CN116694505A - Streptomyces crimson capable of preventing and treating clubroot and application thereof - Google Patents
Streptomyces crimson capable of preventing and treating clubroot and application thereof Download PDFInfo
- Publication number
- CN116694505A CN116694505A CN202310392822.8A CN202310392822A CN116694505A CN 116694505 A CN116694505 A CN 116694505A CN 202310392822 A CN202310392822 A CN 202310392822A CN 116694505 A CN116694505 A CN 116694505A
- Authority
- CN
- China
- Prior art keywords
- streptomyces
- crimson
- clubroot
- chinese cabbage
- root
- 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
- 241000187747 Streptomyces Species 0.000 title claims abstract description 43
- 238000000855 fermentation Methods 0.000 claims abstract description 27
- 230000004151 fermentation Effects 0.000 claims abstract description 27
- 238000000034 method Methods 0.000 claims abstract description 11
- 239000002689 soil Substances 0.000 claims abstract description 9
- 238000009629 microbiological culture Methods 0.000 claims abstract description 5
- 238000011282 treatment Methods 0.000 claims description 13
- 238000012258 culturing Methods 0.000 claims description 6
- 239000001963 growth medium Substances 0.000 claims description 6
- 230000002262 irrigation Effects 0.000 claims description 6
- 238000003973 irrigation Methods 0.000 claims description 6
- 238000009630 liquid culture Methods 0.000 claims description 5
- 239000002609 medium Substances 0.000 claims description 3
- 230000000737 periodic effect Effects 0.000 claims description 2
- 244000038559 crop plants Species 0.000 claims 1
- 230000000694 effects Effects 0.000 abstract description 19
- 235000010149 Brassica rapa subsp chinensis Nutrition 0.000 abstract description 16
- 235000000536 Brassica rapa subsp pekinensis Nutrition 0.000 abstract description 16
- 241000499436 Brassica rapa subsp. pekinensis Species 0.000 abstract description 16
- 230000005764 inhibitory process Effects 0.000 abstract description 14
- 201000010099 disease Diseases 0.000 abstract description 11
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 abstract description 11
- 230000035784 germination Effects 0.000 abstract description 6
- 241001503464 Plasmodiophora Species 0.000 abstract description 5
- 238000011161 development Methods 0.000 abstract description 5
- 230000018109 developmental process Effects 0.000 abstract description 5
- 210000004209 hair Anatomy 0.000 abstract description 5
- 208000015181 infectious disease Diseases 0.000 abstract description 5
- 230000002265 prevention Effects 0.000 abstract description 5
- 239000002207 metabolite Substances 0.000 abstract description 4
- 239000000575 pesticide Substances 0.000 abstract description 4
- 244000000005 bacterial plant pathogen Species 0.000 abstract description 3
- 239000000126 substance Substances 0.000 abstract description 3
- 231100000331 toxic Toxicity 0.000 abstract description 2
- 230000002588 toxic effect Effects 0.000 abstract description 2
- 241000186361 Actinobacteria <class> Species 0.000 description 8
- 241000196324 Embryophyta Species 0.000 description 8
- 241000894006 Bacteria Species 0.000 description 7
- 230000003042 antagnostic effect Effects 0.000 description 7
- 244000052616 bacterial pathogen Species 0.000 description 6
- 244000053095 fungal pathogen Species 0.000 description 6
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 6
- 241000233866 Fungi Species 0.000 description 4
- 230000001580 bacterial effect Effects 0.000 description 4
- 230000012010 growth Effects 0.000 description 4
- 238000004321 preservation Methods 0.000 description 4
- 239000000725 suspension Substances 0.000 description 4
- QTBSBXVTEAMEQO-UHFFFAOYSA-N Acetic acid Chemical compound CC(O)=O QTBSBXVTEAMEQO-UHFFFAOYSA-N 0.000 description 3
- 102000004157 Hydrolases Human genes 0.000 description 3
- 108090000604 Hydrolases Proteins 0.000 description 3
- 230000000844 anti-bacterial effect Effects 0.000 description 3
- 230000001276 controlling effect Effects 0.000 description 3
- 238000011081 inoculation Methods 0.000 description 3
- 235000015097 nutrients Nutrition 0.000 description 3
- 238000004382 potting Methods 0.000 description 3
- 239000000047 product Substances 0.000 description 3
- 238000011160 research Methods 0.000 description 3
- 238000012360 testing method Methods 0.000 description 3
- 241001446247 uncultured actinomycete Species 0.000 description 3
