Evolutionary history of the butterflyfishes (f: Chaetodontidae) and the rise of coral feeding fishes

Abstract

Of the 5000 fish species on coral reefs, corals dominate the diet of just 41 species. Most (61%) belong to a single family, the butterflyfishes (Chaetodontidae). We examine the evolutionary origins of chaetodontid corallivory using a new molecular phylogeny incorporating all 11 genera. A 1759-bp sequence of nuclear (S7I1 and ETS2) and mitochondrial (cytochrome b) data yielded a fully resolved tree with strong support for all major nodes. A chronogram, constructed using Bayesian inference with multiple parametric priors, and recent ecological data reveal that corallivory has arisen at least five times over a period of 12 Ma, from 15.7 to 3 Ma. A move onto coral reefs in the Miocene foreshadowed rapid cladogenesis within Chaetodon and the origins of corallivory, coinciding with a global reorganization of coral reefs and the expansion of fast-growing corals. This historical association underpins the sensitivity of specific butterflyfish clades to global coral decline.

Full text

Evolutionary history of the butterflyfishes (f: Chaetodontidae)
and the rise of coral feeding fishes
D. R. BELLWOOD*,S.KLANTEN*à,P.F.COWMAN*,M.S.PRATCHETT,N.KONOW*§
&L.VAN HERWERDEN*à
*School of Marine and Tropical Biology, James Cook University, Townsville, Qld, Australia
Australian Research Council Centre of Excellence for Coral Reef Studies, James Cook University, Townsville, Qld, Australia
àMolecular Evolution and Ecology Laboratory, James Cook University, Townsville, Qld, Australia
§Ecology and Evolutionary Biology, Brown University, Providence, RI, USA
Introduction
Coral reef fishes are a highly diverse group, with an
evolutionary history extending back more than 50 Myr
(Bellwood & Wainwright, 2002). From the fossil record,
it appears that scleractinian-dominated coral reefs and
modern coral reef fish families first appeared and then
diversified at approximately the same time, in the early
Cenozoic (Bellwood, 1996; Bellwood & Wainwright,
2002; Wallace & Rosen, 2006). This suggests that the
origins of modern coral reefs and their associated fish
families may be closely linked. However, it is remarkable
that of the 5000 or more fish species recorded from coral
reefs today only 128 eat corals (Cole et al., 2008; Rotjan &
Lewis, 2008) and just 41 are believed to feed directly on
scleractinian corals as their primary source of nutrition.
Moreover, 61% (25 of 41) belong to a single family, the
butterflyfishes (f. Chaetodontidae); of the remainder
most (eight) are in the Labridae. Why so few species
have been able to exploit such a widespread resource
remains a mystery. It also highlights the exceptional
abilities of the few corallivores that have managed to
subsist on corals, and the remarkable status of butter-
flyfishes. Despite being one of the most intensively
studied families of reef fishes, the evolution of this highly
specialized feeding mode remains poorly understood. In
particular, how many times has corallivory arisen within
the group and when did this unusual feeding mode first
arise? Did corallivory arise along with major coral groups
in the early Eocene?
Butterflyfishes are conspicuous and iconic inhabitants
of coral reef environments. The family contains over 130
species with representatives in all coral reef regions
(Allen et al., 1998; Kuiter, 2002). Their colourful pat-
terns, and ease of identification and observation have
ensured that the behavioural, ecological, morphological
and biogeographic characteristics of butterflyfishes have
been extensively studied (e.g. Motta, 1988; Ferry-
Graham et al., 2001a; Findley & Findley, 2001; Pratchett,
Correspondence: David R. Bellwood, School of Marine and
Tropical Biology, James Cook University, Townsville, Qld 4811, Australia.
Tel.: +61 7 4781 4447, fax: +61 7 4725 1570;
e-mail: david.bellwood@jcu.edu.au
ª2009 THE AUTHORS. J. EVOL. BIOL.
JOURNAL COMPILATION ª2009 EUROPEAN SOCIETY FOR EVOLUTIONARY BIOLOGY 1
Keywords:
biogeography;
chronogram;
coral reef;
innovation;
molecular phylogeny;
trophic novelty.
Abstract
Of the 5000 fish species on coral reefs, corals dominate the diet of just 41
species. Most (61%) belong to a single family, the butterflyfishes (Chae-
todontidae). We examine the evolutionary origins of chaetodontid corallivory
using a new molecular phylogeny incorporating all 11 genera. A 1759-bp
sequence of nuclear (S7I1 and ETS2) and mitochondrial (cytochrome b) data
yielded a fully resolved tree with strong support for all major nodes. A
chronogram, constructed using Bayesian inference with multiple parametric
priors, and recent ecological data reveal that corallivory has arisen at least five
times over a period of 12 Ma, from 15.7 to 3 Ma. A move onto coral reefs in
the Miocene foreshadowed rapid cladogenesis within Chaetodon and the origins
of corallivory, coinciding with a global reorganization of coral reefs and the
expansion of fast-growing corals. This historical association underpins the
sensitivity of specific butterflyfish clades to global coral decline.
doi:10.1111/j.1420-9101.2009.01904.x

2005). Indeed, they have been regularly identified as
indicator species of reef health (Reese, 1975; Roberts
et al., 1988; Kulbicki et al., 2005). It is this close associ-
ation with corals and coral reefs that stands as one of the
most important features of this family. With almost a
quarter of the species feeding on corals, they have what is
arguably the closest association of any fish group with
coral reefs. The key to understanding the history of this
relationship, however, is to obtain well-supported phy-
logenies based on multiple genes, and to use robust
molecular dating methodologies, informed by reliable
fossil data, to provide a temporal framework in which to
interpret recent ecological evidence.
Fossil evidence of increasing reef–fish interactions
points to a major change between the late Mesozoic
and the beginning of the Tertiary (Bellwood, 2003).
Although there is a diverse range of acanthomorph fishes
in the late Cretaceous, the earliest evidence of the vast
majority of extant reef fish families, for which there is a
fossil record, is from the Eocene. Most of these families
are first recorded from the 50-Myr-old deposits of Monte
Bolca, in northern Italy (e.g. Blot, 1980; Bellwood, 1996;
although it should be noted that a few molecular studies
have suggested that some lineages may predate the
Cretaceous ⁄Tertiary (K ⁄T) boundary, e.g. Streelman
et al., 2002; Alfaro et al., 2007; Azuma et al., 2008). The
Monte Bolca deposits mark the first modern coral reef
fish assemblage in terms of both its taxonomic compo-
sition and the functional attributes of the component
taxa (Bellwood, 1996), and it is here that we observe the
first evidence of increased interactions between fishes
and the benthos, with the appearance of several lineages
of fishes that were almost certainly grazing herbivores
(Bellwood, 2003). Although there have been many
Eocene fossils ascribed to the Chaetodontidae, a recent
evaluation of this material has rejected all of these taxa;
there is no reliable record of the family from the Eocene
(Bannikov, 2004). The oldest reliable fossil evidence for
the Chaetodontidae is of Miocene age (Carnevale, 2006).
However, with a robust molecular phylogeny we can
build on the fossil record and place recent ecological
advances in an evolutionary framework.
Recent ecological research has provided a new per-
spective on the nature of corallivory in reef fishes,
especially butterflyfishes. Building on existing informa-
tion (e.g. Birkeland & Neudecker, 1981; Blum, 1988;
Motta, 1988) recent studies have provided a detailed
understanding of the family in terms of feeding mor-
phology and kinematics (Ferry-Graham et al., 2001a,b;
Konow et al., 2008; Konow & Ferry-Graham, in press),
feeding strategies and behavioural interactions (Zekeria
et al., 2002; Gregson et al., 2008) and, most importantly,
the nature and extent of corallivory (Pratchett et al.,
2004; Berumen et al., 2005; Pratchett, 2005, 2007; Cole
et al., 2008; Rotjan & Lewis, 2008). It is now known that
butterflyfishes exhibit considerable diversity in the
nature of corallivory, underpinned by both ecomorpho-
logical and behavioural variation. This includes at least
two different modes of coral feeding, exploitation of both
soft and hard corals, and a spectrum of coral feeding
specializations ranging from highly specialized obligate
coral feeders that primarily target just one coral species to
facultative and generalist coral feeders that can feed on a
wide range of coral species (Pratchett et al., 2004;
Berumen et al., 2005).
The evolutionary history of these traits is poorly
understood. Existing morphological and molecular phy-
logenies have produced discordant tree topologies, open-
ing questions about the nature of character evolution.
Most molecular analyses of the family have focused on
relationships between species pairs or species complexes
(e.g. McMillan & Palumbi, 1995; Hsu et al., 2007) or have
included butterflyfishes as part of broader studies of
putative sister taxa (Bellwood et al., 2004). The most
comprehensive analysis to date using 3332 bp of mito-
chondrial and nuclear DNA yielded a well-supported
phylogeny, which differed significantly from all previous
topologies, and underpinned a thorough evaluation of
the systematics and taxonomy of the family (Fessler &
Westneat, 2007).
The evolutionary and ecological ramifications of these
relationships for corallivory, however, have yet to be
fully explored. We address four critical questions: (1)
How many times has corallivory arisen? (2) When did it
first arise? (3) Did corallivory and ⁄or a move onto coral
reefs underpin diversification within the family? and (4)
What are the broader implications for the evolution of
coral reefs? In this present study, we use a comprehen-
sive new molecular phylogeny to explore the evolution-
ary history of the family. For the first time, we use
molecular methods to examine the relationships between
all 10 genera and all respective subgenera (with the
exception of Roa Roa sensu, Blum, 1988; = Roa, Kuiter,
2002). We examined 56 species using two nuclear (ETS2
and S7I1) and a mitochondrial marker (cyt b); the nDNA
markers have not been used previously in this family. We
also apply, for the first time, current Bayesian analyses
for chronogram construction with multiple fossil calibra-
tions. Using this chronogram, ecological evidence and
diversification statistics, we explore the evolutionary
origins of corallivory and the timing of this highly
unusual fish–coral interaction. We then consider the
basis for these trophic innovations in the context of the
evolution of coral reefs.
Materials and methods
Taxon sampling
In total, 56 butterflyfish species were examined, with
multiple representatives of all 11 identified butterflyfish
genera and 12 subgenera (authorities given in Allen
et al., 1998). Specimens were obtained using spears
or nets with additional material obtained from the
2D. R. BELLWOOD ET AL.
ª2009 THE AUTHORS. J. EVOL. BIOL. doi:10.1111/j.1420-9101.2009.01904.x
JOURNAL COMPILATION ª2009 EUROPEAN SOCIETY FOR EVOLUTIONARY BIOLOGY

