Aspergillus niger strain having highly-resistant activity to carbendazol and uses thereof

A technology of Aspergillus niger and carbendazim, which is applied in the field of Aspergillus niger fungal strains and related characteristics, and can solve the problems of unstable control effect of biocontrol strains in the field

Inactive Publication Date: 2008-05-21
SHENYANG AGRI UNIV
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

At the same time, it was found that the resistant strain has good compatibility with most chemical agents, and ANUV90 can be used synergistically with most commo

Method used

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  • Aspergillus niger strain having highly-resistant activity to carbendazol and uses thereof
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  • Aspergillus niger strain having highly-resistant activity to carbendazol and uses thereof

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Experimental program
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Effect test

Embodiment 1

[0014] The sensitivity and stability of embodiment 1 Aspergillus niger ANUV90 to carbendazim

[0015] Inoculate Aspergillus niger ANUV90 on drug-containing PDA medium plates containing 50 μg / ml, 100 μg / ml, 500 μg / ml, and 1000 μg / ml of carbendazim active ingredients, and measure the colony size after a certain period of time. The mutant strains that acquired drug resistance after mutagenesis were inoculated on normal PSA medium without carbendazim and cultured for 10 consecutive generations, and the genetic stability of drug resistance of Aspergillus niger after mutagenesis was determined.

[0016] Production of drug-resistant medium: 50% carbendazim wettable powder was first dissolved in 0.1mol / L dilute hydrochloric acid to prepare a drug-containing active ingredient of 3750μg / mL (0.375g drug + 100mL sterile water to obtain 3750μg / mL ) mother solution, gradiently diluted to a concentration of 0.375 μg / mL, 37.5 μg / mL, 375 μg / mL, and 3750 μg / mL, and then 1 mL was added to a volu...

Embodiment 2

[0028] Embodiment 2 Aspergillus niger ANUV90 thalline characteristics

[0029] Observe the hyphal morphology of the strain before and after the mutagenesis under a microscope at 40 times, observe the spore morphology through a scanning electron microscope, and take pictures.

[0030] Aspergillus niger ANUV90 can be seen under the same 40 times optical microscope that the hyphae of the drug-resistant mutant strain are narrower and thinner than the original strain, which is likely to be the result of ultraviolet radiation variation. There is no obvious difference in morphology, only slightly irregular (the left picture of attached drawing 1 shows the mycelial form of AN, the middle picture shows the mycelial form of ANUV90, and the right picture shows the morphology of ANUV90 spore heads under scanning electron microscope).

Embodiment 3

[0031] Compatibility of embodiment 3 Aspergillus niger drug-resistant mutant strain ANUV90 with commonly used chemical agents

[0032] Commonly used fungicides (Krew, chlorothalonil, thiophanate-methyl, mancozeb, metalaxyl, Shuangliu, Dimethocarb), insecticides (dimetholone), herbicides (acetochlor, Gibberellin) and plant growth regulator (gibberellin) were used to make the PSA medium containing drug active ingredients at concentrations of 0.01 μg / mL, 0.1 μg / mL, 1 μg / mL, 10 μg / mL, and 100 μg / mL, respectively. Drug-containing as a control. After the plate is prepared, use a sterile puncher with a diameter of 5 mm to obtain bacterial sheets of Aspergillus niger strains and drug-resistant mutant strains of Aspergillus niger, and inoculate them on the drug-containing PSA medium. Cultivate at a constant temperature at 25°C, record the expansion of the colony after 2 days, and measure the diameter of the colony by the cross method.

[0033] PSA medium: potato 200g, sucrose 20g, ag...

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Abstract

The present invention relates to an Aspergillus niger strain ANUV90 with high stable resistance to carbendazim obtained by mutagenesis, and the resistance of the strain to carbendazim is 1000 times higher than that of the original starting strain AN. Aspergillus niger ANUV90 can still grow normally under the treatment concentration of active ingredient carbendazim up to 1000 μg/mL, and the fermentation broth of this strain also has nematicidal activity. After testing, the strains before and after mutagenesis have no significant difference in colony morphology and other characteristics. At the same time, they have certain biological activity against tomato root-knot nematode. The preliminary research results of the analysis of the molecular mechanism of drug resistance of the mutant strains show that the gene encoding β-tubulin has a site-directed mutation , leading to the generation of drug resistance in biological characteristics of ANUV90. The bacterial strains specifically involved in the present invention have been preserved in the General Microorganism Center of China Microbiological Culture Collection Management Committee on October 30, 2007, with the preservation number CGMCC No.2234, and the classification name: Aspergillus niger. This feature of the strain allows it to be used synergistically with most commonly used pesticides to control a variety of harmful organisms, providing a way to solve the problem of unstable control effects of biocontrol strains in the field, and has a good development and application prospect.

Description

Technical field: [0001] The invention relates to the field of fungi, in particular to an Aspergillus niger fungal strain resistant to high-concentration carbendazim and related characteristics thereof, and belongs to the field of biotechnology. Background technique: [0002] The introduction of antagonistic microorganisms into the soil to prevent and control plant diseases, especially soil-borne diseases, has been carried out for more than 60 years, and has become the consensus of plant disease biocontrol workers, and has achieved certain results. At present, the use of bio-control microorganisms to replace chemical fungicides has become a direction of efforts, but the promotion and application of bio-control strains almost always encounter the same obstacle, that is, some bio-control bacteria are effective against pathogens in indoor experiments and greenhouse experiments. It exhibits extremely high control efficiency, but the biocontrol effect in the field is unstable. In...

Claims

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Application Information

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IPC IPC(8): C12N1/14C12N13/00A01N63/04A01P5/00C12R1/685
Inventor 段玉玺陈立杰王媛媛杨宁朱晓峰
Owner SHENYANG AGRI UNIV
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