A Multigene Phylogeny of
Syngnathid Fishes
Healy HamiltonCalifornia Academy
of Sciences
Norah SaarmanUniversity of California,
Santa Cruz
Beth Moore, Graham Short, & W. Brian SimisonCalifornia Academy of Sciences
Martin GomonMuseum Victoria
David Harasti
Acknowledgments
Dr. Jeff Leis and Mark McGroutherAustralian Museum
Dianne Bray, Museum Victoria
Ralph Foster, South Australian Museum
David Harasti, Dept. of Primary Industries, NSW
Clive Roberts, Carl StruthersMuseum of New Zealand, Te Papa, Tongarewa
Dr. David Neely, Dave Catania, Jens Vindum, Jim EvansCalifornia Academy of Sciences
Dr. E. O. Wiley and Andy BentleyNational History Museum, The University of Kansas
Dr. Mark Erdmann, Conservation International
Dr. Gerry Allen, Western Australian Museum
Institute for Research & Development, New CaledoniaPierre LaBoute, Noumea, New Caledonia
Dr. Robert Browne, Dr. David Muirhead, Kevin SmithInshore Fish Group
Dr. Theodore W. Pietsch and Katherine PearsonUniversity of Washington
SEAHORSE
SLEUTH
Outline: A Multigene Phylogeny of Syngnathid Fishes
1. Introduction: why syngnathids?
2. Issues in current syngnathid taxonomy
3. Molecular approach to address syngnathid systematics
4. Results: molecular phylogenetic hypotheses, mapping of non-molecular characters
5. Conclusions and next steps
Photos:
David
Harasti
Introduction: Why we study syngnathid fishes
• Syngnathid fishes: a diverse, unique and charismatic group
• over 50 genera and at least 278 valid species of pipefishes, seahorses, sea dragons, and pipehorses
DD VU EN CR
Seahorses 23 9 1
Pipefishes 5 1
Seadragons 2
Pipehorses 5
IUCN Syngnathid listings
DD = Data Deficient VU = Vulnerable EN = EndangeredCR = Critically Endangered
Syngnathidae are characterized by:
unique gills
absence of pelvic fins
dermal plates in rings
unusual reproductive strategies, including sex role reversal and male brooding, with complex and varied brood-pouch morphology
Hippocampus whitei
Graham Short
Hippocampus abdominalis
Graham Short
Hippocampus
bargibanti
Graham Short
Hippocampus pontohi
David Harasti
Weedy Sea Dragon
Phyllopteryx taeniolatus
Graham Short
Leafy Sea Dragon
Phycodurus eques
Graham Short
Pygmy pipehorse
Idiotropiscis lumnitzeri
Graham Short
Spotted pipefish
Stigmatopora argus
Graham Short
Briggs Crested pipefish
Histiogamphelus briggsi
Graham Short
Bent stick pipefish
Trachyrhampus
bioarctatus
David Harasti
The Problem: Systematic relationships unresolved
• Unresolved systematic relationships at multiple levels
• Generic and specific boundaries are often uncertain
• evolutionary relationships at the species, genus and subfamily level are largely unresolved
• order Gasterosteiformes / Syngnathiformesdebated
• many new species remain to be described
• Many genera are monotypic, yet no study has measured their degree of evolutionary uniqueness
Conservation and management of syngnathids is undermined by their challenging alpha
taxonomy and poorly resolved systematics
The problem
Only two recognized sub-families, the Hippocampinae and the Syngnathinae
B. Syngnathinae: all others; is
paraphyletic and includes 54
genera and 233 valid species
A. Hippocampinae: seahorses
includes only one genus with
38 - 50 species (debated)
