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ORIGINAL
ARTICLE
Biogeography of the Mesoamerican
Cichlidae (Teleostei: Heroini):
colonization through the GAARlandia
land bridge and early diversification
Oldr
ˇich R
ˇı
´c
ˇan
1
*, Lubomı
´r Pia
´lek
1
, Rafael Zardoya
2
, Ignacio Doadrio
2
and Jan Zrzavy
´
1
1
Department of Zoology, University of South
Bohemia, C
ˇeske
´Bude
ˇjovice, Czech Republic,
2
Departamento de Biodiversidad y Biologı
´a
Evolutiva, Museo Nacional de Ciencias
Naturales, Madrid, Spain
*Correspondence: Oldr
ˇich R
ˇı
´c
ˇan, Department
of Zoology, Faculty of Science, University of
South Bohemia, Branis
ˇovska
´31, 37005, C
ˇeske
´
Bude
ˇjovice, Czech Republic.
E-mail: oldrichrican@yahoo.com
ABSTRACT
Aim We present a molecular phylogenetic and biogeographical analysis of the
Mesoamerican cichlid fishes (Cichlidae: Cichlasomatinae: Heroini), a dominant
part of the freshwater biodiversity of the region. Based on these analyses we
investigate the spatial and temporal origins and diversification of the group.
Location Mesoamerica.
Methods Model-based phylogenetic methods (MrBayes) using seven molecu-
lar markers with a virtually complete species-level taxon sampling, together
with the Bayesian approach to statistical dispersal–vicariance analysis (S-DIVA),
and fossil- and palaeogeography-calibrated beast molecular clock analyses were
used to assess the timing of dispersal, vicariance and diversification events.
Results We present a robust multigene phylogeny resolved from the deepest
nodes to the species level. Two colonizations of Middle America and one of
the Greater Antilles occurred from South America within a narrow time
window during the Oligocene epoch. Cichlid colonization of Central America
proceeded from the north in the Early–Middle Miocene. Central America later
became repeatedly fragmented during the latter half of the Miocene, which led
to the formation of the present ichthyological provinces prior to the final
closure of the Panama Isthmus.
Main conclusions The heroine cichlid fishes colonized the Greater Antilles
and Middle America simultaneously through the GAARlandia land bridge
during the Oligocene. Central America (including eastern Panama) was colo-
nized from northern Middle America in the Early–Middle Miocene. Thus our
results do not support a direct colonization of Middle America from South
America, or a Cretaceous–Palaeocene colonization through the proto-Antilles,
or a colonization of the Greater Antilles from Middle America as suggested by
previous studies.
Keywords
Central America, freshwater fishes, Greater Antilles, ichthyological provinces,
Middle America, molecular clocks, molecular phylogeny, plate tectonics,
statistical dispersal–vicariance analysis.
INTRODUCTION
Mesoamerica is a highly complex part of the Earth’s crust,
composed of a dynamic pattern of tectonic plates and
terranes. Its relatively small size but extremely complicated
geological history, together with its relative isolation
throughout most of the Cenozoic, is reflected in its high bio-
logical diversity and peculiarities of its faunal composition.
The majority of taxa in Mesoamerica are of Neotropical
origin, but composition of the Mesoamerican fauna is signifi-
cantly different from that of South America (the core of the
Neotropical realm), the former being conspicuous for its
limited higher-order taxonomic composition. Among the
freshwater fish groups, those dominant in South America
(primary freshwater fishes; mainly Ostariophysi) are only
marginal in Mesoamerica, while other, secondary freshwater
ª2012 Blackwell Publishing Ltd http://wileyonlinelibrary.com/journal/jbi 579
doi:10.1111/jbi.12023
Journal of Biogeography (J. Biogeogr.) (2013) 40, 579–593
fishes are less significant in South America but completely
dominate Mesoamerica (Poeciliidae, Cichlidae; Myers, 1949,
1966; Bussing, 1976, 1985).
Myers (1966) and Bussing (1976, 1985) were the first to
propose an explanation for this freshwater fish composition
of Mesoamerica. Bussing (1985) postulated that an ancient
South American element (secondary freshwater fishes) dis-
persed into Middle America (i.e. tropical Mexico to eastern
Panama) during the Late Cretaceous or the Palaeocene, and
that Middle America was later isolated from South America
during most of the Tertiary by a marine seaway through
lower Central America until the final emergence of the
Panama Isthmus, which enabled colonization by primary
freshwater fishes. Bussing, in agreement with Myers (1966)
and Miller (1966), explained the composition of the Middle
American ichthyofauna as being derived from different dis-
persal capabilities of primary and secondary freshwater fishes.
He thus also proposed a considerably ancient origin for part
of the Middle American fish fauna.
Based on information of Caribbean plate tectonics
available in the 1970s, Rosen (1975) developed a vicariance
model of Caribbean biogeography. This model postulated a
wide gap in space and time between the existence of the
Cretaceous–Palaeocene palaeo-Antillean land bridge and the
late Cenozoic Middle American land bridge. More recent
studies have, however, demonstrated that the Greater Antilles
were probably connected with South America at least once
in the mid-Cenozoic [around the Eocene–Oligocene transi-
tion, 35–33 million years ago (Ma)] through the GAARlandia
land bridge (Iturralde-Vinent & MacPhee, 1999). Studies on
fossil mammals of strictly North American affinities collected
in central Panama (Kirby & MacFadden, 2005; MacFadden,
2006; Kirby et al., 2008) also suggest that around 19.5–
14 Ma (Middle Miocene) Middle America was a continuous
peninsula of North America (or a succession of insular step-
ping stones; Coates & Obando, 1996; Coates et al., 2004).
These studies thus propose a land bridge (or successively
connected stepping stones) from South America to the
Greater Antilles through the now submerged Aves Ridge
(Lesser Antilles) at the Eocene–Oligocene transition. Later, in
the Miocene, a land bridge (or stepping stones) existed
between North America and Panama which was not
connected to South America until the Great American Inter-
change (GAI). To hypothesize an overland colonization route
of Middle America from South America during the mid-
Cenozoic, the narrow sea strait between the developing
island of Cuba and the Maya (Yucatan) and Chortis
(Honduras–Nicaragua) geological terranes is the only marine
barrier remaining that would have to be crossed. Could it
indeed be that some degree of salinity tolerance enabled
colonization of Middle America by secondary freshwater
fishes prior to the GAI (Myers, 1949, 1966; Miller, 1966;
Bussing, 1976, 1985)?
