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Characterization and distribution of two subtypes of Verticillium longisporum isolated from cabbage fields in Japan

  • Fungal Diseases
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Abstract

Verticillium longisporum is an allodiploid hybrid fungus that consists of at least three groups, A1/D1, A1/D2, and A1/D3. PCR-based analysis of 18S rDNA, 5.8S rDNA-ITS region, mtSSU-rDNA, cytochrome b gene, and mating type gene, as well as RAPD analysis, revealed that Verticillium wilt of cabbage in Gunma Prefecture in Japan is caused by two groups of V. longisporum—A1/D1 and A1/D3—and by V. dahliae. A1/D1 and A1/D3 lineage strains were equally distributed in cabbage fields, whereas V. dahliae strains were most frequently isolated. The proportion of the three types of Verticillium had not changed in 1998 and 2008. Although both V. longisporum strains produced longer conidia than the V. dahliae strains, the A1/D3-type strains produced larger conidia than A1/D1-type strains. In addition, the A1/D3-type strains formed microsclerotia that were distinguishable from those of A1/D1-type of V. longisporum and V. dahliae. The pathogenicity of the A1/D3-type strains on cabbage was similar to that of the A1/D1-type strains. These results will contribute to understanding of genotypic diversity, distribution, and pathogenicity of Verticillium species pathogenic on cabbage.

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Acknowledgments

This work was supported in part by a grant of Strategic Research Foundation Grant-aided Project for Private Universities from the Ministry of Education, Culture, Sports, Science, and Technology of Japan.

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Author notes

  1. Shinpei Banno

    Present address: Japan Plant Protection Association, Kita, Tokyo, 114-0015, Japan

  2. Toshimasa Shiraishi

    Present address: Graduate School of Agriculture and Life Sciences, The University of Tokyo, 1-1-1 Yayoi, Bunkyo-Ku, Tokyo, 113-8657, Japan

Authors and Affiliations

  1. Faculty of Life Sciences, Toyo University, 1-1-1 Izumino, Itakura, Ora, Gunma, 374-0193, Japan

    Shinpei Banno, Hidenari Saito & Makoto Fujimura

  2. Gunma Agricultural Technology Center, 493 Nishi-Obokata, Isesaki, Gunma, 379-2224, Japan

    Kentaro Ikeda, Hiroshi Sakai & Toshimasa Shiraishi

  3. Gunma Agricultural Promotion and Guidance Office, 493 Nishi-Obokata, Isesaki, Gunma, 379-2224, Japan

    Toshihiko Urushibara

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  1. Shinpei Banno
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Corresponding author

Correspondence to Makoto Fujimura.

Electronic supplementary material

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10327_2014_568_MOESM1_ESM.pptx

Supplementary material 1 (PPTX 348 kb). Fig. S1. Polymerase chain reaction (PCR) assay to detect 18S rDNA and 5.8S rDNA in Verticillium strains obtained from cabbage. (a) Gene structure of nuclear rDNA gene of V. dahliae and V. longisporum. (b) PCR amplification of 18S rDNA gene region. M: 1-kb DNA ladder marker (Fermentas Life Sciences, Burlington, ON, Canada) (c) PCR amplification of 5.8S rDNA-ITS region. M: 100-bp ladder marker (New England Biolabs, Ipswich, MA, USA) Genomic DNA isolated from the eight V. longisporum strains (CA1, CA4, CA9, CA10, CA12, CA34, CA54, and CA58), two V. dahliae strains (CA39 and CA43), V. longisporum strain (84013), and V. albo-atrum strain (Vaa-HP) was used as template DNA. Fig. S2. Polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) assays for mitochondrial small-subunit rDNA (mtSSU-rDNA) and cytochrome b genotyping. PCR-RFLP assay of (a) mtSSU-rDNA and (b) cytochrome b. The mtSSU-rDNA and cytochrome b gene region were amplified using primer sets NMS1 plus NMS2 and cobF1 plus cobR1, respectively. PCR products were digested with restriction enzymes (a) MseI and (b) DdeI, respectively. Genomic DNA was purified from the eight Verticillium longisporum strains (CA1, CA4, CA9, CA10, CA12, CA34, CA54, and CA58), two V. dahliae strains (CA39 and CA43), V. longisporum strain (84013), and V. albo-atrum strain (Vaa-HP) and used as template DNAs. M: 100-bp ladder marker (New England Biolabs, Ipswich, MA, USA). Fig. S3. Polymerase chain reaction (PCR) assays to detect the mating type gene in Verticillium strains obtained from cabbage. PCR amplification of (a) MAT1-1-1 and (b) MAT1-2-1. PCR products were analyzed using a MultiNA microchip electrophoresis system (Shimadzu, Kyoto, Japan). Genomic DNA isolated from the eight V. longisporum strains (CA1, CA4, CA9, CA10, CA12, CA34, CA54, and CA58), three V. dahliae strains (CA26, CA39, and CA43), V. longisporum strain (84013), V. dahliae strain (84023), and V. albo-atrum strain (Vaa-HP) was used as template DNA

Supplementary material 2 (DOC 75 kb)

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Banno, S., Ikeda, K., Saito, H. et al. Characterization and distribution of two subtypes of Verticillium longisporum isolated from cabbage fields in Japan. J Gen Plant Pathol 81, 118–126 (2015). https://doi.org/10.1007/s10327-014-0568-5

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  • DOI: https://doi.org/10.1007/s10327-014-0568-5

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