Location, location, location: survival of Antarctic biota requires the best real estate.

The origin of terrestrial biota in Antarctica has been debated since the discovery of springtails on the first historic voyages to the southern continent more than 120 years ago. A plausible explanation for the long-term persistence of life requiring ice-free land on continental Antarctica has, however, remained elusive. The default glacial eradication scenario has dominated because hypotheses to date have failed to provide a mechanism for their widespread survival on the continent, particularly through the Last Glacial Maximum when geological evidence demonstrates that the ice sheet was more extensive than present. Here, we provide support for the alternative nunatak refuge hypothesis-that ice-free terrain with sufficient relief above the ice sheet provided refuges and was a source for terrestrial biota found today. This hypothesis is supported here by an increased understanding from the combination of biological and geological evidence, and we outline a mechanism for these refuges during successive glacial maxima that also provides a source for coastal species. Our cross-disciplinary approach provides future directions to further test this hypothesis that will lead to new insights into the evolution of Antarctic landscapes and how they have shaped the biota through a changing climate.

The presence of a guard vicariously drives split sex ratios in a facultatively social bee.

Split sex ratios provide broad insights into how reproductive strategies evolve, and historically have special relevance to the evolution of eusociality. Yet almost no attention has been directed to situations where split sex ratios may potentially decrease the payoffs for worker-like behaviour, increasing selective thresholds for eusociality. We examined sex ratios in a facultatively social colletid bee, Amphylaeus morosus. Sex ratios in this bee vary strongly with the presence of a nest guard and in a pattern that does not conform to assumptions of previous models in which split sex ratios facilitate altruism. While the production of daughters was constant across social and solitary nests, mothers produced more brood when a non-reproductive guard was present, but these extra brood were all male. This leads to split sex ratios, vicariously driven by guards that are unable to manipulate sex ratios in their favour. Importantly, if guarding becomes more common in a population this would lead to an excess of males and lower the genetic value of these extra males to guards, effectively putting a brake on selection for worker-like behaviour.

Holocene population expansion of a tropical bee coincides with early human colonization of Fiji rather than climate change.

There is substantial debate about the relative roles of climate change and human activities on biodiversity and species demographies over the Holocene. In some cases, these two factors can be resolved using fossil data, but for many taxa such data are not available. Inferring historical demographies of taxa has become common, but the methodologies are mostly recent and their shortcomings often unexplored. The bee genus Homalictus is developing into a tractable model system for understanding how native bee populations in tropical islands have responded to past climate change. We greatly expand on previous studies using sequences of the mitochondrial gene COI from 474 specimens and between 171 and 3928 autosomal (DArTSeq) single nucleotide polymorphism loci from 19 specimens of the native Fijian bee, Homalictus fijiensis, to explore its historical demography using coalescent and mismatch analyses. We ask whether past changes in demography were human- or climate-driven, while considering analytical assumptions. We show that inferred changes in population sizes are too recent to be explained by past climate change. Instead we find that a dramatic increase in population size for the main island of Viti Levu coincides with increasing occupation by humans and their modification of the environment. We found no corresponding change in bee population size for another major island, Kadavu, where human populations and agricultural activities have been historically very low. Our analyses indicate that molecular approaches can be used to disentangle the impacts of humans and climate change on a major tropical pollinator and that stringent analytical approaches are required for reliable interpretation of results.

Climate change and invasive species: a physiological performance comparison of invasive and endemic bees in Fiji.

Anthropogenic climate change and invasive species are two of the greatest threats to biodiversity, affecting the survival, fitness and distribution of many species around the globe. Invasive species are often expected to have broad thermal tolerance, be highly plastic, or have high adaptive potential when faced with novel environments. Tropical island ectotherms are expected to be vulnerable to climate change as they often have narrow thermal tolerance and limited plasticity. In Fiji, only one species of endemic bee, Homalictus fijiensis, is commonly found in the lowland regions, but two invasive bee species, Braunsapis puangensis and Ceratina dentipes, have recently been introduced into Fiji. These introduced species pollinate invasive plants and might compete with H. fijiensis and other native pollinators for resources. To test whether certain performance traits promote invasiveness of some species, and to determine which species are the most vulnerable to climate change, we compared the thermal tolerance, desiccation resistance, metabolic rate and seasonal performance adjustments of endemic and invasive bees in Fiji. The two invasive species tended to be more resistant to thermal and desiccation stress than H. fijiensis, while H. fijiensis had greater capacity to adjust their CTmax with season, and H. fijiensis females tended to have higher metabolic rates than B. puangensis females. These findings provide mixed support for current hypotheses for the functional basis of the success of invasive species; however, we expect the invasive bees in Fiji to be more resilient to climate change because of their increased thermal tolerance and desiccation resistance.

