The Auk 117(1):22-32, 2000

MOLECULAR PHYLOGENY OF JACANAS AND ITS IMPLICATIONS FOR MORPHOLOGIC AND BIOGEOGRAPHIC EVOLUTION

LINDA A. WHITTINGHAM, TMFREDERICK H. SHELDON,2 AND STEPHENT. EMLEN3 1Departrnentof BiologicalSciences, University of Wisconsin-Milwaukee,Milwaukee, Wisconsin 53201, USA; 2Museumof Natural Science,119 FosterHall, LouisianaState University, Baton Rouge, Louisiana 70803, USA;and 3Sectionof Neurobiologyand Behavior, Division of BiologicalSciences, Cornell University, Ithaca, New York 14853, USA

ABSTRACT.--Wecompared sequences of mitochondrialcytochrome-b and ND5 genesin a phylogeneticanalysis of sevenspecies of jacanas,representing all six generaand including the GreaterPainted-snipe (Rostratula benghalensis) asan outgroup.When analyzedseparately by parsimonyand maximum-likelihoodbootstrapping, the two genesproduced consistent trees,although the ND5 tree was better resolvedthan the cytochrome-btree. When com- bined,the data from the two genesproduced a fully resolvedtree that was identicalto the ND5 tree.This tree had the followingform: ((((Irediparra,Microparra), Metopidius), Actophi- lornis),(( jacana, J. spinosa), Hydrophasianus) ), Rostratula. The phylogenyconsists of two majorclades that wereknown to traditionaland phylogenetictaxonomists. It alsocontains sistertaxa that are geographicallydisjunct: the New WorldJacana and Asian Hydrophasianus, and the AfricanMicroparra and AustralianIrediparra. We postulatethat this biogeographic pattern resultsfrom the extinctionof interveningAfrican and Asian taxa, respectively.Re- ceived31 May 1998,accepted 26 April 1999.

JACANASare worldwide inhabitantsof trop- Sibleyand Ahlquist1990, Chu 1995).Strauch's ical and subtropicalopen wetlands.Eight ex- (1978)character-compatibility (clique) analysis tant speciesin six generaare recognizedin the of 63 skeletal and seven muscular characters of family .Four genera are monotypic shorebirdsdefined two groupsof jacanagen- and occuron three continents:Microparra (Af- era: (1) Hydrophasianusand Jacana,and (2) Me- rica), Irediparra (), Hydrophasianus topidius,, Irediparra, and Micropar- (Asia),and Metopidius (Asia). Two othergenera ra. Following a critical appraisalof Strauch's consist of two species:Actophilornis africanus shorebirdstudy (Michevichand Parenti1980), () and A. albinucha(Madagascar), and Ja- Chu (1995)reanalyzed Strauch's data using cla- canajacana () and J. spinosa (Cen- distic parsimony.His reanalysisindicated the tral America).The generaof jacanasare mainly monophylyof jacanasbut was not designedto allopatric, but some co-occur in portions of resolverelationships within the family. Sibley Asia (Hydrophasianusand Metopidius)and Af- and Ahlquist's(1990) DNA-DNA hybridization rica (Actophilornisand Microparra).The two Ja- study indicatedthat Actophilorniswas much canaspecies co-occur and hybridizeonly in a closerto Irediparrathan to Jacana.Unfortunate- small area of western (Wetmore 1965). ly, Sibleyand Ahlquistdid not includeother ja- Until recently,relationships of the jacanasto canataxa in their comparisons. other groupsof were poorlyknown. It is To shedmore light on intergenericrelation- now clearthat they are membersof the Char- shipsin jacanas,we comparedDNA sequences adriiformesand probablyare most closelyre- of portionsof the mitochondrialcytochrome-b lated to painted-snipes,family Rostratulidae geneand the fifth subunitof nicotinamidead- (Kitto and Wilson 1966, Strauch1978, Sibley eninedinucleotide dehydrogenase gene (ND5) and Ahlquist1990). Within the Jacanidae,there of all sixgenera. These comparisons resolve the havebeen no thoroughmodern studies of in- jacanaphylogeny and providean initial assess- tergenericrelationships. The only systematic mentof the utility of ND5 sequencesfor study- studieshave been tangentsof larger efforts to ing avian phylogeny.Knowledge of the phy- reconstructshorebird phylogeny (Strauch 1978, logenyof jacanaspermits us to speculateon the evolutionaryforces that shapedtheir morphol- 4E-mail: [email protected] ogy and distribution.

