Genetic and morphological relationships of the Berlenga wall lizard "Podarcis bocagei berlengensis] Lacertidae#

Transcription

1 J[ Zool[ Syst[ Evol[ Research 27 "1999# 84Ð091 Received on 2 September 0888 Þ 1999 Blackwell Wissenschafts!Verlag\ Berlin ISSN 9836! Centro de Ecolo`ia Aplicada\ Universidade de E vora\ E vora and\ 1 Centro de Biolo`ia Ambiental\ Universidade de Lisboa\ Lisboa\ Portu`al Genetic and morphological relationships of the Berlenga wall lizard "Podarcis bocagei berlengensis] Lacertidae# P[ SAł!SOUSA 0\1 \A[P[ALMEIDA 1 \H[ROSA 1 \L[VICENTE 1 and E[ G[ CRESPO 1 Abstract Six biometric characters and 04 electrophoretic loci of three known subspecies of Podarcis boca`ei were studied[ Contrary to biometric indications\ Podarcis boca`ei berlen`ensis showed a closer genetic relationship with Podarcis boca`ei carbonelli than with the nominal subspecies[ The biometric results were confounded by the relative large size of P[ b[ berlen`ensis[ Post!glacial colonization from two di}erent areas might explain the greater genetic di}erentiation between the P[ b[ boca`ei and P[ b[ carbonelli populations "Nei s D 9[047 near the species threshold#[ It might also explain the known colonization of these two lizards through the islands o} the west coast of the Iberian penninsula and their present distributions to the north of "Galicia^ Northern Portugal# and to the south of the valley of river Douro "central Portugal\ western and Sistema Central#\ respectively[ Key words] Wall lizard Ð speciation Ð biometric data Ð allozymes Introduction The Bocage wall lizard Podarcis boca`ei Seoane 0773 occurs where Atlantic!type conditions prevail[ It is an IberianÐAtlantic endemic species that probably originated in the north!western corner of Iberia "Galan 0875\0886^ Perez!Mellado 0886#[ It is a ground!dwelling\ insectivorous lizard\ whose males show dark green\ reticulated dorsal patterns\ whereas the females are brown with a pair of green stripes "Perez!Mellado 0870a\ 0875#[ The systematics of the lizard have been revised taking only its morphology into account "e[g[ Perez!Mellado 0870b^ Galan 0874\ 0875^ Perez!Mellado and Galindo 0875#[ Until now three subspecies have been recognized "see Table 0#[ First\ the nom! inal subspecies Podarcis boca`ei boca`ei is distributed in north! western Iberia\ e[g[ western Asturias\ Cantabria\ Galicia and the north of Portugal\ where it occurs in sympatry with the Iberian wall lizard Podarcis hispanicus "Galan 0875^ Sa!Sousa 0887#[ Second\ there is the subspecies Podarcis boca`ei car! bonelli Perez!Mellado 0870 which was initially thought to be restricted to the mountain range of western {Sistema Central "Perez!Mellado 0870b\ 0872\ 0873\ 0875#[ However\ P[ b[ car! bonelli\ has also been found in other mountains and along the Atlantic coastal lowlands\ particularly in Portugal "Magraner 0875^ Sa!Sousa 0884\ 0888#[ Third\ there is the insular subspe! cies\ Podarcis boca`ei berlen`ensis Vicente 0874\ which is described as a relative large form that is related to the popu! lations of P[ boca`ei of NW Spain "Galicia#\ and is restricted to the Berlengas islands o} the western coast of Portugal "Vicente 0874#[ This subspecies has been also found near the coast in the adjacent "pen#insula of Peniche "Sa!Sousa 0881#\ a fact that is consistent with the low genetic di}erentiation found between lizards from Berlenga and from Peniche "Almeida et al[ "0881#[ Electrophoretic investigations of various Lacertidae\ have frequently suggested a reduction in the number of subspecies "Cirer and Guillaume 0875^ Ramon et al[ 0880#\ and clari_ed the relationships within some complex of species "Fu et al[ 0884^ MacCulloch et al[ 0884^ Bobyn et al[ 0885#[ This technique can also be used to calculate estimates for evolutionary divergence "Gorman et al[ 0864^ Guillaume and Lanza 0871^ Mayer and Tiedemann 0871^ Busack 0875# or to demonstrate that some populations are less related to each other than previously sup! posed "Guillaume et al[ 0865^ Busack 0876^ Capula 0883^ Mayer and Arribas 0885#[ Guillaume and Cirer "0874# suggested that Nei s genetic dis! tances "identities# of a magnitude of D ¾ 9[1 "I 9[7# are com! patible with an intraspeci_c di}erentiation\ whereas 9[2 ¾ D ¾ 9[4 "9[64 ¾ I ¾ 9[59# indicates congeneric species\ and D 9[8 "I ¾ 9[3# implies that species belong to di}erent genera[ Guillaume "0878# generated a chronological calibration of D for lacertids estimating that Nei s D 0[9 is equivalent to 09Ð02 My ago\ taking into account Busack s "0875# data and the age of the opening of the Strait of Gibraltar at 4[4Ð6[9 My ago[ We studied the di}erentiation