Abstract
Phytoplankton are primary producers and can be important indicators of environmental change. To monitor the plankton species
composition of environmental seawater samples, we developed a molecular method composed of colony polymerase chain reaction
(PCR), polymerase chain reaction–restriction fragment length polymorphism (PCR-RFLP), and sequencing. A clone library of the
ribosomal small subunit RNA gene (18S rDNA) in the nuclear genome was constructed by environmental PCR using a newly designed
primer set and clones were directly amplified by colony PCR. To select unique putative clones, we choose a PCR-RFLP method
that employed two restriction enzymes (MseI and Tsp509I). After the PCR-RFLP pattern was evaluated, selected clones were sequenced and analyzed. In this study, we revealed the
hidden biodiversity in environmental seawater containing a wide range of taxonomic groups in the Alveolata (Ciliphora and
Dinophyceae), Euglenozoa, Stramenopiles (Bacillariophyta), and Viridiplantae (Chlorophyta) without the need to conduct extensive
colony isolation techniques. Moreover, we found species of fungi and Metazoa (Arthropoda, Annelida, and Mollusca). Therefore,
this improved molecular method can be used to generate a robust database describing the species diversity of environmental
samples and provide useful information regarding the dynamics of the eukaryotic plankton community structure.
composition of environmental seawater samples, we developed a molecular method composed of colony polymerase chain reaction
(PCR), polymerase chain reaction–restriction fragment length polymorphism (PCR-RFLP), and sequencing. A clone library of the
ribosomal small subunit RNA gene (18S rDNA) in the nuclear genome was constructed by environmental PCR using a newly designed
primer set and clones were directly amplified by colony PCR. To select unique putative clones, we choose a PCR-RFLP method
that employed two restriction enzymes (MseI and Tsp509I). After the PCR-RFLP pattern was evaluated, selected clones were sequenced and analyzed. In this study, we revealed the
hidden biodiversity in environmental seawater containing a wide range of taxonomic groups in the Alveolata (Ciliphora and
Dinophyceae), Euglenozoa, Stramenopiles (Bacillariophyta), and Viridiplantae (Chlorophyta) without the need to conduct extensive
colony isolation techniques. Moreover, we found species of fungi and Metazoa (Arthropoda, Annelida, and Mollusca). Therefore,
this improved molecular method can be used to generate a robust database describing the species diversity of environmental
samples and provide useful information regarding the dynamics of the eukaryotic plankton community structure.
- Content Type Journal Article
- DOI 10.1007/s10811-010-9509-7
- Authors
- Sang-Rae Lee, Pusan National University Marine Research Institute Busan 609-735 South Korea
- Jung Hyun Oak, Pusan National University Marine Research Institute Busan 609-735 South Korea
- Ik Kyo Chung, Pusan National University Division of Earth Environmental System Busan 609-735 South Korea
- Jin Ae Lee, Inje University School of Environmental Science and Engineering Gimhae 621-749 South Korea
- Journal Journal of Applied Phycology
- Online ISSN 1573-5176
- Print ISSN 0921-8971
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