- 235000019341 magnesium sulphate Nutrition 0.000 claims description 9
- FBPFZTCFMRRESA-KVTDHHQDSA-N D-Mannitol Chemical compound OC[C@@H](O)[C@@H](O)[C@H](O)[C@H](O)CO FBPFZTCFMRRESA-KVTDHHQDSA-N 0.000 claims description 7
- 229930195725 Mannitol Natural products 0.000 claims description 7
- 239000000594 mannitol Substances 0.000 claims description 7
- 235000010355 mannitol Nutrition 0.000 claims description 7
- 239000007787 solid Substances 0.000 claims description 7
- 229920001817 Agar Polymers 0.000 claims description 6
- 239000008272 agar Substances 0.000 claims description 6
- 235000015278 beef Nutrition 0.000 claims description 6
- 229940041514 candida albicans extract Drugs 0.000 claims description 6
- 239000000284 extract Substances 0.000 claims description 6
- 238000009472 formulation Methods 0.000 claims description 6
- 239000000203 mixture Substances 0.000 claims description 6
- 239000000843 powder Substances 0.000 claims description 6
- 239000011780 sodium chloride Substances 0.000 claims description 6
- 239000012138 yeast extract Substances 0.000 claims description 6
- 241000193830 Bacillus <bacterium> Species 0.000 claims description 5
- 239000001888 Peptone Substances 0.000 claims description 3
- 108010080698 Peptones Proteins 0.000 claims description 3
- 229930006000 Sucrose Natural products 0.000 claims description 3
- CZMRCDWAGMRECN-UGDNZRGBSA-N Sucrose Chemical compound O[C@H]1[C@H](O)[C@@H](CO)O[C@@]1(CO)O[C@@H]1[C@H](O)[C@@H](O)[C@H](O)[C@@H](CO)O1 CZMRCDWAGMRECN-UGDNZRGBSA-N 0.000 claims description 3
- 235000019319 peptone Nutrition 0.000 claims description 3
- 239000005720 sucrose Substances 0.000 claims description 3
- 230000000694 effects Effects 0.000 abstract description 33
- 241000589155 Agrobacterium tumefaciens Species 0.000 abstract description 28
- 238000012360 testing method Methods 0.000 abstract description 25
- 239000003242 anti bacterial agent Substances 0.000 abstract description 15
- 244000005700 microbiome Species 0.000 abstract description 10
- 230000008485 antagonism Effects 0.000 abstract description 6
- 230000001965 increasing effect Effects 0.000 abstract description 6
- 238000006243 chemical reaction Methods 0.000 abstract description 3
- 238000009629 microbiological culture Methods 0.000 abstract description 3
- 238000002474 experimental method Methods 0.000 abstract description 2
- 238000005507 spraying Methods 0.000 abstract description 2
- 231100000588 tumorigenic Toxicity 0.000 abstract description 2
- 230000000381 tumorigenic effect Effects 0.000 abstract description 2
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 39
- 201000010099 disease Diseases 0.000 description 30
- 230000000844 anti-bacterial effect Effects 0.000 description 18
- 230000001580 bacterial effect Effects 0.000 description 18
- 235000013399 edible fruits Nutrition 0.000 description 14
- 230000003042 antagnostic effect Effects 0.000 description 13
- 229940088710 antibiotic agent Drugs 0.000 description 11
- 239000002689 soil Substances 0.000 description 10
- 239000000725 suspension Substances 0.000 description 10
- 238000004458 analytical method Methods 0.000 description 9
- 230000005764 inhibitory process Effects 0.000 description 9
- 241000186660 Lactobacillus Species 0.000 description 8
- 229940039696 lactobacillus Drugs 0.000 description 8
- 238000012216 screening Methods 0.000 description 8
- 239000000126 substance Substances 0.000 description 8
- 244000052616 bacterial pathogen Species 0.000 description 7
- 230000008901 benefit Effects 0.000 description 7
- 238000011161 development Methods 0.000 description 7
- 101150013736 gyrB gene Proteins 0.000 description 7
- 108020004465 16S ribosomal RNA Proteins 0.000 description 6
- DHVLDKHFGIVEIP-UHFFFAOYSA-N 2-bromo-2-(bromomethyl)pentanedinitrile Chemical compound BrCC(Br)(C#N)CCC#N DHVLDKHFGIVEIP-UHFFFAOYSA-N 0.000 description 6
- 239000005760 Difenoconazole Substances 0.000 description 6
- 230000001276 controlling effect Effects 0.000 description 6
- BQYJATMQXGBDHF-UHFFFAOYSA-N difenoconazole Chemical compound O1C(C)COC1(C=1C(=CC(OC=2C=CC(Cl)=CC=2)=CC=1)Cl)CN1N=CN=C1 BQYJATMQXGBDHF-UHFFFAOYSA-N 0.000 description 6
- 239000003899 bactericide agent Substances 0.000 description 5
- 238000004519 manufacturing process Methods 0.000 description 5
- 238000002360 preparation method Methods 0.000 description 5
- 238000011160 research Methods 0.000 description 5
- OICGFSUWXCJLCO-UHFFFAOYSA-N 1-ethylsulfonylsulfanylethane Chemical compound CCSS(=O)(=O)CC OICGFSUWXCJLCO-UHFFFAOYSA-N 0.000 description 4
- 244000077233 Vaccinium uliginosum Species 0.000 description 4
- 230000036541 health Effects 0.000 description 4
- 238000012163 sequencing technique Methods 0.000 description 4
- 244000063299 Bacillus subtilis Species 0.000 description 3
- 235000014469 Bacillus subtilis Nutrition 0.000 description 3
- 240000009088 Fragaria x ananassa Species 0.000 description 3
- 240000007594 Oryza sativa Species 0.000 description 3
- 235000007164 Oryza sativa Nutrition 0.000 description 3
- 230000007613 environmental effect Effects 0.000 description 3
- 244000053095 fungal pathogen Species 0.000 description 3
- 230000012010 growth Effects 0.000 description 3
- 230000006872 improvement Effects 0.000 description 3
- 101150012629 parE gene Proteins 0.000 description 3
- 230000001717 pathogenic effect Effects 0.000 description 3
- 239000000575 pesticide Substances 0.000 description 3
- 230000008635 plant growth Effects 0.000 description 3
- 238000000746 purification Methods 0.000 description 3
- 235000009566 rice Nutrition 0.000 description 3
- 239000000243 solution Substances 0.000 description 3
- 238000001228 spectrum Methods 0.000 description 3
- 241000589158 Agrobacterium Species 0.000 description 2
- 241000589156 Agrobacterium rhizogenes Species 0.000 description 2
- 208000035143 Bacterial infection Diseases 0.000 description 2
- 238000007400 DNA extraction Methods 0.000 description 2
- 238000010159 Duncan test Methods 0.000 description 2
- 235000016623 Fragaria vesca Nutrition 0.000 description 2
- 235000011363 Fragaria x ananassa Nutrition 0.000 description 2
- 206010018498 Goitre Diseases 0.000 description 2
- 240000008669 Hedera helix Species 0.000 description 2
- 241000831741 Lactobacillus farraginis Species 0.000 description 2
- 241000724205 Rice stripe tenuivirus Species 0.000 description 2
- 240000008042 Zea mays Species 0.000 description 2
- 235000005824 Zea mays ssp. parviglumis Nutrition 0.000 description 2
- 235000002017 Zea mays subsp mays Nutrition 0.000 description 2
- 235000005822 corn Nutrition 0.000 description 2
- 238000010790 dilution Methods 0.000 description 2
- 239000012895 dilution Substances 0.000 description 2
