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Pepper and tomato bacterial Xanthomonas vesicatoria PFGE modular subtyping method

A technology for scab bacteria and molecular typing, which can be used in biochemical equipment and methods, electrophoresis analysis, analysis materials, etc., and can solve problems such as unreported

Inactive Publication Date: 2015-03-25
HUNAN AGRICULTURAL UNIV
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Problems solved by technology

So far, no research has been reported on the typing of pepper and tomato bacterial scab by pulsed field gel electrophoresis

Method used

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  • Pepper and tomato bacterial Xanthomonas vesicatoria PFGE modular subtyping method
  • Pepper and tomato bacterial Xanthomonas vesicatoria PFGE modular subtyping method

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Embodiment Construction

[0055] Using the low frequency restriction endonuclease screened in the present invention Swa Ⅰ and the set pulsed field gel electrophoresis parameters were used for electrophoresis of bacterial scab pathogens from domestic peppers and tomatoes, and the results are shown in figure 1 , all strains showed a total of 15 specific bands, and each strain showed about 3-8 bands, with a size between 180 and 2200 kb. The band information in the electrophoretic spectrum was made into a 0-1 table, with a band marked as 1, and no band marked as 0. Ntsys 2.1 software was used to perform cluster analysis according to the UPGMA algorithm, and four kinds of pepper and tomato bacterial scab Pathogenic bacteria can be effectively distinguished, indicating that the endonuclease and electrophoresis parameters adopted in the present invention can carry out PFGE molecular typing of capsicum and tomato bacterial scab.

[0056] Concrete operation method of the present invention is as follows:

[...

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Abstract

The invention discloses a pepper and tomato bacterial Xanthomonas vesicatoria PFGE modular subtyping method. According to the method, a modular subtyping operating technique is carried out on pepper and tomato bacterial Xanthomonas vesicatoria by adopting a PFGE method combined with DNAs inside cut rubber blocks of special restriction incision enzyme Sml I (SwaI). A pathogenic bacteria suspension is prepared into rubber blocks after protease K treatment, specific bands are obtained through PFGE after enzyme digestion, information of bands is converted for clustering analysis, and bacterial Xanthomonas vesicatoria on pepper and tomato can be effectively distinguished into four species. An incision enzyme used in the method is adopted for the first time, electrophoresis parameters are automatically optimized, and the method can be used for classification and determination and genetic diversity research of pepper and tomato bacterial Xanthomonas vesicatoria.

Description

technical field [0001] The invention relates to molecular typing technology of plant pathogenic bacteria, in particular to a method for typing pepper and tomato bacterial scab (Bacterial Spot caused by Xanthomonads) by pulse field gel electrophoresis (PFGE). Background technique [0002] Capsicum and tomato scab is a bacterial disease that commonly occurs on Solanaceae plants. The pathogen mainly harms the leaves, stems and fruits of peppers and tomatoes, especially on leaves. Leaf disease, water-soaked, yellow-green small spots initially formed, enlarged to become round or irregular, dark brown lesions with raised edges and sunken center, rough and scab-like. In severe cases, the leaves turn yellow, dry, crack, and fall off early. The stems and fruiting stems are diseased, and water-soaked spots are formed at the beginning, and gradually develop into short brown streaks. The lesions are suberized and raised, and the longitudinal fissures are ulcer-like scabs. Fruit disea...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68C12Q1/04
CPCC12Q1/04G01N2550/00
Inventor 刘清波高必达易图永戴良英
Owner HUNAN AGRICULTURAL UNIV
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