Qualitative Real-Time PCR SYBR Green Detection of Petri Disease Fungi

• Main Authors: B.E. Overton, E.L. Stewart, X. Qu, N.G. Wenner, B.J. Christ
• Format: Article
• Language: English
• Published: Firenze University Press 2004-12-01
• Series: Phytopathologia Mediterranea
• Online Access: https://oajournals.fupress.net/index.php/pm/article/view/5095

Real-time PCR provides a fast, reliable, and cost-effective method for detecting the presence or absence of Petri disease fungi in grapevines. The primer pairs, Pmo1f + Pmo2r, and Pac1f + Pac2r, were designed for species and genus-specific amplification of Phaeomoniella chlamydospora and Phaeoacremonium spp. respectively, using realtime PCR with SYBR® Green. The primers were specific and showed no primer-primer dimers until after 35 cycles. Pa. chlamydospora was detected in roots, shoots, and young trunks of drill-inoculated vines. Phaeoacremonium was detected in trunk cross-sections of naturally infected vines from which Phaeoacremonium aleophilum had been isolated. The protocol presented here can be adapted to provide a reliable detection system for research and industry.