- 230000003631 expected effect Effects 0.000 description 2
- 238000000556 factor analysis Methods 0.000 description 2
- 235000013305 food Nutrition 0.000 description 2
- 238000007710 freezing Methods 0.000 description 2
- 230000008014 freezing Effects 0.000 description 2
- 230000006870 function Effects 0.000 description 2
- 201000003872 goiter Diseases 0.000 description 2
- 230000006799 invasive growth in response to glucose limitation Effects 0.000 description 2
- 239000000463 material Substances 0.000 description 2
- 244000000010 microbial pathogen Species 0.000 description 2
- 230000000877 morphologic effect Effects 0.000 description 2
- 238000013081 phylogenetic analysis Methods 0.000 description 2
- 230000008569 process Effects 0.000 description 2
- 239000000047 product Substances 0.000 description 2
- 239000008223 sterile water Substances 0.000 description 2
- 230000004083 survival effect Effects 0.000 description 2
- 238000010998 test method Methods 0.000 description 2
- 241000726119 Acidovorax Species 0.000 description 1
- 241001135511 Agrobacterium rubi Species 0.000 description 1
- 241000219193 Brassicaceae Species 0.000 description 1
- OKTJSMMVPCPJKN-UHFFFAOYSA-N Carbon Chemical compound [C] OKTJSMMVPCPJKN-UHFFFAOYSA-N 0.000 description 1
- 208000005623 Carcinogenesis Diseases 0.000 description 1
- 229920000858 Cyclodextrin Polymers 0.000 description 1
- 208000035240 Disease Resistance Diseases 0.000 description 1
- 239000004386 Erythritol Substances 0.000 description 1
- UNXHWFMMPAWVPI-UHFFFAOYSA-N Erythritol Natural products OCC(O)C(O)CO UNXHWFMMPAWVPI-UHFFFAOYSA-N 0.000 description 1
- 239000001116 FEMA 4028 Substances 0.000 description 1
- 241000233866 Fungi Species 0.000 description 1
- 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 1
- 238000003794 Gram staining Methods 0.000 description 1
- 229920002752 Konjac Polymers 0.000 description 1
- 235000007688 Lycopersicon esculentum Nutrition 0.000 description 1
- 241000863030 Lysobacter enzymogenes Species 0.000 description 1
- 241000244206 Nematoda Species 0.000 description 1
- 241000233654 Oomycetes Species 0.000 description 1
- 206010033157 Ovarian enlargement Diseases 0.000 description 1
- 241001659863 Panna Species 0.000 description 1
- 241000589516 Pseudomonas Species 0.000 description 1
- 241001646398 Pseudomonas chlororaphis Species 0.000 description 1
- 241001633102 Rhizobiaceae Species 0.000 description 1
- 240000004808 Saccharomyces cerevisiae Species 0.000 description 1
- 240000003768 Solanum lycopersicum Species 0.000 description 1
- 101100187186 Solanum lycopersicum LE16 gene Proteins 0.000 description 1
- 241000223259 Trichoderma Species 0.000 description 1
- 206010052428 Wound Diseases 0.000 description 1
- 208000027418 Wounds and injury Diseases 0.000 description 1
- 238000010521 absorption reaction Methods 0.000 description 1
- 230000003213 activating effect Effects 0.000 description 1
- 238000012271 agricultural production Methods 0.000 description 1
- 238000013459 approach Methods 0.000 description 1
- 238000003556 assay Methods 0.000 description 1
- 230000002358 autolytic effect Effects 0.000 description 1
- 208000022362 bacterial infectious disease Diseases 0.000 description 1
- WHGYBXFWUBPSRW-FOUAGVGXSA-N beta-cyclodextrin Chemical compound OC[C@H]([C@H]([C@@H]([C@H]1O)O)O[C@H]2O[C@@H]([C@@H](O[C@H]3O[C@H](CO)[C@H]([C@@H]([C@H]3O)O)O[C@H]3O[C@H](CO)[C@H]([C@@H]([C@H]3O)O)O[C@H]3O[C@H](CO)[C@H]([C@@H]([C@H]3O)O)O[C@H]3O[C@H](CO)[C@H]([C@@H]([C@H]3O)O)O3)[C@H](O)[C@H]2O)CO)O[C@@H]1O[C@H]1[C@H](O)[C@@H](O)[C@@H]3O[C@@H]1CO WHGYBXFWUBPSRW-FOUAGVGXSA-N 0.000 description 1
- 235000011175 beta-cyclodextrine Nutrition 0.000 description 1
- 229960004853 betadex Drugs 0.000 description 1
- 239000012681 biocontrol agent Substances 0.000 description 1
- 239000003124 biologic agent Substances 0.000 description 1
- 230000036952 cancer formation Effects 0.000 description 1
- 229910052799 carbon Inorganic materials 0.000 description 1
- 231100000504 carcinogenesis Toxicity 0.000 description 1
- 235000013339 cereals Nutrition 0.000 description 1
- 239000003795 chemical substances by application Substances 0.000 description 1
- MYSWGUAQZAJSOK-UHFFFAOYSA-N ciprofloxacin Chemical compound C12=CC(N3CCNCC3)=C(F)C=C2C(=O)C(C(=O)O)=CN1C1CC1 MYSWGUAQZAJSOK-UHFFFAOYSA-N 0.000 description 1
- 238000010227 cup method (microbiological evaluation) Methods 0.000 description 1
- 230000007547 defect Effects 0.000 description 1
- 238000010586 diagram Methods 0.000 description 1
- 238000003113 dilution method Methods 0.000 description 1
- 230000006806 disease prevention Effects 0.000 description 1
- 238000005516 engineering process Methods 0.000 description 1
- 230000002708 enhancing effect Effects 0.000 description 1
- 238000003912 environmental pollution Methods 0.000 description 1
- UNXHWFMMPAWVPI-ZXZARUISSA-N erythritol Chemical compound OC[C@H](O)[C@H](O)CO UNXHWFMMPAWVPI-ZXZARUISSA-N 0.000 description 1
- 235000019414 erythritol Nutrition 0.000 description 1
- 229940009714 erythritol Drugs 0.000 description 1
- 238000011049 filling Methods 0.000 description 1
- 239000008103 glucose Substances 0.000 description 1
- 150000004676 glycans Chemical class 0.000 description 1
- 238000000338 in vitro Methods 0.000 description 1
- 238000011534 incubation Methods 0.000 description 1
- 208000015181 infectious disease Diseases 0.000 description 1
- 238000011081 inoculation Methods 0.000 description 1
- 238000011835 investigation Methods 0.000 description 1
- 238000002955 isolation Methods 0.000 description 1
- 235000010485 konjac Nutrition 0.000 description 1
- 238000013332 literature search Methods 0.000 description 1
- 230000001320 lysogenic effect Effects 0.000 description 1
- 230000007246 mechanism Effects 0.000 description 1
- 239000002068 microbial inoculum Substances 0.000 description 1
- 230000002906 microbiologic effect Effects 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
- 235000015097 nutrients Nutrition 0.000 description 1
- 230000000050 nutritive effect Effects 0.000 description 1
- 244000052769 pathogen Species 0.000 description 1
- 230000003950 pathogenic mechanism Effects 0.000 description 1
- 230000007918 pathogenicity Effects 0.000 description 1
- 239000000447 pesticide residue Substances 0.000 description 1
- 230000008636 plant growth process Effects 0.000 description 1
- 238000007747 plating Methods 0.000 description 1
- 229920001282 polysaccharide Polymers 0.000 description 1
- 239000005017 polysaccharide Substances 0.000 description 1
- 230000008092 positive effect Effects 0.000 description 1
- 238000004382 potting Methods 0.000 description 1
- 230000001737 promoting effect Effects 0.000 description 1
- 108090000623 proteins and genes Proteins 0.000 description 1
