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Climatic and evolutionary processes are inextricably linked to conservation. Avoiding extinction in rapidly changing environments often depends upon a species' capacity to adapt in the face of extreme selective pressures. Here, we employed exon capture and high-throughput next-generation sequencing to investigate the mechanisms underlying population structure and adaptive genetic variation in the koala (Phascolarctos cinereus), an iconic Australian marsupial that represents a unique conservation challenge because it is not uniformly threatened across its range. An examination of 250 specimens representing 91 wild source locations revealed that five major genetic clusters currently exist on a continental scale. The initial divergence of these clusters appears to have been concordant with the Mid-Brunhes Transition (~430 to 300 kya), a major climatic reorganisation that increased the amplitude of Pleistocene glacial-interglacial cycles. While signatures of polygenic selection and environmental adaptation were detected, strong evidence for repeated, climate-associated range contractions and demographic bottleneck events suggests that geographically isolated refugia may have played a more significant role in the survival of the koala through the Pleistocene glaciation than in situ adaptation. Consequently, the conservation of genome-wide genetic variation must be aligned with the protection of core koala habitat to increase the resilience of vulnerable populations to accelerating anthropogenic threats. Finally, we propose that the five major genetic clusters identified in this study should be accounted for in future koala conservation efforts (e.g., guiding translocations), as existing management divisions in the states of Queensland and New South Wales do not reflect historic or contemporary population structure.

Paubrasilia echinata (Lam.) Gagnon, H. C. Lima & G. P. Lewis (“Pau Brasil”) is the national tree of Brazil and an endangered species endemic to the Brazilian Atlantic Forest. The extensive range of distribution, spanning over 2000 km distance, is matched by extensive plasticity in leaf morphology. Three morphotypes are commonly identified based on the size of the leaflets but it is unclear if they represent distinct taxa or a single polymorphic species. This study aims to clarify the taxonomic position of the three morphotypes to inform conservation decisions. A comprehensive morphometric study based on herbarium specimens from the entire distribution range of the species was coupled with genetic analyses of population structure using genotype-by-sequencing data. We found that the three morphotypes do not match separately evolving lineages. Rather, P. echinata is composed of five genetic lineages that are geographically structured, although we did find evidence of genetic admixture in two individuals. Leaflet size varied by over 35-fold and although morphological clustering generally matched the genetic patterns, there were some overlaps, highlighting the cryptic diversity within this group. Finally, our genetic and morphological results provide some evidence that cultivated trees from different states in Brazil seem to be most closely related to a genetic lineage from northern Brazil, which suggests that more care is needed to protect and preserve the overall genomic diversity of this highly endangered and iconic species.

Robins in the family Petroicidae are characteristic of the woodland bird community that is threatened in Australia as a result of habitat loss and fragmentation. Flame Robin (Petroica phoenicea) populations declined by 56% between 1980 and 2000, with habitat loss likely being the primary cause. Given that Flame Robins primarily breed at high elevation, populations may become more isolated due to anthropogenic change, resulting in increased inbreeding and loss of genetic diversity that may accelerate local extinction. We estimated the genetic structure and recent gene flow among four populations (n = 70 birds) of this vulnerable (NSWSC) species across a 670 km portion of its range in temperate south-eastern Australia using 14 genetic markers. We found no significant differences in genetic diversity amongst populations and little population structuring – only the northernmost population showing a weak signal of differentiation. However, we detected little recent migration between the northern and southern sites, possibly due to recent fragmentation. We conclude that habitat loss is a conservation concern for this Vulnerable species and further work and ongoing genetic monitoring is needed, particularly given high elevation breeding sites that are vulnerable in the face of a changing climate.

Funding: Biotechnology and Biological Sciences Research Council (BBSRC) (Grants BB/G023913/1 and BB/G023913/2 to C.R., and studentship to B.C.K.), the School of Biology at the University of St Andrews (studentships to M.P.S. and B.C.K.), and the Leverhulme Trust (Grant RPG-2015-273 to P.M.H.). Some animals fashion tools or constructions out of plant materials to aid foraging, reproduction, self-maintenance, or protection. Their choice of raw materials can affect the structure and properties of the resulting artifacts, with considerable fitness consequences. Documenting animals’ material preferences is challenging, however, as manufacture behavior is often difficult to observe directly, and materials may be processed so heavily that they lack identifying features. Here, we use DNA barcoding to identify, from just a few recovered tool specimens, the plant species New Caledonian crows (Corvus moneduloides) use for crafting elaborate hooked stick tools in one of our long-term study populations. The method succeeded where extensive fieldwork using an array of conventional approaches—including targeted observations, camera traps, radio-tracking, bird-mounted video cameras, and behavioral experiments with wild and temporarily captive subjects—had failed. We believe that DNA barcoding will prove useful for investigating many other tool and construction behaviors, helping to unlock significant research potential across a wide range of study systems. Publisher PDF

