A MOLECULAR PHYLOGENY of the DOVE GENERA <I>

Home , Columba (bird), Macropygia, Nesoenas, Patagioenas, Pink pigeon, Reinwardtoena

The Auk 118(4):874-887, 2001

A MOLECULAR PHYLOGENY OF THE DOVE GENERA STREPTOPELIA AND COLUMBA

KEVIN P. JOHNSON,L3,s SELVINO DE KORT,2 KAREN DINWOODEY,3 A. C. MATEMAN,4 CAREL TEN CATE,2 C. M. LESSELLS,4 AND DALE H. CLAYTON3 •IllinoisNatural History Survey,Champaign, Illinois 61820, USA; 2Instituteof Evolutionaryand Ecological Sciences, Leiden University, Leiden, The Netherlands; 3Departmentof Biology,University of Utah,Salt Lake City, Utah84112, USA;and 4NetherlandsInstitute of Ecology,Heteren, The Netherlands

ABSTRACT.--Evolutionaryhistory of the dove genus Streptopeliahas not been examined with rigorousphylogenetic methods. We presenta study of phylogeneticrelationships of Streptopeliabased on over3,600 base pairs of nuclearand mitochondrialgene sequences. To testfor monophylyof Streptopelia,we usedseveral other columbiform taxa, including Colum- ba(Old and New World),Macropygia, Reinwardtoena, and the enigmaticPink Pigeon(Nesoenas mayeri).On the basisof our analyses,Streptopelia (as currently defined) is not monophyletic; Nesoenasmayeri is the sisterspecies to S.picturata, resulting in paraphylyof Streptopelia.Three main cladesof Streptopeliaare identified: (1) S. chinensisplus S. senegalends,(2) S. picturata plus Nesoenasmayeri, and (3) all other speciesof Streptopelia.It is unclearwhether thoseclades form a monophyleticgroup to the exclusionof Old World Columba,but severalanalyses pro- ducethat result.Species of Old World Columbaare closelyrelated to Streptopelia,with species of New World Columbaclustering outside that group.Taxonomic changes suggested by our resultsinclude merging Nesoenaswith Streptopeliaand changingthe genericname for New World Columbaspecies to Patagioenas.Vocal similarities between S. picturata and N. mayeriare striking, given the generaldiversity of vocalizationsin other species.Received 20 September 2000, accepted27 March 2001.

SPECIESOF DOVES in the genusStreptopelia are lationshipswithin the genusare uncertain.No- important model systemsfor studiesof physi- wak (1975)examined morphological character- ology (Walker et al. 1983, Janik and Buntin istics within Streptopeliaand produced a 1985, Cheng 1986,Ramos and Silver 1992, ten classificationthat grouped speciesinto several Cateet al. 1993,Lea et al. 1995,Georgiou et al. subgenera.However, he did not producean ex- 1995) and behavior (Lade and Thorpe 1964, plicit phylogenetictree. Goodwin (1983) de- Zenoneet al. 1979, Cheng et al. 1981, Cheng picts a tree of "presumed relationships" 1992, Slabbekoorn and ten Cate 1998, Slabbe- among speciesof Streptopelia,and that tree dif- koornet al. 1999).The 16 speciesof Streptopelia fers from Nowak's classification, but Goodwin's historically had an African and Eurasian dis- tree is not based on a rigorous phylogenetic tribution, but some specieshave been intro- analysis.Johnson and Clayton (2000a)showed duced to the New World and Australia. Several that speciesof Old World Columbaform the sis- speciesof Streptopeliaseem highly adaptableto ter group to Streptopelia,with speciesof New human-altered environments and have ex- World Columbabeing more distantly relatedto panded their ranges considerably(e.g.S. de- both groups. However, the authors included caocto;Bijlsma 1988, Hengeveldand van den only single representativesof Streptopeliaand Bosch1991, Hengeveld 1993, Kasparek 1996). Old World Columbain their study.The goal of Other specieshave remarkably localized distri- the presentstudy is to assessmonophyly of the butions(e.g.S. hypopyrrha).An understanding genus Streptopeliaand identify relationships of historical relationshipswould provide an within the genus.We alsouse this study to test importantcontext for work on physiology,be- Nowak's (1975) classification and Goodwin's havior,and biogeographyof this genus. (1983)proposed relationships of specieswithin Eventhough species of Streptopeliahave been the genus.We include severalrepresentatives well studiedin manyrespects, phylogenetic re- of New and Old World Columba, as well as the endangered,enigmatic Pink Pigeon (Nesoenas E-mail: [email protected] mayeri)of Mauritius. We alsoinclude represen-