The three most plausible scenarios for the colonization of
Mesoamerica in general and Middle America in particular
are thus as follows.
1. The old Cretaceous–Palaeocene proto-Antilles land
bridge.
2. The mid-Cenozoic Greater Antilles–Aves Ridge land
bridge (GAARlandia) plus the Miocene Middle American
land bridge with a narrow gap between the Greater Antilles
and Middle America.
3. Direct, probably multiple and independent overwater
colonization of the Greater Antilles and of Middle America.
In the case of the Cichlidae, various authors have either
supported a mid-Cenozoic (Bermingham & Martin, 1998;
Concheiro Pe
´rez et al., 2007; Hulsey et al., 2010, 2011) or a
Cretaceous–Palaeocene colonization (Chakrabarty, 2006;
Chakrabarty & Albert, 2011). However, although the Meso-
american heroine cichlids have been the subject of compre-
hensive phylogenetic research (Bermingham & Martin, 1998;
Chakrabarty, 2006; Concheiro Pe
´rez et al., 2007; R
ˇı
´c
ˇan et al.,
2008; Hulsey et al., 2010, 2011; Chakrabarty & Albert, 2011),
a convincing biogeographical synthesis for the Mesoamerican
heroine cichlids, based on complete taxon sampling free of
misidentification, multigene analyses and molecular clock
and historical-biogeography analyses, is still pending.
The biogeographical diversification of cichlids within
Middle America has received far less attention than the colo-
nization of the area. This is quite surprising, given that more
than 120 Middle American species are endemics, and cichlid
diversity and genus-level taxonomy in Middle America is
one of the most complex in the whole of the Neotropics
(Concheiro Pe
´rez et al., 2007; R
ˇı
´c
ˇan et al., 2008). However,
the origins of this diversity remain virtually unstudied except
for local studies (Barluenga & Meyer, 2004; Hulsey et al.,
2004; Barluenga et al., 2006; Geiger et al., 2010).
Understanding how the community of terrestrial and fresh-
water vertebrates in Mesoamerica became established is one of
the most intriguing challenges in biogeography. Our study
seeks to add to the discussion by focusing on the following
two goals: (1) to reconstruct the colonization of Mesoamerica
by heroine cichlid fishes; and (2) to complement the coloniza-
tion scenario with a reconstruction of their biogeographical
evolution within Middle America. For these goals we use phy-
logenetic analyses that include several molecular markers and
a virtually complete taxon sampling throughout Mesoamerica
and northern South America, together with a molecular clock
analysis calibrated by well-established geological events and
fossils, and complete distribution records of 116 species
scored for all 12 recognized ichthyological provinces (IPs).
MATERIALS AND METHODS
Phylogenetic tree of the Mesoamerican Cichlidae
Neotropical Cichlidae are divided into several clades, among
which the largest ones are classified as the subfamilies Geo-
phaginae and Cichlasomatinae. All but two cichlid species in
Mesoamerica belong to the tribe Heroini of the subfamily
Cichlasomatinae (e.g. Farias et al., 1999, 2000, 2001; Sparks
& Smith, 2004; Smith et al., 2008; Lo
´pez-Ferna
´ndez et al.,
Journal of Biogeography 40, 579–593
ª2012 Blackwell Publishing Ltd
580
O. R
ˇı
´c
ˇan et al.
2010). The Heroini further include several South American
genera (Australoheros,Caquetaia,Heroina,Heros,Hoplarchus,
Hypselecara,Mesonauta,Pterophyllum,Symphysodon,Uaru,
and the unnamed ‘Heros’ festae group genus).
The data set used in the present study was combined from
several recently published phylogenetic studies focusing on
Mesoamerican heroine cichlids and several new sequences.
The data set includes all lineages (genera) and all valid
species of the Mesoamerican heroines, with only a few excep-
tions, and we have excluded several previously used sequences
that are actually cases of species misidentification (see Appen-
dix S1 in Supporting Information). In total, the data set
includes 123 heroine species. Two cichlasomatines (represent-
ing the sister group of the Heroini: Cichlasoma,Andinoacara)
and one geophagine (Satanoperca) were used as outgroups.
The data set includes seven molecular markers, four
mitochondrial [cytochrome b(cyt b), NADH dehydrogenase
subunit 4 (ND4), cytochrome coxidase subunit I (COI), 16S
rRNA (16S)] and three nuclear [RAG1,RAG2, S7 intron 1
(S7i1)], which all have a representative coverage across the
genus-level diversity of the studied group. The mitochondrial
DNA (mtDNA) cyt bgene is sampled for all terminal taxa
(species).
DNA isolation, polymerase chain reaction (PCR) and
sequencing
The markers newly sequenced in this study included the mito-
chondrial cyt band the nuclear RAG1 and S7i1. Genomic
DNA was extracted from ethanol-preserved gill or fin tissue
using the JETQUICK Tissue DNA Spin Kit (Genomed, Lo
¨hne,
Germany) following standard protocol. The primers and reac-
tion conditions of PCR amplification for all loci are as in
R
ˇı
´c
ˇan et al. (2008). Each PCR reaction volume of 25 lL con-
tained 12.5 lL of Combi PPP Master Mix (Top-Bio; http://
www.top-bio.cz/), 1.5 lL of each primer (10 pmol lL
1
),
and 1 lL of extracted DNA. The PCR reactions were per-
formed in a Bioer XP Thermal Cycler (Hangzhou, P.R. China)
and PCR products were purified using the JETQUICK PCR
Purification Spin Kit (Genomed). Sequencing reactions were
performed following standard protocol with the use of the
same primers as in R
ˇı
´c
ˇan et al. (2008), and the products were
analysed in an ABI 3730XL automated sequencer (Applied
Biosystems, Foster City, CA, USA; both steps done by Macro-
gen Inc., Seoul, Korea). Contiguous sequences of the gene seg-
ments were created by assembling DNA strands (forward and
reverse) using BioLign 4.0.6.2 (Hall, 2001). Nucleotide coding
sequences were also translated into protein sequences to check
for possible stop codons or other ambiguities. All sequences
were submitted to GenBank under accession numbers
HQ197729 and JX437626–JX437640.
Alignment
Sequences were aligned with muscle 3.8 (Edgar, 2004),
using the default settings. The 16S and S7i1 markers were
additionally realigned (option ‘refine’; no subjective ‘by-eye’
treatment was applied). Gaps (indels) were kept as poten-
tially informative characters for these two loci. Separate
alignments of individual loci were assembled into a final
combined matrix by a computer program created in Borland
Delphi (Borland Delphi for Microsoft Windows, version 10,
2005, Borland Software Corporation, Austin, TX, USA),
written by the second author.