Molecular diversity and species delimitation in the family Gasteruptiidae (Hymenoptera: Evanioidea).

Gasteruptiidae Ashmead is an easily recognised family of wasps with ∼589 described species worldwide. Although well characterised by traditional taxonomy, multiple authors have commented on the extreme morphological uniformity of the group, making species-level identification difficult. This problem is enhanced by the lack of molecular data and molecular phylogenetic research for the group. We used 187 cytochrome c oxidase subunit I (COI) barcodes to explore the efficiency of sequence data to delimitate species in Gasteruptiidae. We undertook a graphical and discussion-based comparison of six methods for species delimitation, with the success of methods judged based on known species boundaries and morphology. Both distance-based (ABGD and jMOTU threshold analysis) and tree-based (GMYC and PTP) methods compared across multiple parameters recovered variable molecular operational taxonomic units (MOTUs), ranging from 55 to 123 MOTUs. Tree-based methods tended to split known morphological species less than distance-based methods, with the single-threshold GMYC method the most concordant with known morphospecies. Our results suggest that the incorporation of molecular species delimitation techniques provides a powerful tool to assist in the interpretation of species and help direct informed decisions with taxonomic uncertainty in the family.

Extreme genetic diversity among springtails (Collembola) in subterranean calcretes of arid Australia.

The subterranean islands hypothesis for calcretes of the Yilgarn region in Western Australia applies to many stygobitic (subterranean-aquatic) species that are "trapped" evolutionarily within isolated aquifers due to their aquatic lifestyles. In contrast, little is known about the distribution of terrestrial-subterranean invertebrates associated with the calcretes. We used subterranean Collembola from the Yilgarn calcretes to test the hypothesis that troglobitic species, those inhabiting the subterranean unsaturated (non-aquatic) zone of calcretes, are also restricted in their distribution and represent reciprocally monophyletic and endemic lineages. We used the barcoding fragment of the mtDNA cytochrome c oxidase subunit 1 (COI) gene from 183 individuals to reconstruct the phylogenetic history of the genus Pseudosinella Schäffer (Collembola, Lepidocyrtidae) from 10 calcretes in the Yilgarn. These calcretes represent less than 5% of the total possible calcretes in this region, yet we show that their diversity for subterranean Collembola comprises a minimum of 25 new species. Regionally, multiple levels of diversity exist in Pseudosinella, indicative of a complex evolutionary history for this genus in the Yilgarn. These species have probably been impacted by climatic oscillations, facilitating their dispersal across the landscape. The results represent a small proportion of the undiscovered diversity in Australia's arid zone.

Radiation of tropical island bees and the role of phylogenetic niche conservatism as an important driver of biodiversity.

Island biogeography explores how biodiversity in island ecosystems arises and is maintained. The topographical complexity of islands can drive speciation by providing a diversity of niches that promote adaptive radiation and speciation. However, recent studies have argued that phylogenetic niche conservatism, combined with topographical complexity and climate change, could also promote speciation if populations are episodically fragmented into climate refugia that enable allopatric speciation. Adaptive radiation and phylogenetic niche conservatism therefore both predict that topographical complexity should encourage speciation, but they differ strongly in their inferred mechanisms. Using genetic (mitochondrial DNA (mtDNA) and single-nucleotide polymorphism (SNP)) and morphological data, we show high species diversity (22 species) in an endemic clade of Fijian Homalictus bees, with most species restricted to highlands and frequently exhibiting narrow geographical ranges. Our results indicate that elevational niches have been conserved across most speciation events, contradicting expectations from an adaptive radiation model but concordant with phylogenetic niche conservatism. Climate cycles, topographical complexity, and niche conservatism could interact to shape island biodiversity. We argue that phylogenetic niche conservatism is an important driver of tropical island bee biodiversity but that this phylogenetic inertia also leads to major extinction risks for tropical ectotherms under future warming climates.

