Pristimantis conservatio

Barrio-Amorós, César L., Heinicke, Matthew P. & Hedges, S. Blair, 2013, A new tuberculated Pristimantis (Anura, Terrarana, Strabomantidae) from the Venezuelan Andes, redescription of Pristimantis pleurostriatus, and variation within Pristimantis vanadisae, Zootaxa 3647 (1), pp. 43-62 : 57-62

Relationships of P. conservatio and other Venezuelan Pristimantis within Terrarana

There are no known synapomorphies for Pristimantis . However, Hedges et al. (2008) list the following combination of features as typical for Pristimantis : head as wide as body, tympanic membrane differentiated, dentigerous process of vomers present, terminal discs of digits expanded, bearing well defined circumferential grooves, toe V usually longer than toe III. Pristimantis conservatio conforms to these characteristics, and molecular phylogenetic analyses additionally place this species within Pristimantis ( Fig. 8 ). We assign tentatively P. conservatio to the P. unistrigatus species group as defined by Lynch & Duellman (1997) and Hedges et al. (2008) by having slender bodies, being small to medium sized frogs, with short snouts and moderately long limbs, having finger I shorter than finger II, toe V longer than toe III, the digital discs expanded, the tympanic annulus and membrane present, without cranial crests, vomerine teeth and vocal sacs present.

We caution, however, that most of the classical species groups of Pristimantis based on morphology (Lynch & Duellman 1997) have yet to be supported in molecular phylogenies (Hedges et al. 2008; Pinto-Sánchez et al. 2012). Pinto-Sánchez et al. (2012) attributed, incorrectly, those morphologically-based species groups to Hedges et al. (2008). Those latter authors only listed the groups defined by earlier authors (Lynch & Duellman 1997), for convenience, pointing out that "none of these phenetic groups has been clearly distinguished in various phylogenetic analyses" (Hedges et al. 2008). Pinto-Sánchez et al. (2012) concluded that "the subgeneric taxonomy of Pristimantis is clearly flawed," but this was based largely on their misunderstandings of the earlier work. Neither those authors, or Hedges et al. (2008), sampled taxa sufficiently for a reclassification of Pristimantis , and therefore claims of rejection or support of supraspecific taxa (Pinto-Sánchez et al. 2012) are premature.

Likelihood and parsimony analyses recovered highly concordant trees ( Fig. 8 ). Pristimantis conservatio is confirmed as a member of Pristimantis , with monophyly of the genus receiving significant support. The species is also distinct from P. vanadisae , with uncorrected pairwise genetic distances of 5%–6%, vs. 0%–2% within P. vanadisae . These distances are somewhat smaller than distances between Pristimantis species for more rapidlyevolving mitochondrial genes like ND2 or cytochrome b (Elmer et al. 2007, Wang et al. 2008), but are consistent with species-level differences in homologous 12S and 16S gene regions observed among sets of related species of Pristimantis (e.g. Padial & De la Riva, 2009). For example, inspection of a dataset of 12S and 16S sequences included in Hedges et al. (2008) reveals 47 Pristimantis species pairs having uncorrected pairwise 12S/16S genetic distances at or below 5% (out of 5460 possible pairwise comparisons; the dataset included 105 Pristimantis species).

The series of individuals representing P. vanadisae is monophyletic, and includes individuals exhibiting pattern forms A, C, and D, confirming the presence of pattern polymorphism in P. vanadisae . CVULA 7184 from Altos de San Luis, was identified as P. aff. vanadisae , due to its similar morphology, but in our phylogeny is sister to P. conservatio , with a pairwise genetic distance of 3.4% from P. conservatio , vs. 5.0% from P. vanadisae . This point to another similar species still to be described. Additional Venezuelan taxa recovered as nested within Pristimantis include P. ginesi (Rivero, 1964) , P. b r i c e n i (Boulenger, 1903), P. lancinii (Donoso-Barros, 1968) , P. paramerus (Rivero, 1982) , and P. pleurostriatus . Eupsophus ginesi (= Pristimantis ginesi ) is the type species of Paramophrynella La Marca, 2007 , supporting placement of this genus in the synonymy of Pristimantis , following the nomenclature of Hedges et al. (2008). Pristimantis briceni , P. lancinii and P. paramerus were placed in Mucubatrachus by La Marca (2007), with Hylodes briceni (= Pristimantis briceni ) designated as type species. While these three species plus an undescribed species form a monophyletic grouping in our phylogeny, it is clear that this genus also is embedded within Pristimantis sensu Hedges et al. (2008) . As recognition of either genus at this time would render Pristimantis paraphyletic, we advocate maintaining both Mucubatrachus and Paramophrynella in the synonymy of Pristimantis at present. Recognition of one or both at the genus or subgenus level may be warranted in the future considering the current unwieldy nature of Pristimantis , with well over 400 recognized species (Frost 2013). However, such action should only be taken in the context of a broadly-sampled revisionary work.