874 October2001] Phylogeny0f Streptopelia 875 tatives of Macropygiaand Reinwardtoena,which of individual variation within each species,we se- togetherwith Columbaand Streptopeliaform a quenced (when available) multiple individuals for distinct clade within Columbiformes (Johnson the COI gene (see Table 3 for list of those multiple and Clayton 2000a). The entire phylogenetic individuals and GenBank accessionnumbers). Mi- tochondrial genes tend to evolve at similar rates at analysisis rooted on Geotrygon,Leptotila, and low levelsof sequencedivergence in birds (Johnson Zenaida,which together consitute the sister and Sorenson1998, Johnsonand Lanyon 1999) and group to the Columba/Streptopelia/Macropygiamore specificallyfor cyt b, ND2, and COI in Col- clade (Johnsonand Clayton 2000a). umbiformes (Johnson and Clayton 2000b). Mito- For our current study, we use sequencesof chondrial divergencesare also highly correlated both mitochondrial and nuclear genesto con- with divergencesin FIB7 in Columbiformes(Johnson structa phylogenyfor Streptopelia,Columba, and and Clayton 2000a, b). Thus, within-speciesvaria- relatedtaxa. We comparerelative usefulness of tion in COI sequencesis likely to be representativeof nuclearand mitochondrialgenes for phyloge- other mitochondrialand nuclear genes.In general, netic resolutionat that level.Using the phylog- individualsof the samespecies differed only slightly, if at all, in thoseCOI sequences(see below), so single eny we recommendsome changes in taxonomic exemplar individuals are reasonablefor the multi- classification,and comparephylogeny to vocal gene data set. similarities and diversity in Streptopelia. Where possible,we used tissueor feathersamples to avoid risk of nuclear copies of mitochondrial METHODS genes (Sorensonand Quinn 1998). In caseswhere blood sampleswere used,we verified sequencesfor Sequencing.--Weobtained samplesof muscle tis- COI using multiple individuals. We also checked sues,feathers, or blood from representativesof 14 of chromatogramsfor signsof doublepeaks, as well as the 16 describedspecies of Streptopelia(Table 1). We checkingfor indels and stop codons.By sequencing also sampled members of New and Old World Co- several mitochondrial genes, we were also able to lumba,Macropygia, Reinwardtoer•a, Nesoenas, Geotry- te•t for any incongruencein phylogeniesresulting gon,Leptotila, and Zenaida.We extractedDNA from from thosegene regions, which would likely occurif those samplesusing a Qiagen Tissue Kit (Valencia, some of the sequencesrepresented nuclear copies. California) with the manufacturer'sprotocols. For We found no evidence of nuclear copies in our feather samples(-2 mm of the tip of the shaft) we sequences. alsoadded 30 btLof 10% DTT to the digestionbuffer. Phylogeneticanalysis.--Relative rates of substitu- Using PCR we amplifiedportions of three mitochon- tion can be examined by plotting pairwise sequence drial genes:cytochrome-b (cyt b), cytochromeoxi- divergences for various substitution types and dase I (COI), and NADH dehydrogenasesubunit 2 genes.For the nuclear gene(FIB7), we estimatedthe (ND2). We also amplified the nuclear gene [3-fibrin- "native" transition:transversionratio by plotting ogen intron 7 (FIB7). Table 2 lists primers used for pairwise transition differences against pairwise those reactions. Protocols for reactions follow John- transversiondifferences. Slope of the linear portion son and Clayton (2000a). of that curve estimates the transition:transversion ra- Direct sequencingof PCR products and determi- tio (Sturmbauer and Meyer 1992). We do not fit re- nation of sequenceswas performed as describedby gressionsthrough those points because they are non- Johnsonand Clayton (2000a).We aligned sequences independent, rather we use slopes as a rough acrossspecies using Sequencher(GeneCodes, Mad- indicator of relative rates. For mitochondrialgenes, ison, Wisconsin).The three mitochondrialgenes to- we plotted transition differenceat third positions taled 2,520 bp and the nuclear intron included 1,150 against transversionsat third positionsto estimate aligned base pairs (GenBankaccession numbers in that ratio. To estimate relative rates of mitochondrial Table 1). In the caseof mitochondrialprotein coding versusnuclear substitution, we plotted pairwise di- genes,alignment was straightforward.We noted sev- vergencesfor mitochondrialgenes against those for eral insertion-deletion (indel) events in the nuclear FIB7. We also plotted pairwise divergencesfor in- intron, but for those taxa, divergenceswere so low dividual mitochondrialgenes against each other and that manual alignment was straightforward.Gaps againstFIB7 to determinewhich genesare more sub- were treated as missingdata in phylogeneticanaly- ject to multiple substitution. ses. For Streptopeliatranquebarica and S. bitorquata, PAUP* (Swofford2000) was used for all phyloge- only partial FIB7 sequenceswere availablebecause of netic analyses.We used the partition homogeneity amplificationfailures. Thus, we repeatedanalyses test (Farris et al. 1994, 1995; Swofford 2000) to ex- involving the nuclearintron both with and without aminewhether the generegions could be considered thosespecies. samples of the same underlying phylogeny, or For each species,we included single representa- whether there was evidencefor different phyloge- tives for all sequences.However, to assessthe level netic signal between gene regions.Even though that 876 JOHNSONETAL. [Auk,Vol. 11• October2001] PhylogenyofStreptopelia 877

TABLE2. Primers used in study.

