Corresponding author: Abel Batista (
Academic editor: J. Penner
Substantial molecular and morphological character differences lead us to the description of a new species of the genus
Mebert K, González-Pinzón M, Miranda M, Griffith E, Vesely M, Schmid PL, Batista A (2022) A new rainfrog of the genus
Tropical regions are extraordinarily rich in biodiversity which is caused by the combination of historical, climatic, and geographic characteristics that favor high speciation rates, as for example in anurans (
Currently, there are 13 species of
Our study focuses on Darién Province with the principal material originating from Cerro Chucantí. This mountain supports a remarkable diversity of organisms with 13 recently described new species, including plants (
Herein we describe a new species of
The primary study site is Cerro Chucantí that includes the highest peak (1439 m a.s.l. at
Map showing locations of
Additional material for molecular and/or morphological analysis was obtained from specimens collected in eastern Panama (location names in bold as used in article): 1)
For molecular analyses of Panamanian samples, DNA was extracted from fresh liver tissue. The 16S mtDNA extraction and sequencing follow previously described routines (
A Maximum Likelihood analysis (
Collecting permits for 2009 (SC/A-8-09, SC/A-28-09), 2011 (SC/A-37-11), 2012 (SC/A-33-12), 2016 (SE/A-60-16) and 2018 (SE/A-33-18) as well as export permits for 2012 (SC/A-33-12) and 2013 (SEX/A-7-13) were provided by UNARGEN-Ministerio de Ambiente, Panama. Finally, we applied the traditional Environmental Vulnerability Score (
Specimens were collected by hand, photographed alive, euthanized with the Solution Tanax T-61, fixed with a preservative solution of 5 mL formalin (36%) in 1 L ethanol (94%), and subsequently stored in ethanol (70%) following the protocol of
Sex of specimens was determined by morphometric characters and presence of eggs in Panamanian samples. Measurements were taken to the nearest 0.1 mm, using a stereomicroscope and precision digital calipers. Following variables were measured according to
Interspecific differences among
We conducted a Multiple Correspondence Analysis (
In the following, we present information on both genes (16S and COI) separately, as well as their combined results. However, we focus more on 16S for the presentation on closely related taxa, as 16S is more widely used and thus comparable with many Neotropical anurans (
The
Estimates of net evolutionary divergence (mean %) between groups (G-numbers from the
Species | 16S and COI evolutionary divergence between groups | ||||||||
---|---|---|---|---|---|---|---|---|---|
G1 | G2 | G3 | G4 | G5 | G6 | G7 | G8 | G9 | |
G1 |
– | – | – | – | – | – | – | – | – |
