NDLI: A loop-mediated isothermal amplification-based method for confirmation of Guignardia citricarpa in citrus black spot lesions

Content Provider	Springer Nature Link
Author	Tomlinson, J. A. Ostoja Starzewska, S. Webb, K. Cole, J. Barnes, A. Dickinson, M. Boonham, N.
Copyright Year	2013
Abstract	Guignardia citricarpa Kiely (anamorph Phyllosticta citricarpa Van der Aa), the causal agent of citrus black spot disease, is subject to phytosanitary restrictions in the EU and USA, such that consignments of citrus are rejected at import if citrus black spot is identified on inspection. Due to the variability of black spot symptoms, positive identification solely on the basis of visual inspection is difficult, especially when lesions lack pycnidia (fruiting bodies of the anamorph Phyllosticta citricarpa). As an aid to visual inspection of symptoms, we have developed a method for detection of G. citricarpa using loop-mediated isothermal amplification (LAMP) which can be used to confirm the presence of G. citricarpa in black spot lesions, including those lacking pycnidia. The LAMP assay can be used to test crude extracts prepared directly from lesions on fruit, and the entire test can be completed in less than 40 min, making it faster than previously described PCR-based methods for detection of G. citricarpa. The method is sufficiently simple to allow deployment of the test in the field, for example in the course of import inspections. Recent years have seen the description of a number of newly recognised species in the genus Phyllosticta that are associated with citrus. As new species emerge, and the taxonomy of the genus is resolved, it will be important to periodically re-evaluate the performance of DNA-based methods for detection of G. citricarpa, including the LAMP assay described here, such that the accuracy of diagnosis can be assured.
Starting Page	217
Ending Page	224
Page Count	8
File Format	PDF
ISSN	09291873
Journal	European Journal of Plant Pathology
Volume Number	136
Issue Number	2
e-ISSN	15738469
Language	English
Publisher	Springer Netherlands
Publisher Date	2013-01-25
Publisher Place	Dordrecht
Access Restriction	One Nation One Subscription (ONOS)
Subject Keyword	Detection Citrus black spot Isothermal amplification Rapid testing Plant Pathology Plant Sciences Ecology Agriculture Life Sciences
Content Type	Text
Resource Type	Article
Subject	Plant Science Horticulture Agronomy and Crop Science

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