The unique geography and climate of the Qinghai-Tibet plateau (QTP) have impacted the formation of species in this area, with a large number of relatively young endemic species associated with the uplift of the QTP and Quaternary climatic changes. In our study, we used two chloroplast DNA fragments and seven nuclear genes to investigate the population genetic structure, species divergence and demographic history of two QTP endemic species, Juniperus erectopatens and J. microsperma as well as their close relatives, J. sabina and J. semiglobosa. Furthermore, we assessed the correlation between their divergence and the historical environmental changes of the QTP. *BEAST analysis revealed that a clade containing J. erectopatens and J. microsperma separated from the clade containing J. sabina and J. semiglobosa about 2.72 million years ago (Mya), whereas the divergence between J. erectopatens and J. microsperma occurred ~ 0.64 Mya. In contrast, the Isolation and Migration model suggested that J. erectopatens diverged from J. microsperma due to geographic isolation during the early to middle Pleistocene about 1.24 Mya, which corresponds to the Quaternary climate oscillations. Approximate Bayesian computation (ABC) suggested that J. erectopatens underwent a population expansion ~ 726 thousand years ago (Kya) followed by a decline ~ 188 Kya. These results suggest that the Quaternary climate oscillations had a profound impact on the demographic history of the QTP endemic J. erectopatens as well as on the divergence between J. erectopatens and J. microsperma. Furthermore, they suggest that some QTP endemic woody species might be younger than previously thought.

Frankham, Greta J., Neaves, Linda E., Eldridge, Mark D. B. (2020): Genetic Relationships of Long-nosed Potoroos Potorous tridactylus (Kerr, 1792) from the Bass Strait Islands, with Notes on the Subspecies Potorous tridactylus benormi Courtney, 1963. Records of the Australian Museum 72 (5): 263-270, DOI: 10.3853/j.2201-4349.72.2020.1725, URL: http://dx.doi.org/10.3853/j.2201-4349.72.2020.1725

SummaryThe Yellow ChatEpthianura croceais comprised of three disjunct subspecies. SubspeciesE. c. macgregori(Capricorn Yellow Chat) is listed as Critically Endangered under the EPBC Act and has a distribution that also appears to be disjunct, with a limited geographic area of less than 7,000 ha. Some populations are threatened by rapid industrial development, and it is important for conservation of the subspecies to determine the extent to which the putative populations are connected. We used 14 microsatellite markers to measure genetic diversity and to determine the extent of gene flow between two disjunct populations at the northern and southern extremes of the subspecies’ range. No significant differences in genetic diversity (number of alleles and heterozygosity) were observed, but clear population structuring was apparent, with obvious differentiation between the northern and southern populations. The most likely explanation for reduced gene flow between the two populations is either the development of a geographic barrier as a consequence of shrinkage of the marine plains associated with the rise in sea levels following the last glacial maxima, or reduced connectivity across the largely unsuitable pasture and forest habitat that now separates the two populations, exacerbated by declining population size and fewer potential emigrants. Regardless of the mechanism, restricted gene flow between these two populations has important consequences for their ongoing conservation. The relative isolation of the smaller southern groups (the Fitzroy River delta and Curtis Island) from the much larger northern group (both sides of the Broad Sound) makes the southern population more vulnerable to local extinction. Conservation efforts should focus on nature refuge agreements with land owners agreeing to maintain favourable grazing management practices in perpetuity, particularly in the northern area where most chats occur. Supplemental exchanges of individuals from northern and southern populations should be explored as a way of increasing genetic diversity and reducing inbreeding.

Little genetic research has been undertaken on mammals across the vast expanse of the arid biome in Australia, despite continuing species decline and need for conservation management. Here, we evaluate the contemporary and historical genetic connectivity of the yellow-footed rock-wallaby, Petrogale xanthopus xanthopus, a threatened macropodid which inhabits rocky outcrops across the disconnected mountain range systems of the southern arid biome. We use 17 microsatellite loci together with mitochondrial control region data to determine the genetic diversity of populations and the evolutionary processes shaping contemporary population dynamics on which to base conservation recommendations. Our results indicate the highly fragmented populations have reduced diversity and limited contemporary gene flow, with most populations having been through population bottlenecks. Despite limited contemporary gene flow, the phylogeographic relationships of the mitochondrial control region indicate a lack of structure and suggests greater historical connectivity. This is an emerging outcome for mammals across this arid region. On the basis of our results, we recommend augmentation of populations of P. x. xanthopus, mixing populations from disjunct mountain range systems to reduce the chance of continued diversity loss and inbreeding depression, and therefore maximize the potential for populations to adapt and survive into the future.

