CN108130358A - To the research method of pseudomonas aeruginosa-aspergillus fumigatus mixed biologic envelope inhibiting effect - Google Patents
To the research method of pseudomonas aeruginosa-aspergillus fumigatus mixed biologic envelope inhibiting effect Download PDFInfo
- Publication number
- CN108130358A CN108130358A CN201711388694.0A CN201711388694A CN108130358A CN 108130358 A CN108130358 A CN 108130358A CN 201711388694 A CN201711388694 A CN 201711388694A CN 108130358 A CN108130358 A CN 108130358A
- Authority
- CN
- China
- Prior art keywords
- aspergillus fumigatus
- aeruginosa
- pseudomonas aeruginosa
- group
- 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.)
- Pending
Links
Classifications
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
- C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
- C12Q1/02—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving viable microorganisms
- C12Q1/04—Determining presence or kind of microorganism; Use of selective media for testing antibiotics or bacteriocides; Compositions containing a chemical indicator therefor
- C12Q1/06—Quantitative determination
Landscapes
- Chemical & Material Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Health & Medical Sciences (AREA)
- Organic Chemistry (AREA)
- Engineering & Computer Science (AREA)
- Zoology (AREA)
- Wood Science & Technology (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- Analytical Chemistry (AREA)
- Toxicology (AREA)
- Immunology (AREA)
- Microbiology (AREA)
- Molecular Biology (AREA)
- Biophysics (AREA)
- Physics & Mathematics (AREA)
- Biotechnology (AREA)
- Biochemistry (AREA)
- Bioinformatics & Cheminformatics (AREA)
- General Engineering & Computer Science (AREA)
- General Health & Medical Sciences (AREA)
- Genetics & Genomics (AREA)
- Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
Abstract
The present invention provides a kind of close research methods to pseudomonas aeruginosa aspergillus fumigatus mixed biologic envelope inhibiting effect, include the following steps：The preparation of aspergillus fumigatus spores suspension；Pseudomonas aeruginosa wild strain and its defect bacterial strain bacterium solution prepare picking pseudomonas aeruginosa wild strain single bacterium colony and P. aeruginosa strain density sensing defect bacterial strain single bacterium colony, are inoculated in culture medium respectively, then dilute；External static two kinds of mixed biologics are divided into 6 groups by the foundation experiment of membrane modle, every group is inoculated in tissue culture plate, wherein respectively place circular glass cell climbing sheet in advance per hole, wild type P. aeruginosa bacterium solution and P. aeruginosa strain density sensing defect bacterium solution are separately added into respective sets again, constant-temperature incubation；After being formed for biofilms different in above-mentioned steps, quantitative analysis is carried out.The present invention specifies that P. aeruginosa strain density induction system has significant inhibiting effect to the formation of pseudomonas aeruginosa-aspergillus fumigatus mixed biologic envelope.
Description
Technical field
The present invention relates to a kind of methods for inhibiting biofilm, and in particular to a kind of external inhibition pseudomonas aeruginosa-cigarette
The research method of Aspergillus mixed biologic envelope, belongs to biological correlative technology field.
Background technology
Pseudomonas aeruginosa and aspergillus fumigatus are bacteriums and fungi more typical in nature, and are caused for more typical condition
Germ.Either the secondary intrapulmonary infection of chronic obstructive pulmonary disease or branch expand the common field planting bacterium of intrapulmonary, and also or incidence shows note raising in recent years
Pulmonary tuberculosis merge bacterial fungus infection, pseudomonas aeruginosa and aspergillus fumigatus are the common mushroom of culture.Bacterium and true
Bacterium mainly exists in people's tumor growth in a manner of forming biofilm, and mixed infection can then form mixed biologic envelope and be allowed to send out
Wave more powerful drug resistance.Less is reported for the research of mixed biologic envelope both at home and abroad at present, domestic and foreign literature is about copper
The research of green pseudomonad and aspergillus fumigatus is only limitted to research of the pseudomonas aeruginosa for the inhibiting effect of aspergillus fumigatus, and does not have
Have and be deep into the research field of mixed biologic envelope that the two is formed, this has just given the much room of the research.
