Detection of Leptosphaeria korrae with the polymerase chain reaction and primers from the ribosomal internal transcribed spacers

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Abstract
Leptosphaeria korrae, the causal agent of necrotic ring spot, is a destructive patch disease of Kentucky bluegrass. To develop a rapid molecular test for the detection of this pathogen, an assay based on the polymerase chain reaction (PCR) was developed utilizing the internal transcribed spacer region 1 (ITS 1) of L. korrae ribosomal DNA. DNA sequence comparison showed 94.8% similarity of the ITS 1 region among L. korrae isolates and only 45–50% similarity between L. korrae and other fungal species. Based on ITS 1 sequence differences, a pair of oligonucleotide primers, LK17S and 5.8SC, were selected. With PCR, the primers specifically amplified L. korrae DNA and did not amplify DNA isolated from 15 other fungal species or healthy Kentucky bluegrass. The assay could also specifically detect L. korrae in diseased turfgrass samples. Key words: detection, ITS 1, Leptosphaeria korrae, necrotic ring spot, polymerase chain reaction, ribosomal DNA.

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