Caryologia

International Journal of Cytology, Cytosystematics and Cytogenetics

ISSN: 0008-7114 (Print) 2165-5391 (Online) Journal homepage: https://www.tandfonline.com/loi/tcar20

Chromosomal Studies in Brazilian Anurans

Maria Nazareth Rabello

To cite this article: Maria Nazareth Rabello (1970) Chromosomal Studies in Brazilian Anurans,
Caryologia, 23:1, 45-59, DOI: 10.1080/00087114.1970.10796362

To link to this article: https://doi.org/10.1080/00087114.1970.10796362

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 CHROMOSOMAL STUDIES IN BRAZILIAN ANURANS *

MARIA NAZARETH RABELLO

Servi\o de Genetica, Instituto Butantan, S. Paulo, Brazil

Received: 12th August 196,

INTRODUCTION

The comparative study of the karyotypes of close species may help our
understanding of the mechanisms of chromosomal evolution relevant to spe
ciation. It may also clarify phylogenetic relationships.
Anurans are abundant, varied and widespread. This and the diversity
of their karyotypes make them particularly suited to cytogenetic research.
In the present paper, the karyotypes and male meiosis of nine species
of Hylidae and of one Microhylidae are described. Including those, 186 spe-
cies distributed among fifteen families of Salientia have their diploid number
known at present (Table I).

TABLE I
Summary of karyotipic data in Salientia.

Species 2n n (a) Author

Family Ascaphidae
Ascaphus truei 44 MORESCAI.CHI 1967
Family Leiopelmatidae
Leiopelma hochstetteri 'i' 34 MoRESCALCHI 1968
'i' 23
Family Pipidae
Pipa parva 30 MoRESCALCHI 1968
Family Xenopidae
Hymenochirus boettgeri 24 MoRESCALCHI 1968
Xenopus laevis 36 18 WICKBOM 1945
Family Pelobatidae
Pelobates cultripes 26 MORESCALCHI 1968
Pelobates fuscus 26 13 WICKBOM 1945, 1949
Pelodytes puncta/us 6(MI) BATAILLON 1910
Pelodytes puncta/us 24 110RESCALCHI 1968

* This work was supported by US. PHS-NIH Research Grant GM-14577-03 from the Na-
tional Institute of General Medical Sciences, by the Fundo de Pesquisas do Instituto Butantan
and by the Conselho Nacional de Pesquisas.

[Caryologia, Vol. 23, n. 1, 1970

46 RABELLO

(Table I cont.)
Species 2n n( &) Author

Scaphiopus bombifrons 13 CoPE and HuGHES 1962

Scaphiopus couchi 26 ~ORESCALCHI 1968
Scaphiopus hammondi 13 CoPE and HuGHES 1962
Scaphiopus holbrooki holbrooki 13 WITSCHI 1933
26 TING and PRICE 1950
Scaphiopus holbrooki hurteri 26 WASSERMAN and BOGART 1968
Family Discoglossidae
Alytes obstetricans 32 16 }ANSSENS and WILLENS 1909
Alytes obstetricans 36 18 WICKBOM 1949
Alytes obstetricans 38 ~ORESCALCHI 1968
Bombina bombina 24 12 WICKBOM 1949
~ORESCALCHI 1965
Bombina variegata 24 12 WICKBOM 1949
~ORESCALCHI 1965
Discoglossus pictus 28 WICKBOM 1945, 1949
Family Bufonidae
Bufo americanus 11 WITSCHI 1943
Bufo arenarum 22 11 SAEZ et al. 1934
BIANCHI and LAGUENS 1964
Bufo boreas 22 SANDERS and CRoss 1964
Bufo brauni 20 BoGART 1968
Bufo bufo 22 WICKBOM 1945, 1949
Bufo bufo japonicus 22 11 IRIKI 1929
~INOUCHI and IRIKI 1931
Bufo calamita 12 BATAILLON 1910
Bufo calamita 22 11 STOHLER 1927
WICKBOM 1945, 1949

