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Molecular Phylogenetics and Evolution 47 (2008) 251–260
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A complete species-level molecular phylogeny for the ‘‘Eurasian”
starlings (Sturnidae: Sturnus, Acridotheres, and allies):
Recent diversification in a highly social and dispersive avian group
Irby J. Lovette a,*, Brynn V. McCleery a, Amanda L. Talaba a, Dustin R. Rubenstein a,b,c
aFuller Evolutionary Biology Program, Laboratory of Ornithology, Cornell University, Ithaca, NY 14950, USAb
Department of Neurobiology and Behavior, Cornell University, Ithaca, NY 14850, USAc
Department of Integrative Biology and Museum of Vertebrate Zoology, University of California, Berkeley, CA 94720, USA
Received 2 August 2007; revised 17 January 2008; accepted 22 January 2008
Available online 31 January 2008

Abstract
We generated the first complete phylogeny of extant taxa in a well-defined clade of 26 starling species that is collectively distributed
across Eurasia, and which has one species endemic to sub-Saharan Africa. Two species in this group—the European starling Sturnus
vulgaris and the common Myna Acridotheres tristis—now occur on continents and islands around the world following human-mediatedintroductions, and the entire clade is generally notable for being highly social and dispersive, as most of its species breed colonially or
move in large flocks as they track ephemeral insect or plant resources, and for associating with humans in urban or agricultural landscapes. Our reconstructions were based on substantial mtDNA (4 kb) and nuclear intron (4 loci, 3 kb total) sequences from 16 species,
augmented by mtDNA NDII gene sequences (1 kb) for the remaining 10 taxa for which DNAs were available only from museum skinsamples. The resulting mitochondrial gene tree embedded within a multilocus framework shows that the well-studied taxa S. vulgaris/
unicolor are the sister lineage to the remaining members of the radiation, from which other relatively early lineages gave rise to forms
that are now nomadic or locally migrant in Africa (Creatophora) and western Asia (Pastor). The remaining taxa form a clade with acomplicated biogeographic history primarily in central and eastern Asia; this group contains a range of sedentary to highly migratorytaxa, as well as widely distributed species and single-island endemics such as the highly endangered Bali myna (Leucopsar). Several
groups of species in the genus Acridotheres have low magnitudes of within-group divergence and likely diversified via their respective
colonization of islands. The taxonomy of this entire group has remained highly volatile over the past century; we propose dividing these26 species among 11 reciprocally monophyletic genera (Acridotheres, Poliopsar, Temenuchus, Sturnornis, Leucopsar, Gracupica, Agropsar,
Pastor, Creatophora, and Sturnus).
! 2008 Elsevier Inc. All rights reserved.
Keywords: Sturnidae; Starling; Sturnus vulgaris; Phylogeny; Taxonomy

1. Introduction
Starlings have radiated impressively throughout muchof Eurasia, Africa, and the eastern Pacific. Recent phylogenetic work has defined the taxonomic and geographic distributions of the six major clades within the starlingfamily Sturnidae (Zuccon et al., 2006; Lovette and Ruben*

Corresponding author. Fax: +1 607 254 2486.
E-mail address: IJL2@cornell.edu (I.J. Lovette).

1055-7903/$ - see front matter ! 2008 Elsevier Inc. All rights reserved.
doi:10.1016/j.ympev.2008.01.020

stein, 2007). One of these sub-groups, which we informallyterm the ‘‘Eurasian” starlings, is particularly notable forcomprising species that are nearly all disturbed habitat specialists, many of which now occur as human commensals inurban or agricultural landscapes (Feare and Craig, 1999).
Nearly all of the 26 currently recognized extant species(Dickinson, 2003) in this Eurasian clade are likewise atleast partially nomadic or migratory, and most breed colonially and/or flock during the non-breeding season (Feareand Craig, 1999). This combination of high dispersal

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potential and the simultaneous movements of groups ofindividuals has likely facilitated the natural geographic
expansion of this group across Eurasia and (by one species)
throughout much of Africa, giving this group a greater
geographic breadth than that of any equivalent sub-clade
within the Sturnidae. The distribution of the Eurasian starling clade has been expanded further by the extensive
human-mediated introduction of two species (the European starling Sturnis vulgaris and the common myna Acridotheres tristis) to all non-polar continents and to hundreds
of islands in both hemispheres (Feare and Craig, 1999).
This clade of starlings is further important because Sturnus
vulgaris has long been the subject of extensive ecological,
physiological, and behavioral research (Feare, 1984), andplacing such studies in a comparative context requires phylogenetic information on how vulgaris is related to alliedtaxa.
The taxonomy of this group of starlings has been highly
volatile over the past century, particularly at the genuslevel (Table 1), owing largely to the lack of known morphological synapomorphies within the group. Several workers
with extensive systematic expertise with starlings have commented on the confusing relationships among these taxa
(Amadon, 1943; Beecher 1978; Feare and Craig, 1999).
Beecher (1978) generated a dendogram of starling genera
based on his studies of jaw musculature, in which the genera in our Eurasian clade cluster together. A terminal

