CN114958660A - Paenibacillus polymyxa strain and application thereof - Google Patents
Paenibacillus polymyxa strain and application thereof Download PDFInfo
- Publication number
- CN114958660A CN114958660A CN202210520768.6A CN202210520768A CN114958660A CN 114958660 A CN114958660 A CN 114958660A CN 202210520768 A CN202210520768 A CN 202210520768A CN 114958660 A CN114958660 A CN 114958660A
- Authority
- CN
- China
- Prior art keywords
- paenibacillus polymyxa
- strain
- culture
- preparation
- aqueous solution
- 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
- 241000194105 Paenibacillus polymyxa Species 0.000 title claims abstract description 92
- 238000000855 fermentation Methods 0.000 claims abstract description 37
- 230000004151 fermentation Effects 0.000 claims abstract description 37
- 238000012360 testing method Methods 0.000 claims abstract description 34
- 238000002360 preparation method Methods 0.000 claims abstract description 26
- 235000003143 Panax notoginseng Nutrition 0.000 claims abstract description 25
- 241000180649 Panax notoginseng Species 0.000 claims abstract description 25
- 235000013311 vegetables Nutrition 0.000 claims abstract description 24
- 239000000725 suspension Substances 0.000 claims abstract description 23
- 239000007864 aqueous solution Substances 0.000 claims abstract description 22
- 238000011081 inoculation Methods 0.000 claims abstract description 20
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims abstract description 18
- 239000007788 liquid Substances 0.000 claims abstract description 17
- 244000005700 microbiome Species 0.000 claims abstract description 10
- 201000010099 disease Diseases 0.000 claims description 52
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 claims description 52
- 239000001963 growth medium Substances 0.000 claims description 32
- 238000000034 method Methods 0.000 claims description 22
- 238000012258 culturing Methods 0.000 claims description 18
- 239000002609 medium Substances 0.000 claims description 13
- PEDCQBHIVMGVHV-UHFFFAOYSA-N Glycerine Chemical compound OCC(O)CO PEDCQBHIVMGVHV-UHFFFAOYSA-N 0.000 claims description 12
- 244000061458 Solanum melongena Species 0.000 claims description 12
- 235000002597 Solanum melongena Nutrition 0.000 claims description 12
- CSNNHWWHGAXBCP-UHFFFAOYSA-L Magnesium sulfate Chemical compound [Mg+2].[O-][S+2]([O-])([O-])[O-] CSNNHWWHGAXBCP-UHFFFAOYSA-L 0.000 claims description 8
- 239000001888 Peptone Substances 0.000 claims description 8
- 108010080698 Peptones Proteins 0.000 claims description 8
- 235000019319 peptone Nutrition 0.000 claims description 8
- 230000001954 sterilising effect Effects 0.000 claims description 8
- WQZGKKKJIJFFOK-GASJEMHNSA-N Glucose Natural products OC[C@H]1OC(O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-GASJEMHNSA-N 0.000 claims description 7
- 229940041514 candida albicans extract Drugs 0.000 claims description 7
- 239000012153 distilled water Substances 0.000 claims description 7
- 239000008103 glucose Substances 0.000 claims description 7
- 238000003973 irrigation Methods 0.000 claims description 7
- 230000002262 irrigation Effects 0.000 claims description 7
- 239000012138 yeast extract Substances 0.000 claims description 7
- 230000008569 process Effects 0.000 claims description 6
- 244000131316 Panax pseudoginseng Species 0.000 claims description 5
- 235000003181 Panax pseudoginseng Nutrition 0.000 claims description 5
- 229920001817 Agar Polymers 0.000 claims description 4
- 239000008272 agar Substances 0.000 claims description 4
- 235000015278 beef Nutrition 0.000 claims description 4
- ZPWVASYFFYYZEW-UHFFFAOYSA-L dipotassium hydrogen phosphate Chemical compound [K+].[K+].OP([O-])([O-])=O ZPWVASYFFYYZEW-UHFFFAOYSA-L 0.000 claims description 4
- 239000000284 extract Substances 0.000 claims description 4
- 229910052943 magnesium sulfate Inorganic materials 0.000 claims description 4
- 235000019341 magnesium sulphate Nutrition 0.000 claims description 4
- 239000000843 powder Substances 0.000 claims description 4
- 238000004659 sterilization and disinfection Methods 0.000 claims description 4
- 238000003756 stirring Methods 0.000 claims description 4
- 238000003672 processing method Methods 0.000 claims description 3
- 238000011218 seed culture Methods 0.000 claims description 3
- 239000000758 substrate Substances 0.000 claims description 3
- 241001638069 Rigidoporus microporus Species 0.000 claims 1
- 235000010149 Brassica rapa subsp chinensis Nutrition 0.000 abstract description 21
- 235000000536 Brassica rapa subsp pekinensis Nutrition 0.000 abstract description 21
- 241000499436 Brassica rapa subsp. pekinensis Species 0.000 abstract description 21
- 240000007124 Brassica oleracea Species 0.000 abstract description 16
- 235000003899 Brassica oleracea var acephala Nutrition 0.000 abstract description 15
- 235000011301 Brassica oleracea var capitata Nutrition 0.000 abstract description 15
- 235000001169 Brassica oleracea var oleracea Nutrition 0.000 abstract description 15
- 238000004321 preservation Methods 0.000 abstract description 4
- -1 inoculation Substances 0.000 abstract description 2
- 230000000694 effects Effects 0.000 description 29
- 230000000443 biocontrol Effects 0.000 description 28
- 230000001580 bacterial effect Effects 0.000 description 22
- 241000894006 Bacteria Species 0.000 description 19
- 241000196324 Embryophyta Species 0.000 description 19
- 244000052616 bacterial pathogen Species 0.000 description 19
- 238000011282 treatment Methods 0.000 description 17
- 239000002068 microbial inoculum Substances 0.000 description 13
- 239000002689 soil Substances 0.000 description 10
- 241001660189 Lysobacter antibioticus Species 0.000 description 9
- 230000002265 prevention Effects 0.000 description 9
- 239000000243 solution Substances 0.000 description 9
- 238000011161 development Methods 0.000 description 7
- 230000018109 developmental process Effects 0.000 description 7
- ODINCKMPIJJUCX-UHFFFAOYSA-N Calcium oxide Chemical compound [Ca]=O ODINCKMPIJJUCX-UHFFFAOYSA-N 0.000 description 6
- 206010028980 Neoplasm Diseases 0.000 description 6
- 241001646398 Pseudomonas chlororaphis Species 0.000 description 6
- 208000015181 infectious disease Diseases 0.000 description 6
- 230000002401 inhibitory effect Effects 0.000 description 6
- 238000004519 manufacturing process Methods 0.000 description 6
- 239000000575 pesticide Substances 0.000 description 6
- 230000000844 anti-bacterial effect Effects 0.000 description 5
- 230000003385 bacteriostatic effect Effects 0.000 description 5
- 239000013043 chemical agent Substances 0.000 description 5
- 230000006806 disease prevention Effects 0.000 description 5
- 238000002474 experimental method Methods 0.000 description 5
- 230000012010 growth Effects 0.000 description 5
- 238000011835 investigation Methods 0.000 description 5
- 238000011160 research Methods 0.000 description 5
- 238000004458 analytical method Methods 0.000 description 4
- 230000003042 antagnostic effect Effects 0.000 description 4
- 230000008901 benefit Effects 0.000 description 4
- 239000012620 biological material Substances 0.000 description 4
- 239000002207 metabolite Substances 0.000 description 4
- 239000000126 substance Substances 0.000 description 4
- 239000005754 Cyazofamid Substances 0.000 description 3
- 206010059866 Drug resistance Diseases 0.000 description 3
- 241000588701 Pectobacterium carotovorum Species 0.000 description 3
- 241000233616 Phytophthora capsici Species 0.000 description 3
- 241001503436 Plasmodiophora brassicae Species 0.000 description 3
- 241000589771 Ralstonia solanacearum Species 0.000 description 3
