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Cloning of the mating type locus from Ascochyta lentis (teleomorph: Didymella lentis) and development of a multiplex PCR mating assay for Ascochyta species

Current Genetics volume 50, pages 203–215 (2006)Cite this article

Abstract

The mating type (MAT) locus of the lentil pathogen, Ascochyta lentis, was cloned and characterized using thermal asymmetric interlaced and inverse PCR with primers designed to the HMG-box of Ascochyta rabiei. A multiplex PCR assay for mating type was developed based on MAT idiomorph and flanking sequences. Primers were designed to specifically amplify MAT from several Ascochyta spp. including A. pisi, A. fabae and A. viciae-villosae in addition to A. lentis. Four hundred and fifty and 700 bp fragments were amplified from MAT1-1 and MAT1-2 isolates, respectively, and fragment size correlated perfectly with laboratory crosses using mating type tester strains. MAT-specific PCR allowed rapid scoring of mating type in crude DNA extracts from geographically diverse population samples of A. viciae-villosae from California and Washington State, USA. This co-dominant MAT-specific PCR assay will be a valuable tool for studying the population structure, biology and epidemiology of these fungi.

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Acknowledgments

A portion of this research was conducted during the visit of M. Chérif to the laboratory of T.L. Peever supported by a Fulbright Fellowship and USDA-STEEP for partial funding. The authors would like to thank Tamara Horton for technical assistance. Nucleotide sequence data reported are available in the GenBank database under the accession numbers DQ341314 and DQ341315.

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  1. Walter J. Kaiser

    Present address: 3394 Chickory Way, Boise, ID, 83706, USA

Authors and Affiliations

  1. Laboratoire de Phytopathologie, Institut National Agronomique de Tunisie, 43 Avenue Charles Nicolle 1082, Cité Mahrajéne, Tunis, Tunisia

    Mohamed Chérif

  2. Department of Plant Pathology, Washington State University, Pullman, WA, 99164-6430, USA

    Martin I. Chilvers, Hajime Akamatsu & Tobin L. Peever

  3. USDA-ARS, Washington State University, Pullman, WA, 99164-6430, USA

    Walter J. Kaiser

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  1. Mohamed Chérif
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  2. Martin I. Chilvers
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  3. Hajime Akamatsu
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  4. Tobin L. Peever
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Correspondence to Tobin L. Peever.

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Communicated by U. Kück

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Chérif, M., Chilvers, M.I., Akamatsu, H. et al. Cloning of the mating type locus from Ascochyta lentis (teleomorph: Didymella lentis) and development of a multiplex PCR mating assay for Ascochyta species. Curr Genet 50, 203–215 (2006). https://doi.org/10.1007/s00294-006-0085-y

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  • DOI: https://doi.org/10.1007/s00294-006-0085-y

Keywords

  • Ascochyta lentis
  • High mobility group
  • HMG box
  • Inverse PCR
  • Thermal asymmetric interlaced (TAIL)-PCR
  • Multiplex PCR