Gene Primer name Source cyt b L14841 Kocher et al. (1989) H4a Harshman (1996) H15299 Kocher et al. (1989) L15517 Johnsonand Sorenson (1998) ND2 L5215 Hackett (1996) H6313 Johnson and Sorenson (1998) L5758 Johnsonand Sorenson (1998) COI L6625 Hafner et al. (1994) H7005 Hafner et al. (1994) FIB7 FIB-BI7L Prychitko and Moore (1997) FIB-BI7U Prychitko and Moore (1997) FIB-DOVEF Johnsonand Clayton (2000a) FIB-DOVER Johnsonand Clayton (2000a) test indicated no significantincongruence between dall 1997). Using the estimatedmodel parameters, any of the gene regions (see below), we also con- we conducted 50 heuristic maximum-likelihood ducted severalanalyses with mitochondrialand nu- searcheswith nearest neighbor interchange (NNI) clear genesseparately. We did this becauseof the branch swapping.We used bootstrapping(100 rep- slow rate and lack of informative sites in the nuclear licateswith NNI branchswapping) to evaluaterela- geneand becausewe wantedto identifywhat nodes tive support for nodes in maximum-likelihood were common to trees derived from independently topologies. sorting gene regions. We conductedparsimony analyses with a range of RESULTS transversionweighting schemes from 1:1(unordered parsimony)to 20:1. For eachweighting scheme,we Sequence variation.--Mitochondrial genes used 50 random addition replicateswith heuristic were similar in the fraction of sites that were searches.For eachanalysis, we alsoperformed 1,000 variableand potentiallyphylogenetically infor- bootstrapreplicates (Felsenstein 1985) to assessrel- mative, but ND2 showed the most variation, ative supportfor variousbranches in the phylogeny. We used one of the combined unordered parsi- followed by cyt b, then COI (Table 4). FIB7 showed a much smaller fraction of variable and mony trees to estimate parametersfor maximum- likelihood searches. We used likelihood ratio tests to phylogeneticallyinformative sites (Table 4). estimate the best fit model that could not be rejected Within Streptopelia,mitochondrial sequence di- in favor of a simplermodel (Huelsenbeckand Cran- vergencesranged from 1.9 to 12.4%.For those

TABLE3. Samplessequenced for COI only.

GenBank Species Locality Samplenumber Tissuetype number Streptopeliadecaocta Netherlands SDK I Feather AF353495 Streptopeliadecaocto Netherlands SDK2 Feather AF353496 Streptopeliaroseogrisea Cameroon SDK2 Blood AF353497 Streptopeliaroseogrisea Cameroon SDK3 Feather AF353498 Streptopeliadecipiens Cameroon SDK2 Feather AF353499 Streptopeliasemitorquata Cameroon SDK1 Feather AF353500 Streptopeliacapicola SouthAfrica LSUMNSB34205 • Muscle AF353501 Streptopeliavinacea Cameroon SDK 1 Feather AF353502 Streptopeliahypopyrrha Cameroon SDK2 Feather AF353503 Streptopeliaturtur Cameroon SDK 1 Feather AF353504 Streptopeliaturtur Cameroon SDK 2 Feather AF353505 Streptopeliaorientalis Captive SDK 2 Feather AF353506 Streptopeliatarnquebarica Captive SDK3 Feather AF353507 Streptopeliapicturata Captive SDK 1 Feather AF353508 Streptopeliachinensis Singapore AMNH PRS-678a Fuscle AF353509 Streptopeliasenegalensis Cameroon SDK 1 Feather AF353510 Indicatessamples that havea correspondingmuseum skin voucher. 878 JOHNSONETAL. [Auk,Vol. 118

TABLE4. Variableand phylogeneticallyinformative 0.25 sitesfor eachgene region.

Number Percent Percent Gene of sites variable information cyt b 1070 39.0% 33.1% ND2 1067 48.7% 39.8% COI 383 34.2% 30.0% FIB7 1150 19.6% 7.9%