Density induction system refers to that by monitoring the cell density of its group, spy is adjusted using own signal molecule for bacterium
Fixed gene expression to ensure the transport of nutriment and the discharge of waste in biofilm, avoids bacterial overgrowth and makes
Lack into space and nutriment.Density induction system includes the adjustment effect of bacterium：Biofilm formation, bioluminescence,
The adjusting of separate gene, generation of antibiotic etc..
Pseudomonas aeruginosa-aspergillus fumigatus mixed biologic envelope is ground using the density induction system of pseudomonas aeruginosa
Study carefully and still belong to blank.
Invention content
The object of the present invention is to provide a kind of external sides for inhibiting pseudomonas aeruginosa-aspergillus fumigatus mixed biologic envelope
Method, to make up the deficiencies in the prior art.
The density induction system of pseudomonas aeruginosa mainly includes：Tri- systems of las, rhl, pqs, wherein las systems are lived
Change the transcription of a large amount of regulatory factors such as rhlR.rhlI, thus critical role is accounted in its adjusting in verdigris virulence determinant.Its
Middle las is made of lasR, lasI gene, and rhl is made of rhlR, rhlI gene.
In order to achieve the above objectives, the specific technical solution taken of the present invention is：
A kind of density induction system is to the research side of pseudomonas aeruginosa-aspergillus fumigatus mixed biologic envelope inhibiting effect
Method includes the following steps：
(1) preparation of aspergillus fumigatus spores suspension；
(2) preparation of pseudomonas aeruginosa wild strain and its defect bacterial strain bacterium solution：Picking pseudomonas aeruginosa wild mushroom
Strain single bacterium colony and P. aeruginosa strain density sensing defect bacterial strain single bacterium colony, are inoculated in RPMI-1640 culture mediums, then dilute respectively
It releases；
(3) external static two kinds of mixed biologics are divided into 6 groups by the foundation experiment of membrane modle, are aspergillus fumigatus-wild successively
Type pseudomonas aeruginosa group (Af+PAO1), aspergillus fumigatus-P. aeruginosa strain density sensing defect bacterium group (A.f+P.a
LasI/rhlI), aspergillus fumigatus group (A.f), wild type pseudomonas aeruginosa group (PAO1), the sensing of P. aeruginosa strain density lack
Bacterium group (P.a lasI/rhlI) is fallen into, RPMI-1640 culture mediums are inoculated in tissue culture plate as blank control group, every group, wherein
The respectively placement circular glass cell climbing sheet, then by wild type P. aeruginosa bacterium solution and P. aeruginosa strain density sense in advance per hole
Defect bacterium solution is answered to be separately added into respective sets, constant-temperature incubation；
(4) after being formed for biofilms different in above-mentioned steps (3), quantitative analysis is carried out.
Further, the step (1) is specially：Tested aspergillus fumigatus bacterial strain is seeded to potato dextrose agar slant training
Base is supported, collects spore, RPMI-1640 culture mediums are configured；The spore liquid being collected into is added in the RPMI-1640 culture mediums,
Spore concentration is adjusted with blood cell counting plate, as establishing external mixed biologic by the spore suspension of membrane modle.
Further, in the step (2) after dilution, final concentration of the 1 × 10 of above two bacterium^8CFU/mL。
Further, in the step (4), quantitative analysis is carried out using 4 crystal violet staining assays.
A kind of external method for inhibiting pseudomonas aeruginosa-aspergillus fumigatus mixed biologic envelope, this method are false using verdigris
The density induction system of monad inhibits the biofilm.
Further, above-mentioned density induction system is lasI/rhlI type density induction systems.
Advantages of the present invention and technique effect：
The present invention specifies P. aeruginosa strain density induction system to pseudomonas aeruginosa-aspergillus fumigatus mixed biologic
The formation of envelope has significant inhibiting effect.Can OD values and just be measured by microplate reader by crystal violet native staining method respectively
The morphosis of microscope Microscopic observation each group mixed biologic envelope is put to obtain P. aeruginosa strain density induction system to copper
Conclusion of the formation with significant inhibiting effect of green pseudomonad-aspergillus fumigatus mixed biologic envelope.The present invention utilizes density
Induction system is inhibited to the formation of pseudomonas aeruginosa-aspergillus fumigatus mixed biologic envelope.For the treatment of infection
And the application of associated antibiotic has positive reference and directive function.