Phylogenetic methods
MrBayes 3.1.2 (Huelsenbeck & Ronquist, 2001; Ronquist &
Huelsenbeck, 2003) was used for the Bayesian inference of
phylogeny. The seven molecular markers used were analysed
using three different data partition schemes: all markers
individually (seven partitions), mitochondrial and nuclear
separately, both divided into coding and non-coding (four
partitions) and same as the latter with mitochondrial coding
loci divided by codon position (first plus second versus
third; five partitions). An optimal model of evolution for
each data partition according to Akaike’s information crite-
rion was selected using MrModelTest 2.2 (Nylander, 2004)
and paup* 4.0b10 (Swofford, 2003). The Bayesian analysis
using the Markov chain Monte Carlo (MCMC) simulation
was run with unlinked parameters (except for branch length
and topology) for 10 million generations with trees sampled
and saved every 1000 generations. Two independent analyses,
each comprising two runs with eight chains, were performed
to compare results of independent analyses. The analyses
were run at the freely available Bioportal server (http://www.
bioportal.uio.no/). The first 25% of trees from each run were
discarded as burn-in. Convergence of the runs was estimated
with the use of: (1) diagnostic criteria produced by the
‘sump’ command in MrBayes; (2) graphical visualization
and diagnostics in Tracer 1.5.0 (Rambaut & Drummond,
2007). The remaining trees were used for reconstruction of
the 50% majority-rule consensus tree with posterior proba-
bility (PP) values of the branches.
Biogeographical methods
The study covers all areas of Mesoamerica, a combined area
including Middle America (i.e. geological North America from
tropical Mexico to Guatemala, plus Central America, the latter
further subdivided into nuclear Central America from
Honduras to Nicaragua, and lower Central America from
Costa Rica to eastern Panama), the Greater Antilles (Cuba,
Hispaniola) and trans-Andean north-western South America
(Pacific coast of Colombia, Ecuador, and northernmost Peru).
Middle America is, based on species distribution patterns,
divided into 12 IPs, six on the Caribbean versant and six on
the Pacific one (Miller, 1966; Bussing, 1976, 1985; Miller
et al., 2005; Smith & Bermingham, 2005). The highest diver-
sity is found in two IPs on the broad Caribbean versant: (1)
the Usumacinta province (southern Mexico to Guatemala)
with at least 46 cichlid species; and (2) the San Juan
Journal of Biogeography 40, 579–593
ª2012 Blackwell Publishing Ltd
581
Biogeography of Mesoamerican Cichlidae
province (northern Nicaragua to eastern Costa Rica) with at
least 28 cichlid species. All IPs of Middle America which are
naturally inhabited by the heroine cichlids were included. In
total, 14 distribution areas (corresponding to the IPs in Mid-
dle America plus adjacent areas) were used for coding the
distribution records of all species for the biogeographical
(statistical dispersal–vicariance, S-DIVA) analysis. Sources of
distribution records for all included heroine cichlid species
in Mesoamerica are listed in Appendix S1.
In order to elucidate ancestral areas for all nodes of the
phylogenetic tree, we used the event-based Bayesian S-DIVA
analysis (implemented in rasp 2.0; Yu et al., 2011). Distribu-
tions of all species at the level of IPs were input into
S-DIVA. Rather than using a single fully resolved topology,
which often is possible only by making a priori assumptions
about relationships within a given tree set, S-DIVA takes
topological uncertainty into account. For the S-DIVA analy-
ses we used Bayesian sampled trees from MrBayes runs with
10 million generations with every 1000th tree sampled and
25% trees discarded as burn-in. The probability of ancestral
areas for nodes was then plotted on the majority-rule con-
sensus tree derived from MCMC. The analysis was carried
out using the ‘maxareas =4’ option in S-DIVA. This is
equivalent to assuming that the ancestors of the group in
question have the same ability to disperse as their extant
descendants and that ancestral ranges were therefore similar
in size to extant ranges (Sanmartı
´n, 2003; Nylander et al.,
2008).
Molecular clock and fossil calibration of
Mesoamerican cichlids
We used the Bayesian Evolutionary Analysis by Sampling
Trees (beast) software package version 1.6.1 (Drummond &
Rambaut, 2007) to estimate the divergence dates within the
Mesoamerican Cichlidae under the relaxed molecular clock
model with uncorrelated lognormal distribution of rates. We
set a Yule speciation process for the tree prior (Drummond
et al., 2006). We assigned the best-fitting model, as estimated
by MrModeltest 2.2 (Nylander, 2004), to each of the data
partitions (using the three data partition schemes described
in MrBayes runs above). The analyses in beast were per-
formed with several independent runs for 30 million genera-
tions with trees sampled every 10,000 generations. The
analyses were run at the freely available Bioportal server
(www.bioportal.uio.no). Runs were checked for convergence
diagnostics with Tracer 1.5.0. Four well-converged runs
were combined in LogCombiner 1.6.1 with a burn-in of
25% for each of the data partition schemes. The final tree
for each data partition scheme was produced from these data
with TreeAnotator 1.6.1.
We used default prior distributions for all parameters
except for calibration points derived from fossils and well-
established geological vicariant events. The calibration points
defined in BEAUti 1.6.1 (Drummond & Rambaut, 2007)
were as follows.
1. The minimum age of the fossil Plesioheros chauliodus (at
39.9–48.6 Ma; BEAUti input, mean 44.25 Ma with
SD = 2.7) as the time for the divergence of the former from
its sister group (all remaining heroine cichlids, except Hyp-
selecara plus Hoplarchus; Perez et al., 2010).
2. The split between Cuba and Hispaniola at 20–25 Ma
(end of the Oligocene; BEAUti input, mean 22.5 Ma with
SD = 1.5) as the mean for the divergence between Cuban
(Nandopsis tetracanthus plus Nandopsis ramsdeni) and Hispa-
niolan (Nandopsis haitiensis) species (R
ˇı
´c
ˇan et al., 2008; cf.
21 Ma, Pindell, 1994; 24–23 Ma, Iturralde-Vinent & Mac-
Phee, 1999; 24 Ma, Da
´valos, 2004; 25–27 Ma, Roca et al.,
2004; 20–25 Ma, Chakrabarty, 2006), further supported by
the minimum age of the Hispaniolan fossil Nandopsis woodr-
ingi (minimum age 15 Ma; Tee-Van, 1935; Chakrabarty,
2006).