22 January2000] JacanaPhylogeny 23

METHODS proportion of transversions(Moore and DeFillipis 1997); ti:tv was also estimated via the maximum- One individual from eachof sevenspecies repre- likelihoodoption in PAUP*(Swofford 1998). Phylo- sentingall six generaof the Jacanidaewere sampled genetic relationshipswere estimated using maxi- (Table 1): (Actophilornisafricanus), mum-likelihood (randomized heuristic search) and LesserJacana (Microparra capensis), Comb-crested Ja- maximum-parsimony(branch and bound search) cana (Irediparragallinacea), Pheasant-tailed Jacana methodswith PAUP*.Bootstrapping values (100 rep- (Hydrophasianuschirurgus), Bronze-winged Jacana licates) were determined for both maximum-likeli- (Metopidiusindicus), (Jacana jacana), hood and parsimonymethods to assesssupport for and NorthernJacana (J. spinosa). The GreaterPainted- internalbranches. Cytochrome-b and ND5 datawere snipe (Rostratulabenghalensis) was used as an out- combinedand analyzedseparately to observethe be- group becauseof its apparentclose relationship to haviorof the genes. the jacanas(Sibley and Ahlquist1990). Laboratorymethods.--In an attempt to avoid ampli- ficationof nuclear pseudogenes,we extractedDNA RESULTS from tissue,which has a relativelyhigh mitochon- drial DNA contentcompared with blood (Sorenson Characteristicsofsequence data.--We examined and Fleischer 1996, Quinn 1997). DNA was extracted a total of 705 nucleotides(345 of cytochromeb from 0.1 g of heart, liver, or muscletissue following and 360 of ND5) for sevenspecies of jacanas the standardphenol/chloroform extraction method and the outgroup(Rostratula). Average nucle- (Hillis et al. 1990).A portionof the mitochondrialcy- otide compositionsfor both genesand all po- tochrome-band ND5 geneswas isolatedand ampli- sitions were as follows: G = 11.3%, A = 30.2%, fied via the polymerasechain reaction (PCR). We used the following primers for cytochrome b T = 24.3% and C = 34.2%. Nucleotide compo- (H16065: GGAGTCTTCAGTCTCTGGTTTACAAGAC sitionalbias was highestat third-codonposi- and L15656: AACCTACTAGGAGACCCAGA; Helm- tions, intermediate at second positions, and Bychowskiand Cracraft 1993) and ND5 (H14149: lowestat first positions(Table 2). First positions CCTATTTTGCGGATGTCTTGTTC and L 13753: CAG- were biasedslightly, being G-poor and C-rich GAAAATCCGCTCAATTCGG; Garcia-Moreno et al. for cytochromeb andslightly T-poor and A-rich 1999). Primer sequencesare listed 5' to 3' and num- for ND5 (Table2). Secondpositions were G- bersrefer to the 3' baseof the primer referencedto poor and T-rich for both cytochromeb and the completemtDNA sequenceof the chicken(Gallus ND5. Third positionsof bothgenes were most gallus;Desjardins and Morais 1990).H and L refer to primerslocated on the heavyand light strandsof the heavily biasedtoward G- and T-poor and A- and C-rich. The total number of variable and mitochondrialgenome, respectively. PCR reactionswere done in 50-•L volumes using potentiallyphylogenetically informative sites 0.5 •M of eachprimer, 2.5 mM MgCI2,10 mM of each was similar for both cytochrome-band ND5 dNTP, and 3 U of Taqpolymerase. Thermal cycling fragments(Table 3). Althoughthe numberof began with 3.0 min at 94øCfor initial denaturation, variableamino acidsappeared lower for ND5 followedby 35 cyclesof denaturation(94øC, 1 min), than for cytochromeb, the numberof poten- annealing (46 to 50øC,1.5 min) and extension(72øC, tially informative amino acidswas similar (Ta- 1 min), and a final extensionof 10 min at 72øC.PCR ble 3). The numbersof variableand informative productswere run on a 2% low-meltagarose gel that was stained with ethidium bromide, allowing the siteswere highest at third- codonpositions, in- amplifiedDNA fragmentto be visualized under UV termediateat first positions,and lowest at sec- illumination and excisedfrom the gel. PCR products ond positions(Table 2). Thesetrends were ev- were purified and concentratedwith Wizard PCR identwhether or not the outgroupwas includ- PrepsKit (PromegaA7170) and then sequenceddi- ed with the jacanas. rectly on an ABI 373 Stretchautomated sequencer. The percentdivergence (using the Tamura Sequencesare deposited in GenBank under acces- and Nei [1993] correction)among taxa for cy- sion numbers AF146616 to AF146631. tochrome b and ND5 is summarized in Table 4 Dataanalysis.--Sequences were alignedby eye rel- and Figure 1. For cytochrome-bsequences, in- ativeto the chickensequence (Desjardins and Morals group divergenceranged from 2% (Jacanaja- 1990)using the editor and translatorin MEGA (Ku- mar et al. 1993).Base frequencies, variation, pairwise canato ]. spinosa)to 17.2%(Irediparra to ]acana transition and transversion values, and distances jacana). Similarly, ND5 ingroup divergence were determined with MEGA. Overall transition to ranged from 1.7% (Jacanajacana to J.spinosa) to transversionratio (ti:tv) was estimatedby plotting 17.4% (Microparrato Actophilornisand Micro- the pairwise proportion of transitionsversus the parra to Hydrophasianus).For the combined 24 WHITTINGHAM,SHELDON, AND EMLEN [Auk,Vol. 117

