CN105543132A - Bacillus methylotrophicus YB-F7 and application thereof in preventing plant diseases - Google Patents
Bacillus methylotrophicus YB-F7 and application thereof in preventing plant diseases Download PDFInfo
- Publication number
- CN105543132A CN105543132A CN201511026963.XA CN201511026963A CN105543132A CN 105543132 A CN105543132 A CN 105543132A CN 201511026963 A CN201511026963 A CN 201511026963A CN 105543132 A CN105543132 A CN 105543132A
- Authority
- CN
- China
- Prior art keywords
- microbial
- root
- genus bacillus
- disease
- bacterial strain
- 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
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
-
- 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
Landscapes
- Life Sciences & Earth Sciences (AREA)
- Health & Medical Sciences (AREA)
- Engineering & Computer Science (AREA)
- Zoology (AREA)
- Chemical & Material Sciences (AREA)
- Biotechnology (AREA)
- General Health & Medical Sciences (AREA)
- Wood Science & Technology (AREA)
- Organic Chemistry (AREA)
- Virology (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Genetics & Genomics (AREA)
- Microbiology (AREA)
- Tropical Medicine & Parasitology (AREA)
- Biochemistry (AREA)
- General Engineering & Computer Science (AREA)
- Biomedical Technology (AREA)
- Medicinal Chemistry (AREA)
- Agronomy & Crop Science (AREA)
- Pest Control & Pesticides (AREA)
- Plant Pathology (AREA)
- Dentistry (AREA)
- Environmental Sciences (AREA)
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
- Agricultural Chemicals And Associated Chemicals (AREA)
Abstract
The invention discloses bacillus methylotrophicus YB-F7 and an application thereof in preventing plant diseases. The YB-F7 bacterial strain is collected in China General Microbiological Culture Collection Center (CGMCC), and the collection number is CGMCC NO. 10834; and a nucleotide sequence of the bacterial strain is SEQ ID NO.1. The selected and separated bacillus methylotrophicus YB-F7 is a microbial agent having relatively strong suppression on gaeumannomyces graminis, rhizoctonia cerealis and fusarium graminearum soil-borne diseases. The YB-7 agent can be applied during the crop seeding period or the disease beginning period, and a fermenting agent is used for being mixed with seeds or diluted by water to be applied to roots of crops in a root irrigation manner or to be sprayed; and the bacillus methylotrophicus can be mixed with other bactericides and plant regulators having no side effect on the biocontrol strain so as to regulate the microbial community and control the soil-borne diseases of the wheat, thereby realizing the purpose of ecological efficiency.
Description
One, technical field:
The present invention relates to control of plant disease technical field, be specifically related to a kind of Methylotrophic genus bacillus YB-F7 and utilize this genus bacillus controlling plant diseases effect and the application in preparation controlling plant diseases microbial inoculum.
Two, technical background:
At present, microbial control technology is the important component part of modernization Agricultural Sustainable Development, due to the use of chemical pesticide, cause nature long term accumulation objectionable impurities, cause the serious consequence such as environmental pollution, disruption of ecological balance, along with to this useful environment of microbial pesticide, pollution-free, maintain the Study of Prevention Technology of the eubiosis, replace the use of chemical pesticide to become the focus of current microbial control research.
Genus bacillus is a kind of gram-positive microorganism, have growth rapidly, stable, the adaptability of proterties and strong stress resistance, in the application of land for growing field crops, with the obvious advantage compared with other biocontrol microorganisms, environmentally safe nontoxic to people and animals simultaneously; Genus bacillus meta-bolites enriches, and is secreted into outside born of the same parents and can suppresses, kills pathogenic bacteria, or changes its growth of pathogenic bacteria surround inhibition; Some of them bacterial strain can produce multiple antimicrobial substance, and these antimicrobial substances play keying action in biological and ecological methods to prevent plant disease, pests, and erosion process.Genus bacillus is the common biocontrol microorganisms of a class, at present the existing multiple genus bacillus report with the function such as antagonism, growth-promoting.Therefore, screen efficient and activity stabilized antagonistic strain, produce efficient bacteria agent, advance the research of novel microorganism agricultural chemicals, ensure that China's agricultural sustainable development is significant.
Three, summary of the invention:
The technical problem to be solved in the present invention is: provide a kind of Methylotrophic genus bacillus (Bacillusmethylotrophicus) YB-F7 and the application in controlling plant diseases thereof.The application of Methylotrophic genus bacillus (Bacillusmethylotrophicus) YB-F7 of the present invention in controlling plant diseases be specially for biological control gaeumannomyces graminis disease microbial plant soil-borne diseases, application in banded sclerotial blight microbial roots of plants stem's disease or root-rot reaping hook microbial axis basal stem rot evil; Can also for the preparation of the application prevented and treated in gaeumannomyces graminis disease microbial plant soil-borne diseases, banded sclerotial blight microbial roots of plants stem's disease or root-rot reaping hook microbial axis basal stem rot evil microbial inoculum.
