Bijdragen tot de Dierkunde, 63 (1) 3-14 (1993)

SPB Academie Publishing bv, The Hague

Morphological characterization, cytogenetic analysis, and geographical

distribution of Marbled Newt Triturus the Pygmy marmoratus pygmaeus

(Wolterstorff, 1905) (Caudata: Salamandridae)

M. García-París P. Herrero C. Martín J. Dorda M. Esteban & B. Arano

Museo Nacional de Ciencias Naturales, José Gutiérrez Abascal 2, E-28006 ,

Keywords: Taxonomy, cytogenetics, Salamandridae, Triturus,

T. m. mar- T. Abstract moratus o a m. pygmaeus. Estos rasgos se aplican a series de

individuos procedentes de colecciones científicas o bien observa-

dos directamente sobre el terreno. Como consecuencia de la Triturus marmoratuspygmaeus, a problematicsubspecies of the

aplicación de estos criterios, el área de distribución de T. m. pyg- Marbled Newt from the southern part of the Iberian Peninsula,

maeus se extiende considerablemente hacia el norte. La distribu- is redescribed using specimens collected in the “typical” area. la ción de T. m. marmoratus incluye mitad septentrional de la external features Diagnostic morphological are provided to per- Península Ibérica y la mayor parte de Francia, mientras que T. mit the accurate determination ofthe specimens belonging either

m. pygmaeus ocupa unaampliaporción en la región sudocciden- T. T. These to m. marmoratus or to m. pygmaeus. diagnostic

tal de la Península Ibérica. La zona de contacto entre ambas features were applied to individuals both from the field and

subespecies parece localizarse a lo largo del en from museum collections. The results indicate a larger distribu-

Portugal y España. T. m. marmoratus sobrepasa hacia el sur el tional area for T. m. pygmaeus than was previously recognized. Sistema Central en tres áreas: (), Sierra The distribution of T. m. marmoratus ranges over the northern de Gata El único (España) y (España). half of the Iberian Peninsula and most of France; T. m. pyg-

punto en el que T. m. pygmaeus sobrepasa hacia el norte estas maeusoccupies the southwestern part of the Iberian Peninsula. montañas, se sitúa en las proximidades del Puerto de Malagón The contact areabetween the two subspecies seemsto be located

(Madrid, España). No se han observado casos de simpatría es- alongthe Central Range Mountains (Sistema Central) in Portu- ni individuos tricta, con rasgos intermedios entre ambas gal and Spain. T. m. marmoratus extends southwards beyond

subespecies. Los resultados de un extenso análisis citogenético this borderline in three areas: Serra da Estrela (Portugal), Sierra

no muestran la existencia de diferencias entre T. m. marmoratus de Gata (Spain) and Sierra de Guadarrama (Spain). The only T. si bien y m. pygmaeus, la población de T. m. pygmaeus de point at which T. m. pygmaeus reaches northwards beyond the

Doñana (Huelva, España) muestra un patrón particular, aunque Central System is near Puerto de Malagón (Madrid Province, de bandas poco diferenciado, C. of strict individuals Spain). No cases sympatry, nor with inter-

mediate morphologicalfeatures have been observed. The results

of an extensive cytogenetical analysis do not show any dif- Introduction ferences between T. m. pygmaeus and T. m. marmoratus. In-

the T. from terestingly, however, m. pygmaeus populations

Doñana (Huelva Province, Spain) showed an exclusive, though The marbled newt Triturus marmoratus (Latreille, little differentiated, C-banding pattern. 1800), occurs over a large part of western Europe,

from northwestern and central France in the north

to southern Portugal and southwestern Spain in Resumen the south. Most authors accept the existence of

two subspecies, T. m. marmoratus, widely spread redescribe Se Triturus marmoratus pygmaeus, Una subespecie over most of the area described, and T. m. pyg- problemática de triton jaspeado del sur de la Península Ibérica, maeus (Wolterstorff, 1905), which is limited to the utilizando para ello especímenes procedentes de su área “típi- southern part of the Iberian Peninsula (Macgregor ca”. Se definen caracteres morfológicos externos que permiten la identificación de los individuos pertenecientes a et al., 1990).

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Table I. External of of T. from Cantabria and T. from Cádiz measurements (in mm) specimens m. marmoratus m. pygmaeus (Spain).

are: = = = Variables measured TL = total length; SV snout-vent length; FHL distance between forelimbs and hindlimbs; FLL fore-

HLL = 3HF HW HL limb length; 3FF = third finger length; hindlimb length; = third toe length; = head width; = head length.

