LAMP assay to detect Elsinoë necatrix; an important Eucalyptus shoot and leaf pathogen

    Affiliations
    Authors and Affiliations
    • Ms. Alishia van Heerden, University of Pretoria Forestry and Agricultural Biotechnology Institute, 98823, Department of Biochemistry, Genetics and Microbiology, Pretoria, South Africa; [email protected]
    • Dr. Nam Pham, University of Pretoria Forestry and Agricultural Biotechnology Institute, 98823, Department of Plant and Soil Sciences, Pretoria, South Africa; [email protected]
    • Dr. Brenda D. Wingfield, University of Pretoria Forestry and Agricultural Biotechnology Institute, 98823, Department of Biochemistry, Genetics and Microbiology, Pretoria, South Africa, +27829083380; [email protected]
    • Prof. Mike Wingfield, University of Pretoria Forestry and Agricultural Biotechnology Institute, 98823, Department of Biochemistry, Genetics and Microbiology, Pretoria, South Africa, +27829083380; [email protected]
    • Dr. Jupiter Muro Abad, Plant Health Program, Research and Development, Asia Pacific Resources International Holdings Ltd. (APRIL), Pangkalan Kerinci, Riau, Indonesia; [email protected]
    • Dr. Alvaro Duran, Plant Health Program, Research and Development, Asia Pacific Resources International Holdings Ltd. (APRIL), Pangkalan Kerinci, Riau, Indonesia; [email protected]
    • Dr. Markus Wilken, University of Pretoria Forestry and Agricultural Biotechnology Institute, 98823, Lunnon Road, Pretoria, South Africa, 0028; [email protected]

      Published Online:https://doi.org/10.1094/PDIS-01-24-0086-RE

      Eucalyptus scab and shoot malformation caused by Elsinoë necatrix is an emerging disease and a serious threat to the global commercial forestry industry. The disease was first discovered in North Sumatra, Indonesia and now requires a simple and effective method for early pathogen detection. In this study, a rapid and sensitive Loop-mediated isothermal amplification (LAMP) assay was developed for E. necatrix. A unique region in a secondary metabolite gene cluster was used as target for the assay. To test robustness of the assay, LAMP amplification was verified in 15 strains of E. necatrix. A specificity test against 23 closely related Elsinoë species and three fungal species commonly isolated on Eucalyptus showed that the LAMP assay exclusively identified E. necatrix isolates. The assay had a high level of sensitivity, able to detect 0.01 ng (approximately 400 target copies) of pure E. necatrix DNA. Furthermore, using a simple DNA extraction method, it was possible to use this assay to detect E. necatrix in infected Eucalyptus leaves.