Specificity molecule marker of thick wall cell verticil ZK7 strain and its application

A molecular marker and specific technology, applied in the field of molecular biology, can solve the problems of inability to accurately and objectively reflect the colonization, survival and growth and decline of bio-control bacteria, and the sensitivity and accuracy of bio-control fungi are not high

Inactive Publication Date: 2004-07-21
YUNNAN UNIV
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  • Abstract
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  • Claims
  • Application Information

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Problems solved by technology

However, the above methods have certain limitations. The sensitivity and accuracy in the detection process of bio-control fungi are not high, or they can only be used in specific environmental conditions, and cannot accurately and objectively reflect the determination of bio-control fungi in the natural environment. Reproduction, survival and growth and decline rules
[0005] At the same time, since most of the nematode biocontrol products currently on the market are produced by live bacteria, it is very easy for consumers to isolate the production strain from the product when using them.

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  • Specificity molecule marker of thick wall cell verticil ZK7 strain and its application
  • Specificity molecule marker of thick wall cell verticil ZK7 strain and its application
  • Specificity molecule marker of thick wall cell verticil ZK7 strain and its application

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Embodiment Construction

[0022] The following examples are used to further describe the present invention, but the content of the present invention is not limited thereto.

[0023] 1. Acquisition of specific DNA fragments

[0024] By cultivating the pure culture mycelium of ZK7 strain and control strain, extracting their genomic DNA respectively, carrying out PCR amplification with universal primers, and screening primers with DNA polymorphisms for all tested strains. The amplified DNA polymorphisms were compared, and the primer OPL-02 (5'-TGG GCG TCA A-3') amplified a unique DNA fragment of about 1200bp in ZK7 strain, which was named Vc1200 ; Primer OPD-05 (5'-TGA GCG GAC A-3') amplifies a ZK7 strain-specific DNA fragment with a size of about 2000bp, which is named Vc2000.

[0025] 2. Determination of specific DNA fragment sequence

[0026] The specific DNA fragments Vc1200 and Vc2000 on the agarose gel after electrophoresis were excised under the ultraviolet viewer, and the DNA fragments were reco...

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Abstract

A specific molecular marker of verticillium chlamydosporium ZK7 strain and its application are disclosed. The RAPD PCR amplification is used for molecular fingerprint analysis and comparison of said ZK7 strain, different reference strains and the fungus frequently encountered in soil to obtain the RAPD specific DNA fragments Vc 1200 and Vc 2000, which are then recovered, purified, cloned and sequenced. The specific primer SCAR-PCR is designed for PCR amplification. Said Vc 1200 and Vc2000 are marked by digoxin to obtain hybridization probes, which are used for the spot hydridization with the DNA extracted from the soil where the ZK7 strain is applied, so specifically detecting if there is ZK7 strain in soil and its quantity.

Description

Technical field: [0001] The invention relates to a specific molecular marker of Verticillium chlamydospora ZK7 strain and an application thereof, belonging to the technical field of molecular biology. technical background: [0002] Root-knot nematode (Meloidogyne spp.) is one of the important groups of plant parasitic nematodes, and the annual losses to the world's important economic crops are as high as tens of billions of dollars (Barker et al., 1986). One of the main limiting factors of production. It has caused serious harm in major tobacco-growing areas in my country, and it is becoming more and more serious. The affected area in Yunnan Province alone reaches more than 400,000 mu, and the diseased plants cause a 30-50% reduction in tobacco leaf production (Li Tianfei et al., 1994; Yang Ming et al., 1995). Due to the lack of resistant varieties in production, the high multiple cropping index of crops, the limited use of chemical nematicides and the need for sustainable ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68
Inventor 张克勤祝明亮周薇
Owner YUNNAN UNIV
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