of P[ b[ berlen`ensis in relation with the other two subspecies of P[ boca`ei\ both from a mor! phological point of view using biometric data and from a genetic point of view using allozyme electrophoresis[ Materials and methods We collected P[ boca`ei samples from four Portuguese sites within the distribution range of the three known subspecies of this lizard "Fig[ 0#[ P[ b[ boca`ei was from Gere¼s "G#\ a mountain region in the North of Portugal^ P[ b[ berlen`ensis came from the island of Berlenga Grande "B# Ð hereafter referred as Berlenga Ð and from the adjacent coastal area of Peniche "P# and _nally\ P[ b[ carbonelli came from Cabo Raso "C#\ a coastal region south of the Berlengas islands[ Six biometric characters were measured] SVL snout!vent length\ HL head "pileous# length\ HW head width\ HH head height\ FLL forelimb length "from the axillar ring to the end of the fourth _nger# and HLL hindlimb length "from the groin ring to the end of the fourth toe#[ Keeping the sexes apart\ two di}erent statistical analyses allowed the evaluation of biometric data from the samples[ Principal component analyses "PCA# were extracted from a standardized product!moment correlated matrix[ Squared Mahalanobis distance "MahD1# was also used to obtain biometric matrices\ since MahD1 takes the correlations among variables into account and is independent of their scales "Leg! endre and Legendre 0887#[ The UPGMA method was used for cluster analysis[ Descriptive statistics of samples are shown in Appendix 0[ By allozyme electrophoresis\ we examined a total of 04 presumptive loci] 02 loci\ encoding 00 enzymes were investigated using 01) starch gel electrophoresis "standard horizontal method#\ and two other loci "albumin and haemoglobin# were scored through cellulose acetate gel electrophoresis[ Both techniques used are described by Pasteur et al[ "0876# and Rosa and Oliveira "0883#[ All protein systems were analysed U[S[ Copyright Clearance Center Code Statement] 9836Ð4634:99:2791Ð9984,04[99:9

2 85 SAł!SOUSA\ ALMEIDA\ ROSA\ VICENTE and CRESPO Table 0[ External characters that distinguish lizards from the three subspecies of Podarcis boca`ei Characters Podarcis boca`ei boca`ei Podarcis boca`ei carbonelli Podarcis boca`ei berlen`ensis Sex m f m f m f Body size great moderate small to small great great moderate Head depth robust moderate robust moderate robust moderate robust robust Dorsal pattern striped thin reticulated^ striped striped discontinuous^ striped striped dis:continuous discontinuous^ striped thick continuous discontinuous reticulated dis:continuous reticulated Dark\ ventral intense moderate low low intense intense pigmentation 3Ð5 rows 1Ð3 rows 9Ð1 rows 9Ð1 rows 3Ð5 rows 3Ð5 rows Dorsal colour intense green brown^ green^ brown^ intense green brown^ 0 ocreous brownish ocreous ocreous 1 Light stripes green green green green^ green green yellowish Flank color hazel!brown hazel!brown intense green brown^ intense green brown^ ocreous ocreous Belly color yellow^ yellow^ white white^ white^ white orange ocreous pale yellow nacreous Blue spots no no yes yes yes yes many moderate many moderate during the mating period[ Fig[ 0[ Known Iberian distribution of the three subspecies of Podarcis boca`ei] nominal subspecies "light grey shaded#\ P[ b[ carbonelli "dark grey shaded and border# and P[ b[ berlen`ensis "ampli_ed window#

3 Genetic and morphological relationships of Berlenga wall lizard 86 Fig[ 1[ Graphs of the projections of biometric data from males and females of Podarcis boca`ei on the two _rst principal components "PC0 PC1 explained variance)#] males "66[94 03[39 80[34)#\ females "56[06 06[72 72[99)# using material from the liver\ skeletal muscle\ heart or blood\ depending on the best results achieved in a pilot study[ The enzymes score\ the tissues used for the respective enzyme\ and electrophoretic conditions applied are presented in Appendix 1[ For each protein system encoded by two or more presumptive loci\ the isozymes were assigned by num! bers starting from the allozyme band with the greatest anodal migration[ Alleles of each locus were indicated by numbers proportional to the relative migration of their allozymes\ using {099 for the most frequent allele[ Genetic variability was measured through mean heterozygosity "unbiased estimate\ Nei 0867# and percentage of polymorphic loci\ whereas the genetic di}erentiation was quanti_ed through Nei s "0867# unbiased index D\ from which UPGMA phenograms were constructed[ Results The ordination of centroids of the male and female locality samples of P[ boca`ei in the space of the _rst two principal components of