same comparisons,FIB7 showed much less divergence(0.0 to 1.4%). Plots of third position transitionsagainst third position transversions 0 i i i for mitochondrialgenes (Fig. 1) indicatedcon- 0 0.025 0.05 0.075 0.1 siderablemultiple substitutionof third position transitions with an estimated transition: mitochondrialthird position transversions transversion ratio of approximately 10:1. FIG.1. Plot of pairwisepercentage divergence for Divergencesof mitochondrialgenes were sim- transitionsagainst transversions for third positions ilar, but comparisonsof thosedivergences in- of all mitochondrialgenes combined. dicatedthat ND2 was lesssubject to multiple substitutionthan was cyt b or COI, because plots of ND2 divergenceagainst FIB7 or the drial genesindicate that those genescould be other mitochondrialgenes indicated continu- consideredsamples of thesame underlying phy- ing accumulationof substitutionsat high di- logeny(P = 0.44).Similarly, trees derived from vergences.The estimated transition:transver- FIB7 alonecompared to treesfor mitochondrial sion ratio for FIB7 was approximately 1.5:1 genescombined were not significantlyincon- (Fig. 3), with no evidenceof multiple substi- gruent (P = 1.0). tution of transitions compared to transver- Unordered parsimony analysisof all three sions. Plots of divergencesfor the mitochon- mitochondrialgenes combined produced three drial genesagainst FIB7 divergenceindicated a equally parsimonioustrees (not shown,but see 5 to 10x higher rate of substitution in the mi- Fig. 5 for reference).Those trees indicatedthat tochondrialgenes (Fig. 4). Streptopeliais not monophyletic.Nesoenas mayeri Several indels occurred in FIB7. One of those is the sistertaxon to S.picturata with high boot- is a 166 bp deletionshared by Stre?to?elia,Ne- strap support.In addition,two groupsof Strep- soenas, and Old World Columba. Other colum- topeliaare sister to Old World Columba,but that biform taxa do not show that deletion. Other in- result has only weak bootstrapsupport (49%). delsin FIB7 rangedfrom i to 125bp, but only One large groupof Streptopeliais monophyletic two others were potentially phylogenetically (clade A) with strong support (100%). informative(one uniting Stre?to?elia,Nesoenas, Transversionweighting of 2:1 to 5:1 pro- Old World Columba,and Macro?ygiathe other duced identicaltree topologies(one tree) that uniting cladeA of Stre?to?elia[Fig. 5]). were completelyresolved (not shown).Analy- Within species,little sequencevariation was sis of mitochondrial genes with those low evident.For the COI gene,within-species vari- transversionweights also produced a paraphy- ationranged from 0.0% (for 6 of 13 specieswith letic Streptopelia,both with respectto Nesoenas multiple samples)to 1.0% (for comparisonsof and with respectto Old World Columba.How- Strepto?eliasemitorquata between South Africa ever,transversion weighting of 10:1and higher and Cameroon).The low within-speciesvaria- produceda singletree showingmonophyly of tionfor mitochondrialgenes suggests that single Streptopeliawith respectto Old World Columba. speciesexemplars are suitablefor species-level Nesoenaswas still sister to S. picturata,again phylogeneticanalysis, given that between-spe- with high bootstrapsupport. ciesdivergences all exceeded2.5%. Unorderedparsimony analysis of the nuclear Phylogeny.Partition homogeneity tests com- intron alone produced 32,020 trees (not paring congruenceamong the three mitochon- shown). The high number of trees was not sim- October2001] PhylogenyofStreptopelia 879

0.2 0.15

a o© O 0.15

0.1

.05

0.05 g

o 0 0.05 0.15 o

cytb percentdivergence FIB7 percentdivergence

0.2 0.2

0.15 0.15

o.1 E!o 0.1

o.o5 0.05

0 0 i [ i 0 0.;5 011 d5 012 0 0.02 0.04 0.06 0.08

COI percentdivergence FIB7 percentdivergence

0.15 0.2

0.05 o r,.) 0.05 OL•/• • , • 0 0.;5 011 0.[5 0.2 0 0.02 0.04 0.06 0.08 COI percentdivergence FIB7 percentdivergence FIG.2. Plot of pairwisepercentage divergence for (A) ND2 versuscyt b, (B) ND2 versusCOI, (C) cyt b versusCOI, (D) cyt b versusFIB7, (E) ND2 versusFIB7, (F) COI versusFIB7. Lines for the mitochondrialgene plots(A-C) indicateexpectation if ratesare equal. 880 JOHNSONET AL. [Auk,Vol. 118

0.2 0.05[ 0'041 81:1[] .•0.15 ß• ß ,• o.1

.• 0.05

i i 0I• i i I 0 0.005 0.01 0.015 0.02 0.025 0.03 0 0.02 0.04 0.06 0.08

FIB7 transversions FIB7 percentdivergence

FIG.3. Plot of pairwisepercentage divergence for FIG. 4. Plot of overall percentagedivergence for transitionsagainst transversions for all positionsof all mitochondrialgenes combined against percent- nuclear FIB7. age divergencefor nuclearFIB7. ply an artifact of incomplete sequencesfor els. Maximum-likelihood searches on mito- Streptopeliabitorquata and S. tranquebarica.Ex- chondrial genes alone produced a tree very clusionof thosesequences from phylogenetic similar to parsimonytrees, and that tree indi- analysesproduced 4,236 trees,the consensusof cated monophylyfor Streptopeliawith respect which was identical to the consensus of trees to Old World Columba.Likelihood analysis of with those taxa included. FIB7 trees did not re- the intron alone identifiesmonophyly for the solve relationshipsamong major Streptopelia three main cladesof Streptopeliaand Old World lineages plus Old World Columba.However, Columba,but does not resolve relationships New World Columbais placed outsideStrepto- among those groups. There is also little reso- peliaand Old World Columba(bootstrap sup- lution within the large Streptopeliaclade A. port 100%).Monophyly of both cladesB and C Maximum-likelihoodtrees for combinedgene of Streptopelia(see Fig. 5 for reference)was regions (Fig. 6) are well resolvedand indicate stronglysupported in parsimonyanalyses of monophyly for Streptopelia(inclusive of Nesoen- FIB7 alone, as was monophyly of Old World as) with moderate support (bootstrap 64%). Columba. The three main cladesof Streptopeliaidentified Unorderedparsimony analysis of combined by combinedparsimony analysis are also re- gene regions (mitochondrialand nuclear)pro- coveredin that tree with strongsupport (100% duced two trees (Fig. 5). Strict consensusof in eachcase), as is paraphylyof Columba(100% thosetrees, like unorderedparsimony analysis bootstrapsupport for sister relationshipbe- of mitochondrialgenes alone, does not recover tween Old World Columbaand Streptopelia).In a monophyleticStreptopelia with respectto Old fact, supportfor a sisterrelationship between World Columba.However, in mostrespects that New World Columba and Old World Columba + tree is very similar to treesproduced by sepa- Streptopeliais relativelypoor (51%).Most other rate analysis.Increasing transversion weight- branchesare identical to thoserecovered by un- ing of combinedgene regions (in excessof 10: ordered parsimonyanalysis (Fig. 5). 1) produced trees with a monophyletic Streptopeliawith respectto Old World Columba DISCUSSION (not shown, see Fig. 6 for reference). Likelihood ratio tests indicated that models Phylogeny.--Aphylogeny based on 2,530bp incorporating unequal base frequencies,six of mitochondrialand 1,150bp of nuclearDNA substitutiontypes, and rateheterogeneity were sequencesis generally well resolved and well statisticallybetter justified than simplermod- supported for the dove genus Streptopeliaand October2001] Phylogenyof Streptopelia 881