Estimating the biodiversity of terrestrial invertebrates on a forested island using DNA barcodes and metabarcoding data.

Invertebrates are a major component of terrestrial ecosystems, however, estimating their biodiversity is challenging. We compiled an inventory of invertebrate biodiversity along an elevation gradient on the temperate forested island of Hauturu, New Zealand, by DNA barcoding of specimens obtained from leaf litter samples and pitfall traps. We compared the barcodes and biodiversity estimates from this data set with those from a parallel DNA metabarcoding analysis of soil from the same locations, and with pre-existing sequences in reference databases, before exploring the use of combined data sets as a basis for estimating total invertebrate biodiversity. We obtained 1,282 28S and 1,610 COI barcodes from a total of 1,947 invertebrate specimens, which were clustered into 247 (28S) and 366 (COI) OTUs, of which ≤ 10% were represented in GenBank. Coleoptera were most abundant (730 sequenced specimens), followed by Hymenoptera, Diptera, Lepidoptera, and Amphipoda. The most abundant OTU from both the 28S (153 sequences) and COI (140 sequences) data sets was an undescribed beetle from the family Salpingidae. Based on the occurrences of COI OTUs along the elevation gradient, we estimated there are ~1,000 arthropod species (excluding mites) on Hauturu, including 770 insects, of which 344 are beetles. A DNA metabarcoding analysis of soil DNA from the same sites resulted in the identification of similar numbers of OTUs in most invertebrate groups compared with the DNA barcoding, but less than 10% of the DNA barcoding COI OTUs were also detected by the metabarcoding analysis of soil DNA. A mark-recapture analysis based on the overlap between these data sets estimated the presence of approximately 6,800 arthropod species (excluding mites) on the island, including ~3,900 insects. Estimates of New Zealand-wide biodiversity for selected arthropod groups based on matching of the COI DNA barcodes with pre-existing reference sequences suggested over 13,200 insect species are present, including 4,000 Coleoptera, 2,200 Diptera, and 2,700 Hymenoptera species, and 1,000 arachnid species (excluding mites). These results confirm that metabarcoding analyses of soil DNA tends to recover different components of terrestrial invertebrate biodiversity compared to traditional invertebrate sampling, but the combined methods provide a novel basis for estimating invertebrate biodiversity.

Analysis of the genome of the New Zealand giant collembolan (Holacanthella duospinosa) sheds light on hexapod evolution.

BACKGROUND: The New Zealand collembolan genus Holacanthella contains the largest species of springtails (Collembola) in the world. Using Illumina technology we have sequenced and assembled a draft genome and transcriptome from Holacanthella duospinosa (Salmon). We have used this annotated assembly to investigate the genetic basis of a range of traits critical to the evolution of the Hexapoda, the phylogenetic position of H. duospinosa and potential horizontal gene transfer events. RESULTS: Our genome assembly was ~375 Mbp in size with a scaffold N50 of ~230 Kbp and sequencing coverage of ~180×. DNA elements, LTRs and simple repeats and LINEs formed the largest components and SINEs were very rare. Phylogenomics (370,877 amino acids) placed H. duospinosa within the Neanuridae. We recovered orthologs of the conserved sex determination genes thought to play a role in sex determination. Analysis of CpG content suggested the absence of DNA methylation, and consistent with this we were unable to detect orthologs of the DNA methyltransferase enzymes. The small subunit rRNA gene contained a possible retrotransposon. The Hox gene complex was broken over two scaffolds. For chemosensory ability, at least 15 and 18 ionotropic glutamate and gustatory receptors were identified, respectively. However, we were unable to identify any odorant receptors or their obligate co-receptor Orco. Twenty-three chitinase-like genes were identified from the assembly. Members of this multigene family may play roles in the digestion of fungal cell walls, a common food source for these saproxylic organisms. We also detected 59 and 96 genes that blasted to bacteria and fungi, respectively, but were located on scaffolds that otherwise contained arthropod genes. CONCLUSIONS: The genome of H. duospinosa contains some unusual features including a Hox complex broken over two scaffolds, in a different manner to other arthropod species, a lack of odorant receptor genes and an apparent lack of environmentally responsive DNA methylation, unlike many other arthropods. Our detection of candidate horizontal gene transfer candidates confirms that this phenomenon is occurring across Collembola. These findings allow us to narrow down the regions of the arthropod phylogeny where key innovations have occurred that have facilitated the evolutionary success of Hexapoda.

