Xiaoli Qi, and
Deliang Peng, State Key Laboratory for Biology of Plant Diseases and Insect Pests, Institute of Plant Protection, Chinese Academy of Agricultural Sciences, Beijing 100193, China;
Haibo Long, Key Laboratory of Pests Comprehensive Governance for Tropical crops, Ministry of Agriculture, P. R. China, Environment and Plant Protection Institute, Chinese Academy of Tropical Agricultural Science, Danzhou 571737, China;
Xufeng He, College of Biosafety Science & Technology, Hunan Agricultural University and Hunan Provincial Key Laboratory for Biology & Control of Plant Diseases and Insect Pests, Hunan Agricultural University, Changsha 410128, China; and
Wenkun Huang and
Wenting He, State Key Laboratory for Biology of Plant Diseases and Insect Pests, Institute of Plant Protection, Chinese Academy of Agricultural Sciences, Beijing 100193, China
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Accepted for publication 9 April 2013.
Cereal cyst nematodes are the most important plant-parasitic nematodes on cereal crops in wheat producing areas of the world. Heterodera filipjevi was first reported in China in 2010. In this study, species-specific sequence characterized amplified region–polymerase chain reaction (SCAR-PCR) assays for detection and identification of H. filipjevi from infected wheat roots and soil were developed. The species-specific primers were designed according to the randomly amplified polymorphic DNA (RAPD) markers amplified with random primer OPK16. A 646-bp specific fragment of sequence was generated, which characterized amplified regions in H. filipjevi. The detection limitation of the PCR assay was as low as 0.125 μl second-stage juvenile (J2) lysate, 3.9 × 10–3 μl adult female lysate, and 10–3 μl cyst lysate. The method was able to detect the various stages (J2, J3, J4, and female) of H. filipjevi, and a single of nematode in 0.5 g of soil. H. filipjevi was detected by the method in two of six field samples, and one of those samples contained a mixed population of H. filipjevi and H. avenae. This study is the first to provide a definitive diagnostic assay for H. filipjevi in wheat roots and soil.
© 2013 The American Phytopathological Society