Authors: Mehrdad Madani Len J Ward Solke H De Boer
Publish Date: 2011/03/06
Volume: 130, Issue: 3, Pages: 271-285
Abstract
The heatshock gene Hsp90 was targeted as a new variable genomic region to supplement other DNAbased tests for identification and discrimination of Globodera pallida G rostochiensis and G tabacum tabacum Populations of the potato cyst nematodes G pallida and G rostochiensis PCN originating from Canada France Belgium and USA together with two populations of G tabacum tabacum from the USA and France were used for the amplification of a fragment of the Hsp90 gene General and specific primers and probes for each species were derived from the consensus and nonconsensus regions of the aligned sequences respectively A triplex conventional PCR assay using a general forward and reverse or three specific reverse primers as well as a realtime PCR using general primers and specific TaqMan probes were developed Melting curve analysis and restriction fragment polymorphisms using high resolution electrophoresis were explored for identifying PCR amplicons that characterized and discriminated the three Globodera species in both pure and mixed samples Results from the different molecular assay strategies confirmed the usefulness of Hsp90 as a new additional gene target and showed that several different test options could be used for discrimination of PCNWe thank Steve Wood Guy Belair and Michael Rott for providing Globodera populations from Newfoundland Quebec and British Columbia in Canada respectively and Xiaohong Wang for providing DNA from Globodera populations in New York and Idaho USA We also thank JA LaMondia for providing cysts of G tabacum tabacum and D Mugniery and Nicole Viaene from INRA France and ILVO Belgium for providing cysts of Globodera spp
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