- 238000012271 agricultural production Methods 0.000 description 3
- 230000009286 beneficial effect Effects 0.000 description 3
- 239000012681 biocontrol agent Substances 0.000 description 3
- 239000000292 calcium oxide Substances 0.000 description 3
- 235000012255 calcium oxide Nutrition 0.000 description 3
- YXKMMRDKEKCERS-UHFFFAOYSA-N cyazofamid Chemical compound CN(C)S(=O)(=O)N1C(C#N)=NC(Cl)=C1C1=CC=C(C)C=C1 YXKMMRDKEKCERS-UHFFFAOYSA-N 0.000 description 3
- 239000003640 drug residue Substances 0.000 description 3
- 244000053095 fungal pathogen Species 0.000 description 3
- 230000036541 health Effects 0.000 description 3
- 238000009630 liquid culture Methods 0.000 description 3
- 230000003071 parasitic effect Effects 0.000 description 3
- 230000002829 reductive effect Effects 0.000 description 3
- 230000004083 survival effect Effects 0.000 description 3
- 238000010998 test method Methods 0.000 description 3
- HSINOMROUCMIEA-FGVHQWLLSA-N (2s,4r)-4-[(3r,5s,6r,7r,8s,9s,10s,13r,14s,17r)-6-ethyl-3,7-dihydroxy-10,13-dimethyl-2,3,4,5,6,7,8,9,11,12,14,15,16,17-tetradecahydro-1h-cyclopenta[a]phenanthren-17-yl]-2-methylpentanoic acid Chemical compound C([C@@]12C)C[C@@H](O)C[C@H]1[C@@H](CC)[C@@H](O)[C@@H]1[C@@H]2CC[C@]2(C)[C@@H]([C@H](C)C[C@H](C)C(O)=O)CC[C@H]21 HSINOMROUCMIEA-FGVHQWLLSA-N 0.000 description 2
- 108010006533 ATP-Binding Cassette Transporters Proteins 0.000 description 2
- 102000005416 ATP-Binding Cassette Transporters Human genes 0.000 description 2
- 241000233866 Fungi Species 0.000 description 2
- 241000223221 Fusarium oxysporum Species 0.000 description 2
- 241000427940 Fusarium solani Species 0.000 description 2
- 241000531155 Pectobacterium Species 0.000 description 2
- 241001503464 Plasmodiophora Species 0.000 description 2
- 240000000851 Vaccinium corymbosum Species 0.000 description 2
- 235000003095 Vaccinium corymbosum Nutrition 0.000 description 2
- 235000017537 Vaccinium myrtillus Nutrition 0.000 description 2
- 238000009825 accumulation Methods 0.000 description 2
- 230000004913 activation Effects 0.000 description 2
- 230000037354 amino acid metabolism Effects 0.000 description 2
- 230000003321 amplification Effects 0.000 description 2
- 239000003899 bactericide agent Substances 0.000 description 2
- 239000003613 bile acid Substances 0.000 description 2
- 230000015572 biosynthetic process Effects 0.000 description 2
- 235000021014 blueberries Nutrition 0.000 description 2
- 238000004364 calculation method Methods 0.000 description 2
- 239000003795 chemical substances by application Substances 0.000 description 2
- 238000010201 enrichment analysis Methods 0.000 description 2
- 230000007613 environmental effect Effects 0.000 description 2
- 238000003912 environmental pollution Methods 0.000 description 2
- 230000001976 improved effect Effects 0.000 description 2
- 230000006872 improvement Effects 0.000 description 2
- 238000000338 in vitro Methods 0.000 description 2
- 230000006799 invasive growth in response to glucose limitation Effects 0.000 description 2
- 238000009629 microbiological culture Methods 0.000 description 2
- 238000005065 mining Methods 0.000 description 2
- 238000003199 nucleic acid amplification method Methods 0.000 description 2
- 230000037361 pathway Effects 0.000 description 2
- 238000012827 research and development Methods 0.000 description 2
- 238000005406 washing Methods 0.000 description 2
- MVXMNHYVCLMLDD-UHFFFAOYSA-N 4-methoxynaphthalene-1-carbaldehyde Chemical compound C1=CC=C2C(OC)=CC=C(C=O)C2=C1 MVXMNHYVCLMLDD-UHFFFAOYSA-N 0.000 description 1
- 241000186361 Actinobacteria <class> Species 0.000 description 1
- 241000589155 Agrobacterium tumefaciens Species 0.000 description 1
- 244000000626 Daucus carota Species 0.000 description 1
- 235000002767 Daucus carota Nutrition 0.000 description 1
- 208000035240 Disease Resistance Diseases 0.000 description 1
- 238000003794 Gram staining Methods 0.000 description 1
- 244000061176 Nicotiana tabacum Species 0.000 description 1
- 235000002637 Nicotiana tabacum Nutrition 0.000 description 1
- 235000014443 Pyrus communis Nutrition 0.000 description 1
- 240000004808 Saccharomyces cerevisiae Species 0.000 description 1
- 241001655322 Streptomycetales Species 0.000 description 1
- 229930006000 Sucrose Natural products 0.000 description 1
- CZMRCDWAGMRECN-UGDNZRGBSA-N Sucrose Chemical compound O[C@H]1[C@H](O)[C@@H](CO)O[C@@]1(CO)O[C@@H]1[C@H](O)[C@@H](O)[C@H](O)[C@@H](CO)O1 CZMRCDWAGMRECN-UGDNZRGBSA-N 0.000 description 1
- 230000005540 biological transmission Effects 0.000 description 1
- 230000000853 biopesticidal effect Effects 0.000 description 1
- MYFXBBAEXORJNB-UHFFFAOYSA-N calcium cyanamide Chemical compound [Ca+2].[N-]=C=[N-] MYFXBBAEXORJNB-UHFFFAOYSA-N 0.000 description 1
- 201000011510 cancer Diseases 0.000 description 1
- 210000004027 cell Anatomy 0.000 description 1
- 210000002421 cell wall Anatomy 0.000 description 1
- 238000003501 co-culture Methods 0.000 description 1
- 239000011248 coating agent Substances 0.000 description 1
- 238000000576 coating method Methods 0.000 description 1
- 230000002860 competitive effect Effects 0.000 description 1
- 238000005138 cryopreservation Methods 0.000 description 1
- 238000012136 culture method Methods 0.000 description 1
- 230000007547 defect Effects 0.000 description 1
- 230000000593 degrading effect Effects 0.000 description 1
- 238000010586 diagram Methods 0.000 description 1
- 238000010790 dilution Methods 0.000 description 1
- 239000012895 dilution Substances 0.000 description 1
- 230000002708 enhancing effect Effects 0.000 description 1
- 210000003608 fece Anatomy 0.000 description 1
- 230000035558 fertility Effects 0.000 description 1
- 239000003337 fertilizer Substances 0.000 description 1
- 229940107131 ginseng root Drugs 0.000 description 1
- 230000009422 growth inhibiting effect Effects 0.000 description 1
- 230000001939 inductive effect Effects 0.000 description 1
- 238000009776 industrial production Methods 0.000 description 1
- 230000005764 inhibitory process Effects 0.000 description 1
- 239000003621 irrigation water Substances 0.000 description 1
- 230000002147 killing effect Effects 0.000 description 1
- 230000000670 limiting effect Effects 0.000 description 1
- 244000144972 livestock Species 0.000 description 1
- 239000010871 livestock manure Substances 0.000 description 1
- 230000007774 longterm Effects 0.000 description 1
- 238000005259 measurement Methods 0.000 description 1
- 230000007246 mechanism Effects 0.000 description 1
- 230000000813 microbial effect Effects 0.000 description 1
- 230000002906 microbiologic effect Effects 0.000 description 1
- 238000002156 mixing Methods 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
- 230000000877 morphologic effect Effects 0.000 description 1
- 235000015097 nutrients Nutrition 0.000 description 1
- 235000003715 nutritional status Nutrition 0.000 description 1
- 244000052769 pathogen Species 0.000 description 1
- 230000001717 pathogenic effect Effects 0.000 description 1
- 230000008635 plant growth Effects 0.000 description 1
- 238000007747 plating Methods 0.000 description 1
- 230000008092 positive effect Effects 0.000 description 1
- 230000001737 promoting effect Effects 0.000 description 1
- 229930000044 secondary metabolite Natural products 0.000 description 1
- 238000000926 separation method Methods 0.000 description 1
- 239000007787 solid Substances 0.000 description 1
- 230000004763 spore germination Effects 0.000 description 1
- 238000003860 storage Methods 0.000 description 1
- 239000005720 sucrose Substances 0.000 description 1
- 208000024891 symptom Diseases 0.000 description 1