TABLE1. Distributionsand collectionlocalities of jacanaspecies sampled plus the outgroup.

Speciesa Distribution Samplenumber b Collectionlocality African Jacana Central and southern Africa LSUMNS B19187 Dallas Zoo (Actophilornisafricanus) Central and southeastern Africa LSUMNS B23806 (Microparracapensis) Comb-crested Jacana Malaysia, northeasternAustralia LSUMNS B14045 Australia Irediparragallinacea) Pheasant-tailed Jacana , Southeast Asia LSUMNS B14041 India (Hydrophasianuschirur- g•s) Bronze-wingedJacana India, Southeast Asia LSUMNS B14040 India (Metopidiusindicus) Mexico, Central America LSUMNS B28956 Mexico (Jacanaspinosa) Wattled Jacana Panama, northeastern South America LSUMNS B15237 (Jacanajacana) GreaterPainted-snipe Africa,India, SoutheastAsia 71 (C. Sibley) EasternMalaysia (Rostratulabenghalensis) Classificationfollows Sibley and Monroe (1990). LSUMNS = LouisianaState University Museum of Natural Science. data, ingroup sequence divergence ranged third-positiontransitions, if not transversions, from 2% (Jacanajacana to J. spinosa)to 16.5% often are saturatedabove the 10% divergence (Microparrato Actophilornisand Microparrato level (Mooreand DeFillipis 1997,Sheldon et al. Jacanajacana). It was not surprisingthat the 1999). It is difficult to tell the extent of satura- congenericNew World species(Jacana) exhib- tion in Figure2 becausedata on the x-axisand ited the lowest divergence.However, we did y-axis are not independent, and Tamura-Nei not expectto find the greatestsequence diver- distances(x-axis) can be foreshortenedby sat- gencebetween the two Africanspecies (Micro- uration, despite correction for back mutations parraand Actophilornis). The divergence pattern (Sheldonet al. 1999).Similarly, the ND5 data in Figure1 suggeststhat cytochromeb evolves (Figs.3A, B) do not appearsaturated, but again slightly faster,overall, than ND5 (17 out of 28 it is difficultto tell in theseplots. Moreover, we pointslie belowthe diagonal). cannotrely on the findingsof otherstudies us- To examinepatterns of nucleotidechange, we ing ND5 becausethere has been no compara- plotted pairwise proportionsof transitionsand tive work on ND5 in birds. To examinepoten- transversionsagainst overall distance(Tamura tial saturationof the data usingan alternative and Nei 1993).For cytochromeb, both first- and method,we plotted the proportionof transi- second-positiontransitions increased slightly tions versustransversions in Figure 4, as sug- with overall distance(Fig. 2A), whereasfirst- gestedby Moore and DeFillipis (1997).That and second-positiontransversions did not (Fig. plot indicatesthat the ti:tv ratio decreaseswith 2B). Third-positiontransitions (Fig. 2A) and increasingtransversion, suggesting that satu- transversions(Fig. 2B) increasedmore substan- ration slowsthe apparentrate of transitional tially with overalldistance than any otherpar- changeat higherlevels of divergence,at leastto tition. Of ND5 partitions,third-position tran- some degree. sitions(Fig. 3A) and transversions(Fig. 3B) in- Determining initial ti:tv was difficult be- creasedmost dramaticallyin relationto overall causeonly one pairwise comparison(Jacana ja- distance.ND5 first- and second-positiondiver- canawith J.spinosa) was outside the likelyzone gencedid not changemarkedly in relationto of saturation(on the left-handside of Fig. 4). overall distance(Figs. 3A, B). Thus,there were not enoughtaxa for a regres- Becausethe third-position transitionsand sion analysis.Moreover, the short sequence transversionsincreased steadily in comparison lengthsof the two genesmade it difficultto de- with overall geneticdivergence, these parti- terminean accurateratio evenfor the singleJa- tions appear generally unsaturated. However, cana-Jacanapair. There were no ND5 transver- other studies of cytochromeb indicate that sionsbetween them, and only three (synony- January2000] JacanaPhylogeny 25

