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- Title
Species-specific DNA markers for identification of two root-knot nematodes of coffee: Meloidogyne arabicida and M. izalcoensis.
- Authors
Correa, Valdir; Santos, Marcilene; Almeida, Maria; Peixoto, José; Castagnone-Sereno, Philippe; Carneiro, Regina
- Abstract
Seven root-knot nematodes (RKN), including Meloidogyne exigua, M. incognita, M. paranaensis, M. enterolobii, M. arabicida, M. izalcoensis and M. arenaria are major pathogens of coffee crop in the Americas. Species-specific primers for their identification have been developed for five of them and constitute a fast and reliable method of identification. Here we report a PCR-based assay for specific detection of M. arabicida and M. izalcoensis. Random Amplified Polymorphic DNA fragments specific for these two species were converted into sequence characterized amplified region (SCAR) markers. PCR amplification using the SCAR primers produced a specific fragment of 300 bp and 670 bp for M. arabicida and M. izalcoensis, respectively, which were absent in other coffee-associated Meloidogyne spp. tested. SCAR primers also allowed successful amplification of DNA from single second-stage juveniles (J2), males and females. In addition, these primers were able to unambiguously detect the target species in nematode suspensions extracted from soil and roots samples, in different isolates of the same species or when used in multiplex PCR reactions containing mixtures of species. These results demonstrated the effectiveness of these SCAR markers and their multiplex use with those previously developed for M. exigua, M. incognita, M. paranaensis, M. enterolobii and M. arenaria constitute an essential detection tool . This diagnostic kit will contribute for specific J2 identification of the major RKN infecting coffee from field samples in the Americas.
- Subjects
GENETIC markers in plants; ROOT-knot nematodes; COFFEE diseases &; pests; SOUTHERN root-knot nematode; POLYMERASE chain reaction; NEMATODE identification
- Publication
European Journal of Plant Pathology, 2013, Vol 137, Issue 2, p305
- ISSN
0929-1873
- Publication type
Article
- DOI
10.1007/s10658-013-0242-3