- 230000009885 systemic effect Effects 0.000 description 1
- 230000035899 viability Effects 0.000 description 1
Images
Classifications
-
- 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
- A01N25/00—Biocides, pest repellants or attractants, or plant growth regulators, characterised by their forms, or by their non-active ingredients or by their methods of application, e.g. seed treatment or sequential application; Substances for reducing the noxious effect of the active ingredients to organisms other than pests
- A01N25/02—Biocides, pest repellants or attractants, or plant growth regulators, characterised by their forms, or by their non-active ingredients or by their methods of application, e.g. seed treatment or sequential application; Substances for reducing the noxious effect of the active ingredients to organisms other than pests containing liquids as carriers, diluents or solvents
-
- 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
- A01N63/20—Bacteria; Substances produced thereby or obtained therefrom
- A01N63/25—Paenibacillus
-
- 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)
- Health & Medical Sciences (AREA)
- Engineering & Computer Science (AREA)
- General Health & Medical Sciences (AREA)
- Zoology (AREA)
- Wood Science & Technology (AREA)
- Chemical & Material Sciences (AREA)
- Biotechnology (AREA)
- Agronomy & Crop Science (AREA)
- Organic Chemistry (AREA)
- Genetics & Genomics (AREA)
- Microbiology (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Virology (AREA)
- Environmental Sciences (AREA)
- Dentistry (AREA)
- Plant Pathology (AREA)
- Pest Control & Pesticides (AREA)
- Medicinal Chemistry (AREA)
- Biomedical Technology (AREA)
- Tropical Medicine & Parasitology (AREA)
- General Engineering & Computer Science (AREA)
- Biochemistry (AREA)
- Toxicology (AREA)
- Agricultural Chemicals And Associated Chemicals (AREA)
Abstract
The invention belongs to the technical field of microorganisms, and discloses a paenibacillus polymyxa strain and application thereof, wherein the paenibacillus polymyxa strain P21 is registered in the general microorganism center of the China Committee for culture Collection of microorganisms at 12 months and 6 days 2021, and the preservation numbers are as follows: CGMCC No. 24044; the paenibacillus polymyxa strain P21 can be applied to the preparation of water aqua for preventing and treating clubroot of cruciferous vegetables and root rot of panax notoginseng; the preparation method of the paenibacillus polymyxa strain P21 aqueous solution comprises the following steps: strain culture, including test tube strain culture and strain shaking table enlarged culture; fermentation, including preparation of seed suspensions, inoculation, and liquid fermentation. The paenibacillus polymyxa P21 has good comprehensive properties, can be used for preventing and treating clubroot and panax notoginseng root rot of cruciferous vegetables (such as Chinese cabbages, baby cabbages and the like), and is safe to use without residues.
Description
Technical Field
The invention belongs to the technical field of microorganisms, and particularly relates to a paenibacillus polymyxa strain and application thereof.
Background
At present, clubroot is a soil-borne disease which has a large influence in world agricultural production and mainly harms cruciferous vegetables and rapes to cause underground root tumors, the overground part of the plant withers and finally the plant dies. Is a worldwide disease caused by the infection of Plasmodiophora brassicae. Generally, the seedling stage is damaged, and in severe cases, seedlings wither, the yield is seriously reduced, and even the seedlings are not harvested; once the field is infected by plasmodiophora brassicae, the soil can carry bacteria for a long time, and the sustainable production of cruciferous crops is seriously threatened. In recent years, the incidence of clubroot becomes more serious worldwide, especially in regions such as Asia, northern Europe, Australia and North America, clubroot becomes a main disease of cruciferous crops, with the continuous improvement of mechanization degree and the popularization of greenhouse seedling and floating seedling, the circulation speed and the damage range of clubroot diseases are further accelerated, particularly in Yunnan and Sichuan regions, the clubroot diseases in main rape producing areas are more serious, the area of land infected by the clubroot diseases of some countries is close to 100%, and the economic loss is even more than 60%.
Plasmodiophora is an obligate parasitic bacterium, and the germ survives in winter in soil or plasmodiophora tissues as dormant spores. The clubroot infection cycle is divided into three stages of survival, root hair infection and cortex infection in dormant spore soil. The production mainly comprises the steps of killing dormant spores or inhibiting the dormant spores from sprouting, for example, applying bactericides or biocontrol bactericides to soil to reduce root hair infection or cortex infection and relieve clubroot. Clubroot transmission is mainly transmitted through soil, agricultural implements, irrigation water, rainwater and manure carrying dormant spores. The clubroot is extremely difficult to prevent and treat due to the fact that the clubroot belongs to obligate parasitic bacteria, and the traditional prevention and treatment idea is to apply quicklime or lime nitrogen (calcium cyanamide) in soil, improve the pH value of the soil and reduce the occurrence of the clubroot. However, long-term application or excessive application of quicklime cannot achieve good disease control effect in many cases except for damage to soil structure. The biopesticide has various advantages, has the characteristics of wide sources, difficult generation of drug resistance, safety to people and livestock, good environmental compatibility, multiple development and utilization ways, various varieties, large research and development choices and the like, provides a very important research direction for agricultural sustainable development, and has become a new trend of the global agricultural production development.
The application of chemical agents to control crop soil-borne diseases plays an important role in agricultural production, however, the problems of drug residue, environmental pollution, drug resistance accumulation and the like caused by the large-scale use of chemical agents do not meet the requirements of sustainable development of agricultural health, biological control becomes a research hotspot for controlling plant soil-borne diseases at home and abroad due to the characteristics of low cost, environmental friendliness, no drug residue and the like, the traditional chemical control means is gradually replaced, and the application prospect is wide. Biological control refers to a method of using one or more microorganisms to inhibit the viability and fertility of a pathogen. Common mechanisms for biological control of plant diseases are: improving the physical and chemical properties and the nutritional status of soil, promoting the growth of plants, improving the health level of the plants and enhancing the disease resistance of host plants; the parasitic and antagonistic actions of antagonistic microorganisms such as biocontrol bacteria, streptomycete, actinomycetes and the like and the competitive effects of antagonistic microorganisms and nutrient substances and ecological niches of pathogenic bacteria are utilized to inhibit and eliminate the pathogenic bacteria; inducing the host plant to develop systemic resistance to the pathogenic bacteria.
