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Development of loop-mediated isothermal amplification assays for the detection of seedborne fungal pathogens Fusarium fujikuroi and Magnaporthe oryzae in rice seed

Lookup NU author(s): Dr Sara Franco OrtegaORCiD, Professor Neil Boonham

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Abstract

© 2018 The American Phytopathological Society. Bakanae disease (caused by Fusarium fujikuroi) and rice blast (caused by Magnaporthe oryzae) are two of the most important seedborne pathogens of rice. The detection of both pathogens in rice seed is necessary tomaintain high quality standards and avoid production losses. Currently, blotter tests are used followed by morphological identification of the developing pathogens to provide an incidence of infection in seed lots. Two loop-mediated isothermal amplification assays were developed with primers designed to target the elongation factor 1-α sequence of F. fujikuroi and the calmodulin sequence of M. oryzae. The specificity, sensitivity, selectivity, repeatability, and reproducibility for each assay was assessed in line with the international validation standard published by the European andMediterranean Plant Protection Organization (PM7/98). The results showed a limit of detection of 100 to 999 fg of DNA of F. fujikuroi and 10 to 99 pg of M. oryzae DNA. When combined with a commercial DNA extraction kit, the assays were demonstrated to be effective for use in detection of the pathogens in commercial batches of infected rice seed of different cultivars, giving results equivalent to the blotter method, thus demonstrating the reliability of the method for the surveillance of F. fujikuroi and M. oryzae in seed-testing laboratories.

Publication metadata

Author(s): Ortega SF, Tomlinson J, Hodgetts J, Spadaro D, Gullino ML, Boonham N

Publication type: Article

Publication status: Published

Journal: Plant Disease

Year: 2018

Volume: 102

Issue: 8

Pages: 1549-1558

Print publication date: 01/08/2018

Acceptance date: 02/04/2018

Date deposited: 26/10/2018

ISSN (print): 0191-2917

ISSN (electronic): 1943-7692

Publisher: American Phytopathological Society

URL: https://doi.org/10.1094/PDIS-08-17-1307-RE

DOI: 10.1094/PDIS-08-17-1307-RE

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