This experimental method can also be used for other kinds of density sensing defect P. aeruginosa bacterial strain and aspergillus fumigatus institute shape
Into mixed biologic envelope research, observe its comparison with wild type pseudomonas aeruginosa-aspergillus fumigatus mixed biologic envelope,
So as to which the density induction system for finding pseudomonas aeruginosa has wild type pseudomonas aeruginosa-aspergillus fumigatus mixed biologic envelope
There is inhibiting effect.Also other bacteriums-other Mycophytas are mixed available for the density induction system of the other bacteriums of observational study
Biofilm whether there is the research of inhibiting effect.
Description of the drawings
Fig. 1 is the comparative analysis figure of the other absorbance value of each group in the present invention.
Fig. 2 is each group electron microscope in the present invention.
Specific embodiment
It is explained further and illustrates below by way of specific embodiment and with reference to attached drawing to the present invention.
Embodiment 1：
A kind of density induction system for verifying pseudomonas aeruginosa to inhibiting pseudomonas aeruginosa-aspergillus fumigatus to mix in vitro
The method for closing biofilm, includes the following steps：
(1) the tested aspergillus fumigatus bacterial strain for preparing of aspergillus fumigatus spores suspension is seeded to potato dextrose agar slant culture
Base, 37 DEG C of constant temperature recoveries 72 hours rinse potato dextrose agar slant medium slant with 8mL sterile pure waters and collect spore
Son.The RPMI-1640 culture mediums that pH is 7.0 after configuration 3- (N- morpholinyls) propane sulfonic acid -4- N-morpholinyls buffering.It will collect
To about 8mL spore liquids be added in the about 30mL RPMI-1640 culture mediums, it is 1 to adjust spore concentration with blood cell counting plate
×10^6CFU/mL as external mixed biologic is established by the spore suspension of membrane modle, matches for 16 hours before biofilm mixing
System；
(2) pseudomonas aeruginosa wild strain and its defect bacterial strain bacterium solution prepare picking pseudomonas aeruginosa wild strain
Single bacterium colony (such as icon is denoted as 35) and P. aeruginosa strain density sensing defect bacterial strain single bacterium colony (such as icon is denoted as 42), connect respectively
Kind pH after 20mL 3- (N- morpholinyls) propane sulfonic acid -4- N-morpholinyls buffering is 37 DEG C in 7.0 RPMI-1640 culture mediums
After constant-temperature table is incubated 20-24 hours (220 revs/min of rotating speed), after being buffered with 3- (N- morpholinyls) propane sulfonic acid -4- N-morpholinyls
The RPMI-1640 culture mediums that pH is 7.0 dilute 10 times, make final concentration of the 1 × 10 of both bacteriums^8CFU/mL；
(3) external static two kinds of mixed biologics are divided into 6 groups by the foundation experiment of membrane modle, are aspergillus fumigatus-wild successively
Type pseudomonas aeruginosa group (Af+PAO1), aspergillus fumigatus-P. aeruginosa strain density sensing defect bacterium group (A.f+
P.alasI/rhlI), aspergillus fumigatus group (A.f), wild type pseudomonas aeruginosa group (PAO1), the sensing of P. aeruginosa strain density
Defect bacterium group (P.a lasI/rhlI), RPMI-1640 culture mediums are as blank control group.It is inoculated in 24 hole cell culture of sterilizing
Plate, every group of 3 multiple holes, each boreliquid total amount are 1550 μ L, and aspergillus fumigatus liquid measure and wild type pseudomonas aeruginosa and verdigris are false
The ratio between unit cell strain density sensing defect bacterium solution amount is 30：1.The sterilizing for respectively placing one piece of a diameter of 13mm in advance per hole is round
Glass cell climbing sheet.The aspergillus fumigatus spores suspension prepared and 1640 liquid are respectively positioned over containing aspergillus fumigatus before 16h