branch in the DNA–DNA hybridization studies of Sibley
and Ahlquist (1984, 1990) similarly groups together thefour Eurasian species they investigated. More recently,
one to three species in the Eurasian clade have been
included in a number of molecular phylogenetic studies
of deeper songbird relationships (Barker et al., 2004; Voelker and Spellman, 2004; Spicer and Dunipace, 2004; Ericson and Johansson, 2003; Cibois and Cracraft, 2004;
Alström et al., 2006), and one recent study (Zuccon
et al., 2006) included eight. Kang et al. (1994) used cytochrome b sequences to examine relationships among threeclosely allied Acridotheres species.
The most extensive previous phylogenetic sampling of
the Eurasian starlings was in a recent mtDNA and intron
based study by our group (Lovette and Rubenstein, 2007)
that included 20 Eurasian clade species along with robust
sampling of all other Sturnidae clades and genera. Our previous work was directed largely towards defining the various major clades within the deeper Sturnidae radiation,and it provided unambiguous support for the in-group considered here, as the basal internode leading to the Eurasianclade was long and universally highly supported acrossdatasets and analysis methods (Lovette and Rubenstein,2007). Other relevant findings from our previous study
include: (i) the likely position of Sturnus vulgaris/unicolor
as the basal lineage within the Eurasian starling clade; (ii)the inclusion of the African species Creatophora cinerea

Table 1
Taxonomic recommendations for ‘‘Eurasian” starlings based on their molecular phylogenetic affinities, and comparisons to five recent taxonomic
treatments of this group
Proposed here
Genus

SpeciesAcridotheres
Acridotheres
Acridotheres
Acridotheres
Acridotheres
Acridotheres
Acridotheres
Acridotheres
Acridotheres
Acridotheres
Poliopsar
Poliopsar
Temenuchus
Temenuchus
Temenuchus
Temenuchus
Sturnornis
Leucopsar
Gracupica
Gracupica
Agropsar
Agropsar
PastorCreatophora
Sturnus
Sturnusgrandisacristatellusa
albocinctusa
javanicusb
cinereusb
fuscusb
melanopterus
burmannicus
ginginianus
tristissericeus
cineraceus
sinensismalabaricus
erthropygius
pagodarum
albofrontata
rothschildi
nigricollis
contrasturninus
philippensis
roseuscinereavulgarisc
unicolorc

a,b,c

Amadon (1962)

Wolters (1982)

Sibley and Monroe (1990)

Feare and Craig (1999)

Dickinson, (2003)———
ssp. fuscus
Sturnus—
Sturnus———
Sturnus
Sturnus
Sturnus
Sturnus
Sturnus
Sturnus
Sturnus—
Sturnus
Sturnus
Sturnus
Sturnus
Sturnus———

Aethiopsar
Aethiopsar
Aethiopsar
ssp. fuscus
Aethiopsar
Aethiopsar
Leucopsar
Leucopsar——
Sturnopastor
Sturnopastor
Sturnia———
Temenuchus——
Sturnopastor
—
—
—
—
—
—

—
—
—
—
ssp. fuscus—
Sturnus———
Sturnus
Sturnus
Sturnus
Sturnus
Sturnus
Sturnus
Sturnus—
Sturnus
Sturnus
Sturnus
Sturnus
Sturnus
—
—
—

—
—
—
—
—
—
—
Sturnus——
Sturnus
Sturnus
Sturnia
Sturnia
Sturnia—————
Sturnia
Sturnia
—
—
—
—

—
—
—
—
—
—
—
—
—
—
Sturnus
Sturnus
Sturnus
Sturnus
Sturnus
Sturnus
Sturnus—
Sturnus
Sturnus
Sturnus
Sturnus
Sturnus———

Superscripts denote three clades within which species are separated by very low levels of genetic differentiation.

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as the sole non-Eurasian member of the clade, which isconsistent with previous suggestions based on its morphological and behavioral similarities to the remaining species
in this group (Amadon, 1943, 1956; Feare and Craig, 1999;
Fry et al., 2000); and (iii) a lack of close correspondence
between the DNA based reconstructions and any previous
genus-level classification of these taxa.
Here, we reconstruct relationships among all 26 extant
species in the Eurasian starling group. High-quality genetic
materials were available from 16 species, and for these taxa
we have substantial mitochondrial DNA and nuclear
intron information. For 10 species, however, we have only
mitochondrial sequences derived from museum skin samples. The combined analysis of these nested datasets allows
us to place the mtDNA-only gene tree within the morerobust phylogenetic framework provided by the more
extensive, multilocus markers, and we discuss the opportunities and caveats associated with this combination ofmarkers. The resulting reconstructions are the first complete phylogenies for this clade of extant starlings; they
provide a formal phylogenetic basis for revised genus-leveltaxonomy and allow us to examine the evolution of the jaw
musculature traits that have formed the primary basis forthe existing taxonomy of this group.
2. Methods
2.1. Taxon sampling and laboratory methods
For this study, we sampled at least one representative of
all extant ‘‘Eurasian” Sturnidae taxa that have been considered full species in any of five recent classifications of
the family (Table 1). Twenty of these taxa were included
in our previous study of Sturnidae phylogenetics (Lovette

