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Genetic diversity is frequently described using heterozygosity, particularly in a conservation context. Often, it is estimated using single nucleotide polymorphisms (SNPs); however, it has been shown that heterozygosity values calculated from SNPs can be biased by both study design and filtering parameters. Though solutions have been proposed to address these issues, our own work has found them to be inadequate in some circumstances. Here, we aimed to improve the reliability and comparability of heterozygosity estimates, specifically by investigating how sample size and missing data thresholds influenced the calculation of autosomal heterozygosity (heterozygosity calculated from across the genome, i.e. fixed and variable sites). We also explored how the standard practice of tri- and tetra-allelic site exclusion could bias heterozygosity estimates and influence eventual conclusions relating to genetic diversity. Across three distinct taxa (a frog, Litoria rubella; a tree, Eucalyptus microcarpa; and a grasshopper, Keyacris scurra), we found heterozygosity estimates to be meaningfully affected by sample size and missing data thresholds, partly due to the exclusion of tri- and tetra-allelic sites. These biases were inconsistent both between species and populations, with more diverse populations tending to have their estimates more severely affected, thus having potential to dramatically alter interpretations of genetic diversity. We propose a modified framework for calculating heterozygosity that reduces bias and improves the utility of heterozygosity as a measure of genetic diversity, whilst also highlighting the need for existing population genetic pipelines to be adjusted such that tri- and tetra-allelic sites be included in calculations., (© 2024 The Authors. Molecular Ecology Resources published by John Wiley & Sons Ltd.)

Litoria rothii is a widespread pelodryadid frog with a charismatic laughing advertisement call, distributed across the Australian Monsoon Tropics and southern New Guinea. Given its large distribution spanning well-known biogeographic barriers, variation in male advertisement calls and the prevalence of unresolved species complexes in the Australian frog fauna, we examine the genetic, morphological and acoustic diversity in the species from across its range. Our analyses reveal the presence of a previously unrecognised species in western parts of the range of L. rothii sensu lato, which we describe herein as a new species. Litoria ridibunda sp. nov. is distinguished from L. rothii on the basis of paraphyly of nuclear gene trees with L. everetti from Indonesia, colour patterns on the posterior thigh and male advertisement calls. Compared to L. rothii, the new species has a less contrasting pattern on the posterior thigh and a male advertisement call with a greater number of notes per call and a greater call duration. In particular, the magnitude of call differences between the species is highest where the ranges of the two species are in proximity in north-western Queensland. Our study further emphasises the undiagnosed diversity that remains in Australian frogs, even in relatively large, charismatic, frequently encountered species that often share human dwellings.

When closely related species come into contact via range expansion, both may experience reduced fitness as a result of the interaction. Selection is expected to favour traits that minimize costly interspecies reproductive interactions (such as mismating) via a phenomenon called reproductive character displacement (RCD). Research on RCD frequently assumes secondary contact between species, but the geographical history of species interactions is often unknown. Population genomic data permit tests of geographical hypotheses about species origins and secondary contact through range expansion. We used population genomic data from single nucleotide polymorphisms (SNPs), mitochondrial sequence data, advertisement call data and morphological data to investigate a species complex of toadlets (Uperoleia borealis, U. crassa, U. inundata) from northern Australia. Although the three species of frogs were morphologically indistinguishable in our analysis, we determined that U. crassa and U. inundata form a single species (synonymized here) based on an absence of genomic divergence. SNP data identified the phylogeographical origin of U. crassa as the Top End, with subsequent westward invasion into the range of U. borealis in the Kimberley. We identified six F 1 hybrids, all of which had the U. borealis mitochondrial haplotype, suggesting unidirectional hybridization. Consistent with the RCD hypothesis, U. borealis and U. crassa sexual signals differ more in sympatry than in allopatry. Hybrid males have intermediate calls, which probably reduces attractiveness to females. Integrating population genomic data, mitochondrial sequencing, morphology and behavioural approaches provides an unusually detailed collection of evidence for reproductive character displacement following range expansion and secondary contact., (© 2022 The Authors. Molecular Ecology published by John Wiley & Sons Ltd.)

