Cloning of DNA fragments specific to the pathotype and race of <Emphasis Type="Italic">Verticillium dahliae</Emphasis> |
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Authors: | Toshiyuki Usami Shu Ishigaki Hiroko Takashina Yuko Matsubara Yoshimiki Amemiya |
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Institution: | (1) Faculty of Horticulture, Chiba University, Matsudo Chiba, 271-8510, Japan |
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Abstract: | Japanese isolates of Verticillium dahliae, a causal agent of wilt disease in many plants, are classifiable into pathotypes based on their pathogenicity. Because these
pathotypes are morphologically indistinguishable, establishing a rapid identification method is very important for the control
of this pathogen in Japan. For cloning DNA fragments that are useful for identification and specific detection of V. dahliae pathotypes, we performed random amplified polymorphic DNA (RAPD) analyses using various isolates. One polymerase chain reaction
(PCR) product, E10-U48, was specific to isolates pathogenic to sweet pepper. The other product, B68-TV, was specific to race
1 of isolates pathogenic to tomato. The specificity of these sequences was confirmed by genomic Southern hybridization. Further
analyses revealed that the region peripheral to B68-TV obtained from the genomic DNA library includes the sequence specific
to all isolates pathogenic to tomato (races 1 and 2). Moreover, sequence tagged site (STS) primers designed from B68-TV and
its peripheral region showed race-specific and pathotype-specific amplification in a PCR assay. The probes and primers obtained
in this study are likely to be useful tools for the identification and specific detection of pathotypes and races of V. dahliae.
The nucleotide sequence data reported are available in the DDBJ/EMBL/GenBank databases under accession number AB095266. |
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Keywords: | Verticillium dahliae Pathotype Race RAPD DNA marker PCR |
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