Early stage research bases of metabonomics and cultigenmics show that the paenibacillus polymyxa strain P21 has good compatibility with other strains (lysobacter antibioticus, pseudomonas chlororaphis and the like), and greenhouse and field experiments prove that the relative control effect of the combination of the paenibacillus polymyxa and the lysobacter antibioticus is obviously improved compared with that of single biocontrol bacteria; the enrichment analysis of the KEGG pathway shows that the differential metabolite mainly participates in amino acid metabolism, bile acid biosynthesis, ABC transporter, and the variety and content of the metabolite play important roles in the biological prevention and treatment of the clubroot of the Chinese cabbage. And the paenibacillus polymyxa strain P21 has good colonization characteristics in the tissue of the Chinese cabbage root, has strong adaptability to the ecological environment, and is more beneficial to the prevention and treatment of the vegetable clubroot. Therefore, the paenibacillus polymyxa strain P21 has good mining potential and development prospect.
Through the above analysis, the problems and defects of the prior art are as follows:
(1) in the existing method for preventing and controlling plasmodiophora brassicae, quicklime is applied for a long time or excessively, and besides damage to a soil structure, a good disease control effect cannot be achieved in many cases.
(2) In the existing method for preventing and treating soil-borne diseases of crops by applying chemical agents, the problems of drug residue, environmental pollution, drug resistance accumulation and the like caused by the large-scale use of the chemical agents do not meet the sustainable development requirement of agricultural health.
Disclosure of Invention
Aiming at the problems in the prior art, the invention provides a Paenibacillus polymyxa strain P21 and application thereof.
The invention is realized by the following steps that a paenibacillus polymyxa strain P21 is registered in the general microorganism center of China Committee for culture Collection of microorganisms No. 12/6 2021, wherein the preservation numbers are as follows: CGMCC No. 24044.
The invention also aims to provide application of the paenibacillus polymyxa strain P21 in preparing a water aqua for preventing and treating clubroot of cruciferous vegetables and root rot of panax notoginseng.
Further, the preparation method of the paenibacillus polymyxa strain P21 aqueous solution comprises the following steps:
step one, strain culture, including test tube strain culture and strain table expansion culture;
and step two, fermentation, which comprises the preparation of seed suspension, inoculation and liquid fermentation.
Further, the tube seed culture in the first step comprises:
sterilizing NA culture medium at 121 deg.C for 30min, pouring into 9cm plate to obtain NA culture substrate plate, inoculating Paenibacillus polymyxa P21, and culturing at 28 deg.C for 36 h.
Further, the formula of the NA culture medium is as follows: 10.0g of glucose, 5.0g of peptone, 3.0g of beef extract, 1.0g of yeast extract, 17.0-20.0 g of agar powder and 1000mL of distilled water.
Further, the strain shaking table expanded culture in the step one comprises the following steps:
inoculating the flat seeds on an eggplant bottle slant NA culture medium, and culturing at 28 ℃ for 36h to obtain eggplant bottle seeds.
Further, the preparation of the seed suspension in the second step comprises:
the colonies on the slant of the eggplant bottle were washed with sterilized water to obtain a seed suspension.
Further, the inoculation in the second step comprises:
the seed suspension is inoculated into the culture medium of the fermentation tank by the pressure difference through an inoculation port.
The formula and the processing method of the fermentation tank culture medium are as follows: peptone 0.5%, glucose 0.7%, yeast extract 0.1%, magnesium sulfate 0.15%, dipotassium hydrogen phosphate 0.15%, glycerol 10%, distilled water 1000mL, pH 7.5 before sterilization.
Further, the liquid fermentation in the second step comprises:
after inoculation, culturing the fermentation tank culture medium for 36-48 h; wherein the temperature of the fermentation tank is 26-32 ℃, and the pressure of the fermentation tank is 0.5kg/cm 2 The stirring speed in the tank is 180 rpm; in the process of inoculation and pressure increase, the tank pressure does not exceed 1.5kg/cm 2 。
Further, the application method of the paenibacillus polymyxa strain P21 aqueous solution is root irrigation.
In combination with the technical solutions and the technical problems to be solved, please analyze the advantages and positive effects of the technical solutions to be protected in the present invention from the following aspects:
first, aiming at the technical problems existing in the prior art and the difficulty in solving the problems, the technical problems to be solved by the technical scheme of the present invention are closely combined with results, data and the like in the research and development process, and some creative technical effects are brought after the problems are solved. The specific description is as follows:
the paenibacillus polymyxa strain P21 provided by the invention is separated from healthy Chinese cabbage root tissues in Tonghai county of Yuxi city, Yunnan, and the paenibacillus polymyxa strain P21 can be prepared into an aqueous solution, can act on vegetables (Chinese cabbage, baby cabbage and the like) and pseudo-ginseng economic crops, and is used for preventing and treating vegetable clubroot and pseudo-ginseng root rot.
The paenibacillus polymyxa strain P21 provided by the invention can be prepared into an aqueous solution, acts on crops such as baby cabbage, Chinese cabbage, pseudo-ginseng and the like, and generates secondary metabolites: antibacterial related substances such as fengyuan and iturin are used for degrading cell walls of pathogenic bacteria, inhibiting spore germination and sprout tube elongation of the pathogenic bacteria, and preventing and treating clubroot of cruciferous vegetables and root rot of panax notoginseng. The water aqua is safe to use without residue, is beneficial to reducing the use amount and the residual amount of chemical agents in agricultural products, reduces the production cost, has strong practicability, and is suitable for industrial production.
Secondly, considering the technical scheme as a whole or from the perspective of products, the technical effect and advantages of the technical scheme to be protected by the invention are specifically described as follows:
the Paenibacillus polymyxa P21 provided by the invention has good comprehensive properties, and can be used for preventing and treating clubroot and root rot of panax notoginseng of cruciferous vegetables (such as Chinese cabbage, baby cabbage and the like).
Third, as an inventive supplementary proof of the claims of the present invention, there are also presented several important aspects:
the paenibacillus polymyxa strain P21 is obtained by separating healthy Chinese cabbage root tissues from Tonghai county of Yuxi city, Yunnan, and the paenibacillus polymyxa strain P21 is deposited in the general microbiological center registration book of China Committee for culture Collection of microorganisms at 12 months and 6 days 2021, with the preservation number of CGMCC No. 24044. The paenibacillus polymyxa strain P21 can be applied to the preparation of water aqua for preventing and treating clubroot of cruciferous vegetables and root rot of panax notoginseng; the preparation method of the paenibacillus polymyxa strain P21 aqueous solution comprises the following steps: strain culture, including test tube strain culture and strain shaking table enlarged culture; fermentation, including preparation of seed suspensions, inoculation, and liquid fermentation. The Paenibacillus polymyxa P21 has good comprehensive properties, can be used for preventing and treating clubroot and root rot of panax notoginseng of cruciferous vegetables (such as Chinese cabbages, baby cabbages and the like), and is safe to use without residues.
Early-stage research bases of metabonomics and cultivamics show that the paenibacillus polymyxa strain P21 can have good compatibility with other strains (lysobacter antibioticus, pseudomonas chlororaphis and the like), and greenhouse and field experiments prove that the relative control effect is remarkably improved compared with single biocontrol bacteria by the combination of the paenibacillus polymyxa and the lysobacter antibioticus; the enrichment analysis of the KEGG pathway shows that the differential metabolite mainly participates in amino acid metabolism, bile acid biosynthesis, ABC transporter, and the variety and content of the metabolite play important roles in the biological prevention and treatment of the clubroot of the Chinese cabbage.
