CN102191185B - Biological control endophytic fungi - Aspergillus restrictus - Google Patents
Biological control endophytic fungi - Aspergillus restrictus Download PDFInfo
- Publication number
- CN102191185B CN102191185B CN201110088205A CN201110088205A CN102191185B CN 102191185 B CN102191185 B CN 102191185B CN 201110088205 A CN201110088205 A CN 201110088205A CN 201110088205 A CN201110088205 A CN 201110088205A CN 102191185 B CN102191185 B CN 102191185B
- Authority
- CN
- China
- Prior art keywords
- aspergillus restrictus
- garlic
- endophytic fungi
- bacterium
- 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.)
- Expired - Fee Related
Links
Landscapes
- Agricultural Chemicals And Associated Chemicals (AREA)
- Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)
Abstract
The invention relates to the microbial technology field. The garlic endophytic fungi are obtained from a garlic living body with endophytic fungi microbe separation technology, wherein the garlic living body is obtained at a vegetable garden in the suburb of Hangzhou city Zhejiang province. Through the microbe taxology identification, the garlic endophytic fungi are named as Aspergillus restrictus 28. The bacterial strain is preserved at China General Microbiological Culture Collection Center with a preservation date December 31st 2010 and a preservation registry number CGMCC No. 4507. The most substantial characteristic of the endophytic fungi is that through liquid fermentation the bacterial strain can produce an active metabolite which has an inhibition effect on pathogen of plant fungous disease such as Rhizoctonia solani, Thanatephorus cucumeris and the like. The bacterial strain is an important microbial resource.
Description
Technical field
The present invention relates to microbial technology field, particularly relate to the biological and ecological methods to prevent plant disease, pests, and erosion endogenetic fungus---the cultivation of Aspergillus restrictus bacterium (Aspergillus restrictus) and the application of meta-bolites thereof.
Background technology
All have a large amount of endogenetic fungus in the plant materials, endogenetic fungus is found in herbage the earliest, in plants such as xylophyta, herbage, gramineae plant, liver moss, lichens, marine alga, comes to light again afterwards.Research so far shows, plant endogenesis epiphyte can strengthen the host disease resistance, improve the yield-power, degeneration-resistant pest-resistant etc. of plant.Bringing into play these effects is because endogenetic fungus is bred in plant materials and grown, and generation has bioactive secondary metabolite in vivo.Therefore, endogenetic fungus possibly become potential biocontrol agent in the biological control, has important purposes aspect the ecological agriculture and the biological pesticide development.Plant endogenesis epiphyte has caused microbiologist, ecologist, plant protection scholar's extensive interest in recent years, becomes the focus of domestic and international research gradually.The research and development of microbial source biological chemistry agricultural chemicals at present is very active in China, but Microbial resources and application potential thereof still do not fully played, and plant endogenesis epiphyte promptly is such one type of untapped emerging Microbial resources.
At present, mainly adopt chemical pesticide to kill pathogenic bacteria, but chemical pesticide still is difficult to effective solution so far to people and animals' toxic side effect and residue problem to the control of Plant diseases.Utilize the active substance of the disease-resistant fungal pathogens of endogenetic fungus generation to carry out the biological control of plant pathogenic fungi disease, then have the cycle weak point, be easy to research, be convenient to advantages such as production and low toxicity.
The present invention is separated to a strain biological and ecological methods to prevent plant disease, pests, and erosion endogenetic fungus from garlic, and this strain fermentation meta-bolites has been carried out bacteriostatic action research.This invention provides good starting strain for the further development and the exploitation of microbial pesticide.
Summary of the invention
The objective of the invention is the dangerous and less deficiency of microbial pesticide kind, a kind of endogenetic fungus that from garlic, separates-Aspergillus restrictus bacterium (Aspergillus restrictus) is provided to chemical pesticide; Another object of the present invention provides the method for utilizing this bacterial strain meta-bolites control fungal diseases of plants.
The separation of Aspergillus restrictus bacterium of the present invention, screening and evaluation:
Be in vegetable garden, Zhejiang Hangzhou countryside, to gather garlic, obtain and preservation through steps such as separation, purifying, cultivation, fermentation and determinations of activity in April, 2009; Be accredited as Aspergillus restrictus bacterium (Aspergillus restrictus) through microbial taxonomy, name and be Aspergillus restrictus bacterium (Aspergillus restrictus) 28.This bacterial strain is in China Committee for Culture Collection of Microorganisms's common micro-organisms center preservation, preservation date on December 31st, 2010, preservation registration number CGMCCNo.4507.
