A Genetic Marker Associated with the A1 Mating Type Locus in Phytophthora infestans

  • KIM KWON-JONG (Division of Bio-resources Technology, College of Agriculture and Life Sciences, Kangwon National University) ;
  • EOM SEUNG-HEE (Kangwon Agricultural Technology Administration) ;
  • LEE SANG-PYO (Division of Bio-resources Technology, College of Agriculture and Life Sciences, Kangwon National University) ;
  • JUNG HEE-SUN (Division of Bio-resources Technology, College of Agriculture and Life Sciences, Kangwon National University) ;
  • KAMOUN SOPHIEN (Department of Plant Pathology, Ohio State University) ;
  • LEE YOUN SU (Division of Bio-resources Technology, College of Agriculture and Life Sciences, Kangwon National University)
  • Published : 2005.06.01

Abstract

Sexual reproduction plays an important role in the biology and epidemiology of oomycete plant pathogens such as the heterothallic species Phytophthora infestans. Recent worldwide dispersal of A2 mating type strains of P. infestans resulted in increased virulence, gene transfer, and genetic variation, creating new challenges for disease management. To develop a genetic assay for mating type identification in P. infestans, we used the Amplified Fragment Length Polymorphism (AFLP) technique. The primer combination E+AT/M+CTA detected a fragment specific to A1 mating type (Mat-A1) of P. infestans. This fragment was cloned and sequenced, and a pair of primers (INF-1, INF-2) were designed and used to differentiate P. infestans Mat-A1 from Mat-A2 strains. The Mat A1-specific fragment was detected using Southern blot analysis of PCR products amplified with primers INF-1 and INF-2 from genomic DNA of 14 P. infestans Mat-A1 strains, but not 13 P. infestans Mat-A2 strains or 8 other isolates representing several Phytophthora spp. Southern blot analysis of genomic DNAs of P. infestans isolates revealed a 1.6 kb restriction enzyme (EcoRI, BamHI, AvaI)-fragment only in Mat-A1 strains. The A1 mating type-specific primers amplified a unique band under stringent annealing temperatures of $63^{\circ}C-64^{\circ}C$, suggesting that this PCR assay could be developed into a useful method for mating type determination of P. infestans in field material.

Keywords

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