A LAMP Assay for Rapid Detection of Stem Rot Fusarium oxysporum on Succulent Plant, Echeveria Perle von Nürnberg

  Objective   A LAMP (loop-mediated isothermal amplification) assay for rapid, visible and sensitive detection of Fusarium oxysporum on the succulent plant, Echeveria Perle von Nürnberg, was developed for early prevention and control of the disease.

  Method  Based on the elongation factor 1α (EF-1α ) sequence of F. oxysporum , LAMP primers were designed. Template DNA from the infected stems were used to optimize the reaction temperature and time. Specificity and sensitivity of the assay were verified on the detection on infected plants from the field.

  Result   The newly developed LAMP assay at 65 °C for 60 min effectively detected F. oxysporum on Echeveria . The assay gave the positive yellowish green result only on F. oxysporum of Echeveria . It had a detection limit of 10 fg·μL⁻¹ and was 100% positive on detecting F. oxysporum from 15 Echeveria samples showing typical stem rot symptoms.

  Conclusion  The established LAMP assay had high specificity and sensitivity with visual and low false positive results. It was considered suitable for rapid detection of F. oxysporum on Echeveria Perle von Nürnberg in the field.