Development of a quantitative PCR assay for the detection of Piggotia coryli, the causal agent of hazelnut anthracnose

Abstract

A novel real-time PCR assay was developed for the detection of Piggotia coryli, the causal agent of hazelnut anthracnose. A molecular tool for sensitive detection and quantification of P. coryli in symptomatic and asymptomatic host tissues was required to better understand P. coryli biology and epidemiology. Species-specific primers for P. coryli were designed based on the sequence of the β-tubulin gene. The amplification efficiency of pure target DNA was 93.2%, wet lab-tested limit of detection (LOD) was fixed at 8 pg/PCR reaction with maximum values of repeatability and reproducibility. In samples collected from infected orchards, the assay consistently revealed the presence of the pathogen in all symptomatic specimens, as well as in some asymptomatic tissues. This molecular tool will be of substantial aid in detecting and quantifying P. coryli, even in latent state and in different hazelnut tissues.