ornamental fish trade. A further eight species from the
Pomacanthidae, Ephippidae, Kyphosidae and Scatophag-
idae were included as putative outgroup species. Based on
Smith & Wheeler (2006), the Pomacanthidae then the
Ephippidae are the putative successive sister taxa to the
Chaetodontidae; the more distant kyphosid and micro-
canthid species were used to root the entire phylogeny.
Our taxon sampling specifically included 18 of the 26
species known to be obligate coral feeders (Appendix S1;
Table 1). Full taxon sampling in several lineages (Forcipiger,
Chelmon and Chelmonops) also enables us to explore the
temporal and biogeographic patterns of species origins.
Laboratory procedures
Total DNA was extracted from tissues using standard salt-
chloroform and proteinase K digestion extraction proce-
dures (Sambrook & Russell, 2001). Two nuclear genes,
ETS2 (ETS is a transcription factor important in cell
proliferation; Dwyer et al., 2007; Lyons et al., 1997); S7
Intron 1 (S7 is a ribosomal protein required for assem-
bling 16s RNA; Maguire & Zimmermann, 2001; Chow &
Hazama, 1998) and the mitochondrial protein-coding
region, cytochrome b(which participates in electron
transport; Kocher et al., 1989; Irwin et al., 1991;
McMillan & Palumbi, 1995) were used to explore the
evolutionary relationships among the butterflyfishes
(Appendix S1; Table 2). An average of two specimens
were sequenced for each species. Each 20 lL polymerase
chain reaction (PCR) volume contained 2.5 m
MM
Tris–Cl
(pH 8.7), 5 m
MM
KCl(NH
4
)
2
SO
4
, 200 l
MM
each dNTP,
MgCl
2
ranging from 1.5 to 4 m
MM
,10l
MM
each primer,
1 U of Taq Polymerase (Qiagen, Doncaster, Victoria,
Australia) and 10 ng template DNA. Amplifications
followed the same basic cycling protocol: an initial
denaturing step of 2 min at 94 C, followed by 35 cycles,
with the first five cycles at 94 C for 30 s, 30 s at primer-
specific annealing temperatures (T
a
) (Appendix S1; Table
2), followed by 1 min 30 s extensions at 72 C and the
remaining 30 cycles were performed as before, but at T
a
)2C. PCR products were purified by isopropanol pre-
cipitation (cyt band S7I1) or gel-purification on 2%
agarose gels, as two bands appeared routinely (ETS2).
This was also the case for S7I1 amplified fragments of
some species. A 500-bp fragment was retained for ETS2
whilst a 700-bp fragment was retained for S7I1.
Gel-excised fragments were purified in a column follow-
ing manufacturer’s protocols (Qiagen). Purified tem-
plates were quantified by UV-Vis absorbance (ND-1000
Spectrophotometer, NanoDrop

, Wilmington, NC, USA)
and sent to Macrogen Inc. (Seoul, South Korea) for direct
sequencing in both directions.
Analytical procedures
Data compilation
The consensus sequence of the multiple specimens
sequenced was used to represent each taxon. Sequences
were edited using Sequencher 4.5 (Gene codes corpora-
tion, Ann Arbor, MI, USA), and automatically aligned
using C
LUSTALLUSTAL
X (Thompson et al., 1997) and finally
manually corrected using S
E-E-
A
LL
version 2.0 available at
http://evolve.zoo.ox.ac.uk (Rambaut, 1996). Sequences
of this study are available at GenBank accession numbers
(ETS2: FJ167730–FJ167792; S7I1: FJ167793–FJ167846,
FJ167848–FJ167856 and cyt b: FJ167682–FJ167709,
FJ167711–FJ167719, FJ167721–FJ167729). Several
sequences of cytochrome bwere used from GenBank
(Chaetodon:C. argentatus AF108580, C. citrinellus AF108585,
C. kleinii AF108591, C. lineolatus AF108593, C. lunula
Table 1 Departure of chaetodontid lineages from global diversification rate estimated of the family Chaetodontidae.
Clade name Age Total ’=0 ’= 0.3 ’= 0.5 ’= 0.6 ’= 0.8 ’= 0.9
Crown group (CT) 32.8 130(56) r
G
= 0.1274 r
G
= 0.1246 r
G
= 0.1188 r
G
= 0.1141 r
G
= 0.0971 r
G
= 0.0787
CF 26.1 27(16) 7.61E-01 6.81E-01 6.61E-01 6.60E-01 6.76E-01 7.00E-01
CP 23.9 103(40) 4.23E-02 7.27E-02 9.32E-02 1.05E-01 1.42E-01 1.81E-01
AC 22.9 12(8) 8.84E-01 7.95E-01 7.76E-01 7.75E-01 7.94E-01 8.18E-01
C1* 17.8 3(1) 8.04E-01 8.49E-01 8.76E-01 8.89E-01 9.19E-01 9.38E-01
CH 17.8 93(38) 5.03E-04*2.89E-03*7.39E-03 1.16E-02 3.13E-02 6.29E-02
FH 14.2 16(8) 2.69E-01 3.01E-01 3.29E-01 3.50E-01 4.31E-01 5.19E-01
C3 13.4 21(13) 9.94E-02 1.44E-01 1.80E-01 2.04E-01 2.88E-01 3.83E-01
C2 12.6 37(14) 3.15E-03 1.14E-02 2.39E-02 3.46E-02 8.15E-02 1.49E-01
C4 11.3 31(11) 3.10E-03 1.12E-02 2.39E-02 3.50E-02 8.43E-02 1.57E-01
Ho 7.8 8(3) 2.01E-01 2.46E-01 2.81E-01 3.05E-01 3.96E-01 4.99E-01
PR 5.5 10(2) 2.07E-02 4.37E-02 7.08E-02 9.13E-02 1.69E-01 2.67E-01
C2 + C3 + C4 16.6 89(38) 1.61E-04*1.26E-03*3.90E-03 6.71E-03 2.21E-02 5.00E-02
C3 + C4 15.7 52(24) 5.44E-03 1.69E-02 3.14E-02 4.28E-02 8.81E-02 1.49E-01
Bold P-values highlight significantly higher species richness in subtending clade than expected under the global rate of cladogenesis
(*significance after Bonferroni correction). ’is the extinction rate, r
G
is the estimated global Chaetodontidae speciation rate conditional
on the extinction rate. Clade names and ages are taken from the node labels and mean node heights of Fig 1, and table 1 in Appendix S2.
Origins of coral feeding 3
ª2009 THE AUTHORS. J. EVOL. BIOL. doi:10.1111/j.1420-9101.2009.01904.x
JOURNAL COMPILATION ª2009 EUROPEAN SOCIETY FOR EVOLUTIONARY BIOLOGY

AF108594, C. meyeri AF108597, C. milliaris U23606,
C. multicinctus U23588, C. ornatissimus AF108600, C. plebius
AF108602, C. quadrimaculatus AJ748302, C. unimaculatus
AJ748304; Chelmon rostratus AF108612, Coradion altivelis
AF108613, Coradion chrysozonus AF108614, Forcipiger fla-
vissimus AF108615, Hemitaurichthys polylepis AF108616,
Heniochus acuminatus AF108618 and Parachaetodon ocellatus
AF108622 as per Littlewood et al., 2004; McMillan &
Palumbi, 1995; Nelson et al., unpublished GenBank
submission). Each locus was first examined for saturation
using DAMBE version 5.010 (Xia & Xie, 2001). This
method employs an entropy-based index of substitution
saturation (Iss); if Iss is significantly larger than the critical
Iss (i.e. Iss.c), sequences have experienced substitution
saturation (Xia et al., 2003). Prior to concatenating gene
regions, each gene was partitioned based on its function or
structure. Coding genes [cyt band a short exon region
(99 bp) of ETS2] were partitioned according to codon
positions (1st and 2nd combined as conserved region, and
3rd codon separately). Nuclear introns (ETS2 and S7I1)
were partitioned into putative stem (conserved) and loop
(hypervariable) regions. Eight separate gene partitions
were identified in total.
Phylogenetic analyses
Maximum parsimony (MP) analyses were implemented in
PAUPPAUP
* 4.0b10 (Swofford, 1998) using heuristic search
methods with 1000 pseudo-replicate bootstraps, tree-
bisection-reconnection branch swapping and random
addition of taxa. Two separate heuristic MP runs were
performed. First, all sites were treated equally and second,
sites were weighted according to gene partitions [3rd
codon, loop regions = 1; conserved (1st and 2nd codon)
and stem regions = 2]. A 50% majority rule consensus tree
was generated from all shortest trees obtained. Bayesian
inference (BI) analyses were implemented in MrBayes
version 3.1.2 (Huelsenbeck & Ronquist, 2001) using James
Cook University’s HPC GridSphere system (https://
ngportal.hpc.jcu.edu.au/gridsphere/). The analysis of the
combined data used a partition mix model method (pMM)
according to gene partitions with locus-specific substitu-
tion models, using M
RR
M
ODELTESTODELTEST
version 2.2 (Nylander,
2004) and Akaike information criterion (AIC) (Nylander
et al., 2004). Two Bayesian pMM analyses were performed
using Markov chain Monte Carlo (MCMC) simulations
with four chains of 2 000 000 generations each, sampling
trees every 100 generations. Stationarity was reached after
10 000 generations and a 50% majority rule consensus
tree was computed using the best 16 000 post-burn-in
trees from each run. Six putative sister taxa were included
in the analyses, three pomacanthids (Pomacanthus annu-
laris,P.rhomboids and P.sexstriatus), an ephippid (Platax
orbicularis), and two scats (Scatophagus argus and Selonotoca
multifasciata) and, in addition, two distant outgroups, a
kyphosid (Kyphosus vaigiensis) and a microcanthid (Tilodon
sexfasciatum), which were used to root resulting trees. The
single best tree was selected for molecular dating.
Maximum likelihood (ML) analysis was performed
using Garli version 0.95 (Zwickl, 2006). Ten independent
runs were performed using the best substitution model
(as per AIC) for the combined data (not partitioned)
implemented with M
ODELTESTODELTEST
version 3.7 (Posada &
Crandall, 1998). The best trees from the individual runs
were compared with ensure they did not differ in
topology and that the ML search was not arriving in a
suboptimal area of tree space. In addition, a ML analysis
with 100 bootstrap replicates was preformed to show
support of individual clades in the tree.
Molecular dating
Age estimation of the chaetodontid lineages was per-
formed in the program
BEASTBEAST
v1.4.8 (Drummond &
Rambaut, 2007).
BEASTBEAST
implements BI and a MCMC
analysis to simultaneously estimate branch lengths,
topology, substitution model parameters and dates based
on fossil calibrations. It also does not assume substitution
rates are autocorrelated across lineages, allowing the user
to estimate rates independently from an uncorrelated
exponential distribution or lognormal distribution
(UCLD). Many empirical data sets have been shown
not to demonstrate autocorrelation of rates and times
(Drummond et al., 2006; Alfaro et al., 2007; Brown et al.,
2008). An initial ultrametric tree was constructed in r8s
1.71 (Sanderson, 2004) from the topology and branch
lengths of the best Bayesian tree recovered from phylo-
genetic analyses, using a penalized likelihood (PL)
method (Sanderson, 2002). This topology was used for
calibration purposes by using parametric priors imple-
mented in
BEASTBEAST
to make assumptions a priori based on
fossil and biogeographic data (see below).
The vast majority of reported fossil chaetodontids are
demonstrably erroneous (Bannikov, 2004), a pattern
common in many other reef groups (e.g. scarids, Bellwood
& Schultz, 1991; pomacentrids, Bellwood & Sorbini,
1996). Fossil selection is critical, and fossils were only
used if placed in a family based on reliable morphological
criteria. Fossil calibrations were therefore restricted to
fossils from two families recorded from the Eocene
(50 Ma) deposits of Monte Bolca: Eoplatax papilio
(Ephippidae) (Blot, 1969) and Eoscatophagus frontalis
(Scatophagidae) (Tyler & Sorbini, 1999). With both
putative ephippid and scatophagid fossils having a min-
imum age of 50 Ma, an exponential prior was placed on
the Platax node (PL) with a hard lower bound age of 50 Ma
and a 95% soft upper bound of 65 Ma. The exponential
prior reflects the decreasing probability of a lineage being
older than its oldest fossil (Yang & Rannala, 2006; Ho,
2007). The soft upper bound of 65 Ma representing the
transition of fish faunas at the K ⁄T boundary (following
Bellwood & Wainwright, 2002; Bellwood et al., 2004;
Fessler & Westneat, 2007) beyond which there is no fossil
record of modern reef fish families.
BEASTBEAST
MCMC runs of 10 ·10
6
generations were
performed assuming the UCLD model with eight
4D. R. BELLWOOD ET AL.
ª2009 THE AUTHORS. J. EVOL. BIOL. doi:10.1111/j.1420-9101.2009.01904.x
JOURNAL COMPILATION ª2009 EUROPEAN SOCIETY FOR EVOLUTIONARY BIOLOGY

