Sensitive and Direct Detection of Heterodera filipjevi in Soil and Wheat Roots by Species-Specific SCAR-PCR Assays
文献类型： 外文期刊
作者： Peng, Huan 1 ; Qi, Xiaoli 1 ; Peng, Deliang 1 ; Long, Haibo 2 ; He, Xufeng 3 ; Huang, Wenkun 1 ; He, Wenting 1 ;
作者机构： 1.Chinese Acad Agr Sci, Inst Plant Protect, State Key Lab Biol Plant Dis & Insect Pests, Beijing 100193, Peoples R China
2.Chinese Acad Trop Agr Sci, Environm & Plant Protect Inst, Danzhou 571737, Peoples R China
3.Hunan Agr Univ, Coll Biosafety Sci & Technol, Changsha 410128, Hunan, Peoples R China
4.Hunan Agr Univ, Hunan Prov Key Lab Biol & Control Plant Dis & Ins, Changsha 410128, Hunan, Peoples R China
期刊名称：PLANT DISEASE （ 影响因子：4.438； 五年影响因子：4.7 ）
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收录情况： SCI
摘要： 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 ul second-stage juvenile (J2) lysate, 3.9 x 10~3 pi adult female lysate, and 10~3 pi cyst ly-sate. 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. Thisstudy is the first to provide a definitive diagnostic assay for H. filipjevi in wheat roots and .soil.
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