- 108020004465 16S ribosomal RNA Proteins 0.000 description 2
- UHPMCKVQTMMPCG-UHFFFAOYSA-N 5,8-dihydroxy-2-methoxy-6-methyl-7-(2-oxopropyl)naphthalene-1,4-dione Chemical compound CC1=C(CC(C)=O)C(O)=C2C(=O)C(OC)=CC(=O)C2=C1O UHPMCKVQTMMPCG-UHFFFAOYSA-N 0.000 description 2
- 229920000742 Cotton Polymers 0.000 description 2
- 206010059866 Drug resistance Diseases 0.000 description 2
- 241000223218 Fusarium Species 0.000 description 2
- 206010028980 Neoplasm Diseases 0.000 description 2
- 241000224016 Plasmodium Species 0.000 description 2
- 230000015572 biosynthetic process Effects 0.000 description 2
- 239000011521 glass Substances 0.000 description 2
- 238000004519 manufacturing process Methods 0.000 description 2
- 244000005700 microbiome Species 0.000 description 2
- 230000000877 morphologic effect Effects 0.000 description 2
- 230000000284 resting effect Effects 0.000 description 2
- 238000012216 screening Methods 0.000 description 2
- 229930000044 secondary metabolite Natural products 0.000 description 2
- 230000028327 secretion Effects 0.000 description 2
- 230000004763 spore germination Effects 0.000 description 2
- FWMNVWWHGCHHJJ-SKKKGAJSSA-N 4-amino-1-[(2r)-6-amino-2-[[(2r)-2-[[(2r)-2-[[(2r)-2-amino-3-phenylpropanoyl]amino]-3-phenylpropanoyl]amino]-4-methylpentanoyl]amino]hexanoyl]piperidine-4-carboxylic acid Chemical compound C([C@H](C(=O)N[C@H](CC(C)C)C(=O)N[C@H](CCCCN)C(=O)N1CCC(N)(CC1)C(O)=O)NC(=O)[C@H](N)CC=1C=CC=CC=1)C1=CC=CC=C1 FWMNVWWHGCHHJJ-SKKKGAJSSA-N 0.000 description 1
- 241000186046 Actinomyces Species 0.000 description 1
- 241000187643 Amycolatopsis Species 0.000 description 1
- 235000017166 Bambusa arundinacea Nutrition 0.000 description 1
- 235000017491 Bambusa tulda Nutrition 0.000 description 1
- 241000219198 Brassica Species 0.000 description 1
- 235000003351 Brassica cretica Nutrition 0.000 description 1
- 240000007124 Brassica oleracea Species 0.000 description 1
- 235000003899 Brassica oleracea var acephala Nutrition 0.000 description 1
- 235000011301 Brassica oleracea var capitata Nutrition 0.000 description 1
- 235000001169 Brassica oleracea var oleracea Nutrition 0.000 description 1
- 235000003343 Brassica rupestris Nutrition 0.000 description 1
- 241000219193 Brassicaceae Species 0.000 description 1
- 241000946786 Kitasatospora purpeofusca Species 0.000 description 1
- 238000012408 PCR amplification Methods 0.000 description 1
- 244000082204 Phyllostachys viridis Species 0.000 description 1
- 235000015334 Phyllostachys viridis Nutrition 0.000 description 1
- 244000088415 Raphanus sativus Species 0.000 description 1
- 235000006140 Raphanus sativus var sativus Nutrition 0.000 description 1
- 238000010521 absorption reaction Methods 0.000 description 1
- 238000004458 analytical method Methods 0.000 description 1
- 238000013459 approach Methods 0.000 description 1
- 239000003899 bactericide agent Substances 0.000 description 1
- 239000011425 bamboo Substances 0.000 description 1
- 230000000443 biocontrol Effects 0.000 description 1
- 239000012620 biological material Substances 0.000 description 1
- 230000005540 biological transmission Effects 0.000 description 1
- QKSKPIVNLNLAAV-UHFFFAOYSA-N bis(2-chloroethyl) sulfide Chemical compound ClCCSCCCl QKSKPIVNLNLAAV-UHFFFAOYSA-N 0.000 description 1
- LTINZAODLRIQIX-FBXRGJNPSA-N cefpodoxime proxetil Chemical compound N([C@H]1[C@@H]2N(C1=O)C(=C(CS2)COC)C(=O)OC(C)OC(=O)OC(C)C)C(=O)C(=N/OC)\C1=CSC(N)=N1 LTINZAODLRIQIX-FBXRGJNPSA-N 0.000 description 1
- 230000032823 cell division Effects 0.000 description 1
- 239000013043 chemical agent Substances 0.000 description 1
- 239000003795 chemical substances by application Substances 0.000 description 1
- 238000010276 construction Methods 0.000 description 1
- 239000006059 cover glass Substances 0.000 description 1
- 230000007812 deficiency Effects 0.000 description 1
- 238000001514 detection method Methods 0.000 description 1
- 239000012153 distilled water Substances 0.000 description 1
- 238000004043 dyeing Methods 0.000 description 1
- 238000001962 electrophoresis Methods 0.000 description 1
- 230000007613 environmental effect Effects 0.000 description 1
- 238000001914 filtration Methods 0.000 description 1
- 238000007710 freezing Methods 0.000 description 1
- 230000008014 freezing Effects 0.000 description 1
- 239000005556 hormone Substances 0.000 description 1
- 229940088597 hormone Drugs 0.000 description 1
- 238000005286 illumination Methods 0.000 description 1
