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Real time PCR assays to detect and quantify the nematodes Pratylenchus vulnus and Mesocriconema xenoplax

Abstract

Economically damaging populations of lesion nematode and ring nematode are managed in tree crops largely through pre-plant chemical fumigation, the use of which is increasingly restricted due to human health and environmental concerns. Reducing the use of fumigants requires precise knowledge of pest nematodes’ density and distribution, however; extensive sampling is costly due to the time intensive process of nematode counting and identification. In this study, species specific primers were designed and real time PCR (qPCR) assays developed separately for both species of nematodes. The assays successfully detected each species and did not show significant amplification of non-target nematode groups. Both assays related well with microscopic counts of prepared solutions of nematodes, as well as solutions extracted from field samples. Such high-throughput molecular quantification could reduce diagnostic costs, allowing a more accurate picture of nematode populations in the field.

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