3. The separation of the Orinoco and Magdalena drainage
basins by the final rise of the Cordillera Oriental (10.1–
11.8 Ma; Lundberg et al., 1998; BEAUti input, mean
10.95 Ma with SD = 0.6) as the age of divergence between
Caquetaia sp. cf. kraussii and Caquetaia spectabile.
Palaeogeographical maps
Palaeogeographical maps of subaerial land configuration were
reconstructed on the basis of the geological literature
(Appendix S1).
RESULTS
Alignment of coding sequences exhibited no stop codons or
other inconsistencies. The informativeness of individual
markers and the chosen models under different data parti-
tioning are reported in Table 1. Phylogenetic analyses per-
formed on the concatenated data set (cyt b,ND4,COI,16S,
RAG1,RAG2,S7i1) using Bayesian inference provided robust
and highly congruent results using the three different data
partition schemes (see Table 1). Runs in independent analy-
ses for each partition scheme converged well, and indepen-
dent analyses of each partition scheme led to virtually
identical topologies. The differences in topology and poster-
ior probabilities between the three partition schemes were
negligible. The highest sum of significant posterior probabili-
ties (PP =0.95 or higher) was attained by the five-partition
analysis which we took as our final phylogenetic tree (Fig. 1).
Effective sample size (ESS) values as reported in Tracer
1.5.0 for both runs combined and using a 25% burn-in were
higher than 200 for all parameters.
All internal nodes relevant to the reconstruction of histori-
cal biogeography in Mesoamerican heroine cichlids have a
posterior probability of 1 (Figs 1 & 2). The Mesoamerican
heroine cichlids form a well-supported clade in this tree
(Fig. 1). They are further divided into two major clades. The
herichthyine clade is completely Middle American, except for
one secondarily South American lineage (the ‘Heros’ festae
group). The other clade contains the basal and southernmost
Journal of Biogeography 40, 579–593
ª2012 Blackwell Publishing Ltd
582
O. R
ˇı
´c
ˇan et al.
South American heroine cichlids (Australoheros), a subclade
grouping another South American lineage (Caquetaia,Hero-
ina) with the Greater Antillean Nandopsis, and as a third
subclade the strictly Middle American amphilophine cichlids.
Most nodes of the herichthyine clade are strongly supported.
On the contrary, the basal nodes of the amphilophines lack
significant support, which is accompanied by very short
internodal branches. The amphilophine cichlid clade also
shows a remarkably long basal branch, suggesting a long-
term lack of diversification, or a significant number of
extinctions.
The time-calibrated phylogeny of the concatenated data
set (cyt b,ND4,COI,16S,RAG1,RAG2,S7i1) using beast
provided robust and highly congruent results using the
three different data partition schemes (see Table 1). The five-
partition analysis had the highest ESS scores (using a 25%
burn-in) and is shown in Fig. 2. All ESS scores in this
analysis were above 200, with the ESS score for posterior
probability equal to 546. The difference in age estimates at
the key nodes for the colonization of Mesoamerica (which
span 35.9–24.2 Ma in the final five-partition analysis; Fig. 2)
was only 0.4–0.2 Myr between the three different analyses.
Also, analyses run using each of the calibration points by
itself (see Materials and Methods) provided very similar age
estimates for the colonization of Mesoamerica, differing only
by 1–2 Myr, which suggests lack of conflict between the
calibration points. Trimming of outgroup taxa did not sig-
nificantly change age estimates.
Based on our results (Fig. 2), the colonization of Mesoamer-
ica by heroine cichlid fishes occurred within a relatively nar-
row time window during the Oligocene and is congruent with
the period of existence of the GAARlandia land bridge. The
colonization is reconstructed as composed of two instances of
colonization of Middle America and one of the Greater Antil-
les, all from South America, plus one back-colonization of
South America from Middle America (Figs 2 & 3a).
1. The oldest colonization occurred between 35.9 and
31.8 Ma [mean values: 40.1–27.9 Ma, 95% highest posterior
density (HPD); Fig. 2] and to the present this lineage
survived as the first main clade of Middle American cichlids,
the herichthyine cichlids (Fig. 1), ancestrally endemic to the
Maya terrane (Fig. 2).
2. The second colonization included the second main clade
of Middle American cichlids, the amphilophine cichlids
(Fig. 1), ancestrally endemic to the Chortis terrane (Fig. 2),
together with the Antillean Cichlidae (Nandopsis), and
occurred between 29.6 and 28.0 Ma (mean values: 33.0–
24.6 Ma, 95% HPD). This colonization occurred in a clade
that also includes South American taxa (Caquetaia,Heroina,
Australoheros) which suggests that the first colonization
by the herichthyine cichlids was indeed a separate, slightly
earlier event.
3. The last dispersal event through the GAARlandia land
bridge was the back-colonization of South America from
Middle America between 28.9 and 26.3 Ma (mean values:
32.6–22.9 Ma, 95% HPD), followed by the vicariance of the
South American ‘Heros’ festae group from its Middle Ameri-
can sister group (at 24.2 Ma, mean: 27.8–20.9 Ma, 95%
HPD). This event marks the termination of the reconstructed
duration of the GAARlandia land bridge (Figs 2 & 3a,b).
The two Middle American cichlid clades (the herichthyines
and the amphilophines; Fig. 1) are reconstructed as having
had exclusive ancestral areas in Middle America (Figs 2, 3a,b
& 4a), and this past distribution pattern is still evident in the
present fauna. The herichthyine cichlids originated in and
dominate the Usumacinta province (B) of the Maya terrane
of North America, while the amphilophines originated in the
San Juan province (C) of the Chortis terrane of Middle
America and dominate Central America (Figs 2, 3a,b & 4a).
The first Middle American clade (the herichthyines) is
composed of four main subclades. The crown-group (Herich-
thys,Paraneetroplus,Theraps,‘Heros’ grammodes,Thorichthys,
‘Heros’ salvini) is endemic to the Usumacinta province, with
a basal divergence at 22.9 Ma (26.3–19.6 Ma, 95% HPD;
Fig. 2). Its sister group (the unnamed ‘Heros’ festae-group
genus) has a disjunct distribution in north-western South
America due to back-colonization of South America (see
above). The third subclade of the herichthyines, Herotilapia
+Tomocichla, is endemic to the Chortis terrane following
colonization from the Maya terrane and was separated from
Table 1 Data partitions, number of sites (bp) and informativeness [parsimony informative (PI) sites; PI sites with indels in
parentheses], and selected models of evolution for the seven molecular markers (cyt b,ND4,COI,16S,RAG1,RAG2,S7i1). Codingmit
and codingnuc partitions include coding mitochondrial and coding nuclear markers, respectively. Nocodingmit and nocodingnuc
partitions include non-coding markers. Pos1 +2 and pos3 denote first and second versus third codon positions.