- 238000001727 in vivo Methods 0.000 description 1
- 230000006799 invasive growth in response to glucose limitation Effects 0.000 description 1
- 238000005304 joining Methods 0.000 description 1
- 239000007788 liquid Substances 0.000 description 1
- 244000144972 livestock Species 0.000 description 1
- 230000002503 metabolic 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
- 235000010460 mustard Nutrition 0.000 description 1
- 238000011392 neighbor-joining method Methods 0.000 description 1
- 230000003287 optical effect Effects 0.000 description 1
- 238000002360 preparation method Methods 0.000 description 1
- 238000000926 separation method Methods 0.000 description 1
- 238000012163 sequencing technique Methods 0.000 description 1
- 238000010186 staining Methods 0.000 description 1
- 238000003756 stirring Methods 0.000 description 1
- 239000006228 supernatant Substances 0.000 description 1
- 208000024891 symptom Diseases 0.000 description 1
- 238000003786 synthesis reaction Methods 0.000 description 1
- 231100000419 toxicity Toxicity 0.000 description 1
- 230000001988 toxicity Effects 0.000 description 1
- 238000005406 washing Methods 0.000 description 1
- 238000009736 wetting Methods 0.000 description 1
- 238000004383 yellowing Methods 0.000 description 1
Classifications
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
- C12N1/20—Bacteria; Culture media therefor
- C12N1/205—Bacterial isolates
-
- 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
- A01N63/28—Streptomyces
-
- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01P—BIOCIDAL, PEST REPELLANT, PEST ATTRACTANT OR PLANT GROWTH REGULATORY ACTIVITY OF CHEMICAL COMPOUNDS OR PREPARATIONS
- A01P3/00—Fungicides
-
- 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
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12R—INDEXING SCHEME ASSOCIATED WITH SUBCLASSES C12C - C12Q, RELATING TO MICROORGANISMS
- C12R2001/00—Microorganisms ; Processes using microorganisms
- C12R2001/01—Bacteria or Actinomycetales ; using bacteria or Actinomycetales
- C12R2001/465—Streptomyces
Landscapes
- Life Sciences & Earth Sciences (AREA)
- Engineering & Computer Science (AREA)
- Health & Medical Sciences (AREA)
- Zoology (AREA)
- Chemical & Material Sciences (AREA)
- Wood Science & Technology (AREA)
- Biotechnology (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Genetics & Genomics (AREA)
- Organic Chemistry (AREA)
- General Health & Medical Sciences (AREA)
- Microbiology (AREA)
- Virology (AREA)
- Pest Control & Pesticides (AREA)
- Biomedical Technology (AREA)
- Environmental Sciences (AREA)
- Plant Pathology (AREA)
- General Engineering & Computer Science (AREA)
- Medicinal Chemistry (AREA)
- Tropical Medicine & Parasitology (AREA)
- Biochemistry (AREA)
- Dentistry (AREA)
- Chemical Kinetics & Catalysis (AREA)
- General Chemical & Material Sciences (AREA)
- Agronomy & Crop Science (AREA)
- Mycology (AREA)
- Agricultural Chemicals And Associated Chemicals (AREA)
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
Abstract
The invention relates to Streptomyces crimson CC2-6 capable of preventing and treating clubroot, which is preserved in China general microbiological culture Collection center (CGMCC) No.26458 in 2023 and 1-12 days, and also relates to application of the Streptomyces crimson CC2-6 in clubroot. The Streptomyces roseoformis CC2-6 has strong inhibition effect on various plant pathogenic bacteria on a flat plate, and the fermentation liquor metabolite has good prevention and control effects on clubroot. The germination inhibition rate of the dormant spores of the plasmodiophora radicis is 51.01%, the root and root hair infection inhibition rate of the Chinese cabbage is 77.41%, the pot control effect of the Chinese cabbage treated by the CC2-6 fermentation liquor is 27.46%, and the pot control effect of the Chinese cabbage is 53.20% when the Chinese cabbage is treated in 7d in advance. In addition, the strain has natural harmony and compatibility with natural ecology, and has no toxic or side effect and no residue on soil ecology compared with chemical pesticides. Therefore, the method has potential commercial development and application value in biological control practice of diseases.
Description
Technical Field
The invention relates to the field of crop disease control, in particular to Streptomyces roseoformus capable of controlling clubroot and application thereof.
