Research Article |
Corresponding author: Bianca V.M. Berneck ( bvmberneck@gmail.com ) Academic editor: Anthony Herrel
© 2017 Bianca V.M. Berneck, Ariovaldo A. Giaretta, Reuber A. Brandão, Carlos A. G. Cruz, Celio F.B. Haddad.
This is an open access article distributed under the terms of the Creative Commons Attribution License (CC BY 4.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.
Citation:
Berneck BVM, Giaretta AA, Brandão RA, Cruz CAG, Haddad CFB (2017) The first species of Aplastodiscus endemic to the Brazilian Cerrado (Anura, Hylidae). ZooKeys 642: 115-130. https://doi.org/10.3897/zookeys.642.10401
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The genus Aplastodiscus includes 14 nominal species in four monophyletic groups with occurrence in the Atlantic Forest and Brazilian Cerrado (Brazilian Savanna) of South America. A recent study reviewed the taxonomy and phylogenetic relationships of the genus and suggested a third species for the A. perviridis Group. Herein, on the basis of morphology and advertisement call, we describe this species and test its monophyly. The new species is the only Aplastodiscus with endemic occurrence in the Cerrado Biome. In addition, its geographical distribution and conservation status are discussed.
Aplastodiscus lutzorum sp. n., Cophomantinae , new species, integrative taxonomy
The genus Aplastodiscus includes 14 nominal species in four monophyletic groups (
The following measurements follow
SVL snout-vent length
HL head length
HW head width
ED eye diameter
TD tympanum diameter
END eye-nostril distance
IOD interocular distance
THL thigh length
FL foot length; including tarsus
However, the tibia length (TBL) follows
The adult specimens are housed in the following Brazilian collections: Célio F. B. Haddad collection (CFBH) at the Universidade Estadual Paulista, Rio Claro, SP; Coleção Herpetológica da Universidade de Brasília (CHUNB) at the Universidade de Brasília, Brasília, DF; Museu de Ciências Naturais da Pontifícia Universidade Católica de Minas Gerais (MCN-AM), Belo Horizonte, MG; and Coleção de Anuros da Universidade Federal de Uberlândia (AAG-UFU), Uberlândia, MG.
Males of the new species were recorded in the Municipality of Alto Paraíso de Goiás, Goiás State (N = 6) and Brasília, Distrito Federal. For comparative purposes, males of A. perviridis were recorded at the type-locality (N = 5), in Serra da Bocaina, São José do Barreiro, São Paulo State. Calls were recorded with a Marantz PMD 671, a Boss BR-864 (both with a Sennheiser ME67/K6 microphone) or a MicroTrack (ME66/K6 microphone), all set at 44.1 kHz and 16-bit resolution. Calls were recorded from 50 to 150 cm from calling males, and 10 to 15 calls were analyzed for each male. Acoustic variables were analyzed with RAVEN PRO 1.5, 64-bit version (
Acoustic variables of the advertisement call of topotypes Aplastodiscus perviridis and A. lutzorum sp. n. n = number of recorded males.
Call Features |
A. lutzorum sp. n. (n = 12) Range Mean (SD) |
A. perviridis (n = 5) Range Mean (SD) |
||
---|---|---|---|---|
Call Duration (seconds) | 0.26–0.40 | 0.32 (0.05) | 0.12–0.15 | 0.13 (0.01) |
Higher Frequency (kHz) | 2334–2647 | 2468 (97) | 2419–2750 | 2519 (135) |
Lower Frequency (kHz) | 1494–1732 | 1595 (76) | 1587–1806 | 1690 (82) |
Dominant Peak (kHz) | 1884–2156 | 2027 (79) | 1981–2153 | 2078 (66) |
Time to Frequency Peak (%) | 49–70 | 61 (7) | 23–38 | 34 (6) |
Air temperature range | 19–22 °C | 16–19 °C | ||
Record hour | 20:00–22:00 h | 20:30–21:00 h |
The extraction of DNA was carried out using ethanol-preserved tissues and the DNeasy isolation kit (Qiagen, Valencia, CA, USA). We carried out DNA amplification in a 25 µL volume reaction using master mix Fermentas Taq Polymerase and reagents. The Polymerase chain reactions (PCR) included an initial denaturing step of 30s at 94 °C, followed by 35 cycles of amplification with a final extension step at 72 °C for 6 min. The products of PCR were sent for sequencing to Macrogen, South Korea. We sequenced DNA fragments in both directions to minimize potential errors. The chromatograms were read and edited using SEQUENCHER 3.0 (Gene Codes, Ann Arbor, MI, USA) and complete sequences were edited with MEGA 6.06 (
Sequence alignments were performed in Clustawl (
Aplastodiscus
perviridis
Aplastodiscus
perviridis
Aplastodiscus
perviridis
Aplastodiscus
perviridis
Aplastodiscus
sp.
