Current Genetics

, Volume 50, Issue 3, pp 203–215 | Cite as

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

  • Mohamed Chérif
  • Martin I. Chilvers
  • Hajime Akamatsu
  • Tobin L. Peever
  • Walter J. Kaiser
Research 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.

Keywords

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

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Copyright information

© Springer-Verlag 2006

Authors and Affiliations

  • Mohamed Chérif
    • 1
  • Martin I. Chilvers
    • 2
  • Hajime Akamatsu
    • 2
  • Tobin L. Peever
    • 2
  • Walter J. Kaiser
    • 3
    • 4
  1. 1.Laboratoire de PhytopathologieInstitut National Agronomique de TunisieTunisTunisia
  2. 2.Department of Plant PathologyWashington State UniversityPullmanUSA
  3. 3.USDA-ARSWashington State UniversityPullmanUSA
  4. 4.3394 Chickory WayBoiseUSA

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