TL SV FHL FLL 3 FF HLL 3HF HW HL

T. m. marmoratus

Males mean 132.1 72.9 37.5 29.3 8.8 28.5 9.5 12.8 15.6

n = 20 SD 9.04 6.16 2.90 2.56 1.2 2.86 1.0 1.20 1.72

Females mean 148.4 81.5 47.2 29.7 7.9 29.2 8.0 14.3 17.9

n = 15 SD 10.45 5.47 3.69 2.82 0.9 1.83 0.7 1.22 1.48

T. m. pygmaeus

Males mean 106.5 59.4 28.9 21.6 6.5 11.7 7.2 10.7 15.1

n = 15 SD 9.02 5.35 4.45 1.63 0.8 1.77 0.7 1.21 1.42

Females mean 116.5 61.1 33.7 21.0 5.8 20.9 6.0 11.8 15.9

n = 10 SD 5.01 3.17 2.15 1.28 0.8 1.35 0.4 0.85 1.58

The most recent studies on the morphology, im- Material and methods munology, and genetics of T. marmoratus and allied species (Dorda & Esteban, 1986; Busack et Twenty-five specimens, collected during their aquatic phase in

San and Los Barrios Cádiz al., 1988; Macgregor et al., 1990) point to the com- Roque (Algeciras Bay, Province,

used for the of Spain), were redescription T. m. pygmaeus. plexity of the group and suggest that a revision of

These capture sites were selected by consideration of two facts: the taxonomie status of T. m. pygmaeus may be ap- firstly, the "terra typicarestricta" for T. m. pygmaeus as report- propriate. ed by Mertens & Miiller (1928) is Cádiz, and within the Cádiz

One of the is the in problems difficulty correctly Province, Algeciras is one of the two sites mentioned by

Wolterstorff the external features of the in- identifying individuals belonging to each subspe- (1905). Secondly,

dividuals from these populations are similar to those indicated cies. This seems partly due to the lack of reliable by Wolterstorff in the original description. diagnostic features, as well as to an extended mis- Thirty-five T. m. marmoratus individuals from different sites interpretation of Wolterstorff's abbreviated (1905) in Cantabria (northern Spain) were used as specimens for com-

of m. description T. pygmaeus. This misinterpreta- parison in order to establish the diagnosticfeatures of T. m. pyg-

Some individuals of tion arises from the confusion of the extremely maeus. both groups were kept alive until the

end ofthe breedingperiod in order to describe their colouration small size of specimens of the coastal populations during the terrestrial phase. Measurements and description of from southwestern Spain with that of the small external features and colouration were all made on live speci- individuals originally described Wolterstorff by mens. External measurements are indicated in Table I. Statisti-

(1905). cal analysis is restricted here to the use ofthe /-test for compari-

son of mean values (Zar, 1984).

Once the diagnostic features were established, we proceeded In the present study the southern subspecies

to revise the material available from some museum collections is (T. m. pygmaeus) redescribed using specimens and to conduct a field inspection of selected populations(cf. Ap- from the "typical" area. Some diagnostic morpho- pendix). After the subspecific assignation, 255 specimens from logical and features of T. cytogenetic m. pygmaeus diverse localities were measured preserved or alive. A two-group

T. are proposed that allow its correct identification. comparison of the resultant T. m. marmoratus and m. pyg-

conducted discriminant anal- The maeus samples was using stepwise diagnostic features are applied to individuals from the Discriminant ofBMDP ysis program package (Dixon, collected in the field and from museum collec-

1975). Total length was excluded from this analysis. tions and the resultant distributional limits are in- For use in cytogenetic analysis, 80 males and 73 females from dicated. 22 localities were injected with 0.05 ml of a

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solution of 24 h before that of with an 0.1% colchicine per gram body weight, a light yellowish background orange they were sacrified. Testes and intestine were removed and hue which several and white on large blackish spots placed in 0.05 M KC1 for 5 minutes, subsequently fixed in a solu- dots may be found. These white dots usually show 48 tion of ethanol and acetic acid (3:1) for a minimum of h, and the flanks and a higher density on occasionally then stored in 70% ethanol at 4°C. Preparations of mitotic chro- the colouration. and cover background mosomes were made using the squash technique were air completely

White dots also the base of the dried after immersion in liquid nitrogen. The slides were are present on

C-banding method described by Sumner neck the processed followingthe flanks, spreading over the sides of the and

(1972) with slight modification: incubation time in freshly satu- cheeks, here alternating with black spots. The ven- at rated Ba(OH) solution was 5 min. 60°C. They were then 2 tral dark and roundish in spots are usually large rinsed in 1% acetic acid solution for 5 min. The slides obtained males and small and more elliptically shaped in fe- were incubated in 2 x SSC at 60°C for 30 min. Finally, prepara-

distilled water and stained with 3% Giemsa males. The ventral colour continues tions were rinsed in background in phosphatebuffer, pH 6.8. Giemsa stained preparations were Juveniles show similar but over the guiar region. a photographed with a Zeiss Universal microscope using Kodak brighter colour than adults. In all juvenile speci- Imagecapture AHU microfilm. mens studied, a conspicuous orange-red dorsalline, All specimens examined and their localities are listed in the ventral coloura- dorsal hues, and a yellowish Appendix. green tion are present.

