• ALGAE
• /
• 제34권1호
• /
• pp.57-70
• /
• 2019

DNA metabarcoding is currently used for large-scale taxonomic identification to understand the community composition in various marine ecosystems. However, before being widely used in this emerging field, this experimental and analytic approach still has several technical challenges to overcome, such as polymerase chain reaction (PCR) bias, and lack of well-established metabarcoding markers, a task which is difficult but not impossible to achieve. In this study, we present an adapted PCR-free small-organelles enriched metagenomics (SoEM) method for marine biodiversity assessment. To avoid PCR bias and random artefacts, we extracted target DNA sequences without PCR amplification from marine environmental samples enriched with small organelles including mitochondria and plastids because their genome sequences provide a valuable source of molecular markers for phylogenetic analysis. To experimentally enrich small organelles, we performed subcellular fractionation using modified differential centrifugation for marine environmental DNA samples. To validate our SoEM method, two marine environmental samples from the coastal waters were tested the taxonomic capturing capacity against that of traditional DNA metabarcoding method. Results showed that, regardless of taxonomic levels, at least 3-fold greater numbers of taxa were identified in our SoEM method, compared to those identified by the conventional multi-locus DNA metabarcoding method. The SoEM method is thus effective and accurate for identifying taxonomic diversity and presents a useful alternative approach for evaluating biodiversity in the marine environment.

• ALGAE
• /
• 제34권4호
• /
• pp.303-313
• /
• 2019

Floating seaweed rafts are a surface-pelagic habitat that serve as substrates for benthic flora and fauna. Since 2008, Sargassum horneri clumps have periodically invaded the Korea Strait. In this study, the polymerase chain reaction-free small-organelles enriched metagenomics method was adopted to identify the species of epibiotic eukaryotes present in floating S. horneri fronds. A total of 185 species were identified, of which about 63% were previously undetected or unreported in Korean waters. The rafts harbored a diverse assemblage of eukaryotic species, including 39 Alveolata, 4 Archaeplastida, 95 Opisthokonts, 4 Rhizaria, and 43 Stramenopiles. Of these 185 taxa, 48 species were found at both Sargassum rafts collection stations and included 24 Stramenopiles, 17 Alveolata, and 7 Opisthokonts. Among these, the highest proportion (50%) of species was photo-autotrophic in basic trophic modes, while the proportion of phagotrophic, osmo- or saprotrophic, and parasitic modes were 43.8%, 4.2%, and 2.1%, respectively. This study demonstrates the contribution of floating Sargassum rafts as dispersal vectors that facilitate the spread of alien species.