And in the hole of blank group, and it is incubated in 37 DEG C of insulating boxs.By wild type P. aeruginosa bacterium solution and pseudomonas aeruginosa after 16h
Density sensing defect bacterium solution is separately added into respective sets.In 37 DEG C of constant-temperature incubations for 24 hours；
Quantitative analysis after (4) 4 crystal violet staining assays form different biofilms mixes life for two kinds according to the method described above
After object envelope builds film for 24 hours, gently wipe the flcating germ of each hole surface away with sterilized cotton swabs, then discard culture medium, use sterile pure water
Each hole is gently rinsed 3 times to remove flcating germ, and pumps remaining sterile pure water in each hole with disposable 2mL syringes as possible.In
Each hole adds in 2% glutaraldehyde solution 1mL room temperatures and fixes 2 hours.Drying at room temperature after exhaustion glutaraldehyde.Each hole adds in 0.5% crystallization
Purple 1mL is dyed 15 minutes.After 15 minutes remaining crystal violet solution in each hole is pumped with disposable 2mL syringes.Drying at room temperature.Room
1mL95% ethanol solutions decoloration 10min is added in after temperature is dry per hole.It draws 100 μ L eluents and adds in 96 orifice plates in each hole.Microplate reader
590nm wavelength measures each hole absorbance value (OD 590).Every group sets 3 multiple holes to take its average value respectively every time, and experiment is independent
It is repeated 3 times.
2 interpretation of result of embodiment：
As shown in Figure 1, this group of 1-6 group of data is corresponding in turn to：1 aspergillus fumigatus-wild type pseudomonas aeruginosa group；2 cigarettes are bent
Mould-P. aeruginosa strain density sensing defect bacterium group；3 aspergillus fumigatus groups；4 wild type pseudomonas aeruginosa groups；5 verdigris are false single
Born of the same parents' strain density senses defect bacterium group；6RPMI-1640 culture mediums are as blank control group.Thus group data can be seen that group 1 compares group
2 mean OD value significantly reduces and difference is statistically significant (carries out data analysis, metering using SPSS17.0 statistics softwares
Data represents that more comparison among groups one-way analysis of variances and SNK are examined, 0.05 differences of P ﹤ with mean ± standard deviation (X- ± S)
It is statistically significant).It illustrates, density induction system has significantly pseudomonas aeruginosa-aspergillus fumigatus mixed biologic envelope
Inhibiting effect.
As shown in Fig. 2, wherein, (1) is aspergillus fumigatus-wild type pseudomonas aeruginosa group, (2) are aspergillus fumigatus-verdigris
Pseudomonad density senses defect bacterium group, and (3) are aspergillus fumigatus group, and (4) are wild type pseudomonas aeruginosa group, and (5) are verdigris
Pseudomonad density senses defect bacterium group, and (6) are RPMI-1640 blank control groups；(7) it is amplified aspergillus fumigatus-wild
Type pseudomonas aeruginosa group, (8) sense defect bacterium group for amplified aspergillus fumigatus-P. aeruginosa strain density, and (9) are amplification
Aspergillus fumigatus group afterwards, (10) are amplified wild type pseudomonas aeruginosa group, and (11) are amplified pseudomonas aeruginosa
Density senses defect bacterium group, and (12) are amplified RPMI-1640 blank control groups.
It can be seen from the figure that aspergillus fumigatus-mixed biologic envelope amount caused by wild type pseudomonas aeruginosa group (causes
Close degree) it is significantly (finer and close than aspergillus fumigatus-mixed biologic envelope amount caused by P. aeruginosa strain density sensing defect bacterium group
Degree) it reduces.It can equally illustrate formation of the density induction system to pseudomonas aeruginosa-aspergillus fumigatus mixed biologic envelope
It is inhibited.