unlinked data partitions and unlinked substitution mod-
els specified by M
RR
M
ODELTESTODELTEST
v2.2 (Nylander, 2004).
Ten independent analyses were run sampling every
500th generation. Resulting log files were examined
using Tracer v1.4 (Rambaut & Drummond, 2007) to
ensure all analyses were converging on the same area in
tree space. Tree files (approx. )10% burn-in) were then
combined using LogCombiner (Rambaut & Drummond,
2007) and compiled into a maximum clade credibility
chronogram to display mean node ages and highest
posterior density (HPD) intervals at 95% (upper and
lower) for each node.
Optimizing ecological traits
Two ecological traits (corallivory and habitat use) were
mapped to the best phylogenetic tree that was the basis
for dating diversification in the chaetodontids, using
Mesquite v 2.6 (Maddison & Maddison, 2007). Habitat
use traits were scored as 0 = not on reefs, 1 = rocky reefs
and 2 = coral reefs. Diet traits were scored as 0 = non-
corallivore, 1 = omnivores (that have < 1% coral in the
diet and feed on other invertebrates and ⁄or algae),
2 = facultative or occasional coral feeders [which include
some (1–80%) hard coral in the diet] and 3 = corallivores
(in which the diet is dominated by i.e. > 80% hard or soft
corals). Ecological character states were drawn from the
published literature (Appendix S1; Table 1). Within the
corallivores, species are further identified as obligate hard
or soft corallivores when they feed exclusively on a
specific coral type.
Diversification rates
All diversification statistics were preformed in
RR
version
2.7.2 (http://www.Rproject.org) (Ihaka & Gentleman,
1996) using functions written for GEIGER (Harmon
et al., 2008), LASER (Rabosky, 2006) and associated
packages. The constant rates (CR) test of Pybus &
Harvey (2000) was used to investigate the rates of
cladogenesis of the chaetodontid crown group. This
test estimated the gamma statistic of the
BEASTBEAST
gener-
ated chronogram. Significantly negative gamma values
(< )1.645, one-tailed test) indicate a decrease in the
rates of cladogenesis over time. This implies that
internal nodes of the tree are distributed closer to the
root than would be expected under a Yule (pure birth)
process. To account for incomplete taxon sampling
(which increases Type 1 error of the CR test; Pybus &
Harvey, 2000) a Markov chain CR (MCCR) test (Pybus
& Harvey, 2000) was used to compare the observed
gamma to that of the null distribution created from
10 000 randomly subsampled, simulated (full) topolo-
gies under a Yule process. The relative cladogenesis
statistic (Nee et al., 1992) was used to identify lineages
with significantly faster ⁄slower rate of cladogenesis.
These methods have previously been used to investi-
gate diversification rates in tetraodontiform lineages
(Alfaro et al., 2007).
Methods implemented in the tetraodontiform study
(Alfaro et al., 2007) were used to calculate the global
diversification rate (r
G
) of the chaetodontids across
extinction rates (’) in increments of 0.1 from 0 to 0.9
(see Magallon & Sanderson, 2001). Using functions in
GEIGER (based on the method of moments estimator of
Magallon & Sanderson, 2001) the probabilities of the
observed species richness in each of the major chaeto-
dontid lineages were calculated using crown group ages
and the global estimates of diversification rate (r
G
) for
each increment of extinction. In case of the C. robustus
lineage, no other reported taxa in this clade were included
in this study and thus the stem group age estimator was
used (equation 10a, Magallon & Sanderson, 2001; see
Alfaro et al., 2007) to calculate the above probability.
Results
Sequence variability
We examined 1759 bp of sequence of which approxi-
mately 50% was parsimony-informative. The two nucle-
ar markers, ETS2 and S7I1 had 647 and 655 bp
respectively, cytochrome bcontributed a further 426 bp
with (47%, 65% and 47% parsimony-informative sites
respectively). None of the individual gene regions were
saturated (Iss 0.43 < 0.8 Iss.c, Iss 0.3 < Iss.c 0.8 and Iss
0.3 < Iss.c 0.78 respectively), neither was the concate-
nated data (Iss 0.4 < 0.8 Iss.c).
Model selection
The gene-specific models (AIC) for each of the eight
gene partitions used for Bayesian analysis were as
follows: Cyt bconserved region (gene 1, 1st and 2nd
codons) required a GTR + G model (gamma shape
parameter = 0.2397) with substitution Nst = 6, for its
3rd codon region (gene 2) a GTR + I + G model
(invariable sites = 0.02, c= 3.9890) with substitution
Nst = 6. ETS2 coding region required for its conserved
(gene 3) region a K80 model with substitution Nst = 2
(ti ⁄tv ratio = 2.5419), and its variable (gene 4, 3rd
codon) region a HKY model with substitution Nst = 2
(ti ⁄tv ratio = 1.5633). Both ETS2 stem (gene 5) and
loop (gene 6) required a GTR + G model (c= 0.9426
and 1.2595 respectively) with Nst = 6 substitution clas-
ses. S7I1 stem (gene 7) region required a HKY + G
model (c= 1.5603) with substitution Nst = 2 (ti ⁄tv
ratio = 1.3058) and its loop regions (gene 8) a GTR + G
model (c= 5.4651) with substitution Nst = 6. The
model selections for pMM BI only requires a general
‘form’ of the model (Nylander, 2004), as the Markov
chain integrates uncertainties of the parameter values.
Therefore, seven of the eight gene partitions had a base
frequency = dirichlet (1,1,1,1) (i.e. unequal) while the
eight gene (gene 3, ETS2 coding, conserved region) base
frequency was set to = fixed (equal).
Origins of coral feeding 5
ª2009 THE AUTHORS. J. EVOL. BIOL. doi:10.1111/j.1420-9101.2009.01904.x
JOURNAL COMPILATION ª2009 EUROPEAN SOCIETY FOR EVOLUTIONARY BIOLOGY