The koala, the only extant species of the marsupial family Phascolarctidae, is classified as ‘vulnerable’ due to habitat loss and widespread disease. We sequenced the koala genome, producing a complete and contiguous marsupial reference genome, including centromeres. We reveal that the koala’s ability to detoxify eucalypt foliage may be due to expansions within a cytochrome P450 gene family, and its ability to smell, taste and moderate ingestion of plant secondary metabolites may be due to expansions in the vomeronasal and taste receptors. We characterized novel lactation proteins that protect young in the pouch and annotated immune genes important for response to chlamydial disease. Historical demography showed a substantial population crash coincident with the decline of Australian megafauna, while contemporary populations had biogeographic boundaries and increased inbreeding in populations affected by historic translocations. We identified genetically diverse populations that require habitat corridors and instituting of translocation programs to aid the koala’s survival in the wild., コアラの全ゲノム配列の解読に成功 --コアラはなぜ猛毒のユーカリを食べるのか？--. 京都大学プレスリリース. 2018-07-04.

Pteropus (flying-foxes) are a speciose group of non-echolocating large bats, with five extant Australian species and 24 additional species distributed amongst the Pacific Islands. In 2015, an injured flying-fox with unusual facial markings was found in Sydney, Australia, following severe and widespread storms. Based on an initial assessment, the individual belonged to Pteropus but could not be readily identified to species. As a consequence, four hypotheses for its identification/origin were posited: the specimen represented (1) an undescribed Australian species; or (2) a morphological variant of a recognised Australian species; or (3) a hybrid individual; or (4) a vagrant from the nearby Southwest Pacific Islands. We used a combination of morphological and both mitochondrial- and nuclear DNA-based identification methods to assess these hypotheses. Based on the results, we propose that this morphologically unique Pteropus most likely represents an unusual P. alecto (black flying-fox) potentially resulting from introgression from another Pteropus species. Unexpectedly, this individual, and the addition of reference sequence data from newly vouchered specimens, revealed a previously unreported P. alecto mitochondrial DNA lineage. This lineage was distinct from currently available haplotypes. It also suggests long-term hybridisation commonly occurs between P. alecto and P. conspicillatus (spectacled flying-fox). This highlights the importance of extensive reference data, and the inclusion of multiple vouchered specimens for each species to encompass both intraspecific and interspecific variation to provide accurate and robust species identification. Moreover, our additional reference data further demonstrates the complexity of Pteropus species relationships, including hybridisation, and potential intraspecific biogeographical structure that may impact on their management and conservation.

A fundamental challenge in resolving evolutionary relationships across the tree of life is to account for heterogeneity in the evolutionary signal across loci. Studies of marsupial mammals have demonstrated that this heterogeneity can be substantial, leaving considerable uncertainty in the evolutionary timescale and relationships within the group. Using simulations and a new phylogenomic data set comprising nucleotide sequences of 1550 loci from 18 of the 22 extant marsupial families, we demonstrate the power of a method for identifying clusters of loci that support different phylogenetic trees. We find two distinct clusters of loci, each providing an estimate of the species tree that matches previously proposed resolutions of the marsupial phylogeny. We also identify a well-supported placement for the enigmatic marsupial moles (Notoryctes) that contradicts previous molecular estimates but is consistent with morphological evidence. The pattern of gene-tree variation across tree-space is characterized by changes in information content, GC content, substitution-model adequacy, and signatures of purifying selection in the data. In a simulation study, we show that incomplete lineage sorting can explain the division of loci into the two tree-topology clusters, as found in our phylogenomic analysis of marsupials. We also demonstrate the potential benefits of minimizing uncertainty from phylogenetic conflict for molecular dating. Our analyses reveal that Australasian marsupials appeared in the early Paleocene, whereas the diversification of present-day families occurred primarily during the late Eocene and early Oligocene. Our methods provide an intuitive framework for improving the accuracy and precision of phylogenetic inference and molecular dating using genome-scale data.