TABLE3. Summary of variable sites and amino ac- ids in cytochromeb and ND5 for the sevenspecies of jacanas.Numbers in parenthesesinclude the outgroup (Rostratulabenghalensis).

Cyto- chrome b ND5 Total

Sites Total 345 360 705 No. variable 93 (102) 90 (101) 183 (203) No. informative 51 (59) 57 (67) 108 (126) Amino acids Total 115 120 235 No. variable 19 (23) 13 (14) 32 (37) No. informative 7 (7) 5 (5) 12 (12)

mous)cytochrome-b transversions. By includ- ing the otherfairly closelyrelated pairs of taxa (e.g.Jacana with Hydrophasianus),it was possi- ble to estimate approximateinitial ti:tv of 6:1 for ND5 and 3:1 for cytochromeb. The com- bined initial ti:tv appearsto be about3:1 (Fig. 4). Maximum-likelihoodsearches by PAUP*in- dicated a combined ti:tv of 3.48. One concernin this study was that because theDNA sequenceswere short, they might rep- resent accidentallyamplified nuclear pseudo- genes.However, the characteristicsof the se- quencesindicated that they were not pseudo- genes.The cytochrome-bsequences exhibited nucleotide composition, codon-position rates of change,and transitionand transversionpat- ternsthat are typical of the mitochondrial cytochromeb (e.g. Edwards et al. 1991, Nunn and Cracraft 1996, Slikas 1997). It is more dif- ficult to tell whether the ND5 sequencesare typicalbecause no publishedND5 datasets ex- ist for birds. However,the ND5 sequencesex- hibited patterns of base composition and changethat were very similar to the cyto- chrome-bsequences and showedno signs of pseudogenebehavior (Sorenson and Fleischer 1996, Quinn 1997).

PHYLOGENETIC ANALYSIS

Totalsequence data.--In the combinedanalysis of cytochrome-band ND5 nucleotidesequenc- es, maximum-parsimony (MP) and maximum- likelihood(ML) methodsproduced a singleto- pology (Fig. 5). The ML tree had a In of -2,437.1475, an estimated ti:tv at 3.48, and a site- specificrate factorfor the gamma distri- bution (alpha) of 0.17. WeightedMP analysis 26 WHITTINGHAM,SHELDON, AND EMLEN [Auk, Vol. 117

TABLE4. Percentdivergence (corrected for backmutations [Timura and Nei 1993])between taxa for cyto- chrome-b(above diagonal) and ND5 (belowdiagonal) sequences.