The present invention solves the problem by the following technical programs:
The invention provides a kind of Methylotrophic genus bacillus (Bacillusmethylotrophicus) YB-F7, this bacterial strain is preserved in China Committee for Culture Collection of Microorganisms's common micro-organisms center, and deposit number is CGMCCNO.10834.
The preservation information of described Methylotrophic genus bacillus (Bacillusmethylotrophicus) YB-F7 is as follows:
Classification And Nomenclature: Methylotrophic genus bacillus;
Depositary institution: China Microbial Culture Preservation Commission's common micro-organisms center (CGMCC);
Preservation address: No. 3, Yard 1, BeiChen xi Road, Chaoyang District, Beijing City, Institute of Microorganism, Academia Sinica;
Preservation date: on May 21st, 2015;
Deposit number: CGMCCNO.10834.
According to above-mentioned Methylotrophic genus bacillus (Bacillusmethylotrophicus) YB-F7, the nucleotides sequence of described Methylotrophic genus bacillus YB-F7 is classified as SEQIDNO.1.
Methylotrophic genus bacillus (Bacillusmethylotrophicus) YB-F7 of the present invention be used for biological control gaeumannomyces graminis disease microbial plant soil-borne diseases, application in banded sclerotial blight microbial roots of plants stem's disease or root-rot reaping hook microbial axis basal stem rot evil.
Methylotrophic genus bacillus (Bacillusmethylotrophicus) YB-F7 of the present invention is for the preparation of the application prevented and treated in gaeumannomyces graminis disease microbial plant soil-borne diseases, banded sclerotial blight microbial roots of plants stem's disease or root-rot reaping hook microbial axis basal stem rot evil microbial inoculum.
Positive beneficial effect of the present invention:
To be a strain have microbiobacterial agent compared with high inhibition to gaeumannomyces graminis (Gaeumannomycesgraminis), rhizoctonia cerealis (Rhizoctoniacerealis), root-rot sickle-like bacteria (Fusariumgraminearum) soil-borne disease to Methylotrophic genus bacillus (Bacillusmethylotrophicus) YB-F7 that the present invention filters out, and YB-F7 bacterial strain is respectively 90%, 85.5% and 79.8% to gaeumannomyces graminis, rhizoctonia cerealis and root-rot sickle-like bacteria indoor inhibitory effect; Greenhouse pot culture prevents and treats take-all, wheat hypochnus effect reaches 65.7%, 76.5% respectively, and preventive effect is higher than contrast medicament or with to contrast medicament suitable.YB-F7 bacterial strain of the present invention obtains, to Crop securify by being separated in wheat rhizosphere soil; YB-F7 microbial inoculum of the present invention can be used as crop sowing time or the disease phase of starting uses, and adopts fermentation preparation and seed is dressed seed or apply crop rhizosphere or spray application by filling with root after dilute with water; Further, can be used in combination with other sterilant had no side effect to this biocontrol strain, plant modifying agent, regulate soil microorganisms flora, control the object that wheat soil-borne diseases reaches ecological efficient.
Four, accompanying drawing illustrates:
Fig. 1 YB-F7 bacterial strain of the present invention is to the fungistatic effect figure of gaeumannomyces graminis;
Fig. 2 YB-F7 bacterial strain of the present invention is to the fungistatic effect figure of rhizoctonia cerealis;
Fig. 3 YB-F7 bacterial strain of the present invention is to the fungistatic effect figure of Fusarium root rot of wheat;
Fig. 4 biocontrol microorganisms YB-F7 of the present invention prevents and treats wheat hypochnus control efficacy and measures schematic diagram;
Fig. 5 biocontrol microorganisms YB-F7 of the present invention prevents and treats take-all pot experiment schematic diagram;
Fig. 6 YB-F7 bacterial strain of the present invention 16rDNA sequential system grows tree schematic diagram.
Five, embodiment:
Set forth the present invention further below in conjunction with embodiment, but do not limit content of the present invention.
Embodiment 1: the screening of Methylotrophic genus bacillus (Bacillusmethylotrophicus) YB-F7 bacterial strain of the present invention be separated:
For examination pathogenic fungi: gaeumannomyces graminis (Gaeumannomycesgraminis), rhizoctonia cerealis (Rhizoctoniacerealis) and root-rot sickle-like bacteria (Fusariumgraminearum).
Bacteria culture medium:
NA substratum: beef extract 3g, yeast extract paste 1g, peptone 5g, glucose 10g, agar 15g, water 1L, pH6.8 ~ 7.0,121 DEG C of autoclaving 30min.
LB liquid nutrient medium: peptone 10g, yeast extract 5g, NaCl10g, water 1L, pH7.0,121 DEG C of autoclaving 30min.
