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Identification of Single Meloidogyne Juveniles by Polymerase Chain Reaction Amplification of Mitochondrial DNA

T. S. Harris, University of Nebraska-Lincoln
L. J. Sandall, University of Nebraska-Lincoln
Thomas O. Powers, University of Nebraska-Lincoln

Document Type Article

Published in Journal of Nematology (1990) 22(4):518-524. © The Society of Nematologists 1990; Used by Permission


Polymerase chain reaction (PCR) was used to amplify a specific 1.8-kb sequence of mitochondrial DNA from single juveniles and eggs from 17 populations of Meloidogyne incognita, M. hapla, M. javanica, and M. arenaria. Approximately 2 μg amplified product were produced per reaction. Restriction digestion of the amplified product with HinfI permitted discrimination of clonal lineages of the four species. Meloidogyne javanica, however, could not be separated from M. hapla by the enzymes used in these experiments. Various amplification conditions and nematode lysis procedures were examined in order to optimize the speed and quality of identifications.