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Qualitative real-time PCR SYBR Green detection of Petri disease fungi

Overton, B.E., Stewart, E.L., Qu, X., Wenner, N.G., Christ, B.J. ; In: Phytopathologia Mediterranea 43: 403-410. 2004

ABSTRACT:

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 real-time PCR with SYBR ® Green. The primers were specific and showed no primer-primer dimmers 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.