The tammar wallaby (Notamacropus eugenii) is one of the most intensively studied of all macropodids and was the first Australasian marsupial to have its genome sequenced. However, comparatively little is known about genetic diversity and differentiation amongst the morphologically distinct allopatric populations of tammar wallabies found in Western (WA) and South Australia (SA). Here we compare autosomal and Y-linked microsatellite genotypes, as well as sequence data (~600 bp) from the mitochondrial DNA (mtDNA) control region (CR) in tammar wallabies from across its distribution. Levels of diversity at autosomal microsatellite loci were typically high in the WA mainland and Kangaroo Island (SA) populations (A = 8.9-10.6; He = 0.77-0.78) but significantly reduced in other endemic island populations (A = 3.8-4.1; He = 0.41-0.48). Autosomal and Y-linked microsatellite loci revealed a pattern of significant differentiation amongst populations, especially between SA and WA. The Kangaroo Island and introduced New Zealand population showed limited differentiation. Multiple divergent mtDNA CR haplotypes were identified within both SA and WA populations. The CR haplotypes of tammar wallabies from SA and WA show reciprocal monophyly and are highly divergent (14.5%), with levels of sequence divergence more typical of different species. Within WA tammar wallabies, island populations each have unique clusters of highly related CR haplotypes and each is most closely related to different WA mainland haplotypes. Y-linked microsatellite haplotypes show a similar pattern of divergence although levels of diversity are lower. In light of these differences, we suggest that two subspecies of tammar wallaby be recognized; Notamacropus eugenii eugenii in SA and N. eugenii derbianus in WA. The extensive neutral genetic diversity and inter-population differentiation identified within tammar wallabies should further increase the species value and usefulness as a model organism.

This study examines genetic relationships among and within the South American species of Oreobolus that span the temperate and tropical Andes hotspots and represent a good case study to investigate diversification in the Páramo. A total of 197 individuals covering the distributional range of most of these species were sequenced for the nuclear ribosomal internal transcribed spacer (ITS) and 118 individuals for three chloroplast DNA regions (trnL-F, trnH-psbA and rpl32-trnL). Haplotype networks and measures of genetic diversity were calculated at different taxonomic and geographic levels. To test for possible geographic structure, a spatial analysis of molecular variance (SAMOVA) was undertaken and species relationships were recovered using a coalescent-based approach. Results indicate complex relationships among the five South American species of Oreobolus, which are likely to have been confounded by incomplete lineage sorting, though hybridization cannot be completely discarded as an influence on genetic patterns, particularly among the northern populations of O. obtusangulus and O. cleefii. We report a case of cryptic speciation in O. obtusangulus where northern and southern populations of morphologically similar individuals are genetically distinct in all analyses. At the population level, the genetic evidence is consistent with contraction and expansion of islands of Páramo vegetation during the climatic fluctuations of the Quaternary, highlighting the role of these processes in shaping modern diversity in that ecosystem.

The rufous hare-wallaby (Lagorchestes hirsutus) is now extinct in the wild on mainland Australia, but survives in captivity. However, endemic populations persist on Bernier and Dorre Islands, Western Australia. This study aimed to compare the genetic diversity and differentiation amongst three remaining rufous hare-wallaby populations using mitochondrial DNA (mtDNA) (cytochrome b, control region) sequence data and nuclear (microsatellite) markers. Levels of microsatellite diversity were low in both island populations but high in the captive mainland population. Levels of mtDNA diversity were low in all three populations. The mainland and island populations of L. hirsutus were found to be significantly differentiated for both microsatellite and mtDNA data, but the two island populations were significantly differentiated only for the microsatellite data. This pattern of differentiation is not consistent with the recognition of two separate island subspecies, but we recommend that the mainland and island populations be regarded as separate subspecies. The low diversity of the island populations and differentiation between island and mainland populations presents both challenges and opportunities for future management.

Black-flanked rock-wallabies (Petrogale lateralis lateralis) were thought to be locally extinct in Kalbarri National Park, Western Australia, until 2015 when a pair were photographed in the Murchison River gorge. Subsequent searches failed to locate any other populations and, in combination with previous surveys, suggest that these animals were the sole survivors of a formerly abundant population. Efforts to capture the rock-wallabies to obtain tissue samples and ascertain their genetic affinities were unsuccessful. However, fresh faecal material was collected and a ~600-bp fragment of DNA sequence data was obtained from a mitochondrial DNA gene. The Kalbarri rock-wallabies were found to be genetically distinct from all other sampled populations of P. l. lateralis. They were genetically most similar to rock-wallabies in the Calvert Range, 950km to the north-east in the Little Sandy Desert. Nearer populations in Cape Range (600km north) and the Western Australian Wheatbelt (500km south-east) were less closely related. Following evaluation of a variety of factors, including population size, genetics, demography, ecology, environment and logistics, a decision was made to supplement the Kalbarri National Park population with females sourced from the Western Australian Wheatbelt. In May 2016 three females were released at the same site as the Kalbarri pair to try to ensure the survival of the population whilst still preserving unique Kalbarri alleles.