• Streptopeliaroseogrisea 11•_••StreptopeliaStreptopeliadecipiens decaocto 59 Streptopeliacapicola 100t Streptopeliavinacea Streptopeliasemitorquata Clade A

lOO 10o Streptopeliaturtur l• StreptopeliaStreptopeliaorientalis hypopyrrha Streptopeliabitorquata Streptopeliatranquebarica • 100 F Columbalivia.• 69] C---Columba rupestris I 74 ! Columbaguinea / OldWorld I - ColumbaColumbaarquatrixpalumbus I Columba Columbapulchricollis •! Nesoenasmayeri 3 I 100100[ I StreptopeliaStreptopeliaStreptopeliapicturatachinensissenegalensis 3 Clade BC 86 Columbaleucocephala w

66 100 Columbaplumbea "Columba" 99 • ••oo Columbaspeciosa New World lOO • ] Columbasubvinacea (Patagioenas) ' Columbaoenops Columbafasciata Macropygiamackinlayi lOO 1© I Macropygia tenuirostris Reinwardtoena browni Leptotilarufaxilla lOO[ Leptotilaverreauxi

100[ ZenaidaZenaidaasiatica macroura Geotrygonmontana

FIG. 5. Strict consensusof two trees (length = 4470, RC = 0.200) from unorderedparsimony analysis of combinedmitochondrial (cyt b, ND2, COOand nuclear(FIB7) genes.Numbers above branches indicate bootstrap supportfrom 1,000full heuristicreplicates. Unlabelled nodes received <50% bootstrapsupport. Branch lengthsare proportionalto reconstructedchanges using parsimony.Indicated groups are cladesreferred to in text.

outgroup taxa. However, even given the rela- better resolutionfor the phylogenyof Streptope- tively large amount of DNA sequencedata, lia. Mitochondrialgenes have around a 5 to 10x some aspectsof the tree are still weakly sup- higher substitutionrate (Fig. 4), and thus vari- ported. Primary among those uncertaintiesis ation can accumulate in mitochondrial DNA be- whether or not Streptopeliais monophyletic tweenclosely related species. For several closely with respectto Old World Columba. relatedspecies of Streptopelia,sequences of FIB7 Despite the lower homoplasypresent in the were identical,preventing resolution of species FIB7 sequences,mitochondrial genes provided level relationships.Importantly for this study, 882 JOHNSONET AL. [Auk, Vol. 118

• Streptopeliadecaocto i Streptopeliaroseogrisea 100••• Streptopeliadecipiens Streptopeliasemitorquata lO0 Streptopeliacapicola Streptopeliavinacea Clade A Streptopeliahypopyrrha Streptopeliaturtur Streptopeliaorientalis 64 Streptopeliabitorquata

Streptopeliatranquebarica i -- Nesoenasmayeri i Clade B 100 {• Streptopeliapicturatai i 100 Clade C 100 I StreptopeliaStreptopeliachinensissenegalensis i Columba livia i

10• 100LColumba Columba guinea rupestris Old World Columba lOO ß Columbaarquatrix 8• [ ColumbaColumbapulchricollis palumbus Columbaplumbea

New World

lOO 100 Columbaoenops "Columba" lOO 1•00 96[ Columbasubvinacealeucocephala (Patagioenas) Columbaspeciosa Columbafasciata 100[ MacropygiaMacropygiatenuirostrismackinlayi lOOt Reinwardtoena browni 100 [ Leptotilarufaxilla 100 Zenaida asiatica Zenaida macroura 5• [ LeptotilaGeotrygonverreauximontana -- 0.01 substitutions/site