Background
Clubroot is a soil-borne disease that mainly infects some commercial crops in cruciferae, such as rape, radish, mustard tuber, cabbage, and the like. Clubroot is widely distributed in more than fifty countries and regions of the world, including africa, south america, north america, europe, asia and oceangoin, and crop yield losses of up to 10-15% each year around the world due to clubroot, and therefore clubroot is regarded as one of the important cruciferous crop diseases. According to statistics, the occurrence area of the clubroot in China can reach 4800-6000 ten thousand mu in the lighter year, and the hazard area is higher than 1.35 hundred million mu in the outbreak year.
Clubroot disease occurs and produces a hazard at the root of the plant and can be infested by the clubroot bacteria throughout the growth period of the plant, but the seedling stage is the most predominant disease-sensing stage. After the underground root is infected by the plasmodiophora, the hormone change in the plant body is caused, the plant cell division is accelerated, and the main root and the lateral root expand to form tumors. The root surface is smooth in the initial stage of the formation of the swollen root, the root rot in the later stage is changed into dark brown in color, odor is emitted, tumors are enlarged, root hairs are reduced or even absent, and the absorption and the transmission of moisture and nutrients of plants are inhibited, so that the overground parts can show symptoms such as water deficiency wilting leaves yellowing and the like, and the whole plant is withered when serious.
At present, the prevention and treatment of the clubroot still takes chemical agents as main agents, the operation is simple and quick, the cost is low, the effect is quick, but the use of chemical bactericides is easy to cause serious consequences such as drug resistance of pathogenic bacteria, pollution to the environment, residual toxicity and the like, and finally the problems of poor prevention and treatment effect, ecological safety hazard, human health and the like occur.
With further development of scientific research, the biological pesticide prepared by using the secondary metabolite of the biological pesticide has the characteristics of no pollution, no residue, difficult generation of drug resistance of harmful organisms, high environmental compatibility, safety to people and livestock and the like; and the metabolic products generated by microorganisms are utilized to treat plant pathogenic bacteria, so that the research of biological disease control is a current hot spot. The biocontrol potential of secondary metabolites produced by actinomycetes mainly shows the inhibition effect on the growth of hyphae, spore germination and infection of pathogenic bacteria, more and more actinomycetes which are derived from various plant endophytes or different extreme environments are discovered, and the types of the metabolites produced by actinomycetes are also becoming more and more abundant. Therefore, the development of biological control research is an important approach for sustainable development of agriculture, and has important practical significance in production.
Disclosure of Invention
In order to solve the problems, the invention provides the Streptomyces roseoformus which can prevent and treat clubroot, and the Streptomyces roseoformus is preserved in China general microbiological culture collection center (CGMCC) No.26458 in the year 2023, month 1 and day 12.
The invention also provides application of the streptomyces crimson-brown fungus in preventing and treating clubroot.
The invention also provides a method for preventing and treating clubroot, which comprises the step of applying the Streptomyces roseoformis culture product to soil for crop planting.
In a specific embodiment, the method comprises the steps of:
s1: culturing the streptomyces crimson and preparing a streptomyces crimson fermentation broth;
s2: applying the Streptomyces roseoformus fermentation broth to the soil in which the crop is planted.
In a specific embodiment, in S1, the streptomyces roseoformis is cultured in medium No. 1.
In a specific embodiment, S1 comprises the steps of:
s11: inoculating the Streptomyces crimson to a liquid culture medium of Gao's No. 1 for culture;
s12: culturing at 28deg.C and 180rpm for 7 days to obtain the Streptomyces roseoformis fermentation broth.
In a specific embodiment, in S2, the crop is subjected to a periodic rooting treatment by the fermentation broth of streptomyces roseoformis.
In a specific embodiment, the time interval between every two root irrigation treatments is 7 days.
The Streptomyces roseoformis CC2-6 strain has strong inhibition effect on various plant pathogenic bacteria on a flat plate, and fermentation liquor metabolite has good prevention and control effects on clubroot. The germination inhibition rate of the dormant spores of the rhizomatous bacteria is 51.01%, the water-cultured seedling test shows that after the treatment of the fermentation liquor of CC2-6 for 9d, the root hair infection inhibition rate of the Chinese cabbage root is 77.41%, the living body inoculation test shows that after the fermentation liquor of the rhizomatous bacteria and the actinomyces are inoculated for 50d simultaneously, the pot control effect of the Chinese cabbage treated by the fermentation liquor of CC2-6 is 27.46%, and the pot control effect of the Chinese cabbage is 53.20% after the treatment of the pot control effect of the Chinese cabbage is advanced for 7d. In addition, the strain is obtained by separating from soil, has natural harmony and compatibility with natural ecology, and has no toxic or side effect and no residue on the soil ecology compared with chemical pesticides. Therefore, the method has potential commercial development and application value in biological control practice of diseases.