Seven partitions Sites PI sites % PI Model Four partitions Sites Model Five partitions Sites Model
cyt b1137 499 44 GTR+I+G Codingmit 2401 GTR+I+G Codingmit pos1 + 2 1601 HKY+I+G
ND4 676 319 47 GTR+I+G Codingmit pos3 800 GTR+I+G
COI 590 218 37 GTR+I+G
16S 586 130 (138) 22 (24) GTR+I+G Nocodingmit 586 SYM+G Nocodingmit 586 SYM+G
RAG1 1475 81 5 K80+I+G Codingnuc 2346 SYM+G Codingnuc 2346 SYM+G
RAG2 871 80 9 K80+G
S7i1 579 139 (172) 24 (30) HKY+G Nocodingnuc 579 HKY+G Nocodingnuc 579 HKY+G
TOTAL 5914 1466 (1507) 25 (25) 5912 5912
cyt b, cytochrome b;ND4, NADH dehydrogenase subunit 4; COI, cytochrome coxidase subunit I; S7i1, S7 intron 1.
Journal of Biogeography 40, 579–593
ª2012 Blackwell Publishing Ltd
583
Biogeography of Mesoamerican Cichlidae
Figure 1 The 50% majority-rule consensus tree of the Mesoamerican Cichlidae plus outgroups obtained from the Bayesian analysis of
the combined data set (mitochondrial DNA: cytochrome b, NADH dehydrogenase subunit 4, cytochrome coxidase subunit I, 16S;
nuclear DNA: RAG1,RAG2, S7 intron 1), analysed as five data partitions (see Table 1). Support values (PP) are shown to the right of
the nodes. Higher level taxonomy is indicated for four clades. The three nodes used for molecular clock calibration in Fig. 2 are shown
with numbers in black circles (see text). The scale bar represents the average number of substitutions per site.
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its sister clade at 26.3 Ma (30.1–22.9 Ma, 95% HPD; Fig. 2).
The fourth subclade, Astatheros +Rocio, is ancestrally
endemic to the Maya terrane, with Rocio and the basal As-
tatheros macracanthus still endemic there (Fig. 2). The basal
divergence within the herichthyine cichlids, separating the
Astatheros +Rocio subclade from the rest of the clade,
occurred at 28.9 Ma (32.7–25.2 Ma, 95% HPD; Fig. 2).
The second Middle American clade (the amphilophines)
has a much more complex internal phylogeny, marked by
short internal branches. The clade is ancestrally endemic to
the Chortis terrane (the San Juan IP; Figs 2, 3a,b & 4a,b)
where it still has the highest diversity at the terrane’s south-
ern edge, i.e. at its contact zone with lower Central America.
There are only three strongly supported suprageneric clades
within the amphilophines: (1) Parachromis plus one lineage of
Cryptoheros; (2) Hypsophrys plus another lineage of Cryptoh-
eros; and (3) the morphologically disparate clade includ-
ing Petenia, the ‘Heros’ urophthalmus group and ‘Heros’
calobrensis.
The amphilophine cichlids have a much wider distribution
in Central America than the herichthyines, and several of the
amphilophine lineages suggest a quite early colonization of
eastern Panama via developing lower Central America (the
Talamanca terrane of oceanic crust; Figs 3a & 4b). The ‘Her-
os’ sieboldii lineage reached eastern Panama already before
18.2 Ma (21.1–15.4 Ma, 95% HPD), ‘Heros’ calobrensis
between 16.5 and 16.3 Ma and the Cryptoheros panamensis
lineage between 15.0 and 12.7 Ma (17.4–9.8 Ma, 95% HPD;
Fig. 2). The colonization dates are largely congruent and
suggest that favourable conditions (continuous land or suc-
cessive stepping stones) were present between 19 and 13 Ma
(Figs 2 & 4b). The vicariant events in the above taxa
separating species in eastern Panama from those in the Ta-
lamanca terrane at the Culebra Strait (i.e. the Panama Canal
Zone) occurred between 12.9 and 10.3 Ma (12.9, 12.7 and
10.3 Ma, respectively; 15.6–6.6 Ma, 95% HPD; Figs 2 & 4c).
These dates thus set the formation of the Tuira IP of eastern
Panama (Fig. 4c).
IPs started to form within the Talamanca terrane (Chiriqui,
Bocas), based on the cichlid dating, during the same time
(Figs 2 & 4c) but took slightly longer to complete due to
repeated colonizations from the Chortis block (Figs 2 &
4c,d): Chiriqui between 14.2 and 6.3 Ma (14.2, 11.8, 10.3,
8.7, 6.3 Ma, respectively; 18.4–4.3 Ma, 95% HPD), Bocas
between 11.5 and 4.3 Ma (11.5, 5.6, 4.3 Ma, respectively;
15.6–2.0 Ma, 95% HPD). On the contrary, the separation of
the Chortis terrane into the San Juan and Chiapas–Nicaragua
IPs is still not complete at the species level (Figs 2 & 4d).
Two additional colonizations of eastern Panama directly
from South America are more recent, having occurred
through two different corridors (Figs 2 & 4d) before 6.1 Ma
(10.2–2.8 Ma, 95% HPD) in Caquetaia and between 3.6 and
2.3 Ma (5.8–3.9 Ma, 95% HPD) in ‘Heros’ atromaculatus,
respectively (i.e. in the period of the GAI).
All colonization and vicariant events from Fig. 2 are
summarized in Appendix S3.
DISCUSSION
Cichlid colonization of Mesoamerica
Cichlids have undoubtedly reached the Greater Antilles and
Middle America by dispersal, but the question is from where,
how and when. Rosen (1975) and later Chakrabarty (2006)
and Chakrabarty & Albert (2011) postulated colonization of
the Greater Antilles through Middle America from the Maya
terrane (roughly corresponding to present-day Yucatan).
Iturralde-Vinent & MacPhee (1999) presented an alternative
reconstruction postulating overland or island-hopping
dispersal to the Greater Antilles from South America. They
termed their hypothetical land bridge the ‘GAARlandia’,
an acronym combining the Greater Antilles with the now
submerged terranes of the Aves Ridge.