FIG. 6. Tree resulting from maximum likelihood searchesusing combinedmitochondrial (cyt b, ND2, COI) and nuclear(FIB7) genes(ln likelihood = -25,202; modelparameters: gamma shape parameter = 0.209 with eight rate categories,transition matrix: A-C = 1.41, A-G = 17.96, A-T = 1.22, C-G = 0.54, C-T = 16.77, G-T = 1.0). Numbers abovebranches indicate bootstrap support from 100 heuristicsearch replicates. Un- labelled nodesreceived <50% bootstrapsupport. Branchlengths are proportionalto likelihood estimated branch lengths (scaleindicated). Indicated groups are cladesreferred to in text. trees based on FIB7 identified the same major groups involve only two species:S. chinensis+ cladesthat were identifiedby analysesof mito- S. senegalensis(clade C) and S. picturata+ Ne- chondrial genes.In addition, there was no sig- soenas(Streptopelia) mayeri (clade B). The other nificant conflict between nuclear and mitochon- clade (A) includesall other speciesof Streptope- drial data sets,suggesting that treesrecovered lia. Monophylyof eachof thoseclades is gener- in our analysesestimate the species'phylogeny ally well supported,but relationshipsamong ratherthan simply genetrees. those groups and Old World Columbais less All analysesidentify threemajor cladeswith- clear.Analyses that takeinto accountlarge tran- in Streptopelia(Figs. 5 and 6). Two of those sition:transversionbiases (transversion-weight- October2001] Phylogenyof Streptopelia 883 ed parsimonyand maximum likelihood)tend to ranges from 11.1 to 11.9%, whereas the maxi- recovermonophyly for Streptopelia(inclusive of mum mitochondrial divergenceswithin clade Nesoenas). A of Streptopeliarange to 8.4%. Phylogeneticrelationships within Streptope- Streptopeliavinacea and S. capicolaare a pair lia do not correspondin a strongway to either of allopatric sister taxa of particular interest. Nowak's (1975) classification or Goodwin's Those taxa are distributed in sub-Saharan (1983) hypothesized evolutionary relation- northern Africa and in southeastern Africa, re- ships.However, similaritieswith both are evi- spectively. Another pair of dove taxa with a dent. Nowak (1975) unites S. decipiens,capicola, similar distribution is Turtur abyssinicusand T. vinacea,tranquebarica, semitorquata, roseogrisea, chalcospilos.Mitochondrial sequence diver- and decaoctointo a subgenus.We find support gencebetween S. vinaceaand S. capicolais 2.5% for a similar clade (excluding tranquebarica). and that between T. abyssinicusand T. chalcos- Goodwin (1983) outlines a similar group, but pilosis 1.7% (K. P. Johnsonunpubl. data). Sim- placestranquebarica outside of it and puts bitor- ilarity in geographicdistributions and genetic quataclose to decaocto.In our phylogeneticanal- divergencessuggests some common biogeo- yses, S. tranquebaricaand S. bitorquataconsis- graphic factor underlying speciationbetween tently fall at the baseof a cladecontaining those thosespecies of Streptopeliaand Turtur,perhaps two speciesplus S. orientalis,turtur, and hypo- -1 Mya. pyrrha. Like many speciesof Streptopelia,tran- Taxonomicimplications.--The Pink Pigeon, in quebaricaand bitorquatapossess a "ring-neck," the monotypic genus NesoenasSalvadori 1893, unlike orientalis,turtur, and hypopyrrhathat is sister to Streptopeliapicturata. That result is have a dark patch on the side of the neck. Our evidenced by two independent gene regions results suggestthat the ring-neck (a dark bar and receivesstrong support in all analyses.The running aroundthe backof the neck)is the an- sister relationship between Nesoenasmv•yeri cestralcondition in group A of Streptopelia,hav- (Mauritius) and S.picturata (Madagascar) is not ing been lost in the ancestorof S. orientalis,tur- surprising, given the geographicproximity of tur, and hypopyrrha.Species of Streptopelia thosetwo islands.In light of thoseresults, we outside group A tend to have spotting on the recommendtransferring N. mayerito the genus neck. The closerelationship of S. chinensisand StreptopeliaBonaparte 1855. S. senegalensiswas recognized by Goodwin Regarding the generic status of Streptopelia, (1983) and to someextent by Nowak (1975). our results do not conclusivelydemonstrate A novel finding of our study is the sisterre- monophyly of Streptopelia.However, in analy- lationship between the Pink Pigeon (Nesoenas ses that take into accountrate heterogeneity mayeri),which is endemicto Mauritius, and the and transitionbiases, Streptopelia (as redefined MadagascanTurtle Dove (Streptopeliapictura- to includeNesoenas mayeri) is monophyletic.On ta), endemicto Madagascar.In most classifica- the basisof thoseresults, we suggestretaining tions, Nesoenasis united with speciesof Old the name Streptopeliafor all speciescurrently in World Columbawhereas the MadagascanTurtle the genus. An alternative would be to recog- Dove is consideredto be an aberrant Strepto- nize the three main clades (A-C) as separate pelia.On the basis of the closerelationship of genera, but we feel a more conservativeap- thosetwo species,we suggestthat the common proach,minimizing name changes,is prudent. ancestorof the Pink Pigeon