Abstract
Spraing (brown rings or arcs) of potato caused by Potato mop-top virus (PMTV) occurred in a field of Tokachi, Hokkaido, Japan in 2005. To monitor the expansion of spraing-affected areas, we developed a soil diagnostic method that consisted of a bioassay using tomato as bait plant to trap the vector of PMTV, Spongospora subterranea, the causal agent of powdery scab of potatoes, followed by reverse transcription-polymerase chain reaction-microplate hybridization (RT-PCR-MPH) to detect the virus from roots of bait plants. After incubation of tomato seedlings grown with their roots immersed in a soil suspension at 18°C for 9 days, total RNA extracted from bait roots was analyzed by RT-PCR-MPH using PMTV-specific primers and a digoxigenin (DIG)-labeled probe. Soil diagnosis using the present method in an infested area revealed 137 of 224 fields (61.2%) were infested by PMTV although tubers harvested from only one of these fields had spraing.
Resumen
En Hokkaido, Japon, en 2005, se presentaron en el campo estriados (anillos o arcos cafés) en papa, causados por el virus de trapeador apical (Potato mop-top virus, PMTV). Para monitorear la expansión de las áreas afectadas por el estriado, desarrollamos un método de diagnostico al suelo, consistente en un bioensayo usando el tomate como una planta trampa para atrapar al vector del PMTV, Spongospora subterranea, el agente causal de la roña polvorienta de las papas, seguido por una hibridación en microplaca de la reacción en cadena de la polimerasa de reverso-transcripción (RT-PCR-MPH) para detectar al virus de las raíces de las plantas trampa. Después de la incubación de las plántulas de tomate crecidas con sus raíces inmersas en una suspensión de suelo a 18°C por 9 días, se analizó el RNA total de las raíces-trampa con RT-PCR-MPH, utilizando iniciadores específicos para PMTV y una sonda marcada con digoxigenina (DIG). El diagnóstico del suelo utilizando este método en un área infestada reveló que 137 de 224 campos (61.2%) estaban infestados por PMTV, aunque solo los tubérculos cosechados de uno de estos campos tuvieron el estriado.
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Acknowledgments
This research was grant-aided by Research project for utilizing advanced technologies in agriculture, forestry and fisheries (No. 18099), the Agriculture, Forestry and Fisheries Research Council, the Ministry of Agriculture, Forestry and Fisheries.
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Nakayama, T., Maoka, T., Hataya, T. et al. Diagnosis of Potato Mop-Top Virus in Soil Using Bait Plant Bioassay and RT-PCR-microplate Hybridization. Am. J. Pot Res 87, 218–225 (2010). https://doi.org/10.1007/s12230-010-9128-5
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DOI: https://doi.org/10.1007/s12230-010-9128-5