Species Jj lsp Hc Mi Aa Mc Ig Rb Jacanajacana -- 2.06 11.58 15.40 14.19 16.64 17.20 18.02 Jacanaspinosa 1.70 -- 11.21 15.05 14.54 16.64 16.88 18.21 Hydrophasianuschirurgus 9.75 10.43 -- 10.85 10.75 12.93 14.42 15.01 Metopidiusindicus 14.47 15.97 13.08 -- 15.28 12.94 14.85 16.17 Actophilornisafricanus 13.05 12.96 11.29 12.16 -- 15.75 15.01 17.41 Microparracapensis 16.48 14.95 17.36 14.68 17.35 -- 11.44 18.48 Irediparragallinacea 15.09 14.29 14.84 13.60 14.33 11.02 -- 18.98 Rostratulabenghalensis 14.51 14.45 15.30 17.05 14.13 22.29 17.27 --

(ti:tv = 3) produceda single most-parsimoni- oustree (length= 556 steps,CI excludingun- informativecharacters = 0.734).Bootstrap val- ues (100 replicates)ranged from 72 to 100%for MLand 66 to 100% for MP (Fig. 5). In the ML/ 0.25 MP tree, two major cladeswere evident.The first consistedof four taxa:((Irediparra, Micro- 0.20- parra), Metopidius),Actophilornis. The second A AA•I• A clade consistedof the two New World species, •= 0.15 A A Jacanajacana and J. spinosa, plus Hydrophasianus :• astheir sister group. • 0.10 Cytochrorne-bdata.--The analysis of cyto- • chrome-bsequences alone (Fig. 6) resulted in a • 0.05 singleMP tree (length= 281 steps,CI exclud- ing uninformativecharacters = 0.758,weight- ed transition:transversion= 3), which was 0.00 identical to the ML tree (In = -1,203.9366, I I I I PAUP*estimated ti:tv at 3.46, and the site-spe- 0.00 0.04 0.08 0.12 0.16 0.20 cific rate factor for the gamma distributionat 0.19). When bootstrapped,the cytochrome-b 0.25 tree indicated a very strong associationbe- tween Jacanajacana and J.spinosa, and between 0.20- Irediparraand Microparra. In thisrespect, it was o 0.15 similar to the total sequencetree. However the ,_ A A AAAA 0 25 •= O.lO

A • 020 • 0.05 A • ß O

ß• 045 . 0.00 Z ß jee ß -

• 010 -0.05

0.00 0.04 0.08 0,12 0.16 0.20 005 Z Cytochromeb Tamura-NeiDistance

0 00 , , 000 oo5 040 045 o2o o25 FIG. 2. Divergenceof cytochrome-btransitions (A) and transversions(B) relative to corrected dis- Cytochromeb Tamura-Nei Distance tance (Tamura and Nei 1993) at first- (circles), sec- F•, 1, Divergenceof cytochromeb versusND5 ond- (squares),and third-codon (triangles)posi- for jacanasand the Rostratulaoutgroup. tions. January2000] JacanaPhylogeny 27

0.35 • 014 A ß 0.30 -'• 0 12 ß r- • 010 •n 0.25 I ß ß ß • 0 08 r- ß Iii i II ii I -• 0.20 ß .--• 0 06

I-- 0.15 __ • 0 04 Q .o z O.lO OQ_002 0 0.05 Q' 0 O0 0 00 0 01 0 02 0 03 0 04 0 05 0 06 0 07 0.00 Proportional Distance -- Transversions

I I i r FIG. 4. Plot of transitions versus transversions for 0.00 0.05 0.10 0.15 0.20 0.25 combinedcytochrome-b and ND5 sequences.