The isolation and purification of Methylotrophic genus bacillus (Bacillusmethylotrophicus) YB-F7 of the present invention:
The applicant is in planting base, experiment field, Yu Ying village, Henan Academy of Agricultural Sciences's Zhumadian Xiping County, carried out the separation of wheat root disease and rhizospheric microflora of soil diversity change investigation in 2010 ~ 2014 years, in this research trial, antagonistic strain plate screening is separated.Soil collecting is the earthing removing root surface about 5cm from wheat rhizosphere soil, by S shape, the loose earth that random acquisition plant root combines, or from soil, entirety is dug out, employing is trembled local method and is shaked off mixed soil sample, and 3 that are gathered in each place are repeated soil sample and dry in the shade.Take the soil sample that 1g dries in the shade to load in sterilizing test tubes, add 9mL sterilized water, on vortex instrument, abundant vortex 1-2min, makes the soil dilution liquid of 1:10.From gained soil dilution liquid, take out 1mL, add the soil dilution liquid that the abundant vortex of 9mL sterilized water becomes 1:10, the like, make 1:10
3, 1:10
4, 1:10
5soil dilution liquid, respectively get 200 μ L soil dilution liquid in the middle of dull and stereotyped, with spreading rod, spreadable for the bacterium liquid bacterium liquid that makes be distributed in NA planar surface uniformly, be placed in the constant incubator of 28 DEG C, cultivate about 24h.Single bacterium colony of the different colonial morphology of picking, by after microscopy at the flat lining out of NA, 28 DEG C of purifying are cultivated, and it is for subsequent use that the bacterial strain that noticeable change do not occur colony growth characteristic is stored in NA medium slant, namely obtains the Antagonistic Fungi of separation and purification.
Antagonistic Fungi screens:
The Antagonistic Fungi that above-mentioned separation and purification is obtained with pathogenic fungi (i.e. gaeumannomyces graminis, rhizoctonia cerealis and root-rot sickle-like bacteria) for target, adopt dull and stereotyped opposite culture method, the stem of diameter 5mm is inoculated in the dull and stereotyped right-angled intersection centre of PDA, 25 DEG C of incubators are inverted and are cultivated 1-2d, connect 4 kinds of different bacterium, after 25 DEG C of cultivation 2-3d, observe its fungistatic effect, and measure antibacterial band size, select the good bacterial strain of fungistatic effect to carry out multiple sieve, namely obtain the good primary dcreening operation antagonistic strain of fungistatic effect.
Antagonistic strain sieves again:
Prepared by fermented liquid: be seeded to by the antagonistic strain that above-mentioned primary dcreening operation obtains in LB liquid medium, and 24h-48h cultivated by 30 DEG C of shaking tables, gets supernatant liquor for subsequent use.
Bacteria culturing: be this laboratory for examination pathogenic bacteria and preserve, aseptically, be forwarded on PDA flat board from cultured pathogenic bacteria inclined-plane, cultivate 5-7 days, for subsequent use.
By the above-mentioned for subsequent use pathogenic fungi being cultured to full ware, stem is beaten with 5mm punch tool, the dull and stereotyped right-angled intersection centre of access PDA, 25 DEG C of incubators are cultivated and are inverted 1-2d, apart from 3cm place, central surrounding position after taking-up, aseptic technique is vertically put Oxford cup (internal diameter 6mm, high 10mm) in media surface and is gently pressed substratum, it is made to contact tight with substratum, 100uL fermented liquid (i.e. the fermented liquid of above-mentioned preparation) is added in the cup of Oxford, note fermented liquid not being overflowed, after 2-3d cultivated by 25 DEG C of incubators, observe fungistatic effect, measure bacteriostatic diameter.Finally by strain called after Methylotrophic genus bacillus of the present invention YB-F7 bacterial strain the strongest for the fungistatic effect obtained.
The two diameter mean values-bacterium cake diameter of colony diameter (mm)=perpendicular
The qualification of Methylotrophic genus bacillus YB-F7 bacterial strain of the present invention:
To the bacterial strain YB-F7 that the present invention is separated, by Observation of biological characteristics, carry out further identification of morphology, method is as follows:
(1) dye: dye by Gram-staining process;
(2) form and cultural characteristic: cultivate 48h for 28 DEG C on LB solid medium, observe its colony characteristics and color etc., by observation by light microscope thalline feature;
LB solid medium: Tryptones 10g, yeast extract 5g, sodium-chlor 10g, agar 15g, add water to 1000mL;
(3) physiological and biochemical property: carry out with reference to " common bacteria system identification handbook " method;
(4) 16SrDNA and analysis: bacterial strain YB-F7 adopts LB liquid nutrient medium 30 DEG C of shaking culture to logarithmic phase, centrifugal with 10000r/min, collect thalline, conventionally extract bacterial genomes DNA, the general PCR primer of synthesizing for the bacterial 16 S rDNA sequence that increases increases, obtain the amplified production of length 1.4kb, product is carried out reclaiming and checking order, by BLAST, sequencing result is analyzed, carry out tetraploid rice with the sequence in GenBank database, determine the classification position of this bacterial strain.