Discussion

In this work we attempt to clarify the taxonomy of the tuberculated Pristimantis from the Cordillera de Mérida (Andean Venezuela). Until now, only one species was known to have ulnar, tarsal and conical heel tubercles: P. vanadisae . This species is distributed throughout the central Cordillera de Mérida, both at Sierra Nevada and Sierra de la Culata, and the name P. vanadisae has been associated with most specimens encountered with such tubercles in Venezuelan collections (e.g. CVULA). The reality is more complex, as P. vanadisae is polychromatic, varying within and among populations, while P. pleurostriatus and P. conservatio sp. nov. also are characterized by such tubercles. Rivero (1984 1982, Rivero & Segu 1989) also commented on the apparent disproportionate sex ratio in favor of the females of Eleutherodactylus cerasoventris (= P. vanadisae ), of which he never saw a male. This was taken by Rivero (Rivero & Segu 1989) as a suggestion that P. vanadisae could be a parthenogenetic species, as he dissected 104 specimens and all proved to be females. La Marca (1984) in the original description of P. vanadisae , mention three males vs. 26 females, additional evidence of biased sex ratio. Even if males are extremely rare, the fact that no call can be associated with P. vanadisae is apparently true; the senior author never has associated a call with any known population of P. vanadisae (La Macana, La Bravera, Monte Zerpa, La Mucuy) . Our experience is different with P. pleurostriatus , of which the majority of the animals known are males (nine males, one female); and almost all were calling when captured.

Pristimantis pleurostriatus is not a common species, and its limited area of distribution suggests that it be categorized as VU B2ab(i,ii) under the IUCN criteria (Stuart et al. 2008). On the other hand, P. conservatio may be found in similar areas of cloud forest in the eastern versant of the Cordillera de Mérida, as there are no significant geographic barriers to suggest that its distribution should be so limited. We suggest classifying it as DD (data deficient).

Acknowledgements

We are deeply thankful to Jorge Bravo, owner of Estancia La Bravera, for letting us to use his place as base camp for expeditions to La Carbonera and surroundings as a part of the agreement between Fundación AndígenA and Estancia La Bravera, permitting this way the redescription of Pristimantis pleurostriatus . Partial funding was provided through Project “Estatus de la población de la rana infectada por hongo Aromobates meridensis ( Anura : Aromobatidae ), en los Andes de Venezuela from IEA-Venezuela. John Jairo Mueses-Cisneros and Diego Cisneros- Heredia responded to questions about the diversity of Pristimantis in their respective countries ( Colombia and Ecuador). Javier García and Ursula Vera were great field companions. The animals were collected under a permit from Ministerio del Ambiente 2231 to the senior author and exported legally. Carlos Machado (Capibara Films, Mérida) recorded the species call with professional equipment. Photographs of the holotype of Pristimantis pleurostriatus are used thanks to the courtesy of Fernando Bird from the Universidad de Puerto Rico at Mayaguez. Kevin Zippel (Amphibian Ark) was kind enough to believe in the Fundación AndígenA’s work and helped support conservation work in Latin America. We thank Anna Vranos, who made a significant contribution to the Fundación AndígenA through the Amphibian Ark to fund this fieldwork. Finally we are deeply thankful to Anabel Rial from Conservation International Venezuela, who invited the senior author on the RAP to Calderas, where P. conservatio was discovered. In Calderas Oscar Lasso-Alcal and Jaime Bautista were great field companions. Funding for molecular work was provided by NSF to S.B.H.