The expected income and commercial value after the technical scheme of the invention is that the paenibacillus polymyxa strain P21 is easy to culture and separate, can have good colonization characteristic in the root tissue of the Chinese cabbage, has strong adaptability to the ecological environment, and is more beneficial to the prevention and treatment of the clubroot of vegetables. Therefore, the paenibacillus polymyxa strain P21 has good mining potential and development prospect.
Drawings
In order to more clearly illustrate the technical solutions of the embodiments of the present invention, the drawings needed to be used in the embodiments of the present invention will be briefly described below, and it is obvious that the drawings described below are only some embodiments of the present invention, and it is obvious for those skilled in the art that other drawings can be obtained according to the drawings without creative efforts.
FIG. 1 is a flow chart of a method for preparing an aqueous solution of Paenibacillus polymyxa strain P21 according to an embodiment of the present invention;
FIG. 2 is a colony morphology diagram of a Paenibacillus polymyxa strain (Paenibacillus polymyxa) P21 provided in the examples of the present invention; in the figure: a is a streak chart of pure culture of P21; b is a single colony image under plate-coating counting.
Detailed Description
In order to make the objects, technical solutions and advantages of the present invention more apparent, the present invention is further described in detail with reference to the following embodiments. It should be understood that the specific embodiments described herein are merely illustrative of the invention and are not intended to limit the invention.
Aiming at the problems in the prior art, the invention provides a paenibacillus polymyxa strain and application thereof, and the invention is described in detail with reference to the accompanying drawings.
This section is an explanatory embodiment expanding on the claims so as to fully understand how the present invention is embodied by those skilled in the art.
The paenibacillus polymyxa strain P21 provided by the embodiment of the invention is classified and named as paenibacillus polymyxa, and the paenibacillus polymyxa strain P21 is registered in the general microbiological culture collection center of China Committee for culture Collection of microorganisms at 12 months and 6 days 2021, wherein the preservation numbers are as follows: CGMCC No. 24044. China general microbiological culture Collection center, addresses of West Lu No. 1 Hospital No. 3 of the sunward region of Beijing, institute of microbiology of China academy of sciences, postal code 100101, telephone 010 plus 64807355, fax 010 plus 64807288, electronic mail cgmcc @ im. ac. cn htp:// w cgmcc net; the biological material of the references, P21; the classification nomenclature proposed in (1) is Paenibailluspolymyxa; this biological material (strain) was received from the depository at 12.06.2021 and was registered in the book. Storage is for thirty years from that day on request and continues for five years after a request to provide a sample of biological material is received before expiration. The survival of the biological material was examined at 12/06/2021 by the Collection, and the result was survival.
The application of the paenibacillus polymyxa strain P21 provided by the embodiment of the invention is as follows: the Bacillus polymyxa strain P21 is prepared into aqua for preventing and treating clubroot and Notoginseng radix root rot of cruciferous vegetables (such as baby cabbage and Chinese cabbage).
As shown in fig. 1, the preparation method of the paenibacillus polymyxa strain P21 aqueous solution provided by the embodiment of the invention comprises the following steps:
s101, strain culture, including test tube strain culture and strain shaking table expansion culture;
and S102, fermenting, wherein the fermentation comprises the preparation of seed suspension, inoculation and liquid fermentation.
The paenibacillus polymyxa P21 provided by the embodiment of the invention is prepared into an aqueous solution, and the preparation method comprises the following steps:
(1) strain culture
a. And (3) test tube seed culture: sterilizing NA culture medium at 121 deg.C for 30min, pouring into 9cm plate to obtain NA culture substrate plate, inoculating Paenibacillus polymyxa P21, and culturing at 28 deg.C for 36 hr;
the formula of the NA culture medium is as follows: 10.0g of glucose, 5.0g of peptone, 3.0g of beef extract, 1.0g of yeast extract, 17.0-20.0 g of agar powder and 1000mL of distilled water;
b. and (3) expanding and culturing a strain by using a shaking table: inoculating the flat seeds to an eggplant bottle slant NA culture medium, and culturing at 28 ℃ for 24-48 h to obtain eggplant bottle seeds;
(2) fermentation of
a. Preparation of seed suspension: washing the bacterial colony on the slope of the eggplant bottle by using sterilized water to obtain a seed suspension;
b. inoculation: the seed suspension is inoculated into a fermentation tank culture medium by differential pressure through an inoculation port;
the formula and the processing method of the fermentation tank culture medium are as follows: peptone 0.5%, glucose 0.7%, yeast extract 0.1%, magnesium sulfate 0.15%, dipotassium hydrogen phosphate 0.15%, glycerol 10%, distilled water 1000mL, pH 7.5 before sterilization;
c. liquid fermentation: and after inoculation, culturing the fermentation tank culture medium for 36-48 h. The temperature of the fermentation tank is 26-32 ℃, and the pressure of the fermentation tank is 0.5kg/cm 2 The stirring speed in the tank was 180 rpm. In the process of inoculation and pressure increase, the tank pressure does not exceed 1.5kg/cm 2 。
The application method of the paenibacillus polymyxa P21 aqueous solution provided by the embodiment of the invention is root irrigation.
The paenibacillus polymyxa P21 strain has the following morphological characteristics:
the P21 colony of Paenibacillus polymyxa strain (Paenibacillus polymyxa) is shown in FIG. 2, and is white, round, small, smooth, moist, and sticky in surface. Gram staining is positive, the thallus is rod-shaped and has spores, and the optimal growth temperature is 28 ℃.
Example 1: and (3) preparing a paenibacillus polymyxa strain P21 aqueous solution.
(1) Strain culture
a. Test tube culture (the following are in weight percent)
Putting an NA culture medium (5.0 g of peptone, 10.0g of sucrose, 1.0g of yeast powder, 3.0g of beef extract and 17.0-20.0 g of agar) into a test tube, sterilizing with 1000mL of distilled water, making a flat plate, inoculating Paenibacillus polymyxa strain P21, and culturing at 28 ℃ for 36 h.
b. Enlarged culture of strain by shaking bed
Inoculating the flat seeds on an eggplant bottle slant NA culture medium (the culture medium formula is the same as the step a) and culturing for 48h at 26 ℃ to obtain eggplant bottle seeds.
(2) Fermentation of
The specific fermentation production process is as follows:
a. preparation of seed suspension: washing the bacterial colony on the slope of the eggplant bottle by using sterilized water to obtain a seed suspension;
b. inoculation: inoculating the seed suspension into a fermentation tank (culture medium of the fermentation tank comprises peptone 0.5%, glucose 0.7%, yeast extract 0.1%, magnesium sulfate 0.15%, dipotassium hydrogen phosphate 0.15%, glycerol 10%, pH 8.0 before sterilization, temperature of the fermentation tank is 26-32 deg.C, and tank pressure is 0.5kg/cm 2 Stirring speed in the tank is 180rpm), and attention is paid to avoid pollution; the pot pressure can not exceed 1.5kg/cm during pressure increase 2 ；
c. Liquid fermentation:
after inoculation, culturing the fermentation tank for 36h until the number of bacteria is not increased any more, immediately placing the fermentation tank, and measuring the bacteria content through plate counting, wherein the bacteria content reaches 15 hundred million CFU/mL;
the preparation can be applied to roots of crops in the early growth stage and the disease stage in a root irrigation mode to serve as a disease prevention and control and yield increase measure, and the method comprises the following steps: the biological preparation is adopted to irrigate roots of crops to prevent and control diseases in the early growth period or the initial disease period of the crops. The fertilizer can be applied 3 times in the growing process of crops in severe areas of the crops.
Example 2: coculture compatibility test of Paenibacillus polymyxa (Paenibacillus polymyxa) P21 and Lysobacter antibioticus (Lysobacter antibioticus) 13-6.