LB liquid nutrient medium: Tryptones 10g, yeast extract 5g, sodium-chlor 10g, add water to 1000mL;
Above experimental result record is as follows:
1) bacterial strain YB-F7 morphological specificity:
Bacterial strain YB-F7 is circular or ellipse, oyster white, dimpling, neat in edge, individual cells size (0.6 ~ 0.7) × (2 ~ 3) micron on LB substratum.
2) bio-chemical characteristics is the results detailed in Table 1:
Table 1 bacterial strain YB-F7 of the present invention biocontrol microorganisms physicochemical characteristics
| Test subject | Test-results | Test subject | Test-results |
| Gramstaining | + | Urase | - |
| Starch Hydrolysis | + | Lipase | + |
| Gelatin hydrolysis | + | Indoles | + |
| Anaerobic growth | + | Hydrogen sulfide | - |
| Methyl red | - | Glucose | + |
| V-P tests | + | Maltose | + |
| Citrate trianion utilizes | + | Pectinose | + |
| Catalase | + | Wood sugar | + |
| Oxydase | + | Semi-lactosi | + |
| Nitrate reduction | + | N.F,USP MANNITOL | + |
3) Molecular Identification of Methylotrophic genus bacillus YB-F7 of the present invention:
Methylotrophic genus bacillus molecular biology identification of the present invention: with bacterial strain YB-F7 genomic dna for template primer 16srDNA carries out pcr amplification, it is 1443bp that amplification hands over order-checking company to record this strain sequence length, complete sequence submits GenBank to, the search of application BLAST homology, similarity analysis is carried out with strain sequence each in database, in front 100 Search Results, 81 are B.methylotrophicus bacterial strain, and consistence all reaches more than 99%.A class is polymerized to by the sequence of the visible bacterial strain YB-F7 of the phylogenetic tree of the 16SrDNA sequential analysis of bacterial strain YB-F7 and B.methylotrophicus, independent formation branch, genetic distance is the shortest, a tree reliability reaches 100%, in conjunction with physiological and biochemical property, identify that YB-F7 bacterial strain of the present invention is Methylotrophic bacillus methylotrophicus thus.The bacterial strain YB-F7 genome 16rDNA sequence be separated refers to SEQIDNO.1 (YB-F7 bacterial strain 16rDNA sequential system is grown tree and referred to accompanying drawing 6).
Embodiment 3: biocontrol microorganisms YB-F7 of the present invention tests gaeumannomyces graminis, sheath blight fungus and root-rot sickle-like bacteria indoor inhibitory effect:
The present invention adopts face-off, Oxford cup and mycelial growth rate method three kinds of methods in ware, choose representational soil-transmitted disease, rhizome portion disease and brown foot rot disease, determine the fungistatic effect of YB-F7 bacterial strain of the present invention to Pathogen of Take-all, sheath blight fungus and root-rot sickle-like bacteria.Test result is as shown in table 2.Refer to accompanying drawing 1,2,3 in addition, accompanying drawing 1,2,3 is respectively the inhibition display figure of biocontrol microorganisms YB-F7 of the present invention to Pathogen of Take-all, sheath blight fungus and root-rot sickle-like bacteria.
Table 2 YB-F7 bacterial strain of the present invention is to three kinds of pathogenic fungi fungistatic effects
| Pathogenic bacteria | Gaeumannomyces graminis | Rhizoctonia cerealis | Root-rot sickle-like bacteria |
| Antibacterial band (mm) | 13 | 11 | 6 |
| Inhibiting rate (%) | 90.0 | 85.5 | 79.8 |
As can be seen from table 2 and accompanying drawing 1,2,3: YB-F7 bacterial strain of the present invention all has good inhibition to gaeumannomyces graminis, sheath blight fungus and root-rot sickle-like bacteria; Viable bacteria and survey experiment without fermented liquid raw and show, this bacterial strain is obvious to Pathogen of Take-all, sheath blight fungus and root-rot sickle-like bacteria inhibition.So, Methylotrophic genus bacillus (Bacillusmethylotrophicus) YB-F7 of the present invention can be effective to biological control gaeumannomyces graminis disease microbial plant soil-borne diseases, application in banded sclerotial blight microbial roots of plants stem's disease or root-rot reaping hook microbial axis basal stem rot evil.
Embodiment 4: biocontrol microorganisms YB-F7 of the present invention prevents and treats wheat hypochnus control efficacy and measures (see accompanying drawing 4):
(1) experiment material:
1) for examination wheat breed: Zheng wheat 9023;
2) reagent agent: 20% jingganmycin WP;
3) strains tested: rhizoctonia cerealis and biocontrol strain of the present invention be Plant Protection Inst., Henan Agricultural Science Academy's molecular components from.