Aspergillus restrictus bacterium 28, its solid culture is characterized as: bacterium colony 28 ℃ of cultivations on the PDA flat board extended to diameter 9cm in 5-7 days, and initial stage white is sparse, and mycelia crawls to grow along media surface, and the back forms grey, and reverse side is colourless, and the mycelia tool is separated, branch.The microscopic morphology characteristic: the conidiophore ultimate swelling is head; Conidium is light grey, and is subsphaeroidal.
Aspergillus restrictus bacterium 28 of the present invention can obtain the inhibited meta-bolites of part fungal diseases of plants pathogenic bacteria through fermented extracted.
The object of the invention realizes through following technical scheme:
A kind of preparation method who prevents and treats fungal diseases of plants microbial metabolites such as rice sheath blight disease or cucumber rhizoctonia rot, carry out according to the following steps:
(1) activation culture of bacterial classification: will separate Aspergillus restrictus 28 bacterial classifications of preserving and move on the potato glucose PDA solid plate, and under 28 ℃+1 ℃ condition, cultivate, and cover whole petridish to mycelia, using diameter is that the punching of 4mm punch tool obtains the bacterium cake, supplies inoculation usefulness;
(2) fermentation culture: fermentation culture is to contain soybean cake powder 20g among every 1000mL, Semen Maydis powder 15g, Zulkovsky starch 20g, wheat bran 10g, peptone 8g, NH
4NO
33g, (NH
4)
2SO
45g, CaCO
35g, MgSO
41g, the bottled fermentation culture 60mL of every 300mL triangle; Prepare back 121 ℃ the sterilization 20min, to be cooled to 40 ℃ under aseptic condition picking 1 diameter be 4mm bacterium cake, under 28 ℃ ± 1 ℃ condition, 180 rev/mins of shaking speed, fermentation culture 7 days;
(3) fermented product extracts: fermentation finishes, and centrifugal collection fermented supernatant fluid is used conventional organic solvent extraction, behind vacuum concentration, obtains medicinal extract, is used for bacteriostatic activity and measures.
Beneficial effect of the present invention:
The one, the present invention separates metabolite part fungal diseases of plants pathogenic bacteria is had stronger inhibiting Aspergillus restrictus 28 from the plant garlic, and this provides a kind of microorganism strains for agricultural goes up control fungal diseases of plants pathogenic bacteria;
The 2nd, the invention provides the method for utilizing Aspergillus restrictus 28 fermentations to obtain active metabolite; This meta-bolites can be applicable to the control of fungal diseases of plants, for the exploitation of disinfectant use in agriculture has increased new approach.
Description of drawings
Aspergillus restrictus bacterium among the present invention (Aspergillus restrictus) is named and is Aspergillus restrictus bacterium (Aspergillus restrictus) 28.This bacterial strain is in China Committee for Culture Collection of Microorganisms's common micro-organisms center preservation, preservation date on December 31st, 2010, preservation registration number CGMCC No.4507.
Embodiment
Embodiment 1: (separation of Aspergillus restrictus 28 and screening)
The present invention gathers garlic in April, 2009 in vegetable garden, Zhejiang Hangzhou countryside; Through surface sterilization, endogenetic fungus separation, cultivation, fermentation, determination of activity and to suppressing the steps such as screening of fungal diseases of plants pathogenic bacteria effective strain; Therefrom obtain Aspergillus restrictus bacterium (Aspergillus restrictus) 28, preserve.
Aspergillus restrictus bacterium (Aspergillus restrictus) 28 bacteriostatic activities are measured: the fermented supernatant fluid 1mL of the 0.22 μ m membrane filtration of learning from else's experience is in sterile petri dish; Be cooled to 50 ℃ the rapid mixing of PDA substratum with 9mL; Cucumber rhizoctonia rot bacterium (Rhizoctonia solani) that it is 4mm that the cooling back is put 1 diameter respectively on each substratum plane or Rhizoctonia solani Kuhn (Thanatephorus cucumeris) etc. supply examination bacterium bacterium cake; The bacterium cake is connected to petridish central authorities (the petridish diameter is 9cm); Put 28 ℃ of cultivation 36h in the incubator, handle as contrast, repeat 3 times with sterilized water; Adopt the right-angled intersection method to measure colony diameter, calculate inhibiting rate with formula:
Mensuration result shows: Aspergillus restrictus bacterium (Aspergillus restrictus) 28 meta-bolitess are respectively 85% and 88% to the inhibiting rate of cucumber rhizoctonia rot bacterium and Rhizoctonia solani Kuhn.