Molecular systematics and biodiversity of oniscidean isopods in the groundwater calcretes of central Western Australia.

Groundwater calcrete aquifers of central Western Australia have been shown to contain a high diversity of stygobiont (subterranean aquatic) invertebrates, with each species confined to an individual calcrete and the entire system resembling a 'subterranean archipelago' containing hundreds of isolated calcretes. Here, we utilised alternative sampling techniques above the water table and uncovered a significant fauna of subterranean terrestrial oniscidean isopods from the calcretes. We explored the diversity and evolution of this fauna using molecular analyses based on one mitochondrial gene, Cytochrome C Oxidase Subunit I (COI), two Ribosomal RNA genes (28S and 18S), and one protein coding nuclear gene, Lysyl-tRNA Synthetase (LysRS). The results from 12 calcretes showed the existence of 36 divergent DNA lineages belonging to four oniscidean families (Paraplatyarthridae, Armadillidae, Stenoniscidae and Philosciidae). Using a combination of phylogenetic and species delimitation methods, we hypothesized the occurrence of at least 27 putative new species of subterranean oniscideans, of which 24 taxa appeared to be restricted to an individual calcrete, lending further support to the "subterranean island hypothesis". Three paraplatyarthrid species were present on adjacent calcretes and these exceptions possessed more ommatidia and body pigments compared with the calcrete-restricted taxa, and are likely to represent troglophiles. The occurrence of stenoniscid isopods in the calcretes of central Western Australia, a group previously only known from the marine littoral zone, suggests a link to the marine inundation of the Eucla basin during the Late Eocene. The current oniscidean subterranean fauna consists of groups known to be subtropical, littoral and benthic, reflecting different historical events that have shaped the evolution of the fauna in the calcretes.

Parallel responses of bees to Pleistocene climate change in three isolated archipelagos of the southwestern Pacific.

The impacts of glacial cycles on the geographical distribution and size of populations have been explored for numerous terrestrial and marine taxa. However, most studies have focused on high latitudes, with only a few focused on the response of biota to the last glacial maximum (LGM) in equatorial regions. Here, we examine how population sizes of key bee fauna in the southwest Pacific archipelagos of Fiji, Vanuatu and Samoa have fluctuated over the Quaternary. We show that all three island faunas suffered massive population declines, roughly corresponding in time to the LGM, followed by rapid expansion post-LGM. Our data therefore suggest that Pleistocene climate change has had major impacts across a very broad tropical region. While other studies indicate widespread Holarctic effects of the LGM, our data suggest a much wider range of latitudes, extending to the tropics, where these climate change repercussions were important. As key pollinators, the inferred changes in these bee faunas may have been critical in the development of the diverse Pacific island flora. The magnitude of these responses indicates future climate change scenarios may have alarming consequences for Pacific island systems involving pollinator-dependent plant communities and agricultural crops.

Molecular phylogeny reveals independent origins of body scales in Entomobryidae (Hexapoda: Collembola).

Entomobryidae is the largest family in Collembola but relationships within the family have never been subjected to rigorous phylogenetic analyses. Within the family, body scales are present in many species, and are fundamental in the classification at the subfamilial and tribal levels. A molecular phylogeny was reconstructed using the nuclear 18SrRNA and partial 28SrRNA and the mitochondrial 16SrRNA to examine the evolution of scales across Entomobryidae subfamilies. These datasets were analyzed separately and combined, with parsimony, likelihood and Bayesian algorithms. Monophyly of Orchesellinae was not recovered, and it was split into a scaled clade and an unscaled clade, contradicting to all recent taxonomic conceptions. The monophyly of Entomobryinae, Seirinae and Lepidocyrtinae is well supported however within Entomobryinae, the polyphyly of Entomobryini and Willowsiini implies that classification using the presence/absence of scales is not valid. Analyses of ancestral character state reconstruction in Entomobryidae indicate that the presence of body scales have evolved independently at least five times, with a loss of scales occurring independently at least twice. A revision of the family Entomobryidae on molecular and morphological basis is clearly needed.

Biogeographic survey of soil bacterial communities across Antarctica.