During the aquatic phase, the male dorsal col-

Results ouration changes drastically. The black spots get

lighter, becoming brown or bronzed with broad

External morphology blackish dots in the middle. Occasionally, portions

of the terrestrial-phase spots become mixed with

- which sometimes Redescription of T. m. pygmaeus. Large head, the rest of the dorsal colouration,

show bronze back- longer than wide, gradually sharpened, ending as cupreous or tones on a green

Small of this be in abruptly in a narrow, blunt nose. eyes ground (an example pattern can seen

Prominent fold. 1980: Some individuals placed in a forward position. guiar Andrada, plate 33). may

Elongated body with long, thin legs. Digits long, become completely dark and melanic in appear-

females show especially in males. Toe webbing absent. Relative- ance; may evenly-spread green-grey ly short tail, its cross-section being almost cylin- hues. The inferior caudal crest is black in the male,

lower drical at the base and progressively compressed with some white dots. In the female, the part towards the tip. Finely granulated skin, ridged in of the crest usually shows orange-yellowish, or the guiar region, with large granules unevenly greyish tones.

basic statisti- spread over the dorsal surface. Well developed To complete the description, some

related total labial folds during the aquatic phase. During the cal parameters to length, snout-vent

distance between forelimbs and breeding period the males have a relatively low length, hindlimbs, linear dorsal crest, which continues along the tail forelimb length, third finger length, hindlimb without notable inflexion in the third toe length, head width, and head interruption or any length,

indicated for each in Table I. pelvic area. length are sex

dorsal colour During the terrestrial phase the pat-

of black of T. and T. tern consists of an irregular pattern spots Comparison m. marmoratus m. pyg-

These - The of T. m. on a green background. spots can create a maeus. comparative analysis pyg-

from Cádiz and T. from reticulum or a more or less wide bandrunning along maeus m. marmoratus the side of the body. In the females, the dorsal mid- Cantabria shows the following diagnostic features line continuous for T. appears as a red-orange stripe. m. pygmaeus:

- Size: T. is smaller in size than T. Males, however, have a line with alternating m. pygmaeus orange-yellow and black narrow bands instead of m. marmoratus (Table I), whether using natural the stripe. The dorsal part of the tail shows the same log-transformed (Lemen, 1983) or non-trans-

formed data. all the variables with the colour as the dorsum. Ventrally, the colourationis In studied,

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exception of head length, there are highly signifi- pearance, rather differentfrom the marbled pattern

cant differences (p < 0.01) between the two groups typical of Cádiz populations. Dotted specimens

of populations. Although all these measurements generally show rounder snouts and shorter digits.

couldbe used as diagnostic feature of either group, In thePortuguese population of Portocovo the ven-

the tral white we propose snout-vent length as simplest one, dots are limited to the periphery of the

because there is hardly any overlap between the two belly. In some populations from the provinces

groups studied, and it is easily measured both in Madrid, Sevilla, and Cáceres, the white dots have

museum specimens and in the field. low densities. The distributionof ventral white dots

Results from the discriminant does not to follow a posteriori analy- seem any obvious geographical

sis of the log-transformed data set from the 315 trend.

show for specimens measured, effective separation Slight variations were observed in the dorsal

the two subspecies. Histograms of discriminant colouration of T. m. marmoratus with big round

demonstrate clear of the in the scores a separation subspe- spots prevailing populations from La Rioja.

cies with only slight overlap (Fig. 5). Results for the We have also observed conspicuous individual

original data set are practically identical to those differences in ventral background colouration;

for the transformed data. specimens showing light marbled areas have occa-

- Ventral colouration: in typically creamy yellow sionally appeared in some populations from

T. m. pygmaeus, marked to a greater or lesser ex- Salamanca and Madrid.

tent with large black spots and small white ones. In

T. m. marmoratus the ventral colourationis black- Cytogenetic analysis ish with fine white dots and, as a consequence, the

black dots can rarely be seen. The white dotsof T. Triturus marmoratus has 24 biarmed chromosomes

and of of m. pygmaeus are relatively larger more irregu- decreasing length, all them sub- or metacen-

lar than those of T. m. marmoratus; usually they tric. As Mancino & Nardi (1971), Callan & Lloyd

are concentrated at the flanks, Sims especially covering (1975), et al. (1984) and many other authors

large surfaces, whilein T. m. marmoratus the white noticed, this species shows a fixed heteromorphism

dots all the ventral in chromosome are spread over region. 1, with the long arm of one of the