Preservation of biological materials
The purified Streptomyces crimson and brown fungus CC2-6 is preserved in China general microbiological culture Collection center (CGMCC) of China national institute of microbiological culture Collection center (CGMCC) of national academy of sciences of China, no. 3, national area of the Korean, beijing, 1 month and 12 days in 2023, with a preservation number of CGMCC No.26458.
Drawings
FIG. 1 is a photograph of a plate morphology of Streptomyces roseoformus CC 2-6;
FIG. 2 is a photograph of a single colony morphology of Streptomyces roseoformis CC 2-6;
FIG. 3 is a photomicrograph of spore chains and aerial hyphae of Streptomyces roseoformis CC 2-6;
FIG. 4 shows the identification of Streptomyces crimson CC2-6 for the production of various hydrolases;
FIG. 5 shows the inhibition of Streptomyces crimson CC2-6 against pathogenic bacteria of cotton fusarium wilt;
fig. 6 shows the disease controlling effect of Streptomyces roseoformus CC2-6 clubroot.
Detailed Description
The principles and features of the present invention are described below with reference to the drawings, the examples are illustrated for the purpose of illustrating the invention and are not to be construed as limiting the scope of the invention.
1. Identification and preservation of Streptomyces crimson CC2-6
The group collects rhizosphere soil of non-farmland ecological systems such as lawns, bamboo forests, street trees and the like from 4 areas of north, sand-ground dams, banan and south banks of Chongqing, a plurality of strains obtained by separation are subjected to primary screening and secondary screening, one strain CC2-6 is separated, and the strain is determined to have better prevention and treatment effects on clubroot through in-vivo plant indoor potting inoculation tests.
Strains CC2-6 were cultured on a culture plate of Gao's No. 1 at 28℃for 5-7d, and the morphology of the plate was shown in FIG. 1, and the single colony was shown in FIG. 2.
The slide glass sterilized in advance is obliquely inserted into a flat plate at 30 ℃, after being inversely cultured for 7d in a 28 ℃ incubator, the cover glass is taken out, a little distilled water is dripped on the slide glass, and the morphology of the morphological characteristics of aerial hyphae and spore chains of the strain is observed by using an optical microscope, as shown in figure 3.
Genomic DNA of the strain was extracted with bacterial universal primers 27F (5'-AGAGTTTGATCCTGGCTCAG-3', SEQ ID NO: 1) and 1492R (5'-TACGGCTACC TTGACGAC-3', SEQ ID NO: 2). The genome DNA of the strain CC2-6 is used as a template, and after PCR amplification, the 16S rDNA sequence is obtained through electrophoresis detection and sequencing, and is shown as SEQ ID NO. 3. Blast comparison is carried out on the obtained sequence result in NCBI, a 16S rDNA sequence of a related strain is selected, a Neighbor-Joining method (Neighbor-Joining) of MEGA 7.0 software is used for tree construction, and the strain is identified to belong to Streptomyces roseoformis by combining morphological characteristics (Streptomyces purpeofuscus).
The strain is preserved in China general microbiological culture Collection center (CGMCC) of China national institute of culture Collection of microorganisms, national academy of sciences of China No. 3, including North Chen and West Lu No. 1, the Korean region of Beijing, at 1 month 12 of 2023, with a preservation number: CGMCC No.26458.
2. Streptomyces roseofaciens CC2-6 fermentation broth preparation and metabolite analysis
The antagonistic actinomycetes XDS3-6 was inoculated into a sterile liquid medium for Kyoho synthesis No. 1, and subjected to shaking fermentation at 28℃and 180rpm for 7d. The fermentation broth was centrifuged at 7500rpm for 10min, and the supernatant was discarded. Freezing at-80deg.C, and lyophilizing in a lyophilizing machine for 12 hr.
3. Streptomyces crimson CC2-6 for preventing and controlling pathogenic bacteria
3.1 bacteriostasis of Streptomyces crimson CC2-6 against pathogenic bacteria
Antagonistic actinomycetes were inoculated in some hydrolase selection media. The plates were placed upside down in a 28℃incubator and after 7d the strain growth was observed and the radius of the transparent circle was recorded. Streptomyces roseofaciens CC2-6 was detected to produce various hydrolases (FIG. 4). It is presumed that it has a good antibacterial effect against some pathogenic bacteria.