Aim: Climatic fluctuations during the Pleistocene have shaped the population structure of many extant taxa. However, few studies have examined widespread species inhabiting the Australian continent, where periods of increased aridity characterized the Pleistocene. Here we investigate the phylogeography and population history of a widespread and vagile southern Australian marsupial, the western grey kangaroo (Macropus fuliginosus). Location: Southern Australia. Methods: We examined the variation of the mitochondrial DNA (mtDNA) control region from 511 individuals of M. fuliginosus sampled throughout their transcontinental distribution. Maximum likelihood and Bayesian analyses were used to investigate the phylogeography and coalescence analyses were then used to test hypothesized biogeographical scenarios. Results: The combined results of the phylogeographical and coalescence analyses revealed a complex evolutionary history. Macropus fuliginosus originated in the south-west of the continent, with north-western and south-western populations subsequently diverging as a result of vicariance events during the mid-Pleistocene. Subsequent arid phases affected these populations differently. In the north-west, the expansion and contraction of the arid zone resulted in repeated vicariance events and multiple divergent north-western mtDNA subclades. In contrast, the south-western population was less impacted by climatic oscillations but gave rise to a major transcontinental eastward expansion. Main conclusions: Macropus fuliginosus exhibits the genetic signature of divergence due to unidentified barriers in south-western Western Australia, while previously identified barriers across southern Australia appear to have had little impact despite evidence of a broad-scale range expansion prior to the Last Glacial Maximum (LGM). This pattern of localized expansion and contraction is comparable to unglaciated regions in both the Northern and Southern Hemispheres. Furthermore, this study indicates that despite the potential similarities between Northern Hemisphere glaciation and the activation of dune systems in the Australian arid zone, both of which rendered large areas inhospitable, the biotic responses and resultant phylogeographical signatures are dissimilar. Whereas a limited number of major geographically concordant refugia are observed in glaciated areas, the Southern Hemisphere arid zone appears to be associated with multiple species-specific idiosyncratic refugia.

Threatened species often have small and isolated populations where mating among relatives can result in inbreeding depression increasing extinction risk. Effective management is hampered by a lack of syntheses summarising the magnitude of, and variation in inbreeding depression. Here we describe the nature and scope of the literature examining phenotypic/fitness consequences of inbreeding, to provide a foundation for future syntheses and management. We searched the literature for articles documenting the impact of inbreeding in natural populations. Article titles, abstracts and full-texts were assessed against a priori defined criteria, and information relating to study design, quality and other factors that may influence inbreeding responses (e.g. population size) was extracted from relevant articles. The searches identified 11457 articles, of which 614 were assessed as relevant and included in the systematic map (corresponding to 703 distinct studies). Most studies (663) assessed within-population inbreeding resulting from self-fertilisation or consanguineous pairings, while 118 studies assessed among-population inbreeding due to drift load. Plants were the most studied taxon (469 studies) followed by insects (52 studies) and birds (43 studies). Most studies investigated the effects of inbreeding on components of fitness (e.g. survival or fecundity; 648 studies) but measurements were typically under laboratory/greenhouse conditions (486 studies). Observations were also often restricted to the first inbred generation (607 studies) and studies frequently lacked contextual information (e.g. population size). Our systematic map describes the scope and quality of the evidence describing the phenotypic consequences of inbreeding. The map reveals substantial evidence relating to inbreeding responses exists, but highlights information is still limited for some aspects, including the effects of multiple generations of inbreeding. The systematic map allowed us to define several conservation-relevant questions, where sufficient data exists to support systematic reviews, e.g. How do inbreeding responses vary with population size? However, we found that such syntheses are likely to be constrained by incomplete reporting of critical contextual information. Our systematic map employed the same rigorous literature assessment methods as systematic review, including a novel survey of study quality and thus provides a robust foundation to guide future research and syntheses seeking to inform conservation decision-making.

Many endemic plant species belong to taxonomically complex groups. These endemics have often arisen as a consequence of recent and rapid evolutionary divergence facilitated by processes such as hybridization, polyploidy and/or breeding system transitions. The rapid and dynamic nature of divergence in taxonomically complex groups leads to problems in the implementation of traditional species-based approaches for the conservation of the biodiversity that they contain. Firstly, the taxa of interest can be difficult to define and identify, leading to practical difficulties in implementing conservation measures. Secondly, a species-based approach often fails to capture the complexity of diversity present in the taxonomically complex group. To accommodate these challenges, we have developed a Process-Based Species Action Plan approach. This is designed to conserve the processes leading to the generation of biodiversity, rather than focusing on the preservation of individual named taxa. We illustrate the approach using a group of endemic tree species (Sorbus) on the Scottish island of Arran that have originated via a combination of multiple recent hybridization events and apomixis. The plan focuses on the optimization of habitat management to ensure the reproduction and regeneration of Sorbus in the zone in which these evolutionary processes operate, and to facilitate hybridization that will ensure the continued generation of diversity in this group. © 2011 The Linnean Society of London, Botanical Journal of the Linnean Society, 2012, 168, 194–203.