and Rubenstein, 2007) and the corresponding sample
sources and sequence data are described in detail there.
Samples new to this analysis are described in Table 2. In
all cases, these new samples were derived from toe-pad tissues taken from existing museum skin materials. Toe-padskin samples were taken by shaving a narrow band of skinfrom a single toe (usually the hallux), using a new, sterile
scalpel. These skin samples were placed dry into sterile
1.5 mL tubes, sealed, transferred to our degraded-DNA
laboratory, and stored at room temperature until DNA
extraction. We also attempted to generate sequences fromthe extinct Reunion Island taxon Fregilupus varius, which
has some morphological similarities to the Sturnus starlings(Amadon, 1956), but which has generally uncertain affinities and may not be a starling (Miller 1941; Berger,
1957); we were unable to amplify PCR products using Sturnidae-specific primers from toe-pad skin samples of a Fregilupus specimen collected some time before 1840 that is
now in the collection of the Harvard University Museumof Comparative Zoology. We also did not include Necropsar leguati, a taxon described as a starling but now known
to be based on a fraudulently labeled specimen (Olsonet al., 2003).
From most of the samples included in our previous
study, we have sequences spanning the NDII, COI, COII,
ATPase8, and ATPase6 mitochondrial genes (total length4118 bp), and sequences from four introns totaling nearly
3 kb, including indels after alignment: rhodopsin intron 1(Primmer et al., 2002), intron 5 of transforming growth factor b-2 intron 5 (Primmer et al., 2002), and the closely
linked b-fibrinogen introns 5 (F.K. Barker, personal communication) and 7 (Prychitko and Moore, 1997). From
some additional taxa included previously, and from all
samples new to the present analysis, we obtained only

Table 2
Samples included in this study that have not been reported previously and GenBank accession numbers for the associated NDII sequences
Taxon

Sourcea

Specimen

Yearb

Locality

GenBank No.

Acridotheres albocinctus
Acridotheres albocinctus
Acridotheres cinereus
Acridotheres cinereus
Acridotheres javanicus
Acridotheres javanicus
Acridotheres melanopterus
Acridotheres melanopterus
Acridotheres burmannicus
Acridotheres burmannicus
Acridotheres ginginianus
Sturnornis albofrontata
Temenuchus erythropygius
Temenuchus erythropygius
Agropsar sturninus
Agropsar sturninus

AMNH
AMNH
AMNH
AMNH
AMNH
AMNH
AMNH
AMNH
MVZ
MVZ
AMNH
AMNH
AMNH
AMNH
AMNH
AMNH

347176409723299902299898666486666493417390666360156076156077304811666278666245666249666153666149

1945
1935
1931
1930
1927
1927
1937
1927
1965
1965
1931
1876
1907
1907
1910
1918

Myanmar (Burma)
India: Assam, Dalu
Indonesia: Celebes, LombasangIndonesia: Sulawesi, MakassarIndonesia: Java, Mt. Ohojoeno
Indonesia: Java, Mt. Ohojoeno
Captive bird (New York Zoo)
Indonesia: Java
Vietnam: Saigon
Vietnam: Saigon
Pakistan: Punjab, Lahore
Sri Lanka: ‘‘Powanala”
Nicobar Island
Nicobar Island
Malaysia: Perak
Indonesia: Sumatra, Deli

EU403586
EU403585
EU403589
EU403590
EU403592
EU403593
EU403595
EU403594
EU403587
EU403588
EU403591
EU403596
EU403597
EU403598
EU403600
EU403599

All samples below were derived from toe-pads taken from museum skin specimens.
aMuseum specimen sources: AMNH, American Museum of Natural History, New York, NY, USA; MVZ, Museum of Vertebrate Zoology, Universityof California, Berkeley, USA.
b
Year of specimen collection.

