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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
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Pterois miles Species Pterois volitans10 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
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P. miles P. volitansMixedInvasive Pterois sp.
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P. miles 01
P. miles 06
P. miles 03
P. miles 04
Aquarium 04, 09, 14, 21
Aquarium 27, 28
Aquarium 01, 02, 08, 24, 25
P. miles 02
P. miles 05
P. miles 08
P. miles 07
Aquarium 03, 05, 06, 10,11, 16, 17, 18, 22, 23, 26Long Island 01, 03
Aquarium 19
Aquarium 13
Aquarium 15
Aquarium 20
Aquarium 07
P. volitans 01
P. volitans 02
P. volitans 03
P. antennata 01
P. antennata 02
P. mombasae 01
P. mombasae 02
D. zebra 01
D. zebra 02
P. radiata 01
P. radiata 02
D. brachypterus 01
D. brachypterus 03
D. brachypterus 02
H. hilgendorfii
0.005 substitutions/site
Pterois milesClade
Pterois volitansClade
■ Meristic ID P. miles■ Meristic ID P. volitans■ Meristic ID Unknown
100
100
100
85
87
10077
100
100
10099
94
P. miles 01
P. miles 06
P. miles 03
P. miles 04
Aquarium 04, 09, 14, 21
Aquarium 27, 28
Aquarium 01, 02, 08, 24, 25
P. miles 02
P. miles 05
P. miles 08
P. miles 07
Aquarium 03, 05, 06, 10,11, 16, 17, 18, 22, 23, 26Long Island 01, 03
Aquarium 19
Aquarium 13
Aquarium 15
Aquarium 20
Aquarium 07
P. volitans 01
P. volitans 02
P. volitans 03
P. antennata 01
P. antennata 02
P. mombasae 01
P. mombasae 02
D. zebra 01
D. zebra 02
P. radiata 01
P. radiata 02
D. brachypterus 01
D. brachypterus 03
D. brachypterus 02
H. hilgendorfii
0.005 substitutions/site
Pterois milesClade
Pterois volitansClade
■ Meristic ID P. miles■ Meristic ID P. volitans■ Meristic ID Unknown
100
100
100
85
87
10077
100
100
10099
94
P. miles 01
P. miles 06
P. miles 03
P. miles 04
Aquarium 04, 09, 14, 21
Aquarium 27, 28
Aquarium 01, 02, 08, 24, 25
P. miles 02
P. miles 05
P. miles 08
P. miles 07
P. miles 01
P. miles 06
P. miles 03
P. miles 04
Aquarium 04, 09, 14, 21
Aquarium 27, 28
Aquarium 01, 02, 08, 24, 25
P. miles 02
P. miles 05
P. miles 08
P. miles 07
Aquarium 03, 05, 06, 10,11, 16, 17, 18, 22, 23, 26Long Island 01, 03
Aquarium 03, 05, 06, 10,11, 16, 17, 18, 22, 23, 26Long Island 01, 03
Aquarium 19
Aquarium 13
Aquarium 15
Aquarium 20
Aquarium 07
P. volitans 01
P. volitans 02
P. volitans 03
P. antennata 01
P. antennata 02
P. mombasae 01
P. mombasae 02
D. zebra 01
D. zebra 02
P. radiata 01
P. radiata 02
D. brachypterus 01
D. brachypterus 03
D. brachypterus 02
H. hilgendorfii
0.005 substitutions/site
Pterois milesClade
Pterois volitansClade
■ Meristic ID P. miles■ Meristic ID P. volitans■ Meristic ID Unknown
100
100
100
85
87
10077
100
100
10099
94
Current Data
Disagreement between mitochondrial DNA and meristic identities
14 nuclear DNA loci lack species specific variation for P. miles and P. volitans
Act-2 AldB Cam-3
CK CKA7 ChymB-6
ITS LDH LDHA6
Mlc-3 Ops-1 RP1
RP2 SSU
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Objective I
Investigate hybridization
using Randomly Amplified
Polymorphic DNAs (RAPDs) to
develop a nuclear DNA marker
to distinguish between Pterois
miles and Pterois volitans
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Methods
Randomly Amplified Polymorphic DNAs (RAPDs)
Primer sitesCopied DNA fragments
Template DNA
1. Polymerase Chain Reaction5’ 3’
1 2 3 4
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A ? B
2. Gel Electrophoresis
longest fragments
shortest fragments
Size marke
r
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Objective II
Examine the genetic diversity
of North Carolina lionfish to
determine their source
population
?
