A DNA mini-barcoding system for endangered unionid mussels in the Lake Biwa system in Japan
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TECHNICAL NOTE
A DNA mini‑barcoding system for endangered unionid mussels in the Lake Biwa system in Japan Kohji Mabuchi1 · Kazuya Nishida1 Received: 25 January 2019 / Accepted: 16 May 2020 © Springer Nature B.V. 2020
Abstract A mini-barcoding system was developed to help identify endangered unionid mussels in the Lake Biwa system in Japan. A single primer set was designed to amplify a ca. 140-bp barcode fragment based on the published mitochondrial 16S rRNA gene sequences (ca. 350 bp) of Japanese unionids. The new primers successfully amplified genomic DNA from the foot tissues of 81 specimens of 12 target taxa from the Lake Biwa region. Sequencing, BLAST-searches and phylogenetic analyses of the amplified fragments confirmed that the mini-barcode had the same identification ability as the original sequences for Lake Biwa taxa. The mini-barcoding system also obtained the short barcodes from excreted materials, indicating its usefulness in nondestructive identification of these mussels. Keywords DNA barcode · Unionidae · 16S rRNA · Lake biwa Unionid freshwater mussels are burrowing, filter-feeding bivalves that occur worldwide. They reside anywhere from small ditches to lakes, influencing macroinvertebrate assemblage structure in streams (Vaughn and Spooner 2006). Before the alteration and degradation of freshwater habitats, they were one of the most abundant bivalves in freshwater, but many of them are now declining in many countries (Carella et al. 2016). Unionid populations in Japan have also declined recently, and some have gone extinct widely there as irrigation systems have been rebuilt (Kondo 2008). Lake Biwa, located in central Japan, is one of the world’s oldest lakes, dating to at least 4 million years ago (Kawabe 1989). The lake and its affluent and effluent rivers harbor 66 endemic taxa, including 7 unionid bivalves (Nishino 2017). Although their specific or subspecific status and scientific names are debated (e.g. Sano et al. 2017), Kihira et al. (2003) recognized 13 unionid taxa in the lake and its catchment, and the Prefectural Red Data Book (Scientific Electronic supplementary material The online version of this article (https://doi.org/10.1007/s12686-020-01160-w) contains supplementary material, which is available to authorized users. * Kohji Mabuchi [email protected] 1
Lake Biwa Branch Office, National Institute for Environmental Studies, 5‑34 Yanagasaki, Otsu, Shiga, Japan
Committee for Research into the Wildlife in Shiga Prefecture, 2016) regarded 11 of the 13 unionids as endangered taxa (Table 1). The unionid mussels in the Lake Biwa system are used as spawning sites by ten native freshwater fish species with some host-parasite specificity: seven Acheilognathinae and three Sarcocheilichthys (Kihira et al. 2003). Consequently, together with their function as ecosystem engineers, the presence and diversity of unionid mussels are important for the conservation of biodiversity in the system directly and indirectly. Despite their conservation importance, their morphological ident
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