A systematic revision of the genus Hylarana in the Western Ghats-Sri Lanka biodiversity hotspot is presented. Species delineation in Hylarana is complicated due to a lack of distinct colour differences or striking morphological characters, leading to potential misidentification. We conducted extensive surveys throughout the Western Ghats-Sri Lanka biodiversity hotspot and performed multiple gene (16S, COI and Cytb) barcoding using 103 samples collected from cultivated land and natural habitats. Genetic distance comparisons and Neighbor Joining trees indicated the presence of at least 14 candidate species in the region, supported by taxa groupings for all three genetic markers. Utilising a combination of molecular and morphological data, we describe seven new species, doubling the number of Hylarana species previously known from this region. We further demonstrate that H. temporalis, which was originally described from Sri Lanka, was misidentified with the Western Ghats endemic species for nearly 100 years. Conversely, H. aurantiaca was originally described from the Western Ghats and misidentified in Sri Lanka. Our study confirms that the distribution of H. temporalis is restricted to Sri Lanka, while H. aurantiaca is endemic to the Western Ghats, and that there are no shared Hylarana species between the two regions. Hylarana flavescens, H. intermedius and H. montanus, previously considered synonyms of H. temporalis are confirmed as valid species. Hylarana bhagmandlensis is removed from the synonymy of H. aurantiaca and placed as a junior subjective synonym of H. montanus. To establish nomenclatural stability, H. flavescens, H. malabarica and H. temporalis are lectotypified and H. intermedius is neotypified. Detailed descriptions, diagnosis, morphological and genetic comparisons, illustrations and data on distribution and natural history are provided for all species. Phylogenetic analyses based on three mitochondrial markers (16S, COI and Cytb) and a fragment of the nuclear Rag1 gene, show complete endemism of the Western Ghats-Sri Lankan species. Four major groups in this region are identified as: 1 — the Hylarana aurantiaca group, endemic to the Western Ghats; 2 — the Hylarana flavescens group, endemic to the Western Ghats; 3 — the Hylarana temporalis group, endemic to Sri Lanka; and 4 — the Hylarana malabarica group from Sri Lanka and India. The discovery of numerous morphologically cryptic Hylarana species in this region further emphasizes the benefits of utilizing an integrative taxonomic approach for uncovering hidden diversity and highlighting local endemism in the Western Ghats-Sri Lanka biodiversity hotspot.
Based on a recent molecular phylogenetic study, the South Asian microhylid genus Uperodon (subfamily Microhylinae) currently comprises of 12 valid species that are largely restricted to India and Sri Lanka. Considering the revised generic-level status of its various members, here we review the taxonomy of all known species in this genus and clarify their nomenclatural status and geographical distribution, by integrating evidence from genetics, adult and tadpole morphology, breeding ecology, and bioacoustics. Our molecular analyses of a mitochondrial 16S rRNA gene fragment combined with external and internal morphological studies also revealed a distinct new species in the genus. This species, formally described as Uperodon rohani sp. nov., is endemic to Sri Lanka and widely distributed at lower elevations in the island. For nomenclatural stability of various previously known members, the following actions are also undertaken: (1) redescription of the poorly-defined species Ramanella anamalaiensis Rao (= Uperodon anamalaiensis) and Hylaedactylus montanus Jerdon (= Uperodon montanus); (2) neotype designation for Ramanella anamalaiensis Rao (= Uperodon anamalaiensis), Ramanella minor Rao, Ramanella mormorata Rao (= Uperodon mormorata), and Ramanella triangularis rufeventris Rao; (3) lectotype designation for Callula variegata Stoliczka (= Uperodon variegatus); and (4) synonymization of Ramanella minor Rao with Uperodon anamalaiensis.
Species boundaries of Microhyla rubra of India and Sri Lanka were assessed using the following criteria: genetic barcoding, morphology, and vocalization. We use a ca. 500 bp fragment of the 16S rRNA mitochondrial gene and show that there is an uncorrected pairwise distance of 2.7-3.2% between the Indian and Sri Lankan populations of M. rubra. We show that they are different in several call characteristics such as, dominant frequency, call duration, call rise time and pulse rate. Morphologically, the Sri Lankan population can be distinguished from the typical M. rubra described from southern India, by a combination of characters: body size, skin texture, and feet dimensions. We recognize the population from Sri Lanka as a new species, Microhyla mihintalei sp. nov., a widely distributed lowland species with an elevational distribution of up to 500 m a.s.l.
The bufonid genus Adenomus, an endemic of the montane and lowland rainforests of central and south-western Sri Lanka, has been considered to comprise of three species, viz. A. kelaartii, A. dasi and A. kandianus, the last of which has been recently highlighted as "the world's rarest toad". We conducted a survey across the known range of Adenomus and used multiple criteria to delineate species boundaries within the genus. These include: a molecular phylogeny based on a 16S ribosomal RNA gene fragment; an examination of the external morphology of adults and larvae, and the skeletal morphology of adults; a bioacoustic analysis; and ecological niche modelling. We show that Adenomus is monophyletic and that it comprises only two species: A. kelaartii and A. kandianus, with A. dasi being a junior synonym of the latter. For the two valid species of Adenomus, we provide detailed osteological descriptions; clarify the distribution patterns; and provide genetic data to facilitate their scientific conservation management.
The monotypic genus Lankanectes, considered an evolutionary long branch with India’s Nyctibatrachus as its sister lineage, is represented by L. corrugatus, a species widely distributed within the wet zone of Sri Lanka up to 1500 m asl, where it inhabits a variety of lotic and lentic habitats. Here, following an integrative taxonomic approach using DNA-based phylogenies, morphology, morphometry, and ecological niche models, we describe a new species—Lankanectes pera sp. nov. The new species is distinguished from its sister species mainly by its tuberculated throat and absence of dark patches on venter, throat, manus and pes. The uncorrected genetic distances between the two Lankanectes species for a fragment of the non-coding mitochondrial 16S rRNA gene is 3.5–3.7%. The new species has a very restricted climatic distribution with a total predicted area of only 360 km2 (vs. 14,120 km2 for L. corrugatus). Unlike L. corrugatus, which prefers muddy substrates and marshy areas, the new species is observed inhabiting only pristine streams flowing through canopy covered montane forests in the highest reaches of the Knuckles Mountain range. The specialized new species will need immediate conservation attention due to its restricted distribution (montane isolate), specialized habit of inhabiting clear mountain streams, and small population size.
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