Bufo canorus 11 WITSCHI 1933

Bufo carens 22 BoGART 1968
Bufo cognatus 22 11 ~ORESCALCHI and (;ARGIULO 1968
Bufo compactillis speciosus 22 HAYES 1933
Bufo crucifer 22 11 BE«;:AK ~.L. 1967, 1968
Bufo fowleri 11 WITSCHI 1933
Bufo gariepensis 22 BoGART 1968
Bufo garmani 20 BoGART 1968
Bufo garmani 20 10 ~ORESCALCHI and (;ARGIULO 1968
Bufo granulosus d'orbignyi 22 SAEZ and BRUM 1960
Bufo granulosus d'orbignyi 22 11 BE«;:AK ~.L. et al. 1969
Bufo gutturalis 20 BoGART 1968
Bufo hemiophrys 22 11 ~ORESCALCHI and (;ARGIULO 1968
Bufo ictericus 22 11 BE«;:AK ~.L. 1967, 1968
Bufo latifrons 20 BoGART 1968
Bufo lentiginosus 24 12 KING 1901
Bufo lentiginosus 11 WITSCHI 1933
Bufo mauritanicus 22 BOGART 1968
Bufo melanostictus 22 NATARAJAN 1958
Bufo melanostictus 22 11 ~ORESCALCHI and (;ARGIULO 1968
Bufo paracnemis 22 11 BE«;:AK ~.L. 1967, 1968
Bufo quercicus 11 WITSCHI 1933
Bufo raddei 22 11 SATO 1936
Bufo rangeri 20 BoGART 1968
Bufo rangeri 20 10 ~ORESCALCHI and (;ARGIULO 1968
Bufo regularis 11 WICKBOM 1949
Bufo regularis 20 BoGART 1968
BECKERT and DoYLE 1968
 CHROMOSOMAL STUDIES IN ANURANS 47