Iturralde-Vinent & MacPhee (1999) established (on the
basis of evidence by Nagle, 1972; Bouysse et al., 1985;
Holcombe & Edgar, 1990) that before the Miocene, most of
the topographic highs on the Aves Ridge, linked to the
Aruba/Tobago belt, may have been subaerial and would have
formed a string of emergent lands along a north–south axis
(Fig. 3a). The Aves Ridge and the north-western South
American microcontinent were extensively and coterminously
uplifted and connected with the Greater Antilles around the
Eocene–Oligocene boundary (35–33 Ma), as suggested by the
absence of marine sediments of this age in these areas
(Fig. 3a). The Late Oligocene (29–24 Ma), in addition, expe-
rienced a significant drop in the global sea levels of more
than 150 m (Haq et al., 1987; Figs 2 & 3a). After the highest
emergence and lowest sea levels in the Oligocene period, the
Aves Ridge rapidly subsided (during the Late Oligocene, 27–
25 Ma), was inundated by rising sea levels (before the start
of the Miocene around 23.5 Ma; Fig. 3b), and was deeply
submerged by the Middle Miocene (16–14 Ma; Bock, 1972;
Nagle, 1972; Bouysse et al., 1985; Fig. 3c). Our molecular
clock-dated biogeographical analyses (Fig. 2) with the palae-
ogeographical reconstructions of Mesoamerica (Fig. 3)
strongly agree and support cichlid colonization of the
Greater Antilles, together with Middle America, through the
GAARlandia land bridge from South America (Fig. 3a). Our
results demonstrate that the heroine cichlids colonized Meso-
america during a relatively narrow time window between 35
and 29 Ma in the Oligocene (Figs 2 & 3a). This is in very
good agreement with both the postulated maximum emer-
gence of GAARlandia dated at 35–33 Ma and with the signif-
icant drop in sea levels dated at 29–24 Ma (Figs 2 & 3a).
The termination of GAARlandia, based on our results,
occurred between 26.6 and 24.2 Ma (the vicariances of the
Antillean Nandopsis, and the ‘Heros’ festae group from their
sister groups, respectively; see Fig. 2). This date agrees with
the start of rising sea levels and the postulated fragmentation
of GAARlandia around 23.5 Ma (Figs 2 & 3b). The coloniza-
tion and vicariance dates thus agree with the GAARlandia
land bridge hypothesis. In addition, the close relationship
between the Greater Antillean Nandopsis and the Middle
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American amphilophines, with a reconstructed common
ancestral area for colonization of Middle America and the
Greater Antilles (dated at 29.6–28.0 Ma; Fig. 2) provide
additional evidence that the route of colonization passed
through the GAARlandia land bridge. Also the biogeography
of cichlid fishes within Middle America (see below) supports
our reconstructed colonization. Our biogeographical recon-
struction for the colonization of Mesoamerica thus reverses
the proposed cichlid colonization route of the Greater Antil-
les through Middle America (over Yucatan), proposed by
Chakrabarty (2006) and Chakrabarty & Albert (2011).
Based on available evidence, cichlids probably had to cross
at least narrow gaps between emergent lands during their
colonization. During their colonization in the Oligocene, the
closest land connection of Middle America with South Amer-
ica was via the Greater Antilles (Fig. 3a). However, there was
probably never a direct overland connection between Middle
America and the Greater Antilles during this time (Iturralde-
Vinent, 1969; Iturralde-Vinent & MacPhee, 1999). Further-
more, data from offshore seismic lines indicate that the
Yucatan Channel is an ancient feature that had existed long
before the Late Eocene (probably since the Maastrichtian;
Case & Stehli, 1975; Mascle et al., 1985; Rosencrantz, 1990).
Therefore, for most of the Cenozoic, if not longer, the Cuban
terranes and the Maya terrane have been separated by a nar-
row water barrier (Iturralde-Vinent & MacPhee, 1999). This
could explain why the secondary freshwater cichlids and poe-
ciliids became the dominant fish groups of Mesoamerica
instead of the primary freshwater Ostariophysi that dominate
South America. The colonization through this narrow barrier
is, however, much more likely than direct colonization
through a wide ocean from South America.
Early diversification of the heroine cichlids in Middle
America
The two main Middle American heroine cichlid clades (the
herichthyines and the amphilophines) have different ancestral
areas in Middle America (Figs 2, 3a & 4a). They thus possibly
took different routes from the Greater Antilles, or more likely,
their history was subject to extensive extinctions (Figs. 2 &
3a). The ancestral area of the herichthyine cichlids is the Maya
terrane of the North American Plate (the present-day Usumac-
inta IP), while that of the amphilophine cichlids is the San
Juan IP of the Chortis terrane of the Caribbean Plate.
The older age of the northern Middle American ancestral
areas, compared with the rest of Middle America to the
south, is strongly supported by geological and palaeogeo-
graphical reconstructions. During the period in question,
emergent land existed only in northern Middle America
(Figs 3a & 4a). The northern part of the geologically complex
Middle America (the Maya and Chortis terranes) is old and
made of continental crust, whereas the southern portion –
lower Central America –is a geologically newer volcanic arc
made of oceanic crust (Coates, 1997; Fig. 4a,b). The line sepa-
rating the North American Maya and the Caribbean Chortis
blocks runs through the Cayman Trough and Polochic–Mota-
gua fault system which lies under the sea between Cuba and
Guatemala (Amatique Bay), then runs through Guatemala
and continues as the Sierra de Chiapas, dividing Caribbean
and Pacific drainages in southernmost Mexico and the Gra-
ben systems of Honduras (Guzma
´n-Speziale, 2001). Based on
our results the Maya and the Chortis blocks were probably
already separated (by the Motagua Fault and Amatique Bay)
at the time of cichlid colonization of Middle America (Figs 2,
3a & 4a), but secondary dispersals and vicariances did occur
(Figs. 2 & 4a–d). Lower Central America (i.e. the Panama
microplate, part of the Caribbean Plate) was separated from
the Chortis Block throughout most of the Cenozoic by a mar-
ine corridor known as the Nicaraguan Trough (currently
occupied by the Great Lakes of Nicaragua; Coates & Obando,
1996; Fig. 4b–d). The Talamanca terrane south of this sea gap
thus formed a separate (island) unit throughout most of the
Cenozoic evolution of Middle America.