(Paxton 1975)(Whitley, 1952)
• Seadragons (Solegnathinae• Tail-brooding Pipefishes(original Syngnathinae)• Abdominal-brooding Pipefishes(Doryrhamphinae and Nerophinae)• Pipehorses (originally included in Hippocampinae)
Our Approach: Methods
Morphological characters have proven problematic
Molecular studies to date (Wilson et al., 2001 & 2003) suffer from limited taxon and molecular character sampling. WE HAVE ADDED:
20 new genera
• Total of 59 taxa (37 genera)
4 new genetic markers, including 2 nuclear fragments
• 6 markers total, ~4500 aligned bps
• Mitochondrial; CO1, ND1, 12s, 16s
• Nuclear; RP1 intron of S7, Myh 6
Our Approach: Increased Taxon Sampling
Previous Generic Assignment Species Wilson (2003) This Study Previous Generic Assignment Species Wilson (2003) This Study
Acentronura 5 1 Kimblaeus 1
Idiotropiscis 1 1 Leptoichthys 1 1
Anarchopterus2
Leptonotus3
Apterygocampus1
Lissocampus5
Bhanotia3
Maroubra2 1
Bryx5
Micrognathus7 1
Bulbonaricus3
Microphis 21 2
Campichthys4
Minyichthys4
Choeroichthys 6 2 Mitotichthys 4 1
Corythoichthys 10 3 2 Nannocampus 4
Cosmocampus 14 1 Nerophis 3 1 1
Doryichthys 5 1 Notiocampus 1
Doryrhamphus 14 1 3 Penetopteryx 2
Dunkerocampus 1 1 1 Phoxocampus 3
Enneacampus2
Phycodurus1 1
Entelurus 1 1 1 Phyllopteryx 1 1 1
Festucalex 6 1 Pseudophallus 3 1
Filicampus 1 1 Pugnaso 1 1 1
Halicampus 12 1 Siokunichthys 5 1
Haliichthys 1 1 Solegnathus 5 1 2
Heraldia1
Stigmatopora3 4 3
Hippichthys 5 1 Stipecampus 1 1
Hippocampus 45 9 9 Syngnathoides 1 1
Histiogamphelus 2 2 Syngnathus 32 14 3
Hypselognathus 2 1 1 Trachyrhamphus 3 1
Ichthyocampus2
Urocampus2 1 1
Kaupus 1 1 1 Vanacampus 4 2 4
Total 54 genera (16 monotypic) 278 species 16 genera 37 genera
Tailbrooding pipefish
0.2
Corythoichthys amplexus
Maroubra perserrata
Hippocampus fuscus
Stipecampus cristatus
Hippocampus denise
Cosmocampus elucens
Doryichthys boaja
Syngnathus fuscus
Histiogamphelus cristatus
Hippocampus pontohi
Vanacampus poecilolaemus
Fistularia petimba
Hippocampus ingens
Siokunichthys nigrolineatus
Filicampus tigiris
Halicampus sp
Microphis ocellatus
Phycodurus eques
Choeroichthys sculptus
Entelerus aequoreus
Mitotichthys semistriatus
Stigmatopora nigra
Acentronura tentaculata
Doryrhamphus jansii
Nerophis spp
Hypselognathus rostratus
Hippicthys penicillus
Syngnathus louisianae
Stigmatopora argus
Dunckerocampus chapmani
Festucalex cinctus
Aulostomus chinensis
Hippocampus trimaculatus
Hippocampus zosterae
Vanacampus phillipi
Doryrhamphus excises
Corythoichthys intestinalis
Micrognathus andersonii
Solegnathus spinosissimus
Trachyrhamphus bioarctatus
Solegnathus hardwicki
Hippocampus abdominalis
Histiogamphelus briggsi
Pugnaso curtirostris
Solenostomus cyanopterus
Stigmatopora narinosa
Haliichthys taeniophorus
Hippocampus whitei
Kaupus costatus
Vanacampus margaritifer
Syngnathoides biaculeatus
Phyllopteryx taeniolatus
Hippocampus bargibanti
Urocampus carinirostris
Idiotropiscis lumnitzeri
Microphis doecata
Syngnathus exilisPseudophallus brasiliensis
Aulostomus maculatus
Vanacampus vercoi
Choeroichthys brachysoma
Leptoichthys fistularis
Doryrhamphus dactyliophorus
0.66
1
1
1
1
1
0.85
1
0.65
1
0.95
1
1
1
1
0.72
1
1
1
1
1
0.79
0.71
1
1
1
0.52
1
1
1
1
1