- Dorsal crest: relatively low, linear and without chromosomes being considerably longer than that

inflexion in T. of any pelvic m. pygmaeus; relatively the other chromosome (Fig. 2).

high (especially in the centralbody area), crenulat- All individuals except those from Doñana

ed and with evident inflexionin an the pelvic region presented the following C-bands pattern:

in T. m. marmoratus. The chromosomes of T. marmoratus have large

males and females of and distinct centromeric Representative T. m. pyg- very C-bands, with the ex-

maeus populations (Algeciras Bay, Cádiz) and T. ception of chromosome 9 which has small centro-

meric C-bands m. marmoratus populations (Cantabria) are shown (Fig. 2). All chromosomes have

in 1. C-bands in both Fig. pericentric arms, except the long

arm of pair 5 and the short arm of pair 12. Intersti-

variation. - Some tial thin C-bands inthe of chromo- Intrasubspecific intrasubspecific appear long arm

variation was observed in features such as size, dor- some 7 and in the short arms of chromosome 4.

sal colourationand the extent of the ventral white Terminal heterochromatin may be detected on

dotting. T. m. pygmaeus populations from the the long arms of chromosomes 3, 4, 5, 8, and 9.

south of the province of Huelva (Doñana, El Ro- Pair 4 constitutes the sexual chromosomes. Males

show smaller size cío) clearly a than those from the show a C-band heteromorphism at the ends of the

province of Cádiz. Populations from the provinces long arms. Chromosome Y has more terminal het-

Ciudad Cáceres, Real, Toledo, and Madrid show a erochromatin than chromosome X (Fig. 2). The

peculiar dorsal pattern, consisting of round black long arms of chromosome 1 show two C-band het-

spots on a greenbackground; these spots, generally eromorphisms in all individuals, irrespective of sex. abundant and never fused, them a dotted give ap- A large heterochromatic region, extending from

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Fig. 1. Representative males (a) and females (b) from T. m. marmoratus (Cantabria, Spain) and T. m. pygmaeus (Algeciras Bay, Cádiz

Province, Spain) (c, d).

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Fig. 2. C-banded karyotype and idiogram representing the distribution of C-bands in the mitotic chromosomes of T. marmoratus.

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Fig. 3. C-banded karyotype and idiogram representing the distribution of C-bands in the mitotic chromosomes of a T. marmoratus in- dividual from Doñana. Chromosome 4X shows a double pericentric band in the short arm.

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near the centromere to near the end, is present on Peninsula (Rivas Martínez, 1973; 1982), the distri-

these arms. This region is more extensive on the butionof T. m. marmoratus in the Iberian Peninsu-

the long chromosome 1A than on short chromo- la seems to be coincident with the extension of the

IB. C-band of chromosome Eurosiberian the some The pericentric region and Supramediterranean

close whereas in chromo- bioclimatic Mediterranean 1A is to the centromere, area of the region. The

some IB it is close to the large heterochromatic Supramediterranean area is characterized by an an-

region (Fig. 2). These results are in agreement with nual mean temperature of 8 to 12°C and mean

described those by Sims et al. (1984). minimum and maximum temperatures in the col-

The population from Doñana is unique in that all dest month of -3 to 0°C and 3 to 8°C, respectively

individuals show two small but fixed differences on (Rivas Martínez, 1982). There are some exceptions

their C-banded chromosomes. These differences in the mountains, where they reach the Oromediter-

absence of the are: the pericentric band in the long ranean area, and in the Ebro and Duero valleys as

of chromosome 12 and the of well in arms presence a as some zones at the southern slopes of the

double pericentric band in the short arms of chro- Sistema Central, where they reach the Mesomedi-

mosome X (Fig. 3). terranean bioclimatic area.

The distributionof T. m. pygmaeus is mostly in-

cluded within the Mesomediterraneanbioclimatic Distribution area, characterized by an annual mean temperature

of 12 to 16°C and mean minimum and maximum One of the main problems in studying Iberian temperatures in the coldest month of 0 to 5°C and populations of T. marmoratus has been the dif- 8 to 13°C, respectively (Rivas Martínez, 1982). ficulty in assigning individuals to one of the two

There are also populations of this taxon in the Ther- subspecies, due to the abbreviated original descrip- momediterraneanarea of the Guadalquivir valley tion of T. m. pygmaeus by Wolterstorff (1905). and the southern coast of Portugal and in the According to the diagnostic features described

Supramediterranean area of some central and above, we assigned all specimens examined to one southern mountains. of the two subspecies (Fig. 4) and the following dis- The ecological characteristics of the breeding tribution pattern was inferred:

sites used by both T. m. marmoratus and T. m. pyg- The distributionof T. m. pygmaeus ranges along

maeus are difficult to generalize as a consequence the southern half of the Iberian Peninsula (except

of the showed both taxa the in the great variety by along eastern areas, where T. marmoratus seems to

be absent), reaching at some places the southern

slopes of the Sistema Central, which runs across the

central region of the Iberian Peninsula. At one

point (Puerto de Malagón, El Escorial, in the

of province Madrid, Spain), the northern range of

T. m. pygmaeus seems to reach beyond the Sistema

Central.

