Because of its reliable results with relatively low cost and labor, this technique has become one of the most widely used methods in ecological studies, including biodiversity assessment ( Valentini et al., 2016 Bylemans et al., 2018), the detection of invasive species ( Borrell et al., 2017 Klymus et al., 2017), and feeding ecology ( Iwanowicz et al., 2016 Yoon et al., 2017 Hawlitschek et al., 2018). Thus, the metabarcoding analysis of ichtyoplankton using the newly designed FishU primers is a promising tool for obtaining useful data to understand the reproduction of fish, such as spawning sites, reproductive periods, population structures, feeding ecology, and diet.ĭNA metabarcoding is a recently established technique that makes it possible to conduct taxonomic identification of entire assemblages in environmental samples via high-throughput sequencing ( Yu et al., 2012). The metabarcoding results by FishU primers successfully presented the diversity of ichthyoplankton directly from the zooplankton net samples in the East/Japan Sea, presenting more accurate and plentiful species numbers than those by MiFish primers. Based on both in silico and in vitro analyses, the newly designed FishU primers outperformed the two previously reported fish-specific universal primer sets (ecoPrimer and MiFish) in taxon coverage, specificity, and accuracy in species identification. First, a piscine-specific universal primer set (FishU) was designed to amplify the region, flanking the highly conserved mitochondrial 12S and 16S ribosomal genes, and it was optimized for the MiSeq platform. Here, a metabarcoding strategy was employed as an alternative to a conventional ichthyoplankton survey, which requires a considerable amount of time, labor, and cost. The spatiotemporal distribution of fish larvae and eggs is fundamental for their reproduction and recruitment in aquatic ecosystems. 5Department of Marine Biology, Pukyong National University, Busan, South Korea.4School of Earth Sciences and Environmental Engineering, Gwangju Institute of Science and Technology, Gwangju, South Korea.3Department of Marine Bioscience, Gangneung-Wonju National University, Gangneung, South Korea.2Aquatic Living Resources Division, FIRA (Korea Fisheries Resources Agency), Busan, South Korea.1Interdisciplinary Program of Biomedical, Mechanical and Electrical Engineering, Pukyong National University, Busan, South Korea.Ah Ran Kim 1, Tae-Ho Yoon 1,2, Chung Il Lee 3, Chang-Keun Kang 4 and Hyun-Woo Kim 1,5*
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