- 230000009467 reduction Effects 0.000 description 1
- 230000001105 regulatory effect Effects 0.000 description 1
- 244000000000 soil microbiome Species 0.000 description 1
- 239000007921 spray Substances 0.000 description 1
- 235000021012 strawberries Nutrition 0.000 description 1
- 239000006228 supernatant Substances 0.000 description 1
- 208000024891 symptom Diseases 0.000 description 1
- 230000009466 transformation Effects 0.000 description 1
Landscapes
- Agricultural Chemicals And Associated Chemicals (AREA)
Abstract
The invention belongs to the technical field of microorganisms, and discloses an antibiotic lysobacter LMT16 and application thereof, wherein the antibiotic lysobacter LMT16 is preserved in the China general microbiological culture collection center (CGMCC) with the preservation number of 27317 in the 7 th month 18 th year 2023. Indoor plate antagonism experiments show that the antibiotic lysobacter LMT16 has obvious antagonism on the blueberry root cancer representative strain LM 11. The greenhouse control effect test shows that LMT16 can effectively inhibit the tumorigenic reaction of the agrobacterium tumefaciens LM11 of blueberry, and the greenhouse control effect can reach 77.50%. The strain antibiotic lysobacter LMT16 can be used for field prevention and treatment of blueberry root cancer by root irrigation and spraying, and has the effects of increasing yield and quality. The strain has bacteriostasis broad spectrum and wide application value.
Description
Technical Field
The invention belongs to the technical field of microorganisms, and particularly relates to an antibiotic lysobacter LMT16 and application thereof.
Background
Currently, the root cancer disease (crown gall), also known as crown gall disease, is a worldwide bacterial disease caused by Agrobacterium tumefaciens (Agrobacterium tumefaciens). Agrobacterium tumefaciens belongs to the genus Agrobacterium of the family Rhizobiaceae, and can be classified into Agrobacterium radiobacter (Agrobacterium radiobacter), agrobacterium tumefaciens (Agrobacterium tumefaciens), agrobacterium rhizogenes (Agrobacterium rhizogenes), and Agrobacterium rubrum (Agrobacterium rubi) depending on pathogenicity and host range. The agrobacterium tumefaciens has a wider host range, can infect 93 plants of 331 plants 643 of fruit trees, ornamental plants and the like, mainly damages plant roots, causes goiter to occur in the plant roots, seriously damages normal growth of the plants, causes reduced blueberry yield and reduced fruit quality, and can cause the death of the whole blueberry plant when the disease is serious.
Rhizosphere is one of the main sites for exchanging energy substances between plants and soil, and is also one of the main sites for microorganism propagation and survival, and in recent years, close relationship between rhizosphere microorganisms and plants has become an important point of attention and research of global researchers. Various types of microorganisms exist in rhizosphere soil, and certain microorganisms have negative effects in the plant growth process, so that energy conversion between plants and the rhizosphere soil can be influenced, slow plant growth is caused, and plant diseases can be caused; meanwhile, part of microorganisms can play a positive role in plant growth, can promote the absorption and transformation of plant root systems to nutrient components, can inhibit the growth and propagation of certain pathogenic microorganisms, such as trichoderma, lysobacter, pseudomonas, bacillus and the like, and can promote plant growth and inhibit the propagation of pathogenic microorganisms, which are called as bio-control microorganisms. Li Suping and other researches show that the autolytic lysogenic lysobacter enzymogenes LE16 separated from rhizosphere soil has a good inhibition effect on tomato gray mold; zhang Xitang and the like, a bacillus beijerinus JKXZ strain is separated from the root of sakura and used for preventing and treating sakura root cancer; the antibiotic lysobacter 136 isolated from the rhizosphere of konjak such as panna can effectively prevent and treat clubroot of cruciferae. Therefore, rhizosphere soil microorganisms have been paid attention to by many scholars in plant disease control, and therefore rhizosphere bacteria have a non-negligible important role as a selection object for a biocontrol agent.
Lysobacter is a type of gram-negative strain widely existing in nature, and can degrade various polysaccharides, and has a dissolving effect on pathogenic bacteria, yeasts, filamentous fungi, nematodes and oomycetes.
The lysobacter has the capability of preventing and treating various plant diseases, but the antibiotics of the lysobacter are not found yet through the document search, can inhibit the occurrence of blueberry root cancer caused by agrobacterium tumefaciens, and are applied to fields for preventing and treating blueberry root cancer.
Through the above analysis, the problems and defects existing in the prior art are as follows: root cancer disease occurs mainly in plant roots, and requires a long period of time from infection with pathogenic bacteria to the manifestation of symptoms in above-ground tissues. Once the plant is ill, it is difficult to control, and serious economic loss is caused. The root cancer is an important disease which damages the development of the blueberry industry in China, and the occurrence of different degrees of a plurality of blueberry planting bases in Yunnan causes the reduction of blueberry fruit quality or the death of plants, so that larger economic loss is caused, and the blueberry root cancer problem needs to be solved urgently. The current control of blueberry root cancer is mainly chemical control and agricultural control, the report of domestic biological control of blueberry root cancer is less, the pathogenic mechanism of the blueberry root cancer is not clear, the reported biological control strain test research is not applied in field scale, no new biological control preparation for controlling the blueberry root cancer is registered in China, the specific biological control mechanism of the biological control strain is not clear, the research is continued later, and safe and effective technical support and theoretical basis are provided for green control of blueberry root cancer.
Disclosure of Invention
Aiming at the problems existing in the prior art, the invention provides an antibiotic bacillus subtilis LMT16 for field prevention and treatment of blueberry root cancer.
The invention aims to provide an antibiotic lysobacter (Lysobacter antibioticus) LMT16, which is characterized in that the preservation number is CGMCC No.27317.