Further, the effect of antagonistic bacteria on the growth of hyphae of pathogenic fungi (fusarium wilt of cotton) was measured by a plate-counter streaking method. The center of the plate is inoculated with a fungus cake of 8mm, antagonistic actinomycetes are inoculated at two sides 25mm away from the fungus cake, two straight lines of 30mm are drawn, actinomycetes are inoculated 3d earlier than the pathogenic fungi, the control group is inoculated with the pathogenic fungi at the center of the plate, actinomycetes are not inoculated, 3 replicates are treated each, and the culture is carried out in a constant temperature incubator at 28 ℃. And when the control group pathogenic fungi grow on the flat plate, observing whether the treatment group generates a zone of inhibition, measuring the colony diameter of the pathogenic fungi, and calculating the inhibition rate. Determining the inhibition effect of the CC2-6 strain on pathogenic fungi. The results are shown in FIG. 5, and the CC2-6 strain has remarkable antibacterial effect.
3.2 Streptomyces crimson CC2-6 for preventing and treating clubroot
Taking root secretion of Chinese cabbage as a culture medium, adding 5mL of root secretion, 0.5mL of suspension of dormant spores of plasmodium and 0.5mL of fermentation liquor of bacterial strain CC2-6 into a sterilized triangular flask, culturing in the dark at 24 ℃, and taking sterilized liquid culture medium as a control, wherein each treatment is repeated for 3 groups. And 5d, observing germination of dormant spores by a microscope, staining with 1% lichen red (dissolved in 45% acetic acid) for 10-15s, and calculating spore germination rate and germination inhibition rate. The results showed that the germination inhibition rate of the dormant spores of the plasmodium rhizomatosis was 51.01%.
Placing Chinese cabbage seeds in a culture dish filled with wet filter paper, accelerating germination under 24 deg.C illumination and wetting condition, transplanting seedlings into 10mL centrifuge tube filled with Hoagland nutrient solution for water culture after 3d, inoculating rhizomatous resting spores and CC2-6 strain fermentation liquor after 5d, wherein the final concentration of rhizomatous resting spores is 10 7 The treated group was added to the broth at a ratio of 1:30 per mL, and the control group was added to Hoagland nutrient solution. And after water planting for 9d, taking the root of the seedling, washing with clear water, putting the seedling into fluorescent pink dye liquor for dyeing for 30min, and moving under a microscope to observe the infection condition of the root hair of the seedling. The result shows that after the Chinese cabbage is treated by the CC2-6 fermentation liquor for 9 days, the root hair infection inhibition rate of the Chinese cabbage root is 77.41%.
Inoculating Chinese cabbage by root irrigation. Taking out the root from refrigerator, placing at 25deg.C, rotting in dark environment for 5 days, mixing the root with water at a ratio of 1:5, intermittently stirring with juicer, filtering, and preparing and adjusting the concentration of the dormant spore suspension of the root of the Amycolatopsis to 2.5X10 8 And each mL. Inoculating antagonistic bacterial cake into Gao's first liquid culture medium at 28deg.C and 180r.min -1 Culturing under the condition for 7d to obtain the product with concentration of 10 7 CFU/mL actinomycete fermentation broth. Root irrigation treatment of each Chinese cabbage with 30mL of dormant spore suspension of plasmodiophora radicis to obtain final bacterial content of 3×10 per gram of soil 7 And each. Treatment a: antagonistic bacteria and plasmodiophora radicis are inoculated simultaneously, 20mL of actinomycete fermentation liquor is irrigated every 7d, and the total irrigation is carried out five times, and the treatment B: the antagonistic bacteria are inoculated in advance for 7 days, 20mL of actinomycete fermentation liquor is irrigated every 7 days, and the total irrigation is five times. Inoculating the suspension of dormant spores of the rhizomatous bacteria and irrigating a liquid culture medium of Gao's first as a disease control, and repeating 20 white strains each treatmentThe disease was investigated 50d after inoculation and statistically recorded. The results show that after the plasmodiophora radicis and the CC2-6 fermentation liquor are inoculated for 50 days simultaneously, the control effect of the Chinese cabbage potting treated by the CC2-6 fermentation liquor is 27.46%, and the control effect of the Chinese cabbage potting treated by the CC2-6 fermentation liquor is 53.20% when the CC2-6 fermentation liquor is treated for 7 days in advance (figure 6).
The foregoing description of the preferred embodiments of the invention is not intended to limit the invention to the precise form disclosed, and any such modifications, equivalents, and alternatives falling within the spirit and scope of the invention are intended to be included within the scope of the invention.
Claims (8)
1. Streptomyces crimson for preventing and treating clubroot is characterized in that the Streptomyces crimson is preserved in China general microbiological culture Collection center (CGMCC No. 26458) in 1-12-year 2023.
2. The use of streptomyces crimson according to claim 1 for controlling clubroot.
3. A method for controlling clubroot, comprising the step of applying the streptomyces crimson culture product of claim 1 to the soil of crop plants.
4. A method according to claim 3, comprising the steps of:
s1: culturing the streptomyces crimson and preparing a streptomyces crimson fermentation broth;
s2: applying the Streptomyces roseoformus fermentation broth to the soil in which the crop is planted.