(2) experimental design:
1) biocontrol microorganisms fermented liquid is cultivated: biocontrol strain of the present invention is seeded to LB liquid nutrient medium (with above-mentioned), 37 DEG C of rotating speed 180r/min, cultivation 48h (what cultivation obtained herein is biocontrol microorganisms fermented liquid stoste, also referred to as antagonistic strain fermented liquid) for subsequent use;
2) rhizoctonia cerealis flat board preparation: by dull and stereotyped for rhizoctonia cerealis good for purifying inoculation PDA, PDA plate culture medium falls 20mL, cultivates about 7d, is paved with flat board to bacterium colony;
3) seed treatment: by wheat seed through 1% hypochlorite disinfectant process 10min after, then use aseptic water washing 3 times, in 25 DEG C of thermostat containers, vernalization is to showing money or valuables one carries unintentionally, and is then soaked in (spore concentration: 10 in the biocontrol microorganisms fermented liquid of the above-mentioned preparation of the present invention
9cfu/ml), soak 3h, be then seeded on the flat board of sheath blight fungus, often process 30, repeat 4 times, after 25 DEG C of cultivation 24h, cover aseptic quartz sand; Establish blank and medicament contrast respectively; Each process is placed in 18-20 DEG C of illumination box, illumination 12h, normally add water every day moisturizing;
4) investigate: after emerging, observe incidence and wheat seedling growth situation, investigation seedling rate, wait the contrast state of an illness relatively stable, the investigation state of an illness, records severity and sickness rate, calculates disease index and prevention effect;
5) wheat hypochnus grade scale: 0 grade: complete stool is anosis; 1 grade: outer leaf sheath less than 1/2 browning; 2 grades: outer leaf sheath more than 1/2 browning; 3 grades: basal part of stem produces obvious eye scab; 4 grades: plant Died Of Disease.
6) preventive effect calculates:
(3) experiment process:
Process 1: blank, sterilized water is soaked seed;
Process 2: medicament contrasts, 20% jingganmycin 1000 times immersion kind;
Process 3: biocontrol microorganisms fermented liquid stoste seed soaking of the present invention;
Process 4: biocontrol microorganisms fermented liquid of the present invention dilutes 5 times of seed soaking;
Process 5: biocontrol microorganisms fermented liquid of the present invention dilutes 10 times of seed soaking.
(4) test-results (referring to table 3):
Table 3 biocontrol microorganisms control of the present invention wheat hypochnus control efficacy measurement result
| Process | Seedling rate (%) | Sickness rate (%) | Disease index (%) | Prevention effect (%) |
| Blank | 85.8 | 90.6 | 52.8 | — |
| 20% jingganmycin | 90.9 | 100 | 40.9 | 22.5 |
| Derosal | 85 | 78.3 | 48.4 | 8.3 |
| YB-F7 stoste | 85 | 64.9 | 30.1 | 43.0 |
| YB-F7 dilution 5 | 84.2 | 76.3 | 18.1 | 65.7 |
| YB-F7 dilution 10 | 91.7 | 70.2 | 35.3 | 33.1 |
As can be seen from table 3 measurement result: biocontrol microorganisms YB-F7 of the present invention has good prevention effect to wheat hypochnus.Control efficacy measurement result shows, fermented liquid dilutes the prevention effect 65.7% of 5 times, higher than 20% jingganmycin and derosal, and stoste and prevention effect difference 43% and 33.1% after diluting 10 times.And bacterium liquid does not affect seedling rate.
Embodiment 5: biocontrol microorganisms YB-F7 of the present invention prevents and treats gaeumannomyces graminis pot experiment:
(1) test method: potted plant efficiency test carries out in Henan Academy of Agricultural Sciences greenhouse, by wheat seed through 1% hypochlorite disinfectant process 10min after, then use aseptic water washing 3 times, in 25 DEG C of thermostat containers, vernalization is for subsequent use to showing money or valuables one carries unintentionally.Adopt the inoculation of bacterium cake method, the total eclipse bacterium bacterium cake that 12 pieces of diameters are 6.0mm is placed in each little basin that sterilized soil is housed, be positioned over above Pathogen of Take-all bacterium cake by wheat seed after process and (every ferfas cake put one grained wheat seed), each treatment group arranges 4 basins and repeats.When emerging to height of seedling 5-8cm, carry out root irrigation.After one month, wheat seedling is extracted with root, with clear water, root is rinsed well, and observe statistics and to cure the disease effect (pot experiment schematic diagram refers to accompanying drawing 5, and statistics refers to table 4).
(2) process:
The antagonistic strain fermented liquid stoste adopted in following treating processes is cultivate the biocontrol microorganisms fermented liquid stoste obtained in embodiment 4.
Process 1: seed soaking.Antagonistic strain fermented liquid stoste seed soaking 3h;
Process 2: seed soaking.Antagonistic strain fermented liquid dilutes 5 times of seed soaking 3h;
Process 3: seed soaking.Antagonistic strain fermented liquid dilutes 10 times of seed soaking 3h;
Process 4: root irrigation.The former liquid irrigating root of antagonistic strain fermented liquid;
Process 5: root irrigation.Antagonistic strain fermented liquid dilutes 5 times and fills with root;
Process 6: root irrigation.Antagonistic strain fermented liquid dilutes 10 times and fills with root;
Process 7: clear water contrasts;
Process 8: medicament contrasts; 12.5% Silthiopham seed dressing.
