Advisor: Wilson Freshwater The use of DNA markers to identify and explore the genetic diversity of lionfishes Pterois miles and Pterois volitans Rebecca M. Hamner Advisor: Wilson Freshwater

Pterois miles Pterois volitans Species 10 Dorsal Fin Rays 11 6 Anal Fin Rays 7 Shorter Pectoral Fin Size Longer Smaller Spot size Larger Indian Ocean, S. Africa to Red Sea, East to Sumatra Distribution West Pacific Ocean from Japan to Australia, off western Australia, South Pacific to Pitcairn Group

P. miles P. volitans Mixed Invasive Pterois sp.

Current Data Disagreement between mitochondrial DNA and meristic identities 14 nuclear DNA loci lack species specific variation for P. miles and P. volitans As you can see by the color discrepancy in this phylogenetic tree, I previously found disagreement between the meristic and mitochondrial DNA IDs Which led us to investigate hybridization as a possible cause of this conflict Last year I received an Undergrad Research Fellowship to search for a sequence-based nuclear DNA marker to test for hybridization by comparing it to the mito DNA and meristic IDs seq-based DNA markers used to distinguish spp. by differences in actual DNA seq The 14 nuclear DNA loci that I sequenced had no species specific variations that could be used to ID the fishes Act-2 AldB Cam-3 CK CKA7 ChymB-6 ITS LDH LDHA6 Mlc-3 Ops-1 RP1 RP2 SSU

Objective I Investigate hybridization using Randomly Amplified Polymorphic DNAs (RAPDs) to develop a nuclear DNA marker to distinguish between Pterois miles and Pterois volitans

Methods Randomly Amplified Polymorphic DNAs (RAPDs) 1. Polymerase Chain Reaction 5’ 3’ 1 2 3 4 Template DNA Primer sites Copied DNA fragments

2. Gel Electrophoresis A ? B longest fragments shortest fragments Size marker A ? B longest fragments shortest fragments

Objective II Examine the genetic diversity of North Carolina lionfish to determine their source population ?

PCR & Sequence Reaction Phylogenetic tree or Cluster analysis Methods ? NC lionfish PCR & Sequence Reaction ABI 3100 Genetic Analyzer Phylogenetic tree or Cluster analysis

Leo (NC lionfish) at CMS Questions? Leo (NC lionfish) at CMS

Meristic and DNA disagreement Mer = m Mito = v Nuc = v Meristic and DNA disagreement Meristic characters not fixed for P. miles and P. volitans Mer = m Mito = v Nuc = m Mer = v Mito = m Nuc = m Nuclear and mitochondrial DNA disagreement -Hybridization -Incomplete lineage sorting Mer = v Mito = m Nuc = v Mer = meristic counts Mito = mitochondrial DNA Nuc = nuclear DNA

2. Gel Electrophoresis Partis, L. and R. J. Wells. 1996. Identification of fish species using random amplified polymorphic DNA (RAPD). Molecular and Cellular Probes, 10: 435-441.