The invention further aims to provide an application of the lysobacter antibioticus LMT16 in preventing and treating blueberry root cancer.
Another object of the present invention is to provide a method for preventing and treating blueberry root cancer using the antibiotic bacillus subtilis LMT16, wherein the method for preventing and treating blueberry root cancer using the antibiotic bacillus subtilis LMT16 comprises the following steps:
firstly, transferring activated lysobacter antibioticus LMT16 to a solid flat plate culture medium, and culturing for 48 hours at constant temperature to obtain purified strains;
Step two, inoculating the obtained purified strain into a triangular flask of a liquid culture medium, and culturing in a constant-temperature shaking incubator to obtain seed liquid;
thirdly, transferring the seed liquid into a triangular flask filled with a liquid culture medium, and carrying out shaking culture to obtain a fermentation liquid for later use;
Fourthly, transplanting the 3-year-old blueberry plants into a pot, and carrying out root irrigation treatment on the plants by adopting the fermentation liquor prepared in the third step, wherein the dosage is 100 mL/plant, so that the prevention and treatment of blueberry root cancer are realized;
The formula of the solid culture medium in the first step is as follows: mannitol 1%, yeast extract 0.3%, beef extract 0.1%, dipotassium hydrogen phosphate 0.05%, sodium chloride 0.02%, magnesium sulfate 0.02%, agar powder 2%, distilled water to 1L, and pH to 7.0.
The formula of the liquid culture medium in the second step is as follows: mannitol 1%, yeast extract 0.3%, beef extract 0.1%, dipotassium hydrogen phosphate 0.05%, sodium chloride 0.02%, magnesium sulfate 0.02%, agar powder 2%, distilled water to 1L, and pH to 7.0.
In the second step, the obtained strain is inoculated into 100mL triangular flasks of the liquid culture medium, 50mL of the culture medium is filled in each flask, and the culture is carried out for 24 hours at the temperature of 28 ℃ in a constant-temperature shaking incubator at the speed of 150 r/min.
The formula of the liquid fermentation medium in the third step is as follows: 1% of industrial sucrose, 3% of industrial peptone, 0.0023% of dipotassium hydrogen phosphate and 0.02% of magnesium sulfate, distilled water is fixed to a volume of 1L, and the pH is adjusted to 6.0.
In the third step, the seed solution is transferred to a 1000mL triangular flask filled with 500mL of liquid culture medium, and the seed solution is cultured for 48h at 28 ℃ under 150r/min shaking.
Further, a fermentation broth of the lysobacter antibioticus LMT16 is obtained.
Further, the fermentation liquor is used as biocontrol bacteria to be applied to blueberry root cancer.
In combination with the technical scheme and the technical problems to be solved, the technical scheme to be protected has the following advantages and positive effects:
The first, the invention relates to a strain of antibiotic lysobacter and its application, belonging to the biotechnology field, its production strain is antibiotic lysobacter (Lysobacter antibioticus), named LMT16, and has been preserved in China general microbiological culture Collection center (CGMCC) No.27317 at 5-18 of 2023. The strain has broad antibacterial spectrum and strong antibacterial capability, has broad-spectrum antibacterial effect on pathogenic bacteria such as blueberry root cancer bacteria, rice stripe spot bacteria, hedera helix leaf spot bacteria and the like, and pathogenic fungi such as rice blast bacteria, corn small spot bacteria, strawberry gray mold bacteria and the like, and has inhibition rate of over 50 percent, wherein LMT16 has good inhibition effect on the tumorigenesis of blueberry root cancer bacteria, and has inhibition effect of over 82 percent, and the results show that the antibiotic lysobacter LMT16 has antibacterial broad-spectrum effect on various pathogenic bacteria and pathogenic fungi and has great application potential.
The invention can be used for preparing the biocontrol preparation for preventing and controlling diseases of various cash crops such as blueberries, strawberries, rice and the like, and has the characteristics of safety and high efficiency.
Second, the antibiotic bacillus provided by the invention can be used for preventing and treating blueberry root cancer, so that the damage of blueberry root cancer is reduced and the yield and quality of blueberry are improved.
The strain LMT16 has antagonism to various pathogenic bacteria, is easy to carry out industrialized production, and can be used for preparing biological agents for preventing and treating blueberry root cancer in fields.
The antibiotic lysobacter LMT16 biological preparation can be applied in root irrigation and spray mode to prevent and treat blueberry root cancer, has the functions of increasing yield and quality, and can be applied to field disease prevention and treatment.
The greenhouse test proves that the biological control bactericide can effectively inhibit plant goiter caused by agrobacterium tumefaciens, and the incidence degree is reduced. The disease index after LMT16 treatment was 8.33, which is much lower than that of control treatment 47.49. The diameter of the bacteriostasis circle of the strain LMT16 on the agrobacterium tumefaciens reaches 24mm, and the greenhouse control effect reaches 82.46%. The antagonistic bacteria were identified as lysobacter antibioticus by morphological features and alignment analysis of the 16S rRNA and gyrB gene sequences (Lysobacter antibioticus). These data are consistent with previous studies.
According to the invention, a field control effect test proves that the antibiotic lysobacter LMT16 has good field control effect on blueberry root cancer, and can effectively inhibit occurrence of new tumors of root systems of pathogenic plants. The incidence rate of new tumors of the blueberry plants treated by the LMT16 is 42.85 percent, which is lower than 56.25 percent of that of the control group; the disease plant value is 31.39 which is far lower than that of the control group 74.17, and the relative prevention effect can reach 57.65 percent. The blueberry plants treated by using LMT16 have fruit setting rate up to 80.76%, which is improved by 28.35% compared with the control group.
In order to excavate antagonistic bacterial strains and an effective method capable of performing biological control on blueberry root cancer, the invention adopts a flat plate antagonistic method to separate and screen effective antagonistic bacteria with in-vitro antibacterial activity on pathogenic root cancer bacteria from the rhizosphere soil of blueberry plants, and the classification status of the effective antagonistic bacteria is analyzed through morphological observation and 16S rRNA and gyrB gene sequencing; and then, carrying out statistics of prevention and control effects by using a potting test. Finally screening out a blueberry root cancer antagonistic bacterium LMT16 with the highest control effect, wherein the diameter of a bacteriostasis circle reaches 24mm; the antibiotics were identified by comparison analysis. The potted plant test result shows that: LMT16 significantly reduced the ability of agrobacterium tumefaciens to cause galls in blueberry plants compared to control. The field prevention and control effect of the lysobacter antibioticus LMT16 on the blueberry root cancer disease is up to 57.65 percent, and the yield and quality of blueberry fruits are obviously improved. The invention provides a new strategy for preventing and controlling blueberry root cancer establishment mode biological strains.
