Plant Protect. Sci., 2016, 52(3):176-181 | DOI: 10.17221/179/2015-PPS

Identification of Phytophthora species by a high resolution melting analysis: an innovative tool for rapid differentiationOriginal Paper

Antonios Zambounis1, Anastasios Samaras2, Aliki Xanthopoulou3, Maslin Osathanunkul4, Leonardo Schena5, Athanasios Tsaftaris1,3, Panagiotis Madesis3
1 Laboratory of Genetics and Plant Breeding, School of Agriculture, Forestry and Natural Environment, Aristotle University of Thessaloniki, Thessaloniki, Greece
2 Laboratory of Phytopathology, School of Agriculture, Forestry and Natural Environment, Aristotle University of Thessaloniki, Thessaloniki, Greece
3 Institute of Applied Biosciences, CERTH, Thermi, Thessaloniki, Greece
4 Department of Biology, Faculty of Science, Chiang Mai University, Chiang Mai, Thailand
5 Departement of Agricultural Science, Faculty of Agricultural Science, Mediterranea University of Reggio Calabria, Reggio Calabria, Italy

A new molecular method via the high resolution melting (HRM) analysis of the Ypt1 gene non-coding regions was validated for ten Phytophthora species with a broad host range from forest trees to crop species. The melting curve analysis of the amplicons specifically grouped all species into 10 respective unique and distinct HRM curve profiles. The analysis of the normalised HRM melting curves, assigning P. nicotianae as a normalised reference genotype, revealed that the genotype similarities among all the species were adequately low, indicating that Ypt1 marker was sufficient to identify and differentiate the tested species. This HRM method is rapid and reproducible allowing the identification of Phytophthora species and the screening of eventual variants eliminating the separate steps and reducing the risk of contamination.

Keywords: fungal differentiation; fungi of vegetable and tree plants; HRM; plant breeding; Ypt1 gene

Published: September 30, 2016  Show citation

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Zambounis A, Samaras A, Xanthopoulou A, Osathanunkul M, Schena L, Tsaftaris A, Madesis P. Identification of Phytophthora species by a high resolution melting analysis: an innovative tool for rapid differentiation. Plant Protect. Sci. 2016;52(3):176-181. doi: 10.17221/179/2015-PPS.
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