(Table I cont.)
Species 2n n(~) Author

Bufo regularis 20 10 MoRESCALCHI and GARGIULO 1968

Bufo rosei 22 BoGART 1968
Bufo « Sachalinensis » 22 11 MAKINO 1930, 1932
Bufo spinulosus 22 11 MORESCALCHI and GARGIULO 1968
Bufo valliceps 22 HAYES 1933
Bufo viridis 22 11 STOHLER 1926; BECCARI 1926;
GALGANO 1933; WICKBOM 1945
Bufo vulgaris 8-10~ (MI) CARNOY and LEBRUN 1900
Bufo vulgaris 8 ~(Mil) LEBRUN 1901
Bufo vulgaris 1824 DELLA VALLE 1907
Bufo vulgaris 8-9(MI) BATAILLON 1910
Bufo vulgaris 22 11 STOHLER 1927; GALGANO 1933;
WICKBOM 1945
Bufo vulgaris 22-24 POSKA-TEISS 1933
Bufo woodhousei 22 HAYES 1933
Family Leptodactylidae
Calyptocephalella gayi 26 BRUM-ZORRILLA and SAEZ 1968
Crossodactylus gaudichaudii 22 BRUM-ZORRILLA and SAEZ 1968
Crossodactylus grandis 26 13 BEc;:AK M_L_ 1967, 1968
Crinia signifera 24 MoRESCALCHI et al. 1968
Crinia sloanei 24 MoRESCALCHI et al. 1968
Cycloramphus asper 26 13 BEc;:AK M.L. et al. (In press)
Cycloramphus dubius 26 13 BEc;:AK M.L. et al. (In press)
Cycloramphus fuliginosus 22 BRUM-ZORRILLA and SAEZ 1968
Eleutherodactylus discoidalis 22 BRUM-ZORRILLA and SAEZ 1968
Eleutherodactylus guentheri 22 11 BEc;:AK M.L. 1967, 1968
Eleutherodactylus guentheri BRUM-ZORRILLA and SAEZ 1968
Eleutherodactylus ranoides 22 BRUM-ZORRILLA and SAEZ 1968
Elosia aspera 26 BRUM-ZORRILLA and SAEZ 1968
Elosia lateristrigata 26 BRUM-ZORRILLA and SAEZ 1968
Eupemphix nattereri 22 11 BEc;:AK M.L. 1967, 1968
Engystomops pustulosus 22 MORESCALCHI and GARGIULO 1968
Eupsophus nodosus 22 BRUM-ZORRILLA and SAEZ 1968
Leptodactylus bufonius 22 BRUM-ZORRILLA and SAEZ 1968
Leptodactylus chaquensis 22 BRUM-ZORRILLA and SAEZ 1968
Leptodactylus laticeps 22 BRUM-ZORRILLA and SAEZ 1968
Le ptodactylus ocellatus 22 SAEZ and BRUM 1960
Leptodactylus ocellatus 22 11 BEc;:AK M.L. 1967, 1968
Leptodactylus pentadactylus 22 BRUM-ZORRILLA and SAEZ 1968
MoRESCALCHI and GARGIULO 1968
Leptodactylus prognathus 22 BRUM-ZORRILLA and SAEZ 1968
Leptodactylus sibilatrix 22 11 BEc;:AK M.L. et al. 1969
Limnodynastes peroni 24 RoBINSON and STEPHENSON 1967
MORESCALCHI et al. 1968
Limnodynastes fletcheri 24 MoRESCALCHI et al. 1968
Physalaemus biligonigerus 22 BRUM-ZORRILLA and SAEZ 1968
Physalaemus cuvieri 22 11 BEc;:AK M.L. et al. 1969
Physalaemus fuscomaculatus 22 11 BEc;:AK M.L. et al. 1969
Physalaemus gracillis 22 BRUM-ZORRILLA and SAEZ 1968
Pseudopadudicola ameghini 20 10 BEc;:AK M.L. 1967, 1968
Pseudopaludicola falcipes 22 SAEZ and BRuM 1960
Pseudopaludicola falcipes 18 9 BEc;:AK M.L. 1968
Pseudopaludicola falcipes 18,20 9,10 BATISTIC et al. 1969
Pseudophryne bibronii 24 MoRESCALCHI et al. 1968
Pleurodema cinera 22 BRUM-ZORRILLA and SAEZ 1968
Pleurodema nebulosa 22 BRUM-ZORRILLA and SAEZ 1968
Pleurodema tucumana 22 BRUM-ZORRILLA and SAEZ 1968
48 RABELLO

