SEAFWA 2017

AUTHORS: Wilawan Thongda, Lauren Davis, Honggang Zhao – School of Fisheries, Aquaculture, and Aquatic Sciences, Auburn University; Chao Li, Marine Science and Engineering College, Qingdao Agricultural University; Benjamin H. Beck, Aquatic Animal Health Research Unit USDA ARS; Eric Peatman, School of Fisheries, Aquaculture, and Aquatic Sciences, Auburn University

ABSTRACT: Black basses (Micropterus spp.) are among the most important sport-fishes in the United States, which has resulted in extensive stockings outside of their native ranges. Interspecific hybridization can lead to the loss of genetic integrity of endemic species. As such, species-diagnostic markers for black basses are important means for monitoring hybridization and maintaining genetic integrity among these species. The aims of this study are to identify species-specific single nucleotide polymorphisms (SNP) markers using genotyping-by-sequencing (GBS) and subsequently, to develop species-diagnostic SNP panels using an Agena MassARRAY system to clearly delineate species and their hybrids among black basses. A total of 190 black basses including Northern Largemouth Bass (M. salmoides), Florida Largemouth Bass (M. floridanus), Redeye Bass (M. coosae), Alabama Bass (M. henshalli), Northern Spotted Bass (M. punctulatus), Shoal Bass (M. cataractae), Smallmouth Bass (M. dolomieu), Guadeloupe Bass (M. treculii), hybrid bass, and intergrade bass were processed for GBS to identify SNP markers. A total of 22665 SNPs were called using a non-reference GBS Universal Network Enabled Analysis Kit (UNEAK) pipeline. Subsequently, we utilized the Agena MassARRAY to validate 29- and 38-plex panels of SNPs selected from the GBS dataset. A total of 494 samples were tested with these two panels, with results demonstrating accurate species identification and revealing varying degrees of hybridization. STRUCTURE and phylogenetic analyses have been employed to capture the population structure and evolutionary relationships among black bass species. This is the first report of SNP markers as a molecular tool to identify the species of black basses.