Applicability of propidium monoazide (PMA) for discrimination between living and dead phytoplankton cells

Abstract

Propidium monoazide (PMA) is a highly selective dye that penetrates only membrane-compromised, dead microbial cells and inhibits both DNA extraction and amplification. PMA has been widely used for discrimination between living and dead microbial cells; however, the application of PMA in phytoplankton studies has been limited. In this study, we attempted to evaluate its applicability for the discrimination of viable phytoplankton. We tested PMA on seven phytoplankton species, Microcystis aeruginosa, Anabaena sp., Aphanizomenon sp., Synechocystis sp., Cryptomonas ovata, Scenedesmus obliquus, and Nitzschia apiculata as representatives of the major phytoplankton taxa Cyanobacteria (first four species), Chloro-phyta, Cryptophyta, and Bacillariophyta, respectively. Our results showed that application of PMA to phytoplankton living in freshwater has the potential to distinguish viable from dead cells as in microbial studies. Particularly, PMA differentiated viable from dead cells in cyanobacterial species rather than in other phytoplankton taxa under our experimental conditions. However, our results also showed that it may be necessary to adjust various conditions affecting PMA treatment efficiency to expand its applicability to other phytoplankton. Although all factors contributing to the effects of PMA could not be evaluated, our study showed the applicability of PMA-based molecular approaches, which can be convenient quantitative methods for distinguishing living from dead phytoplankton in freshwater ecosystems. Setting optimal treatment conditions for other phytoplankton species may increase the efficacy of PMA-based molecular approaches.