Aplastodiscus
sp. 3
(Figs
CFBH 22777–80, four adult males collected at Fazenda Água Limpa, Brasília, Distrito Federal (15°56'55.45"S; 47°56'17.83"W) on 18 February 2009. AAG-UFU 863, 865-867 and AAG-UFU 1639 collected on December 2012, AAG-UFU 3343 on 29 November 2013, AAG-UFU 3350–51, 5073–76, 5091, AAG-UFU 0867, adult female, all collected with the holotype. CHUNB 17015–17016 adult males collected on 31 December 1995 at Alexânia, Goiás (16°5'42.00"S; 48°31'20.60"W), CHUNB 17018, adult male collected on 12 December 1985 at Alexânia, Goiás, and CHUNB 74504–74508 adult males from Fazenda Água Limpa, Brasília, Distrito Federal, collected on March 2013. All localities are in Brazil.
All males. MCN-AM 8809–12 and 8767–72 from AHE Queimado, Unaí, Minas Gerais (16°20'55.51"S; 46°52'48.93"W), collected on February–March 2007.
Aplastodiscus lutzorum sp. n. belongs to the A. perviridis Group and thus bears bicolored irises, lacks webbing between toes I and II, has reduced webbing among the remaining toes, and lacks peri-cloacal ornamentation. The new species is diagnosed by its small SVL for the A. perviridis Group (30–36 mm) and by the advertisement call 2.5 times longer.
Adult male: head 20% wider than long; snout rounded in profile, nearly rounded in dorsal view; canthus rostralis curved; loreal region concave; nostrils ovoid; internarial region grooved. Supratympanic fold distinct, from posterior corner of orbit to insertion of forearm; tympanum distinct, almost circular; tympanum diameter 48.5% of eye diameter. Upper eyelid smooth as the dorsum. Thoracic fold just discernible. External vocal sac single, subgular, and expanded. Fingers long, slender, no lateral fringe, bearing discret round terminal discs; relative lengths of fingers I, II, IV, III; similar sized discs on fingers II, III and IV, larger than disc of Finger I. Diameter of disc of Finger III equals to diameter of Toe IV, about 42% eye diameter. Subarticular tubercles well defined, rounded; supernumerary tubercles poorly defined on palm, rounded; inner metacarpal tubercle large, rounded, about four times the size of subarticular tubercles; other metacarpal tubercle barely defined; spine of prepollex absent; no nuptial pads; ulnar crest barely defined. Hand webbing formulae: I-II 2--3- III 2+-2 IV. Tarsal texture smooth; tarsal fold discrete, extending to the entire length of tarsus; heel smooth; inner metatarsal tubercles large, elongate, three times the size of foot subarticular tubercles; outer metatarsal tubercle absent; subarticular tubercles distinct, large, and rounded, diameter about 3/4 width of terminal disc on the same toe; supernumerary tubercles absent; toes long, slender, without lateral fringe; toes bearing rounded discs, smaller in diameter to those of fingers II-IV. Foot webbing formula: I 2+ - 3- II 2+ - 31/2 III 2+ - 4- IV 3+ - 2V. Supra cloacal fold absent. Skin on dorsum smooth; skin on throat, belly, ventral surface of thigh, and arm granular. Dorsal and dorsolateral surfaces almost entirely pale yellow with small dark spots or mottles on dorsal surfaces. Belly pale yellow. Measurements of the holotype (mm): SVL 34.6, HL 10.6, HW 11.4, ED 3.3, TD 2.1, END 3.2, IOD 5.3, THL 18.1, TBL 15.8, and FL 18.4 (Table
Measurements (in millimeters) of the type-series of Aplastodiscus lutzorum sp. n. Abbreviations are: SVL (snout-vent length), HL (head length), HW (head width), ED (eye diameter), TD (tympanum diameter), END (eye-nostril distance), IOD (interocular distance), THL (thigh length), TBL (tibia length), and FL (foot length).