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APPENDIX 1. Specimens examined.

Pristimantis boconoensis : CVULA 8371–72, Páramo de Boconó, Estado Trujillo, Venezuela. Pristimantis briceni : CVULA 2752–56, Páramo La Culata, CVULA 8364–65, trail from Monterrey to Cabaña del Cura, 2600 m, Sierra de la Culata, Cordillera de Mérida, Estado Mérida, Venezuela. Pristimantis conservatio : see type series. Pristimantis ginesi : CVULA 1567, Páramo de Mucubaji; 5950–59, Laguna Santo Cristo, Sierra Nevada, Estado Mérida, Venezuela. Pristimantis lancinii . CVULA 5578–79, La Corcovada; 2622, 5822, Páramo de Mucubají, Estado Mérida, Venezuela. Pristimantis paramerus : CVULA 766– 67, 990–93, 1003–05, 1859–60, 1994, 5828–34, 5836, 5838–48, 5850–54, La Corcovada, Paramo de Mucubaji, Estado Mérida, Venezuela. Pristimantis pleurostriatus . See referred specimens. The holotype UPR 4971 from San Eusebio, La Carbonera, Estado Mérida, Venezuela was photographed for us by F. Bird). Pristimantis rhigophilus : CVULA 6854, Páramo de Guaramacal, Estado Trujillo, Venezuela. Pristimantis vanadisae : CVULA 6169–70, Pozo Azul, Los Curos, Estado Mérida; CVULA 0 185, 0 283, 2191, 4926–27, 6084: La Carbonera, Estado Mérida. CVULA 0 632, 2353: San Javier del Valle, La Culata, Estado Mérida. CVULA 3107: Via El Morro, Estado Mérida. CVULA 0 186, 0 285, 1116–24, 1163–67, 1234–35, 1642– 55, 1681–90, 2014, 2317, 2016–23, 2156–57, 2208–09, 2354–62, 2605: Monte Zerpa, NW of the city of Mérida Estado Mérida. CVULA 0745–47, 3108–21, 3123–27: La Mucuy Alta, Tabay. CVULA 7184–85: Altos de San Luis, La Azulita, Estado Mérida. CVULA7194–96, 7198–206, 7206 from La Bravera, way from Mérida to La Azulita, 2200 m, Estado Mérida.

APPENDIX 2. Additional specimens newly sequenced for phylogenetic analyses.

Pristimantis aff. briceni . CVULA 8352: Piñango village, 2400 m. CVULA 8353: Páramo de Piñango, 3400 m. Pristimantis briceni . CVULA 8364: trail from Monterrey to Cabaña del Cura, 2600 m, Sierra de la Culata, Cordillera de Mérida, Estado Mérida.

Pristimantis ginesi . CVULA 8363: way from Laguna Mucubaj to Laguna Negra, Estado Mérida. Pristimantis lancinii . CVULA 8354, 8357: La Motús Alta, 3200 m. CVULA 8358, SBH 268517: Mucubaj. CVULA 8362: Quebrada del Padre, Estado Mérida. Pristimantis paramerus . (to be deposited; field ID number CB 11) La Corcovada, Paramo de Mucubají, Estado Mérida. Pristimantis aff. vanadisae . JX155281 CVULA7184, Finca Cedral, Altos de San Luis, Estado Mérida. Pristimantis vanadisae . SBH 268524: cloud forest above Truchicultura La Mucuy, Estado Mérida.

Figures

FIGURE 8. Maximum likelihood phylogeny of strabomantid frogs, rooted with Craugastor podiciferus. Bootstrap support values (likelihood / parsimony) are indicated at nodes. Pattern variants of Pristimantis vanadisae are noted. Museum, or when not available, field voucher numbers (for newly sequenced specimens) and GenBank accession numbers (bold; for all taxa in tree) are given. For newly-sequenced specimens, locality data are provided in species accounts, Appendix 1, or Appendix 2.