1
1
1
0.65
1
1
0.51
1
1
1
1
1
1
1
1
0.99
1
1
1
1
1
1
1
10.91
0.99
1
1
Abdominal-brooding pipefish
Results: Bayesian consensus tree
Seadragons
Pipehorses
Seahorses
Outgroups
0.2
Corythoichthys amplexus
Maroubra perserrata
Hippocampus fuscus
Stipecampus cristatus
Hippocampus denise
Cosmocampus elucens
Doryichthys boaja
Syngnathus fuscus
Histiogamphelus cristatus
Hippocampus pontohi
Vanacampus poecilolaemus
Fistularia petimba
Hippocampus ingens
Siokunichthys nigrolineatus
Filicampus tigiris
Halicampus sp
Microphis ocellatus
Phycodurus eques
Choeroichthys sculptus
Entelerus aequoreus
Mitotichthys semistriatus
Stigmatopora nigra
Acentronura tentaculata
Doryrhamphus jansii
Nerophis spp
Hypselognathus rostratus
Hippicthys penicillus
Syngnathus louisianae
Stigmatopora argus
Dunckerocampus chapmani
Festucalex cinctus
Aulostomus chinensis
Hippocampus trimaculatus
Hippocampus zosterae
Vanacampus phillipi
Doryrhamphus excises
Corythoichthys intestinalis
Micrognathus andersonii
Solegnathus spinosissimus
Trachyrhamphus bioarctatus
Solegnathus hardwicki
Hippocampus abdominalis
Histiogamphelus briggsi
Pugnaso curtirostris
Solenostomus cyanopterus
Stigmatopora narinosa
Haliichthys taeniophorus
Hippocampus whitei
Kaupus costatus
Vanacampus margaritifer
Syngnathoides biaculeatus
Phyllopteryx taeniolatus
Hippocampus bargibanti
Urocampus carinirostris
Idiotropiscis lumnitzeri
Microphis doecata
Syngnathus exilisPseudophallus brasiliensis
Aulostomus maculatus
Vanacampus vercoi
Choeroichthys brachysoma
Leptoichthys fistularis
Doryrhamphus dactyliophorus
0.66
1
1
1
1
1
0.85
1
0.65
1
0.95
1
1
1
1
0.72
1
1
1
1
1
0.79
0.71
1
1
1
0.52
1
1
1
1
1
1
1
1
0.65
1
1
0.51
1
1
1
1
1
1
1
1
0.99
1
1
1
1
1
1
1
10.91
0.99
1
1
1.1
Stigmatopora nigra
Fistularia petimba
Choeroichthys sculptus
Siokunichthys nigrolineatus
Syngnathus fuscus
Microphis doecata
Doryrhamphus dactyliophorus
Nerophis sppLeptoichthys fistularis
Pseudophallus brasiliensis
Hippocampus zosterae
Solenostomus cyanopterus
Entelerus aequoreus
Hippocampus abdominalis
Maroubra perserrata
Choeroichthys brachysoma
Hippocampus pontohi
Doryichthys boaja
Hippocampus whitei
Stigmatopora narinosa
Cosmocampus elucens
Corythoichthys intestinalis
Hippicthys penicillus
Hippocampus bargibanti
Corythoichthys amplexus
Urocampus carinirostris
Microphis ocellatus
Hippocampus denise
Doryrhamphus jansii
Halicampus sp
Doryrhamphus excises
Syngnathus louisianae
Aulostomus maculatus
Hippocampus trimaculatusHippocampus ingens
Syngnathus exilis
Aulostomus chinensis
Dunckerocampus chapmani
Micrognathus andersonii
Hippocampus fuscus
Stigmatopora argus
Syngnathoides biaculeatusSolegnathus spinosissimus
Phycodurus equesPhyllopteryx taeniolatusSolegnathus hardwicki
Stipecampus cristatus
Trachyrhamphus bioarctatusHaliichthys taeniophorus
Filicampus tigirisIdiotropiscis lumnitzeriAcentronura tentaculata
Vanacampus margaritifer
Mitotichthys semistriatus
Vanacampus vercoi
Festucalex cinctus
Kaupus costatus
Pugnaso curtirostris
Histiogamphelus cristatusHistiogamphelus briggsi
Vanacampus poecilolaemusVanacampus phillipi
Hypselognathus rostratus
100
100
100
100
100
100
100
100
100
100
100100
100
100
100
100
100100
100
100
100
98
67
90
96
88
96
95
100
96100
98
