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'r {] (tv <~lc)~ L;",' u "''''_ TRANSACTIONS OFT H E AM E R)I CAN F ISH E R IE S SO C lET Y ( I Volume 123 July 1994 Number 4 Transactions a/the American Fisheries Society 123:449-459, 1994 @ Copyright by the American Fisheries Society 1994 Nest-Specific DNA Fingerprints of Smallmouth Bass in Lake Opeongo, Ontario MARK L. GROSS AND ANNE R, KApUSCINSKI Department of Fisheries and Wildlife, University of Minnesota St, Paul, Minnesota 55108. USA ANTHONY J. FARAS Institute of Human Genetics, University of Minnesota Minneapolis, Minnesota 55455. USA Abstract,-Methods of DNA fingerprinting were employed to search for nest-specific markers in the population of smallmouth bass Micropterus dolomieu in Lake Opeongo, Ontario, Banding patterns produced from combinations of six restriction endonucleases and seven DNA fingerprint probes were evaluated, Each enzyme and probe combination detected very few polymorphic loci among fry from the same nest. However, comparisons of fingerprints between fry from different nests revealed significant differences in banding patterns, indicating that each nest had a unique fingerprint. Fry from 15 nests in the Jones Bay area of the lake were fingerprinted by using the restriction enzyme Hae III and the probe (GACAk A double-blind test was conducted to deter- mine the ability to assign an unknown fry correctly to its nest of origin and demonstrate that nest- specific fingerprints exist. All fry tested were assigned correctly to their nest of origin by visual comparison of their fingerprints with each of the nest fingerprints, thus demonstrating that nest- specific DNA fingerprints can be generated, Recent advances in molecular genetics have led to the development of a powerful tool, DNA fin- gerprinting, that produces highly variable, genet- ically distinct marks for individuals (e,g" Fields et al. 1989). These individual-specific marks or <<fingerprints" have provided a new source of ge- netic markers that have a variety of applications including studies involving forensics (Bar and Hummel 1991), parentage (Westneat 1990; Rico et al. 1991), linkage analysis (Jeffreys et al. 1986), and genetic diversity (Reeve et al. 1990; Schartl et al. 1991; Turner et al. 1991), In addition to producing individual-specific marks, researchers have begun exploring the use of this technique to generate family and population-specific finger- prints (Gilbert et al. 1990; Wirgin et al. 1991), Family-specific DNA fingerprints would permit monitoring of families in situations where physi- cal tagging or identification is not feasible, a com- mon problem in studies involving fish due to small size of the fish or to logistical problems posed by the behavior of individual fish, Because of their spawning behavior, small- mouth bass Micropterus d%mieu provide an ex- cellent opportunity to search for family (nest)-spe- cific DNA fingerprints in a fish species, Themales build conspicuous nests and defend their off- spring, who initially remain together in a cluster 449 '1",1. , J 1