Cichlid colonization of lower Central America is of
Miocene age (approximately 19–13 Ma), i.e. much younger
Figure 2 beast phylogeny of the Mesoamerican Cichlidae (plus outgroup taxa) showing molecular clock date estimates and
biogeographical reconstruction using Bayesian dispersal–vicariance (S-DIVA) analysis. The tree is a chronogram based on the same data
set as in Fig. 1. The inset figure shows the location of the 14 geographical areas (corresponding to ichthyological provinces) used for
the S-DIVA analysis. The letter codes for each area are as follows: A, Tamesı
´–Pa
´nuco; B, Usumacinta; C, San Juan; D, Bocas; E, Chagres;
F, Tuyra; G, Santa Maria; H, Chiriqui; I, Chiapas–Nicaragua; J, Balsas; K, north-western Pacific; L, trans-Andean South America (Choco
–Tumbes–Magdalena); M, Greater Antilles; O, cis-Andean South America (‘cis’ and ‘trans’ refer to areas of South America east, and
west and north of the Andes, respectively). Mean age estimates (in Ma) are shown to the right of all nodes. Horizontal bars at nodes
represent 95% highest posterior density (HPD) for age estimates. The distribution of each species, as described by its presence in the
ichthyological provinces (IPs; inset figure), is given in front of the generic names (for corresponding species see Fig. 1 & Appendix S2);
the genera are colour-coded according to their reconstructed ancestral distributions. Ancestral distribution derived from S-DIVA is
indicated for all nodes by the same colour as used for the IPs in the inset figure (pie charts for all internal nodes representing the
probabilities for each alternative ancestral area are shown in Appendix S2). The time chart showing duration of epochs throughout the
Cenozoic is accompanied by the global sea level curve from Haq et al. (1987). The vertical grey bars spanning 32–24 Ma (dark grey)
and 35–19 Ma (light grey) represent the mean and the 95% HPD reconstructed duration of GAARlandia, respectively. The rainbow
colours in the time chart summarize first colonization events of new areas within Mesoamerica: during the Oligocene cichlids have
colonized two areas of Middle America (B and C) together with the Greater Antilles (M) from South America through GAARlandia.
Following a 10-Myr evolutionary stasis (shown by three horizontal stippled boxes on the tree branches) in the San Juan IP (C) cichlids
have then colonized all areas of Middle America between c. 19 and 13 Ma.
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Biogeography of Mesoamerican Cichlidae
(by approximately 10–15 Myr) than the initial Oligocene
colonization of Middle America (Figs 2, 3c & 4b). Only the
amphilophine cichlids colonized lower Central America all
the way to eastern Panama (Figs 1, 2, 3c & 4b). They appear
to have experienced either a stasis in diversification or extensive
extinctions following their colonization of Middle America (see
above) because they have a long basal branch between their
origin at 28 Ma and their first recorded diversification at
18 Ma (Fig. 2). Long branches of similar duration without
diversification are also notable in both the other lineages
ancestrally present in the Chortis Block (Astatheros and
Tomocichla; see Fig. 2). The diversification of the amphilo-
phine cichlids was then rapid, and all their major subclades
and genera originated between 19 and 13 Ma. Our analyses
indicate that during this time interval they managed to cross
the gap between the Chortis and Talamanca terranes and
Figure 3 Historical biogeography of
heroine cichlids in Mesoamerica from the
Early Oligocene to the Early Miocene.
Palaeogeographical maps of Mesoamerica
are based on Iturralde-Vinent & MacPhee
(1999). (a) Conditions during the period of
maximum Cenozoic land exposure (Late
Eocene/Early Oligocene) occasioned by a
very low eustatic sea level (see Fig. 2) and
widespread regional uplift (the depicted
shorelines and the contact point with South
America are conjectural). Arrows show
cichlid dispersal events. Colours of arrows
correspond to geographical areas in Fig. 2.
Date estimates of each colonization event
based on Fig. 2 are also shown. (b)
Conditions during general subsidence and
higher sea levels (see Fig. 2) during the
Early to Late Oligocene transition that
greatly diminished land area within the
Caribbean Sea, sundering the land bridge
connection between the Greater Antilles and
north-western South America. Two-way
arrows show vicariant events based on
Fig. 2. (c) Isolated tectonic blocks and
terranes of the Antillean Arch separated by
deep-water gaps (Early Miocene). Lower
Central America and South America are not
in contact across the Panamanian region.
Arrows show colonization of lower Central
America from northern Middle America
(see Fig. 4b,c,d).
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disperse to eastern Panama (Figs 2, 3c & 4b). This possibility
has recently been supported by the presence of North Ameri-
can mammals in a mid-Miocene formation in central Pan-
ama, which suggests that emergent land stretched between
North America and Panama between 19.5 and 14 Ma
(Fig. 4b; Kirby & MacFadden, 2005; MacFadden, 2006; Kirby
et al., 2008).
The water gaps on both sides of the Talamanca terrane
(the Nicaraguan Trough and the Culebra Strait) reappeared,
based on our data, around 15–10 Ma (Figs 2 & 4c; see
Results), and lower Central America remained isolated from
nuclear Central America until the end of the Miocene as a
result of collision between the lower Central American por-
tion of the Caribbean Plate with South America (Fig. 4c,d).
This isolation of the Talamanca terrane from eastern Panama
led to the origin of the Tuira IP, and rising elevation of the
terrane to the formation of the Bocas and Chiriqui IPs
(Fig. 4c,d).
Figure 4 Historical biogeography of
heroine cichlids in Middle America from
the Oligocene to the Pliocene (see Fig. 2
and Appendix S3). The reconstructions
show four time frames relating the collision
of Middle America with South America (see
Discussion): (a) Oligocene, (b) Early/Middle
Miocene, (c) Middle Miocene, (d) Late
Miocene/Pliocene. The two main centres of
cichlid evolution in Middle America are the
Usumacinta (Maya terrane; area B) and San
Juan (Chortis terrane; area C) ichthyological
provinces. Colonization routes are depicted
as one-way arrows and vicariant events by
two-way arrows. Each arrow may represent
identical events in several taxa. The
separation of the western part of the San
Juan IP into the separate Chiapas–
Nicaragua province is not completely
finalized to this day in the case of the
Cichlidae (dotted line).
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Biogeography of Mesoamerican Cichlidae
Direct colonization of Middle America by heroine cichlids
from South America happened only with the closure of the
Atrato Seaway towards the end of Miocene, i.e. during the
GAI, but it was limited just to eastern Panama. Only two
cichlid lineages (Caquetaia and the ‘Heros’ atromaculatus
group) entered eastern Panama through two different corri-
dors from trans-Andean north-western South America at 6.1
and 2.3 Ma, respectively (Figs 2 & 4d).