The media used were as follows:
KB Medium, R 2 And (A) culture medium.
The test method comprises the following steps:
two biocontrol strains, one of which is Lysobacter antibioticus (Lysobacter antibioticus)13-6 (disclosed in the patent specification with the application number of ZL 200910094763.6) and the other of which is Paenibacillus polymyxa P21, are selected and subjected to single culture and co-culture respectively: (1) p21; (2) 13-6; (3)13-6+ P21. Culturing each strain of bacteria in KB culture medium for 48h, picking out single colony from each pure culture medium, transferring into 50mL KB liquid culture medium, and culturing at 28 ℃ and 150r/min for 24h to obtain seed liquid. Inoculating 13-6 seed solution at a ratio of 1% into 500mL KB liquid culture medium, culturing at 28 deg.C and 150r/min for 48h, inoculating P21 seed solution at a ratio of 1%, and culturing at 28 deg.C and 150r/min for 12 h. Diluted single and co-cultured bacterial culture (10) -7 ) 0.1mL of each dilution was pipetted and applied to R 2 And A, counting according to different colony morphologies.
The colony number of each biocontrol bacterium is obviously increased in the process of co-culturing two biocontrol bacteria in the same concentration plating separation, which shows that the Paenibacillus polymyxa P21 and the Lysobacter antibioticus 13-6 have good compatibility, and the two biocontrol bacteria can promote the growth of other strains.
Example 3: in vitro bacteriostatic test of Paenibacillus polymyxa (Paenibacillus polymyxa) P21 against different pathogenic bacteria.
The media used were as follows:
YEM medium, NA medium, PDA medium.
In-vitro bacteriostasis test of the bacterial agent of Paenibacillus polymyxa P21 on pectobacterium carotovorum and other pathogenic bacteria.
Pathogenic bacteria to be tested:
including carrot soft rot Pectobacterium (Pectibacter carotovorum), Fusarium oxysporum (Fusarium oxysporum), Phytophthora capsici (Phytophtora capsici), Fusarium solani (Fusarium solani), tobacco bacterial wilt (Ralstonia solanacearum), blueberry crown gall (Agrobacterium tumefaciens) and the like, all of which are provided by the bacterial research laboratory of Yunnan agricultural university.
The test method comprises the following steps:
activation of pathogenic fungi: transferring the stored pathogenic bacteria strain into a PDA culture medium, performing amplification culture at a constant temperature of 26 ℃, and performing a bacteriostatic experiment for later use when hyphae grow over the culture dish.
Activation of bacteria: the method comprises the steps of sucking 80 mu L of bacterial liquid from a bacterial cryopreservation tube stored at the temperature of-80 ℃ into an NA culture medium by using Pectobacterium carotovorum, Ralstonia solanacearum and blueberry root cancer bacteria. All tested pathogenic bacteria are subjected to inverted culture at 28 ℃ for 48h, after the bacteria are activated, a single colony is selected for streak amplification culture, and is subjected to inverted culture at 28 ℃ for 48h for standby of a bacteriostatic experiment.
The determination method comprises the following steps:
and (3) measuring the bacteriostatic rate of the pathogenic fungi: the inhibition capacity of each strain on the growth of pathogenic bacteria is detected by adopting a plate confronting culture method, in a superclean bench, pathogenic bacteria growing on PDA are made into a bacterial cake with the diameter of 5mm by using a puncher, then the bacterial cake is transferred to the center of a PDA culture medium plate, separated and purified biocontrol bacteria P21 are connected to a position 2.5cm away from the pathogenic bacteria by using a liquid-transferring gun head, each dish is inoculated with a strain of test bacteria, 3 times of repetition are arranged, meanwhile, the plate only containing the pathogenic bacteria cake is used as a contrast, a fungus constant-temperature incubator at 26 ℃ is used for culture, and the width of an antibacterial belt is measured when CK grows to be full of the culture medium.
Measurement of the bacteriostatic rate of pathogenic bacteria: selecting a single colony of activated and pathogenic bacteria, inoculating the single colony into a 100mLNA liquid culture medium, performing shaking culture at 28 ℃ and 160rpm for 36 hours to prepare a bacterial suspension, uniformly mixing the bacterial suspension with an NA solid culture medium at 45 ℃ to prepare a bacterial carrying bottom plate, inoculating biocontrol bacteria P21 to the center of the bacterial carrying bottom plate on an ultraclean workbench, performing inverted culture at 28 ℃ for 48 hours, and measuring the width of the antibacterial band.
Test results and analysis:
the test results are shown in Table 1.
TABLE 1 determination of the growth inhibitory effect of Paenibacillus polymyxa P21 on different pathogenic bacteria
As can be seen from table 1, Paenibacillus polymyxa P21 has strong antagonistic action against 8 pathogenic bacteria and fungi to be tested, wherein Paenibacillus polymyxa P21 has the strongest inhibitory action against Phytophthora capsici (Phytophthora capsici), Paenibacillus polymyxa P21 has the stronger inhibitory action against Pectobacterium carotovorum (Pectobacterium), but has weak inhibitory action against Ralstonia solanacearum. And the paenibacillus polymyxa P21 has better inhibitory effect on pathogenic fungi than on pathogenic bacteria.
Example 4: and (3) field prevention and control test of the paenibacillus polymyxa strain P21 on the clubroot of baby cabbage.
Test work: the baby cabbage variety (golden silk white) is susceptible to clubroot.
Test medium: KB liquid medium
The experimental site: vegetable test field in Shanghai county and Jia pear garden in Yuxi city, Yunnan province
Preparing a biocontrol microbial inoculum:
inoculating Paenibacillus polymyxa strain P21 into a triangular flask containing 500mL KB liquid medium, performing shake culture in a shaking table (28 ℃, 160rpm) for 3d, and adjusting OD of bacterial suspension by using an ultraviolet spectrophotometer 600nm The concentration of the bacterial suspension was equivalent to 1X 10 ═ 0.5 8 CFU/mL for use.
The experimental method comprises the following steps:
the method comprises the steps of setting 3 treatments of a biocontrol microbial inoculum treatment, a clear water control and a pesticide control in a test, adopting a root irrigation method for the pesticide application, irrigating 300mL of biocontrol microbial inoculum at the root of each Chinese cabbage, applying the biocontrol microbial inoculum for the first time in the initial disease stage, applying the biocontrol microbial inoculum for 3 times after 7 days, carrying out disease condition investigation after 50 days, recording the disease incidence and disease condition index of the clubroot of the baby cabbage, calculating the control effect and analyzing the result. 30 baby cabbage plants are randomly selected for investigation and are repeatedly studied for 3 times, grading is carried out according to the incidence condition of clubroot (see table 2), and the incidence rate, disease index and disease prevention effect of each treatment are calculated according to the following calculation mode.
TABLE 2 grading Standard of clubroot
The disease incidence is the number of diseased plants/total plants multiplied by 100%
Disease index ═ Σ (number of diseased plants at each stage x relative number) × 100/(total number of investigated plants × 9)
The prevention and treatment effect is [ (control disease index-treatment disease index)/control disease index ] × 100%
TABLE 3 determination of field control effect of biocontrol microbial inoculum P21 on cabbage clubroot
As can be seen from Table 3, the biocontrol agent Paenibacillus polymyxa strain P21 applied in the field has obvious control effect on the clubroot of baby cabbage and has control effect of 72.65%. Compared with a control and a pesticide cyazofamid, the disease incidence and the disease index of the applied biocontrol microbial agent paenibacillus polymyxa P21 are obviously reduced, and the prevention effect is most obvious. The biological control agent paenibacillus polymyxa P21 has good control effect on the clubroot of the baby cabbage, the control effect test has repeatability, the biological control agent paenibacillus polymyxa P21 has stable and good control effect on the clubroot of the baby cabbage, and the basic conditions for mass popularization in production are also met.