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NDII sequences. Laboratory methods for the generation ofthese sequences are detailed in Lovette and Rubenstein(2007).
All samples new to this study were processed in a laboratory dedicated to degraded-DNA extraction and PCRset up that has physical and air-handling isolation fromour general-use laboratory. Toe-pad extractions and PCR
reactions were assembled and conducted within a laminar-flow clean bench with ISO class V air filtering, andwe frequently sterilized surfaces and equipment in the
degraded-DNA room with intense 254 nm UV irradiationand 10% sodium hypochlorite (chlorine bleach) solutions.
To enhance the probability of detecting any contaminatedreactions, we interspersed negative control reactions at
both the extraction (1 control:1 tissue-containing extraction) and PCR (also 1:1) stages. When sequencing multiple
toe-pad samples from one species, we separated the processing of those different specimens by several weeks tomonths, over which interval many other samples were processed using the same laboratory facility, equipment, primers, and reagents. Degraded-DNA PCR amplifications
targeted short (100–500 bp), overlapping regions of the
NDII gene. We employed numerous primers flanking orwithin the NDII coding region, including many designed
specifically for the taxa included here. Although the detection of preferentially or co-amplified nuclear homologs of
mtDNA sequences is more difficult in studies involving
degraded sample sources than it is when working withhigh-quality samples, we found no evidence of nuclear copies in the chromatogram data or overlapping regions
amplified in separate PCR reactions.
2.2. Phylogenetic analysis
Phylogenetic reconstructions were rooted to the distantougroup Buphagus africanus, but we also included in all
analyses one representative of each of the non-Eurasian
Sturnidae clades and one representative Mimidae species;
these additional taxa were Cinnyricinclus leucogaster, Lamprotornis superbus, Onchognathus morio, Rhabdornis inornatus, and Mimus polyglottos (corresponding sequences allfrom Lovette and Rubenstein, 2007).
To reconstruct phylogenies, we used Bayesian methods
as implemented in MrBayes 3.1 (Huelsenbeck and Ronquist, 2001; Ronquist and Huelsenbeck, 2003). Becauseof the heterogeneous composition of our dataset, we ran
two separate phylogenetic analyses: the first on all data
available and the second restricted to the NDII genesequence subset which we had from all taxa. Bayesian
MCMC chains were sampled every 100 generations, with
two independent sets of three heated and one unheatedchains. All analyses employed the default flat Dirichlet priors. Stationarity was evaluated graphically for all parameters, and by monitoring the convergence of the standard
deviation of split frequencies in the two independent setsof chains. Chains reached stationarity within the first
1 ! 106 generations but were run for 5 ! 106 generations;samples from the initial 4 ! 106 generations were discarded. Congruence between independent runs based on
identical datasets was assessed by comparing parameter
estimates, tree topologies, and posterior probability scoresfor individual branches; in no case did topologies conflictor parameter values substantially differ. The multilocus
analysis included five independent partitions, one for the
mitochondrial sequences and one for each of the four
intron loci; parameters were estimated for each under the
GTR+G+I model. In the NDII-only analysis, all data
were treated as a single partition. An ultrametric NDII
topology was generated in PAUP* 4.0b10 (Swofford,
2002) using the 50% majority-rule consensus topology fromthe NDII-only Bayesian analysis, with maximum likelihood branch lengths calculated using the mean model
parameters from the 1 ! 106 sampled generations (empirical base frequencies; alpha rate parameter = 0.765; proportion of invariable sites = 0.376; relative substitution rates:
A–C = 0.35, A–G = 11.18, A–T = 0.30, C–G = 0.32, C–
T = 4.55, G–T = 1.00).
3. Results
3.1. Phylogenetic hypotheses and the systematics of theEurasian starling clade
We obtained complete NDII gene sequences (1041 bp)
for all samples listed in Table 2. In all cases, NDII
sequences from conspecific samples were identical, or wereseparated by the low level of divergence typical of withinspecies comparisons among different geographic sampling
locations: the largest within-species NDII difference(among two of the three Acridotheres javanicus samples)
was 1.1% uncorrected divergence. All novel sequences havebeen deposited in GenBank (Table 2). In all reconstructions, the Eurasian clade considered here was always recovered with 100% posterior probability support for a long
basal internode separating this in-group from the non-Eurasian representatives of the Sturnidae radiation.
We conducted separate analyses of (i) all data from alltaxa, and (ii) solely the NDII gene sequences that were partof the data subset common to all samples (Fig. 1). Somewhat surprisingly given the much higher information content of the larger dataset, the resulting reconstructions
were almost topologically identical and had nearly equivalent resolution, as indexed by the number and identity ofnodes receiving P95% posterior probability scores. Excluding 14 nodes involving very shallow terminal sister taxa
(usually conspecific replicates) that all received P95%
scores in both trees, only 2 of the remaining 19 nodes with
P95% support in the all data tree received lower support in
the NDII-only reconstruction. Those two variably supported nodes both involve the relationships among Sturnopastor sericeus, Sturnopastor cineraceus, and the
Acridotheres group: in the complete data reconstruction,
sericeus and cineraceus are well-supported sister taxa at
the two relevant nodes, whereas in the NDII-only tree cin-Author's personal copy

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I.J. Lovette et al. / Molecular Phylogenetics and Evolution 47 (2008) 251–260

