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Evaluation of the 18S rRNA clone library approach to study the diversity of the macroeukaryotic leaf-epiphytic community of the seagrass Posidonia oceanica (L.) Delile

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Abstract

The sequence comparisons among genes codifying for the RNA component of the small ribosomal subunit (16S rRNA or 18S rRNA) in cellular organisms have been largely used to reconstruct their phylogenies, and hence the identification of taxa by means of a molecular approach. Furthermore, the direct DNA isolation from environmental samples and the PCR amplification of the pool of rRNA genes with the subsequent cloning and sequencing have opened the door to the description of naturally occurring microbial communities independently from any culturing technique or morphological identification. These studies have unveiled an enormous hidden diversity in a wide variety of microbial communities. Our main objective was to evaluate the usefulness of the 18S rRNA gene clone libraries to describe the structure of the macroeukaryotic leaf-epiphytic assemblage of the seagrass Posidonia oceanica, and monitor the changes occurring in different stages of its seasonal succession (winter, spring and summer). To that end, we compared the results of these libraries with those provided by classical microscopy techniques. Among both approaches, the screening of clone libraries rendered the highest number of distinct units named operational phylogenetic units. However, diversity estimates provided by both methods were comparable and rendered the highest Shannon Diversity Index (H′) at the end of the succession. The major discrepancies were on the different occurrence of some groups. For example, macroalgae were the most frequent epiphytes counted by microscopy, whereas metazoa (specially, bryozoa) dominated the clone libraries. Altogether the results indicate that clone libraries constitute an excellent complementary approach to classical microscopy methods. To the best of our knowledge, this is the first attempt to describe a marine macroeukaryotic community using a molecular approach such as the analysis of 18S rRNA gene clone libraries.

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Acknowledgments

We thank the Marine Microbiology Group and the Marine Macrophyte Ecology Group from IMEDEA for facilities and help with the field and lab work. This study was funded by the Government of the Balearic Islands (Acció Especial de recerca, desenvolupament tecnològic i innovació (2006) and the UGIZC research contract), the Spanish Ministry of Education and Science (Acción Complementaria CTM2005-23775-E) and the Spanish Ministry of Science and Innovation (research projects CLG2006-12714-C02-02 and Consolider-Ingenio 2010 CE-CSD2007-0005). F.J.M.P thanks ‘Conselleria de Economia, Hisenda i Innovació’ of Balearic Government for a PhD grant (FPI05) that supported this work. All experiments done in this study comply with the current laws of Spain.

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Authors and Affiliations

  1. Marine Macrophyte Ecology Group, Ecology and Marine Resources Department, IMEDEA (CSIC-UIB), Miquel Marqués 21, 07190, Esporles, Majorca, Spain

    F. J. Medina-Pons & J. Terrados

  2. Marine Microbiology Group, Ecology and Marine Resources Department, IMEDEA (CSIC-UIB), Miquel Marqués 21, 07190, Esporles, Majorca, Spain

    A. López-López, P. Yarza & R. Rosselló-Móra

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  1. F. J. Medina-Pons
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  2. J. Terrados
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Correspondence to F. J. Medina-Pons.

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Communicated by T. Reusch.

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Medina-Pons, F.J., Terrados, J., López-López, A. et al. Evaluation of the 18S rRNA clone library approach to study the diversity of the macroeukaryotic leaf-epiphytic community of the seagrass Posidonia oceanica (L.) Delile. Mar Biol 156, 1963–1976 (2009). https://doi.org/10.1007/s00227-009-1221-2

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