0.35 -

0.30 - tophilornis,Metopidius, and Hydrophasianusin • 0.25- ._ the cytochrome-btree.

m>0.20 - DISCUSSION

I-- 0,15 - && ß Our analysis of cytochrome-band ND5 se- z O,lO - quencedata produced a fully resolvedphylog- eny indicatingthe divisionof the Jacanidaeinto 0.05 - two clades:(1) Australian Irediparra,African Microparra,Asian Metopidius,African Actophi- o,oo -

I I I lornis,and (2) Asian Hydrophasianusand New 0.00 0.05 0.10 0.15 0.20 0.25 WorldJacana. The phy!ogenyof thesetaxa is as follows:(((Irediparra, Microparra), Metopidius), ND5 Tarnura-Nei Distance Actophilornis),((Jacana jacana, J. spinosa), Hydro- FIG. 3. Divergence of ND5 transitions (A) and phasianus). transversions(B) relative to corrected distance (Ta- In general,previous classifications of jacanas mura and Nei 1993) at first- (circles), second- havebeen consistent with thisphylogeny. Wag- (squares)and third-codon(triangles) positions. ler (1832in Johnsgard1981) recognized seven speciesof jacanasand placed them in three genera:Jacana, Hydrophasianus, and Metopidius. positionsof Actophilornisvis-a-vis Metopidius Jacanaand Hydrophasianus were monotypic gen- was unclear,as was the positionof Hydrophas- era and thoughtto be eachothers' closest rel- ianusin regardto theJacana species. Compared atives.Metopidius included the five speciesof with the total sequencetree and the ND5 tree Old World"typical" jacanas: capensis, africanus, (seebelow), the cytochrome-btree was poorly albinucha,gallinacea, and indicus.In classifica- resolved. tionssubsequent to Wagler's,the five "typical" ND5 data.--Analysisof ND5 sequencesusing Old Worldjacanas were split into fourseparate, bothMP and ML produceda singletree (Fig. and mostly monotypic, genera (e.g. Peters 5) identical to the total sequencetree (MP: 1934):Microparra, Actophilornis, Irediparra galli- length= 296steps, CI excludinguninformative nacea,and Metopidiusindicus. characters= 0.725, weightedti:tv = 3; In = Previousphylogenetic studies also produced - 1,227.2669, PAUP* estimated ti:tv at 3.78, and results largely consistentwith ours. Strauch's the site-specificrate factorfor the gammadis- (1976,1978) character-compatibility analysis of tribution [alpha] at 0.15). The cytochrome-b, Charadriiformesrevealed two groupsof jaca- ND5, and combinedtrees were congruent,giv- nas correspondingto thosethat we found.One en the low bootstrapvalues for positionsof Ac- included Hydrophasianusand Jacana,and the 28 WHITTINGHAM,SHELDON, AND EMLEN [Auk,Vol. 117

0.080 Microparra 0.054 98/99 0.069 0.031 Irediparra 92/76

0.092 0.025 Metopidius 79/80

0.073 Actophilornis

0.008

lOO/lOO0.080rIo.o12 Jacanajacana 0.017 Jacanaspinosa 0.158 72/66

0.042 Hydrophasianus

Rostratula FIG.5. Phylogeneticrelationships among the Jacanidaebased on maximum-likelihood(ML) and maxi- mum-parsimony(Mr) analysesof 345 nucleotidesof cytochromeb and 360 nucleotidesof ND5. Identical branchingpatterns were determined by both analyses (Mr singlemost- parsimonious tree length = 556steps, CI excludinguninformative characters = 0.734;ML tree In = -2,437.1475). See text for more details. Per- centagebootstrap support from both methods of phylogeneticanalysis is shownto theleft of internalbranch- es (ML / MP).

other includedMicroparra, Irediparra, Metopi- found in Metopidius(Strauch 1976). This led dius,and Actophilornis.The Hydrophasianus-Ja-Strauch (1976) to placeMetopidius basal to Ire- cana group was defined by shared-derived diparra,Microparra, and Actophilornis.In con- wing spursand the othergroup (the "typical" trast, our molecular analysisplaces Actophilor- Old World species)by shared-derivedwing nis basal to Irediparra,Microparra, and Metopi- knobs and a blade-like radius. These two dius.Based on his results,Strauch (1976) rec- groups also are distinguishedby plumage ommended recognizing only two genera of characters(Strauch 1976). Both Hydrophasianus jacanas: Actophilornis (including Actophilornis, andJacana have light-colored wing stripesin all Irediparra,Microparra, and Metopidius),and Ja- plumages(except for neonatalplumage), and a cana(including ]acana and Hydrophasianus). blackeye stripe is presentin youngbirds. None Strauch's (1976, 1978) method--character of the othergenera of jacanas has either of these compatibilityor cliqueanalysis--was criticized characterstates in any plumage. However, by Mickevichand Parenti(1980), who preferred within the"typical" Old Worldgroup, the Aus- a parsimonyapproach. However, a recentre- tralian and two African genera(Irediparra, Mi- analysisof Strauch'sdata using modern cladis- croparra,and Actophilornis)share a derived tic parsimony failed to resolve any relation- character(fusion of the ischiumand pubis)not shipswithin the Jacanidae(Chu 1995).In con- January2000] JacanaPhylogeny 29