Thirdly, as inventive supplementary evidence of the claims of the present invention, the following important aspects are also presented:
The blueberry is used as a novel fruit with high economic value and high nutritive value, and has wide development prospect. Blueberry root cancer is currently one of the most important diseases in a Yunnan blueberry base, and is particularly difficult to control for soil-borne diseases. At present, chemical control and agricultural control are mainly used for blueberry root cancer, but a large amount of chemical pesticides are easy to cause pesticide residue, environmental pollution and the like, so that the quality and yield of fruits are reduced, and the biological control of blueberry root cancer becomes a main research hot spot. There is no report of field prevention and treatment of blueberry root cancer by using antibiotics, and the invention makes clear a kind of biocontrol bacteria with obvious inhibition effect on blueberry root cancer, evaluates the greenhouse and field prevention effect, explores the prevention effect of the antibiotics on blueberry root cancer, and provides safe and effective technical support and theoretical basis for green prevention and control of blueberry root cancer.
In our province, root cancer disease commonly occurs on blueberry plants and is seriously damaged. Along with the expansion of the cultivation area of blueberry fruit trees and the increase of the number of planting years, serious economic loss occurs in the blueberry industry. At present, the method for preventing and treating the blueberry root cancer mainly comprises a combination of chemical prevention and treatment and agricultural prevention and treatment. Through literature search, public reports of preventing and treating blueberry dry cancer diseases by using the antibiotics lysobacter are not yet seen and applied to fields. The lysobacter antibioticus LMT16 has wide bacteriostasis spectrum and can prevent and treat various bacterial diseases. The technical scheme of the invention is expected to solve the damage of blueberry root cancer, reduce economic loss, have great commercial value and have good development prospect.
Fourth, the technology for preventing and treating blueberry root cancer disease by using the lysobacter antibioticus LMT16 (preservation number CGMCCNo.27317) provided by the invention brings about several remarkable technical progress:
1) The innovative biological control method comprises the following steps:
The use of specific microorganisms such as LMT16 for disease control represents an environmentally friendly and sustainable biological control method compared to traditional chemical pesticides. This approach reduces the use of chemicals and reduces the negative environmental impact.
2) Safety of blueberry production is improved:
The use of the lysobacter antibioticus LMT16 as a biological control agent can effectively reduce chemical residues, thereby improving the food safety of blueberry products. This is of great importance for maintaining consumer health and improving market trust.
3) Enhancing disease resistance of blueberry plants:
The application of LMT16 can prevent and treat root cancer, strengthen the whole health and growth vigor of plants and improve the resistance and productivity of the plants.
4) Improvement of economic benefit:
As LMT16 can effectively control the root cancer disease, it is helpful for reducing the economic loss caused by the disease and increasing the economic benefit of blueberry planting.
5) The operation is simple and convenient, and the popularization is easy:
the culture and application processes of the LMT16 are simple and convenient, and the LMT16 is easy to be produced and popularized and used on a large scale. For blueberry growers, the method is simple to operate and easy to accept.
6) Promote sustainable development of agriculture:
The adoption of the biological control method accords with the sustainable development trend of agriculture, and is helpful for promoting the development of modern agriculture towards more environmental protection and health.
In general, the method for preventing and treating blueberry root cancer by using the lysobacter antibioticus LMT16 not only provides an effective biological prevention and treatment means, but also brings remarkable technical progress in the aspects of environmental protection, food safety, economic benefit, agricultural sustainable development and the like.
Drawings
FIG. 1 is a flowchart of an application method of the lysobacter antibioticus LMT16 provided by the embodiment of the invention;
FIG. 2 is a schematic diagram of phylogenetic analysis based on antagonistic bacterial strain 16S rRNA (A of FIG. 2) and gyrB (B of FIG. 2) sequences provided by the examples of the present invention;
FIG. 3 is a schematic representation of colonies provided by an embodiment of the present invention.
Detailed Description
The present invention will be described in further detail with reference to the following examples in order to make the objects, technical solutions and advantages of the present invention more apparent. It should be understood that the specific embodiments described herein are for purposes of illustration only and are not intended to limit the scope of the invention.
Example 1: control application of small-scale blueberry garden
1) Culturing strains: in a laboratory environment, the antibiotic lysobacter LMT16 was transferred onto a solid plate medium and cultured at a constant temperature of 28 ℃ for 48 hours to obtain a strain.
2) Seed liquid and fermentation liquid preparation:
inoculating the strain into a triangular flask of a liquid culture medium, and culturing in a constant-temperature shaking incubator to obtain seed liquid. The seed liquid is further transferred into a triangular flask filled with a liquid culture medium, and the seed liquid is cultured by shaking, so that a fermentation liquid for field application is obtained.
3) And (3) field application: selecting 3-year-old blueberry plants, and transplanting the blueberry plants into a basin. And (3) root irrigation treatment is carried out on blueberry plants by using fermentation liquor, wherein the dosage of each plant is about 100mL, so as to prevent and treat the root cancer.
The expected effect is: effectively control the occurrence of small-scale blueberry garden root cancer and improve the yield and quality of blueberries.
Example 2: comprehensive management of large blueberry planting base
1) Culturing the strain on a large scale:
in the industrialized fermentation facility, LMT16 strains are cultivated in an enlarged mode according to the method so as to meet the requirement of large-area planting.
2) Mass production of fermentation liquor:
large fermenters are used to produce fermentation broth, ensuring a sufficient supply.
3) The field irrigation system is applied: the fermentation broth was distributed to each blueberry plant by an existing field irrigation system. And (3) regularly irrigating the blueberry garden to prevent and treat the root cancer.
The expected effect is: under the condition of large-area planting, the spread of the root cancer is effectively controlled, the overall planting benefit is improved, and meanwhile, the health of soil and ecological environment is maintained.
These two examples demonstrate the use of the antibiotic lysobacter antibioticus LMT16 in different scale blueberry planting, from small horticultural applications to large agricultural production, both showing its potential and utility in biological control of root cancer.
The lysobacter antibioticus (Lysobacter antibioticus) LMT16 provided by the embodiment of the invention is separated from blueberry rhizosphere soil of modern agriculture limited company of Qu Jingshi Jiawa in Yunnan province;
The strain has the following characteristics:
(1) Colonies appeared yellow brown, circularly transparent, with flat and glossy middle raised edges (fig. 3). As the incubation time increases, the color of the colonies also deepens.
(2) Has the following physiological and biochemical characteristics: gram staining is negative, and 32 carbon sources such as i erythritol, beta cyclodextrin, aD glucose, D mannitol and the like can be decomposed and utilized.
(3) The strain has broad antibacterial spectrum and strong antibacterial capability, has broad-spectrum antibacterial effect on pathogenic bacteria such as blueberry root cancer bacteria, rice stripe spot bacteria, hedera helix leaf spot bacteria and the like, and pathogenic fungi such as rice blast bacteria, corn small spot bacteria, strawberry gray mold bacteria and the like, and has inhibition rate of more than 50%.