variation "PC0\ PC1#\ deduced from the six biometric characters\ is shown in Fig[ 1[ The associated factor loadings and cumulative percentages of variance they explain are shown in Table 1[ PC0 "66Ð72)# re~ects general {size and it is positively "and signi_cantly 9[69# associated with all other characters[ PC1 "03Ð07)# re~ects intersample variation within P[ boca`ei] a clear separation occurs only for females of P[ b[ carbonelli from the other subspecies\ with a large overlap of individual biometric projections\ particularly within males "Fig[ 1#[ Squared Mahalanobis distances are shown in Table 2 and their UPGMA phenograms are depicted in Fig[ 2[ Biometric data showed that males of P[ b[ boca`ei and of P[ b[ berlen`ensis from Berlenga were similar and bigger than those from Pen! iche:cabo Raso[ For females\ there were no signi_cant di}er! ences within P[ b[ berlen`ensis "Berlenga and Peniche#\ but there was a biometric similarity between the samples of P[ b[ boca`ei and P[ b[ berlen`ensis[ Of the 04 loci analysed\ seven "36)# were monomorphic in all samples] AAT!1\ AK\ CK\ LDH!1\ MDH!0\ PEP\ SOD[ For the remaining eight loci\ at least two di}erent alleles were detected per locus[ The respective frequencies are presented in Table 3[ No locus was discriminative among the four samples[ EST!1 was strongly polymorphic\ whereas two loci "MPI\ PGM!1# were almost dimorphic in all samples[ For two of the polymorphic loci\ AAT!0 and PGD\ there were two pre! dominant alleles "AAT!009\ PGD099# shared by P[ b[ berlen! `ensis "Berlenga and Peniche# and P[ b[ carbonelli "Cabo Raso#\ whereas the other two alleles "AAT!049\ PGD029# were the most frequent in P[ b[ boca`ei "Gere¼s#[ For the loci LDH!0\ ALB and HB\ the alleles predominating in P[ b[ berlen`ensis "LDH!0099\ ALB099\ HB099# were di}erent from those that occurred most frequent in the mainland subspecies\ P[ b[ boca`ei "Gere¼s# and P[ b[ carbonelli "Cabo Raso#[ Unique rare alleles occurred at Cabo Raso "AAT!0022\ EST! 142#\ Gere¼s "PGD059# and Peniche "PGM!1028#[ Conformity to HardyÐWeinberg equilibrium expectations was found for all loci\ except for the locus MPI "Cabo Raso and Peniche#\ for which a reduced number of heterozygotes "chi!square\ p 9[94# was obtained[ Genetic variability measures are shown in Table 4[ Mean heterozygosity was greatest for mainland sub! species P[ b[ boca`ei "Gere¼s# and P[ b[ carbonelli "Cabo Raso# whereas a lower heterozygosity was found for the P[ b[ berlen! `ensis "Berlenga and Peniche#[ Polymorphism was 0[4 to 1 times greater in mainland samples "Gere¼s^ Cabo Raso# than in the insular ones\ using both 88 and 84) criteria[ Genetic distances are shown in Table 5 and the UPGMA phenograms deduced are shown in Figure 2[ Within P[ b[ berlen`ensis "Berlenga and Peniche# there was low di}erentiation "Nei s D 9[997Ð9[900#[ Following Guillaume and Cirer "0874# criteria\ P[ b[ carbonelli "Cabo Raso# is linked either with P[ b[ berlen`ensis on a sub! speci_c genetic level "Nei s D 9[000# or with P[ b[ boca`ei from Gere¼s "Nei s D 9[047#[ Finally\ P[ b[ berlen`ensis reached the species threshold in relations with P[ b[ boca`ei "Nei s D 9[134Ð9[149#[ Discussion Biometric data "PCA\ MahD1 clusters# mask the inferred closer genetic relationship between P[ b[ berlen`ensis and P[ b[ carbonelli\ although the morphological resemblance based on external characters "e[g[ chromatic patterns\ see Table 0# agrees with the genetic data[ In contrast to P[ b[ boca`ei\ both P[ b[ berlen`ensis and P[ b[ carbonelli show individuals with blue spots on the outer ventral scales and a white belly^ the males always have green ~anks independently of the greenish to brownish coloration of the mid!back and they share similar thin reticulated patterns^ with the exception of the trait {size \ females of both forms also resemble each other in their chro! matic patterns "Table 0#[ Thus\ the biometric clusters found

4 87 SAł!SOUSA\ ALMEIDA\ ROSA\ VICENTE and CRESPO Table 1[ Factor loadings and cumu! lative percents of explained vari! ance for the three _rst principal components of six biometric characters measured among four samples of Podarcis boca`ei[ Bio! metric characters are described in the text Males Females n093 n52 Variable PC0 PC1 PC2 PC0 PC1 PC2 SVL 9[8549 9[9647 9[9274 9[7581 9[9063 9[3372 HL 9[8484 9[9700 9[9572 9[8937 9[0840 9[1424 HW 9[7654 9[1735 9[2307 9[7022 9[2755 9[0707 HD 9[5780 9[5506 9[1775 9[6333 9[3521 9[2404 FLL 9[7712 9[2625 9[0524 9[5822 9[5263 9[1047 HLL 9[7547 9[3284 9[9571 9[6887 9[4003 9[9638 cum[ ) expl[ variance 66[94 80[34 84[30 54[06 72[99 89[89 PCA loadings extracted from product!moment correlation matrices[ Table 2[ Squared Mahalanobis dis! tance matrix for four samples of Podarcis boca`ei "males above diag! onal\ females below diagonal# not signi_cantly di}erent at p ³ 9[94 Subspecies Sample G B P C P[ b[ boca`ei Gere¼s Ð 4[973 00[546 12[887 P[ b[ berlen`ensis Berlenga 3[711 Ð 7[259 05[468 P[ b[ berlen`ensis Peniche 1[675 0[575 Ð 1[890 P[ b[ carbonelli Cabo Raso 03[078 00[927 09[416 Ð Fig[ 2[ Distance phenograms resulting from UPGMA clustering based on squared Mahalanobis distance "morphology# and Nei s "0867# unbiased distance "electrophoresis#[ Letters at the end of branches cor! respond to abbreviations of sampling sites "see legend in Tables 2\3\4\5#[ r cof[ is the cophenetic correlation illustrate the relative large size of P[ b[ berlen`ensis\ not in comparison with P[ b[ boca`ei\ but with P[ b[ carbonelli "see genetic data below#[ Lizard gigantism is a phenomenon that is widespread among insular populations of Podarcis\ e[g[ in Podarcis lilfordi\ Podarcis muralis\ Podarcis pityusensis\ Pod! arcis tili`uertus "Cirer 0870\ 0886^ Salvador 0875^ Cheylan 0877^ Perez!Mellado and Salvador 0877^ Delauguerre and Cheylan 0881#[ Lizards tend to show a size increase and a greater mor! phological homogeneity "by genetic drift# on smaller and older islands "Cheylan 0877^ Cirer and Mart(nez!Rica 0889#[ The reduction of predator pressure and the simultaneous occurrence of high population density of insular lizards leads to a shortage of resources and the increase of intraspeci_c competition and sexual dimorphism linked with agonistic behaviour of males\ thus creating a trend towards gigantism "Thorpe 0874^ Cheylan 0877^ Vicente 0878^ Cirer and Mart(nez!Rica 0889^ Case and Schwaner 0882#[ Conversely\ we found less overlap of biometric characters in all females\ probably re~ecting indirectly the selec! tive pattern that prevails in the males of P[ b[ berlen`ensis "Fig[ 1#[ Both\ the results of Almeida et al[ "0881# and of this study show that genetic di}erentiation among lizards from the Berlen! gas islands "e[g[ Berlenga\ Estelas\ Farilha½o Grande# and from Peniche are low "Nei s D 9[997Ð9[935#[ This is in accordance with their taxonomic status pertaining to P[ b[ berlen`ensis[ However\ we also found an accidental introduction of P[ b[ berlen`ensis from Berlenga into Peniche harbour and its sur! roundings[ Thus\ a process of intergradation may occur there between the introduced lizards and those from the aboriginal population of Peniche[ The greater value of genetic distance "Nei s D 9[047# between P[ b[ carbonelli and P[ b[ boca`ei probably means that these two lizards subspecies evolved in two di}erent refugia during glaciations[ We and other authors presume that the great morphological homogeneity reported within mainland populations of P[ b[ boca`ei may re~ect their recent postglacial expansion from a north!west Galician refugium "Galan 0875^ Perez!Mellado and Galindo 0875^ Perez!Mellado 0886#[ Thus\ former populations of P[ boca`ei might have occupied western Iberia more or less extensively until the climatic changes of

5 Genetic and morphological relationships of Berlenga wall lizard 88 Table 3[ Allelic frequencies cal! culated for all 04 presumptive loci in each sample P[b[b P[b[be P[b[c Gere¼s Berlenga Peniche Cabo Raso Locus Allele G B P C AAT!0 022 Ð Ð Ð 9[ [099 0[999 0[999 9[ [899 Ð Ð Ð n AAT! [999 0[999 0[999 0[999 n AK 099 0[999 0[999 0[999 0[999 n CK 099 0[999 0[999 0[999 0[999 n EST! [310 Ð Ð 9[ [468 9[464 9[126 9[ Ð 9[314 9[652 9[ Ð Ð Ð 9[980 LDH! [614 9[249 9[314 9[ [164 9[549 9[464 9[025 LDH! [999 0[999 0[999 0[999 MDH! [999 0[999 0[999 0[999 n MPI 000 Ð Ð Ð 9[ [864 0[999 9[836 9[ [914 Ð 9[942 Ð n PEP 099 0[999 0[999 0[999 0[999 n PGD 059 9[945 Ð Ð Ð 029 9[833 Ð Ð 9[ Ð 0[999 0[999 9[753 PGM!1 028 Ð Ð 9[914 Ð 099 0[999 0[999 9[864 0[999 SOD 099 0[999 0[999 0[999 0[999 n ALB 091 9[472 Ð Ð 9[ [306 0[999 0[999 9[399 n HB 099 9[021 9[623 9[699 9[ [757 9[155 2[999 9[849 n number of individuals scored successfully for each locus[ Table 4[ Genetic variability mea! sures at 04 presumptive loci in all samples Subspecies Sample H P 88) P 84) A P[ b[ boca`ei Gere¼s G 9[015 39[9 39[9 0[3 P[ b[ berlen`ensis Berlenga B 9[979 19[9 19[9 0[1 P[ b[ berlen`ensis Peniche P 9[984 22[2 15[6 0[2 P[ b[ carbonelli Cabo Raso C 9[016 35[6 39[9 0[5 H mean heterozigosity\ unbiased estimate^ A mean number of alleles per locus[ P 88)\ P 84) percentage of polymorphic loci\ 88) and 84) criteria[ Pleistocene\ an event that provoked the fragmentation and sub! speciation of their populations[ During the last glacial period of Pleistocene "07Ð19999 years ago# a large refugium of decidu! ous oak forests was present in western Iberia\ as a consequence of the warm Gulf Stream "Zagwijn 0881#[ After the last glacial period "Wurm# both P[ b[ boca`ei and P[ b[ carbonelli could expand from their refugia[ In accordance with the genetic dis! tance found\ the hypothesis of di}erent glacial refugia for each

6 099 SAł!SOUSA\ ALMEIDA\ ROSA\ VICENTE and CRESPO Table 5[ Nei s "0867# genetic matrices for four samples of Podarcis boca`ei] genetic distance above diagonal and genetic identity below diagonal Subspecies Sample G B P C P[ b[ boca`ei Gere¼s Ð 9[134 9[149 9[047 P[ b[ berlen`ensis Berlenga 9[672 Ð 9[900 9[000 P[ b[ berlen`ensis Peniche 9[668 9[878 Ð 9[979 P[ b[ carbonelli Cabo Raso 9[743 9[784 9[812 Ð lizard form might explain why so many islands o} north!west Galicia were exclusively colonized by P[ b[ boca`ei instead\ for instance\ by P[ hispanicus which is also found in the adjacent mainland nowadays[ Conversely\ south!west Galician islands were only colonized by P[ hispanicus "north!west Iberian form#\ to the detriment of P[ b[ boca`ei or P[ b[ carbonelli "see data from Galan 0874\ 0875\ 0876^ Mateo 0889\ 0886^ Perez!Mellado 0886^ Sa!Sousa 0884\ 0887\ 1999#[ The chance of one island being colonized by non~ying vertebrates might have been dependent on the abundance of the species that occurred in the adjacent mainland synchronically and with the formation of land!bridges "Cox and Moore 0882#[ Thus\ P[ b[ boca`ei may occur in the north!west Galician islands now for the simple reason that it colonized them _rst from its nearby refugium\ at the same time as P[ hispanicus started to colonize south!west Galicia "see Bas 0873^ Galan 0875#[ Probably\ P[ b[ carbonelli had a Wurmian refugium some! where further south\ towards the river Douro "see Fig[ 0#\ a hypothesis that might explain its genetic di}erentiation from P[ b[ boca`ei\ and also their parapatric distributions at present[ The genetic and morphological resemblance found between and within P[ b[ berlen`ensis:p[ b[ carbonelli populations seems compatible with the assumption of a recent postglacial separ! ation\ whereas other insular Podarcis populations were sep! arated by eustatic rising of sea level "see Guillaume and Lanza 0871^ Cirer and Guillaume 0875^ Ramon et al[ 0880^ Capula 0883#[ There are a few islands o} the west coast of Portugal on which only P[ b[ berlen`ensis "e[g[ Berlengas islands# or P[ b[ carbonelli "e[g[ Baleal\ near Peniche^ and Pessegueiro o} the south!west coast# are present\ although nowadays P[ hispanicus "south!west Iberian form# is the wall lizard species that is more abundant on the adjacent mainland "Sa!Sousa 0881\ 0884\ 1999^ cf[ Fig[ 0#[ Both postglacial warming and the progressive aridity in central and southern Iberia con_ned the areas in~uenced by the Atlantic during the Quaternary "Zagwijn 0881#[ This phenomenon should have led to a contraction of the range of P[ b[ carbonelli towards areas where Atlantic conditions still predominated[ However\ the known distribution of P[ b[ car! bonelli shown in Fig[ 0 is still patchy\ e[g[ in Beira Litoral\ Sistema Central range and other mountain areas "Sa!Sousa 0884\ 0888#\ as a consequence of the discontinuity of environ! mental conditions required by this lizard "e[g[ Estremadura and south!western coastlands# and the real di.culties encountered when trying to _nd them in the _eld[ In general\ we found some parallelism between P[ boca`ei and Schreiber s green lizard "Lacerta schreiberi# with respect to distribution and climatic requirements[ This latter species is also an IberianÐAtlantic endemism whose area has contracted but which still occurs in some isolated locations in south!west Iberia "De la Riva 0876^ Brito et al[ 0885^ Brito et al[ 0887#[ Finally\ taking into account the genetic di}erentiation between P[ b[ boca`ei and P[ b[ berlen`ensis:p[ b[ carbonelli\ the morphological resemblance of the two latter semi!species or the allopatric distribution "and presumed glacial refugia#\ we may hypothesize a species rank for P[ carbonelli\ with the Berlenga wall lizard as its insular subspecies\ P[ c[ berlen`ensis[ The comparatively lower genetic polymorphism "0[4 to 1 times# that we found for P[ b[ berlen`ensis could be the result of the founder e}ect[ The hypothesis proposed above seems a little speculative using only our present data\ but it is clearly supported by evidence from mitochondrial DNA analyses on these lizards conducted by James D[ Harris "personal communication#[ Acknowledgements We thank all colleagues and technicians who helped us in various ways\ either in the _eld or in the laboratory[ Lizards were captured and handled under permit from the Instituto de Conservacža½o da Natureza "ICN#[ P[ Sa!Sousa was supported by a grant of Fundacža½o para Cie¼ncia e Tecnologia "FCT# Praxis XXI!BD:4157:84\ while L[ Vicente and E[ G[ Crespo were supported by "FCT#PBIC:C:BIA:1984:84[ Zusammenfassung Genetische und morpholo`ische Verwandtschaftsbeziehun`en bei der Berlen`a Mauer!Eidechse "Podarcis boca`ei berlen`ensis] Lacertidae# Sechs biometrische Merkmale und 04 Allozym!Loci wurden bei den drei bekannten Unterarten von Podarcis boca`ei untersucht[ Im Gegen! satz zu den biometrischen Ergebnissen\ erweist sich P[ b[ berlen`ensis genetisch naher verwandt mi P[ b[ carbonelli "Nei s D 9[963Ð9[095# als mit der nominellen Unterart P[ b[ boca`ei[ Die biometrischen Ergeb! nisse sind wahrscheinlich durch den relativen Gro)wuchs von P[ b[ berlen`ensis verzerrt[ Die postglaziale Besiedlung von zwei verschiede! nen Refugien konnte vielleicht die genetische Di}erenzierung zwischen P[ b[ boca`ei und P[ b[ carbonelli "Nei s D 9[047# erklaren[ Sie konnte auch die bekannte Kolonisation dieser Formen uber kustennahe Inseln im Westen der Iberischen Halbinsel und deren gegenwartige Verbreitun! gen im Norden "Galizien\ N Portugal# bzw[ Im Suden "Zentral!Portu! gal\ W Sistema Central# des Flu)tals des Douro verstandlich machen[ References Almeida\ A[ P[^ Crespo\ E[ G[^ Vicente\ L[ A[\ 0881] Genetic study of Podarcis populations at Berlengas Archipelago[ In] Valakos\ E[ "ed[#\ Abstracts of First International Congress on the Lacertids of the Mediterranean Basin\ Mytilini\ Greece[ 02Ð06 April[ Mytilini] Soc[ Hellenic Zool[\ p[ 1[ Bas\ S[ L[\ 0873] Biogeograf(a de los an_bios y reptiles de Galicia\ un ensayo de sintesis[ Amphibia!Reptilia 4\ 178Ð509[ Bobyn\ M[ L[^ Darevsky\ I[ S[^ Kupriyanova\ L[ A[^ MacCulloch\ R[ D[^ Upton\ D[ E[^ Danielyan\ F[ D[^ Murphy\ R[ W[\ 0885] Allozyme variation in populations of Lacerta raddei and Lacerta nairensis "Sauria] Lacertidae# from Armenia[ Amphibia!Reptilia 06\ 122Ð135[ Brito\ J[ C[^ Brito e Abreu\ F[^ Paulo\ 9[S[^ Rosa\ H[ D[^ Crespo\ E[ G[\ 0885] Distribution of Schreiber s Green lizard "Lacerta schreiberi# in Portugal] a predictive model[ Herpetol[ J[ 5\ 32Ð36[ Brito\ J[ C[^ Paulo\ O[ S[^ Crespo\ E[ G[\ 0887] Distribution and habitats of Schreiber s Green lizard "Lacerta schreiberi# in Portugal[ Herpetol[ J[ 7\ 076Ð083[ Busack\ S[ D[\ 0875] Biogeographic analysis of the herpetofauna sep! arated by the formation of the Strait of Gibraltar[ Natl[ Geographic Res[ 1\ 06Ð25[ Busack\ S[ D[\ 0876] Morphological and biochemical di}erentiation in Spanish and Moroccan populations of the lizard\ Lacerta lepida[ J[ Herp[ 10\ 166Ð173[

7 Genetic and morphological relationships of Berlenga wall lizard 090 Capula\ M[\ 0883] Genetic variation and di}erentiation in the lizard\ Podarcis wa`leriana "Reptilia] Lacertidae#[ Biol[ J[ Linn[ Soc[ 41\ 066Ð085[ Case\ T[ J[^ Schwaner\ T[ D[\ 0882] Island:mainland body size di}er! ences in Australian varanid lizards[ Oecologia 83\ 091Ð098[ Cheylan\ M[\ 0877] Variabilite phenotipique du lezard des murailles Podarcis muralis sur les (¼les de la co¼te provencžale[ Rev[ Ecol[ "Terre Vie#\ Paris 32\ 176Ð210[ Cirer\ A[ M[\ 0870] La Lagartija Ibicenca y su C(rculo de Razas[ Ibiza] Consell Insular d Eivissa y Formentera[ Cirer\ A[ M[\ 0886] Podarcis pityusensis "Bosca\ 0772#[ Lagartija de las Pitiusas\ lagartixa das Pitiusas[ In] Pleguezuelos\ J[ M[ "ed[#\ Distribucion Y Biogeograf(a de Los An_bios Y Reptiles En Espan½a Y Portugal[ Granada] University of Granada and Asoc[ Herp[ Esp[ "AHE#[ pp[ 253Ð255[ Cirer\ A[ M[^ Guillaume\ Cl[!P[\ 0875] Electrophoretic analysis of the Pityusic islands lizard[ In] Rocek\ Z[ "ed[#\ Studies in Herpetology[ Prague] Charles University\ pp[ 190Ð195[ Cirer\ A[ M[^ Mart(nez!Rica\ J[ P[\ 0889] The polymorphism of Podarcis pityusensis and its adaptative evolution in Mediterranean Isles[ Her! petol[ J[ 0\ 354Ð362[ Cox\ C[ B[^ Moore\ P[ D[\ 0882] Biogeography Ð an Ecological and Evolutionary Approach[ Oxford] Blackwell Scienti_c Publications[ De la Riva\ I[\ 0876] Zoogeogra_a de Lacerta schreiberi Bedriaga\ 0767[ Rev[ Esp[ Herp[ 1\ 38Ð58[ Delauguerre\ M[^ Cheylan\ M[\ 0881] Atlas de Repartition des Batra! ciens et Reptiles de Corse[ Ajaccio "Corsica#] Parc Nat[ Reg[ Corse and E cole Pratique des Hautes E tudes "Montpellier#[ Fu\ J[^ Darevsky\ I[ S[^ MacCulloch\ R[ D[^ Kupriyanova\ L[ A[^ Roit! berg\ E[ S[^ Sokolova\ T[ M[^ Murphy\ R[ W[\ 0884] Genetic and morphological di}erentiation among Caucasian rock lizards of the Lacerta caucasica complex[ Russ[ J[ Herp[ 1\ 25Ð31[ Galan\ P[\ 0874] Morfologia de las Podarcis boca`ei "Seoane\ 0773# "Sauria] Lacertidae# de dos islas atlanticas ibericas] Sisargas y Berlen! gas[ Trabajos Compostelanos Biologia 01\ 010Ð028[ Galan\ P[\ 0875] Morfologia y distribucion del genero Podarcis\ Wagler\ 0729 "Sauria] Lacertidae# en el noroeste de la Pen(nsula Iberica[ Rev[ Esp[ Herp[ 8\ 46Ð64[ Galan\ P[\ 0876] Notas preliminares sobre la herpetofauna de las islas C(es y Ons[ Mustela 2\ 53Ð58[ Galan\ P[\ 0886] Podarcis boca`ei "Seoane\ 0773#[ Lagartija de Bocage\ lagartixa de Bocage[ In] Pleguezuelos\ J[ M[ "ed[#\ Distribucion Y Biog! eograf(a de Los An_bios Y Reptiles En Espan½a Y Portugal[ Granada] University of Granada and Asoc[ Herp[ Esp[ "AHE#[ pp[ 126Ð 131[ Gorman\ G[ C[^ Soule\ M[^ Yang\ S[ Y[^ Nevo\ E[\ 0864] Evolutionary genetics of insular Adriatic lizards[ Evolution 18\ 41Ð60[ Guillaume\ Cl[!P[\ 0878] Utilisation de quelques techniques recentes non morphologiques en systematique et phylogenie des amphibiens et des reptiles] quelques exemples "1eme partie#[ Bull[ Soc[ Herp[ Fr[ 49\ 08Ð31[ Guillaume\ Cl[!P[^ Cirer\ A[ M[\ 0874] Comparacion electroforetica de diez poblaciones de Podarcis pityusensis Bosca\ 0771 "Lacertidae# de Ibiza\ Formentera e islotes circunvecinos[ Bull[ Inst[ Cat[ Hist[ Nat[ 41 "Sec[ Zool[# 5\ 086Ð197[ Guillaume\ Cl[!P[^ Lanza\ B[\ 0871] Comparaison electrophoretique de quelques especes de lacertides mediterraneens\ genera Podarcis et {Archaelacerta [ Amphibia!Reptilia 3\ 250Ð264[ Guillaume\ Cl[!P[^ Pasteur\ N[^ Bons\ J[\ 0865] Distinction par elec! trophorese sur gel d amidon des especes de lezards Lacerta muralis Laurenti 0657 et Lacerta hispanica Steindachner 0769 dans les popu! lations sympatriques d Espagne et du Languedoc!Roussillon[ C[ R[ Acad[ Sc[ Paris] D 171\ 174Ð177[ Legendre\ P[^ Legendre\ L[\ 0887] Numerical Ecology[ Amsterdam[ Elsevier Science B[V[ MacCulloch\ R[ D[^ Fu\ J[^ Darevsky\ I[ S[^ Danielyan\ F[ D[^ Murphy\ R[ W[\ 0884] Allozyme variation in three closely related species of Caucasian rock lizards "Lacerta#[ Amphibia!Reptilia 05\ 220Ð 239[ Magraner\ J[\ 0875] Nouvelle donnee sur la repartition de Podarcis boca`ei carbonelli "V[ Perez!Mellado\ 0870#\ "Sauria\ Lacertidae#\ dans la Pen(nsule Iberique et observations sur son ecologie a Don½ana "Andalousie\ Espagne#[ Bull[ Soc[ Herp[ Fr[ 27\ 5Ð01[ Mateo\ J[ A[\ 0889] Aspectos biogeoga_cos de la fauna reptiliana en las islas espan½olas[ Rev[ Esp[ Herp[ 3\ 22Ð33[ Mateo\ J[ A[\ 0886] Las islas e islotes del litoral iberico[ In] Pleguezuelos\ J[ M[ "ed[#\ Distribucion Y Biogeograf(a de Los An_bios Y Reptiles En Espan½a Y Portugal\ Granada] University of Granada and Asoc[ Herp[ Esp[ "AHE#[ pp[ 232Ð249[ Mayer\ W[^ Arribas\ O[ J[\ 0885] Allozyme di}erentiation and relation! ship among the Iberian!Pyrenean mountain lizards "Squamata] Sau! ria] Lacertidae#[ Herpetozoa 8\ 46Ð504[ Mayer\ W[^ Tiedemann\ F[\ 0871] Chemotaxonomical investigations in the collective genus Lacerta "Lacertidae] Sauria# by means of protein electrophoresis[ Amphibia!Reptilia 1\ 238Ð244[ Nei\ M[\ 0867] Estimation of average heterozigosity and genetic distance from a smaller number of individuals[ Genetics 78\ 472Ð489[ Pasteur\ N[^ Pasteur\ G[^ Bonhomme\ F[^ Catalan\ J[^ Britton!Davidian\ J[\ 0876] Manual Technique de Genetique par E lectrophorese des Proteines[ Tec[ + Doc[ Paris] Lavoisier[ Perez!Mellado\ V[\ 0870a] La lagartija de Bocage\ Podarcis boca`ei "Seoane\ 0773#[ Primeros datos sobre su distribuc(on\ colorido y ecologia[ Amphibia!Reptilia 0\ 142Ð157[ Perez!Mellado\ V[\ 0870b] Nuevos datos sobre la sistematica y dis! tribuc(on de Podarcis boca`ei "Seoane\ 0773# "Sauria] Lacertidae# en la Pen(nsula Iberica[ Amphibia!Reptilia 1\ 148Ð154[ Perez!Mellado\ V[\ 0872] La herpetofauna de Salamanca] un analisis biogeogra_co y ecologico[ Rev[ Provincial de Estudios\ Salamanca 8:09\ 8Ð67[ Perez!Mellado\ V[\ 0873] Sobre los anf(bios y reptiles de la Sierra de la Estrela "Beira Alta\ Portugal#[ Bull[ Soc[ Cat[ Ictio[ Herp[ 7\ 02Ð19[ Perez!Mellado\ V[\ 0875] Podarcis boca`ei "Seoane 0773# Nordwes! tiberiche Mauereidechse[ In] Bohme\ W[ "ed[#\ Handbuch der Repti! lien und Amphibien Europas\ 1[ Wiesbaden] Aula!Verlag[ pp[ 04Ð13[ Perez!Mellado\ V[\ 0886] Podarcis boca`ei "Seoane\ 0773#[ In] Gasc\ J[! P[^ Cabela\ A[^ Crnobrnja!Isailovic\ J[^ Dolmen\ D[^ Grossenbacher\ K[^ Ha}ner\ P[^ Lescure\ J[^ Martens\ H[^ Mart(nez!Rica\ J[ P[^ Maurin\ H[^ Oliveira\ M[ E[^ So_anidou\ T[ S[^ Veith\ M[^ Zuiderwijk\ A[\ "eds#\ Atlas of Amphibians and Reptiles in Europe[ Paris] Mus[ Nat[ Hist[ Nat[ Paris and Societas Europaea Herpetologica "SEH#[ pp[ 161Ð162[ Perez!Mellado\ V[^ Galindo\ M[ P[\ 0875] Sistematica de Podarcis "Sau! ria] Lacertidae# Ibericas y Norteafricanas mediante Tecnicas Multi! dimensionales[ Ser[ Manuales Universitarios[ Salamanca] University of Salamanca "Spain#[ Perez!Mellado\ V[^ Salvador\ A[\ 0877] The Balearic lizard Podarcis lilfordi "Gunther\ 0763# "Sauria] Lacertidae# of Menorca[ Arq[ Mus[ Bocage n[s[ 0\ 016Ð084[ Ramon\ M[ M[^ Castro\ J[ A[^ Arranz\ M[ J[\ 0880] A study of the Balearic lizard Podarcis lilfordi based on morphological characters and enzyme polymorphisms[ Evolucion Biologica 4\ 64Ð81[ Rosa\ H[^ Oliveira\ M[ E[\ 0883] Genetic di}erentiation of the Iberian tree frogs Hyla arborea molleri and Hyla meridionalis "Amphibia] Anura#[ Z[ Zool[ Syst[ Evolut[!Forsch[ 21\ 006Ð017[ Sa!Sousa\ P[\ 0881] The populations of lizards "Podarcis# at Peniche[ In] Valakos\ E[ "ed[#\ Abstracts of First International Congress on the Lacertids of the Mediterranean Basin\ Mytilini\ Greece[ 02Ð06 April[ Mytilini] Soc[ Hellenic Zool[ p[ 17[ Sa!Sousa\ P[\ 0884] Coastland distribution of the Bocage mountain lizard\ Podarcis boca`ei carbonelli in Portugal[ In] Vicente\ L[ A[ "ed[#\ Abstracts of Second International Congress on the Lacertids of the Mediterranean Basin\ Olha½o\ Portugal[ 14Ð29 April[ Olha½o] P[ N[ Ria Formosa\ p[ 24[ Sa!Sousa\ P[\ 0887] Distribucion de la lagartija Podarcis b[ boca`ei en Portugal[ Bol[ Asoc[ Herp[ Esp[ 8\ 1Ð3[ Sa!Sousa\ P[\ 0888] New data on the distribution of Podarcis boca`ei carbonelli Perez!Mellado\ 0870 in Portugal[ Herpetozoa[ 01\ 76Ð89[ Sa!Sousa\ P[\ 1999] A predictive distribution model for the Iberian wall lizard "Podarcis hispanicus# in Portugal[ Herpetol[ J[ 09\ 0Ð00[ Salvador\ A[\ 0875] Podarcis lilfordi "Gunther\ 0763# Balearen!Eidechse[ In] Bohme\ W[ "ed[#\ Handbuch der Reptilien und Amphibien Euro! pas\ 1[ Wiesbaden] Aula!Verlag[ pp[ 72Ð009[ Thorpe\ R[ S[\ 0874] Body size\ island size and variability in the Canary Island lizard of the genus Gallotia[ Bonn[ Zool[ Beitr[ 25\ 370Ð376[ Vicente\ L[ A[\ 0874] Description d une nouvelle sous!espece de Podarcis

8 091 SAł!SOUSA\ ALMEIDA\ ROSA\ VICENTE and CRESPO boca`ei "Seoane\ 0773# "Sauria] Lacertidae# de l (¼le de Berlenga] Pod! arcis boca`ei berlen`ensis[ Bull[ Mus[ natn[ Hist[ nat[\ Paris] 3e ser[ 6 "section A#\ 156Ð163[ Vicente\ L[ A[\ 0878] Sobre a Historia Natural dos Repteis da Ilha Berlenga] a s(ndrome de insularidade[ Doctoral Thesis\ Lisbon\ Por! tugal] FCUL\ University of Lisbon[ Zagwijn\ W[ H[\ 0881] Migration of vegetation during the Quaternary in Europe[ Cour[ Forsch[ Senck[ 042\ 8Ð19[ Authors addresses] P[ Sa!Sousa "for correspondence#\ Centro de Eco! logia Aplicada\ Universidade de E vora\ Largo dos Colegiais 1\ P!6991Ð 433 E vora\ Portugal[ E!mail] psasousaýyahoo[com^ A[ P[ Almeida\ H[ D[ Rosa\ L[ Vicente and E[ G[ Crespo\ Centro de Biologia Ambiental\ Faculdade de Cie¼ncias da Universidade de Lisboa\ bloco C1 piso 2>\ Rua Ernesto de Vasconcelos\ Campo Grande\ P!0638Ð905 Lisboa\ Portugal Appendix 0 Descriptive statistics for six biometric characters measured in four samples of Podarcis bocagei "m\ males^ f\ females#[ Biometric characters are described in the text Podarcis b[ boca`ei Podarcis b[ berlen`ensis Podarcis b[ carbonelli Gere¼s Berlenga Peniche Cabo Raso G B P C n f m Variables x SD x SD x SD x SD SVL f 44[01 2[33 45[14 3[28 45[06 1[33 49[27 1[91 m 59[96 0[87 50[95 1[6 43[91 1[88 40[70 1[3 HL f 00[11 9[43 00[37 9[33 00[1 9[30 09[57 9[23 m 02[87 9[52 03[35 9[53 01[85 9[50 01[44 9[44 HW f 4[42 9[30 4[53 9[17 4[34 9[13 4[05 9[12 m 5[60 9[22 6[1 9[34 5[15 9[32 5[08 9[24 HD f 4[97 9[18 4[26 9[2 4[08 9[10 3[7 9[11 m 5[01 9[26 5[67 9[35 5[03 9[34 5[94 9[3 FLL f 05[17 9[72 03[82 0[48 04[13 9[5 02[56 9[47 m 08[26 9[81 08[03 9[73 06[34 0[21 05[03 9[57 HLL f 13[32 0[08 12[47 0[60 12[54 0[15 19[1 9[71 m 29[83 0[48 29[94 0[51 16[00 0[55 13[80 0[01 Appendix 1 Protein systems studied in the electrophoresis and respective loci and buffer systems Protein system E[C[ number Loci scored Organic tissues Bu}er system Adenylate kinase 1[6[3[2 AK L I Aspartate aminotransferase 1[5[0[0 AAT!0\1 H\ M I Creatine kinase 1[6[2[1 CK L I Esterases 2[0[0[0:1 EST!1 L II Lactate dehydrogenase 0[0[0[16 LDH!0\1 H\ M I Leucyl!tyrosine peptidade 2[3[0[0 PEP L III Malate dehydrogenase 0[0[0[26 MDH!0 L II Mannose phosphate isomerase 4[2[0[7 MPI L III Phosphogluco!mutase 1[6[4[0 PGM!1 L I Phosphogluconate dehydrogenase 0[0[0[32 PGD L I Superoxide dismutase 0[04[0[0 SOD L I Albumin Ð ALB B IV Hemoglobin Ð HB B IV B blood^ H heart^ L liver^ M muscle[ I tris!citrate ph 7[9^ II tris!citrate ph 5[6^ III tris!borate!lioh ph 7[2^ IV tris!barbital ph 7[7[