Many Drosophila species differ widely in their distributions and climate niches, making them excellent subjects for evolutionary genomic studies. Here, we have developed a database of high-quality assemblies for 46 Drosophila species and one closely related Zaprionus. Fifteen of the genomes were newly sequenced, and 20 were improved with additional sequencing. New or improved annotations were generated for all 47 species, assisted by new transcriptomes for 19. Phylogenomic analyses of these data resolved several previously ambiguous relationships, especially in the melanogaster species group. However, it also revealed significant phylogenetic incongruence among genes, mainly in the form of incomplete lineage sorting in the subgenus Sophophora but also including asymmetric introgression in the subgenus Drosophila. Using the phylogeny as a framework and taking into account these incongruences, we then screened the data for genome-wide signals of adaptation to different climatic niches. First, phylostratigraphy revealed relatively high rates of recent novel gene gain in three temperate pseudoobscura and five desert-adapted cactophilic mulleri subgroup species. Second, we found differing ratios of nonsynonymous to synonymous substitutions in several hundred orthologues between climate generalists and specialists, with trends for significantly higher ratios for those in tropical and lower ratios for those in temperate-continental specialists respectively than those in the climate generalists. Finally, resequencing natural populations of 13 species revealed tropics-restricted species generally had smaller population sizes, lower genome diversity and more deleterious mutations than the more widespread species. We conclude that adaptation to different climates in the genus Drosophila has been associated with large-scale and multifaceted genomic changes., (© 2021 The Authors. Molecular Ecology Resources published by John Wiley & Sons Ltd.)

There is a high rate of recent species discovery in remote regions of northern Australia, especially for amphibians and reptiles. The Wessel Islands, located in the northeastern corner of the Northern Territory, has recently been identified as a region of high species and phylogenetic endemism based on samples collected during the sole reptile and amphibian survey there in 1993. Using a phylogenetic approach, we describe a new, evolutionarily distinct species of Uperoleia endemic to the Wessel Islands. This description is based on three specimens, one female and two juveniles, which represent the only confirmed vouchers of the species. Due to the low number of specimens, this new species cannot be diagnosed morphologically from other closely related Uperoleia, and nothing is currently known about the mating call or basic biology. The discovery of this species provides further evidence for the islands importance as an area of endemism and identifies an urgent need for further surveys to document the unique biological diversity of the Wessel Islands.

The Queensland fruit fly ( Bactrocera tryoni ; Q-fly) is an Australian endemic horticultural pest species, which has caused enormous economic losses. It has the potential to expand its range to currently Q-fly-free areas and poses a serious threat to the Australian horticultural industry. A large number of studies have investigated the correlation between environmental factors and Q-fly development, reproduction, and expansion. However, it is still not clear how Q-fly morphological traits vary with the environment. Our study focused on three morphological traits (body size, wing shape, and fluctuating asymmetry) in Q-fly samples collected from 1955 to 1965. We assessed how these traits vary by sex, and in response to latitude, environmental variables, and geographic distance. First, we found sexual dimorphism in body size and wing shape, but not in fluctuating asymmetry. Females had a larger body size but shorter and wider wings than males, which may be due to reproductive and/or locomotion differences between females and males. Secondly, the body size of Q-flies varied with latitude, which conforms to Bergmann's rule. Finally, we found Q-fly wing shape was more closely related to temperature rather than aridity, and low temperature and high aridity may lead to high asymmetry in Q-fly populations.

Molecular phylogenetics plays a key role in comparative genomics and has increasingly significant impacts on science, industry, government, public health and society. In this paper, we posit that the current phylogenetic protocol is missing two critical steps, and that their absence allows model misspecification and confirmation bias to unduly influence phylogenetic estimates. Based on the potential offered by well-established but under-used procedures, such as assessment of phylogenetic assumptions and tests of goodness of fit, we introduce a new phylogenetic protocol that will reduce confirmation bias and increase the accuracy of phylogenetic estimates., (© The Author(s) 2019. Published by Oxford University Press on behalf of NAR Genomics and Bioinformatics.)

