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Development of PCR Assays for the Identification of Species and Pathotypes of Elsinoë Causing Scab on Citrus

July 2007 , Volume 91 , Number  7
Pages  865 - 870

J. W. Hyun , Subtropical Environment Division, National Institute of Subtropical Agriculture, R.D.A. Jeju, 699-803, Korea ; N. A. Peres , University of Florida, Gulf Coast Research and Education Center, Wimauma 33598 ; S.-Y. Yi , National Instrumentation Center for Environmental Management (NICEM), Seoul National University, Seoul, 151-921, Korea ; L. W. Timmer , University of Florida, Citrus Research and Education Center, Lake Alfred 33850 ; K. S. Kim , Citrus Research Division, National Institute of Subtropical Agriculture, R.D.A. Jeju, Korea, 699-803 ; and H.-M. Kwon and H.-C. Lim , Subtropical Environment Division, National Institute of Subtropical Agriculture, Korea



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Accepted for publication 21 February 2007.
ABSTRACT

Two scab pathogens of citrus, Elsinoë fawcettii and E. australis, cause citrus scab and sweet orange scab, respectively, and pathotypes of each species have been described. The two species cannot be readily distinguished by morphological or cultural characteristics and can be distinguished only by host range and the sequence of the internal transcribed spacer (ITS) region. In this study, random amplified polymorphic DNA (RAPD) assays clearly distinguished E. fawcettii and E. australis, and the sweet orange and natsudaidai pathotypes within E. australis also could be differentiated. We developed specific primer sets, Efaw-1 for E. fawcettii; Eaut-1, Eaut-2, Eaut-3, and Eaut-4 for E. australis; and EaNat-1 and EaNat-2 for the natsudaidai pathotype within E. australis using RAPD products unique to each species or pathotype. Other primer sets, Efaw-2 and Eaut-5, which were specific for E. fawcettii and E. australis, respectively, were designed from previously determined ITS sequences. The Efaw-1 and Efaw-2 primer sets successfully identified E. fawcettii isolates from Korea, Australia, and the United States (Florida) and the Eaut-1 to Eaut-5 primer sets identified both the sweet orange pathotype isolates of E. australis from Argentina and the natsudaidai pathotype isolates from Korea. The EaNat-1 and EaNat-2 primer sets were specific for isolates of the natsudaidai pathotype. The Efaw-1 and Efaw-2 primer sets successfully detected E. fawcettii from lesions on diseased leaves and fruit from Korea and primer pairs Eaut-1, Eaut-2, Eaut-3, Eaut-4, and Eaut-5 detected E. australis from lesions on sweet orange fruit from Brazil.



© 2007 The American Phytopathological Society