Identification of the chromosome region responsible for pathogenicity of Verticillium dahliae on tomato using genetic re
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Identification of the chromosome region responsible for pathogenicity of Verticillium dahliae on tomato using genetic recombination through protoplast fusion Toshiyuki Usami1 · Naoko Iida1 · Keisuke Nakao1 · Ayame Hamano1 · Nana Suzuki1 · Yusuke Ohmura1 · Yoshimi Komiya1 Received: 25 October 2019 / Accepted: 16 April 2020 © The Phytopathological Society of Japan and Springer Japan KK, part of Springer Nature 2020
Abstract The host range of Verticillium dahliae, causal agent of verticillium wilt in various dicot plants, differs among strains, but the mechanism responsible for the host-specific pathogenicity of the strains remains unclear. In this study, protoplast fusion of a tomato-pathogenic and a nonpathogenic strain of V. dahliae was used for genetic recombination of the isolates to localize the fungal genomic region involved in the pathogenicity on tomato. Twenty fusion strains resistant to two antibiotics, hygromycin B and geneticin, were obtained by protoplast fusion between two parental strains resistant to one of these antibiotics. Genomic Southern hybridization probed with telomere sequences revealed that these fusion strains were haploid and inherited chromosomes from both parental strains. Eight fusion strains were pathogenic on tomato. In PCR analysis of the fusion strains using DNA markers specific to a parental strain TV103, two DNA markers (T12 and VDA787) were amplified only in strains pathogenic on tomato. The genomic region mapped around these two DNA markers for a parental strain pathogenic on tomato was similar to that on the map for chromosome 3 of a reference strain (JR2). The analysis of 35 fusion strains with additional DNA markers revealed that one of the markers was completely accorded with pathogenicity of the strains on tomato. Therefore, the genomic region around this DNA marker is possibly involved in pathogenicity of V. dahliae on tomato. Keywords Verticillium wilt · Solanum lycopersicum · Parasexual recombination · Protoplast fusion
Introduction The soilborne plant pathogenic fungus Verticillium dahliae causes vascular wilt disease on many species of dicotyledenous plants. A single strain may typically be capable of invading multiple plant species, with the host range differing among strains (Pegg and Brady 2002). V. dahliae is known to be an asexual fungus (Usami et al. 2009b). However, genetic recombination via a parasexual cycle has been reported (O’Garro and Clarkson 1988, 1992; Typas Electronic supplementary material The online version of this article (https://doi.org/10.1007/s10327-020-00955-x) contains supplementary material, which is available to authorized users. * Toshiyuki Usami [email protected]‑u.jp 1
Graduate School of Horticulture, Chiba University, 648 Matsudo, Matsudo‑city, Chiba 271‑8510, Japan
and Heale 1978). Moreover, the host range of V. dahliae (Usami and Amemiya 2005) and other Verticillium species (McGeary and Hastie 1982) can vary following genetic recombination between strains, indicating that a gene(s) in the genome of tho
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