Blenniiformes diversity from an environmental DNA (eDNA) pilot project in the Banggai MPA, Central Sulawesi, Indonesia

Abstract

Sulawesi and its satellite archipelagos lie within the Wallacea region and Coral Triangle biodiversity “hotspots”. Environmental DNA (eDNA) metabarcoding can help detect easily missed or challenging to identify groups such as blennies (Blenniiformes). A pilot eDNA study within the recently established Banggai MPA aimed to provide fine-scale data. Seawater samples were collected at four sites (3 replicates/site) around Banggai Island. DNA extraction (Bionesia, Bali), metabarcoding (MiFish 12S rRNA primers) and sequence library preparation (Barber Lab, University of California Los Angeles), Nextseq High-throughput sequencing, and Anacapa Toolkit analysis produced 254,847 teleost reads aggregated into amplicon sequence variants (ASVs). At 60%, 90% and 100% Bayesian confidence levels, respectively, 1612, 1610 and 1573 reads were assigned to Blenniiformes, while 15 of 20, 12 of 20 and 11 of 18 ASVs were assigned to species level. Blenniiformes ASVs comprised two families: Blenniidae (genera Atrosalarias, Cirripectes, Ecsenius, Exallias, Meiacanthus, Omobranchus, Salarias) and Tripterygiidae (genus Enneapterygius). Assigned genera overlapped with 1998 visual survey data. Phylogenetic analysis (MEGA 11) incorporated NCBI GenBank accessions (BLASTn tool). Blenniiformes community composition differed between sites. This study contributes baseline biodiversity data for Blenniiformes in the Banggai MPA; however, unassigned ASVs and phylogenetic analyses highlight the gaps in 12S rRNA reference databases.