Maximum likelihood analysis (GARLI) required an
overall GTR + G model (c= 0.5120) for all regions,
substitution Nst = 6 with substitution rates fixed
(0.7160, 2.3497, 0.8239, 0.6913 and 3.8464), and base
frequencies fixed (0.26, 0.2336, 0.2115 and 0.2949).
Tree inference
Stationarity of the Bayesian analyses was reached after
much fewer then 10 000 generations in both runs,
(visualized in Tracer version 1.4; Rambaut & Drum-
mond, 2007) and the 50% majority rule consensus tree
topology was no different from the best trees of each run
()lnL = )22 255.116 1st run and lnL = )22 254.149 2nd
run) with very high posterior probabilities (Fig. 1). Both
MP and ML analyses inferred the same tree topology as
per Bayesian analysis. We therefore included only the
support for each retrieved node (Fig. 1). Four major
clades of Chaetodon were retrieved, resembling closely the
four clades retrieved in a previous molecular study by
Fessler & Westneat (2007). Although identical species
were not analysed in the two studies, the placement of
species common to both studies was identical, despite the
use of different loci in the two studies. For clarity, in
Chaetodon we follow the four clades of Fessler & Westneat
(2007). Old taxonomic groupings were found to be of
limited utility (only one remains intact and retains its
traditional boundaries (Radophorus in clade 4) and we
will not consider them further within Chaetodon. Fessler
& Westneat (2007) provide a thorough evaluation of the
taxonomy of the family. The only additional detail from
our study is that Parachaetodon would make Chaetodon
(and the subgenus Discochaetodon) paraphyletic and
Parachaetodon is probably best placed within Chaetodon
(as a junior synonym).
Molecular dating
The best Bayesian topology and branch lengths received
by phylogenetic analyses (Fig. 1) was used as the initial
starting tree with an exponential prior used to calibrate
the PL node (see Materials and methods).
BEASTBEAST
log files
analysed in Tracer showed convergence between inde-
pendent runs in tree space. High effective sample size
scores of individual parameters indicated valid estimates
based on independent samples from the posterior distri-
bution of the MCMC. A maximum clade credibility
chronogram was compiled in Tree Annotator from
180 000 post-burn-in trees (9 ·10
7
generations from
10
BEASTBEAST
MCMC runs). The chronogram displays mean
node heights received at each node by
BEASTBEAST
MCMC
with bars representing 95% HPD (Appendix S2, Fig. 1).
The family Chaetodontidae dates back to the early
Eocene where it split from pomacanthids with a mean
age of the most recent common ancestor (MRCA) of
50.1 Ma (41.5–60.7, 95% HPD). Estimated ages indicate
the origin of the butterfly fish and bannerfish clades at a
mean age of 32.8 Ma (24.9–40.9, 95% HPD) after which
they rapidly diversified, with the four major Chaetodon
lineages in place by the mid Miocene (Appendix S2;
Table 1). Also during the early Miocene we see the
origins of the three major divisions within the bannerfish
clade.
Optimization of ecological traits
Based on the species examined it is clear that corallivory
has arisen on at least five separate occasions (Fig. 2).
Corallivory has been reported in 25 chaetodontid species
(Appendix S1; Table 1). All are in the reef-butterflyfishes
clade and are restricted to a single monophyletic genus,
Chaetodon. Of these 25 corallivores, 17 are included in the
current phylogeny. The remaining eight species are easily
included in the four main Chaetodon clades based on
previous phylogenetic and taxonomic evidence (Appen-
dix S1; Table 1).
Chaetodon clade 1 contains only three species, all are
restricted to West African coastal waters, with no record of
coral feeding. Chaetodon clade 2 (37 species) contains three
distinct lineages of coral feeders (estimated MRCA to
omnivorous sister taxa in parentheses): C.quadrimaculatus
(3.2 Ma), the C.multicinctus clade (4.9 Ma) and the
C.unimaculatus-interruptus clade (4.3 Ma). The first two
lineages are hard coral feeders and probably have an
obligate dependence on corals. Chaetodon unimaculatus and
C.interruptus feed on soft and hard corals. Chaetodon clade 3
is predominantly corallivorous, with 19 of the 21 species
being obligate corallivores. Of the remaining species, the
diet of C.tricinctus is unknown, leaving one noncorallivore,
Parachaetodon ocellatus. The chronogram places the origins
of corallivory at about 15.7 Ma. Of the 31 species in
Chaetodon clade 4 only two are corallivores: C.melannotus
and C.ocellicaudus. These sister species are both obligate
soft coral feeders (Appendix S1; Table 1). They separated
from their omnivorous sister at about 9.8 Ma.
Diversification rates
The relative cladogenesis statistics identified the Chaeto-
don lineage as having a significantly different rate of
cladogenesis than its sister lineage. Both CR and MCCR
tests showed no evidence for a slowdown in the rate of
cladogenesis through time for the family Chaetodontidae
(c=)1.248, MCCR adjusted P= 0.55). As noted by
Magallon & Sanderson (2001) the estimates of r
G
decreased with increasing extinction rates (Table 1).
The Chaetodon clade (CH) with all subtending lineages
showed significantly higher species diversity than
expected given the global diversification rate up to 90%
extinction rate (adjusted P= 0.03; ’= 0.9). Clades 2 and
4 (C2 and C4) showed significantly higher species
diversity than expected (up to 80% extinction), however,
the corallivorous clade 3 (C3) is not significantly more
diverse than expected given the crown diversification
6D. R. BELLWOOD ET AL.
ª2009 THE AUTHORS. J. EVOL. BIOL. doi:10.1111/j.1420-9101.2009.01904.x
JOURNAL COMPILATION ª2009 EUROPEAN SOCIETY FOR EVOLUTIONARY BIOLOGY

rate even in the absence of extinction. The Prognathodes
lineage also shows significantly higher diversification for
up to 30% extinction (P= 0.044). If using a Bonferroni
correction (adjusted P= 0.0038) the Chaetodon (CH and
C2 + 3 + 4) clade still remains significant at low extinc-
tion rates.
Fig. 1 Inferred phylogeny of the butterflyfish and bannerfish (f. Chaetodontidae), based on 56 species with representatives from all
11 genera and 12 subgenera, obtained by Bayesian, maximum parsimony (MP) and maximum likelihood analyses for three loci (ETS2, S7I1
and cyt b). The topology shows the best bayesian tree with posterior probabilities (consensus of 32 000 trees) and bootstrap support (> 50%)
of MP and ML (1000 and 100 bootstrap replicates respectively). (*) 100% support. (–) no bootstrap support. The tree was rooted with Kyphosus
vaigiensis and Tilodon sexfasciatus.
Origins of coral feeding 7
ª2009 THE AUTHORS. J. EVOL. BIOL. doi:10.1111/j.1420-9101.2009.01904.x
JOURNAL COMPILATION ª2009 EUROPEAN SOCIETY FOR EVOLUTIONARY BIOLOGY

Discussion
Systematics of the Chaetodontidae
We present a comprehensive evaluation of the Chae-
todontidae, with representatives from all described
genera and currently recognized subgenera. There was
an extremely high degree of congruence among gene
regions and among methods (Likelihood, Parsimony and
Bayesian). Using independent models for each gene
partition the resultant phylogeny had strong support for
all major nodes. In all analyses, the phylogeny strongly
Fig. 2 A chronogram of the Chaetodontidae with optimized trophic modes reveals five independent origins of corallivory over the
last 15.7–3.2 Ma. Red dotted branches indicate obligate hard coral feeders and blue dashed branches obligate soft coral feeders. The
estimated ages are in Ma (see Fig. 1 and table 1 in Appendix S2 and for confidence intervals of the mrca age estimates). The butterflyfish
illustrations exemplify some of the corallivores in each of the independent clades in which coralivory has arisen (images from Kuiter, 2002).
8D. R. BELLWOOD ET AL.
ª2009 THE AUTHORS. J. EVOL. BIOL. doi:10.1111/j.1420-9101.2009.01904.x
JOURNAL COMPILATION ª2009 EUROPEAN SOCIETY FOR EVOLUTIONARY BIOLOGY

supports the monophyly of the family with a basal split
into two clades: the long-snouted bannerfishes and the
reef butterflyfishes.
Our phylogeny identified three major divisions in the
long-snouted bannerfish clade and four divisions in
the reef butterflyfish clade. A comparable pattern, for
nine of the 10 genera, was reported by Fessler &
Westneat (2007). Our data support this earlier study in
placing Amphichaetodon within the bannerfish clade,
rather than as a sister taxon to all remaining species in
the family, as suggested by most morphological phy-
logenies (Smith et al., 2003). The degree of agreement
between the two molecular studies is noteworthy.
Despite using different markers and different represen-
tative species, the topology of the resultant trees were
almost identical. This provides excellent independent
corroboration of our tree. With a robust, well sup-
ported phylogenetic reconstruction for this family we
are now able to explore the evolutionary history of
corallivory.
Divergence times within the Chaetodontidae
Given a well-supported cladogram, with independent
support for the topology, we endeavoured to provide
robust molecular age estimates within a chronogram. The
use of the program
BEASTBEAST
allowed more precise age
calibrations than previous approaches. Furthermore, the
use of exponential priors accommodates both the influ-
ence of the faunal transition at the K ⁄T boundary and
the stronger influence of the 50 Ma calibration based on
the fossil ephippid (Eoplatax). Our age estimates in the
resultant chronogram are supported by several indepen-
dent lines of evidence.
Firstly, our estimated ages agree well with the available
fossil record. We used the two best fossil dates for
calibration [i.e. (1) The K ⁄T boundary, marking the
transition between Mesozoic and Cenozoic faunas
(Patterson, 1993; Bellwood & Wainwright, 2002) and,
(2) the calibration 50 Ma, marking the earliest fossil
record of numerous reef fish families (Bellwood, 1996)].
However, there is a third piece of fossil evidence: a fully
articulated Miocene fossil chaetodontid (Carnevale,
2006). It is morphologically extremely similar to extant
taxa in clade 4, and at 7 Ma old lies shortly after the age
estimates for this clade with a mean age of 11.3 and 95%
HPD of 7.9–15.2 Ma.
Secondly, as an independent check, we can compare
our estimated ages with major biogeographic events.
These again compare favourably. Firstly, the terminal
Tethyan event (TTE) marking the final closure of the
Red Sea land bridge is dated between 12 and 18 Ma
(Steininger & Ro
¨gl, 1984). These ages approximate the
minimum age of the initial division between the major
clades within Chaetodon at 17.8 Ma (13.3–23.2 Ma, 95%
HPD). Of the major clades, two (2 and 4) lie on either side
of the land bridge, whereas clade 1 is restricted to the
Atlantic and clade 3 is restricted to the Indo-Pacific.
Secondly, the ages of lineages that appear to have been
separated by the rising of the Isthmus of Panama (IOP)
i.e. Chaetodon humeralis–C.ocellatus at 3.4 Ma (1.8–
5.4 Ma, 95% HPD) are again extremely close to the
estimated final closure of this land bridge at 3.1 Ma
(Coates et al., 1992), with the 95% density distribution
encompassing the geological dates (Lessios, 2008).
Finally, divisions between the Indian Ocean and Pacific
Ocean pairs (e.g. C.unimaculatus–interruptus at 1.3 Ma
and C.trifasciatus at 2.4 Ma) closely match the estimated
ages of other Indian Ocean–Pacific Ocean divisions
(McCafferty et al., 2002; Read et al., 2006). The separa-
tion of C.sedentarius and C. sanctaehelenae from their
closest known sister lineage in the Indo-Pacific may be a
further example of an invasion of the Atlantic via the
Cape of Good Hope (reviewed in Floeter et al., 2008).
Based on the first two biogeographic divisions (TTE and
IOP), our estimated ages with HPD intervals closely
approximate these two well dated biogeographic events.
Finally, a comparison of our age estimates with those
in previous studies, using a range of calibration methods,
suggest that the estimated ages of our terminal taxa are
comparable with those of other reef fishes (e.g. Fauvelot
et al., 2003; Bernardi et al., 2004; Klanten et al., 2004;
Barber & Bellwood, 2005; Read et al., 2006; Cowman
et al., 2009) and other reef organisms (Palumbi et al.,
1997; Lessios et al., 1999; Renema et al., 2008). The
closest study to the present work is by Fessler & Westneat
(2007) which yielded a very similar phylogeny and
broadly comparable ages, even though they used a single
model in tree construction and an additive PL method for
age estimation, while we used a partitioned mixed model
and Bayesian MCMC analyses with informative prior
calibrations. These differences will not necessarily change
the tree topology but can change relative branch lengths,
while the
BEASTBEAST
analyses take into account uncertainty
in topology, sequence dataset and model parameters.
Overall, fossil, biogeographic and comparative data pro-
vide strong support for our chronogram. This provides a
relatively robust platform for evaluating the evolution of
corallivory on coral reefs.
Evolutionary and biogeographic patterns within the
Chaetodontidae
In the Chaetodontidae, a move onto reefs was associ-
ated with a significant increase in species richness.
Interestingly, there was no increase associated with a
switch to corallivory and the exploitation of this widely
available reef resource. The Chaetodontidae can be
effectively divided into two ecologically and morpho-
logically distinct clades that should be represented as
sub-families: the bannerfishes and the butterflyfishes.
The bannerfish clade is characterized by a distinctive
long-snout morphology and it is within this clade that
we see a novel suspensorial protrusion mechanism
Origins of coral feeding 9
ª2009 THE AUTHORS. J. EVOL. BIOL. doi:10.1111/j.1420-9101.2009.01904.x
JOURNAL COMPILATION ª2009 EUROPEAN SOCIETY FOR EVOLUTIONARY BIOLOGY