(Table I cont.)
Species 2n n( ~) Author

T elmatobius (group marmoratus) 22 BRUM-ZORRILLA and SAEZ 1968

Thoropa miliaris 26 BRUM-ZORRILLA and SAEZ 1968
U peroleia marmorata 24 MoRESCALCHI et al. 1968
Family Ceratophrydidae
Ceratophrys calcarata 26 MoRESCALCHr 1967
Ceratophrys dorsata 104 52 BEC,:AK M.L. et al. 1967
Ceratophrys ornata >80 SAEZ and BRUM 1959
Macrogenioglotus allipioi 22 11 BEC,:AK M.L. et al. (In press)
Odontophrynus americanus 42,44,50 SAEZ and BRUM 1966
Odontophrynus americanus 44 22 BEC,:AK M.L. et al. 1967
Odontophrynus carvalhovi 22 11 BEc;:AK M.L. et al. (In press)
Odontophrynus cultripes 22 SAEZ and BRUM 1966
Odontophrynus cultripes 22 11 BEC,:AK M.L. et at. 1967
Odontophrynus occidentalis 22 SAEZ and BRUM 1966
Odontophrynus occidentalis 22 11 BEC,:AK M.L. et al. 1968
Stombus appendiculatus 22 11 BEc;:AK M.L. et al. 1968
Stombus boiei 22 11 BEC,:AK M.L. et al. 1968
Zacchaenus parvulus ( =Oocormus
microps) 24 12 BEC,:AK M.L. et al. (In press)
Family Pseudidae
Pseudis paradoxus platensis 24 12 BEC,:AK M.L. 1967, 1968
Family Hylidae
Acris crepitans 22 11 BuSHNELL et al. 1939
Acris gryllus 22 11 BUSHNELL et al. 1939
Anotheca coronata 12 DuELLMAN and CoLE 1965
Diaglena reticulata 12 DuELLMAN and CoLE 1965
Diaglena spatulata 12 DuELLMAN and CoLE 1965
Fritziana goeldii 26 13 BEC,:AK M.L. 1967, 1968
Hyla albomarginata 24 12 BEC,:AK M.L. 1967, 1968
Hyla albopunctata 22 11 BEC,:AK M.L. 1967, 1968
Hyla andersoni 12 DuELLMAN and CoLE 1965
Hyla arenicolor 12 DUELLMAN and COLE 1965
Hyla arborea 24 12 GALGAN01933
Hyla arborea japonica 24 12 IRIKI 1930
Hyla avivoca 24 12 BusHNELL et al. 1939
Hyla bistincta 12 DuELLMAN and CoLE 1965
Hyla cinerea 24 12 BusHNELL et al. 1939
Hyla crepitans 12 DuELLMAN and CoLE 1965
Hyla crepitans 24 12 present paper
Hyla crucifer crucifer 12 DuELLMAN and CoLE 1965
Hyla ebraccata 15 DuELLMAN and CoLE 1965
Hyla euphorbiacea 12 DUELLMAN and COLE 1965
Hyla faber 24 12 BEC,:AK M.L. 1967, 1968
Hyla fuscomarginata 24 12 present paper
Hyla hayi 24 12 BEc;:AK M.L. 1967, 1968
Hyla hazelae 24 12 DuELLMAN and CoLE 1965
Hyla lafrentzi 12 DuELLMAN and CoLE 1965
Hyla microcephala martini 15 DuELLMAN and CoLE 1965
Hyla microcephala microcephala 15 DuELLMAN and CoLE 1965
Hyla microps 30 15 BEc;:AK M.L. 1967, 1968
Hyla minuta 30 15 present paper
Hyla multilineata 24 12 8EC,:AK M.L. 1967, 1968
Hyla nana 30 15 present paper
Hyla polytaenia 24 12 present paper
H yla pulchella prasina 24 12 BEC,:AK M.L. 1967, 1968
Hyla pulchella pulchella 24 12 SAEZ and BRUM 1960
Hyla raniceps 30 15 present paper
 CHROMOSOMAL STUDIES IN ANURANS 49

(Table I cont.)
Species 2n n(~) Author

Hyla regilla 24 12 ~ORESCALCHI 1965

Hyla robertsorum 12 DuELLMAN and CoLE 1965
Hyla rubicundula 30 15 present paper
Hyla septentrionalis 12 DuELLMAN and CoLE 1965
Hyla smithi 12 DuELLMAN and CoLE 1965
Hyla trachytorax 24 12 present paper
Hyla versicolor 24 12 BusHNELL et al. 1939
Phyllomedusa burmeisteri 52 BE«;AK ~.L. et al. (In press)
Phyllomedusa calcarifer 13 DuELLMAN and CoLE 1965
Phyllomedusa callidryas callidryas 26 13 DuELLMAN and CoLE 1965
Phyllomedusa helenae 26 13 DuELLMAN and CoLE 1965
Phrynohyas spilomma 12 DUELLMAN and COLE 1965
Phrynohyas venulosa 24 12 present paper
Pseudacris brachyphona 24 12 DuELLMAN and CoLE 1965
Pseudacris triseriata kalmi 12 DuELLMAN and CoLE 1965
Pseudacris triseriata triseriata 24 12 DuELLMAN and CoLE 1965
Pternohyla fodiens 12 DuELLMAN and CoLE 1965
Ptychohyla ignicolor 12 DuELLMAN and CoLE 1965
Smilisca baudini 12 DuELLMAN and CoLE 1965
Smilisca cyanostica 12 DuELLMAN and CoLE 1965
Triprion petasatus 12 DuELLMAN and CoLE 1965

