A KARYOTYPIC REVIEW OF 47 I II III

SOUTH AMERICAN BUFONIDS

Centrómeros DAPi(+) y Banda DAPI(-) en el cromosoma portador de la NOR 7p

(ANURA, BUFONIDAE) M. alipioi. Osornophryne sp. Rhinella ornata.

Martínez MN①, Ferro JM①, Cotichelli L②, Taffarel A①, Blasco-Zúniga A③, Pereyra M④, Borschein R⑤, Pie M⑥, Borteiro C⑦, Centromeric DAPI(+) marks Centromeric DAPI(-) marks Without marks besides the DAPI(-)
(One chromosome is missing) marks on NORs
Kolenc F⑦, Rivera M③, Baldo D①
IV
INTRODUCTION MATERIALS & METHODS
Cytogenetic information for the Bufonidae family is scarce, with data for only 130 of Mitotic preparations were obtained from intestines
the 609 species making up the family (Frost, 2018), where in most cases it is limited and bone marrow following Schmid et al. (2010). N. variegata. Interstitial Dapi(+) marks on both arms of each chromosome.

to reports of the chromosome number. The main goal of this work is to present a Techniques:
Figure ❷: Representative patterns of DAPI fluorochrome staining on studied
Ÿ C-Bands (Sumner, 1972)
cytogenetic review of 47 species of bufonids that belong to the neotropical genera Ÿ DAPI fluorochrome staining (Schweizer, 1976).
species. On Figure 1 it is indicated the patterns for eachgenus or species group,
Melanophryniscus (19 spp.), Dendrophryniscus (1 sp.), Osornophryne (1 sp.), denoted with Roman numerals. Arrows show DAPI(-) marks coincident with
Ÿ Ag-NOR (Howell & Black, 1980).
secondary constriction and Ag-NOR.
Atelopus (1 sp.), Amazophrynella (2 spp.), Nannophryne (1 sp.), Rhaebo (1 sp.), and 2n: somatic number
Rhinella (21 spp.). We interpret our results in a phylogenetic framework and discuss X: chromosome number
about the patterns of evolution observed in some chromosomal characters. NF: fundamental number RESULTS AND DISCUSSION
FIGURE ❶: Representative C-bands, Ag-NORs and DAPI fluorochrome staining patterns for the genus and species group studied, mapped on a resumed n = x= : synapomorphy for the Conmutabirana clade
phylogenetic hypothesis for Bufonidae (Portik and Pappenfus, 2015). Arrowheads show interstitial and telomeric C-bands. Asterisks indicate chromosome pairs (Bufonidae, Alloprhynidae, Centrolenidae, Odontophrynidae,
where only one of them marks positive for the NOR with silver staining. The patterns observed for DAPI are the ones shown on the Figure 2, and denoted with
Roman numerals.
Leptodactylidae and Brachycephaloidea) proposed by Streicher et al.
(2017).
CB NOR DAPI
general pattern
FN= : except for Osornophryne sp. (FN= ) and Amazophrynella
bokermanni (FN= ).
M. stelzneri group
Representative karyotypes of C-Band, chromosomes bearing the Ag-NOR and
M. rubriventris 8
I corresponding patterns of DAPI fluorochrome staining for each genus are presented on
figure 2, mapped on a resumed phylogenetic hypothesis for Bufonidae obtained from
Melanophryniscus
M. tumifrons group the analysis of Portik and Pappenfus (2015).

M. pachyrhynus 5
I C-B :
Karyotypes showed heterochromatin mostly restricted to the
M. moreirae group
centromeric or pericentromeric chromosome regions, although some
I
M. xanthostomus 7* interstitial and telomeric bands were also observed (arrowheads on
Dendrophryniscus
Figure 1 and Figure 3).
We could not get good quality C-Bands for Dendrophryniscus
III
D. berthalutzae
(conventional staining)
1* 10* berthalutzae.
Atelopus S S
III
Atelopus sp. 8 Ag-NOR on pair 8: putative plesiomorphic state for
Osornophryne
Bufonidae, although it is necessary to include M. moreirae in a
II
phylogenetic study to confirm this hypothesis.
Osornophryne sp. 6
Ag- NOR on pair 7: putative synapomorphy for a less
inclusive clade within Bufonidae, comprising Amazophrynella
Amazophrynella
- and all the other derived bufonids.
A. bokermanni 4*
Other states of this character would represent apomorphies at different levels, but this
interpretation is limited by the lack of information for some taxa.
III
A. minuta 7 DAPI F S
Nannophryne
There are four different patterns for DAPI stainning (wich positive and
negative marks indicate AT or GC rich heterochromatin blocks
IV
N. variegata 7 respectively), shown on Figure 2. On Figure 1 we indicate with Roman
Rhaebo
numerals the patterns for genus or species group.
- In all cases the NOR position is coincident with DAPI negative marks.
R. guttatus 7
Other positive and negative marks, coincident with C-bands are
R. spinulosa
group
shown on Figure 3.
I
R. spinulosa 11 CONCLUSION Figure ❸: Other hetero-
chromatin blocks different
R. veraguensis The present work brings up novel from the ones evidenced by C-
group Banding an DAPI staining.
III information about cytogenetics characters CB DAPI
R. rumbolli 11
Rhinella for Bufonidae. We studied for the first time
R. margaritfera
group
the karyotypes of 21 species ( spp. from 4 4
- Rhinalla, spp. from Melaphryniscus, sp. M. pachyrrinus
R. margaritifera 7

R. granulosa from Atelopus, sp. from Osornophryne, sp.

group 5 6 5 6
from Dendrophryniscus and 2spp. from M. sanmartini (M. moreirae group)

III Amazophrynella) and redescribed the

R. mirandariberoi 5
R. crucifer karyotypes of 27 species (9spp. from 3
Rhinella aff spinulosa
3
group
Rhinella, 15spp. from Melanophryniscus, 1sp.
III
R. ornata 7 from Nannophryne and 1sp. from Rhaebo) 1 3 1 3
R. marina
group
with new techniques. Rhinella aff spinulosa

❶Laboratorio de Genética Evolutiva, Instituto de Biología Subtropical (CONICET-UNaM), Facultad de Ciencias Exactas Químicas y Naturales, Universidad Nacional de
Misiones. Posadas, Misiones, Argentina.
I & III ❷Laboratorio de Sistemática y Biología de Anfibios, Instituto de Diversidad y Evolución Austral (IDEAus-CONICET), Puerto Madryn, Chubut, Argentina.
R. achavali 7 ❸Museo de Zoología de la Pontificia Universidad Católica del Ecuador, Quito, Ecuador.
❹Museo Argentino de Ciencias Naturales "Bernardino Rivadavia"-CONICET. Av. Ángel Gallardo 470. Buenos Aires, Argentina.
❺Instituto de Ciencias Biologicas, Universidade Federal de Minas Gerais, Belo Horizonte, Minas Gerais, Brasil.
❻Departamento de Zoologia, Universidade Federal do Paraná, Curitiba, Paraná, Brasil.
❼Sección Herpetología, Museo Nacional de Historia Natural, 25 de mayo 582, Montevideo, Uruguay.