Abstract
Russet Burbank, Russet Norkotah, and Shepody tubers infected with Potato virus Y strains (PVYO, PVYN:O, PVYNTN) were tested from storage at 4°C at the initiation of sprouting and then the same tubers were tested again from storage at 4°C seventy eight days later. Samples were taken from eyes in the stem, middle, and bud (distal end) areas of the tubers. Testing of the samples was done with an enzyme-linked immunosorbent assay (ELISA) and negative samples were retested using reverse transcription-polymerase chain reactions (RT-PCR). The ELISA alone and the ELISA+PCR combined results were evaluated for each of the strains, sample locations, sample time, and cultivar. Results show that ELISA underestimated the actual percentage of tubers with virus. When test results were combined to show a more accurate percentage, PVYN:O was unevenly distributed in some cultivars, but PVYO and PVYNTN were evenly distributed 78 days after initial sprouting. Results show that protocols for PVY post-harvest testing used by state seed certification agencies should be written to specify the amount of time needed before sampling and that specific protocols may be needed based on cultivar in order to accurately detect PVY in tuber samples.
Resumen
De un almacenamiento a 4°C, se probaron tubérculos de las variedades Russet Burbank, Russet Norkotah y Shepody, infectados con el virus Y de la papa variantes PVYO, PVYN:O, PVYNTN, para inicio de la brotación, y después, los mismos tubérculos se probaron otra vez de almacenamiento a 4°C setenta y ocho días mas tarde. Las muestras se tomaron de los ojos de las áreas del tallo, en medio, y ápice (extremo distal) de los tubérculos. Las pruebas se hicieron con un ensayo de inmunoabsorción enzimática (ELISA) y se probaron muestras negativas usando reacción en cadena de la polimerasa de transcripción inversa (RT-PCR). Se evaluaron la ELISA sola y los resultados combinados de ELISA+PCR para cada una de las variantes, localidades de la muestra, tiempo de la muestra y variedad. Los resultados muestran que la ELISA subestimó el porcentaje actual de tubérculos con virus. Cuando se combinaron los resultados de las pruebas para mostrar un porcentaje mas preciso, PVYN:O se distribuyó mas desuniformemente en algunas variedades, pero PVYO y PVYNTN estuvieron distribuidos uniformemente después de 78 días de la brotación inicial. Los resultados muestran que los protocolos para las pruebas de PVY de postcosecha utilizados por las agencias estatales de certificación de semilla deberían estar por escrito para especificar la cantidad de tiempo requerido antes del muestreo y que los protocolos específicos serán necesarios basados en la variedad, a fin de detectar con precisión PVY en muestras de tubérculo.
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Acknowledgements
The authors thank Penny Tubbs, Darren Hall, Melinda Bateman, Melissa Bertram, and Jill Randall for their assistance in conducting this research. This work was partially funded by the USDA/CREES Special Potato Grant Program.
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Whitworth, J.L., Hamm, P.B. & Nolte, P. Distribution of Potato virus Y Strains in Tubers during the Post-Harvest Period. Am. J. Pot Res 89, 136–141 (2012). https://doi.org/10.1007/s12230-012-9235-6
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DOI: https://doi.org/10.1007/s12230-012-9235-6