Cytochrome b ND 5

0.076 Microparra 0.088 Microparra 0.056 0.052 94/96 0.079 82/83 Irediparra 0.060 0-011r_I 0.055 Irediparra 59/591I 0.086 79/88 Actophilornis 0.083 Metopidius

0.017 0.104 Metopidius 0.056 <50/52 ,4ctophilornis

I 0.037 Hydrophasianus 0.003

0.011 I 100/1000.056[0.014[Jacanajacana 0.097 10.026 Jacanaspinosa 100/100 •0.Jacana jacana 011 84/85 0.177 Jacanaspinosa I 0.049 0.13 Hydrophasianus

Rostratula Rostratula FIG.6. Phylogeneticrelationships among the Jacanidaebased independently on the analysisof cyto- chromeb (345nucleotides; MP singlemost-parsimonious tree length = 281steps, CI excludinguniformative characters;0.758; ML treeIn = - 1,203.9366)and ND5 (360nucleotides; MP singlemost-parsimonious tree length= 296 steps,CI excludinguniformative characters = 0.725;ML tree In = -1,227.2669)mitochondrial genes.For details,see text. Percentagebootstrap support froin both inethodsof phylogeneticanalysis is shown to the left of internal bran&es (ML/MP).

trast,the tree producedby cliqueanalysis sub- genera:Jacana (for Jacana,Actophilornis, Metopi- stantially resemblesour molecular tree. So, at dius,and Irediparra),Hydrophasianus, and Mi- least for the Jacanidae,if not for the Charadri- croparra.The main reasonhe lumped the first iformes,clique analysis appears to be an effec- four generainto Jacanawas that they are "less tive approach. heterogeneous"than other groupsof shore- Sibleyand Ahlquist(1990) compared three birds that havebeen lumped (e.g. lapwings). jacanagenera by DNA-DNA hybridization(Ire- He keptHydrophasianus chirurgus and Micropar- diparra,Jacana, and Actophilornis) using Iredipar- ra capensisin their own generabecause of the ra as the radio-labeled control and Rostratula as distinctivenessof thesetwo species.Such an the radio-labeledoutgroup. The pairwisedis- approachdoes not meet modern (cladistic) tances in their tree were AT50H 4.9 for Iredi- standards.It groupstaxa by overallsimilarity parra-Actophilornis(three comparisons)and and ignores potential information in shared- AT50H 8.0 for Irediparra-Jacana(two compari- derived characters(e.g. wing spursfor Jacana sons).Their findingsare consistentwith ours and Hydrophasianus).Nevertheless, Fry's sum- and Strauch's(1976, 1978), and their data also mary of jacanacharacters contains several in- suggestthat a substantialdivergence gap sep- terestingobservations about their evolution.He aratesthe two main groupsof jacanas. observedthat only Actophilornis,Metopidius, Fry's (1983) study appearsat first to contra- andIrediparra share a uniqueblade-like radius, dict ours.Using an evolutionary-taxonomyap- whichpresumably is an adaptationfor wing- proach,he examinedvarious characteristics of broodingof eggsand downy young,whereas thejacanas and concludedthat there were three Jacana,Hydrophasianus, and Microparrahave a 30 WHITTINGHAM,SHELDON, AND EMLEN [Auk, Vol. 117 typical avian radius. He also noted that the and Asia). The New World-Asian(Jacana-Hy- plumageof adult Microparrais similarto that drophasianus)distribution is similarto thestork of juvenilesin the otherspecies. The only other distributionwithout an Africanrepresentative. adultjacana to havesuch plumage is the non- Furthermore,the African-Australian(Micropar- breedingHydrophasianus. From theseobserva- ra-Irediparra)distribution is similarto thestork tions,and others(e.g. patterns of radiusdevel- distribution without an Asian or New World opmentin juveniles),he concludedthat Micro- representative.Quite likely, the jacanadistri- parrais neotenic,having retained the juvenile butions are relicts of wider distributions in radiusand plumage.This conclusion makes