The southern distributionallimits of T. m. mar-

moratus reach beyond the Sistema Central and

cover the southern slopes of mountains in the fol-

lowing three zones: the eastern part of the Sierra de

Guadarrama, in the province of Madrid (Spain);

Sierra de Gata, in the province of Cáceres (Spain); and Serra daEstrela, in the province of BeiraBaixa

(Portugal). of localities Fig. 4. Map of material examined. T. m. marmora- Comparing those distributions with the regions localities and lo- tus are representedby squares T. m. pygmaeus and bioclimatic areas proposed for the Iberian calities by dots.

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the only existing records referring to Cádiz and

Huelva (Salvador, 1974; Collado et al., 1976) to

their contact zone with the southernmost limits

known for T. m. marmoratus (Dorda & Esteban,

1986), along the Central Range Mountains (Sistema

in Central), as Macgregor et al. (1990) represented

their Fig. 1.

In the zones where the southern distributional

rangeof T. m. marmoratus reaches beyond the Sis-

the tema Central, a contact area between two

subspecies is to be expected. However, we have so

far failed in our search for individuals with inter-

mediate characters, and no areas with strictly sym-

patric populations have yet been found. The closest

populations with no apparent geographic barriers

betweenthem are separated by 6 km in the Sierra de

in Sierra de Guadar- Gata zone, and by 20 km the

rama zone (Fig. 4).

This new distribution is in line with the geo-

graphical dataarising from the biochemical analy-

sis given by J.W. Arntzen, in Dorda & Esteban

(1986) (Nei's Distance D = 0.25 between El Berrue-

Fig. 5. Histogram of canonical variates for male (a) and female co and Villalba populations), and Macgregor et al.

(b) T. marmoratus subspecies generatedfrom the two-group dis- (1990) (Nei's Distance D = 0.19 between Balenyá criminant analysis, conducted using natural-log transformed and Lisboa populations), because, according to our data. data, the populations from Balenyá (Barcelona,

Spain) and El Berrueco (Madrid, Spain) belong whole area occupied. In general, both use small while from Villalba to T. m. marmoratus, those in forested man-made ponds rivers, ponds areas, (Madrid, Spain) and Lisboa (Portugal) belong to for cattle in grassland areas, troughs, quarries, the subspecies T. m. pygmaeus. ditches in mountainponds, etc. agricultural lands, Nei's distances shown by these authors indicate

In the border area of both taxa at the Sistema Cen- clear differentiation between T. m. marmoratus tral they use similar ponds, locatedon a granite san- and T. m. pygmaeus. In the present study this is dy soil with different degrees of vegetational cover. dif- reflected by some remarkable morphological In those T. marmoratus to be areas, m. seems ferentiation in features associated with reproduc- found most frequently in the Supramediterranean tion, and in size. This result is confirmed by the dis- area, corresponding with the potential oak forest criminant analysis carried out a posteriori (Fig. in of Quercus pyrenaica, and T. m. pygmaeus 5). However, the interpopulational variability ob- the Mesomediterranean or Supramediterranean, served in many other features, considered not to be corresponding to the potential forest of Quercus diagnostic, deserves further analysis. Possibly geo- ilex. graphical trends in the observed patterns will be

revealed.

The cytogenetic analysis does not show any in-

Discussion trasubspecific differentiation, with the exception of

the Doñana population. From a chromosomal

distributional of view the characteristics shown this As a consequence of these results, the point by spe- map of T. m. pygmaeus should be extended from cies are among the most conservative within the

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Triturus Recent data the Calderón & M. 1976. Datos sóbrela fauna genus (Herrero, 1985). on Collado,E.,J. Pérez,

de anfibios del Bajo Guadalquivir. Doñana, Acta Vertebrata, cytogenetics of T. marmoratus seems to have led to 3(1): 5-17. some confusion, perhaps because different idio- Dixon, W.J. (ed.), 1983. BMDP Statistical Software: 1-733 C-banded grams were drawn from the same prepa- (Univ. California Press, Berkeley). rations Sims et al., 1984 and et (cf. Macgregor al., Dorda, J. & M. Esteban, 1986. Evolution and subspeciation of

1990, with thepresent study). We conclude thatthe Iberian Triturus marmoratus. In: Z. Rocek (ed.), Stud. Her-

morphological and biochemical differentiation petol., Prague: 159-163.

