Journal article
Detection and Discrimination of Pratylenchus neglectus and P. thornei in DNA Extracts from Soil
Plant disease, Vol.92(11), pp.1480-1487
11/2008
Handle:
https://hdl.handle.net/2376/106649
PMID: 30764446
Abstract
A species-specific polymerase chain reaction (PCR) method was developed to detect and identify the root-lesion nematodes Pratylenchus neglectus and P. thornei from soil. A primer set was designed from Pratylenchus 28S rRNA gene sequences of the D3 expansion domain. Primer specificity was confirmed with 23 isolates of 15 nematode species and other plant-parasitic and non-plant-parasitic nematodes typically present in the soil communities, and with six fungal species commonly associated with wheat root rot. DNA obtained using a commercially available kit and a method developed in our laboratory gave comparable amplification. PCR conditions were optimized and the two species were differentiated by PCR products of 144 bp for P. neglectus and 288 bp for P. thornei. With this assay, we detected a single juvenile in 1 g of sterile, inoculated soil. Examination of 30 field soil samples revealed that this method was applicable to a range of soils naturally infested with these two pathogens in Oregon. This PCR-based method is rapid, efficient, and reliable, does not require expertise in nematode taxonomy and morphology, and could be used as a rapid diagnostic tool for commercial and research applications for disease forecasting and management.
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Details
- Title
- Detection and Discrimination of Pratylenchus neglectus and P. thornei in DNA Extracts from Soil
- Creators
- Guiping Yan - Oregon State University, Columbia Basin Agricultural Research Center, Pendleton 97801Richard W Smiley - Oregon State University, Columbia Basin Agricultural Research Center, Pendleton 97801Patricia A Okubara - United States Department of Agriculture–Agricultural Research Service (USDA-ARS), Root Disease and Biological Control Research Unit, Pullman, WA 99164Andrea Skantar - USDA-ARS, Nematology Laboratory, Beltsville, MD 20705Sandra A Easley - Oregon State University, Columbia Basin Agricultural Research Center, PendletonJason G Sheedy - Oregon State University, Columbia Basin Agricultural Research Center, PendletonAlison L Thompson - Oregon State University, Columbia Basin Agricultural Research Center, Pendleton
- Publication Details
- Plant disease, Vol.92(11), pp.1480-1487
- Academic Unit
- Plant Pathology, Department of
- Identifiers
- 99900546818801842
- Language
- English
- Resource Type
- Journal article