*
*
*

*
*

*

*

*

*

*

*

*
*

*
*

*
*

*

*

*

*

*
*
*

*

*

*

*

*
*

*
*
*

Acridotheres cristatellus
Acridotheres grandis
Acridotheres albocinctus
Acridotheres albocinctus
Acridotheres javanicus
Acridotheres fuscus
Acridotheres cinereus
Acridotheres cinereus
Acridotheres javanicus
Acridotheres javanicus
Acridotheres melanopterus
Acridotheres melanopterus
Acridotheres burmannicus
Acridotheres burmannicus
Acridotheres ginginianus
Acridotheres ginginianus
Acridotheres tristis
Poliopsar sericeus
Poliopsar cineraceus
Temenuchus malabaricus
Temenuchus erythropygius
Temenuchus erythropygius
Temenuchus pagodarum
Temenuchus sinensis
Temenuchus sinensis
Leucopsar rothschildi
Sturnornis albofrontata
Sturnornis albofrontata
Gracupica nigricollis
Gracupica nigricollis
Gracupica contra
Agropsar philippensis
Agropsar philippensis
Agropsar sturninus
Agropsar sturninus
Pastor roseus

*
*
0.016

*

*

*
0.014

*

*
*
*

0.021

*

*

0.016

0.014
* 0.0250.020 *
0.066
0.073
0.013
0.020

*

*

0.033

0.010

0.013

*

*

0.030

*

0.041

*

*

0.072
0.072

*

0.067

*

0.016

0.068

*

0.012 *

0.020

*

0.064

*

0.019

*

Creatophora cinerea
Creatophora cinerea
Sturnus unicolor
Sturnus vulgaris

0.053

*

*

0.099
0.099

*

0.149

*

Fig. 1. Comparison of Bayesian likelihood reconstructions for Eurasian starling species. Topology at left is based on simultaneous analysis of taxa (thinbranches) represented by mtDNA NDII gene sequences only (1041 bp), and taxa (thick branches) represented by more extensive mtDNA (4116 bp) andintron (2974 bp) sequences. Topology and branch lengths at right are based on analyses of only NDII sequences. Asterisks indicate internodes supportedby P95% posterior probability values. Both analyses were rooted to Buphagus africanus and included representative taxa from other Sturnidae clades (notshown) outside the monophyletic group depicted here.

eraceus is basal (with 60% and 81% posterior probabilities
for the two nodes in this alternative topology) to sericeus + Acridotheres. The only other topological difference
between the reconstructions from the two datasets has Leucopsar sister to Sturnia in the complete data tree, and Leucopsar sister to Temenuchus in the NDII-only tree; in both
trees, however, the relevant internode is very short andweakly supported. In no case, therefore, is a well-supported
relationship in one tree in conflict with an alternative wellsupported topology in the other tree. This high level of congruence may stem in part from the fortuitous distribution ofrobustly sampled taxa across the radiation: taxa that were
included via NDII sequences alone were usually closely
allied to taxa from which we have the full set of sequencesfrom both mitochondrial and nuclear DNA (Fig. 1).

4. Discussion
4.1. Systematics of Eurasian starling genera
Although a few nodes in the phylogeny remain poorly
resolved, the reconstructions presented here are consistentin their strong support for internodes in the deeper part of
the radiation that can be used to define genera. In applying
these phylogenetic hypotheses to inform a revised generictaxonomy for these 26 species of starlings, we follow the
principle that genera correspond to monophyletic groups
given the best available phylogenetic evidence. We also
attempt to follow previous taxonomic designations where
possible (while noting that this group has long suffered froma fairly dramatic nomenclatural inconsistency; Table 1). We