Embodiment 2: (preparing methods of Aspergillus restrictus 28 fermentating metabolism products)
Carry out according to the following steps:
(1) activation culture of bacterial classification: will separate Aspergillus restrictus 28 bacterial classifications of preserving and move on the potato glucose PDA solid plate, and under 28 ℃ ± 1 ℃ condition, cultivate, and cover whole petridish to mycelia, using diameter is that the punching of 4mm punch tool obtains the bacterium cake, supplies inoculation usefulness;
(2) fermentation culture: fermentation culture is to contain soybean cake powder 20g among every 1000mL, Semen Maydis powder 15g, Zulkovsky starch 20g, wheat bran 10g, peptone 8g, NH
4NO
33g, (NH
4)
2SO
45g, CaCO
35g, MgSO
41g, the bottled fermentation culture 60mL of every 300mL triangle; Prepare back 121 ℃ the sterilization 20min, to be cooled to 40 ℃ under aseptic condition picking 1 diameter be 4mm bacterium cake, under 28 ℃ ± 1 ℃ condition, 180 rev/mins of shaking speed, fermentation culture 7 days;
(3) fermented product extracts: centrifugal collection fermented liquid after fermentation finishes, adopt the equal-volume ethyl acetate extraction, and get the upper strata, and under vacuum condition, be concentrated into medicinal extract, get acetic acid ethyl ester extract, supply bacteriostatic activity to measure.
Following examples 3,28 meta-bolitess of the Aspergillus restrictus described in 4 are embodiment 2 products made therebies; The percentage composition of said product is mass/volume per-cent.
Embodiment 3: (Aspergillus restrictus 28 meta-bolitess are to the restraining effect of Rhizoctonia solani Kuhn and cucumber rhizoctonia rot bacterium)
(1) accurately take by weighing Aspergillus restrictus 28 meta-bolites 30mg, and add the sterilized water of 10mL volume, ultrasonic oscillation fully dissolves, and finally is configured to the mother liquor that concentration is 3000mg/L;
(2) bacteriostatic activity is measured: get above-mentioned mother liquor and be diluted to the solution that concentration is 1250mg/L, 1000mg/L and 500mg/L respectively; Get each concentration solution 1mL in sterile petri dish; Be cooled to 50 ℃ the rapid mixing of PDA substratum (Aspergillus restrictus 28 meta-bolites final concentrations are 125mg/L, 100mg/L and 50mg/L) with 9mL; Rhizoctonia solani Kuhn or cucumber rhizoctonia rot bacterium bacterium cake that it is 4mm that the cooling back is put 1 diameter respectively on each substratum plane, the bacterium cake is connected to petridish central authorities (the petridish diameter is 9cm), puts 28 ℃ of cultivation 36h in the incubator; Handle as contrast with jingganmycin medicament and sterilized water, repeat 3 times; Adopt the right-angled intersection method to measure colony diameter, calculate inhibiting rate with formula:
Test Example: (product of the present invention and conventional pesticide control effect simultaneous test)
Test-results is seen table 1.Find out that from table 1 it is slightly excellent than the conventional chemical medicament that Aspergillus restrictus 28 meta-bolitess suppress effect, and safety.
Table 1 Aspergillus restrictus 28 meta-bolitess are to the inhibition effect of Rhizoctonia solani Kuhn and cucumber rhizoctonia rot bacterium
"-" expression not test (N.T.).
Claims (1)
1. Aspergillus restrictus bacteria strain; It is characterized in that called after Aspergillus restrictus bacterium (Aspergillus restrictus) 28, depositary institution: China Committee for Culture Collection of Microorganisms common micro-organisms center; Preservation day: on December 31st, 2010, preservation registration number is CGMCC No.4507.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201110088205A CN102191185B (en) | 2011-04-08 | 2011-04-08 | Biological control endophytic fungi - Aspergillus restrictus |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201110088205A CN102191185B (en) | 2011-04-08 | 2011-04-08 | Biological control endophytic fungi - Aspergillus restrictus |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN102191185A CN102191185A (en) | 2011-09-21 |
| CN102191185B true CN102191185B (en) | 2012-09-05 |
Family
ID=44600060
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201110088205A Expired - Fee Related CN102191185B (en) | 2011-04-08 | 2011-04-08 | Biological control endophytic fungi - Aspergillus restrictus |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN102191185B (en) |
Families Citing this family (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN109706083B (en) * | 2018-12-10 | 2023-05-30 | 贵州省中国科学院天然产物化学重点实验室(贵州医科大学天然产物化学重点实验室) | Eucommia ulmoides endophytic fungus with remarkably increased antibacterial activity after metabolic regulation |
