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), Chlorophyta, 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.
Highlights
Discrimination between viable and dead phytoplankton cells can provide novel perspectives in ecological research [1,2,3]
Propidium monoazide (PMA) treatment has been used in microbiological studies, especially for detecting viable bacteria, but a few studies have applied PMA dye for phytoplankton in freshwater ecosystems
Our results showed that application of PMA for phytoplankton in freshwater can potentially distinguish viable from dead cells as in microbial studies
Summary
Discrimination between viable and dead phytoplankton cells can provide novel perspectives in ecological research [1,2,3]. In terms of energy flows, viability of phytoplankton could serve as one of the main factors determining seston food quality for zooplankton [4,5,6]. Some calanoids, such as Eucalanus pileatus, can discriminate between the food quality of different particles and prefer living rather than dead phytoplankton cells [7, 8]. Detection of viable phytoplankton species is essential to prevent the spread of non-indigenous. Applicability of PMA for distinguishing living phytoplankton cells species transported in the ballast water of ships [9, 10]. Various methods have been tested to reduce the number of viable organisms in ballast water, and the potential survival and regrowth of harmful organisms should be measured after treatment in accordance with the guidelines of the Ballast Water Convention adopted by the International Maritime Organization (IMO) [11,12,13]
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