Development of a real-time fluorescence loop-mediated isothermal amplification assay for rapid and quantitative detection of Fusarium oxysporum f. sp. niveum in soil

Authors

  • Jun Peng,

    1. Key Laboratory of the Ministry of Agriculture for Integrated Pest Management on Tropical Crops, Institute of Environmental and Plant Protection, Chinese Academy of Tropical Agricultural Sciences, Haikou, Hainan, China
    Search for more papers by this author
  • Yuanfeng Zhan,

    1. Tropical Crops Genetic Resources Institute, Chinese Academy of Tropical Agricultural Sciences, Danzhou, Hainan, China
    Search for more papers by this author
  • Fanyun Zeng,

    1. Key Laboratory of the Ministry of Agriculture for Integrated Pest Management on Tropical Crops, Institute of Environmental and Plant Protection, Chinese Academy of Tropical Agricultural Sciences, Haikou, Hainan, China
    Search for more papers by this author
  • Haibo Long,

    1. Key Laboratory of the Ministry of Agriculture for Integrated Pest Management on Tropical Crops, Institute of Environmental and Plant Protection, Chinese Academy of Tropical Agricultural Sciences, Haikou, Hainan, China
    Search for more papers by this author
  • Yuelin Pei,

    1. Key Laboratory of the Ministry of Agriculture for Integrated Pest Management on Tropical Crops, Institute of Environmental and Plant Protection, Chinese Academy of Tropical Agricultural Sciences, Haikou, Hainan, China
    Search for more papers by this author
  • Jianrong Guo

    Corresponding author
    1. Key Laboratory of the Ministry of Agriculture for Integrated Pest Management on Tropical Crops, Institute of Environmental and Plant Protection, Chinese Academy of Tropical Agricultural Sciences, Haikou, Hainan, China
    • Correspondence: Jianrong Guo, Key Laboratory of the Ministry of Agriculture for Integrated Pest Management on Tropical Crops, Institute of Environmental and Plant Protection, Chinese Academy of Tropical Agricultural Sciences, Haikou 571101, Hainan, China. Tel.: +86 898 66969305; fax: +86 898 66969211; e-mail: guojianrong@hotmail.com

    Search for more papers by this author

Abstract

Fusarium wilt caused by Fusarium oxysporum f. sp. niveum (Fon) is one of the major limiting factors for watermelon production worldwide. Rapid and accurate detection of the causal pathogen is the cornerstone of integrated disease management. In this paper, a real-time fluorescence loop-mediated isothermal amplification (RealAmp) assay was developed for the rapid and quantitative detection of Fon in soil. Positive products were amplified only from Fon isolates and not from any other species or formae speciales of F. oxysporum tested, showing a high specificity of the primer sets. The detection limit of the RealAmp assay was 1.2 pg μL−1 genomic DNA or 103 spores g−1 of artificially inoculated soil, whereas real-time PCR could detect as low as 12 fg μL−1 or 102 spores g−1. The RealAmp assay was further applied to detect eight artificially inoculated and 85 field soil samples. No significant differences were found between the results tested by the RealAmp and real-time PCR assays. The RealAmp assay is a simple, rapid and effective technique for the quantitative detection and monitoring of Fon in soil under natural conditions.

Ancillary