(Ferry-Graham et al., 2001a). Despite the morphological
variation and innovation within the bannerfish clade,
however, the standing species richness of the bannerfish
lineage is not significantly different than expected given
the global rate of cladogenesis (even at high extinction
rates) for the crown Chaetodontidae (Table 1). Biogeo-
graphically, the bannerfish clade has close links with
Australia and temperate or sub-tropical waters, and a
subtropical Australian origin for this clade remains a
distinct possibility. Although habitat optimization in the
bannerfishes is uncertain (Fig. S1), three of the eight
lineages are found on temperate subtropical rocky reefs
and many species in the other lineages are found in
rocky or coastal waters, supporting these temperate
associations.
In contrast to the bannerfishes, the butterflyfishes
exhibit limited morphological variation. Indeed, they
appear to be relatively uniform (Motta, 1988) with
relatively simple oral jaw mechanics and kinematics
(Ferry-Graham et al., 2001a). Only with respect to
intramandibular flexion does there appear to be any
clear morphological variation (Konow et al., 2008).
Depending on the definition of a coral reef and a coral
reef fish (cf. Bellwood & Wainwright, 2002), it appears
that there have been multiple invasions of coral reefs
by chaetodontids. The butterflyfish clade contains
103 species, approximately 80% of species within
the family, and is strongly associated with coral reefs.
As in parrotfishes (Streelman et al., 2002), wrasses
(Westneat & Alfaro, 2005) and tetraodontoids (Alfaro
et al., 2007), the reef dwelling clades are exceptionally
species rich. The Chaetodon clade, in particular, exhibits
far higher numbers of species than expected even at
high extinction rates (P= 0.03, ’=0.8). When consider-
ing just the reef-based clades 2, 3 and 4 there is a
greater significant difference (P= 0.021, ’= 0.8) from
expected. It thus appears that a move to reefs did indeed
underpin diversification in Chaetodon, as previously
reported in the tetraodontiformes (Alfaro et al., 2007).
This pattern may be expected in a number of reef fish
groups (Bellwood & Wainwright, 2002). However, it is
noteworthy that clade 3 does not demonstrate higher
species richness than expected, even though this obli-
gate reef fish clade contains the largest number of
corallivores found in any teleost taxon. It appears that a
move onto reefs, not a switch to corallivory, under-
pinned diversification within the family.
The rise of corallivory
The Chaetodontidae contains more corallivores than any
other fish family; however, this did not arise as a result of
a single exceptional event. Corallivory has arisen at least
5 times, with representatives in almost every major
butterflyfish clade. Furthermore, it appears to have
arisen relatively recently (15.7–3.2 Ma) and in a number
of markedly different ways.
The oldest estimated record of corallivory is in Chaeto-
don clade 3 at 15.7 Ma (the MRCA with an omnivorous
sister lineage; Fig. S2). Of the 13 species examined in this
clade, 12 are corallivorous (the exception is Parachaetodon
ocellatus). Nine additional species can be placed in this
clade based on phylogenetic (austriacus,larvatus,octofas-
ciatus,speculum and zanzibarensis) and taxonomic (mel-
apterus,lunulatus,andamanensis and triangulum) evidence
(Fessler & Westneat, 2007; Hsu et al., 2007). All these
taxa are obligate corallivores. This is the oldest record of
corallivory in the family and it is in clade 3 that we see
the strongest reef associations and the tightest links
between fishes and corals. Several species feed on just
one or two coral species and may be incapable of
switching prey species (Berumen & Pratchett, 2008),
while others specialize by ingesting specific parts of the
coral or just mucous (Cole et al., 2008). These species
have relatively long intestines and appear to represent an
extreme level of coral feeding specialization (Elliott &
Bellwood, 2003; Konow & Ferry-Graham, in press).
Given this level of specialization, it is no surprise that it is
species within this clade that exhibit the most extreme
negative response to the decline in coral cover as a result
of anthropogenic disturbances and climate change
(Pratchett et al., 2006, 2008; Wilson et al., 2006).
Given this long association with corallivory, the
monotypic Parachaetodon was a striking inclusion in clade
3. Parachaetodon ocellatus is not a corallivore and often
lives in sheltered sediment rich areas (Allen et al., 1998).
Given its position in the tree, this appears to be the first
recorded reversal from corallivory to omnivory. The
evolutionary scenario that may have triggered such a
change is unclear. The explanation may be biogeograph-
ic, with a dietary switch following the loss of corals in an
isolated marine basin.
The second oldest record of corallivory is in Chaetodon
clade 4 at about 9.8 Ma, in C.melannotus and its sister
species C.ocellicaudus (cf. Fessler & Westneat, 2007; Hsu
et al., 2007). These species are again strongly reef asso-
ciated and highly specialized obligate coral feeders.
However, these taxa are restricted exclusively to soft
corals. Their relationship with other members of the
clade is not well resolved and a sister group relationship
with the omnivore C.selene suggested by Fessler &
Westneat (2007) would imply that the origins of coral-
livory in the melannotus–ocellicaudus clade are younger
than our estimate. Nevertheless, this represents an
independent, and highly distinctive, obligate soft coral
feeding lineage.
The most recent examples of corallivory are found in
Chaetodon clade 2. This clade contains a large number of
species that occasionally graze on live corals, but only
four obligate corallivores. Here, corallivory arose as a
result of three independent events: C.multicinctus clade
(inc. pelewensis and punctatofasciatus) at about 4.9 Ma,
C.quadrimaculatus at about 3.2 Ma and the C.unimacul-
atus–interruptus clade at about 4.3 Ma. These ages are not
10 D. R. BELLWOOD ET AL.
ª2009 THE AUTHORS. J. EVOL. BIOL. doi:10.1111/j.1420-9101.2009.01904.x
JOURNAL COMPILATION ª2009 EUROPEAN SOCIETY FOR EVOLUTIONARY BIOLOGY