Claims (4)
1. a kind of research method to pseudomonas aeruginosa-aspergillus fumigatus mixed biologic envelope inhibiting effect, which is characterized in that should
Research method includes the following steps：
(1) preparation of aspergillus fumigatus spores suspension；
(2) pseudomonas aeruginosa wild strain and its defect bacterial strain bacterium solution prepare picking pseudomonas aeruginosa wild strain single bacterium
It falls and P. aeruginosa strain density senses defect bacterial strain single bacterium colony, be inoculated in respectively in RPMI-1640 culture mediums, then dilute；
(3) external static two kinds of mixed biologics are divided into 6 groups by the foundation experiment of membrane modle, are aspergillus fumigatus-wild type copper successively
Green pseudomonad group (Af+PAO1), aspergillus fumigatus-P. aeruginosa strain density sensing defect bacterium group (A.f+P.a lasI/
RhlI), aspergillus fumigatus group (A.f), wild type pseudomonas aeruginosa group (PAO1), P. aeruginosa strain density sensing defect bacterium group
(P.alasI/rhlI), RPMI-1640 culture mediums are inoculated in tissue culture plate as blank control group, every group, wherein per hole
It is each in advance to place circular glass cell climbing sheet, then wild type P. aeruginosa bacterium solution and P. aeruginosa strain density are sensed into defect
Bacterium solution is separately added into respective sets, constant-temperature incubation；
(4) after being formed for biofilms different in above-mentioned steps (3), quantitative analysis is carried out.
2. research method as described in claim 1, which is characterized in that the step (1) is specially：Tested aspergillus fumigatus bacterial strain connects
Kind collects spore, RPMI-1640 culture mediums is configured to potato dextrose agar slant culture medium；The spore liquid that will be collected into
It is added in the RPMI-1640 culture mediums, spore concentration is adjusted with blood cell counting plate, as establishing external mixed biologic envelope
The spore suspension of model.
3. research method as described in claim 1, which is characterized in that in the step (2) after dilution, above two is thin
Final concentration of the 1 × 10 of bacterium^8CFU/mL。
4. research method as described in claim 1, which is characterized in that in the step (4), using 4 crystal violet staining assays into
Row quantitative analysis.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201711388694.0A CN108130358A (en) | 2017-12-21 | 2017-12-21 | To the research method of pseudomonas aeruginosa-aspergillus fumigatus mixed biologic envelope inhibiting effect |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201711388694.0A CN108130358A (en) | 2017-12-21 | 2017-12-21 | To the research method of pseudomonas aeruginosa-aspergillus fumigatus mixed biologic envelope inhibiting effect |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN108130358A true CN108130358A (en) | 2018-06-08 |
Family
ID=62390981
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201711388694.0A Pending CN108130358A (en) | 2017-12-21 | 2017-12-21 | To the research method of pseudomonas aeruginosa-aspergillus fumigatus mixed biologic envelope inhibiting effect |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN108130358A (en) |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JP2021048779A (en) * | 2019-09-24 | 2021-04-01 | 小林製薬株式会社 | Dark stain forming method |
Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO1998057618A1 (en) * | 1997-06-18 | 1998-12-23 | The Research And Development Institute, Inc. | Methods and compositions for controlling biofilm development |
| WO2001018248A2 (en) * | 1999-09-03 | 2001-03-15 | The University Of Iowa Research Foundation | Quorum sensing signaling in bacteria |
| CN101001635A (en) * | 2003-12-04 | 2007-07-18 | 生物膜策略公司 | Methods and compositions for preventing biofilm formation, reducing existing biofilms, and for reducing populations of bacteria |
| CN102482244A (en) * | 2009-07-03 | 2012-05-30 | 内盖夫国家生物技术有限公司 | Covalent inhibition of bacterial quorum sensing |
-
2017
- 2017-12-21 CN CN201711388694.0A patent/CN108130358A/en active Pending
Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO1998057618A1 (en) * | 1997-06-18 | 1998-12-23 | The Research And Development Institute, Inc. | Methods and compositions for controlling biofilm development |
| WO2001018248A2 (en) * | 1999-09-03 | 2001-03-15 | The University Of Iowa Research Foundation | Quorum sensing signaling in bacteria |
| CN101001635A (en) * | 2003-12-04 | 2007-07-18 | 生物膜策略公司 | Methods and compositions for preventing biofilm formation, reducing existing biofilms, and for reducing populations of bacteria |
| CN102482244A (en) * | 2009-07-03 | 2012-05-30 | 内盖夫国家生物技术有限公司 | Covalent inhibition of bacterial quorum sensing |
Non-Patent Citations (3)
| Title |
|---|
| EILIDH MOWAT 等: "Pseudomonas aeruginosa and their small diffusible extracellular molecules inhibit Aspergillus fumigatus biofilm formation", 《FEMS MICROBIOL LETT》 * |
| GABRIELE SASS 等: "Studies of Pseudomonas aeruginosa Mutants Indicate Pyoverdine as the Central Factor in Inhibition of Aspergillus fumigatus Biofilm", 《J BACTERIOL》 * |
| 唐佩 等: "铜绿假单胞菌生物膜形成调控机制的研究进展", 《浙江预防医学》 * |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JP2021048779A (en) * | 2019-09-24 | 2021-04-01 | 小林製薬株式会社 | Dark stain forming method |
| JP7370203B2 (en) | 2019-09-24 | 2023-10-27 | 小林製薬株式会社 | How to form blackheads |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN104745672B (en) | A kind of method of Rapid identification tobacco black shank resistance | |
| CN104611254A (en) | BaciIllus polymyxa KN-03 and culturing method and use thereof | |
| CN107828665A (en) | A kind of isolation and purification method of purpleback murdannia herb endogenetic fungus and its application | |
| CN107475172A (en) | A kind of method for obtaining Geobacillus stearothermophilus gemma | |
| CN110885776A (en) | Cellulose streptomyces fermentation extract and application thereof | |
| CN108130358A (en) | To the research method of pseudomonas aeruginosa-aspergillus fumigatus mixed biologic envelope inhibiting effect | |
| Kelsey | The testing of sterilizers: 2 Thermophilic spore papers | |
| CN104611255B (en) | One plant height coherency Pediococcus pentosaceus and its application in purifying water body in vibrio parahemolyticus | |
| CN110257316A (en) | A kind of plant anthrax bacteria conidium rapid separation and purification method | |
| CN112553293A (en) | Method for rapidly evaluating in-vitro antagonistic activity of helicobacter pylori | |
| CN112501242A (en) | Method for screening biocontrol bacteria for tobacco bacterial wilt by using living biological test system | |
| CN106967669A (en) | A kind of quick, a large amount of acquisition conidial cultural method of cabbage heart anthrax bacteria | |
| CN109266560A (en) | A kind of Culture medium for Alternaria alternata containing weeds | |
| CN101750478B (en) | In-vitro verification method for apple rot disease resistance | |
| CN109169713A (en) | A kind of Bei Laisi bacillus ZSY-1 wettable powder and preparation method and application | |
| CN111548943B (en) | Oxytropis ochrantha endophytic fungus and application thereof | |
| CN114774293A (en) | Stachybotrys botrytis HN17496 strain, biocontrol microbial inoculum and preparation method and application thereof | |
| CN114703094A (en) | Bacillus pseudomycoides and application thereof in preparation of plant pathogenic bacteria bacteriostat | |
| US20210062237A1 (en) | Device for detecting tomato pathogenic fungus and detection method using the same | |
| CN109497050B (en) | Paenibacillus polymyxa fermentation product and preparation method and application thereof | |
| CN104745522B (en) | A kind of method for inducing silk spore fungi spore | |
| Catarino et al. | Morphological aspects and effect of carbon sources in the physiology of Fusarium oxysporum f. sp. passiflorae | |
| CN113416656B (en) | Geotrichum japonicum endophytic fungus strain HUBU0122 and application thereof | |
| CN113832039B (en) | Industrial cannabis endophytic fungus with antifungal activity and application thereof | |
| CN104120084A (en) | Metarhizium flavoviride MFYY090714 and applications 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 | ||
| RJ01 | Rejection of invention patent application after publication | ||
| RJ01 | Rejection of invention patent application after publication |
Application publication date: 20180608 |