Understanding how natural populations will respond to rapid anthropogenic climate change is one of the greatest challenges for ecologists and evolutionary biologists. Much research has focussed on whether physiological traits can evolve quickly enough under rapidly increasing temperatures. While the simple Breeder's equation helps to understand how extreme temperatures and genetic variation might drive within-population evolution under climate change, it does not consider two key areas: how different forms of phenotypic plasticity interact and variation among populations. Plasticity can modify the exposure to climatic extremes and the strength of selection from those extremes, while differences among populations provide adaptive diversity not apparent within them. Here, we focus on terrestrial vertebrates and, with a case study on a tropical lizard, demonstrate the complex interplay between spatial, genetic and plastic contributions to variation in climate-relevant physiological traits. We identify several problems that need to be better understood: which traits are under selection in a changing climate; the different forms of plasticity relevant to population persistence and rapid evolution; plastic versus genetic contributions to geographic variation in climate-associated traits and whether plasticity can be harnessed to promote persistence of species. Given ongoing uncertainties around whether natural populations can evolve rapidly enough to persist, we advocate the use of field trials aimed at increasing rates of adaptation, especially in systems known to be strongly impacted by human-driven changes in climate., (Copyright © 2019 Elsevier Ltd. All rights reserved.)

Testing hypotheses on drivers of clade evolution and trait diversification provides insight into many aspects of evolutionary biology. Often, studies investigate only intrinsic biological properties of organisms as the causes of diversity, however, extrinsic properties of a clade's environment, particularly geological history, may also offer compelling explanations. The Andes are a young mountain chain known to have shaped many aspects of climate and diversity of South America. The Liolaemidae are a radiation of South American reptiles with over 300 species found across most biomes and with similar numbers of egg-laying and live-bearing species. Using the most complete dated phylogeny of the family, we tested the role of Andean uplift in biogeography, diversification patterns, and parity mode of the Liolaemidae. We find that the Andes promoted lineage diversification and acted as a species pump into surrounding biomes. We also find strong support for the role of Andean uplift in boosting the species diversity of these lizards via allopatric fragmentation. Finally, we find repeated shifts in parity mode associated with changing thermal niches, with live-bearing favored in cold climates and egg-laying favored in warm climates. Importantly, we find evidence for possible reversals to oviparity, an evolutionary transition believed to be extremely rare., (© 2018 The Author(s). Evolution © 2018 The Society for the Study of Evolution.)

The spectacular threat display of the savannah specialist Australo-Papuan frilled lizards has made them one of the world's most iconic reptiles. They are increasingly used as a model system for research in evolutionary biology and ecology but little is known of their population structure. Their distribution across northern Australia and southern New Guinea also provides an opportunity to examine biogeographic patterns as they relate to the large-scale movement of savannah habitat during the Plio/Pleistocene and the associated increase in aridity. We generated sequence data for one mitochondrial and four nuclear DNA loci (5052 base pairs) for 83 frilled lizards sampled throughout their range. We also quantified body proportion variation for 279 individuals. Phylogenetic analyses based on maximum likelihood and Bayesian species-tree methods revealed three shallow clades that replace each other across the monsoon tropics. We found the expected pattern of male biased sexual size dimorphism in both maximum body size and head size but there was no sexual dimorphism in overall body shape or in frill size, relative to head size, supporting the hypothesis that the frill is used primarily as a threat display rather than a sexual display. The genetic clades are broadly consistent with known clinal variation in frill color that gradually shifts from west to east (red, orange, yellow/white) but otherwise show little morphological differentiation in body proportion measures. The biogeographic breaks between clades occur at the Carpentaria Gap and the lowlands surrounding the Ord River, and our ecological niche modeling predicts lower habitat suitability for C. kingii in these regions. While this biogeographic pattern is consistent with numerous other taxonomic groups in northern Australia, the overall low genetic diversity in frilled lizards across the entire monsoon tropics and southern New Guinea contrasts starkly to patterns seen in other terrestrial vertebrates. Extremely low intra-clade genetic diversity over vast geographic areas is indicative of recent gene flow that would likely have been facilitated by widespread savannah during interglacials, or alternatively may reflect population bottlenecks induced by extreme aridity during Pleistocene glacials. The shallow divergence between Australian and New Guinean samples is consistent with recent connectivity between Australia and New Guinea that would have been possible via a savannah corridor across the Torres Strait. Based on our molecular and morphological data, we do not support taxonomic recognition of any of the frilled lizard clades and instead consider C. kingii a single species with shallow phylogeographic structure and clinal variation in frill color., (Copyright © 2016 Elsevier Inc. All rights reserved.)