G2 |
9.6 | – | – | – | – | – | – | – | – |
G3 |
11.0 | 8.1 | – | – | – | – | – | – | – |
G4 |
13.7 | 12.7 | 14.6 | – | – | – | – | – | – |
G5 |
14.9 | 13.4 | 13.1 | 17.0 | – | – | – | – | – |
G6 |
14.6 | 12.8 | 14.3 | 13.2 | 17.5 | – | – | – | – |
G7 |
15.1 | 12.2 | 13.2 | 13.2 | 16.5 | 15.9 | – | – | – |
G8 |
16.4 | 12.7 | 14.2 | 15.7 | 18.0 | 16.0 | 18.3 | – | – |
G9 |
18.6 | 15.4 | 15.7 | 17.3 | 19.2 | 17.9 | 16.7 | 21.1 | – |
G10 |
22.0 | 20.6 | 20.5 | 22.4 | 22.7 | 22.0 | 20.6 | 22.3 | 21.7 |
Phylogenetic tree of
On a wider perspective, the phylogenetic inference based on combined 16S and COI sequences placed the
The shortest genetic distance for 16S mtDNA between the new
Results of morphometric measurements of adult specimens of
Morphometric characters of adult
Measurement | Females | Males |
---|---|---|
|
42.66±3.71(38.15–46.3; 4) | 31.24±3.52(26.9–36.7; 8) |
|
19.17±0.99(17.84–20; 4) | 12.39±1.55(10.7–15.9; 8) |
|
18.14±1.49(16.46–19.9; 4) | 12.44±1.69(10–14.7; 7) |
|
3.05±0.21(2.9–3.2; 2) | 2.37±0.55(1.67–3.3; 7) |
|
5.1±0.71(4.6–5.6; 2) | 3.11±0.45(2.6–3.9; 7) |
|
6.3±0.14(6.2–6.4; 2) | 4.79±0.43(4.3–5.4; 7) |
|
23.15±2.05(21.7–24.6; 2) | 14.61±5.07(3.5–18.8; 7) |
|
22.1±0.14(22–22.2; 2) | 14.45±1.99(11.7–17.8; 7) |
|
2.85±0.92(2.2–3.5; 2) | 1.34±0.49(0.65–2; 7) |
|
5.45±0.21(5.3–5.6; 2) | 4.11±0.61(3.7–5.3; 7) |
|
5.65±0.49(5.3–6; 2) | 3.55±0.67(3.02–5; 7) |
|
11.2±1.13(10.4–12; 2) | 7.73±0.85(6.5–8.86; 7) |
|
9.15±6.72(4.4–13.9; 2) | 9.22±1.27(7.5–11.4; 7) |
|
1.45±0.21(1.3–1.6; 2) | 0.72±0.28(0.41–1.1; 7) |
|
3.15±0.07(3.1–3.2; 2) | 1.66±0.26(1.2–2; 7) |
|
1.25±0.07(1.2–1.3; 2) | 0.59±0.28(0.19–0.94; 7) |
|
2.35±0.07(2.3–2.4; 2) | 1.24±0.35(0.66–1.7; 7) |
|
1.4±0(1.4–1.4; 2) | 0.62±0.16(0.39–0.83; 7) |
|
2.5±0.14(2.4–2.6; 2) | 1.39±0.11(1.3–1.6; 7) |
|
16.75±1.34(15.8–17.7; 2) | 8.46±1.2(7.14–10.8; 7) |
|
21.4±0.85(20.8–22; 2) | 12.85±1.73(10.5–14.8; 5) |
A
A subsequent Linear Discriminant Analysis
A final univariate analysis corroborates that in four morphometric variables used in the
A Multiple Correspondence Analysis (
The first and second dimension of the
Map of the Multiple Correspondence Analysis (
Two distinct clusters appear in the
U‑Test, W = 133, p < 0.001, and Dimension 2: Mann-Whitney U-Test, W = 321, p = 0.026). These results strongly separate the two
Based on molecular divergence and morphological consensus, we assign the undescribed
Coloration in life of specimens of
Seven males, three females. Male and female
This comparison includes only members of the
Two additional rainfrog taxa inhabit northwestern Colombia that are closely related to
Both,
Further detailed comparisons to similar rainfrog species, e.g.,
Preserved holotype of
(
(Fig.
(Fig.
