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      Noblella thiuni sp. n., a new (singleton) species of minute terrestrial-breeding frog (Amphibia, Anura, Strabomantidae) from the montane forest of the Amazonian Andes of Puno, Peru

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      PeerJ

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      Frog, Taxonomy, Carabaya, 16S rRNA, Cloud forest, Ollachea, Leaf litter amphibian

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          Abstract

          We describe a new species of minute, terrestrial-breeding frog in the genus Noblella. We collected a single specimen in the leaf litter of primary montane forest (2,225 m a.s.l.) near Thiuni, in the Provice of Carabaya, Department of Puno, in the upper watershed of a tributary of the Inambari River of southern Peru, the same locality where we found the types of Psychrophrynella glauca Catenazzi & Ttito 2018. We placed the new species within Noblella on the basis of molecular data, minute size, and overall morphological resemblance with the type species N. peruviana and other species of Noblella, including having three phalanges on finger IV (as in N. coloma, N. heyeri, N. lynchi, N. madreselva, N. peruviana, and N. pygmaea), and terminal phalanges T-shaped and pointed. Noblella thiuni sp. n. is distinguished from all other species of Noblella by having ventral surfaces of legs bright red, and chest and belly copper reddish with a profusion of silvery spots. The new species further differs from known Peruvian species of Noblella by the combination of the following characters: tympanic membrane absent, eyelids lacking tubercles, dorsal skin finely shagreen, tarsal tubercles or folds absent, three phalanges on Finger IV, tips of digits not expanded, no circumferential grooves on digits, inguinal spots present. The new species has a snout–vent length of 11.0 mm in one adult or subadult male. Our new finding confirms the high levels of endemism and beta diversity of small, terrestrial-breeding frogs inhabiting the moss layers and leaf litter in the montane forests of the Amazonian slopes of the Andes and adjacent moist puna grasslands, and suggests much work remains to be done to properly document this diversity.

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          Most cited references 31

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          The integrative future of taxonomy

          Background Taxonomy is the biological discipline that identifies, describes, classifies and names extant and extinct species and other taxa. Nowadays, species taxonomy is confronted with the challenge to fully incorporate new theory, methods and data from disciplines that study the origin, limits and evolution of species. Results Integrative taxonomy has been proposed as a framework to bring together these conceptual and methodological developments. Here we review perspectives for an integrative taxonomy that directly bear on what species are, how they can be discovered, and how much diversity is on Earth. Conclusions We conclude that taxonomy needs to be pluralistic to improve species discovery and description, and to develop novel protocols to produce the much-needed inventory of life in a reasonable time. To cope with the large number of candidate species revealed by molecular studies of eukaryotes, we propose a classification scheme for those units that will facilitate the subsequent assembly of data sets for the formal description of new species under the Linnaean system, and will ultimately integrate the activities of taxonomists and molecular biologists.
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            Underestimation of Species Richness in Neotropical Frogs Revealed by mtDNA Analyses

            Background Amphibians are rapidly vanishing. At the same time, it is most likely that the number of amphibian species is highly underestimated. Recent DNA barcoding work has attempted to define a threshold between intra- and inter-specific genetic distances to help identify candidate species. In groups with high extinction rates and poorly known species boundaries, like amphibians, such tools may provide a way to rapidly evaluate species richness. Methodology Here we analyse published and new 16S rDNA sequences from 60 frog species of Amazonia-Guianas to obtain a minimum estimate of the number of undescribed species in this region. We combined isolation by distance, phylogenetic analyses, and comparison of molecular distances to evaluate threshold values for the identification of candidate species among these frogs. Principal Findings In most cases, geographically distant populations belong to genetically highly distinct lineages that could be considered as candidate new species. This was not universal among the taxa studied and thus widespread species of Neotropical frogs really do exist, contrary to previous assumptions. Moreover, the many instances of paraphyly and the wide overlap between distributions of inter- and intra-specific distances reinforce the hypothesis that many cryptic species remain to be described. In our data set, pairwise genetic distances below 0.02 are strongly correlated with geographical distances. This correlation remains statistically significant until genetic distance is 0.05, with no such relation thereafter. This suggests that for higher distances allopatric and sympatric cryptic species prevail. Based on our analyses, we propose a more inclusive pairwise genetic distance of 0.03 between taxa to target lineages that could correspond to candidate species. Conclusions Using this approach, we identify 129 candidate species, two-fold greater than the 60 species included in the current study. This leads to estimates of around 170 to 460 frog taxa unrecognized in Amazonia-Guianas. Significance As a consequence the global amphibian decline detected especially in the Neotropics may be worse than realised.
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              Deciphering amphibian diversity through DNA barcoding: chances and challenges.

              Amphibians globally are in decline, yet there is still a tremendous amount of unrecognized diversity, calling for an acceleration of taxonomic exploration. This process will be greatly facilitated by a DNA barcoding system; however, the mitochondrial population structure of many amphibian species presents numerous challenges to such a standardized, single locus, approach. Here we analyse intra- and interspecific patterns of mitochondrial variation in two distantly related groups of amphibians, mantellid frogs and salamanders, to determine the promise of DNA barcoding with cytochrome oxidase subunit I (cox1) sequences in this taxon. High intraspecific cox1 divergences of 7-14% were observed (18% in one case) within the whole set of amphibian sequences analysed. These high values are not caused by particularly high substitution rates of this gene but by generally deep mitochondrial divergences within and among amphibian species. Despite these high divergences, cox1 sequences were able to correctly identify species including disparate geographic variants. The main problems with cox1 barcoding of amphibians are (i) the high variability of priming sites that hinder the application of universal primers to all species and (ii) the observed distinct overlap of intraspecific and interspecific divergence values, which implies difficulties in the definition of threshold values to identify candidate species. Common discordances between geographical signatures of mitochondrial and nuclear markers in amphibians indicate that a single-locus approach can be problematic when high accuracy of DNA barcoding is required. We suggest that a number of mitochondrial and nuclear genes may be used as DNA barcoding markers to complement cox1.
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                Author and article information

                Contributors
                Journal
                PeerJ
                PeerJ
                peerj
                peerj
                PeerJ
                PeerJ Inc. (San Diego, USA )
                2167-8359
                23 April 2019
                2019
                : 7
                Affiliations
                [1 ]Biological Sciences, Florida International University , Miami, FL, United States of America
                [2 ]Centro de Ornitología y Biodiversidad , Lima, Perú
                [3 ]Departamento de Ecología, Facultad de Ciencias Biológicas, Pontificia Universidad Católica de Chile , Santiago, Chile
                [4 ]Museo de Historia Natural, Universidad Nacional de San Antonio Abad , Cusco, Perú
                Article
                6780
                10.7717/peerj.6780
                6485238
                8c067381-2827-42aa-93e8-3a7d8f8d99cc
                ©2019 Catenazzi and Ttito

                This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, reproduction and adaptation in any medium and for any purpose provided that it is properly attributed. For attribution, the original author(s), title, publication source (PeerJ) and either DOI or URL of the article must be cited.

                Funding
                Funded by: The Eppley Foundation
                Funded by: Wildlife Acoustics
                Funded by: Chicago Board of Trade Endangered Species Fund
                This work was supported by grants from the Eppley Foundation, Wildlife Acoustics, and the Chicago Board of Trade Endangered Species Fund. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.
                Categories
                Biodiversity
                Conservation Biology
                Evolutionary Studies
                Taxonomy
                Zoology

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