5. The method of claim 4, wherein in S1, the streptomyces crimson-brown mold is cultured in medium No. one of gao.
6. The method of claim 5, wherein S1 comprises the steps of:
s11: inoculating the Streptomyces crimson to a liquid culture medium of Gao's No. 1 for culture;
s12: culturing at 28deg.C and 180rpm for 7 days to obtain the Streptomyces roseoformis fermentation broth.
7. The method of claim 4, wherein in S2 the crop is subjected to periodic rooting with the fermentation broth of streptomyces roseoformis.
8. The method of claim 7, wherein the time interval between each root irrigation treatment is 7 days.
Priority Applications (1)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
CN202310392822.8A CN116694505B (en) | 2023-04-13 | 2023-04-13 | Streptomyces crimson capable of preventing and treating clubroot and application thereof |
Applications Claiming Priority (1)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
CN202310392822.8A CN116694505B (en) | 2023-04-13 | 2023-04-13 | Streptomyces crimson capable of preventing and treating clubroot and application thereof |
Publications (2)
Publication Number | Publication Date |
---|---|
CN116694505A true CN116694505A (en) | 2023-09-05 |
CN116694505B CN116694505B (en) | 2024-07-16 |
Family
ID=87844119
Family Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
CN202310392822.8A Active CN116694505B (en) | 2023-04-13 | 2023-04-13 | Streptomyces crimson capable of preventing and treating clubroot and application thereof |
Country Status (1)
Country | Link |
---|---|
CN (1) | CN116694505B (en) |
Cited By (1)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN116064288A (en) * | 2022-08-30 | 2023-05-05 | 内蒙古农业大学 | Streptomyces roseoformis HC7-22 for plant iron-removing and growth-promoting and application thereof |
Citations (9)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US4261979A (en) * | 1978-10-19 | 1981-04-14 | Agence Nationale De Valorisation De La Recherche (Anvar) | Novel water-soluble compounds derived from extracts of streptomyces stimulosus, process for their production, their uses as immunological adjuvants and compositions containing them |
KR100760526B1 (en) * | 2006-04-06 | 2007-10-04 | 한국화학연구원 | 119 119 fusarium oxysporum ef119 strain producing bikaverin microorganism formulation for controlling plant diseases containing bikaverin or ef119 strain and method for controlling plant diseases using same |
KR100942228B1 (en) * | 2009-11-10 | 2010-02-16 | 충청남도 | Biological control of plant diseases using flavobacterium hercynium epb-c313 |
CN102864110A (en) * | 2012-09-27 | 2013-01-09 | 四川农业大学 | Streptomyces purpeofuscus SP3 strain and application thereof in prevention of soil-borne root diseases of Taxus chinensis and growth promotion of Taxus chinensis |
CN103013852A (en) * | 2012-10-10 | 2013-04-03 | 湖南农业大学 | Actinomycete capable of inhibiting phytophthoramelonis and screening method thereof |
CN103088094A (en) * | 2013-01-29 | 2013-05-08 | 四川农业大学 | Streptomyces purpeofuscus SP3 antifungal protain mother liquid and application thereof in preparaiton of induced hybrid bambood disease-resistant insecticide |
US20150191756A1 (en) * | 2012-09-10 | 2015-07-09 | Mitsubishi Rayon Co., Ltd | Method for producing methacrylic acid and/or ester thereof |
CN108812693A (en) * | 2018-05-24 | 2018-11-16 | 林荣铨 | Application of the graphene oxide in terms of improving plant phytophthora germ Antagonistic Fungi preventive effect |
CN115851553A (en) * | 2022-12-30 | 2023-03-28 | 西南大学 | Streptomyces virginiae capable of preventing and treating clubroot and application thereof |
-
2023
- 2023-04-13 CN CN202310392822.8A patent/CN116694505B/en active Active
Patent Citations (9)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US4261979A (en) * | 1978-10-19 | 1981-04-14 | Agence Nationale De Valorisation De La Recherche (Anvar) | Novel water-soluble compounds derived from extracts of streptomyces stimulosus, process for their production, their uses as immunological adjuvants and compositions containing them |
KR100760526B1 (en) * | 2006-04-06 | 2007-10-04 | 한국화학연구원 | 119 119 fusarium oxysporum ef119 strain producing bikaverin microorganism formulation for controlling plant diseases containing bikaverin or ef119 strain and method for controlling plant diseases using same |
KR100942228B1 (en) * | 2009-11-10 | 2010-02-16 | 충청남도 | Biological control of plant diseases using flavobacterium hercynium epb-c313 |
US20150191756A1 (en) * | 2012-09-10 | 2015-07-09 | Mitsubishi Rayon Co., Ltd | Method for producing methacrylic acid and/or ester thereof |