The specific name is a noun in the genitive case and is a patronym in honor for Greta Thunberg, a Swedish student, and her global climate activism. Greta initiated a “School Strike for Climate Action” outside the Swedish parliament to demand a radical response to the threat by the ongoing climate change. Then sixteen-year-old Thunberg’s example has inspired students worldwide to carry out similar strikes called Fridays For Future that started in August 2018. In December 2018 she addressed world leaders at the COP24 climate talks in Katowice, Poland, with sharp and unmasked words, and equally impressed a global audience in January 2020 with her unpolitical, direct speech down to the point on “Averting a Climate Apocalypse” at the
Color pattern of specimens from Cerro Brewster, not included in the
Habitat, mating, and parental care in females of
Habitats occupied by
1 | Dorsal ground color uniform blackish or grayish, with white or orange blotches on groin, if not, the color is gray to pink, some species have an orange or yellow dorsolateral stripe, continuous or interrupted |
|
– | Dorsal ground color, cream, reddish, brown tones, green or olive, uniform or darker blotches or reticulations, without white or orange blotches on groin |
|
2 | White or orange blotches on groin |
|
– | Groin uniform |
|
3 | Well defined white blotches on groin |
|
– | Well defined orange blotches on groin |
|
4 | Dorsal ground color uniform gray to pink without dorsolateral stripes |
|
– | Orange or yellow dorsolateral stripes, continuous or interrupted, some specimens lack stripes, but dorsal color is blackish, never gray or pink |
|
5 | Dorsal ground color green or olive green, uniform or with darker blotches or reticulations |
|
– | Dorsal ground color, cream, reddish, brown, dark brown, or olive, uniform or with darker blotches or reticulations |
|
6 | Dorsal ground color uniform green, with or without a reddish brown transverse interorbital band, dorsal skin smooth |
|
– | Dorsal color green or olive, with irregular blotches or reticulations, brown, olive or reddish color, dorsal skin tuberculated |
|
7 | Heel smooth or with one to several similar small sized tubercles scattered over upper surface of hind limb; enlarged tubercle on upper eyelid present or not |
|
– | Well-developed pointed calcars, usually enlarged tubercle on upper eyelid |
|
8 | Presence of a dorsolateral granular folds, dorsal pattern with chevrons |
|
– | Dorsolateral region smooth, dorsal pattern uniform, never with chevron pattern |
|
9 | Anterior and posterior surfaces of thighs, calves, and feet red |
|
– | Uniform posterior surface of thigh |
|
10 | Posterior thighs uniform |
|
– | Posterior thighs dark brown with red-orange dots |
|
11 | Dorsal skin granulate or tuberculate, rarely smooth, head about as broad as long; snout rounded |
|
– | Dorsal skin smooth, long and pointed snout |
|
12 | No W-shape on dorsum, iris variable in color, usually highly reticulated or blackish |
|
– | W-shape ridge that extent from the back of the head to the shoulder region, groin, anterior, and posterior thigh red, iris usually pale golden without reticulation, eyes usually with an orange perimeter |
|
13 | Iris variably light colored, cream, yellow or reddish and strongly reticulated, venter heavily mottled with dark pigment to almost uniform black, upper surfaces gray, brown, brownish black; tympanic annulus partially evident in females, upper lips with dark patches, with light colored lips or parts of it little or not dark-bordered above |
|
– | Iris black, some very dark red, without reticulation, venter white, dirty white, yellow or red, upper surfaces reddish brown or yellow, white to yellow upper lips, contrastingly dark-bordered above, some with dark patches, tympanic annulus not visible in females |
|
The genus
Within Panama,
Whereas all interpopulation divergence between the type series with any of the other sites remains below 3%, other population comparisons can vary and increase up to 5.4% between single individuals from Cerro Brewster and El Cope, both central Panama. Similar minimum genetic differentiation of > 3–4% of the 16S rRNA gene have been found to associate to CCS and UCS (not yet described Confirmed and Unconfirmed Candidate Species) of frogs in Madagascar, Africa, and Amazon Basin, South America (
Cloud forests in general and isolated mountain tops in particular are highly vulnerable to climate change due they low range of mobility and high habitat specialization of its denizen (
We thank Luis DeLeon, Jesús Pérez, Juan Zarzavilla, Yorlis Cáceres, Hugo Martínez, and Gilberto Torres for field assistance, and Guido Berguido for his support during our stay at the Chucantí private reserve. We also thank to Sarah Farinelli, Juan Daza, Mauricio Rivera-Correa, and an anonymous reviewer for their valuable comments during the review process, which greatly improved the manuscript. To Marcos Ponce and Angel Sosa a thank you for providing photos of a specimen from Cerro Brewster. This work was financially supported by Asociación ADOPTA el Bosque Panama, Sistema Nacional de Investigación (SNI) of the Secretaría Nacional de Ciencia, Tecnología e Innovación (SENACYT, Panamá), and the Rainforest Trust (US). We thank Sebastian Lotzkat, for his assistance in the field and on the manuscript. Many colleagues have provided photographs for the species panels in the Suppl. material
Tables S1–S5
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A new rainfrog of the genus
Figures S1–S16
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A new rainfrog of the genus