Family Centrolenidae
Centronella fleischmanni 10 DuELLMAN and CoLE 1965
Family Ranidae
Mantella aurantiaca 26 13 ~ORESCALCHI 1967
Rana agilis 26 SATO 1934
Rana arvalis 24 12 WICKBOM 1945
Rana arvalis 26 13 CEI 1946
Rana catesbeana 26 SWINGLE 1917
Rana catesbeana 28 14 SWINGLE 1921
Rana catesbeana 26 13 ~AKINO 1947
Rana dalmatina 26 13 CEI 1946
Rana esculenta 24 ScHOTTLANDER 1888
VON RATH 1895
Rana esculenta 16 CHAMPY 1913
Rana esculenta ca 25 13(~1) LEVY 1915
12,13 (~II)
Rana esculenta 26 13 GALGANO 1931; WICKBOM 1945;
CEI 1946
Rana graeca 26 13 CEI 1946
Rana iaponica 26 KAWAMURA 1939, 1943
Rana iaponica 26 u KoBAYASHI 1946
Rana limnocharis 26 13 SATO 1933, 1934
Rana nigromaculata 26 13 JRIKI 1928, 1932
Rana nigromaculata 13 TING 1936
Rana pipiens 2H 13 ~(~I) SWIN=LE 1917
Rana pipiens 26~ 12,13 ~(~II)
26 PARMENTER 1925
DIBERARDINO 1962
Rana plancyi 26 13 TING 1939
Rana rugosa 26 13 IRIKI 1928, 1932
Rana sp 16 DEKHUYZEN 1891
Rana sp 24 KAWAMURA 1943
KoBAYASHI 1946
26 1•3 WICKBOM 1945
GoLDSHMIDT 1920
50 RABELLO

(Table I cont.)
Species 2n n(~) Author

Rana tigrina 26 1•3 AsANA, apud 0GUMA and MAKINO

1937
Rana temporaria 8(MI) BETACHINI 1896
Rana temporaria 8-10~ (MI) CARNOY and LEBRUN 1900
Rana temporaria 10 ~(Mil) LEBRUN 1901
Rana temporaria 24 12 VON RATH 1895
Rana temporaria (R. fusca) 26 13 WITSCHI 1922, 1924
MAKINO 1932, 1946
Rana temporaria ornativentris 24 12 KoBAYASHI 1946
Family Phrynomeridae
Phrynomerus bifasciatus 26 MoRESCALCHI 1968
Family Rhacophoridae
Hyperolius argentivittis 26 MoRESCALCHI 1968
Kassina senegalensis 24 MoRESCALCHI 1968
Rhacophorus buergeri 26 SATO 1934
Rhacophorus schlegeli schlegeli 26 MAKINO 1932
Family Brachycephalidae
Melanophryniscus moreirae 22 11 BEc;AK M.L. et al. 1969
Melanophryniscus stelzneri
( =Atelopus stelzneri) 22 MoRESCALCHI and (;ARGIULO 1968
Family Microhylidae
Breviceps gibbosus 24 MORESCALCHI 1968
« Cacopoides tornieri » 28 14 SATO 1936
Dermatonotus miilleri 22 11 present paper
Gastrophryne carolinensis 22 MoRESCALCHI 1968
Kaloula pulchra 28 MORESCALCHI 1968

MATERIAL AND METHODS

The frogs included in this investigation were:

Family Hylidae:
Hyla raniceps (COPE 1862): five males and one female.
Phrynohyas venulosa (LAURENTIUS 1768): one male and two females.
Hyla crepitans WIEn 1824: eleven males and one female.
Hyla polytaenia WrEn 1870: three males.
Hyla minuta PETERS 1872: twelve males and one female.
Hyla rubicundula REINHARDT and LiiTKEN 1882: one male.
Hyla nana BouLENGER 1889: five males.
Hyla fuscomarginata A. LuTz 1925: six males.
Hyla trachytorax MuLLER and HELLNICH 1936: s1x males and one female.