Herrero, P., 1985. Estudios citogenéticos en las especies del achieved by T. m. pygmaeus populations is not género Triturus de la Península Ibérica: 1-147 (Ph. D. The- paralleled at the chromosomal level. sis, Universidad Autónoma de Madrid). As strict has been and no sympatry yet observed, Lemen, C., 1983. The effectiveness of methods of shape analy-

individuals with intermediate be- sis. an appearance Fieldiana, Zool., 1343: 1-17.

H.C., S.K. Sessions & J.W. 1990. An in- tween both subspecies have not been found, it is Macgregor, Arntzen,

tegrative analysis of phylogeneticrelationships among newts necessary to study the potential areas of contact in ofthe Triturus genus (family Salamandridae),using compara- more detail. Studies in the areas of Avila, Cáceres, tive biochemistry, cytogenetics and reproductive interactions. and Madrid (Spain), and in Beira Baixa provinces J. evol. Biol., 3: 329-373.

and Estremadura (Portugal) would be particularly Mancino, G. & I. Nardi, 1971. Chromosomal heteromorphism

and female heterogamety in the marbled newt Triturus mar- interesting, so as to ascertain the degree of differen-

moratus (Latreille, 1800). Experientia, 27: 821-822. tiation and isolation achieved by the two forms. Mertens, R. & L. Millier, 1928. Die Amphibien und Reptilien The potential existence of a narrow contact zone, Europas. Zweite liste. Abh. senckenb. naturf. Ges., 41:1 -62.

with flow or the gene absent very low, poses ques- Avance Rivas-Martínez, S., 1973. sobre una síntesis corológica

tion whether T. should be m. pygmaeus assigned de la Península Ibérica, Baleares y Canarias. Anal. Inst. Bot.

33: 167-188. specific status. Cavanilles, Rivas-Martínez, S., 1982. Mapa de las series de vegetación de

Madrid (Diputación de Madrid, Madrid).

Acknowledgements Salvador, A., 1974. Guía de los anfibios y reptiles de España:

1-237 (1CONA, Madrid).

We are indebted to I. Esteban, A. Múgica, and F. Barrios for Sims, S.H., H.C. Macgregor, P.S. Pellat & H.A. Horner, 1984.

their valuable cooperationin the fieldwork; to Dr. J.W. Arntzen Chromosome 1 in crested and marbled newts (Trituras). An

for sendingus some of his material and for helpful suggestions; extraordinary case of heteromorphism and independentchro-

to Prof. E.G. Crespo who gave us access to the collections of mosome evolution. Chromosoma, 89: 169-185.

Museu Bocage; to L. van Tuyl for providing us with material Sumner, A.T., 1972. A simple technique for demonstrating cen-

from the Zoölogisch Museum, Amsterdam; to the "Agencias de tromeric heterochromatin. Exp. Cell Res., 75: 304-306.

Medio Ambiente" from the "Comunidades Autónomas" of Wolterstorff, W., 1905. Zwergformen der Palàarktischen Uro-

Andalucía, Asturias, Castilla - La Mancha and Madrid for delen. C. r. 6e Congr. int. Zool., Berne: 258-263.

authorizingus to capture specimens from areas not represented Zar, J. H., 1984. Biostatistical analysis (2nd ed.): 122-149

in museum collections; to C. Hosie, A. Smidthson, and L. (Prentice Hall, Inc., New Jersey).

García-Hidalgofor helpfullinguistic suggestions.This work was

supportedby CICYT Project PB 88-0010 and the CSIC - British

Council Cooperative Project (Spain-Great Britain) HB-180. Received: 27 February 1992

Revised: 27 October 1992

References Appendix

Guía de de Andrada, J., 1980. campo los anfibios y reptiles de Material examined

la Península Ibérica: 1-51 (Omega, Barcelona).

Busack, S.D., B.G. Jericho, L.R. Maxson & T. Uzzell, 1988. (a) Museum collections:

ofsalamanders The MNCN = Museo Evolutionaryrelationships in the genus Tritu- acronyms used are the following: Na-

ras: The view from immunology. Herpetologica, 44: 307- cional de Ciencias Naturales (Madrid, Spain); MBL = Museu

316. Bocage (Lisboa, Portugal); ZMA = Zoölogisch Museum Am-

H.G. & L. 1975. ofthe sterdam The = British Callan, Lloyd, Working maps lampbrush (Amsterdam, Netherlands); BMNH

of Amphibia. In: R.C. King (ed.), Handbook of genetics, 4: Museum (Natural History) (London,U.K.); ct indicates that the

57-77 (Plenum, New York). specimen was cytogenetically analyzed.