The brush-tailed rock-wallaby (Petrogale penicillata) is now extinct west of the Great Dividing Range, apart from remnant populations in the Warrumbungles and at Mt Kaputar. Previous genetic analysis has identified deep genetic subdivisions within P. penicillata, but samples from Mt Kaputar were not included. Mitochondrial DNA sequences obtained from Mt Kaputar and the Warrumbungles clustered within the Central ESU, extending its distribution north of the Hunter River onto the north-western slopes adjacent to populations of the Northern ESU. These highly threatened western populations are the only P. penicillata persisting in semiarid conditions. This makes them of particular value to the long-term survival of the species and their conservation should be a priority.

Introgressive hybridization has traditionally been regarded as rare in many vertebrate groups, including mammals. Despite a propensity to hybridize in captivity, introgression has rarely been reported between wild sympatric macropodid marsupials. Here we investigate sympatric populations of western (Macropus fuliginosus) and eastern (Macropus giganteus) grey kangaroos through 12 autosomal microsatellite loci and 626 bp of the hypervariable mitochondrial DNA (mtDNA) control region. M. fuliginosus and M. giganteus within the region of sympatry corresponded, both genetically and morphologically, to their respective species elsewhere in their distributions. Of the 223 grey kangaroos examined, 7.6% displayed evidence of introgression, although no F1 hybrids were detected. In contrast to captive studies, there was no evidence for unidirectional hybridization in sympatric grey kangaroos. However, a higher portion of M. giganteus backcrosses existed within the sample compared with M. fuliginosus. Hybridization in grey kangaroos is reflective of occasional breakdowns in species boundaries, occurring throughout the region and potentially associated with variable conditions and dramatic reductions in densities. Such rare hybridization events allow populations to incorporate novel diversity while still retaining species integrity.

Animals exhibit a range of dispersal strategies that impact on the organisation of individuals and can be influenced by both the environment and population demography. We examined the fine-scale spatial genetic structure and patterns of relatedness in 139 adult eastern grey kangaroos (Macropus giganteus) to test predictions of male-biased dispersal and female philopatry in comparison with previous studies in different environments in the species’ distributions. We found evidence of limited differences between the sexes, with little spatial genetic structure in both males and females. The levels of relatedness among females in close proximity were not indicative of close relatives (e.g. mother–daughter) and there was no evidence of matrilineal structure. Among males, there was little evidence of genetic structure. Although our results are, in general, consistent with those of previous studies, we found study-specific differences in the extent of genetic structure that appear to be related to differences in environmental and demographic conditions across the distribution. This highlights the need for additional research focussing on populations from a range of environmental conditions.

Planting of native trees for habitat restoration is a widespread practice, but the consequences for the retention and transmission of genetic diversity in planted and natural populations are unclear. Using Inga vera subsp. affinis as a model species, we genotyped five natural and five planted populations in the Atlantic forest of northeastern Brazil at polymorphic microsatellite loci. We studied the breeding system and population structure to test how much genetic diversity is retained in planted relative to natural populations. We then genotyped seedlings from these populations to test whether genetic diversity in planted populations is restored by outcrossing to natural populations of I. vera. The breeding system of natural I. vera populations was confirmed to be highly outcrossing (t = 0.92; FIS = 20.061, P = 0.04), with populations showing weak population substructure (FST = 0.028). Genetic diversity in planted populations was 50% less than that of natural populations (planted: AR = 14.9, HO = 0.865 and natural: AR = 30.8, HO = 0.655). However, seedlings from planted populations showed a 30% higher allelic richness relative to their parents (seedlings AR = 10.5, parents AR = 7.6). Understanding the processes and interactions that shape this system are necessary to provide ecologically sensible goals and successfully restore hyperfragmented habitats. Future restoration plans for I. vera must consider the genetic diversity of planted populations and the potential for gene flow between natural populations in the landscape, in order to preserve ecological interactions (i.e. pollination), and promote opportunities for outcrossing.

