Development of specific PCR primers for diagnosis and quantitative detection of the fungal maize pathogen Kabatiella zeae

The fungus Kabatiella zeae which causes the eyespot disease in maize has become an important leaf disease in the northern regions of Europe. An early and accurate diagnosis is necessary for determining the first occurrence in the field. The visual assessment of this pathogen is often difficult because different spots, which may be parasitic, physiological or genetic complicate the exact diagnosis. The specific identification of the fungus is possible by PCR. Unfortunately, no specific DNA sequences are available in the databases. Hence, shot-gun cloning was done resulting in K. zeae-specific sequences for primer design. Cross testing with maize DNA and other pathogens, including Setosphaeria turcica, Mycosphaerella zeae-maydis and various Fusarium species, revealed that the primers were absolutely specific for K. zeae and can be used for qualitative and quantitative PCR assays.