and the Madagas- All of our analysesindicated that Old World can Turtle Dove colonized Mauritius from Columbaspecies are sister to, or imbedded Madagascarand subsequentlyspeciated. Per- within, Streptopelia.That phylogeneticposition centagedivergence for mitochondrialgenes be- results in paraphyly for Columba.Our current tween the Pink Pigeon and the Madagascan study confirmspreliminary results of Johnson Turtle Dove is only 3.0%. On the basis of mi- and Clayton (2000a), which also indicated that tochondrial molecular clock calibrations for relationship. BecauseColumba for Old World birds (Klicka and Zink 1997), that representsa specieshas priority, we recommend transfer- colonizationtime of -1.5 Mya, much younger ring all speciesof New World Columbainto the than the age of Mauritius (Proag 1995). Mito- genus Patagioenas(as partially suggestedby chondrialdivergence between the Madagascan Johnston1962). Johnston (1962) suggestedthat Turtle Dove and other speciesof Streptopelia the Band-tailed Pigeon (Columbafasciata) is a 884 JOHNSONET AL. [Auk, Vol. 118 close relative of Old World Columba; however, Streptopeliapicturata our results strongly supported monophyly of New World "Columba,"inclusive of fasciata. Further analysis is needed to determine if all New and Old World "Columba"species cluster by biogeographicdistribution, but we suspect Streptopeliamayeri that will be the case. Vocal evolution.--Streptopeliavocalizations havebeen studied extensively (Lade and Thorpe 1964, Gaunt et al. 1982, Cheng 1992, Ballintijn and ten Cate 1997, Slabbekoorn and ten Cate 1998, Slabbekoornet al. 1999),making that ge- Streptopeliacapicola nus an important group for comparativestudies 1.2 of vocalizations. Slabbekoornet al. (1999) compared acousticsimilarity of Streptopeliaperch- 0.8 cooswith Goodwin'sideas on taxonomy(1983). 0.4 They concludedthat, at the specieslevel, there is Streptopeliavinacea little congruencebetween similarity in perch- • 2i 3i 4i (s) coos and phylogeneticrelatedness, suggesting FIG. 7. Sonogramsof selectedStreptopelia perch- that environmentalor socialfactors promote the coos.Note the similarity between S. picturataand S. relativelyrapid diversificationof perch-coovo- mayeri.Sonograms of S. capicolaand S. vinacearepre- calizations.Although a formal reanalysisis be- senttwo sisterspecies of smallergenetic divergence, yond the scopeof this paper, an informal com- but greaterdivergence in perch-coovocalizations. parison of acousticsimilarity in relation to our phylogenydoes not contradictSlabbekoorn et al.'s (1999) conclusions. Streptopeliaare quite homogeneousin morpho- The closephylogenetic relationship between logicalcharacteristics. In fact, oftenthe mostre- Streptopeliapicturata and S. "Nesoenas"mayeri is liableway to discriminatespecies in the field is somewhatsurprising on the basisof morpho- by their distinctive perch-coos. Generally logical characters,but is supported by vocal speaking,vocalizations in Streptopeliaseem to characteristics.First, the relationshipin general be evolutionarily labile comparedto morpho- between Streptopeliaand "Nesoenas"is sup- logical characteristics. ported by similarities in the ExcitementCry Two hypothesesexplain why Streptopeliapic- (Baptista 1997). More dramatically,the perch- turataand S. mayerishow the greatestdegree of coosof S.picturata and S.mayeri are very similar vocal similarity within Streptopelia(Slabbe- (Fig. 7), unlike comparisonsof many other sis- koorn et al. 1999). First, selectionmay favor di- ter pairs of Streptopelia.Two other sisterspecies vergence between vocalizations of sympatric pairs also show some similarity in vocaliza- species,because of a need to use a signalthat tions as compared to other Streptopeliaspecies: can be distinguishedfrom that of closelyrelat- vinacea-capicolaand turtur-hypopyrrha. Al- ed species (Nelson and Marler 1990). Unlike though thosespecies pairs group togetherin a other Streptopeliaspecies, S. picturataand S. similarity analysis, they can be separated un- mayeriare allopatric with all other congeners, ambiguouslyon the basis of acousticparame- perhapsreducing need for divergencein signal ters (Slabbekoornet al. 1999). Thus perch-coos structure. Second, environment and habitat of- of other sister speciespairs show structural ten play important roles in evolution of avian acousticdifferences (e.g. Fig. 7), which is not signals(Morton 1975,Wiley and Richards1978, the casebetween S.mayeri and picturata.The lat- Marchetti 1993, Endler and Th•ry 1996, John- ter instead show considerabledivergence in son 2000, Johnsonand Lanyon 2000). Both S. morphologicalfeatures, including a lack of any picturataand S. mayeripredominantly inhabit neck pattern and a reddish bill in S. mayeri. densely forested habitats through which their Those features,among others,are the reasons low-pitched vocalizations should propagate that S. mayerihas often been placed in a mono- further than high-pitched vocalizations(Mor- typic genus.In contrast,most other speciesof ton 1975, Wiley and Richards 1978, Ryan and October2001] PhylogenyofStreptopelia 885