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propose now dividing these 26 taxa among the 11 genera
outlined below.
4.1.1. Acridotheres Vieillot 1816, and Poliopsar Sharpe 1888
The most species of our proposed genera is Acridotheres;
all of its 10 specioses have been assigned to Acridotheres inmost recent taxonomies (Table 2), and this genus is definedby a long and well-supported basal branch (Fig. 1). The
probable sister-group to Acridotheres is comprised of two
species (sericeus and cineraceus) which we assign to Poliopsar. This genus, for which cineraceus is the type, has generally been subsumed into Sturnus (Table 2). Our Poliopsar
includes one migratory and one non-migratory species withseasonally overlapping ranges (Feare and Craig, 1999).
Although the sister-relationship of sericeus and cineraceus
was not recovered in the NDII-only reconstruction, bothof these species were represented in our more robust
mtDNA + intron dataset, and their sister-relationshipwas highly supported in the reconstructions based on thoseadditional sequences and loci (Fig. 1). The monophyly of
the taxa we include in Acridotheres + Poliopsar was also
universally highly supported, and an alternative classification is to merge all 13 species into Acridotheres.
4.1.2. Temenuchus Cabanis 1815, Leucopsar Stresemann
1912, and Sturnornis Legge 1879
A second well-supported group includes taxa we assign
to Temenuchus (4 species), Leucopsar (1 species), and Sturnornis (1 species). Within Temenuchus, the basal split separates the migratory sinensis of SE Asia from pagodarum(which is widespread on the Indian sub-continent) and malabaricus (broadly distributed from India to southeast Asia,
overlapping with both sinensis and pagodarum). The fourth
species in this group, erythropygius, is sister to malabaricus,
and probably stems from a colonization of the remote Andaman/Nicobar archipelago in the eastern Indian Ocean by
an early representative of the malabaricus lineage. The Bali
myna, the sole species in Leucopsar, is restricted to theisland of Bali, where it is critically endangered; this species
is morphologically distinctive owing to its largely whiteplumage, and the phylogenetic results suggest that it has
no close extant allies. The same is true for the monotypic
Sturnornis albofrontata, which is a rare endemic in south–
west Sri Lanka with a complicated nomenclatural history
resulting from a misidentification of the original type specimen (see Mees, 1997). The pattern of relationship among
these three genera was not well-resolved in our trees(Fig. 1), but their collective monophyly was highly supported, and an alternative taxonomic treatment would be
to merge all six species into Temenuchus.
4.1.3. Gracupica Lesson 1831, and Agropsar Oates 1889
A third well-supported group contains two species eachin Gracupica and Agropsar. Feare and Craig (1999) noted
the morphological and behavioral similarities of nigricollis
and contra, and we follow them in placing these two sister
species in Gracupica. The two sister-species in Agropsar—

sturninus and philippensis—have been previously assigned
to various genera, but were separated into Agropsar byWolters (1982). Both of these Agropsar species are long-distance migrants that breed allopatrically in northeastern
Asia. An alternative taxonomic treatment would be tomerge Agropsar into Gracupica.
4.1.4. Creatophora Lesson 1847, and Pastor Temminck 1815
The remaining taxa in the Eurasian radiation each havelong branches that leads to low-diversity terminal groups.
The monotypic genera Creatophora and Pastor have origins at the base of the sister-clade to Sturnus (Fig. 1). Creatophora likely represents an early colonization of subSaharan Africa: this species now occurs in semi-arid savannas throughout much of eastern and southern Africa, often
in association with large mammals (Fry et al., 2000). Creatophora is widespread across Africa where it wanders
nomadically in parts of its range and may be a local
migrant in other areas (Feare and Craig 1999). Given thepossibility that Creatophora colonized Africa early in thehistory of the Eurasian starling clade, it is somewhat surprising that it is now the only extant African lineage in thisgroup. Pastor is another highly nomadic taxon that moves
widely throughout western and central Asia in response toephemeral foraging conditions, and that migrates to peninsular India for the winter (Feare and Craig, 1999).
4.1.5. Sturnus Linnaeus 1758
The basal split in the Eurasian starling clade separates
Sturnus vulgaris/unicolor from the remaining taxa. The
Sturnus lineage has the westernmost natural distributionof any Eurasian starling taxon, and Sturnus exhibits manyof the behavioral attributes (such as colonial breeding,dense roosting and foraging flocks, and nomadic/migratory
resource tracking) seen broadly among the species in theEurasian clade, suggesting that these traits are ancestral
to the entire group (Feare and Craig, 1999). The internodeseparating Sturnus from the remaining Eurasian starlings isnotably long and well supported (Fig. 1). Our sample ofSturnus vulgaris (from North American) was nearly genetically identical to our sample of S. unicolor (from Europe),
which is perhaps not surprising given that these species are
known to hybridize in an area of recent contact (summarized in McCarthy, 2006). Nonetheless, Sturnus vulgaris
has a broad geographic range and shows high geographic
variation in body size, mensural characters, feather iridescence, and plumage spotting (Feare and Craig, 1999);
although no detailed phylogeographic studies have yet
been conducted on Sturnus, this genus may contain substantial diversity.
An advantage of the generic classification proposed hereis that it recognizes genera with roughly similar magnitudes
of genetic divergence, and hence similar periods of phylogenetic independence. A disadvantage is that it retains fourmonotypic genera and four genera that contain only two
species. As noted above, a simpler taxonomy that also follows the monophyly criterion could lump our genera Acri-Author's personal copy