well-established as incomplete taxon sampling precludes
robust estimates. Nevertheless, all the three stand as
relatively recent independent events, a pattern that is
unlikely to be altered by further taxon sampling. There
are two different feeding modes. The first two lineages
contain obligate corallivores and in both cases the pre-
ferred coral prey appear to be Pocillopora spp. (Berumen &
Pratchett, 2006). The latter clade consists of two sister
taxa that feed exclusively on corals; C.unimaculatus in the
Pacific and C.interruptus in the Indian Ocean. Chaetodon
unimaculatus appears to be unique in that it feeds on both
soft and hard corals (hard in French Polynesia and
Hawaii vs. soft on the GBR and in Guam; Motta, 1988;
Wylie & Paul, 1989; Konow & Ferry-Graham, in press;
M.S. Pratchett, unpublished). It is also the only butterf-
lyfish to take large bites from corals that remove both the
polyp and the surrounding tissues. In this, the bite is
more reminiscent of excavating parrotfishes which leave
distinctive scars at the feeding site (Bellwood & Choat,
1990). This robust feeding mode is reflected by an
unusually robust jaw morphology in this lineage (Motta,
1988; Konow et al., 2008).
Despite the clear patterns, care is needed when
interpreting evolutionary history from phylogenies.
The ages of origination refer to the approximate ages
at which extant lineages are hypothesized to have
commenced corallivory. The ages of these taxa are
comparable with those recorded from other reef fish
families such as the Pomacentridae (e.g. McCafferty
et al., 2002), Labridae (e.g. Read et al., 2006; Cowman
et al., 2009) and Acanthuridae (Klanten et al., 2004). Yet
in each of these three families the Eocene fossil record
yields several extinct fossil taxa that are the functional
equivalents of extant taxa (Bannikov & Sorbini, 1990;
Bellwood & Sorbini, 1996; Tyler & Sorbini, 1999). One
can not, therefore, discount the possibility that coralli-
vory predated the origins of extant lineages and our
minimum age estimates. However, our estimates do
provide a clear indication of the minimum age of this
feeding mode and evidence of an increasing diversity of
corallivores, in terms of both feeding modes and
number of lineages, during the Miocene and Pliocene
(15.7–3 Ma).
Corallivory and its implications for reef–fish
interactions and the evolution of coral reefs
Our chronogram clearly suggests that corallivory did not
arise with the origins of the major coral groups in the
Eocene. Rather, it ties in with a major expansion and
reorganization of reefs in the Miocene, and coincides
with the initial formation of the biodiversity hotspot in
the Indo-Australian Archepilago.
Even given that our estimates are minimum ages, 15.7–
3 Ma still represents a relatively recent origination for
such a derived feeding mode as corallivory. Scleractinian
corals have been a significant component of shallow
carbonate reefs since the early Tertiary, with most of the
major Acropora clades (the coral genus targeted by most
modern corallivores) already represented in the Eocene at
49–37 Ma (Wallace & Rosen, 2006). In contrast, other
major coral reef benthic feeding modes, e.g. grazing
herbivory and crushing with pharyngeal jaws, have been
present for at least 50 Ma (Bellwood & Sorbini, 1996;
Bellwood, 1999, 2003; Cowman et al., 2009). The prob-
lem of minimum estimates notwithstanding, this rela-
tively recent rise of corallivory raises two questions: are
chaetodontids one of the most recent taxa to switch to
corallivory and does this switch reflect a broader change
in the nature of reef–fish interactions?
In terms of the evolution of corallivory, the evidence is
scarce but all the indications are that the timing of
corallivory in chaetodontids is comparable to that of the
only other major group with significant numbers of
corallivores, the labrids. Based on the most recent labrid
phylogeny (Cowman et al., 2009) corallivory appears to
be derived, to have arisen only once (in the Labropsis–
Labrichthys clade) and to have arisen relatively recently,
although considerably earlier than in the Chaetodontidae
(at 29 Ma). In the parrotfishes (i.e. Bolbometopon muric-
atum and Sparisoma viride) coral feeding probably arose
prior to the late Miocene (12 and 10 Ma respectively;
Robertson et al., 2006; Cowman et al., 2009). Overall, it
appears that the chaetodontids are only exceptional in
terms of the number of corallivorous species within the
family. Their dietary shift appears to have coincided with
a general rise in corallivory in a range of reef fish families.
In terms of the broader changes in the nature of reef–
fish interactions, the rise of corallivory in the Miocene is
consistent with several other lines of evidence. We see
a progressive increase in detritivory in the Miocene
(Harmelin-Vivien, 2002) and a number of novel special-
ist groups e.g. specialist foraminifera feeders and fish
cleaners (Macropharyngodon; Read et al., 2006; Cowman
et al., 2009). The origins of corallivory, therefore, fit in a
broader context in which the Miocene exhibits a new
level of reef–fish interactions with more specialized reef-
associated taxa. This may be associated with the rise
of Acropora and Pocillopora as the dominant coral groups
during this period (Johnson et al., 2008; B. Rosen, personal
communication). The vast majority of corallivores and all
obligate specialists feed only on these coral genera.
The number of independent origins of corallivory and
the lack of morphological modifications to the feeding
apparatus suggest that there are few morphological
restrictions to corallivory, although the elongation of the
intestine suggests that the difficulty, if any, may lie in
processing rather than procuring coral tissues. Extant
corallivores are often highly selective feeders, exploiting
specific coral species or even specific sites on a coral (e.g.
damaged tissues) (McIlwain & Jones, 1997). The rise of
corallivory may therefore have been dependent on corals
reaching sufficient densities to permit the selective feeding
necessary to adequately process the coral tissues; the
Origins of coral feeding 11
ª2009 THE AUTHORS. J. EVOL. BIOL. doi:10.1111/j.1420-9101.2009.01904.x
JOURNAL COMPILATION ª2009 EUROPEAN SOCIETY FOR EVOLUTIONARY BIOLOGY

increased access to Acropora and Pocillopora colonies trig-
gering the expansion of corallivory in the Miocene. The
rapid expansion of coral bearing carbonate platforms in the
Indo-Australian Archepelago in the early-mid Miocene
(Wilson, 2008) may therefore have acted as the trigger for
not only the rapid expansion of numerous fish groups but
the origins of trophic novelty, including corallivory
(Renema et al., 2008; Cowman et al., 2009). In this
context, it is interesting to note that on modern coral reefs
the number of corallivores declines swiftly in response to
the loss of coral cover (Pratchett et al., 2006, 2008).
Coral reefs have been exposed to escalating predation
pressure for millennia (Vermeij, 1977; Bellwood, 2003).
For corals, predation by fishes certainly appears to have
increased over the last 15 Ma. We now have, for the first
time, an understanding of the origins of corallivory in
fishes. Of all corallivorous fishes 63% are found within a
single family, the Chaetodontidae. Yet, surprisingly,
within this family this derived feeding mode has arisen
at least five times over the last 3–15.7 Ma, with specialists
on both soft and hard corals. This unusual feeding mode
appears to reflect an exceptionally close association
between this family and coral reefs. An understanding
of this history offers a new perspective on the nature of
the relationships between fishes and coral reefs in a
changing world.
Acknowledgments
The authors wish to thank C. Fulton, A. Hoey, P.C.
Wainwright, F. Walsh and P. Wirtz for tissue samples; S.
Wismer for graphics assistance; the staff of Carrie Bow
Cay, Orpheus Island, Lizard Island and Moorea CRIOBE
Research Stations for invaluable field support, colleagues
in the Centre of Excellence for Coral Reef Studies for
helpful discussions and several reviewers for constructive
comments. This work was financially supported by James
Cook University and the Australian Research Council.
References
Alfaro, M.E., Brock, C.D. & Santini, F. 2007. Do reefs drive
diversification in marine teleosts? Evidence from the pufferf-
ishes and their allies (Tetraodontiformes: Acanthomorpha).
Evolution 61: 2104–2126.
Allen, G.R., Steene, R. & Allen, M. 1998. A Guide to Angelfishes
and Butterflyfishes. Odyssey Publishing-Tropical Reef Research,
Perth.
Azuma, Y., Kumazawa, Y., Miya, M., Mabuchi, K. & Nishida, M.
2008. Mitogenomic evaluation of the historical biogeography
of cichlids toward reliable dating of teleostean divergences.
BMC Evol. Biol. 8: 215.
Bannikov, A.F. 2004. Fishes from the Eocene of Bolca,
northern Italy, previously classified with the Chaetodontidae
(Perciformes). Studie Ricerche sui Giacimenti Terziari di Bolca
10: 55–74.
Bannikov, A.F. & Sorbini, L. 1990. Eocoris bloti, a new genus and
species of labrid fish (Perciformes, Labroidei) from the Eocene
of Monte Bolca, Italy. Studie Ricerche sui Giacimenti Terziari di
Bolca 6: 133–148.
Barber, P.H. & Bellwood, D.R. 2005. Biodiversity hotspots:
evolutionary origins of biodiversity in wrasses (Halichoeres:
Labridae) in the Indo-Pacific and New World tropics. Mol.
Phylogenet. Evol. 35: 235–253.
Bellwood, D.R. 1996. The Eocene fishes of Monte Bolca: the
earliest coral reef fish assemblage. Coral Reefs 15: 11–19.
Bellwood, D.R. 1999. Fossil Pharyngognath fishes from Monte
Bolca, Italy, with a description of a new pomacentrid genus
and species. Studi e Ricerche sui Giacimenti Terziari di Bolca 8:
207–217.
Bellwood, D.R. 2003. Origins and escalation of herbivory in
fishes: a functional perspective. Paleobiology 29: 71–83.
Bellwood, D.R. & Choat, J.H. 1990. A functional analysis of
grazing in parrotfishes (family Scaridae): the ecological impli-
cations. Environ. Biol. Fish 28: 189–214.
Bellwood, D.R. & Schultz, O. 1991. A review of the fossil record
of the parrotfishes (Labroidei: Scaridae) with a description of a
new Calotomus species from the Middle Miocene (Badenian) of
Austria. Ann. Nat. Mus. Wien. 92: 55–71.
Bellwood, D.R. & Sorbini, L. 1996. A review of the fossil record
of the Pomacentridae (Teleostei: Labroidei) with a description
of a new genus and species from the Eocene of Monte Bolca,
Italy. Zool. J. Linn. Soc. 117: 159–174.
Bellwood, D.R. & Wainwright, P.W. 2002. The history and
biogeography of fishes on coral reefs. In: Coral Reef Fishes:
Dynamics and Diversity in a Complex Ecosystem (P.F. Sale, ed.),
pp. 5–32. Academic Press, London.
Bellwood, D.R., van Herwerden, L. & Konow, N. 2004. Evolu-
tion and biogeography of marine angelfishes (Pisces: Pom-
acanthidae). Mol. Phylogenet. Evol. 33: 140–155.
Bernardi, G., Bucciarelli, G., Costagliola, D., Robertson, D.R. &
Heiser, J.B. 2004. Evolution of coral reef fish Thalassoma spp.
(Labridae): 1. Molecular phylogeny and biogeography. Mar.
Biol. 144: 369–375.
Berumen, M.L. & Pratchett, M.S. 2006. Persistent disturbance
and long-term shifts in the structure of fish and coral
communities at Tiahura Reef, Moorea. Coral Reefs 25: 647–653.
Berumen, M.L. & Pratchett, M.S. 2008. Trade-offs associated with
dietary specialization in corallivorous butterflyfishes (Chae-
todontidae: Chaetodon). Behav. Ecol. Sociobiol. 62: 989–994.
Berumen, M.L., Pratchett, M.S. & McCormick, M.I. 2005.
Within reef variation in the diet and condition of two coral
feeding butterflyfish (Pices: Chaetodontidae). Mar. Ecol. Prog.
Ser. 287: 217–227.
Birkeland, C. & Neudecker, S. 1981. Foraging behavior of two
Caribbean chaetodontids: Chaetodon capistratus and C. aculeatus.
Copeia 1: 169–178.
Blot, J. 1969. Les poissons fossiles du Monte Bolca. Classe¢s
jusqu’ici dans les familles des Carangidae, Menidae, Ephippi-
dae, Scatophagidae. Mem. Mus. Civ. Stor. Nat. Verona 1: 1–525.
Blot, J. 1980. La faune ichthyologique des gisements du Monte
Bolca (Province de Verone, Italie): catalogue syste
´matique
pre
´sentant l’e
´tat actuel des recherches concernant cette faune.
Bull. Mus. Nat. Hist. Nat. 2: 339–396.
Blum, S.D. 1988. Biogeography of the Chaetodontidae: an
analysis of allopatry among closely related species. Environ.
Biol. Fish 25: 9–31.
Brown, J.W., Rest, J.S., Garcı´a-Moreno, J., Sorenson, M.D. &
Mindell, D.P. 2008. Strong mitochondrial DNA support for a
Cretaceous origin of modern avian lineages. BMC Biol. 6:6.
12 D. R. BELLWOOD ET AL.
ª2009 THE AUTHORS. J. EVOL. BIOL. doi:10.1111/j.1420-9101.2009.01904.x
JOURNAL COMPILATION ª2009 EUROPEAN SOCIETY FOR EVOLUTIONARY BIOLOGY