Brenowitz 1985, McCracken and Sheldon 1997). mous reviewersprovided helpful commentson this Further evidencefor potential stabilizing selec- manuscript. We thank the DNA SequencingFacility tion on the perch-coosof those two species at the University of Utah, supportedin part by Na- comesfrom examinationof the relationshipbe- tional Cancer Institute grant #5p30CA42014.This work was supportedby NSF-CAREERaward DEB- tween body size and soundfrequency. In New 9703003 to D.H.C. World pigeons,a negativecorrelation exists between body weight and perch-coo frequency LITERATURE CITED (Tubaro and Mahler 1998).That relationshipis not apparent in the perch-coosof $. picturata BALLiNTIJN, M. R., AND C. TEN CATE. 1997. Sex dif- and S. mayeri,because S. mayeriis nearly twice ferencesin vocalizations and syrinx of the Col- as heavy as S. picturata,yet sound frequencies lared Dove (Streptopeliadecaocto). Auk 114:22-39. of the perch-coosare very similar (Fig. 7). One BAPTISTA,L. 1997.Family Columbidae(Pigeons and or both of these factors may have caused vo- Doves). Pages60-243 in Handbook of the Birds calizations of those two speciesto remain rel- of the World, vol. 4. Sandgrouseto Cuckoos(J. del Hoyo, A. Elliott, and J. Sargatal,Eds.). Lynx atively unchangedsince speciation. Edicions,Barcelona, Spain. One other unexpectedphylogenetic relation- BIJLSMA,R. G. 1988. Populationgrowth in the Col- ship is also supportedby careful examination lared Dove Streptopeliadecaocto in the Nether- of vocalizations.Exclusion of Streptopeliabitor- lands. Limosa 61:41-42. quataand S. tranquebarica,despite similarity in CHENG,M. E 1986.Female cooing promotes oviarian plumage characteristics,from the group of typ- developmentin Ring Doves(Streptopelia risoria). ical ring-neckeddoves (decaocto through vinacea Physiologyand Behavior37:371-374. clade) is somewhatsurprising. The Excitement CHENG, M. F. 1992. For whom does the female dove or Flight Call is a conspicuouscharacteristic for coo? A case for the role of vocal self-stimulation. Animal Behaviour 43:1035-1044. the true ring-neckedspecies (as defined above). CHENG, M. F., M. PORTER, AND O. BALL. 1981. Do That display, in the same context, is absent in Ring Doves copulate more than necessaryfor S. bitorquataand S. tranquebarica.Absence of fertilization? Physiology and Behavior 27:659- that displayis sharedwith S.hypoppyrha, turtur, 662. and orientalis,the close relatives of bitorquata ENDLER,J. A., AND M. THI2RY.1996. Interacting ef- and tranquebarica.Likewise the bow-coo of fectsof lek placement,display behavior, ambient those two speciesis very similar to that of S. light, and color patterns in three Neotropical hypoppyrha,turtur, and orientalis(S. de Kort un- forest-dwellingbirds. American Naturalist 148: publ. data). In sum, despite the overall diver- 421-452. sity and rapid evolution of vocalizations in FARRIS, J. S., M. KALLERSJO,A. G. KLUGE, AND C. BULT.1994. Testingsignificance of congruence. Streptopelia,some behaviors reflect phylogenet- Cladistics 10:315-320. ic relationships. FARRiS, J. S., M. KALLERSJO,A. G. KLUGE, AND C. BULT.1995. Constructinga significancetest for ACKNOWLEDGMENTS incongruence.Systematic Biology 44:570-572. FELSENSTEIN,J. 1985. Confidencelimits on phyloge- We are grateful to the following individuals and nies: An approach using the bootstrap. Evolu- institutions (and curators thereof) who were instru- tion 39:783-791. mental in providing samples:American Museum of GAUNT, A. S., S. L. L. GAUNT, AND R. M. CASEY.1982. Natural History, Field Museum of Natural History, Syringeal mechanisms reassessed: Evidence University of Kansas Museum of Natural History, from Streptopelia.Auk 99:474-494. Louisiana State University Museum of Natural Sci- GEORGIOU,G. C., P. J. SHARP, AND R. W. LEA. 1995. ence,U.S. National Museum, University of Washing- (14C) 2-Deoxyglucoseuptake in the brain of the ton Burke Museum, the Tracy Aviary, and Charlie Ring Dove (Streptopeliarisoria). II. Differential Collins.The following individuals and organizations uptakeat the onsetof incubation.Brain Research provided field assistancein collectingspecimens: Sa- 700:137-141. rah A1-Tamimi,Jason Weckstein, Rob Moyle, Nigella GOODWIN,D. 1983. Pigeonsand Doves of the World, Hilgarth, Tony Jones,Aldolfo Navarro, A. Townsend 3rd ed. Cornell University Press, Ithaca, New Peterson, Mark Robbins, Moussa Barka, Imar Mous- York. sa, Wil Silkens, Arizona Fish and Game, Texas Parks HACKETT,S. J. 1996. Molecular phylogeneticsand and Wildlife, and the U.S. Fish and Wildlife Service. biogeographyof tanagersin the genusRampho- M. Griekspoorprovided soundrecordings. Dave En- celus(Aves). Molecular Phylogeneticsand Evo- strom, Larry Page, Fred Sheldon, and two anony- lution 5:368-382. 886 JOHNSONET AL. [Auk, Vol. 118

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