Example 5: disease prevention test of paenibacillus polymyxa strain P21 on biennial root rot of panax notoginseng in greenhouses.
Test work: radix Notoginseng for two years
Test medium: KB liquid medium
The experimental site: wenshanzhou inkstone county flat far street pseudo-ginseng farmer planting greenhouse
Preparing a biocontrol microbial inoculum:
inoculating Paenibacillus polymyxa strain P21 into a triangular flask containing 500mL KB liquid medium, performing shake culture in a shaking table (28 ℃, 160rpm) for 3d, and adjusting OD of bacterial suspension by using an ultraviolet spectrophotometer 600 The concentration of the bacterial suspension is equivalent to 1X 10 ═ 0.5 8 CFU/mL for use.
The test method comprises the following steps:
the test sets the biocontrol microbial inoculum treatment and the clear water comparison, the root irrigation method is adopted, 500mL of biocontrol bacterial liquid is irrigated at the root of each panax notoginseng two years later, the biocontrol bacterial liquid is applied 3 times after 7 days, the disease condition investigation is carried out after 90 days, the morbidity and the disease condition index of panax notoginseng root rot are recorded, the control effect is calculated, and the result is analyzed. 10 biennial panax notoginseng is randomly selected from each treated three-time repeated cell for investigation, grading is carried out according to the incidence condition of the root rot (see table 2), and the incidence rate, disease index and disease prevention effect of each treatment are carried out according to the following calculation mode.
The field grading standard of the root rot of panax notoginseng: grade 0, healthy, asymptomatic; level 1, the rot only occurs on the surface of the root tuber, and 0< the rot area ratio is less than or equal to 10 percent, or the rot is expanded to the inside and 5% < the rot area ratio is less than or equal to 10 percent; 2, the rot is expanded to the inside, the ratio of the rot area is less than or equal to 40 percent after 10 percent, and the branch fibrous roots are not rotten; grade 3, the rotting is expanded to the inside, the rotting area ratio is less than or equal to 50% after 40%, and the branch fibrous roots are rotted; level 4, the rot is expanded to the inside, 50 percent of the rot area is less than or equal to 70 percent, and the branch fibrous roots are rotten and fall off; grade 5, decay has spread to the inside and the rotted area accounts for > 70% until the whole root tuber is completely rotten and the branch fibrous root is rotten.
The disease incidence is the number of diseased plants/the total number of plants multiplied by 100%
Disease index [ Σ (diseased plant number at each stage × representative value at each stage)/total investigated plant number/highest representative value ] × 100
The prevention and treatment effect is [ (control disease index-treatment disease index)/control disease index ] × 100%
Test results and analysis
The test results are shown in Table 4.
TABLE 4 disease prevention effect of Paenibacillus polymyxa strain P21 on panax notoginseng root rot
As can be seen from table 4, the application of paenibacillus polymyxa P21 has a certain control effect on biennial panax notoginseng root rot, and significantly reduces the incidence and disease index of biennial panax notoginseng root rot.
Example 6: a paenibacillus polymyxa strain (Paenibacillus polymyxa) P21 is used for a field disease control test of Chinese cabbage clubroot.
The test samples and media used in the test were as follows:
KB Medium and NA Medium
Test work: chinese cabbage (Luchunbai No. 1, 83-1) is susceptible to clubroot of cruciferous vegetables;
the control object is: clubroot of Chinese cabbage
The experimental site: large pendulum village Chinese cabbage clubroot test field in Panlongdistrict of Kunming, Yunnan province
Preparing a biocontrol microbial inoculum: biocontrol agents were prepared as described in example 3
The inoculation method comprises the following steps:
the method comprises the steps of setting biocontrol microbial inoculum treatment, clear water control and pesticide cyazofamid 200-time solution control for 3 treatments in the test, adopting a root irrigation method for the pesticide application method, irrigating 300mL of biocontrol bacterial solution at the root of each Chinese cabbage, applying the biocontrol microbial inoculum for the first time in the early stage of disease occurrence, applying the biocontrol bacterial solution for 3 times after 7 days, carrying out disease condition investigation after 50 days, recording the disease incidence and disease condition index of the clubroot disease of the Chinese cabbage, calculating the control effect and analyzing the result.
According to the grading standard of clubroot disease conditions: level 0: no disease symptoms; level 1: tumors were found only on the fibrous roots; and 3, level: tumors were found on lateral roots; and 5, stage: lateral root with large tumor and distal main root with small tumor; and 7, stage: the main roots are all swollen, but the plant grows normally; and 9, stage: the swollen part of the root starts to crack and rot, and the plant withers or dies.
Disease index ═ Σ (number of diseased plants at each stage × relative stage value) × 100/(number of total investigated plants × highest stage value)
Relative control effect (control disease index-treatment disease index)/control disease index x 100%
Test results and analysis
The results are shown in Table 5.
TABLE 5 determination of field control effect of biocontrol microbial inoculum P21 on clubroot disease of Chinese cabbage
As can be seen from Table 5, the biocontrol microbial inoculum Paenibacillus polymyxa P21 applied in the field has obvious control effect on the clubroot of Chinese cabbage, and the control effect is 64.19%. Compared with a control and a pesticide cyazofamid, the incidence rate and the disease index of the Paenibacillus polymyxa strain (Paenibacillus polymyxa) P21 applied with the biocontrol agent are obviously reduced. The biological control agent paenibacillus polymyxa P21 is proved to have good control effect on the clubroot of the cabbage, the control effect test has repeatability, the biological control agent paenibacillus polymyxa P21 is proved to have stable and good control effect on the clubroot of the cabbage, and the basic conditions for mass popularization in production are also met.
The above description is only for the purpose of illustrating the present invention and the appended claims are not to be construed as limiting the scope of the invention, which is intended to cover all modifications, equivalents and improvements that are within the spirit and scope of the invention as defined by the appended claims.
Claims (10)
1. A paenibacillus polymyxa strain P21, wherein the paenibacillus polymyxa strain P21 was registered in the common microorganism center of the china committee for culture collection of microorganisms at 12 months and 6 days 2021, with the collection numbers: CGMCC No. 24044.
2. Use of the paenibacillus polymyxa strain P21 of claim 1 in the preparation of an aqueous solution for preventing and treating clubroot of cruciferous vegetables and root rot of panax notoginseng.
3. The application of the paenibacillus polymyxa strain P21 in preparing an aqueous solution for preventing and treating clubroot disease of cruciferous vegetables and root rot disease of panax notoginseng according to claim 2, wherein the preparation method of the paenibacillus polymyxa strain P21 aqueous solution comprises the following steps:
step one, strain culture, including test tube strain culture and strain shaking table enlarged culture;
and step two, fermentation, which comprises the preparation of seed suspension, inoculation and liquid fermentation.
4. The use of paenibacillus polymyxa strain P21 in the preparation of an aqueous solution for preventing and treating clubroot of cruciferous vegetables and root rot of panax notoginseng according to claim 3, wherein the tube seed culture in the first step comprises:
sterilizing NA culture medium at 121 deg.C for 30min, pouring into 9cm plate to obtain NA culture substrate plate, inoculating Paenibacillus polymyxa P21, and culturing at 28 deg.C for 36 h.
5. The use of Paenibacillus polymyxa strain P21 in the preparation of an aqueous solution for preventing and treating clubroot of cruciferous vegetables and root rot of pseudo-ginseng according to claim 4, wherein the formula of the NA medium is as follows: 10.0g of glucose, 5.0g of peptone, 3.0g of beef extract, 1.0g of yeast extract, 17.0-20.0 g of agar powder and 1000mL of distilled water.