ex- which the interveningtaxa becameextinct. cellentsense and also providesan explanation Sucha patternapplies to severalnonpasserine for homoplasyin jacanamorphology. For ex- groups.For example,the tiger-,which ample,the "typical" avianradius described by are representedby three speciesin the New Fry (1983)is a synapomorphyuniting Jacana World (Tigrisoma),one in Africa (Tigriornis), and Hydrophasianusand a convergencedue to and one in New Guinea (Zonerodius),would be neotenyin Microparra.Similarly, the retention an exampleof a group that may have under- of juvenilecharacter states also explains how gone extinctionin Asia. Microparraobtained its plumageand the pos- This patternof extinctionof interveningspe- siblecauses of the uniquenonbreeding plum- cies is most obvious in depauperategroups, age of Hydrophasianusand the appearanceand suchas jacanas, but it occursin speciosegroups disappearanceof frontal shieldsin variousspe- aswell. Wehave found, for example,closely re- cies. lated sisterspecies of swallows in Africa (Pseu- Biogeography.--Thejacanas have a pantropi- dhirundogriseopyga) and Australia (Cheramoeca caldistribution. Like other groups of birdswith leucosternus)without interveningAsian rela- similar distributions,such as storks,tiger-her- tives (Sheldonand Winkler 1993). Moreover, ons, finfoots, trogons, and barbets (Houde thispattern seems to applymainly to relatively 1994, Lanyon and Hall 1994, Houde et al. 1995, vagile open-countryor marsh-dwellingtaxa, Sheldonet al. 1995,Slikas 1997), jacanas are too suchas jacanas, storks, herons, and swallows. youngto be Gondwanan(Sibley and Ahlquist Thesegroups have redundant distributions, in- 1990)but are old enoughto havedeveloped id- dicating multiple invasionsof different conti- iosyncraticbiogeographic patterns. Thus, there nents.The forest-dwellinggroups, such as tro- is no simplegeographic scenario (e.g. common gons and barbets, with monophyleticNew vicariancepattern or dispersalroute) to explain World representatives(Sibley and Ahlquist how jacanasand otherpantropical bird groups 1990, Lanyon and Hall 1994, Espinosade los became distributed in the manner we observe Monteros 1998), have largely nonredundant today.However, given new phylogeneticinfor- distributions.These distributions suggest these mation, a few recurrent patterns among some groups have moved unidirectionallyaround pantropical groups are beginning to emerge. the globe.The differencein dispersalpatterns These patternsare enoughto suggestsome amonggroups suggests that we mustfactor be- large-scaleprocesses behind currentdistribu- havior (e.g. dispersalpotential), ecology (e.g. tions. For example,two of the more unusual habitat preferences),and geography (e.g. phylogeneticrelationships among the jacana routesof dispersal),at the very least,into a generaare the sistergroupings of (1) the New general discussionof patterns and causesof WorldJacana and the AsianHydrophasianus and pantropicalbird distribution. (2) the AfricanMicroparra and AustralianIre- diparra.These distributions are reminiscentof ACKNOWLEDGMENTS the distributions of other groups, such as storks.We knowthat theJabiru (Jabiru mycteria) We thank Tamar Cassidy(Transvaal Museum), S. of the New World is mostclosely related to the R. Grubh (Institute for Restorationof Natural Envi- ronment, India), Patricia Escalante (University of Saddlebill (Ephippiorhynchussenegalensis) of Af- Mexico), LSU Museum of Natural Science, Stuart rica and the Black-neckedStork (E. asiaticus)of Butchart,and the late CharlesSibley for donatingtis- Asia and Australia (Slikas1997). 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