Short Communication Design and Evaluation of PCR Primers for Denaturing Gradient Gel Electrophoresis Analysis of Plant Parasitic and Fungivorous Nematode Communities

A PCR-DGGE primer pair, Tyl2F-Tyl4R, specific to plant parasitic and fungivorous nematodes was designed based on the 18S rRNA gene. The results of community analysis using the primers showed that they are specific to the order Tylenchida. This primer pair detected species belonging to Tylenchida with high sensitivity and high resolution. The number of detected species of plant parasitic and fungivorous nematodes and their band intensity were much improved compared with PCR-DGGE analysis using the SSU18A-SSU9R primer, which is commonly used for nematode community analysis. It was confirmed that using a group-specific primer was effective for nematode community analysis with PCR-DGGE. Key words: detection sensitivity, fungivorous nematode, order Tylenchida, PCR-DGGE primer, plant parasitic nematode A diverse range of nematodes live in all soil ecosystems. They have important roles in soil function, the cycling of plant nutrients and decomposition of organic matter, and are highly sensitive to various environmental factors such as fertilization and plant cultivation (5, 12, 15, 20). Therefore, the analysis of soil nematode communities has evolved as a bioindicator for the evaluation of soil function and th