A conventional PCR technique to detect <i>Septoria tritici</i> in wheat seeds

Abstract

A PCR assay was developed for detection of wheat seed naturally contaminated with <i>Septoria tritici</i>. <i>S. tritici</i> specific primers were derived from strict alignment of ITS and α-tubulin sequences of the pathogen. The specificity of four sets of synthesised oligonucleotide pairs (A, B, C and D) were tested using isolates from S. tritici, other selected fungi and wheat seeds. A single DNA fragment was amplified from <i>S. tritici</i> isolates with all primer pairs, whereas no product was generated from other DNA sources. <i>S. tritici</i> was also detected in wheat seed lots collected from plants with variable pycnidial coverage on the upper two leaves. PCR detection of as little as 0.5 pg of <i>S. tritici</i> genomic DNAwas possible. This is the first report on the detection of <i>S. tritici</i> DNA in naturally infested wheat seeds. This PCR based assay is simple, rapid, specific, sensitive and suitable for routine detection of the wheat pathogen in infested wheat seeds.