CN102864110A (en) * | 2012-09-27 | 2013-01-09 | 四川农业大学 | Streptomyces purpeofuscus SP3 strain and application thereof in prevention of soil-borne root diseases of Taxus chinensis and growth promotion of Taxus chinensis |
CN103013852A (en) * | 2012-10-10 | 2013-04-03 | 湖南农业大学 | Actinomycete capable of inhibiting phytophthoramelonis and screening method thereof |
CN103088094A (en) * | 2013-01-29 | 2013-05-08 | 四川农业大学 | Streptomyces purpeofuscus SP3 antifungal protain mother liquid and application thereof in preparaiton of induced hybrid bambood disease-resistant insecticide |
CN108812693A (en) * | 2018-05-24 | 2018-11-16 | 林荣铨 | Application of the graphene oxide in terms of improving plant phytophthora germ Antagonistic Fungi preventive effect |
CN115851553A (en) * | 2022-12-30 | 2023-03-28 | 西南大学 | Streptomyces virginiae capable of preventing and treating clubroot and application thereof |
Non-Patent Citations (2)
Title |
---|
SANJAY ANTONY-BADU等: "Multiple streptomyces species with distinct secondary metabolomes have identical 16S rRNA gene sequences", SCI REP, 11 September 2017 (2017-09-11) * |
谯天敏等: "绛红褐链霉菌的根际定殖能力动态监测", 南京林业大学学报(自然科学版), 15 September 2015 (2015-09-15) * |
Cited By (2)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN116064288A (en) * | 2022-08-30 | 2023-05-05 | 内蒙古农业大学 | Streptomyces roseoformis HC7-22 for plant iron-removing and growth-promoting and application thereof |
CN116064288B (en) * | 2022-08-30 | 2024-04-09 | 内蒙古农业大学 | Streptomyces roseoformis HC7-22 for plant iron-removing and growth-promoting and application thereof |
Also Published As
Publication number | Publication date |
---|---|
CN116694505B (en) | 2024-07-16 |
Similar Documents
Publication | Publication Date | Title |
---|---|---|
CN107446847B (en) | Bacillus belgii GT11 and application thereof | |
CN104560827B (en) | A kind of biocontrol actinomycetes bacterial strain for preventing and treating tobacco bacterial wilt and its application | |
CN105543132A (en) | Bacillus methylotrophicus YB-F7 and application thereof in preventing plant diseases | |
CN115058358B (en) | Salt-tolerant bacillus and application thereof | |
CN110200016B (en) | Streptomyces strain PBSH9 capable of preventing and treating potato scab and promoting potato growth and application thereof | |
CN112342173B (en) | Bacillus belgii and application thereof | |
CN110218685A (en) | Biocontrol actinomycetes and application | |
CN116694505B (en) | Streptomyces crimson capable of preventing and treating clubroot and application thereof | |
CN114134068A (en) | Paenibacillus polymyxa microbial agent, preparation method and application thereof | |
CN105567600A (en) | Pathogen verticillium antagonistic bacterium and application thereof | |
CN109628341B (en) | Streptomyces violaceorubidus and biological control microbial inoculum and preparation method thereof | |
CN103146609B (en) | Pseudomonas fluorescens and method for preventing phytophthora capsici thereby | |
CN115851553B (en) | Streptomyces virginiae capable of preventing and treating clubroot and application thereof | |
CN108913621B (en) | Bacillus methylotrophicus YH-18 capable of effectively preventing and treating oriental cherry root cancer and application thereof | |
CN110317747A (en) | A kind of bacillus amyloliquefaciens JT68 and its application in prevention and treatment tea anthracnose | |
CN111334458B (en) | Biocontrol actinomycetes and application thereof in prevention and control of ginger stem basal rot or soybean epidemic disease | |
CN114456973B (en) | Streptomyces rochei in tobacco and application thereof in prevention and control of tobacco diseases | |
CN114933980B (en) | Streptomyces shallowus HJB-XTBG45 for preventing and treating rhizoma polygonati root rot and application thereof | |
CN114703080B (en) | Streptomyces fuscoporia and application thereof | |
CN114032182B (en) | Fungus with functions of antagonizing pathogenic bacteria of garlic root rot and promoting growth | |
CN109055265A (en) | A kind of biocontrol microorganisms and its application in terms of crops blight dis-ease prevention and treatment | |
CN110205258B (en) | Streptomyces bacterial strain PBS9 for preventing and treating potato scab and application thereof | |
CN116144538B (en) | Streptomyces flavochromogenes capable of preventing and treating clubroot and application thereof | |
CN107603904B (en) | Streptomyces and application thereof | |
CN110066755A (en) | One plant of fructus arctii endogenetic streptomycete, the microbial bacterial agent containing the endophyte and application |
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 |