Carnevale, G. 2006. Morphology and biology of the Miocene
butterflyfish Chaetodon ficheuri (Teleostei: Chaetodontidae).
Zool. J. Linn. Soc. 146: 251–267.
Chow, S. & Hazama, K. 1998. Universal PCR primers for S7
ribosomal protein gene introns in fish. Mol. Ecol. 7: 1255–1256.
Coates, A.G., Jackson, J.B.C., Collins, L.S., Cronin, T.M., Dow-
sett, H.J., Bybell, L.M., Jung, P.J. & Obando, J.A. 1992. Closure
of the Isthmus of Panama: the nearshore record of Costa Rica
and western Panama. Geol. Soc. Am. Bull. 104: 814–828.
Cole, A.J., Pratchett, M.S. & Jones, G.P. 2008. Diversity and
functional importance of coral-feeding fishes on tropical coral
reefs. Fish Fish. 9: 286–307.
Cowman, P.F., Bellwood, D.R. & van Herwerden, L. 2009. Dating
the evolutionary origins of wrasse lineages and the rise of
trophic novelty on coral reefs. Mol. Phylogenet. Evol. 52: 621–631.
Drummond, A.J. & Rambaut, A. 2007. Bayesian evolution
analysis by sampling trees. BMC Evol. Biol. 7: 214.
Drummond, A.J., Ho, S.Y.W., Phillips, M.J. & Rambaut, A. 2006.
Relaxed phylogenetics and dating with confidence. PLoS Biol.
4: 699–710.
Dwyer, J., Li, H., Xu, D. & Liu, J.P. 2007. Transcriptional
regulation of telomerase activity: roles of the Ets transcription
factor family. Ann. N. Y. Acad. Sci. 1114: 36–47.
Elliott, J. & Bellwood, D.R. 2003. Alimentary tract morphology
and diet in three coral reef fish families. J. Fish Biol. 63: 1598–
1609.
Fauvelot, C., Bernardi, G. & Planes, S. 2003. Reductions in the
mitochondrial DNA diversity of coral reef fish provide
evidence of population bottlenecks resulting from Holocene
sea-level change. Evolution 57: 1571–1583.
Ferry-Graham, L.A., Wainwright, P.C. & Bellwood, D.R. 2001a.
Prey capture in long-jawedn butterflyfishes (Chaetodontidae):
the functional basis of novel feeding habits. J. Exp. Mar. Biol.
Ecol. 256: 167–184.
Ferry-Graham, L.A., Wainwright, P.C., Hulsey, C.D. & Bellwood,
D.R. 2001b. Evolution and mechanics of long jaws in
butterflyfishes (Family Chaetodontidae). J. Morphol. 248:
120–143.
Fessler, J.L. & Westneat, M.W. 2007. Molecular phylogenetics of
the butterflyfishes (Chaetodontidae): taxonomy and biogeog-
raphy of a global coral reef fish family. Mol. Phylogenet. Evol. 45:
50–68.
Findley, J.S. & Findley, M.T. 2001. Global, regional, and local
patterns in species richness and abundance of butterflyfishes.
Ecol. Monogr. 71: 69–91.
Floeter, S.R., Rocha, L.A., Robertson, D.R., Joyeux, J.C., Smith-
Vaniz, W.F., Wirtz, P., Edwards, A.J., Barreiros, J.P., Ferreira,
C.E.L., Gasparini, J.L., Brito, A., Falco
´n, J.M., Bowen, B.W. &
Bernardi, G. 2008. Atlantic reef fish biogeography and
evolution. J. Biogeogr. 35: 22–47.
Gregson, M., Berumen, M.L. & Pratchett, M.S. 2008. Bite rates
of butterflyfishes (Chaetodontidae) correlate with propor-
tional consumption of corals. Coral Reefs 27: 583–591.
Harmelin-Vivien, M.L. 2002. Energetics and fish diversity on
coral reefs. In: Coral Reef Fishes: Dynamics and Diversity in a
Complex Ecosystem (P.F. Sale, ed.), pp. 265–274. Academic
Press, New York.
Harmon, L.J., Weir, J.T., Brock, C.D., Glor, R.E. & Challenger,
W. 2008. GEIGER: investigating evolutionary radiations.
Bioinformatics 24: 129–131.
Ho, S.Y.W. 2007. Calibrating molecular estimates of substitution
rates and divergence times in birds. J. Avian Biol. 38: 404–414.
Hsu, K., Chen, J. & Shao, K. 2007. Molecular phylogeny of
Chaetodon (Teleostei: Chaetodontidae) in the Indo-West
Pacific: evolution in geminate species pairs and specific
groups. Raffles Bull. Zool. 14: 77–86.
Huelsenbeck, J.P. & Ronquist, F. 2001. MR. BAYES: Bayesian
inference of phylogenetic trees. Bioinformatics 17: 754–755.
Ihaka, R. & Gentleman, R. 1996. R: a language for data analysis
and graphics. J. Comput. Graph. Stat. 5: 299–314.
Irwin, D.M., Kocher, T.D. & Wilson, A.C.. 1991. Evolution of the
cytochrome b gene of mammals. J. Mol. Evol. 32: 128–144.
Johnson, K.G., Jackson, J.B.C. & Budd, A.F. 2008. Caribbean
reef development was independent of coral diversity over 28
million years. Science 319: 1521–1523.
Klanten, S.O., van Herwerden, L., Choat, J.H. & Blair, D. 2004.
Patterns of lineage diversification in the genus Naso (Acan-
thuridae). Mol. Phylogenet. Evol. 32: 221–235.
Kocher, T.D., Thomas, W.K., Meyer, A., Edwards, S.V., Paabo, S.,
Villablanca, F.X. & Wilson, C. 1989. Dynamics of mitochondrial
DNA evolution in animals: amplification and sequencing with
conserved primers. Proc. Natl Acad. Sci. 86: 6196–6200.
Konow, N. & Ferry-Graham, L. (in press). Functional morphol-
ogy of the butterflyfishes, In: The Biology of Butterflyfishes,
Chapter 2 (M.S. Pratchett, M.L. Berumen & B.G. Kapoor, eds).
Science Publishers Inc, Enfield, USA.
Konow, N., Bellwood, D.R., Wainwright, P.C. & Kerr, A.M.
2008. Evolution of novel jaw joints promote trophic diversity
in coral reef fishes. Biol. J. Linn. Soc. 93: 545–555.
Kuiter, R.H. 2002. Butterflyfishes, Bannerfishes and Their Relatives: A
Comprehensive Guide to Chaetodontidae and Microcanthidae. TMC
Publishing, Chorleywood.
Kulbicki, M., Bozec, Y.M. & Green, A. 2005. Implications of
biogeography in the use of butterflyfishes (Chaetodontidae) as
indicators for Western and Central Pacific areas. Aquat.
Conserv. Mar. Freshw. Ecosys. 15: S109–S126.
Lessios, H.A. 2008. The great American schism: divergence of
marine organisms after the rise of the Central American
Isthmus. Annu. Rev. Ecol. Evol. Syst. 39: 63–91.
Lessios, H.A., Kessing, B.D., Robertson, D.R. & Paulay, G. 1999.
Phylogeography of the pantropical sea urchin Eucidaris in
relation to land barriers and ocean currents. Evolution 53: 806–
817.
Littlewood, D.T.J., McDonald, S.M., Gill, A.C. & Cribb, T.H.
2004. Molecular phylogenetics of Chaetodon and the Chae-
todontidae (Teleostei: Perciformes) with reference to mor-
phology. Zootaxa 779: 1–20.
Lyons, L.A., Laughlin, T.F., Copeland, N.G., Jenkins, N.A.,
Womack, J.E. & O’Brien, S.J. 1997. Comparative anchor
tagged sequences (CATS) for integrative mapping of mamma-
lian genomes. Nat. Genet. 15: 47–56.
Maddison, W. & Maddison, D. 2007. Mesquite v 2.01. URL
http://mesquiteproject.org.
Magallon, S. & Sanderson, M.J. 2001. Absolute diversification
rates in angiosperm clades. Evolution 55: 1762–1780.
Maguire, B.A. & Zimmermann, R.A. 2001. The ribosome in
focus. Cell 104: 813–816.
McCafferty, S., Bermingham, E., Quenouille, B., Planes, S.,
Hoelzer, G. & Asoh, K. 2002. Historical biogeography and
molecular systematics of the Indo-Pacific genus Dascyllus
(Teleostei: Pomacentridae). Mol. Ecol. 11: 1377–1392.
McIlwain, J.L. & Jones, G.P. 1997. Prey selection by an obligate
coral-feeding wrasse and its response to small-scale distur-
bance. Mar. Ecol. Prog. Ser. 155: 189–198.
Origins of coral feeding 13
ª2009 THE AUTHORS. J. EVOL. BIOL. doi:10.1111/j.1420-9101.2009.01904.x
JOURNAL COMPILATION ª2009 EUROPEAN SOCIETY FOR EVOLUTIONARY BIOLOGY

