Effectiveness of Lugol’s iodine solution for long-term preservation of zooplankton samples for molecular analysis

Abstract

Molecular analysis is a common tool for marine ecological and biological research. For genomic analysis of zooplankton, ethanol preservation and freezing are often used to preserve samples until analysis. However, these methods have disadvantages, such as the loss of morphological information. Recently, 10% Lugol’s iodine solution (10% Lugol) has been shown to be an effective preservative of plankton samples for molecular analysis, even after 18 months. However, that study only reported a PCR-based molecular study using copepods. We tested the preservation effectiveness of 10% Lugol on various zooplankton over 33 months by comparing them to samples preserved by freezing or in 5% formalin seawater. The results revealed that the total amounts of DNA extracted from crustacean and gelatinous zooplankton preserved in 10% Lugol were the same or higher than those preserved in formalin or by freezing. Gel electrophoresis of the extracted DNA indicated that the DNA of the samples preserved in 10% Lugol was not fragmented during the preservation period. PCR amplification of a partial 18S rRNA gene using DNA extracted from various zooplankton taxa (siphonophores, copepods, ostracods, doliolids, polychaetes, pteropods, euphausiids, and chaetognaths) was successful. Sequences of morphologically identified species preserved in 10% Lugol had BLAST hits to sequences of these species deposited in Genbank, with a similarity of 100%, which indicated there was no sequence alteration during the preservation period. Thus, we conclude that 10% Lugol is a suitable preservative for molecular analysis of various zooplankton taxonomic groups.