6. The use of Paenibacillus polymyxa strain P21 in the preparation of an aqueous solution for preventing and treating clubroot of cruciferous vegetables and root rot of Panax notoginseng as claimed in claim 3, wherein the shake-bed expansion culture of the strain in the first step comprises:
inoculating the flat seeds on an eggplant bottle slant NA culture medium, and culturing at 28 ℃ for 36h to obtain eggplant bottle seeds.
7. The use of Paenibacillus polymyxa strain P21 in the preparation of an aqueous solution for controlling clubroot of cruciferous vegetables and root rot of Panax notoginseng according to claim 3, wherein the preparation of the seed suspension in step two comprises:
the colonies on the slant surface of the eggplant bottle were washed with sterilized water to obtain a seed suspension.
8. The use of paenibacillus polymyxa strain P21 in the preparation of an aqueous solution for controlling clubroot of cruciferous vegetables and root rot of panax notoginseng according to claim 3, wherein the inoculation in step two comprises:
the seed suspension is inoculated into a fermentation tank culture medium by differential pressure through an inoculation port;
the formula and the processing method of the fermentation tank culture medium are as follows: peptone 0.5%, glucose 0.7%, yeast extract 0.1%, magnesium sulfate 0.15%, dipotassium hydrogen phosphate 0.15%, glycerol 10%, distilled water 1000mL, pH 7.5 before sterilization.
9. The use of paenibacillus polymyxa strain P21 in the preparation of an aqueous solution for preventing and treating clubroot of cruciferous vegetables and root rot of panax notoginseng according to claim 3, wherein the liquid fermentation in the second step comprises:
after inoculation, culturing the fermentation tank culture medium for 36-48 h; wherein the hairThe temperature of the fermentation tank is 26-32 ℃, and the pressure of the fermentation tank is 0.5kg/cm 2 The stirring speed in the tank is 180 rpm; in the process of inoculation and pressure increase, the tank pressure does not exceed 1.5kg/cm 2 。
10. The application of the paenibacillus polymyxa strain P21 in preparing an aqueous solution for preventing and treating clubroot disease of cruciferous vegetables and root rot of panax notoginseng according to claim 2, wherein the paenibacillus polymyxa strain P21 aqueous solution is applied by root irrigation.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202210520768.6A CN114958660B (en) | 2022-05-12 | 2022-05-12 | Paenibacillus polymyxa strain and application thereof |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202210520768.6A CN114958660B (en) | 2022-05-12 | 2022-05-12 | Paenibacillus polymyxa strain and application thereof |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN114958660A true CN114958660A (en) | 2022-08-30 |
| CN114958660B CN114958660B (en) | 2023-12-15 |
Family
ID=82983293
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202210520768.6A Active CN114958660B (en) | 2022-05-12 | 2022-05-12 | Paenibacillus polymyxa strain and application thereof |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN114958660B (en) |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN117904010A (en) * | 2024-03-20 | 2024-04-19 | 中国农业科学院植物保护研究所 | Paenibacillus polymyxa IPP209-1 and application thereof |
Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103667134A (en) * | 2013-12-10 | 2014-03-26 | 江苏省农业科学院 | Paenibacillus polymyxa N3-4 used for preventing and treating clubroot of cruciferous crops and application thereof |
| WO2016019480A1 (en) * | 2014-08-08 | 2016-02-11 | Universidad De Santiago De Chile | Paenibacillus polymyxa schc 33 bacterial strain, and use thereof to combat phytopathogenic fungi in fruits, vegetables or plants |
-
2022
- 2022-05-12 CN CN202210520768.6A patent/CN114958660B/en active Active
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103667134A (en) * | 2013-12-10 | 2014-03-26 | 江苏省农业科学院 | Paenibacillus polymyxa N3-4 used for preventing and treating clubroot of cruciferous crops and application thereof |
| WO2016019480A1 (en) * | 2014-08-08 | 2016-02-11 | Universidad De Santiago De Chile | Paenibacillus polymyxa schc 33 bacterial strain, and use thereof to combat phytopathogenic fungi in fruits, vegetables or plants |
Non-Patent Citations (1)
| Title |
|---|
| 陈雪丽;王光华;金剑;吕宝林;: "多粘类芽孢杆菌BRF-1和枯草芽孢杆菌BRF-2对黄瓜和番茄枯萎病的防治效果" * |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN117904010A (en) * | 2024-03-20 | 2024-04-19 | 中国农业科学院植物保护研究所 | Paenibacillus polymyxa IPP209-1 and application thereof |
| CN117904010B (en) * | 2024-03-20 | 2024-05-31 | 中国农业科学院植物保护研究所 | Paenibacillus polymyxa IPP209-1 and application thereof |
Also Published As
| Publication number | Publication date |
|---|---|
| CN114958660B (en) | 2023-12-15 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN107446847B (en) | Bacillus belgii GT11 and application thereof | |
| CN105296381B (en) | One bacillus subtilis CYY-25 and its application | |
| CN112266881B (en) | Bacillus amyloliquefaciens and application thereof in preventing and treating apple continuous cropping obstacle | |
| CN106754557B (en) | Bacillus subtilis YBM-4 and application thereof in preventing and treating tobacco black shank and promoting growth | |
| CN105838656B (en) | The degeneration-resistant bacillus of broad-spectrum disease resistance growth-promoting and its application for preventing and treating graw mold of tomato and leaf mold | |
| CN101760438B (en) | Broad-spectrum antifungal plant endophytic bacillus subtillis and application thereof | |
| CN107779420B (en) | Two-strain endogenous Bacillus belgii for antagonizing tobacco bacterial wilt and application thereof | |
| CN112358974B (en) | Plant endophytic fungus epicoccum nigrum FZT214 and application thereof | |
| CN103642734B (en) | Microbacterium maritypicum and application thereof in preventing sugar beet disease-causing organisms | |
| CN105543132A (en) | Bacillus methylotrophicus YB-F7 and application thereof in preventing plant diseases | |
| CN111073825B (en) | Bacterium with plant soil-borne disease resistance effect and application thereof | |
| CN113801808B (en) | Streptomyces albocongensis and application thereof | |
| CN102168042A (en) | Pseudomonas aeruginosa and application thereof | |
| CN109112069B (en) | Biocontrol endophytic fungus and application thereof | |
| CN103013852A (en) | Actinomycete capable of inhibiting phytophthoramelonis and screening method thereof | |
| CN109303067B (en) | Streptomyces composition for preventing and treating potato scab and application thereof | |
| CN108913625B (en) | Salt-tolerant streptomycete, microbial inoculum thereof and application of microbial inoculum thereof in promoting plant growth | |
| CN110317747A (en) | A kind of bacillus amyloliquefaciens JT68 and its application in prevention and treatment tea anthracnose | |
| CN107467075B (en) | Application of bacillus pumilus as rice growth promoter | |
| CN114958660A (en) | Paenibacillus polymyxa strain and application thereof | |
| CN111334458B (en) | Biocontrol actinomycetes and application thereof in prevention and control of ginger stem basal rot or soybean epidemic disease | |
| CN106635926B (en) | Bacillus safensis YJC-4 and application thereof in prevention and treatment of tobacco black shank and growth promotion | |
| CN116121147B (en) | Chenopodium ambrosioides seed endophytic Larimol agrobacterium and application thereof | |
| CN114456973B (en) | Streptomyces rochei in tobacco and application thereof in prevention and control of tobacco diseases | |
| CN110205258B (en) | Streptomyces bacterial strain PBS9 for preventing and treating potato scab 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 |