Family Microhylidae:
Dermatonotus mulleri (BOETTGER 1885}: eight males and one female.
Squash preparations for cytological examination were made in accordance
with the procedure described by OHNO et al. ( 1964 ). Mitotical and meiotic pre-
 CHROMOSOMAL STIJDIES IN ANURANS 51

parations were obtained from fragments of spleen, gut and gonads of animals pre-
treated with a 1% solution of colchicine (0.1 ml per/g of weight).

RESULTS
In the species from which both sexes were examined, male and female
showed similar karyotypes. The spermatocytes I of eight species exhibited
ring bivalents with terminal chiasmata. In two species - Hyla rubicundula
and Hyla nana - interstitial chiasmata were observed. No bivalent sug-
gested by its morphology or behavior the presence of heteromorphic sex
~hromosomes.

Family Hylidae:
Hyla raniceps. Diploid number: 24; n= 12. Chromosomes: metacen-
trics and submetacentrics (Fig. 1 ).
Phrynohyas venulosa. Diploid number: 24; n= 12. Chromosomes: me-
tacentrics and submetacentrics (Figs. 4, 13, 14).
Hyla crepitans. Diploid number: 24; n = 12. With the exception of
pair 6 which is acrocentric all the other chromosomes are metacentric and
submetacentric (Fig. 5). Pair 6 has satellites on the short arms.
Hyla polytaenia. Diploid number: 24; n= 12. Chromosomes: metacen-
trics and submetacentrics (Fig. 3 ).
Hyla minuta. Diploid number: 30; n= 15. Chromosomes: metacentrics
and submetacentrics (Fig. 7).
Hyla rubicundula. Diploid number: 30; n= 15. Chromosomes: meta-
centrics and submetacentrics with the exception of pairs 7, 12, 13 and 15
which are acrocentric (Figs. 8, 15, 16). Interstitial chiasmata were observed
in the bivalents during metaphase I.
Hyla nana. Diploid number: 30; n= 15. Chromosomes: Pairs 5, 9, 14
and 15 are acrocentric; the others are metacentric and submetacentric
(Fig. 9). The bivalents in metaphase I exhibit interstitial chiasmata.
Hyla fuscomarginata. Diploid number: 24; n= 12. Chromosomes: me-
tacentrics and submetacentrics. Pairs 5 and 11 have satellites on the short
arms (Fig. 2).
Hyla trachytorax. Diploid number: 24; n = 12. Chromosomes: meta-
centrics and submetacentrics (Figs. 10, 11, 12). In the gonad of a male spe-
cimen, we have found 16,2% of mitosis with 25 chromosomes and 14,6%
of metaphases I with 12 bivalents and one univalent. The extra chromo-
some does not differ in size from the pairs 10, 11 and 12; it is not hetero-
picnotic and appears as a ring univalent in metaphase I.
Family Michrohylidae:
52 RABELLO

Dermatonotus miilleri. Diploid number: 22; n = 11. Chromosomes: me-

tacentrics and submetacentrics (Figs. 6, 17, 18 ).

Fig. 1. - Hyla raniceps, male karyotype (2n = 24).

Fig. 2. - Hyla fuscomarginata, male karyotype (2n = 24).
Fig. 3. - Hyla polytaenia, male karyotype (2n = 24).
Fig. 4. - Phrynohyas venulosa, male karyotype (2n = 24 ).
Fig. 5. - Hyla crepitans, male karyotype (2n = 24).
Fig. 6. - Dermatonotus miilleri, male karyotype (2n = 22).
 CHROMOSOMAL STUDIES IN ANURANS 53

Fig. 7. - Hyla minuta, male karyotype (2n = 30).

Fig. 8. - Hyla rubicundula, male karyotype (2n = 30).
Fig. 9. - Hyla nana, male karyotype (2n = 30).
Fig. 10. - Hyla trachytorax, male karyotype (2n = 24).
Fig. 11. - Hyla trachytorax, male karyotype with extra-chromosome (2n = 25).
Fig. 12. - Hyla trach)•torax, male diakinesis showing 12 bivalents and one ring univalent.

DISCUSSION

Diploid numbers range from 22 to 52 in Hylidae (Table 1).