Support for the GAARlandia hypothesis
A biotic exchange between northern South America and the
Greater Antilles through GAARlandia has been invoked for
many terrestrial and freshwater taxa (Ali, 2012), including
mammals such as megalonychid sloths (MacPhee & Iturral-
de-Vinent, 2000; White & MacPhee, 2001; Da
´valos, 2004),
hystricognath rodents (Woods et al., 2001; Da
´valos, 2004),
some bats (Da
´valos, 2004) and primates (Horovitz &
MacPhee, 1999; Da
´valos, 2004), and also frogs (Crawford &
Smith, 2005; Moen & Wiens, 2009; Alonso et al., 2012),
spiders (Binford et al., 2008; Crews & Gillespie, 2010), plants
(Fritsch, 2003; van Ee et al., 2008) and freshwater fishes
(Murphy et al., 1999; Perdices et al., 2005; Concheiro Pe
´rez
et al., 2007; Hrbek et al., 2007; Hulsey et al., 2011).
The GAARlandia land bridge hypothesis predicts a peak
in the arrival of terrestrial (and freshwater) taxa into
Mesoamerica at the Eocene–Oligocene boundary, around 35
–33 Ma. Only a few of the above studies have employed
well-calibrated molecular clock-dated phylogenetic analyses
with historical biogeographical methods to distinguish
between the spatial and temporal alternatives pertaining to
the colonization of Mesoamerica (Cretaceous–Palaeocene
proto-Antilles land bridge; mid-Cenozoic GAARlandia land
bridge; direct, probably multiple and independent overwater
colonization). Older reviews of amphibians and reptiles
based mostly on immunological distances claim that this
peak is absent (Hedges, 2006; but based on data in Hedges,
1996), while reviews of mammals admitted that the evidence
from mammalian phylogenies so far does not add to the
argument of dispersal versus vicariance because of the lack
of proper data and methodology (Da
´valos, 2004; again
based on much older studies). More recent studies that have
been properly dated with molecular clocks together with
historical biogeographical reconstructions, however, tend to
support this peak and show similar results to our study
(Crawford & Smith, 2005; Hrbek et al., 2007; Alonso et al.,
2012). Hrbek et al. (2007) found that in the freshwater fish
group Poeciliidae only one species is referable to coloniza-
tion during the Cretaceous (the Guatemalan endemic
Xenodexia ctenolepis), while the main colonization of Meso-
america occurred in the Eocene/Oligocene, most likely
through GAARlandia. This colonization from South Amer-
ica shows a basal split between taxa found in the Greater
Antilles (namely Cuba) and in Middle America as in the
case of the heroine cichlids (Nandopsis versus amphilo-
phines). The same scenario and palaeobiogeography were
also reconstructed in the second cited study focusing on the
Mesoamerican direct-developing frogs (Leptodactylidae:
Eleutherodactylus s.l.; Crawford & Smith, 2005). One lineage,
Craugastor, colonized Middle America from the ancestral
area in South America through the proto-Antilles during
the Cretaceous, with no descendants left in the Greater
Antilles (as in the Poeciliidae). The second lineage shows a
sister-group relationship between the Greater Antillean
(Syrrhophus) and Middle American (Euhyas) genera, with a
very good accord (35–26 Ma) to the dating of the Oligo-
cene GAARlandia land bridge.
Whether GAARlandia indeed played a role similar to the
GAI still remains to be decided, but if it did there must have
been a similar massive biotic interchange focused into a rela-
tively narrow time window. Both the Antilles and Middle
America are conspicuous in their limited higher-order taxo-
nomic composition, as is the case in all large islands that
have had limited historical connections to their closest main-
land (e.g. Madagascar, Sulawesi, the Philippines). With more
studies using a proper methodological framework we should
be able to see the proportion of taxa showing a coterminous
colonization and reconstructed palaeogeography consistent
with GAARlandia, as opposed to overwater dispersal with
random time distribution.
Our study contributes to the evidence that GAARlandia
was an important geographical and faunal element during the
Oligocene. We demonstrate that there are closely related (even
sister) taxa inhabiting the Greater Antilles and Middle Amer-
ica, and that the often depauperate Antillean taxa are in sev-
eral cases remains of a wider colonization of Mesoamerica.
The Antilles have probably experienced more extinctions than
Middle America because of their smaller size, and this could
have clouded our understanding of Mesoamerican biogeogra-
phy and led us to underestimate the role that the Antilles
played in the colonization of Mesoamerica.
ACKNOWLEDGEMENTS
We are indebted to the late Gustavo A. Concheiro Pe
´rez
(Smithsonian Tropical Research Institute) for sharing collec-
tion records, and to three anonymous referees and the editor
for suggestions that significantly improved previous versions
of the manuscript. Financial support was provided by grants
from the SYNTHESYS programme for Access to the Museo
Nacional de Ciencias Naturales (Madrid, Spain) within the
European Community programme ‘Improving the Human
Research Potential and the Socio–Economic Knowledge
Base’, and by the GAC
ˇR 206/08/P003 and MSM6007665801
grants to O.R
ˇ. and J.Z.
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SUPPORTING INFORMATION
Additional Supporting Information may be found in the
online version of this article:
Appendix S1 Sampling table and main sources of data.
Appendix S2 Results of the statistical dispersal–vicariance
analysis (S-DIVA).
Appendix S3 Summary of colonization and vicariance
events based on the analysis in Fig. 2.
As a service to our authors and readers, this journal pro-
vides 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 supporting information (other
than missing files) should be addressed to the authors.
BIOSKETCH
Oldr
ˇich R
ˇı
´c
ˇan is researcher and lecturer in zoology inter-
ested in the evolution of life-history traits and spatial distri-
bution of biodiversity, particularly in freshwater fishes of the
Neotropical region.
The main research interests of all the authors are the evolu-
tion, phylogenetics and biogeography of animals.
Author contributions: O.R
ˇ. and J.Z. conceived the study.
O.R
ˇ. provided the additional material and artwork used in
this study, performed most of the analyses and led the
writing. L.P. conducted the molecular laboratory work and
thealignmentofthesequencedata.I.D.andR.Z.provided
laboratory space and financial and technical support in the
early stages of the work. All authors contributed to the
writing of the manuscript.
Editor: Alistair Crame
Journal of Biogeography 40, 579–593
ª2012 Blackwell Publishing Ltd
593
Biogeography of Mesoamerican Cichlidae