McMillan, W.O. & Palumbi, S.R. 1995. Concordant evolutionary
patterns among Indo-West Pacific butterflyfishes. Proc. R. Soc.
Lond. B 260: 229–236.
Motta, P.J. 1988. Functional morphology of the feeding appa-
ratus of ten species of Pacific butterflyfishes (Perciformes,
Chaetodontidae): an ecomorphological approach. Environ.
Biol. Fish 22: 39–67.
Nee, S., Mooers, O. & Harvey, P.H. 1992. Tempo and mode of
evolution revealed from molecular phylogenies. Proc. Natl
Acad. Sci. USA 89: 8322–8326.
Nylander, J.A.A. 2004. MrModeltest v.2. Evolutionary Biology
Centre, Uppsala University, Uppsala.
Nylander, J.A.A., Ronquist, F., Huelsenbeck, J.P. & Nieves-
Aldrey, J.L. 2004. Bayesian phylogenetic analysis of combined
data. Syst. Biol. 53: 47–67.
Palumbi, S.R., Grabowsky, G., Duda, T., Geyer, L. & Tachino, N.
1997. Speciation and population genetic structure in tropical
Pacific sea urchins. Evolution 51: 1506–1517.
Patterson, C. 1993. Osteichthyes: Teleostei. In: The Fossil Record 2
(M.J. Benton, ed.), pp. 621–656. Chapman and Hall, London.
Posada, D. & Crandall, K.A. 1998. Modeltest: testing the model
of DNA substitution. Bioinformatics 14: 814–818.
Pratchett, M.S. 2005. Dietary overlap among coral-feeding
butterflyfishes (Chaetodontidae) at Lizard Island, northern
Great Barrier Reef. Mar. Biol. 148: 373–382.
Pratchett, M.S. 2007. Dietary selection by coral-feeding butter-
flyfishes (Chaetodontidae) on the Great Barrier Reef, Austra-
lia. Raffles Bull. Zool. 14: 155–160.
Pratchett, M.S., Wilson, S.K., Berumen, M.L. & McCormick, M.I.
2004. Sub-lethal effects of coral bleaching on an obligate coral
feeding butterflyfish. Coral Reefs 23: 352–356.
Pratchett, M.S., Wilson, S.K. & Baird, A.H. 2006. Declines in
the abundance of Chaetodon butterflyfishes (Chaetodontidae)
following extensive coral depletion. J. Fish Biol. 69: 1269–
1280.
Pratchett, M.S., Munday, P.L., Wilson, S.K., Graham, N.A.J.,
Cinner, J.E., Bellwood, D.R., Jones, G.P., Polunin, N.V.C. &
McClanahan, T.R. 2008. Effects of climate-induced coral
bleaching on coral-reef fishes: ecological and economic con-
sequences. Oceanogr. Mar. Biol. Ann. Rev. 46: 251–296.
Pybus, O.G. & Harvey, P.H. 2000. Testing macro-evolutionary
models using incomplete molecular phylogenies. Philos. Trans.
R. Soc. (Lond.) B 267: 2267–2272.
Rabosky, D.L. 2006. LASER: a maximum likelihood toolkit for
detecting temporal shifts in diversification rates from molec-
ular phylogenies. Evol. Bioinform. Online 2: 247–250.
Rambaut, A. 1996. Se-Al: sequence alignment editor. URL
http://evolve.zoo.ox.ac.uk.
Rambaut, A. & Drummond, A.J. 2007. Tracer v1.4. URL
http://beast.bio.ed.ac.uk/Tracer.
Read, C.I., Bellwood, D.R. & van Herwerden, L. 2006. Ancient
origins of Indo-Pacific coral reef fish biodiversity: a case study
of the leopard wrasses (Labridae: Macropharyngodon). Mol.
Phylogenet. Evol. 38: 808–819.
Reese, E.S. 1975. A comparative field study of the social
behavior and related ecology of reef fishes of the family
Chaetodontidae. Z. Tierpsychol. 37: 37–61.
Renema, W., Bellwood, D.R., Braga, J.C., Bromfield, K., Hall, R.,
Johnson, K.G., Lunt, P., Meyer, C.P., McMonagle, L., Morley,
R.J., O’dea, A., Todd, J.A., Wesselingh, F.P., Wilson, M.E.J. &
Pandolfi, J.M. 2008. Hopping hotspots: global shifts in marine
biodiversity. Science 321: 654–657.
Roberts, C.M., Ormond, R.F.G. & Shepherd, A.R.D. 1988. The
usefulness of butterflyfishes as environmental indicators on
coral reefs. In: Proceedings of the 6th International Coral Reef
Symposium 2 (J.H. Choat, ed.), pp. 331–336. Townsville,
Australia.
Robertson, D.R., Krag, F., de Moura, R.L., Victor, B.C. & Bernardi,
G. 2006. Mechanisms of speciation and faunal enrichment in
Atlantic parrotfishes. Mol. Phylogenet. Evol. 40: 795–807.
Rotjan, R.D. & Lewis, S.M. 2008. Impact of coral predators on
tropical reefs. Mar. Ecol. Prog. Ser. 367: 73–91.
Sambrook, J. & Russell, D.W. 2001. Molecular Cloning, a Labora-
tory Manual. Cold Spring Harbour Laboratory Press, New York.
Sanderson, M.J. 2002. Estimating absolute rates of molecular
evolution and divergence times: a penalized likelihood
approach. Mol. Biol. Evol. 19: 101–109.
Sanderson, M.J. 2004. r8s, Version 1.70: User’s Manual. Section of
Evolution and Ecology, University of California, Davis, CA.
Smith, W.L. & Wheeler, W.C. 2006. Venom evolution wide-
spread in fishes: a phylogenetic road map for the bioprospect-
ing of piscine venoms. J. Hered. 97: 206–217.
Smith, W.L., Webb, J.F. & Blum, S.D. 2003. The evolution of the
laterophysic connection with a revised phylogeny and taxon-
omy of butterflyfishes (Teleostei: Chaetodontidae). Cladistics
19: 287–306.
Steininger, F.F. & Ro
¨gl, F. 1984. Paleography and palinspastic
reconstruction of the Neogene of the Mediterranean and
Paratethys. In: The Geological Evolution of the Eastern Mediterra-
nean (J.E. Dixon & A.H. Robertson, eds), pp. 659–668.
Blackwell, Oxford.
Streelman, J.T., Alfaro, M., Westneat, M.W., Bellwood, D.R. &
Karl, S.A. 2002. Evolutionary history of the parrotfishes:
biogeography, ecomorphology, and comparative diversity.
Evolution 56: 961–971.
Swofford, D.L. 1998. PAUP* Phylogenetic Analysis Using Parsimony
(* and Other Methods). Sinauer Associates, Sunderland, MA.
Thompson, J.D., Gibson, T.J., Plewniak, F.J. & Higgins, D.G.
1997. The ClustalX windows interface: flexible strategies for
multiple sequence alignment aided by quality analysis tools.
Nucleic Acids Res. 24: 4876–4882.
Tyler, J.C. & Sorbini, C. 1999. Phylogeny of the fossil and recent
genera of fishes of the family Scatophagidae (Squamipinnes).
Boll. Mus. Civ. Stor. Nat. Verona 23: 353–393.
Vermeij, G.J. 1977. The mesozoic marine revolution: evidence
from snails, predators and grazers. Paleobiology 3: 245–258.
Wallace, C.C. & Rosen, B. 2006. Diverse staghorn corals
(Acropora) in high-latitude Eocene assemblages: implications
for the evolution of modern diversity patterns of reef corals.
Proc. Biol. Sci. 273: 975–982.
Westneat, M.W. & Alfaro, M. 2005. Phylogenetic relationships
and evolutionary history of the reef fish family Labridae. Mol.
Phylogenet. Evol. 36: 370–390.
Wilson, M.E.J. 2008. Global and regional influences on equa-
torial shallow-marine carbonates during the Cenozoic. Palae-
ogeogr. Palaeoclimatol. Palaeoecol. 265: 262–274.
Wilson, S.K., Graham, N.A.J., Pratchett, M.S., Jones, G.P. &
Polunin, N.V.C. 2006. Multiple disturbances and the global
degradation of coral reefs: are reef fishes at risk or resilient?
Glob. Chang. Biol. 12: 2220–2234.
Wylie, C.R. & Paul, V.J. 1989. Chemical defenses in three species
of Sinularia (Coelenterata, Alcyonacea): effects against gener-
alist predators and the butterflyfish Chaetodon unimaculatus
Bloch. J. Exp. Mar. Biol. Ecol. 129: 141–160.
14 D. R. BELLWOOD ET AL.
ª2009 THE AUTHORS. J. EVOL. BIOL. doi:10.1111/j.1420-9101.2009.01904.x
JOURNAL COMPILATION ª2009 EUROPEAN SOCIETY FOR EVOLUTIONARY BIOLOGY

Xia, X.H. & Xie, Z. 2001. DAMBE: data analysis in molecular
biology and evolution. J. Hered. 92: 371–373.
Xia, X.H., Xie, Z., Salemi, M., Chen, L. & Wang, Y. 2003. An
index of substitution saturation and its application. Mol.
Phylogenet. Evol. 26: 1–7.
Yang, Z. & Rannala, B. 2006. Bayesian estimation of species
divergence times under a molecular clock using multiple
fossil calibrations with soft bounds. Mol. Biol. Evol. 23: 212–
226.
Zekeria, Z.A., Dawit, Y., Ghebremedhin, S., Naser, M. & Videler,
J.J. 2002. Resource partitioning among four butterflyfish
species in the Red Sea. Mar. Freshw. Res. 53: 63–168.
Zwickl, D.J. 2006. Garli – genetic algorithm for rapid likelihood
inference. URL http://www.bio.utexas.edu/faculty/antisense/
garli/Garli.html.
Supporting information
Additional Supporting Information may be found in the
online version of this article:
Appendix S1 Table1: List of species in the Chaetodon-
tidae; their taxonomy, diet and habitat associations.
Table 2: Primer sequences.
Appendix S2 Estimated ages of nodes with HPD
distributions.
Figure S1 Coral reef use as habitat preference and the
likelihood (ML) of the presence of this triat in the most
recent common ancestor (mrca) of each clade, as indi-
cated by the pie charts at each node, when mapped to the
best phylogenetic reconstruction of the group.
Figure S2 Coravllivory and the likelihood (ML) of the
presence of this trait in the most recent common ancestor
(mrca) of each clade, as indicated by the pie charts at
each node, when mapped to the best phylogenetic
reconstruction of the group.
As a service to our authors and readers, this journal
provides supporting information supplied by the authors.
Such materials are peer-reviewed and may be re-
organized for online delivery, but are not copy-edited
or typeset. Technical support issues arising from support-
ing information (other than missing files) should be
addressed to the authors.
Received 20 October 2009; accepted 23 October 2009
Origins of coral feeding 15
ª2009 THE AUTHORS. J. EVOL. BIOL. doi:10.1111/j.1420-9101.2009.01904.x
JOURNAL COMPILATION ª2009 EUROPEAN SOCIETY FOR EVOLUTIONARY BIOLOGY