Morphological and genetic identification of formalin-fixed gobioid larvae and description of postflexion larvae of Parag
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Morphological and genetic identification of formalin‑fixed gobioid larvae and description of postflexion larvae of Paragunnellichthys sp. and Ctenogobiops feroculus Nozomi Hanahara1 · Kei Miyamoto1 · Shin‑ichiro Oka1 Received: 30 March 2020 / Revised: 13 July 2020 / Accepted: 15 July 2020 © The Ichthyological Society of Japan 2020
Abstract Gobioid larvae collected from the coast of Okinawa Island, southern Japan, were identified on the basis of a combination of morphological characters and a sequence of a hypervariable region of mitochondrial 12S ribosomal RNA (12S rRNA) gene (138–145 base pairs). A short-term formalin fixation technique enabled identification using both morphological and genetic methods. Thirteen of the 21 types of gobioid larvae assessed were identified to the species level. Additionally, we described the morphology of the postflexion larvae of wormfish Paragunnellichthys sp. and shrimp-associated goby Ctenogobiops feroculus, identified for the first time in their respective genera. Keywords 12S rRNA · DNA barcoding · Molecular identification · Fish larvae · Goby
Introduction Gobioid fish, suborder Gobioidei (Teleostei), is an important constituent of larval fish assemblages in tropical and subtropical coastal waters (e.g. Clarke 1991; Maeda and Tachihara 2014; Isari et al. 2017). Nevertheless, the understanding of early life history of gobies is poorly known (Borges et al. 2011). This lack of current knowledge can largely be attributed to the fact that gobioid larvae are difficult to identify. Identification of fish larvae has been traditionally based on morphological characters, which are often not sufficient to identify larval fish to the species level (Ko et al. 2013). This article was registered in the Official Register of Zoological Nomenclature (ZooBank) as 3CCEE403-456B-473B-A4A8061E086CB780. This article was published as an Online First article on the online publication date shown on this page. The article should be cited by using the doi number. Electronic supplementary material The online version of this article (https://doi.org/10.1007/s10228-020-00769-z) contains supplementary material, which is available to authorized users. * Nozomi Hanahara [email protected] 1
Okinawa Churashima Foundation Research Center, 888 Ishikawa, Motobu‑cho, Okinawa 905‑0206, Japan
Recently, genetic methods called DNA barcoding have been used to identify fish eggs and larvae (e.g. Victor et al. 2009; Kawakami et al. 2010; Marancik et al. 2010; Kimmerling et al. 2018). This technique can distinguish even closely related species (e.g. Baldwin et al. 2009; Tawa et al. 2014). These genetic methods enable any researcher with expertise in molecular biology to achieve results of similar precision. However, there are certain limitations in identifying fish eggs and larvae by genetic methods such as mitochondrial introgression, the lack of reference sequences in the database, and misidentification of reference data (Kawakami et al. 2010; Leis 2015). Therefore, it is preferable to i
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