Habitat loss and fragmentation are key threats to local koala (Phascolarctos cinereus) populations. Broad-scale management is suboptimal for koalas because distribution models are not easily generalised across regions. Therefore, it is imperative that data relevant to local management bodies are available. Genetic data provides important information on gene flow and potential habitat barriers, including anthropogenic disturbances. Little genetic data are available for nationally significant koala populations in north-eastern New South Wales, despite reported declines due to urbanisation and habitat loss. In this study, we develop 14 novel microsatellite loci to investigate koala populations in north-eastern New South Wales (Port Macquarie, Coffs Harbour, Tyagarah, Ballina) and south-eastern Queensland (Coomera). All locations were significantly differentiated (FST = 0.096–0.213; FʹST = 0.282–0.582), and this pattern was not consistent with isolation by distance (R2 = 0.228, P = 0.058). Population assignment clustered the more northern populations (Ballina, Tyagarah and Coomera), suggesting contemporary gene flow among these sites. For all locations, low molecular variation among (16%) rather than within (84%) sites suggests historical connectivity. These results suggest that koala populations in north-eastern New South Wales and south-eastern Queensland are experiencing contemporary impediments to gene flow, and highlight the importance of maintaining habitat connectivity across this region.

Large vagile mammals typically exhibit little genetic structuring across their range, particularly when their habitat is essentially continuous. We investigated the population genetic structure of a large vagile Australian macropodid, Macropus fuliginosus, which is continuously distributed across most of southern Australia, using nine highly polymorphic nuclear microsatellite loci. Five distinct genetic units were identified across the range, four on the mainland and one on Kangaroo Island. In addition to the predicted historic Nullarbor Plain Barrier, two unexpected mainland barriers to gene flow were identified. Both were associated with landscape discontinuities (Swan River, Flinders Ranges), which appear within the dispersal capabilities of M. fuliginosus. Typical of large vagile mammals, M. fuliginosus displays high genetic diversity (with the exception of an insular population) and weak genetic structuring (within genetic units). However, the expansion of M. fuliginosus from southwestern Australia during the Pleistocene has resulted in significantly reduced genetic diversity in eastern populations. No significant sex-biased dispersal was detected, although differences in habitat, densities and climatic conditions between the eastern and western regions of the range appear to influence dispersal with the effects of isolation by distance only evident in the west. These results suggest that the biogeography of southern Australia is more complex than previously thought and reveal that seemingly minor landscape features can significantly impact genetic structuring in large vagile mammals.

The majority of research into the timing of gonad differentiation (and sex determination) in reptiles has focused on oviparous species. This is largely because: (1) most reptiles are oviparous; (2) it is easier to manipulate embryonic developmental conditions (e.g., temperature) of eggs than oviductal embryos and (3) modes of sex determination in oviparous taxa were thought to be more diverse since viviparity and environmental sex determination (ESD)/temperature-dependent sex determination (TSD) were considered incompatible. However, recent evidence suggests the two may well be compatible biological attributes, opening potential new lines of enquiry into the evolution and maintenance of sex determination. Unfortunately, the baseline information on embryonic development in viviparous species is lacking and information on gonad differentiation and sexual organ development is almost non-existent. Here we present an embryonic morphological development table (10 stages), the sequence of gonad differentiation and sexual organ development for the viviparous spotted snow skink (Niveoscincus ocellatus). Gonad differentiation in this species is similar to other reptilian species. Initially, the gonads are indifferent and both male and female accessory ducts are present. During stage 2, in the middle third of development, differentiation begins as the inner medulla regresses and the cortex thickens signaling ovary development, while the opposite occurs in testis formation. At this point, the Mullerian (female reproductive) duct regresses in males until it is lost (stage 6), while females retain both ducts until after birth. In the later stages of testis development, interstitial tissue forms in the medulla corresponding to maximum development of the hemipenes in males and the corresponding regression in the females.

There are several aspects of biology in which the contribution of males and females is unequal. In these instances the examination of Y chromosome markers may be used to elucidate male-specific attributes. Here, male dispersal patterns and genetic structuring were examined using four Y-microsatellite loci in 186 male western grey kangaroos, Macropus fuliginosus, from throughout the species’ trans-continental distribution. In addition, 52 male grey kangaroos were examined to investigate hybridisation between M. fuliginosus and the eastern grey kangaroo, Macropus giganteus, in their region of sympatry in eastern Australia. Detected Y chromosome diversity was low, resulting from low effective male population size due to skewed sex ratios and a polygynous mating system. As expected, male dispersal was high across the range. However, the Lake Torrens–Flinders Ranges region appears to have significantly restricted male movement between eastern and central/western Australia. There was little evidence to suggest that other barriers (Nullarbor Plain and Swan River Valley) previously identified by nuclear and mitochondrial DNA marker studies restrict male movement. Hence, the admixture events previously identified may be associated with high male dispersal. Within the region of sympatry between M. fuliginosus and M. giganteus in eastern Australia, four M. giganteus individuals were found to possess M. fuliginosus Y-haplotypes. These results confirm the occurrence of hybridisation between male M. fuliginosus and female M. giganteus. Additionally, the introgression of M. fuliginosus Y-haplotypes into M. giganteus populations indicates that at least some male hybrids are fertile, despite evidence to the contrary from captive studies. This study has provided insights into the male contribution to population history, structure and hybridisation in M. fuliginosus, which were not predicted by comparisons between biparentally and maternally inherited markers. This highlights the importance of direct examination of the Y chromosome to provide novel insights into male-mediated processes, especially where the contribution of the sexes may differ.