As shown in FIG. 1, the application method of the lysobacter antibioticus LMT16 provided by the embodiment of the invention comprises the following steps:
s101: transferring the lysobacter antibioticus LMT16 to a solid flat-plate culture medium, and culturing for 48 hours at the constant temperature of 28 ℃ to obtain strains;
s102: inoculating the obtained strain into 100mL triangular flasks of the liquid culture medium, filling 50mL of the triangular flasks each time, and culturing for 24 hours at the temperature of 28 ℃ in a constant-temperature shaking incubator at the speed of 150r/min to obtain seed liquid;
S103: transferring the seed liquid into a 1000mL triangular flask filled with 500mL of liquid culture medium, and culturing at 28 ℃ under shaking for 48h at 150r/min to obtain fermentation liquor for later use;
S104: transplanting 3-year-old blueberry plants into a pot, and performing root irrigation treatment on the plants by adopting the fermentation liquor prepared in the step S3, wherein the dosage is 100 mL/plant, so that the prevention and treatment of blueberry root cancer can be realized.
In an embodiment of the present invention, the solid medium formulation in S101 is: mannitol 1%, yeast extract 0.3%, beef extract 0.1%, dipotassium hydrogen phosphate 0.05%, sodium chloride 0.02%, magnesium sulfate 0.02%, agar powder 2%, distilled water to 1L, and pH to 7.0.
In an embodiment of the present invention, the liquid medium formulation in S102 is: mannitol 1%, yeast extract 0.3%, beef extract 0.1%, dipotassium hydrogen phosphate 0.05%, sodium chloride 0.02%, magnesium sulfate 0.02%, agar powder 2%, distilled water to 1L, and pH to 7.0.
In an embodiment of the present invention, the liquid fermentation medium formulation in S103 is: 1% of industrial sucrose, 3% of industrial peptone, 0.0023% of dipotassium hydrogen phosphate and 0.02% of magnesium sulfate, distilled water is fixed to a volume of 1L, and the pH is adjusted to 6.0.
In an embodiment of the invention, a fermentation broth of the lysobacter antibioticus LMT16 is obtained.
In the embodiment of the invention, the fermentation liquor is used as biocontrol bacteria for treating blueberry root cancer.
The lysobacter antibioticus LMT16 is preserved in China general microbiological culture Collection center (CGMCC) at the 18 th month of 2023, and has the address of North Star Xiyu No. 1, 3 rd, china academy of sciences microbiological study, and the preservation number of CGMCC No.27317. Indoor plate antagonism experiments show that the antibiotic lysobacter LMT16 has obvious antagonism on the blueberry root cancer representative strain LM 11. The greenhouse control effect test shows that LMT16 can effectively inhibit the tumorigenic reaction of the agrobacterium tumefaciens LM11 of blueberry, and the greenhouse control effect can reach 77.50%. The strain antibiotic lysobacter LMT16 can be used for field prevention and treatment of blueberry root cancer by root irrigation and spraying, and has the effects of increasing yield and quality. The strain has bacteriostasis broad spectrum and wide application value.
Example 1 isolation and purification of blueberry rhizosphere soil bacteria
Blueberry rhizosphere soil is collected from different blueberry bases of Qu Jingjia Wo modern agriculture limited company, and antagonistic bacteria are separated by adopting a gradient dilution method. 1g of rhizosphere soil is weighed, transferred into a triangular flask, 9mL of sterile water is added, shaking culture is carried out for 30min at the temperature of 30 ℃ and the speed of 150r/min, the supernatant is subjected to gradient dilution (10 -3～10-6), and 100 mu L of each is coated and separated on YEM and NA culture medium according to low concentration to high concentration. And (3) placing the coated culture dish in a 28 ℃ incubator for 2d, picking different single colonies on the culture medium for purification culture, picking single bacterial colonies after two generations of purification, inoculating the single bacterial colonies into an NA freezing tube, and placing the NA freezing tube in a refrigerator at the temperature of minus 20 ℃ for later use.
Example 2 screening of antagonistic strains of Agrobacterium tumefaciens
1 Test material
1.1 Test strains
Lysobacter antibioticus (Lysobacter antibioticus) LMT16; agrobacterium tumefaciens (Agrobacterium tumefaciens) LM11 (supplied by the university of agricultural bacteria, yunnan).
1.2 Test Medium
TABLE 1 test Medium formulation
2 Test method
2.1 Preliminary screening of antagonistic strains
Screening the separated bacteria by using the agrobacterium tumefaciens LM11 as a target bacteria by a spot grafting method, inoculating the purified LM11 into YEB (YEM liquid culture medium), and culturing at 30 ℃ for 24 hours under shaking at 150r/min to prepare pathogenic bacteria suspension. 1mL of the bacterial suspension is diluted to OD 600nm =0.30, 100 mu L of the bacterial suspension is added into a YEM plate, and the bacterial suspension is completely and uniformly smeared by a sterilized coater to prepare the bacteria-carrying YEM plate. Activating the bacteria in the NA plate, drawing lines, picking single bacterial drop points, inoculating the single bacterial drop points on the bacteria-carrying plate, inoculating 3 bacterial strains on each plate point, and repeating for 3 times. Culturing at 28deg.C for 48 hr, observing colony morphology and size of inhibition zone, and measuring diameter (mm) of inhibition zone.
2.2 Rescreening of antagonistic strains
And (3) selecting biocontrol strains with obvious inhibition zones in the primary screening result for re-screening, preparing a bacteria-containing plate of LM11 by referring to the method, transferring 200 mu L of biocontrol bacteria suspension prepared in advance into an oxford cup by adopting an oxford cup method, and repeating each treatment for 3 times. Culturing at 28deg.C for 48 hr, observing colony morphology and size of antibacterial zone, measuring diameter (mm) of antibacterial zone, and selecting strain with maximum antibacterial zone as the test biocontrol bacterium for subsequent test.
Separating 77 bacterial strains from different samples by a plate dilution plating method, performing a plate antagonism test with the agrobacterium tumefaciens LM11, and screening 6 bacterial strains with antibacterial activity on the agrobacterium tumefaciens. Further, the re-screening shows that the LMT16 has the strongest antibacterial activity on the agrobacterium tumefaciens, the diameter of a bacteriostasis zone reaches 24mm, and the diameters of other bacterial strains are 1217 mm.
EXAMPLE 3 identification of antagonistic strains of Agrobacterium tumefaciens
Total DNA extraction was performed according to the instructions using the antagonistic strain LMT16 as a template using a bacterial genomic DNA extraction kit. The multi-gene joint identification is carried out on the biocontrol bacteria by using 16S rRNA and gyrB genes, wherein Y=C/T, R=A/G and N=A/G/C/T in degenerate primers of the gyrB genes, and LMT16 genomic DNA is amplified by a PCR instrument. The product was sent to biosystems for sequencing. BLAST comparison analysis is carried out on the sequencing result, the 16S rRNA and gyrB gene sequences of related strains are obtained, phylogenetic analysis is carried out by MEGAX, and a phylogenetic tree is constructed by adopting an adjacent method, as shown in figure 2. BLAST comparison analysis is carried out on the sequencing result, and the 16S rRNA gene sequence homology analysis result shows that the LMT16 has the highest homology with the lysobacter antibiotic. The gyrB gene sequence homology analysis result shows that LMT16 has the highest homology with the antibiotics lysobacter. LMT16 was therefore determined to be the antibiotic lysobacter (Lysobacter antibioticus).