(3) grade scale and drug effect method of calculation:
The concrete grade scale of take-all is:
0 grade: anosis;
1 grade: root system onset area accounts for 1% ~ 5%;
3 grades: root system onset area accounts for 6% ~ 20%;
5 grades: root system onset area accounts for 21% ~ 40%;
7 grades: root system onset area accounts for 41% ~ 60%;
9 grades: root system onset area accounts for more than 61%.
Table 4 biocontrol microorganisms YB-F7 prevents and treats take-all results from pot experiment test
| Process | Seedling rate (%) | Sickness rate (%) | Disease index (%) | Prevention effect (%) |
| Seed soaking stoste | 90.8 | 26 | 26 | 24 |
| To soak seed 5 times of stostes | 90 | 15 | 9 | 73.5 |
| To soak seed 10 times of stoste liquid | 86.7 | 32 | 27 | 21 |
| Former liquid irrigating root | 94.2 | 39 | 34 | 0 |
| Stoste 5 times fills with root | 79.2 | 13 | 8 | 76.5 |
| Stoste 10 times fills with root | 76.7 | 21 | 19 | 50 |
| Clear water contrasts | 93.3 | 50 | 34 | — |
| Total eclipse is dressed seed only | 53.3 | 7 | 7 | 79 |
Can be illustrated by table 4, after soaking seed and filling with root two kinds of method process, biocontrol microorganisms of the present invention has good prevention effect to take-all, and after especially fermented liquid dilutes 5 times, preventive effect and total eclipse net phase are worked as, and root irrigation mode prevention effect is higher than seed soaking.After all chemicals treatment, have certain influence to emerging, show as emerge late, but low all higher than other medicaments with the seedling rate after biocontrol microorganisms YB-F7 fermentation liquor treatment.
In sum, Methylotrophic genus bacillus (Bacillusmethylotrophicus) YB-F7 of the present invention all has remarkable fungistatic effect to gaeumannomyces graminis, sheath blight fungus and root-rot sickle-like bacteria; Therefore, Methylotrophic genus bacillus (Bacillusmethylotrophicus) YB-F7 of the present invention can be applied to preparation completely and prevents and treats in gaeumannomyces graminis disease microbial plant soil-borne diseases, banded sclerotial blight microbial roots of plants stem's disease or root-rot reaping hook microbial axis basal stem rot evil microbial inoculum.
Claims (4)
1. Methylotrophic genus bacillus (Bacillusmethylotrophicus) YB-F7, is characterized in that: this bacterial strain is preserved in China Committee for Culture Collection of Microorganisms's common micro-organisms center, and deposit number is CGMCCNO.10834.
2. Methylotrophic genus bacillus (Bacillusmethylotrophicus) YB-F7 according to claim 1, is characterized in that: the nucleotides sequence of described Methylotrophic genus bacillus YB-F7 is classified as SEQIDNO.1.
3. Methylotrophic genus bacillus (Bacillusmethylotrophicus) YB-F7 be used for biological control gaeumannomyces graminis disease microbial plant soil-borne diseases, application in banded sclerotial blight microbial roots of plants stem's disease or the microbial axis basal stem rot evil of root-rot reaping hook.
4. Methylotrophic genus bacillus (Bacillusmethylotrophicus) YB-F7 is for the preparation of the application prevented and treated in gaeumannomyces graminis disease microbial plant soil-borne diseases, banded sclerotial blight microbial roots of plants stem's disease or root-rot reaping hook microbial axis basal stem rot evil microbial inoculum.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201511026963.XA CN105543132B (en) | 2015-12-31 | 2015-12-31 | A kind of Methylotrophic bacillus YB-F7 and its application in controlling plant diseases |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201511026963.XA CN105543132B (en) | 2015-12-31 | 2015-12-31 | A kind of Methylotrophic bacillus YB-F7 and its application in controlling plant diseases |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN105543132A true CN105543132A (en) | 2016-05-04 |
| CN105543132B CN105543132B (en) | 2019-01-18 |
Family
ID=55822690
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201511026963.XA Active CN105543132B (en) | 2015-12-31 | 2015-12-31 | A kind of Methylotrophic bacillus YB-F7 and its application in controlling plant diseases |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN105543132B (en) |
Cited By (10)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN106857674A (en) * | 2017-02-09 | 2017-06-20 | 河北省科学院生物研究所 | The application of Methylotrophic bacillus BH21 and its microbial manure and preparation method |
