Rapid Identification of Phytophthora ramorum Using Single Strand Conformation Polymorphism of Ribosomal DNA
Ping Kong1 and Chuanxue Hong1
Phytophthora ramorum is an important quarantine pathogen for most parts of the United States and many other countries worldwide. Current approach to identification of this pathogen is primarily based on morphology, which can be difficult and may lead to misidentification due to interspecific overlap and intraspecific variations of diagnostic characters. This study provided an alternative technique to distinguish P. ramorum from its genetically close relatives, and those that are morphologically similar or that have an overlap of host range. The technique consists of three major steps: (i) simplified mycelium boiling DNA extraction, (ii) DNA amplification with ITS primers, and (iii) single strand conformation polymorphism (SSCP) analysis. It takes no more than 6 hr to run through the entire procedure, which can determine identities of 13 to 44 isolates of Phytophthora, depending on the capacity of thermocycler and electrophoresis unit used. Use of the technique described in this work provides rapid and reliable identification of P. ramorum and its close relatives, while requiring minimal training.
1Virginia Polytechnic Institute and State University, Department of Plant Pathology, Physiology, and Weed Science, Hampton Roads Agricultural Research and Extension Center, Virginia Beach, VA 23455
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