Example 4 greenhouse disease control Effect of Acidovorax faciens LMT16 on blueberry root canceration bacteria
Test strain: lysobacter antibioticus (Lysobacter antibioticus) LMT16; lactobacillus situm (Lactobacillus farraginis) Lf-8; agrobacterium tumefaciens (Agrobacterium tumefaciens) LM11.
The antibiotics lysobacter LMT16, the chaff lactobacillus Lf-8 and the pathogenic bacteria agrobacterium tumefaciens LM11 are respectively inoculated into YH, MRS and YEM liquid culture media, are subjected to shaking culture for 48 hours at 28 ℃ and 150rpm, and are regulated into bacterial suspension with the concentration of about 10 8 CFU/mL by using sterile water for standby.
The present embodiment sets 4 processes: (1) biocontrol bacteria treatment: transplanting blueberry seedlings growing for 2 years into a flowerpot, and uniformly pouring the suspension of the antibiotics lysobacter LMT16 and the chaff lactobacillus Lf8 into the root of the potted blueberry seedlings after 7d, wherein each pot is irrigated with 200mL. Inoculating the blueberry agrobacterium tumefaciens LM11 bacterial suspension to the roots of blueberry seedlings by a root irrigation method after 24 hours, and irrigating 200mL of the blueberry seedlings in each pot. (2) pesticide treatment: uniformly pouring 25% of equal volume of difenoconazole bromothalonil (1000 times of liquid) into the root of a potted blueberry seedling, and inoculating an equal volume of a suspension of the agrobacterium tumefaciens LM11 into the potted blueberry seedling by a root irrigation method after 24 hours. (3) control group: and (3) pouring an equal volume of water into the roots of the blueberry seedlings, and inoculating the blueberry agrobacterium tumefaciens LM11 bacterial suspension into the blueberry seedlings by a root pouring method after 24 hours. The root and stem bases of blueberries were scratched and wounds were made on the roots at the time of pathogen inoculation, 10 seedlings were treated each, and the incidence and disease index of the blueberry seedlings were investigated after 4 months.
Grading according to the grading standard of blueberry root cancer (see table 2), and calculating the morbidity, disease index and prevention effect of each treatment according to the following formula.
Incidence = (number of survey plants/total number of survey plants) ×100%
Disease index = [ Σ (number of disease stages×number of stages represented)/(total number of strains×number of highest stages represented value) ]100
Control = (1-disease index of control group/disease index of test group) ×100%
Table 2 blueberry root cancer disease grading criteria
Test results and analysis: as shown in Table 3, compared with 25% of difenoconazole bromothalonil (1000 times liquid) and clear water, the incidence rate and the disease index of the blueberry seedling root cancer treated by the antibiotics lysobacter LMT16 are obviously reduced, and compared with the other biocontrol strain of the cereal bran lactobacillus Lf8, the control efficiency is higher, which indicates that the biocontrol effect of the antibiotics lysobacter LMT11 is excellent and stable, and the greenhouse control effect on the blueberry root cancer is good.
Table 3 greenhouse control of LMT16 on blueberry root cancer
Note that: the different letters in the table from the same column indicate a significant level of difference of 0.05.
Example 5 field control of lysobacter antibiotic LMT16 against blueberry root cancer
Test material: lysobacter antibioticus (Lysobacter antibioticus) LMT16; lactobacillus situm (Lactobacillus farraginis) Lf-8;25% of difenoconazole bromothalonil WP;80% ethylicin EC
Blueberry variety: "Jewelry".
Test site: qu Jingshi Wo modern agriculture Co., ltd.
The test method comprises the following steps: the antibiotics of lysobacter LMT16 and Lactobacillus glutinosa Lf8 are used as the test biocontrol bacteria, 25% of difenoconazole bromothalonil WP and 80% of ethylicin EC are used as the test bactericides (see table 4), and clear water is used as a control. Blueberry plants of the base E3 cell were selected for the efficacy control assay, 20 blueberry plants per treatment, 3 replicates per treatment. The test strain is fermented and cultured in advance to obtain fermentation liquor, the fermentation liquor is diluted by 20 times, blueberry plants are irrigated with roots, and the test bactericide is diluted according to recommended concentration and irrigated with roots. Root irrigation is started on 10 months and 15 days, 2L of each blueberry seedling is irrigated, and root irrigation is carried out continuously for 5 times at intervals of 30d. The fruit setting rate of blueberries is investigated in the beginning of 4 months, the hundred-grain weights of 3 blueberries are investigated in 5 months, and the incidence rate and the disease index of new tumors at the root of blueberry root cancer plants are investigated and treated and controlled in the end of 7 months.
Grading according to the grading standard of blueberry root cancer (see table 3), and calculating the fruit setting rate, morbidity, disease index and prevention effect of each treatment according to the following formula.
Fruit setting rate = (total number of ovary enlarged fruits/total number of single flowers) ×100%
New tumor incidence = number of new tumor lines/total number of investigation x 100%
Disease index = [ Σ (number of disease stages×number of stages represented)/(total number of strains×number of highest stages represented value) ]100
Relative control effect = [ (control group disease index treatment group disease index)/control group disease index ] ×100%
TABLE 4 different microbial inoculum and use concentration of the bactericide
Test results and analysis: the test results are shown in tables 5 and 6, and the table 5 and the table 6 show that 4 different treatments have different degrees of control effects on blueberry root cancer, and the disease index is reduced by more than 40 relative to the control after LMT16 root irrigation treatment, and the control effect is up to 57.65%. Compared with biocontrol bacteria Lf8 and two bactericides, the LMT16 has more advantages in reducing occurrence of new tumors of blueberry root cancer plants and controlling disease conditions. And the blueberry plant root irrigated by using LMT16 has certain improvement effect on fruit yield and quality, and the yield of the blueberry plant can be improved by 680.67 g/plant. The biological control bacterial agent, namely the lysobacter antibioticus LMT16, has stable and better control effect and yield increasing effect on blueberry root cancer diseases, and simultaneously has the basic condition of mass popularization in production.
Table 5 field control Effect of LMT16 on blueberry root cancer
Note that: adopting Duncan test in single factor analysis of variance, p is less than 0.05; t1: the antibiotic lysobacter LMT16; t2: lactobacillus situm Lf8; t3:25% of difenoconazole bromothalonil; t4:80% ethylicin; CK: and (3) controlling.
TABLE 6 influence of LMT16 on blueberry fruit quality and yield
Note that: adopting Duncan test in single factor analysis of variance, p is less than 0.05; t1: the antibiotic lysobacter LMT16; t2: lactobacillus situm Lf8; t3:25% of difenoconazole bromothalonil; t4:80% ethylicin; CK: and (3) controlling.
The overall result shows that the biological preparation of the lysobacter antibioticus LMT16 can effectively prevent and treat blueberry root cancer, has the functions of increasing yield and quality, and can be applied to field survival.
The foregoing is merely illustrative of specific embodiments of the present invention, and the scope of the invention is not limited thereto, but any modifications, equivalents, improvements and alternatives falling within the spirit and principles of the present invention will be apparent to those skilled in the art within the scope of the present invention.