| CN107760630A (en) * | 2017-11-24 | 2018-03-06 | 四川农业大学 | A kind of Methylotrophic bacillus B18 and its microbial inoculum and application |
| CN107973664A (en) * | 2017-12-11 | 2018-05-01 | 河南省农业科学院植物保护研究所 | Have biological organic fertilizer of disease-resistant growth-promoting functions and preparation method and application concurrently |
| CN108409479A (en) * | 2018-01-31 | 2018-08-17 | 河南省农业科学院植物保护研究所 | Nematicidal microbial organic fertilizer and its preparation method and application |
| CN108641989A (en) * | 2018-06-28 | 2018-10-12 | 山东农业大学 | One plant of Methylotrophic bacillus and its application |
| CN112136836A (en) * | 2020-09-17 | 2020-12-29 | 云南省农业科学院农业环境资源研究所 | Application of organic carbon fertilizer in preventing and treating crop soil-borne diseases and method for preventing and treating crop soil-borne diseases |
| CN112795506A (en) * | 2021-01-07 | 2021-05-14 | 河南省农业科学院植物保护研究所 | Bacillus belgii capable of preventing and treating root rot of salvia miltiorrhiza and application thereof |
| CN112795504A (en) * | 2021-01-07 | 2021-05-14 | 河南省农业科学院植物保护研究所 | Bacillus belgii capable of preventing and treating root rot of salvia miltiorrhiza and application thereof |
| CN112812993A (en) * | 2021-01-07 | 2021-05-18 | 河南省农业科学院植物保护研究所 | Bacillus belgii capable of preventing and treating root rot of salvia miltiorrhiza and application thereof |
| CN116083305A (en) * | 2022-12-30 | 2023-05-09 | 云南省微生物研究所 | Methylotrophic bacillus and biological compound fertilizer and application thereof |
Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102703363A (en) * | 2012-05-14 | 2012-10-03 | 大地绿源环保科技(北京)有限公司 | Bacillus methylotrophicus UTM401 and applications thereof |
| CN104342393A (en) * | 2014-10-31 | 2015-02-11 | 中国农业科学院农产品加工研究所 | Bacillus methylotrophicus and application thereof on aspect of preventing and controlling fusarium graminearum |
| CN105018386A (en) * | 2015-07-31 | 2015-11-04 | 长江大学 | Bacillus methylotrophicus and application thereof |
-
2015
- 2015-12-31 CN CN201511026963.XA patent/CN105543132B/en active Active
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102703363A (en) * | 2012-05-14 | 2012-10-03 | 大地绿源环保科技(北京)有限公司 | Bacillus methylotrophicus UTM401 and applications thereof |
| CN104342393A (en) * | 2014-10-31 | 2015-02-11 | 中国农业科学院农产品加工研究所 | Bacillus methylotrophicus and application thereof on aspect of preventing and controlling fusarium graminearum |
| CN105018386A (en) * | 2015-07-31 | 2015-11-04 | 长江大学 | Bacillus methylotrophicus and application thereof |
Non-Patent Citations (4)
| Title |
|---|
| FUHONG XIE ET AL.: "Purification and characterization of a novel α-amylase from", 《PROCESS BIOCHEMISTRY》 * |
| MUNUSAMY MADHAIYAN ET AL.: "Bacillus methylotrophicus sp. nov., a methanolutilizing,", 《INTERNATIONAL JOURNAL OF SYSTEMATIC AND EVOLUTIONARY MICROBIOLOGY》 * |
| 张帆等: "小麦全蚀病菌拮抗菌株FY1的筛选和鉴定", 《河南农业科学》 * |
| 成卓敏: "《粮食安全与植保科技创新》", 30 September 2009 * |
Cited By (17)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN106857674B (en) * | 2017-02-09 | 2019-02-26 | 河北省科学院生物研究所 | The application of Methylotrophic bacillus BH21 and its microbial manure and preparation method |
| CN106857674A (en) * | 2017-02-09 | 2017-06-20 | 河北省科学院生物研究所 | The application of Methylotrophic bacillus BH21 and its microbial manure and preparation method |
| CN107760630A (en) * | 2017-11-24 | 2018-03-06 | 四川农业大学 | A kind of Methylotrophic bacillus B18 and its microbial inoculum and application |
| CN107760630B (en) * | 2017-11-24 | 2021-02-02 | 四川农业大学 | Bacillus methylotrophicus B18, microbial inoculum and application thereof |
| CN107973664A (en) * | 2017-12-11 | 2018-05-01 | 河南省农业科学院植物保护研究所 | Have biological organic fertilizer of disease-resistant growth-promoting functions and preparation method and application concurrently |
| CN108409479A (en) * | 2018-01-31 | 2018-08-17 | 河南省农业科学院植物保护研究所 | Nematicidal microbial organic fertilizer and its preparation method and application |
| CN108641989B (en) * | 2018-06-28 | 2021-06-15 | 山东农业大学 | Bacillus methylotrophicus and application thereof |
| CN108641989A (en) * | 2018-06-28 | 2018-10-12 | 山东农业大学 | One plant of Methylotrophic bacillus and its application |
| CN112136836B (en) * | 2020-09-17 | 2021-11-23 | 云南省农业科学院农业环境资源研究所 | Application of organic carbon fertilizer in preventing and treating crop soil-borne diseases and method for preventing and treating crop soil-borne diseases |