80
67
85
99
79
9968
90
98
90
ResultsBayesian consensus tree Maximum parsimony strict consensus
L=20652
CI=0.228
RI=0.499
0.2
Corythoichthys amplexus
Maroubra perserrata
Hippocampus fuscus
Stipecampus cristatus
Hippocampus denise
Cosmocampus elucens
Doryichthys boaja
Syngnathus fuscus
Histiogamphelus cristatus
Hippocampus pontohi
Vanacampus poecilolaemus
Fistularia petimba
Hippocampus ingens
Siokunichthys nigrolineatus
Filicampus tigiris
Halicampus sp
Microphis ocellatus
Phycodurus eques
Choeroichthys sculptus
Entelerus aequoreus
Mitotichthys semistriatus
Stigmatopora nigra
Acentronura tentaculata
Doryrhamphus jansii
Nerophis spp
Hypselognathus rostratus
Hippicthys penicillus
Syngnathus louisianae
Stigmatopora argus
Dunckerocampus chapmani
Festucalex cinctus
Aulostomus chinensis
Hippocampus trimaculatus
Hippocampus zosterae
Vanacampus phillipi
Doryrhamphus excises
Corythoichthys intestinalis
Micrognathus andersonii
Solegnathus spinosissimus
Trachyrhamphus bioarctatus
Solegnathus hardwicki
Hippocampus abdominalis
Histiogamphelus briggsi
Pugnaso curtirostris
Solenostomus cyanopterus
Stigmatopora narinosa
Haliichthys taeniophorus
Hippocampus whitei
Kaupus costatus
Vanacampus margaritifer
Syngnathoides biaculeatus
Phyllopteryx taeniolatus
Hippocampus bargibanti
Urocampus carinirostris
Idiotropiscis lumnitzeri
Microphis doecata
Syngnathus exilisPseudophallus brasiliensis
Aulostomus maculatus
Vanacampus vercoi
Choeroichthys brachysoma
Leptoichthys fistularis
Doryrhamphus dactyliophorus
0.66
1
1
1
1
1
0.85
1
0.65
1
0.95
1
1
1
1
0.72
1
1
1
1
1
0.79
0.71
1
1
1
0.52
1
1
1
1
1
1
1
1
0.65
1
1
0.51
1
1
1
1
1
1
1
1
0.99
1
1
1
1
1
1
1
10.91
0.99
1
1
Results: Bayesian consensus tree
Urophori; tail-brooding fish
Outgroups
anus
Brood pouch
Gastrophori; abdominal-brooding fish
anus
Brood pouch
Results: “Doryrhamphinae”: Flagtail pipefishes
Flagtail pipefishes
Mostly freshwater pipefishes
DAVID HARASTI
Dunkerocampus Doryrhamphus
Results “Solegnathinae”: Sea Dragon clade
100
Solegnathus
Prehensile tails
Sygnathoides
David Harasti
Haliichthys taeniophorus
“Ribboned” sea dragon or pipefish
Results: Who is sister to seahorses?
Prehensile tails
All photos
David Harasti
Haliichthys Filicampus Trachyramphus
Conclusions
• Our molecular phylogeny is in agreement with Wilson(2003). Much revisionary work is needed at multiple taxonomic levels.
• The division of syngnathids into sub-families Syngnathinae and Hippocampinae is not supported; Syngnathinae is paraphyletic
• Maximum Parsimony and Bayesian phylogenetic analyses reproduce similar sets of clades, but differ in resolution of their arrangement
• Multiple origins of characters such as prehensile tails, freshwater habit, and fin presence/absence
• The seahorse's sister group is still unresolved
Future research in syngnathid systematics
• Further mapping of morphological characters to elucidate their origins and patterns in syngnathids
• Broader intraspecific geographic sampling
• Additional nuclear markers and increased axon sampling (21 genera to go!)
These future steps will further advance a
sufficiently robust phylogeny to support
syngnathid taxonomy, systematics,
biogeography, and conservation.
THANK YOU!
David
Harasti