Measurement | Holotype | Female paratype | Males paratypes N = 25 (Mean) |
---|---|---|---|
SVL | 34.6 | 33.7 | 30.7–36 (33.5) |
HL | 10.6 | 11.4 | 8.8–11.4 (10.5) |
HW | 11.4 | 11.1 | 10.5–12.4 (11.4) |
ED | 3.3 | 3.4 | 3–3.7 (3.4) |
TD | 2.1 | 2.4 | 1.5–2.4 (2) |
END | 3.2 | 3.1 | 1.6–3.3 (2.7) |
IOD | 5.3 | 5.7 | 4.5–5.9 (5.4) |
THL | 18.1 | 16.2 | 12–18.7 (17) |
TBL | 15.8 | 16.4 | 14.2–18.5 (16) |
FL | 18.3 | 18.8 | 14.9–19.6 (17) |
Color in life of the type-series: Dorsal head surface dark green, almost olive. Dorsal body surface and flanks yellowish green with small and scattered melanophores. The superior third of eye is golden, whereas the inferior 2/3 is red copper. Eye surrounded by a black ring. Vocal sac bluish green. In preservative, colors fade to pale beige and the dorsum shows several dark brown dots, making it darker than other parts of the body. The belly is uniformly pale yellow.
The main variation in type series is the body size (Table
Advertisement calls of A. lutzorum sp. n. (Figure
Above, audiospectrogram and oscillogram of three advertisement calls of the holotype of Aplastodiscus lutzorum sp. n. (Chapada dos Veadeiros, 12 December 2011, air temperature 20 °C); the background calls are from another male calling in antiphony. Bellow, audiospectrogram and oscillogram of three advertisement call of A. perviridis (Serra da Bocaina, 10, January 2012, air temperature 16 °C).
Aplastodiscus perviridis’ advertisement call (Figure
All specimens of Aplastodiscus lutzorum sp. n. were collected along gallery forests with scattered buriti (Mauritia flexuosa) palm trees within the Cerrado Biome (see also
Geographic distribution of Aplastodiscus cochranae (pink triangles), A. perviridis (green circle), and A. lutzorum sp. n. (blue squares, blue star indicates its type-locality). Note that A. lutzorum shows a disjunctive distribution regarding the other Aplastodiscus species, occurring deep within Cerrado Biome.
The new species is named after the Brazilian scientists Adolfo and Bertha Lutz, who were pioneers in discovering and studying Aplastodiscus and some species of Hyla now included in the genus Aplastodiscus.
Aplastodiscus lutzorum sp. n. can be distinguished from the species of other groups of Aplastodiscus (A. albosignatus, A. albofrenatus, and A. sibilatus groups) by the presence of bicolored irises, the lack of the webbing between toes I and II, the webbing among the remaining toes reduced, and the absence of peri-cloacal ornamentation. The new species is diagnosed from A. perviridis and A. cochranae by having smaller SVL (30–36 mm SVL in the new species, 38–44.7 mm in A. perviridis, and 41–46.4 mm in A. cochranae) and longer advertisement calls (0.38–0.42 in new species, 0.14–0.17 in A. perviridis and 0.10–0.18 in A. cochranae). From A. cochranae it can be also distinguished by the green dorsal color in life (A. cochranae is brown) and by the absence of a brown line on the loreal region and a white line in the cloacal region (both present in A. cochranae). (Figures
The two DNA fragments aligned and concatenated resulted in a matrix of 1,227pb. Our analysis recovered four most parsimonious trees with 808 steps each (Figure
Uncorrected pairwise sequence distances (p-distances) of the Cytochrome c oxidase subunit 1 (above the diagonal) and 16s fragments (below the diagonal) for the species of the Aplastodiscus perviridis species Group. See the Material and methods section for the primers that delimitate each fragment. Values are in percentage.