The discovery of new vertebrate species in developed countries is still occurring at surprising rates for some taxonomic groups, especially the amphibians and reptiles. While this most often occurs in under-explored areas, it occasionally still happens in well-inhabited regions. We report such a case with the discovery and description of U. mahonyi sp. nov., a new species of frog from a highly populated region of New South Wales, Australia. We provide details of its morphology, calls, embryos and tadpoles, and phylogenetic relationships to other species of eastern Uperoleia. We also provide the results of targeted surveys to establish its distribution and provide observations of its habitat associations. As a consequence of these surveys, we comment on the likely restricted nature of the species' distribution and habitat, and place this in the context of a preliminary assessment of its putative conservation status, which should be assessed for listing under the IUCN's red list. We note this species, which is morphologically distinct, has gone unnoticed for many decades despite numerous ecological surveys for local development applications.

Areas of suitable habitat for species and communities have arisen, shifted, and disappeared with Pleistocene climate cycles, and through this shifting landscape, current biodiversity has found paths to the present. Evolutionary refugia, areas of relative habitat stability in this shifting landscape, support persistence of lineages through time, and are thus crucial to the accumulation and maintenance of biodiversity. Areas of endemism are indicative of refugial areas where diversity has persisted, and endemism of intraspecific lineages in particular is strongly associated with late-Pleistocene habitat stability. However, it remains a challenge to consistently estimate the geographic ranges of intraspecific lineages and thus infer phylogeographic endemism, because spatial sampling for genetic analyses is typically sparse relative to species records. We present a novel technique to model the geographic distribution of intraspecific lineages, which is informed by the ecological niche of a species and known locations of its constituent lineages. Our approach allows for the effects of isolation by unsuitable habitat, and captures uncertainty in the extent of lineage ranges. Applying this method to the arc of rainforest areas spanning 3500 km in eastern Australia, we estimated lineage endemism for 53 species of rainforest dependent herpetofauna with available phylogeographic data. We related endemism to the stability of rainforest habitat over the past 120,000 years and identified distinct concentrations of lineage endemism that can be considered putative refugia. These areas of lineage endemism are strongly related to historical stability of rainforest habitat, after controlling for the effects of current environment. In fact, a dynamic stability model that allows movement to track suitable habitat over time was the most important factor in explaining current patterns of endemism. The techniques presented here provide an objective, practical method for estimating geographic ranges below the species level, and including them in spatial analyses of biodiversity.

Australia is a large and complex landmass that comprises diverse biomes ranging from tropical rainforests to harsh deserts. While Australian biotic diversity has evolved in response to landscape and climate changes, evidence of Miocene or later biome shifts are few. The Australo-Papuan endemic frog genus Uperoleia is widely distributed across mesic, monsoonal tropic and arid regions of Australia. Thus, it represents an ideal system to evaluate biome shifts as they relate to known landscape and climate history. We comprehensively sampled the distributional range of 25 described Uperoleia species and generated a detailed molecular phylogeny for the genus based on one mitochondrial and five nuclear loci. Our results support a single origin of monsoonal tropic taxa, followed by diversification within the region under the influence of the Australian monsoon. Molecular dating analyses suggest the major divergence between eastern mesic and monsoonal species occurred in the Miocene approximately 17million years ago, with repeated evolution of species from monsoonal biomes to arid or mesic biomes in the later Miocene, early Pliocene and at the beginning of the Pleistocene. Our detailed sampling helps to clarify the true distributions of species and contributes to on-going work to improve the taxonomy of the genus. Topological differences between nuclear and mitochondrial phylogenies within major clades suggest a history of mitochondrial introgression and capture, and reduce the ability to resolve close interspecific relationships., (Copyright © 2014 Elsevier Inc. All rights reserved.)

The frog genus Uperoleia (Myobatrachidae) is species rich, with the greatest diversity in the northern monsoonal region of Australia. Due in part to their small body size, conservative morphology and distribution in diverse habitats, the genus is likely to harbor cryptic species. A recent study (Catullo et al. 2013) assessed region-wide genetic, acoustic and phenotypic variation within four species in northern Australia. Catullo et al. (2013) presented multiple lines of evidence that the widespread U. trachyderma comprises distinct allopatric western and eastern lineages within the Northern Deserts bioregion of Australia. Here we formally describe the western lineage as U. stridera sp. nov. and redescribe the eastern (type) clade as U. trachyderma. The new species can be distinguished from U. trachyderma by fewer pulses per call, a faster pulse rate, and the lack of scattered orange to red flecks on the dorsum. The description of U. stridera sp. nov. brings the number of Uperoleia species to 28, by far the largest genus in the Myobatrachidae, and further highlights the Australian monsoonal tropics as a region of high endemism.