Abstract

Duckweeds (Lemnaceae species) are extensively used models in ecotoxicology, and chlorophyll fluorescence imaging offers a sensitive and high throughput platform for phytotoxicity assays with these tiny plants. However, the vast number of potentially applicable chlorophyll fluorescence-based test endpoints makes comparison and generalization of results hard among different studies. The present study aimed to jointly measure and compare the sensitivity of various chlorophyll fluorescence parameters in Spirodela polyrhiza (giant duckweed) plants exposed to nickel, chromate (hexavalent chromium) and sodium chloride for 72 h, respectively. The photochemistry of Photosystem II in both dark- and light-adapted states of plants was assessed via in vivo chlorophyll fluorescence imaging method. Our results indicated that the studied parameters responded with very divergent sensitivity, highlighting the importance of parallelly assessing several chlorophyll fluorescence parameters. Generally, the light-adapted parameters were more sensitive than the dark-adapted ones. Thus, the former ones might be the preferred endpoints in phytotoxicity assays. Fv/Fm, i.e., the most extensively reported parameter literature-wise, proved to be the least sensitive endpoint; therefore, future studies might also consider reporting Fv/Fo, as its more responsive analogue. The tested toxicants induced different trends in the basic chlorophyll fluorescence parameters and, at least partly, in relative proportions of different quenching processes, suggesting that a basic distinction of water pollutants with different modes of action might be achievable by this method. We found definite hormetic patterns in responses to several endpoints. Hormesis occurred in the concentration ranges where the applied toxicants resulted in strong growth inhibition in longer-term exposures of the same duckweed clone in previous studies. These findings indicate that changes in the photochemical efficiency of plants do not necessarily go hand in hand with growth responses, and care should be taken when one exclusively interprets chlorophyll fluorescence-based endpoints as general proxies for phytotoxic effects.

Highlights

  • Duckweed species are widely used test objects in ecotoxicology research

  • Chlorophyll fluorescence imaging-based duckweed phenotyping offers a non-destructive, fast, and easy way to assess the potential phytotoxicity of water pollutants, resulting in high throughput systems

  • Our results indicated that different chlorophyll fluorescence induction parameters responded with very different sensitivity to the applied treatments

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Summary

Introduction

Duckweed species (members of the Lemnaceae family) are widely used test objects in ecotoxicology research. Scleid.) in standardized ecotoxicity tests (e.g., OECD Guideline 221, ISO No 20079 and ISO/NP No 20227, respectively). They owe this popularity due to their small size, fast and predominantly vegetative reproduction, simple anatomy and sensitivity to various aquatic toxicants [1,2]. In duckweed-based phytotoxicity assays, toxic effects have been usually quantified via the inhibition of biomass growth in cultures, in terms of frond number, frond area, fresh or dry mass change, respectively [4,5]. The most common ISO [4] and OECD [5] test protocols use sevenday-long exposures and require a control biomass doubling time shorter than 2.5 days in order to ensure adequate sensitivity of tests

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