JOURNAL OF NANJING FORESTRY UNIVERSITY ›› 2015, Vol. 39 ›› Issue (06): 1-6.doi: 10.3969/j.issn.1000-2006.2015.06.001

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Rapid diagnostic methods for Phytophthora tentaculata using LAMP

DAI Tingting1,2, KANG Zhenye1,2, WU Xiaoqin1,2   

  1. 1. Co-Innovation Center for the Sustainable Forestry in Southern China, Nanjing 210037, China;
    2. Jiangsu Key Laboratory for Prevention and Control of Invasive Species, College of Forestry, Nanjing Forestry University, Nanjing 210037,China
  • Online:2015-11-30 Published:2015-11-30

Abstract: In order to identify the disease caused by Phytophthora tentaculata, and keep the spread of the disease under control and reduce the economic losses quickly and accurately, loop-mediated isothermal amplification(LAMP)assay targeting the PtYpt1 element was used to detect Phytophthora tentaculata. The PtYpt1-LAMP assay amplified the target element in less than 80 min at 64 ℃ efficiently and was evaluated specificitily and sensitivitily. The specificity was evaluated against Phytophthora tentaculata, Phytophthora spp., Pythium spp., and true fungi isolates. The P. tentaculata DNA products were visualised as a ladder-like banding pattern by 2% gel electrophoresis. A positive colour(sky blue)was only observed in the presence of P. tentaculata by addition of hydroxynaphthol blue prior to amplification, whereas none of other isolates showed a colour change. The detection limit of the PtYpt1-specific LAMP assay for P. tentaculata was 1 ng/μL of genomic DNA per reaction.

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