1. 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

2. 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

3. P. miles
P. volitans
Mixed
Invasive Pterois sp.

4. 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

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

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

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

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

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

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

11. 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

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