1 | 2 | 3 | 4 | 5 | 6 | 7 | 8 | 9 | 10 | 11 | 12 | ||
---|---|---|---|---|---|---|---|---|---|---|---|---|---|
1 | A. cochranae CFBH3001 Rancho Queimado, SC | — | 1.06 | 9.13 | – | – | 9.55 | 8.92 | 9.77 | 9.98 | 9.77 | 8.92 | 10.19 |
2 | A. cochranae CFBHT14968 Lauro Muller, SC | – | — | 9.34 | – | – | 9.77 | 9.34 | 9.98 | 10.19 | 9.98 | 9.13 | 10.4 |
3 | A. lutzorum sp. n. CFBH22778 Brasília, DF | 4.28 | – | — | – | – | 0.42 | 0.42 | 10.4 | 9.34 | 9.98 | 9.13 | 9.55 |
4 | A. lutzorum sp. n. AAG1316 Brasília, DF | 4.28 | – | 0.56 | — | – | – | – | – | – | – | – | – |
5 | A. lutzorum sp. n. AAG1317 Brasília, DF | 4.09 | – | 0.37 | 0.19 | — | – | – | – | – | – | – | – |
6 | A. lutzorum sp. n. AAG1333 Alto Paraíso, GO | 4.09 | – | 0.37 | 0.19 | 0 | — | 0.85 | 10.19 | 9.13 | 9.77 | 8.92 | 9.55 |
7 | A. lutzorum sp. n. AAG741 Alto Paraíso, GO | 4.46 | – | 0.93 | 0.19 | 0.56 | 0.56 | — | 10.4 | 9.34 | 9.98 | 9.13 | 9.55 |
8 | A. perviridis CFBH18119 Topotype | 3.35 | – | 5.2 | 5.2 | 5.02 | 5.02 | 5.58 | — | 5.3 | 6.37 | 4.03 | 5.1 |
9 | A. perviridis CFBH7195 Santo Antônio do Pinhal, SP | 3.16 | – | 5.39 | 5.39 | 5.2 | 5.2 | 5.76 | 2.23 | — | 6.16 | 4.25 | 0.64 |
10 | A. perviridis CFBHT270 São Bento do Sul, SC | 2.23 | – | 4.83 | 4.83 | 4.65 | 4.65 | 5.02 | 2.79 | 2.42 | — | 2.76 | 5.94 |
11 | A. perviridis MACN37791 Misiones, Argentina | 3.16 | – | 5.76 | 5.76 | 5.58 | 5.58 | 5.95 | 3.35 | 3.16 | 0.93 | — | 4.03 |
12 | A. perviridis AAG1259 Atibaia, SP | 2.97 | – | 5.39 | 5.39 | 5.2 | 5.2 | 5.76 | 2.23 | 0.37 | 2.42 | 2.97 | — |
The A. perviridis Group now includes a third species, Aplastodiscus lutzorum, a species diagnosed mainly by its advertisement call, small size, and genetic differentiation. Genetic p-distances and phylogenetic topology support our hypothesis of the new species.
The scope of this paper was not to test biogeographic hypotheses. However, Aplastodiscus lutzorum is the only species of Aplastodiscus that occurs deep in the Cerrado Biome (see
A population from the dam of Queimado in the municipality of Unaí, in the state of Minas Gerais, Brazil (an area flooded by the construction of a hydroelectric station) was included as “referred specimens” for A. lutzorum The conservation status of this population is unknown. We consider the new species to be listed as a “Least Concern”, due to the fact that most of its area of occurrence is in protected places, such as the Parque Nacional da Chapada dos Veadeiros, Área de Relevante Interesse Ecológico (ARIE) do Capetinga/Taquara (Fazenda Água Limpa), Estação Ecológica de Águas Emendadas, and Floresta Nacional de Silvânia.
We are grateful to Luciana B. Nascimento (MCN-AM) and Guarino R. Colli (CHUNB) for allowing access to some specimens included in this work; and to the Centro de Estudos de Insetos Sociais (CEIS), UNESP, Rio Claro, Brazil, for allowing the use of their facilities for molecular analysis. For licenses, we thank the Instituto Chico Mendes (IBAMA, #17168). For assistance in fieldwork, we thank M. Segalla, P. Valdujo, D. Velho, K. G. Facure and C. S. Bernarde. V. G. D. Orrico made helpful comments on a draft. We thank to John Karpinsk for English review. The editor A. Herrel and reviewers Helio da Silva and Ivan Nunes improved this manuscript. BVMB would like to thank: J. Faivovich for suggestions and support, D. Baêta for discussions on taxonomy, the CFBH staff from 2008–16, and the São Paulo Research Foundation (FAPESP) for grants #2008/55235-4, #2013/18807-8, and #2013/50741-7. Financial support was provided to AAG by CNPq and FAPEMIG; a grant by CNPq. AAG collection permits: IBAMA 29954–3 and IBAMA 02015.008064/02–51. Financial support to CAGC and CFBH by CNPq.
Analyzed sound files; from Ariovaldo A. Giaretta collection
Data type: species data
List of voucher specimens, GenBank accession numbers, and locality data
Data type: species data