I.J. Lovette et al. / Molecular Phylogenetics and Evolution 47 (2008) 251–260

dotheres/Sternopastor, Temenuchus/Leucopsar/Sturnia, andGracupica/Agropsar, respectively.
Although highly provisional owing to the negligible
within-species sampling in our study, the low differentiation within several clusters of congeneric species suggeststhat these forms are recently diverged. Within Acridotheres,
there are two sub-groups of three species each with notably
low levels of within-group sequence divergence. The first ofthese comprises taxa (cristatellus/grandis/albocinctus) withparapatric distributions in continental Asia; information
on reproductive isolation in any areas of overlap among
these forms would be a useful contribution to their systematics. The second group includes one widespread continental form (the jungle myna A. fuscus) and two island taxa
(javanicus and cinereus) that have often been lumped into
fuscus (Table 1). A. fuscus and A. javanicus may now interbreed in areas of the Malay Peninsula where javanicus has
recently been introduced (Kang et al. 1994). A final
instance of extreme sequence similarity is represented by
Sturnus vulgaris and S. unicolor, the latter also having often
been treated previously as one of the many sub-species(Dickinson, 2003) of vulgaris; these two taxa have similarly
low differentiation at allozyme loci (de la Cruz-Cardiel
et al., 1997) and hybridize in a region of recent contact(summarized in McCarthy, 2006). In all three groups ofclosely allied taxa, our sampling of only 1–2 individuals/
species precludes detailed analyses of the genetic distinctiveness of these complexes, but our results suggest thateither the taxa within each of these groups are very recently
derived from one another, or that they have experienced
recent gene-flow. These results therefore provide marginal
support for re-lumping these forms into fewer taxonomic
species. Comprehensive analyses of phylogeographic patterns and geographic variation in behavior and morphology are required to test this possibility more rigorously.
4.2. Implications of nested gene and multilocus trees
The heterogeneous composition of our sequence dataset
is an example of a likely trend in phylogenetic studies ofbirds and other taxa. Three factors make it probable thatan increasing number of phylogenetic studies will be basedon short mitochondrial sequences from some taxa, analyzed in combination with sequences from more diverse setsof loci from other samples. First, for taxa from whichrobust DNA samples are available, it has become standard
to investigate phylogenetic relationships using multiple loci(often from both organellar and nuclear genomes), and weare beginning to be able to analyze these multilocus
sequence data using powerful coalescent methods (e.g.,
Edwards et al., 2007; Ané et al., 2007), which will further
encourage the generation of datasets targeting large numbers of independent loci. Second, it is also becoming routine (Wandeler et al., 2007) to obtain short sequences(usually mitochondrial) from old or degraded samples suchas avian museum skin specimens that were not originallycurated with DNA preservation in mind; in this study,

257

for example, we added a number of starling taxa based
solely on mtDNA sequences derived from toe-pads ofavian study skins that ranged in age from several decades
to more than a century (Table 2). The ability to use traditional museum specimens as a DNA source greatly
enhances our ability to generate comprehensive phylogenies for entire clades because many groups of organisms
are so broadly distributed that no individual worker could
collect field samples from all species, and many species are
likewise rare, endangered, extinct, or found only in politically or logistically inaccessible locations. The third factor
that will likely drive the generation of heterogeneous DNA
sequence datasets is the rapidly expanding generation of‘‘DNA barcodes,” typically short sequences of the mitochondrial COI gene, from most or all taxa in particulargroups or from defined geographic regions (e.g., Hebert
et al., 2004; Kerr et al., 2007). Although the information
content of these short sequences may often render DNA
barcodes sub-optimal for reconstructing phylogenetic relationships, they are already being broadly employed for thispurpose. Placing DNA barcode samples from a subset of
taxa within a topology based on a set of broader, more
phylogenetically informative markers has the potential togreatly expand the phylogenetic utility of barcode
sequences and databases.
As with all estimates of phylogenetic relationships basedon individual loci it is important to distinguish between
organismal histories and the gene trees derived from individual loci. In this study, the mitochondrial gene tree was
highly similar to the multilocus topology for the species
represented in both datasets, but this will not always be
the case. It is important to keep in mind that a numberof taxa and nodes in our multilocus reconstruction (thinbranches in the left-hand tree in Fig. 1) are placed there
solely via their mitochondrial gene tree. Distinguishing
between single gene tree topologies and multilocus topologies will become increasingly challenging as heterogeneous
datasets of this type expand to include large number oftaxa and loci.
4.3. Trait variation and evolution
These Eurasian starlings figured prominently in Beecher’s (1978) detailed comparative study of starling skull
morphology, an investigation initially motivated by hisobservation that the jaw musculature and skull shape of
the European starling (Sturnus vulgaris) are both highly
modified for prying. The European starling typically forages by first forcing its bill into a turf mat then prying open
the bill to create a window in the turf within which it scans
for invertebrate prey (Feare, 1984). The force required toopen the bill against pressure is opposite that faced by mostother birds, for which the greatest force is required when
closing the bill (as when cracking a seed or holding prey).
Beecher (1978) found that the adductor muscles used toopen the bill are correspondingly robust in the European
starling, and that the anterior region of the skull is further

