New polymerase chain reaction (PCR) primers were developed for the identification of the EU quarantine pest Monilinia fructicola. These allowed nine M. ffucticola isolates to be distinguished from other fungi, including six isolates of M. laxa and six isolates of M. fructigena (which also cause brown rot of stone and pome fruit). Three M. fnrcticola isolates from Japan and one from Australia did not react with a primer set previously published for M. fructicola. M. fnrcticola isolates could be subdivided into three groups based on the size of their nuclear rDNA small subunit. The subunits from the four non-reactive isolates were either smaller or larger than the reactive group. Further primers were developed which were specific for either M. laxa or M. fructigena. Another new primer identified both M. laxa and M. fructigena, and yet another M. laxa and M. fructicola. When used in combination, these primers specific for two species correctly identified unknown isolates of all three Monilinia species. The new primers designed in this study have been used to identify, rapidly and correctly, pustules taken directly from infected plum fruits, thus demonstrating their diagnostic potential. and 2 regions of the nuclear rRNA gene repeat were therefore produced ( Table 1). The effectiveness of these primers and the occurrence of group-I introns in isolates of M. fructicola is discussed.
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