Downloaded from Brill.com09/23/2021 11:57:05AM via free access Bijdragen tot de Dierkunde, 63 (1) - 1993 13

T. m. marmoratus: : La Granja: 3 specimens, MNCN 783-85.

Valladolid: Valladolid: 2 specimens, MNCN 838-39.

PORTUGAL: Cabeço do Infante: 3 specimens, MNCN Zamora: Fuentes de Ropel: 15 uncatalogued specimens (et).

9777-79; Freixadas-Alverca: 1 specimen, MBL. Zaragoza: Tarazona: 3 uncatalogued specimens (et).

Beira Alta: Serra da Estrela: 1 specimen, MNCN 2425; Vilar

Formoso: 1 ZMF 7281. T. specimen, m. pygmaeus:

Beira Baixa: Castelo Branco: 3 specimens, MNCN 9783-85.

Douro Litoral: Valongo: 1 specimen, ZMF 7682. PORTUGAL: Beira Baixa: Castelo Branco - Escalos de Baixo:

Minho: Braga: 2 specimens, MNCN 777-78; Geres - Braga: 5 1 specimen, MBL.

specimens, MBL; Bom Jesus (Braga): 3 specimens, MBL. Estremadura: Lagoas do Arrimai - Porto de Mos: 4 specimens,

Tras-os-Montes: Serra da Nogueira (sur de Bragança): 1 MBL; Sintra: 2 specimens, MBL; Sintra: 2 specimens, BMNH

specimen, MBL; Rebordaos, Serra da Nogueira, 1 specimen, 189632668; Palace Grounds of Quiluz, 1 specimen, BMNH

MBL. 18767767.

Algarve: Tavira, N 270 km 58.8: 3 specimens, MBL.

SPAIN: Asturias: Oviedo: 10 uncatalogued specimens (et), 1 Alto Alemtejo: Portalegre - Castelo de Vide: 5 specimens, specimen, MNCN 767, 1 specimen, MNCN 9593; Posada: 1 MBL; Estremoz: 2 specimens, MNCN 9780-81; Vimiciro: 3 specimen, MNCN 847; La Muela - Pola de Lena: 3 specimens, specimens, MBL; Cabeçao: 2 specimens, MBL; Serra de Sâo

MNCN 9595-97; Pola de Siero: 5 uncatalogued specimens (ct), Mamede (Portalegre): 1 specimen, MBL.

2 specimens, MNCN 11197-98; Gijón: 16 uncatalogued speci-

mens (ct), 10 specimens, MNCN 790-99, 5 specimens, MNCN SPAIN: Avila: Piedralaves - Mijares: 3 uncatalogued speci-

819-23; Asturias: 1 specimen, MNCN 766. mens (ct), 4 specimens, MNCN 9581-84.

Avila: Avila: 4 specimens, MNCN 2402-05. Cáceres: : 9 uncatalogued specimens (ct),

Barcelona: Barcelona: 1 specimen, ZMF 7637. 2 specimens, MNCN 9603-04; Trujillo: 2 specimens, MNCN

Burgos: Bujedo: 1 specimen, MNCN 757; Pancorbo: 8 speci- 9782, 15339; : 1 specimen, MNCN 2426; Villa Real mens, MNCN 11112-19. de San Carlos: 5 uncatalogued specimens (ct), 2 specimens,

Cantabria: Santillana del Mar: 10 uncatalogued specimens (ct), MNCN 13071, 15334.

6 specimens, MNCN 11189-94, 5 specimens, MNCN 13072-76, Cádiz: Los Barrios: 6 specimens, MNCN 15313-18 (ct), 7 speci-

1 specimen, MNCN 13265, 1 specimen, MNCN 15337, 3 speci- mens, MNCN 15319-21, 15331-32, 15949-50, 1 uncatalogued

MNCN mens, 16095-97; Oreña: 9 specimens, MNCN 13077-85, specimen; San Roque: 1 specimen, MNCN 15948.

6 specimens, MNCN 16089-94; Lago Revilla: 3 specimens, Ciudad Real: Daimiel: 4 specimens, MNCN 1014-16, 1018;

MNCN 2433-35; Embalse del Ebro - Reinosa: 2 uncatalogued Ruidera: 2 specimens, MNCN 845-46; Río Tablillas: 1 specimen, specimens (ct), 1 specimen, MNCN 15336; Santander: 3 uncata- MNCN 15932. logued specimens (ct), 1 specimen, MNCN 16098. Córdoba: Venta del Charco: 10 specimens, ZMA 7677, 4 speci-

Gerona: Tosa: 5 uncatalogued specimens (et). mens, ZMA 8046.