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modified to facilitate forward vision along the axis of thebill, which is necessary for spotting prey in the hole heldopen by the bill. In extending this investigation to otherstarling genera, Beecher found that these modifications
were absent in most starling taxa, but documented a graded
series of specialization within the Eurasian starlings. Beecher placed the Eurasian starling species into three groups
based on their degree of prying-related modification, andhypothesized that each of these groups had originated independently from a common, forest-dwelling (and non-prying) ancestor. Beecher’s morphological groupings haveinfluenced all later classifications of this group (e.g., Wolters, 1982; Sibley and Monroe, 1990; Dickinson, 2003).
In mapping Beecher’s categorization of each species
onto our molecular tree (Fig. 2), we found that the skull
traits he documented are poorly conserved. The few taxa
with morphologies highly specialized for prying (Sturnus
vulgaris/unicolor and Poliopsar cineraceus) are not close
allies: Sturnus is the sister to the remaining Eurasian starlings, and P. cineraceus is embedded within a clade of taxa
that all show only moderate prying specialization. This
most extreme form of prying-related modification therefore
likely evolved twice within this group. Likewise, the two
taxa (Temenuchus pagodarum and Pastor roseus) that are

Skull Morphology
Low
Intermediate
ExtremeMovement
Sedendary
Local movements
Highly migratory
or nomadic

similar to non-Eurasian starlings in exhibiting low skull
specialization are not phylogenetically close, and each isallied to a group of taxa with moderate prying specialization (Fig. 2). These patterns suggest that these morphological traits have high evolutionary lability, and that theyshould be employed with caution as characters for phylogenetics and classification.
High lability in behavioral and life history traits has
been shown previously in African starlings (Rubensteinand Lovette, 2007), but the Eurasian starlings have highconservation of some behavioral traits. All 26 Eurasiantaxa are gregarious; most forage and roost in large flocks
during the non-breeding season, and many breed colonially(Feare and Craig, 1999). Nearly all taxa are habitat generalists, occurring in a variety of disturbed habitats; only
Leucopsar, and to a lesser degree Sturnornis and Agropsar
sturninus, are restricted to non-disturbed forest habitats.
More variation is present in movement behavior (Fig. 2):
just under half of the Eurasian starling species are sedentary, whereas the rest show a gradient of movement types
ranging from local migration, to periodic nomadicism, toobligate long-distance migration. There is low phylogenetic
signal in the distribution of migratory behavior, as obligate
migration appears to have arisen independently in four

Acridotheres cristatellus
Acridotheres grandis
Acridotheres albocinctus
Acridotheres fuscus
Acridotheres cinereus
Acridotheres javanicus
Acridotheres melanopterus
Acridotheres burmannicus
Acridotheres ginginianus
Acridotheres tristis
Poliopsar sericeus
Poliopsar cineraceus
Temenuchus malabaricus
Temenuchus erythropygius
Temenuchus pagodarum
Temenuchus sinensis
Leucopsar rothschildi
Sturnornis albofrontata
Gracupica nigricollis
Gracupica contra
Agropsar philippensis
Agropsar sturninus
Pastor roseus
Creatophora cinerea
Sturnus unicolor
Sturnus vulgaris

Fig. 2. Phylogenic distribution of variation in skull morphology and movement behavior among Eurasian starling species. Tree is a phylogram based on
the reconstructions shown in Fig. 1. Specialization in skull morphology has historically contributed importantly to the classification of these taxa, andspecies are coded into three skull morphology classes (extreme, moderate, and low specialization for prying) following Beecher (1978). Information on
species’ tendency to be sedentary, locally, or more broadly migratory/nomadic is derived from Feare and Craig (1999). Species not studied by Beecher(1978) or with unknown movement behavior were not given boxes.

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Eurasian starling lineages (Fig. 2). The lability of starling
movement behavior is strikingly illustrated by the rapid
evolution of migration in the European starling populationof eastern North America following its introduction in1890 (Kessel, 1953).
A combination of these traits seen broadly in the Eurasian clade—use of disturbed habitats, gregarious association, and behavioral lability—has likely contributed to thehuman-mediated spread of Sturnus vulgaris and Acridotheres tristis around the globe. Many other Eurasian clade
species are expanding their geographic ranges as anthropogenic modification increases the amount of suitably disturbed habitat. It is notable and somewhat paradoxicalthat the European starling (which may now have the most
expansive geographic range of any passerine bird) and its
ecologically successful relatives fall within a recent radiation that also contains several highly endangered singleisland endemics.
Acknowledgments
We thank Paul Sweet and Joel Cracraft of the American
Museum of Natural History, Carla Cicero of the Museumof Vertebrate Zoology at the University of California,
Berkeley, and Jeremiah Trimble and Scott Edwards ofthe Museum of Comparative Zoology at Harvard University for granting us permission to sample toe-pads fromstudy-skin specimens in their respective collections. We
also thank the institutions that provided tissue samples
from their collections: American Museum of Natural History; US National Museum of Natural History (Smithsonian); Museum of Natural Science, Louisiana State
University; Burke Museum, University of Washington;
Cornell University Museum of Vertebrates; Muséum National Histoire Naturelle, Parc de Clères. This research
was supported by the National Science Foundation(DEB-0515981).
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