Open AccessValidation of a Real-Time Polymerase Chain Reaction Assay for the Identification of Meloidogyne arenaria
Author(s) -
Paula Agudelo,
S. A. Lewis,
B. A. Fortnum
Publication year - 2011
Publication title -
plant disease
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.663
H-Index - 108
eISSN - 1943-7692
pISSN - 0191-2917
DOI - 10.1094/pdis-09-10-0668
Subject(s) - biology , meloidogyne arenaria , polymerase chain reaction , real time polymerase chain reaction , terra incognita , esterase , sybr green i , melting curve analysis , nematode , meloidogyne incognita , genetics , ecology , biochemistry , gene , enzyme
Meloidogyne arenaria is an economically important parasite of many crops worldwide. Identification and detection of this species in soil samples is necessary for the design of crop rotation systems, selection of resistant cultivars, and potential use of biological control options. The objective of this study was to develop and validate a real-time polymerase chain reaction (PCR) assay, using species-specific primers and SYBR Green I Dye, for identification of M. arenaria. The specificity of the assay was confirmed by testing for amplification of DNA from other Meloidogyne spp. and from M. arenaria populations of different geographic origins. Field soil samples containing a mixture of M. arenaria and M. incognita were used to compare identification by the real-time PCR assay with identification by esterase phenotype analysis of mature females and by morphometrics of juveniles. The real-time PCR assay provided an accurate and sensitive means for the identification of single juveniles from soil samples.