Abstract
Consistent and effective methods for early discrimination of pathogen resistance, and selection of times for tissue sampling, are important for experiments using global gene expression and metabolomics. Assays for resistance to the vascular pathogen Verticillium dahliae (Vd), the causal agent of Verticillium wilt (VW), are particularly difficult because escapes are common in field assays. Seedling dip assays offer a potential solution, but homogeneous populations are not typically available. As an alternative strategy, we have developed a protocol for studying spatiotemporal infection dynamics of Vd using potato stem cuttings. The protocol was validated using genotypes varying in resistance/susceptibility to Vd. Although there were no visual symptoms in the plants, stem sections were infested with Vd as early as 7 dpi. Symptoms were first observed in the most susceptible genotype at 10 dpi and became apparent on all test subjects at 14 dpi. The protocol has potential applications in resistance breeding and ‘omics’ studies where populations derived from true seeds are not available.
Resumen
Los métodos consistentes y efectivos para la discriminación temprana de resistencia a patógenos, y la selección de los tiempos para el muestreo de tejidos, son importantes para los experimentos usando expresiones globales de genes y metabolómica. Los ensayos para resistencia al patógeno vascular Verticillium dahliae (Vd), el agente causal del marchitamiento por Verticillium (VW), son particularmente difíciles porque son comunes los escapes en ensayos de campo. Los ensayos de inmersión de plántulas ofrecen una solución potencial, pero poblaciones homogéneas no están disponibles típicamente. Como una estrategia alternativa, hemos desarrollado un protocolo para estudiar la dinámica de infección espaciotemporal de Vd utilizando esquejes de tallo de papa. Se validó el protocolo usando genotipos con variación en resistencia/susceptibilidad al Vd. Aun cuando no había síntomas visibles en las plantas, se infestaron secciones de tallo con Vd tan pronto como en siete dpi (días posteriores a la inoculación). Primero se observaron los síntomas en el genotipo más susceptible a 10 dpi, y se volvió aparente en todos los sujetos a la prueba a los 14 dpi. El protocolo tiene aplicaciones potenciales en el mejoramiento para resistencia y en estudios “omicos” en donde no hay disponibles poblaciones derivadas de semilla verdadera.
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Acknowledgements
Funding for the project, including salary for A.K., was provided by a USDA/ARS-State Cooperative research grant.
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Kumar, A., Jansky, S. & Halterman, D. Potato Stem Cuttings to Study Verticillium dahliae Infection for Resistance Breeding and ‘omics’ Studies. Am. J. Potato Res. 94, 270–274 (2017). https://doi.org/10.1007/s12230-016-9563-z
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DOI: https://doi.org/10.1007/s12230-016-9563-z