DEVELOPMENT OF SPECIFIC PCR PRIMERS FOR THE AMPLIFICATION OF POLYMORPHIC DNA FROM THE OBLIGATE ROOT PATHOGEN PLASMODIOPHORA-BRASSICAE

The DNA sequence of several putative repetitive/high copy number DNA elements cloned from Plasmodiophora brassicae were used to design PCR primers (10-24 mers) for the amplification of polymorphic DNA From P. brassicae. The amplification products obtained with the use of these primers were compared to the products obtained by amplification with arbitrary 10-mer oligonucleotides. The sequence-generated primers produced a small number of polymorphic, highly amplified Fragments while the arbitrary primers produced a large number of fragments which were difficult to distinguish from each other, and from the amplification products obtained from host DNA. The potential use of these primers in studies on the biology and genetic diversity of P. brassicae is discussed. A simple method of P. brassicae DNA preparation for PCR amplification is described. (C) 1995 Academic Press Limited