| CN109504610B (en) * | 2018-12-12 | 2021-11-19 | 安徽中医药大学 | Endophytic fungus Aspergillus sp.MBL1612 extract and application thereof |
Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1279581A (en) * | 1997-10-23 | 2001-01-10 | 阿克佐诺贝尔英国有限公司 | Method of controlling house dust mites and bedmites |
| US7901927B1 (en) * | 2005-08-09 | 2011-03-08 | The United States Of America As Represented By The Secretary Of Agriculture | Transfer and incorporation of heritable symbiotic fungi into non-host plants |
Family Cites Families (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US8080256B2 (en) * | 2007-11-09 | 2011-12-20 | Regents Of The University Of California | Endophytic fungi from Pteromischum sp. plant, compounds and methods of use |
-
2011
- 2011-04-08 CN CN201110088205A patent/CN102191185B/en not_active Expired - Fee Related
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1279581A (en) * | 1997-10-23 | 2001-01-10 | 阿克佐诺贝尔英国有限公司 | Method of controlling house dust mites and bedmites |
| US7901927B1 (en) * | 2005-08-09 | 2011-03-08 | The United States Of America As Represented By The Secretary Of Agriculture | Transfer and incorporation of heritable symbiotic fungi into non-host plants |
Non-Patent Citations (2)
| Title |
|---|
| 俞晓平.植物内生菌及其代谢产物在生物农药创制中的应用.《浙江农业学报》.2006,第18卷(第5期),289-293. * |
| 申屠旭萍.雷公藤内生真菌的分离即活性菌株的筛选.《浙江农业学报》.2006,第18卷(第5期),308-312. * |
Also Published As
| Publication number | Publication date |
|---|---|
| CN102191185A (en) | 2011-09-21 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN102204570B (en) | Application of alternaria alternate metabolic products in cucumber rhizoctonia solani prevention and control | |
| CN102217656B (en) | Application of alternaria alternata metabolite to controlling Botrytis cirerea | |
| CN102191184B (en) | Biocontrol endophytic fungi-Alternaria alternata | |
| CN101445785B (en) | Aspergillus niger fungus capable of poisoning plant parasitic nematodes, preparation method and application thereof | |
| CN102204571B (en) | Application of Alternaria alternata metabolic product in controlling watermelon fusarium oxysporum | |
| CN106661539A (en) | Homogeneous microorganism extract using useful and functional microorganisms and method for producing same | |
| CN101182468A (en) | Trichoderma epiphyte having nematicidal activity as well as preparation method and uses thereof | |
| CN102204572B (en) | Use of metabolites of nigrospora oryzae 46 in prevention of colletotrichum lindemuthianum | |
| CN101940211B (en) | Application of toyocamycin in preventing and curing cucumber wilt | |
| CN103525748A (en) | Production method of Trichoderma harzianum chlamydospore | |
| CN102191185B (en) | Biological control endophytic fungi - Aspergillus restrictus | |
| CN102210329B (en) | Application of Aspergillus restrictus 28-generation product on preventing and curing cucumber Rhizoctonia solani | |
| CN103087949B (en) | Biocontrol endophytic actinomycetes- streptomyces sioyaensis | |
| CN103222479B (en) | Use of Streptomyces sioyaensis metabolite in prevention and control of Colletotrichum lindemuthianum | |
| CN103222482B (en) | Use of Streptomyces sioyaensis metabolite in prevention and control of Alternaria solani | |
| CN102204569B (en) | Application of Aspergillus restrictus 28 metabolic product in controlling rice thanatephorus cucumeris | |
| CN109112163B (en) | Lactobacillus pentosus fermentation liquor and application thereof in inhibiting phytophthora capsici | |
| CN102210331B (en) | Application of Nigrospora oryzae 46 metabolic product on preventing and treating Rhizoctonia solani | |
| CN102210330B (en) | Application of metabolites of nigrospora oryzae 46 in prevention and treatment of botrytis cinerea | |
| CN105441485A (en) | Method for simultaneously preparing leaf application and root application agricultural biocontrol agents and application of leaf application and root application agricultural biocontrol agents | |
| CN103222483B (en) | Use of Streptomyces sioyaensis metabolite in prevention and control of Botrytis cinerea | |
| CN101798564A (en) | Chlorimuron-ethyl degrading bacterium, soil bioremediation agent based on same, and application thereof | |
| CN103222480B (en) | Use of Streptomyces sioyaensis metabolite in prevention and control of Rhizoctonia solani | |
| CN102191186B (en) | Biocontrol endophytic fungus-Nigrospora oryzae | |
| CN103222481B (en) | Use of Streptomyces sioyaensis metabolite in prevention and control of Thanatephorus cucumeris |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| C14 | Grant of patent or utility model | ||
| GR01 | Patent grant | ||
| CF01 | Termination of patent right due to non-payment of annual fee |
Granted publication date: 20120905 Termination date: 20150408 |
|
| EXPY | Termination of patent right or utility model |