The new species was observed only at night during the rain. At the type locality, specimens were found in a grass-field near temporary pools.
A Monograph of the Frogs of the Family Microhylidae - PDF Free Download
The sequence divergences between Microhyla nilphamariensis sp. Intraspecific genetic divergence within the new species was estimated at 0. Molecular phylogeny suggests that M. Analysis is based on bp of mtDNA 16S gene sequence, showing the position of Microhyla nilphamariensis sp. Chaperina fusca was used as an outgroup. Numbers on branches represent bootstrap support values for Maximum-likelihood, and Bayesian posterior probabilities, respectively.
Quantitative diagnostic characters include see also: Table 1 , S2 Fig. Nevertheless, that the M. Ventral views of foot A Microhyla nilphamariensis sp. Ventral views of palm of E Microhyla nilphamariensis sp. Genetic divergence of M. Likewise, the new species is morphologically different from all known Southeast Asian species in comparison to the original descriptions [ 39 — 55 ].
Morphological characters of the other Southeast Asian species M. Shovel-shaped inner metatarsal tubercle and a more rounded snout separate M. Absence of digital discs differentiates M. Microhyla ornata is considered as one of the most common Microhyla species in Bangladesh, exhibiting a high degree of morphological similarity with other species in the genus.
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Both Matsui et al. However, no formal descriptions or detailed morphological comparison with other congeneric species were provided for these candidate species. Morphology-based species descriptions are known to be problematic in the genus Microhyla because of the high likelihood of homoplasy [ 62 , 63 ].
Also, the minute body size of Microhyla species poses challenges in diagnosing them from their known congeners based on morphology [ 10 , 64 ]. However, species in this genus are often strongly differentiated in genetic comparisons, facilitating identification of new candidate species. In our study, we have identified a new species that can be clearly differentiated from all know species in the genus Microhyla , both by detailed morphological comparisons and by genetic methods.
Hence, on the basis of a high degree of genetic divergence and subtle but clear-cut phenotypic divergence from M. The newly described species grouped genetically with unidentified haplotypes reported as new candidate species by Matsui et al. In fact, GenBank contains several sequences designated as M. After aligning all the sequences assigned to M. Interestingly, we also found sequences designated to M.
This indicates that the new species M. Therefore, further studies on the presence of the new species and its distribution in India are warranted. The new species lacks distinctive metatarsal tubercles, which were clearly present in the M. In addition to metatarsal tubercles, the large and pointed snout of M. Hence, the new species is differentiated from M. However, since the original author did not designate the M. This is because they bear the closest morphological similarity with the original description, and also because they originate from the type locality and regions close to it in the Western Ghats of India.
In addition, specimens of M.
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The use of a highly divergent outgroup for phylogenetic analysis may lead to errors because as the distance from the root to the ingroup increases, the shared character states between the divergent group and ingroup taxa may not be based on history, but to chance [ 66 ]. For lack of a better alternative, we used a rather divergent outgroup from the genus Gastrophryne Family Microhylidae to calibrate one of the nodes needed for estimation of the divergence times between taxa in our phylogenetic analysis.
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Although the Gastrophryne fossil provides a reliable estimate for calibration of the divergence time, it may be problematic in being highly divergent from the ingroup taxa. Nevertheless, we also calibrated the divergence time between the ingroup nodes using the known divergence between Microhyla fissipes and M. The analyses show that the new species diverged from M.
As India and the Bengal basin were the first to contact Southeast Asia in the early Miocene 22 mya [ 67 ], this corresponds well with our molecular clock analyses that indicated that the South Asian Microhylids diverged from the other congeneric species about 23 mya ago. Calibration points are indicated with arrows. In conclusion, we have described a new species of Microhyla from a highly genetically heterogeneous group of frogs that have been recognized as M. To this end, our study serves as an example of how detailed genetic and morphological comparisons among populations of species reportedly having a very broad distribution range can help in identification of yet undescribed species.
This in turn can provide valuable information for assigning correct conservation status for these taxa.
As to the status of M. Hence, M. Therefore, detailed morphological and genetic analyses would be in place to further resolve taxonomic uncertainties and identifying the cryptic diversity within this genus. See Materials and methods for explanation of trait abbreviations.
A distance from front of eyes to the nostril EN vs. GenBank accession numbers and locality information is included after the scientific names. The star marked haplotype for Microhyla ornata is from the type locality Kerala, India included in the present study. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.
National Center for Biotechnology Information , U. PLoS One. Published online Mar Sujith V. Author information Article notes Copyright and License information Disclaimer. Competing Interests: The authors have declared that no competing interests exist. Received Jun 5; Accepted Feb 3. This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are properly credited. This article has been cited by other articles in PMC.
Associated Data Supplementary Materials S1 Fig: A schematic illustration of the definitions of morphological traits measured in this study. S2 Fig: Bivariate plots of some diagnostic traits Microhyla nilphamariensis sp. S3 Fig: Maximum-likelihood phylogenetic tree based on variation in 16S gene fragment showing the position of Microhyla nilphamariensis sp. S1 Table: Additional specimens examined. S2 Table: Gene sequences compared and deposited in GenBank. Abstract A new species of Microhyla frog from the Nilphamari district of Bangladesh is described and compared with its morphologically similar and geographically proximate congeners.
Introduction Species delimitation is an important element in both ecology and evolutionary biology research, and in particular, in the development of biodiversity management strategies and plans [ 1 ]. Taxa and specimens Seven adult specimens one male and seven females of Microhyla sp. Open in a separate window.
Fig 1. A map showing localities from where Microhyla nilphamariensis sp. Morphological measurements and analyses Measurements were taken with digital calipers to the nearest 0. Nomenclatural Acts The electronic edition of this article conforms to the requirements of the amended International Code of Zoological Nomenclature ICZN , and hence the new names contained herein are available under that Code from the electronic edition of this article.
Diagnosis Microhyla nilphamariensis sp. Fig 2.
A monograph of the frogs of the family Microhylidae
Photographs of Microhyla nilphamariensis sp. Measurements in mm Male holotype : SVL Variation Morphometric variability is described in Table 1. Table 1 Summary of quantitative and qualitative diagnostic characters in Microhyla nilphamariensis sp. Elongated inner metacarpal tubercle; very large and prominent, heart-shaped outer metatarsal tubercle. Metatarsal tubercle Inner metatarsal tubercle small round shaped; outer metatarsal tubercle is ovoid, minute, and indistinct. Inner metatarsal tubercle elongated, large and very prominent; outer metatarsal tubercle is compressed and large in size.
Inner metatarsal tubercle shovel-shaped, bearing longitudinal groove, large and prominent; outer metatarsal tubercle is also shovel-shaped and large. Distribution Microhyla nilphamariensis sp. Natural history The new species was observed only at night during the rain. Molecular phylogeny and genetic divergence of new species The sequence divergences between Microhyla nilphamariensis sp. Table 2 Pairwise genetic divergence number of base substitutions per site among Microhyla species based on bp mtDNA 16S gene sequences. Holotype b M. Fig 3.
Phylogenetic relationships among all known species in the genus Microhyla.
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