La Coruña: La Coruña: 1 specimen, MNCN 758, 1 specimen, Granada: Zafarraya: 25 specimens, MNCN 15299-306, 15335,

MNCN 800, 13 specimens, MNCN 806-18; Los Castros, 12 15951-55, 11 uncatalogued. specimens, MNCN 824-35; Santa María de Ois: 1 specimen, Huelva: Doñana: 8 uncatalogued specimens (et); 5 specimens,

MNCN 15338. MNCN 11217, 15333, 16086-88.

La Rioja: Logroño: 2 specimens, MNCN 804-05, 1 specimen, Jaén: Jaén: 1 specimen, MNCN 11195.

MNCN 9406; Valgañón: 1 specimen, MNCN 16066. Madrid: Hoyo de : 2 specimens, MNCN 9601-02, 1

León: Foncebadón: 2 uncatalogued specimens (et); León: 2 specimen, MNCN 11199, 6 specimens, MNCN 15307-12, 7 specimens, MNCN 2427-28. specimens, MNCN 15324-30, 10 specimens, MNCN 15956-65, 3

Madrid: El Berrueco: 8 uncatalogued specimens (et), 2 speci- specimens, MNCN 16077-79; Villalba: 4 uncatalogued speci-

2 mens, MNCN 9589-90, 2 specimens, MNCN 9591-92, 1 speci- mens (et), specimens, MNCN 9570-71, 2 specimens, MNCN men, MNCN 9594, 9 specimens, MNCN 13062-70, 2 specimens, 13053-54; Alpedrete: 2 uncataloguedspecimens (et); Pantano de

MNCN 15340-41, 6 specimens, MNCN 16068-73; El Paular: 3 San Juan: 1 specimen, MNCN 9585; Torrelodones: 10 uncata- specimens, MNCN 2468-70, 3 specimens, MNCN 2472-74, 3 logued specimens (et), 2 specimens, MNCN 9586-87, 1 speci-

2 MNCN specimens, MNCN 2476-78; La Cabrera: specimens, men, MNCN 15343,3 specimens, MNCN 16074-76;Chapinería:

15322-23, 1 specimen, MNCN 15344, 3 specimens, MNCN 3 specimens, MNCN 9575-77; Navalagamella: 3 specimens,

15944-46. MNCN 9572-74, 7 specimens, MNCN 13055-61; El Pardillo: 4

Navarra: Pamplona: 2 specimens, MNCN 836-37. specimens, MNCN 786-89.

Orense: Orense: 4 specimens, MNCN 779-82. Sevilla: Gerena: 5 uncatalogued specimens (et), 2 specimens,

Pontevedra: Vigo: 1 specimen, MNCN 2432; Tuy: 2 specimens, MNCN 9599-9600, 11 specimens, MNCN 13085-95, 2 speci-

MNCN 775-76; Pontevedra: 1 specimen, MNCN 768; Moaña: 6 mens, MNCN 16080-81; Sevilla: 2 specimens, MNCN 2422-23. uncatalogued specimens (et), 1 specimen, MNCN 16067. Toledo: Nombela: 3 specimens, MNCN 9578-80; Navalcán: 1

Salamanca: Montemayor del Río: 1 specimen, MNCN 9588; specimen, MNCN 2421; Pelahustán: 8 uncatalogued specimens

Robleda: 6 specimens, MNCN 769-74. (et), 7 specimens, MNCN 11218-24.

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(b) Sites not mentioned in the former list, where individuals were T. m. pygmaeus:

observed and later freed in situ:

SPAIN: Avila: Mijares, , La Adrada.

Cáceres: Navalmoral de la Lanchares de Alcán- T. m. marmoratus: Mata, (Valencia

tara), Logrosán, , , ,

SPAIN: Avila: Avila. Pozuelo de Zarzón.

Cáceres: Hervás, , , Cádiz: Alcalá de los Gazules.

Cilleros, Santibañez el Alto. Ciudad Real: El Viso del Marqués.

Cantabria: Noja. Huelva: El Rocío.

Gerona: Roses. Madrid: Puerto de Malagón, El Escorial, Zarzalejo, Val-

La Coruña: Finisterre. demorillo, , Pelayos de la Presa, de los

Madrid: Peñalara, Rascafría. Vidrios, Colmenar Viejo, Becerril, Cerceda, Valdemaqueda,

Pontevedra: Cutián. Colmenar del Arroyo.

Salamanca: El Cabaco, El Maíllo, Candelario. Toledo: San Pablo de los Montes.

Segovia: El Espinar, Villacastín.

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