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Methods
P. miles 01
P. miles 06
P. miles 03
P. miles 04
Aquarium 04, 09, 14, 21
Aquarium 27, 28
Aquarium 01, 02, 08, 24, 25
P. miles 02
P. miles 05
P. miles 08
P. miles 07
Aquarium 03, 05, 06, 10,11, 16, 17, 18, 22, 23, 26Long Island 01, 03
Aquarium 19
Aquarium 13
Aquarium 15
Aquarium 20
Aquarium 07
P. volitans 01
P. volitans 02
P. volitans 03
P. antennata 01
P. antennata 02
P. mombasae 01
P. mombasae 02
D. zebra 01
D. zebra 02
P. radiata 01
P. radiata 02
D. brachypterus 01
D. brachypterus 03
D. brachypterus 02
H. hilgendorfii
0.005 substitutions/site
Pterois milesClade
Pterois volitansClade
■ Meristic ID P. miles■ Meristic ID P. volitans■ Meristic ID Unknown
100
100
100
85
87
10077
100
100
10099
94
P. miles 01
P. miles 06
P. miles 03
P. miles 04
Aquarium 04, 09, 14, 21
Aquarium 27, 28
Aquarium 01, 02, 08, 24, 25
P. miles 02
P. miles 05
P. miles 08
P. miles 07
Aquarium 03, 05, 06, 10,11, 16, 17, 18, 22, 23, 26Long Island 01, 03
Aquarium 19
Aquarium 13
Aquarium 15
Aquarium 20
Aquarium 07
P. volitans 01
P. volitans 02
P. volitans 03
P. antennata 01
P. antennata 02
P. mombasae 01
P. mombasae 02
D. zebra 01
D. zebra 02
P. radiata 01
P. radiata 02
D. brachypterus 01
D. brachypterus 03
D. brachypterus 02
H. hilgendorfii
0.005 substitutions/site
Pterois milesClade
Pterois volitansClade
■ Meristic ID P. miles■ Meristic ID P. volitans■ Meristic ID Unknown
100
100
100
85
87
10077
100
100
10099
94
P. miles 01
P. miles 06
P. miles 03
P. miles 04
Aquarium 04, 09, 14, 21
Aquarium 27, 28
Aquarium 01, 02, 08, 24, 25
P. miles 02
P. miles 05
P. miles 08
P. miles 07
P. miles 01
P. miles 06
P. miles 03
P. miles 04
Aquarium 04, 09, 14, 21
Aquarium 27, 28
Aquarium 01, 02, 08, 24, 25
P. miles 02
P. miles 05
P. miles 08
P. miles 07
Aquarium 03, 05, 06, 10,11, 16, 17, 18, 22, 23, 26Long Island 01, 03
Aquarium 03, 05, 06, 10,11, 16, 17, 18, 22, 23, 26Long Island 01, 03
Aquarium 19
Aquarium 13
Aquarium 15
Aquarium 20
Aquarium 07
P. volitans 01
P. volitans 02
P. volitans 03
P. antennata 01
P. antennata 02
P. mombasae 01
P. mombasae 02
D. zebra 01
D. zebra 02
P. radiata 01
P. radiata 02
D. brachypterus 01
D. brachypterus 03
D. brachypterus 02
H. hilgendorfii
0.005 substitutions/site
Pterois milesClade
Pterois volitansClade
■ Meristic ID P. miles■ Meristic ID P. volitans■ Meristic ID Unknown
100
100
100
85
87
10077
100
100
10099
94
ABI 3100 Genetic Analyzer
NC lionfish
Phylogenetic tree or Cluster
analysis
?
PCR & Sequence Reaction
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Questions?
Leo (NC lionfish) at CMS
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Mer = mMito = vNuc = v
Mer = vMito =
mNuc = m
Mer = mMito = vNuc = m
Mer = vMito =
mNuc = v
Nuclear and mitochondrial
DNA disagreement
-Hybridization-Incomplete
lineage sorting
Meristic and DNA disagreement
Meristic characters not fixed for P. miles
and P. volitans
Mer = meristic countsMito = mitochondrial DNANuc = nuclear DNA
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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.