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A protocol for conducting 7-day daily renewal tests with Lemna gibba

Nature Protocols volume 2, pages 979–987 (2007)Cite this article

Abstract

Lemna gibba (a duckweed) is a freshwater macrophyte commonly used in toxicity testing, and Lemna spp are currently the only aquatic higher plants required for evaluation of pesticides under the pesticide registration guidelines of the EPA. The methods currently available for toxicity testing by various organizations and agencies, including ASTM, OECD, EPA and Environment Canada, are largely static or semistatic tests with unspecified renewal intervals (OECD) and may not provide a consistent means of exposure owing to short toxicant half-life in aquatic media, uptake of chemical by plants and evaporation of nutrient media. The procedure outlined here details a simple and efficient 7-day daily static renewal procedure for conducting toxicity tests with L. gibba, the appropriate end points to assess, the statistical criteria necessary for analyzing the toxicity data, as well as the steps required to culture and maintain L. gibba. This protocol is based on a modified version of a widely accepted static method.

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Figure 1: Visual reference for L. gibba test cultures.
Figure 2: Visual reference for the experimental setup and renewal procedure for a 7-day daily static renewal test with L. gibba using a randomized complete block design (seven treatments, including solvent controls, nonsolvent controls and five treatment levels, and three replicates) on a tray in a laminar flow hood.
Figure 3: Visual reference for conducting multiple 7-day daily static renewal tests with L. gibba, examples of potential phytotoxic symptoms for different stressor types, and an example of a concentration–response effects sequence.

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Acknowledgements

We acknowledge Dr Bruce Greenberg and Dr Mark Hanson.

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Authors and Affiliations

  1. Department of Environmental Studies, Center for Reservoir and Aquatic Systems Research, Baylor University, One Bear Place #97388, Waco, 76798-7388, Texas, USA

    Richard A Brain

  2. Department of Environmental Biology, Centre for Toxicology, University of Guelph, Guelph, N1G 2W1, Ontario, Canada

    Keith R Solomon

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Correspondence to Richard A Brain.

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Brain, R., Solomon, K. A protocol for conducting 7-day daily renewal tests with Lemna gibba. Nat Protoc 2, 979–987 (2007). https://doi.org/10.1038/nprot.2007.146

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