According to WICKBOM (1945, 1949), the diversity of the diploid num-
bers among anurans can be attributed to centric fusions. M. L. BE~AK (1967,
1968) suggests that the available data favours the idea that the Robertsonian
mechanism acts at the level of related species within a given family, but
not between families. Pericentric inversions and translocations can make
this mechanism less evident. As shown in Table II, there is an obvious
54 RABELLO
 CHROMOSOMAL STUDIES IN ANURANS 55

assoc1at10n between high diploid numbers and numbers of acrocentrics m

the species for which data are available for this comparison (this paper
and those of M. L. BE<;AK 1967, 1968).
Cytological and cytophotometric studies (M. L. BE<;AK et al. 1966,
1967; W. BE<;AK et al. 1967) explained the variation in the diploid numbers
and DNA values in the Ceratophrydidae as a consequence of polyploidy.

TABLE II
Numbers and types of chromosomes in Hylidae.

Metacentric and Acrocentric

Species 2n submetacentric Author
pairs
pairs

Hyla albomarginata 22 11 M.L. BE(,:AK

H. hayi 24 12 M.L. BE(,:AK
H. pulchella prasina 24 12 M.L. BE(,:AK
H. multilineata 24 12 M.L. BE(,:AK
Phrynohyas venulosa 24 12 Present paper
H. polytaenia 24 12 )) ))

H. raniceps 24 12 )) ))

H. fuscomarginata 24 12 )) ))

H. trachytorax 24 12 )) ))

H. crepitans 24 11 1 )) ))

H. albomarginata 24 11 1 M. L. BE(,:AK
H. faber 24 10 2 M.L. BE(,:AK
Fritziana goeldii 26 10 3 M.L. BE(,:AK
H. minuta 30 15 Present paper
H. microps 30 11 4 M.L. BE(,:AK
H. rubicundula 30 11 4 Present paper

In fact, Odontophrynus americanus (4n = 44) and Ceratophrys dorsata

( 8n = 104) were the first known cases of bisexual polyploid species among
vertebrates. Recently, M. L. BE<;AK et al. (in press) reported polyploidy to
have occurred also in the family Hylidae, describing Phylomedusa burmeisteri
( 4n =52) as a tetraploid species.
Several mechanisms may influence diversification within a family. Alle-
lic mutations are, no doubt, important in early speciation. However, they

Fig. 13. - Phrynohyas venulosa, male metaphase I (12 bivalents).

Fig. 14. - Phrynohyas venulosa, male metaphase II (12 dyads).
Fig. 15. - Hyla rubicundula, male metaphase I ( 15 bivalents).
Fig. 16. - Hyla rubicundula, male metaphase II (15 dyads).
Fig. 17. - Dermatonotus mulleri, male metaphase I (11 bivalents)
Fig. 18. - Dermatonotus mulleri, male metaphase II (11 dyads).
56 RABELLO