Although pademelons (Thylogale) are widespread and common in coastal eastern Australia, they have been largely neglected in population genetic studies. Here we use 10 microsatellite loci and the mitochondrial DNA (mtDNA) cytochrome b (Cytb) gene to examine genetic differentiation amongst populations of the red-necked (Thylogale thetis) and red-legged (T. stigmatica) pademelon in eastern Australia. Evidence of hybridisation was detected between subspecies of T. stigmatica in central Queensland. Specimens sampled between Eungella and Sarina were found to represent a broad (~90 km) zone of introgression, for both nuclear and mtDNA markers, between T. s. stigmatica (Wet Tropics) and T. s. wilcoxi (south-east Queensland). In addition, individuals sampled from around Proserpine were genetically T. s. stigmatica rather than T. s. wilcoxi, as had previously been assumed. This observation raises some intriguing questions about the dispersal ability of T. stigmatica and the phylogeographic history of moist forest taxa in eastern Australia. Only limited evidence of introgression was detected between sympatric populations of T. thetis and T. s. wilcoxi in south-east Queensland.

Road-killed animals are a readily available but often overlooked source of specimens and data. Determining the sex of road-killed possums is often difficult due to the severe damage carcasses sustain on impact with vehicles. Consequently, valuable information is lost. To assess the potential application of genetic techniques to address this problem, tissue biopsies (n = 47) taken from road-killed possums were sexed via PCR, through the simultaneous use of W- (G6PD) and Y-linked (SRY) primers. Comparisons with animals of known sex were used to validate the accuracy of this method, with 100% (n = 19) of samples correctly identified in a blind test. This methodology can provide researchers with valuable, additional data for road-kill studies where assigning sex in the field is not possible.

The common brushtail possum (Trichosurus vulpecula) is usually a strictly monotocous species, presenting one offspring at a time. Two concurrent offspring is a rare occurrence, but twin pouch young or young spaced one oestrous cycle apart have been reported. We investigated the biology of urban T. vulpecula populations within the Sydney metropolitan area between November 2002 and April 2005. As part of these studies, we trapped one adult female T.�vulpecula with two young. Analysis of morphological data from the young and development of the female?s mammary gland suggest that only one young was her offspring. The results of genetic analysis support this conclusion. Herewith, we describe an apparent case of adoption in T. vulpecula.

The Australian continent exhibits complex biogeographic patterns but studies of the impacts of Pleistocene climatic oscillation on the mesic environments of the Southern Hemisphere are limited. The koala (Phascolarctos cinereus), one of Australia’s most iconic species, was historically widely distributed throughout much of eastern Australia but currently represents a complex conservation challenge. To better understand the challenges to koala genetic health, we assessed the phylogeographic history of the koala. Variation in the maternally inherited mitochondrial DNA (mtDNA) Control Region (CR) was examined in 662 koalas sampled throughout their distribution. In addition, koala CR haplotypes accessioned to Genbank were evaluated and consolidated. A total of 53 unique CR haplotypes have been isolated from koalas to date (including 15 haplotypes novel to this study). The relationships among koala CR haplotypes were indicative of a single Evolutionary Significant Unit and do not support the recognition of subspecies, but were separated into four weakly differentiated lineages which correspond to three geographic clusters: a central lineage, a southern lineage and two northern lineages co-occurring north of Brisbane. The three geographic clusters were separated by known Pleistocene biogeographic barriers: the Brisbane River Valley and Clarence River Valley, although there was evidence of mixing amongst clusters. While there is evidence for historical connectivity, current koala populations exhibit greater structure, suggesting habitat fragmentation may have restricted female-mediated gene flow. Since mtDNA data informs conservation planning, we provide a summary of existing CR haplotypes, standardise nomenclature and make recommendations for future studies to harmonise existing datasets. This holistic approach is critical to ensuring management is effective and small scale local population studies can be integrated into a wider species context.