Claims (10)
1. The lysobacter antibioticus LMT16 is characterized in that the lysobacter antibioticus LMT16 has a preservation number of CGMCC No.27317.
2. Use of the antibiotic lysobacter LMT16 according to claim 1 for the prevention and treatment of blueberry root cancer.
3. A method of using the lysobacter antibioticus LMT16 as claimed in claim 2 for the control of blueberry root cancer, wherein the method of using the lysobacter antibioticus LMT16 for the control of blueberry root cancer comprises the steps of:
Firstly, transferring the lysobacter antibioticus LMT16 to a solid flat-plate culture medium, and culturing for 48 hours at the constant temperature of 28 ℃ to obtain strains;
Step two, inoculating the obtained strain into a triangular flask of a liquid culture medium, and culturing in a constant-temperature shaking incubator to obtain seed liquid;
thirdly, transferring the seed liquid into a triangular flask filled with a liquid culture medium, and carrying out shaking culture to obtain a fermentation liquid for later use;
And fourthly, transplanting the 3-year-old blueberry plants into a pot, and carrying out root irrigation treatment on the plants by adopting the fermentation liquor prepared in the third step, wherein the dosage is 100 mL/plant, so that the prevention and treatment of blueberry root cancer are realized.
4. The method for preventing and treating blueberry root cancer using the antibiotic lysobacter LMT16 as claimed in claim 3, wherein the solid medium formulation in the first step is: mannitol 1%, yeast extract 0.3%, beef extract 0.1%, dipotassium hydrogen phosphate 0.05%, sodium chloride 0.02%, magnesium sulfate 0.02%, agar powder 2%, distilled water to 1L, and pH to 7.0.
5. The method for preventing and treating blueberry root cancer using the antibiotic lysobacter LMT16 as claimed in claim 3, wherein the liquid medium formulation in the second step is: mannitol 1%, yeast extract 0.3%, beef extract 0.1%, dipotassium hydrogen phosphate 0.05%, sodium chloride 0.02%, magnesium sulfate 0.02%, agar powder 2%, distilled water to 1L, and pH to 7.0.
6. The method for preventing and treating blueberry root cancer using the antibiotic-containing bacillus LMT16 as set forth in claim 3, wherein in the second step, the obtained strain is inoculated into 100mL triangular flasks of the liquid culture medium, 50mL of the culture medium is packed in each flask, and the culture is carried out for 24 hours at the temperature of 28 ℃ in a constant temperature shaking incubator at 150 r/min.
7. The method for preventing and treating blueberry root cancer using the antibiotic lysobacter LMT16 as claimed in claim 3, wherein the liquid fermentation medium formula in the third step is: 1% of industrial sucrose, 3% of industrial peptone, 0.0023% of dipotassium hydrogen phosphate and 0.02% of magnesium sulfate, distilled water is fixed to a volume of 1L, and the pH is adjusted to 6.0.
8. The method for preventing and treating blueberry root cancer using the antibiotic-containing bacillus LMT16 as set forth in claim 3, wherein the third step is to transfer the seed solution into a 1000mL triangular flask containing 500mL of liquid culture medium, and shake-culture at 28 ℃ for 48h at 150 r/min.
9. The method for the use of the lysobacter antibioticus LMT16 for the prevention and treatment of blueberry root cancer according to claim 3, wherein a fermentation broth of the lysobacter antibioticus LMT16 is obtained.
10. The method for the use of the antibiotic lysobacter LMT16 for the prevention and treatment of blueberry root cancer according to claim 9, wherein the fermentation broth is used as a biocontrol bacterium for the use in blueberry root cancer.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202410106440.9A CN118086105A (en) | 2024-01-25 | 2024-01-25 | Lysobacter antibioticus LMT16 and application thereof |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202410106440.9A CN118086105A (en) | 2024-01-25 | 2024-01-25 | Lysobacter antibioticus LMT16 and application thereof |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN118086105A true CN118086105A (en) | 2024-05-28 |
Family
ID=91143370
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202410106440.9A Pending CN118086105A (en) | 2024-01-25 | 2024-01-25 | Lysobacter antibioticus LMT16 and application thereof |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN118086105A (en) |
-
2024
- 2024-01-25 CN CN202410106440.9A patent/CN118086105A/en active Pending
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN110616171B (en) | Saline-alkali-resistant Pacific bacillus and viable bacteria preparation and application thereof | |
| CN111349590B (en) | Bacillus amyloliquefaciens and application thereof in prevention and treatment of potato late blight | |
| CN104560827B (en) | A kind of biocontrol actinomycetes bacterial strain for preventing and treating tobacco bacterial wilt and its application | |
| CN111172063B (en) | Bacillus beleisi MES861 and application and product thereof | |
| CN107136122B (en) | Biocontrol microbial inoculum for preventing and treating potato late blight | |
| WO2021073142A1 (en) | Strain of fluorescent pseudomonas 22g5 and application thereof in preventative treatment of verticillium wilt in crops | |
| CN109112069B (en) | Biocontrol endophytic fungus and application thereof | |
| CN111073827B (en) | Bacillus beilesensis JTB8-2 and application thereof in biological control of orobanche meloidogyne | |
| KR20130056585A (en) | Plant growth promotion by using bacterial strains isolated from roots of miscanthus sacchariflorus | |
| CN108220210A (en) | One plant of Antagonistic Fungi Z-18 for preventing cotton verticillium wilt and application | |
| CN108587969B (en) | Preparation and application of verticillium dahliae strain HCX-01 capable of preventing and treating cotton verticillium wilt | |
| CN107325980A (en) | A kind of radiation hardness series bacillus KH9 and its application in biological antitranspirant | |
| CN117467557A (en) | Paenibacillus strain with growth promoting and biocontrol functions and application thereof | |
| CN112322559B (en) | Peanut endophytic bacterium bacillus subtilis F-1 and application thereof | |
| CN112825864B (en) | Application of Paenibacillus jimila in preventing and treating tomato bacterial wilt | |
| CN118086105A (en) | Lysobacter antibioticus LMT16 and application thereof | |
| CN117431191B (en) | Bacillus pseudomycoides HDCP1 and application thereof | |
| CN115232777B (en) | Streptomyces pseudolightyellow bacterial agent for preventing and treating tobacco black shank and application thereof | |
| CN116426445B (en) | Pseudomonas bacteria NJAU-T102 and application thereof | |
| CN113278551B (en) | Burkholderia, bacterial agent comprising Burkholderia, bacterial fertilizer, preparation method and application | |
| CN114164126B (en) | Rhizopus oryzae XERF-1 and application thereof in relieving apple continuous cropping obstacle | |
| CN115851534B (en) | Bacillus belicus strain for tobacco rhizosphere and application of bacillus belicus strain in prevention and control of tobacco diseases | |
| CN117800794B (en) | Microbial fertilizer for promoting growth of purple perilla and accumulation of medicinal components and application thereof | |
| CN115927051B (en) | Biocontrol bacterium, biocontrol compound microbial inoculum and application thereof | |
| CN108085284B (en) | Bacillus methylotrophicus, and preparation method and application of microbial inoculum containing same |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination |