Contributions to Zoology, 65 (1) 5-34 (1995) SPB Academic Publishing bv, Amsterdam Morphological and allozyme studies of midwife toads (genus Alytes), including the description of two new taxa from Spain J.W. Arntzen & M. García-París 1 School of Biological Sciences, University of Wales, Bangor LL572UW, United Kingdom; 2 Museo Nacional de Ciencias Naturales, C.S.I.C., José Gutiérrez Abascal 2, 28006 Madrid, Spain Keywords: Allozymes, Alytes, midwife toads, morphometry, Spain, taxonomy Abstract cies A. o. obstetricans y A. o. boscai Lataste, 1879, descritas previamente, más una nueva subespecie del Noreste Ibérico descrita aquí como A. o. almogavariin. ssp. El estudio de la Allozyme variation in 31 to 50 presumptive loci of 12 popula- variación morfológicaentre especies y subespecies realizado me- tions of European midwife toads ofthe genus Alytes show ap- diante análisis de componentes principales y análisis discrimi- preciable genetic divergences ( from 0.29 to 0.72) among DNei de manifiesto la existencia de destacada simi- nante pone una four groups. These groups correspond to A. cisternasii Boscá, litud morfológicaentreA. obstetricans y A. dickhilleni que, por 1879, A. obstetricans (Laurenti, 1768), A. muletensis (Sanchiz lo tanto, pueden considerarse como especies crípticas desde el & Adrover, 1979), and a new species from the Betic mountains punto de vista morfológico. El análisis filogenético realizado described here asA. dickhilleni n. sp. Smaller divergences among datos usando A . cisternasii con electroforéticos, como grupo ex- geographic ofpopulations of A. obstetricans (up to groups DNei terno, confirma la existencia de estrechas relaciones filogenéti- = 0.17) support the existence of three geographic units in cas entre A. dickhilleni y A. muletensis queconstituyenun grupo Europe corresponding to the previously recognized subspecies A. monofilético, mientras que obstetricans se sitúa como grupo A. o. obstetricans and A. o. boscai Lataste, 1879, plus a third hermano del A. dickhilleni-A. muletensis. Adicionalmente par one described here under the designation ofA. o. almogavarii se discute la biogeografía histórica del género. The of variation n. ssp. analysis morphological among taxa discriminant shows using principal component and analysis a remarkable similarity between A. dickhilleni and A. obstetri- A these are cryptic species. phylogenetic analysis of allo- cans; Introduction zyme data using A. cisternasii as the outgroup supports a sister taxon relationship between A. muletensis and A. dickhilleni, with A. obstetricans the sister taxon to this clade. The historical A population of midwife toads was recently discov- biogeography of the is discussed. of the Balearic Islands genus ered on Mallorca, the largest (Alcover & Mayol, 1980; Mayol & Alcover, 1981). Surprisingly, these midwife toads appeared to be Resumen the survivors of the Mallorcan Baleaphryne mule- tensis described as fossil species by Sanchiz & El análisis de la variación genética en 12 poblaciones de sapos Adrover (1979). The taxonomie status of the parteros europeos del género Alytes, basado en el estudio de Mallorcan midwife toad as Alytes muletensis is now 50 31 a loci mediante electroforesis de proteínas, muestra la ex- generally accepted (García-París, 1985; Salvador, istencia de una divergencia genética = 0.29— pronunciada (DNei Two continental of midwifetoads de 1985). species are 0.72) entre cuatro grupos poblaciones. Estos grupos cor- the Iberian midwife toad respondena las especies Alytes cisternasii Boscá, 1879, A. obste- currently recognized, tricans (Laurenti, 1768), A. muletensis (Sanchíz & Adrover, A. cisternasii Boscá, 1879, from the southwestern 1979) y una especie nueva que se describe en este trabajo bajo region of the Iberian Peninsula, and the Common el nombre de A. dickhilleni n. sp. Las diferencias genéticas ob- midwife toad A. obstetricans (Laurenti, 1768) with servadas de entre grupos poblaciones geográficamente separa- a wide distribution that from the Middle das dentro de la A . obstetricans = ranges especie (hasta DNei 0.17) per- Atlas in Morocco the Harz Mountains in Ger- mite reconocerotros tres grupos correspondientes a las subespe- to 6 J. W. Arntzen & M. Garcia-Paris - Midwife toad systematics linewith its distribution, A. obstetri- many. In wide cans occupies a wide variety of habitats, ranging from rocky areas near mountain lakes, humid forests along coastal streams, to arid plateaus and, occasionally, xerophilous coastal scrub zones close to man-made ponds. In the southernmost part of the Iberian Peninsula, midwife toads appear to be restricted to the higher altitudesof mountainranges (Antunez et al., 1982), where the toads may live under ecological conditions that are surprisingly similarto thoseof A. muletensison Mallorca. Mor- phological and allozyme variation has been report- ed for A. obstetricans (Lataste, 1879a; Pasteur & Bons, 1962; Guyétant, 1977; Arntzen & Szymura, 1984; Viegas & Crespo, 1985; Rosa et al., 1990) and is reflected in the description of several subspecies. The known distributions of revised midwife toad species and subspecies are presented in Fig. 1. Immunological, protein electrophoretic and mor- A. dickhilleni. - Spain: A, Sierra de Alcaraz (present paper); B, Sierra de Gádor (Antúnez et al., 1988); C, Sierra Tejeda (present paper). A . muletensis. — In its present-day distribution the species is only known from six localities in the Serra Tramuntana of Mal- lorca, Spain. A. - obstetricans. France: 1, map of the “Institut Géogra- phique National” (IGN) 1/50,000 XXII-2 (Calais) (Guyétant, 1989). The Netherlands: 2, Limburg, gridreference 60.42 (Berg- mans & Zuiderwijk, 1986). Germany: 3, Westfalia, MTB grid reference 3819/4(Feldmann, 1981);4, Niedersachsen, MTB grid Fig. 1. Distribution of four species of midwife toads. Reliable 3723/2 (Lemmel, 1977); 5, MTB grid 4132/2 (Schiemenz & records the of the documenting outer range species are as Günther, 1994); 6, MTG grid 4434/1 (Schiemenz & Günther, follows: 1994); 7, MTB grid 5330/4 (Schiemenz & Günther, 1994); Alytes cisternasii. - Portugal: transect a-a’ according to Rosa 8, UTM grid QA 04 (Müller, 1976);9, UTM grid PV 54 (Müller, et al. (1990). Spain: b, Province of Zamora: UTM grid TL 69 1976). Switzerland: 10, Berneck (Grossenbacher, 1988); (Pollo et al., 1988); c, Province of Valladolid: UTM grid UL 46 11, Grindelwald (Grossenbacher, 1988); 12, Vevey (Grossen- et (García-París al., 1990); d, Province of Segovia: UTM grid bacher, 1988). France: 13, map IGN XXXIV-31 (Annecy-Ugine) UL 71 et Avila (García-París al., 1990); e, (Arntzen, 1979); (Guyétant, 1989); 14, map IGN XXXV-45 (St. Tropez) f, Province of Avila: UTM grid TK 88 (García-París et al., (Guyétant, 1989). Spain: 15, Province of Alicante: Alcoy 1990); g, Province ofToledo: UTM grid UK 03 (García-París et (García-París et al., 1993); 16, Province of Cuenca: Uña al., 1990); h, Province of Madrid: UTM grid UK 99 (García- (García-París, unpublished data); 17, Province of Guadelajara: et París al., 1989); i, Province of Madrid: UTM grid VL 44 Ciruelos (J. Prieto & G. Astudillo, pers. comm., 1990); (García-París et al., 1989); j, Province of Guadalajara: UTM 18, Province of Madrid: UTM grid UK 99 (García-París et al., grid VL 81 (García-París et al., 1990); k, UTM grid VH 75 1989); 19, river Tormes, Sierra de Gredos (Arntzen, 1979). Por- (Pleguezuelos & Moreno, 1990); 1, Province of Toledo: UTM tugal: transect 20-20’ according to Rosa et al. (1990); 21, isolat- UK 42 et Province of Madrid: grid (García-París al., 1990); m, ed occurrence on the Serra de S. Mamede (Rosa et al., 1990). VK & UTM grid 73 (García-París et al., 1990); n, Toledo (Boscá, Morocco: 22, Tleta Ketama (Pasteur & Bons, 1962; Arntzen 1880b); of Jaén: UTM grid o, Province VH 75 (Pleguezuelos & Szymura, 1984; cf. Bons, 1973);23, Merhraoua (Libis, 1985; cf. Moreno, 1990); p, Province of Córdoba: UTM grid UH 40 Bons, 1973); 24, Djebel Tazekka, Taza (Mellado& Mateo, 1992; 1980); Province of Huelva: El Rocío cf. Chefchaouèn (López-Jurado et al., q, Bons, 1973); 25, (García-París, pers. obs., (Collado et al., 1976). 1993; cf. Bons, 1973). - 7 Contributions to Zoology, 65 (I) 1995 phological studies identify A. muletensis and A. Alytes obstetricans: France: Jublains, N = 5 (e, m) ZMA 7642. 8156, N = 5 (m) ZMA 7584, N = 2 (m) ZMA Germany: obstetricans as sister taxa (Clarke, 1984; Crespo et Asturias: Picos Köln, N = 4 (m) MNCN uncatalogued. Spain: al., 1984; Hemmer, 1984; Maxson, 1984; Viertel, de Europa, N = 1 (e) ZMA 8157; Asturias and León: Puerto de 1984). On the other hand, cytogenetic data suggest = San Isidro, N = 7 (e, m) MNCN uncatalogued,N 10 (m); that cisternasii and A. obstetricans are a sister A. N = Navarra: lturen, N = 11 (m) MNCN uncatalogued; Irati, while = 5 MNCN group (Herrero, 1984), some morphological 5 (e, m) MNCN 16778 and uncatalogued,N (m) un- = 16774-79, data ambiguously point to either A. cisternasii or catalogued; Barcelona: Berga, N 5 (e, m) MNCN = MNCN N = 1 (e, m) MNCN 16743-45 and 16760-63, N 3 (m) A. muletensis as the sister taxon of A. obstetricans N = 4 16746-48, N = 10 (m); Barcelona, Universidad Central, exists for (Sanchiz, 1984). No support grouping (m) MNCN 16749-52; Pontevedra: Tuy, N = 6 (e, m) MNCN together A. muletensisand A. cisternasii. 16780 and uncatalogued; Bayona, N = 9 (m); Castellón: to elucidate In the work we (1) = attempt = present Caudiel, N 2(e, m) MNCN uncatalogued; Benicassim, N the phylogenetic relationships of the species in the 2 (e, m) MNCN 16767 and 16773, N = 26 (m). describe variation in Midwife toads from the Betic mountains, Spain, are from: genus Alytes, (2) geographical Albacete: Bienservida, N = 8 (e, m) MNCN 16768-72, MNCN A. obstetricans, the most variable of midwife toad N 1 = 4 16739-41, = (m) MNCN 16742, N (m); Paterna del species, and (3) find readily applicable features in Madera, N = 2 (m) MNCN 16781-82, N = 17 (m); Málaga: external morphology that allow correct identifica- Alcaucín, N = 4 (m) MNCN 16735-38; Canillas de Aceituno, of and As the result of the tion species subspecies.