may not explain by themselves all the diversification within a group. Gene
duplications are also fundamental, since redundance of the genetic material
is a pre-requisite to the formation of new loci (OHNO 1967). Gene products,
which were previously selected for, keep being provided by one of the du-
plicated loci, while the other becomes free for exploring further adaptive
possibilities through mutations, thus providing evolution with the necessary
variability. The species is enabled by repeated mutations to try new forms
of integration to the environment, without endangering its survival.
A recent survey of DNA values in the families Bufonidae, Brachycepha-
lidae, Hylidae, Microhylidae, Leptodactylidae and Ceratophrydidae revealed
the anurans to be an heterogenous group in this regard (W. BE~AK et al.
in press). DNA content ranges from 43% to 181% that of placental mam-
mals respectively in Leptodactylus ocellatus (2n=22) and Ceratophrys dor-
sata ( 8n = 104 ). Among Microhylidae, Dermatonotus mulleri ( 2n = 22) pre-
sents one of the lowest DNA values in anurans: 44% that of mammals.
Hylidae vary widely in their DNA contents. While Hyla nana (2n=30)
has a low DNA value (54%), H. pulchella prasina (2n=24) has 147%
that of mammals. Species with the same diploid number ( 2n = 24) such as
Hyla fuscomarginata, H. polytaenia, H. albomarginata, H. faber, H. multi-
lineata and H. pulchella prasina have different DNA values. According to
W. BE~AK et al. (in press), gene duplications accounted partially for the
differences of DNA content in the species they studied. Similar conclusions
were achieved by U LLERICH ( 196 7) in comparing species of Ran a and Bufo.
The presence in H. trachytorax of mitosis with 24 and 25 chromosomes
and of metaphases I with 12 bivalents side by side to others with 12 biva-
lents and one univalent could be due to the occurence of an early non-
disjunction in the gonads. Two cell lineages were formed: one with 23 chro-
mosomes, which would have been lethal, and another with 25. The last one,
developing paralell to the original lineage, would produce the mosaic. Yet,
the extra chromosome could be a supernumerary. ULLERICH (1967) repor-
ted the presence of supernumeraries in Rana temporaria. Such chromosomes
were heterochromatic and smaller than the smallest chromosome of the stan-
dard karyotype. In the case of H. trachytorax, the extra chromosome does
not differ in size from the pairs 10, 11 and 12 and it is not heteropicnotic.
The fact that it appears as a ring univalent in metaphase I does not necessa-
rily mean that it is not homologue to one of the pairs of the karyotype.
Since only a small sample of cells and animals was examined, this chromo-
some could, by chance, fail to form a trivalent in the cells studied. Only
a populational study may show which hypothesis is correct.
Our observations indicate the lack of heteromorphic sex chromosomes
in the species studied, in accordance with the observations of M. L. BE~AK
 CHROMOSOMAL STUDIES IN ANURANS 57

(1967, 1968) and of several authors in other anurans (STOHLER 1928; GAL-
GANO 1933, 1941; SAEZ et al. 1935). When both sexes were available, male
and female presented identic karyotypes.
The lack of morphological differentiation at the choromosomic level
does not imply, however, in an absence of genetical digamety. Sex determi-
nation in these anurans could be the task of one pair of chromosomes still
morphologically indifferentiated, status found by W. BE<;AK (1962, 1965,
1966) among the Boidae (Serpentes). An accumulation of heteromorphic sex
determining genes could be occuring in one chromosome by decreasing the
rate of crossing-over in a given pair, the morphological differentiation of
this chromosome having not yet begun.
Another hypothesis suggests that the sex in these amphibians is de-
termined by one or several pairs of male and female determining genes
distributed among several chromosomes.
Acknowledgements. - The author wishes to thank DR. M. L. BEc;:AK and DR. W. BE-
c;:AK for their helpful suggestions and kind revision of the manuscript; to W. BoKERMANN, J. }IM
and DR. L. D. VIZOTTO who supplied and classified the animals used in this investigation and
to M. KuCHENBUCH for typewriting.

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SUMMARY
The karyotypes and male meiOSIS of nine Hylidae and one Microhylidae are described.
Those are: Hylidae - Hyla raniceps (2n=24), Phrynohyas venulosa (2n=24), Hyla crepitans
(2n=24), Hyla polytaenia (2n=24), Hyla minuta (2n=30), Hyla rubicundula (2n=30), Hyla
nana (2n=30), Hyla fuscomarginata (2n=24), Hyla trachytorax (2n=24); Microhylidae-Derma-
tonotus miilleri (2n=22). Male and female karyotypes are identical. The spermatocytes I of eight
species exhibited ring bivalents with terminal chiasmata. Interstitial chiasmata were observed in
H. rubicundula and H. nana. No bivalent suggested by its morphology or behavior the presence
of heteromorphic sex chromosomes.
A male specimen of H. trachytorax (2n=24) showed in its gonads mitosis with 24 and
with 25 chromosomes and metaphases I both with 12 bivalents and with 12 bivalents and one
univalent.
The association between high diploid numbers and numbers of acrocentrics found in Hyli-
dae suggests that the mechanism of centric fusions occurred parallel to evolution in this family.