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Application of a PCR-Based Cytoplasm Genotyping Method for Phylogenetic Analysis in Potato

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Abstract

We previously developed five polymerase chain reaction-based markers (T, S, SAC, D, and A) to distinguish potato cytoplasms into six types (M, P, A, W, T, and D). As the applicability of this genotyping method for phylogenetic studies had been questioned, we applied this method to species distantly related to cultivars (four accessions of two tomato species, and 176 accessions of 29 Solanum species). The T marker uncovered two insertions, which, along with a unique S marker band, supported independency of series Piurana. The A and D markers generated unique bands to A- and D-type cytoplasms, respectively, but the SAC marker generated a similar banding pattern to the cytoplasms of both cultivated and their distantly related species. Consequently, while the cytoplasm type definition is validated only among cultivated potatoes and their closely related wild species, the developed markers, with the exception of the SAC marker, could provide useful phylogenetic information.

Resumen

Nosotros desarrollamos previamente cinco marcadores con base a la reacción en cadena de la polimerasa (T, S, SAC, D, y A) para distinguir los citoplasmas de la papa en seis tipos (M, P, A, W, T, y D). Considerando que la aplicabilidad de este método de caracterizar genotipos para estudios filogenéticos había sido cuestionada, aplicamos este método para especies poco relacionadas a variedades (cuatro introducciones de dos especies de tomate y 176 de 29 especies de Solanum). El marcador T descubrió dos inserciones, las cuales, junto con una banda única del marcador S, respaldaron independencia de la serie Piurana. Los marcadores A y D generaron bandas únicas a citoplasmas de los tipos A y D, respectivamente, pero el marcador SAC generó un patrón de bandas similar a los citoplasmas tanto de las cultivadas como de las especies con relaciones distantes. Consecuentemente, mientras que la definición del tipo de citoplasma se valida solo entre papas cultivadas y sus especies silvestres con relación cercana, los marcadores desarrollados, con la excepción del SAC, pudieron proporcionar información filogenética útil.

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Acknowledgments

We thank the US Potato Genebank (NRSP-6), Sturgeon Bay, Wisconsin, United States, for providing the Solanum seeds used in this study, Dr. David M. Spooner for kindly checking taxonomic identities with his updated taxonomy, and Ms. Shizuka Souma for technical assistance. This study was supported by Calbee, Inc., Tokyo, Japan, and Calbee Potato, Inc., Obihiro, Japan.

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Authors and Affiliations

  1. Potato Germplasm Enhancement Laboratory, Obihiro University of Agriculture and Veterinary Medicine, Obihiro, Hokkaido, 080-8555, Japan

    Kazuyoshi Hosaka & Rena Sanetomo

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  1. Kazuyoshi Hosaka
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  2. Rena Sanetomo
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Correspondence to Kazuyoshi Hosaka.

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Hosaka, K., Sanetomo, R. Application of a PCR-Based Cytoplasm Genotyping Method for Phylogenetic Analysis in Potato. Am. J. Potato Res. 91, 246–253 (2014). https://doi.org/10.1007/s12230-013-9344-x

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  • DOI: https://doi.org/10.1007/s12230-013-9344-x

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