Abstract
Stubby root nematodes are of increasing importance to the potato industry in the Pacific Northwest (PNW) of the USA as they transmitTobacco rattle virus (TRY), the causal agent of corky ringspot disease. In the PNW,Paratrichodorus allius is the most prevalent TRV vector nematode species in potato growing areas. A closely related species,P. teres, is also found in parts of the PNW.P. allius andP. teres are morphologically similar, thus creating problems for routine identification. Few studies have utilized molecular techniques to identify species belonging to the Trichodoridae. Species-specific reverse PCR primers for bothParatrichodorus species were designed and located in the ITS-1 rDNA region and used in combination with a universal nematode forward primer located in the 18S rDNA. Primer reliability was confirmed byin vitro screening of differentP. allius populations andin silico analysis ofP. teres on the respective target species. Furthermore, specificity was demonstrated by the absence of cross-reactions with a range of common plant-parasitic nematode species typical of potato growing areas in the PNW. A duplex PCR test was developed that was effective for simultaneous detection of both targetParatrichodorus species.
Resumen
Los nematodos que causan la raíz tocón son de creciente importancia para la industria papera en el Pacífico Noroeste (PNW) de EUA porque transmiten elTobacco rattle virus (TRV), agente causal del anillo corchoso. En el PNW, elParatrichodorus allius es el más importante vector del TRV en las áreas de cultivo de papa. También está presente en ciertas partes del PNWP. teres, una especie relacionada.P. allius yP. teres son morfológicamente similares, por lo que crean problemas para una identificación de rutina. Algunos estudios han utilizado técnicas moleculares para identificar las especies pertenecientes a los Trichodoridae. Los iniciadores de PCR en reversa, específicos para ambas especies deParatrichodorus han sido diseñados y localizados en la región ITS-1 rADN y usados en combinación de un iniciador en la región 18S rADN. La confiabilidad del iniciador fue confirmada en un tamizadoin vitro de diferentes poblaciones deP. allius y en análisisin silico deP. teres, sobre las respectivas especies objetivo. Más aún, la especificidad fue demostrada por ausencia de reacciones cruzadas con una gama de especies de nematodos parásitos de plantas típicas de las áreas de cultivo de papa en el PNW. Ha sido desarrollada una prueba de PCR doble que fue más efectiva para la detección de simultánea de ambas especies deParatrichodorus.
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Riga, E., Karanastasi, E., Marcelo, C. et al. Molecular identification of two stubby root nematode species. Amer J of Potato Res 84, 161–167 (2007). https://doi.org/10.1007/BF02987139
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DOI: https://doi.org/10.1007/BF02987139