| CN112136836A (en) * | 2020-09-17 | 2020-12-29 | 云南省农业科学院农业环境资源研究所 | Application of organic carbon fertilizer in preventing and treating crop soil-borne diseases and method for preventing and treating crop soil-borne diseases |
| CN112795504A (en) * | 2021-01-07 | 2021-05-14 | 河南省农业科学院植物保护研究所 | Bacillus belgii capable of preventing and treating root rot of salvia miltiorrhiza and application thereof |
| CN112812993A (en) * | 2021-01-07 | 2021-05-18 | 河南省农业科学院植物保护研究所 | Bacillus belgii capable of preventing and treating root rot of salvia miltiorrhiza and application thereof |
| CN112795506A (en) * | 2021-01-07 | 2021-05-14 | 河南省农业科学院植物保护研究所 | Bacillus belgii capable of preventing and treating root rot of salvia miltiorrhiza and application thereof |
| CN112795506B (en) * | 2021-01-07 | 2022-08-19 | 河南省农业科学院植物保护研究所 | Bacillus beleisi capable of preventing and treating root rot of salvia miltiorrhiza and application thereof |
| CN112795504B (en) * | 2021-01-07 | 2022-08-19 | 河南省农业科学院植物保护研究所 | Bacillus belgii capable of preventing and treating root rot of salvia miltiorrhiza and application thereof |
| CN112812993B (en) * | 2021-01-07 | 2022-08-19 | 河南省农业科学院植物保护研究所 | Bacillus belgii capable of preventing and treating root rot of salvia miltiorrhiza and application thereof |
| CN116083305A (en) * | 2022-12-30 | 2023-05-09 | 云南省微生物研究所 | Methylotrophic bacillus and biological compound fertilizer and application thereof |
Also Published As
| Publication number | Publication date |
|---|---|
| CN105543132B (en) | 2019-01-18 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN105886428B (en) | One plant of Streptomycesalbidoflhaving and its application in microbial manure | |
| CN105543132B (en) | A kind of Methylotrophic bacillus YB-F7 and its application in controlling plant diseases | |
| CN105296381B (en) | One bacillus subtilis CYY-25 and its application | |
| CN102433282B (en) | Bacillus subtilis NB12, as well as culture method and application thereof | |
| CN104164394B (en) | The bacterial strain of one strain antagonistic phytopathogen and application thereof | |
| CN101822272B (en) | Streptomyces griseoflavus and application thereof in biological prevention and control of plant diseases | |
| CN101851597B (en) | Streptomyces griseoflavus for resisting alfalfa diseases and screening method thereof | |
| CN105441366B (en) | Methylotrophic bacillus ZBL-1 is applied in preventing cotton verticillium wilt | |
| CN106119155B (en) | For preventing and treating the bacterial strain WXX-2 and microbial inoculum of peanut root rot | |
| CN105039220B (en) | Methylotrophic bacillus AR3, bacillus subtilis AR4, bacillus amyloliquefaciens AR10 and application | |
| CN102533593A (en) | Burkholderia cepacia SD7 and culturing method and application thereof | |
| CN111073825B (en) | Bacterium with plant soil-borne disease resistance effect and application thereof | |
| CN110205248B (en) | Method for promoting plant growth by jointly inoculating AM and DSE fungi and microbial agent used by method | |
| CN103074271A (en) | Bacillus subtilis YB-05, microbial preparation thereof, and application of Bacillus subtilis YB-05 or microbial preparation | |
| CN110423707A (en) | Bacillus subtilis strain NH-6 and its application | |
| CN106399152A (en) | Streptomyces having inhibitory effect on phytophthorasojae root rot fungi, biocontrol preparation and preparation method of biocontrol preparation | |
| CN105132296B (en) | A kind of hook-shaped trichoderma strain and its application | |
| CN105238723B (en) | A kind of bacillus amyloliquefaciens and its microbial bacterial agent of prevention crop verticillium wilt | |
| CN110468057A (en) | One plants endogenetic disk both-end hair spore category fungi M7SB 41 and its application | |
| CN106701631A (en) | Strain of streptomyces roseoflavus and application thereof | |
| CN108676745A (en) | Solution xylose lysine bacillus LX024 and its application of one plant of prevention wheat base rot disease and head blight | |
| CN103146609B (en) | Pseudomonas fluorescens and method for preventing phytophthora capsici thereby | |
| CN109749953B (en) | Bacillus cereus, microbial inoculum and preparation method and application thereof | |
| CN110317747A (en) | A kind of bacillus amyloliquefaciens JT68 and its application in prevention and treatment tea anthracnose | |
| CN107502570B (en) | One plant of Biocontrol Bacillus subtilis BJ-1 and its application |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| GR01 | Patent grant | ||
| GR01 | Patent grant |