BACKGROUND: Antarctica and its unique biodiversity are increasingly at risk from the effects of global climate change and other human influences. A significant recent element underpinning strategies for Antarctic conservation has been the development of a system of Antarctic Conservation Biogeographic Regions (ACBRs). The datasets supporting this classification are, however, dominated by eukaryotic taxa, with contributions from the bacterial domain restricted to Actinomycetota and Cyanobacteriota. Nevertheless, the ice-free areas of the Antarctic continent and the sub-Antarctic islands are dominated in terms of diversity by bacteria. Our study aims to generate a comprehensive phylogenetic dataset of Antarctic bacteria with wide geographical coverage on the continent and sub-Antarctic islands, to investigate whether bacterial diversity and distribution is reflected in the current ACBRs. RESULTS: Soil bacterial diversity and community composition did not fully conform with the ACBR classification. Although 19% of the variability was explained by this classification, the largest differences in bacterial community composition were between the broader continental and maritime Antarctic regions, where a degree of structural overlapping within continental and maritime bacterial communities was apparent, not fully reflecting the division into separate ACBRs. Strong divergence in soil bacterial community composition was also apparent between the Antarctic/sub-Antarctic islands and the Antarctic mainland. Bacterial communities were partially shaped by bioclimatic conditions, with 28% of dominant genera showing habitat preferences connected to at least one of the bioclimatic variables included in our analyses. These genera were also reported as indicator taxa for the ACBRs. CONCLUSIONS: Overall, our data indicate that the current ACBR subdivision of the Antarctic continent does not fully reflect bacterial distribution and diversity in Antarctica. We observed considerable overlap in the structure of soil bacterial communities within the maritime Antarctic region and within the continental Antarctic region. Our results also suggest that bacterial communities might be impacted by regional climatic and other environmental changes. The dataset developed in this study provides a comprehensive baseline that will provide a valuable tool for biodiversity conservation efforts on the continent. Further studies are clearly required, and we emphasize the need for more extensive campaigns to systematically sample and characterize Antarctic and sub-Antarctic soil microbial communities. Video Abstract.

Rapid diversification in Australia and two dispersals out of Australia in the globally distributed bee genus, Hylaeus (Colletidae: Hylaeinae).

Hylaeus is the only globally distributed colletid bee genus, with subgeneric and species-level diversity highest in Australia. We used one mitochondrial and two nuclear genes to reconstruct a phylogeny using Bayesian analyses of this genus based on species from Australia, Asia, Africa, Europe, Hawai'i, the New World and New Zealand. Our results concord with a ca. 30Mya Hylaeus crown age inferred by earlier studies, and we show that Hylaeus originated in Australia. Our phylogeny indicates only two dispersal events out of Australia, both shortly after the initial diversification of extant taxa. One of these dispersals was into New Zealand with only a minor subsequent radiation, but the second dispersal out of Australia resulted in a world-wide distribution. This second dispersal and radiation event, combined with very extensive early radiation of Hyleaus in Australia, poses a conundrum: what kinds of biogeographical and ecological factors could simultaneously drive global dispersal, yet strongly constrain further successful migrations out of Australia when geographical barriers appear to be weak? We argue that for hylaeine bees movement into new niches and enemy-free spaces may have favoured initial dispersal events, but that subsequent dispersals would not have entailed the original benefits of new niche space.

Diversification of Fijian halictine bees: insights into a recent island radiation.

Although bees form a key pollinator suite for flowering plants, very few studies have examined the evolutionary radiation of non-domesticated bees over human time-scales. This is surprising given the importance of bees for crop pollination and the effect of humans in transforming ecosystems via agriculture. In the Pacific, where the bee fauna appears depauperate, their importance as pollinators is not clear, particularly in Fiji where species diversity is even lower than neighbouring archipelagos. Here we explore the radiation of halictine bees in Fiji using phylogenetic analyses of mtDNA COI sequence data. Our analyses indicate the existence of several 'deep' clades whose divergences are close to the crown node, along with a highly derived 'broom' clade showing very high haplotype diversity, and mostly limited to low-lying agricultural regions. This derived clade is very abundant, whereas the more basal clades were relatively rare. Although nearly all haplotype diversity in Fijian Homalictus comprises synonymous substitutions, a small number of amino acid changes are associated with the major clades, including the hyper-diverse clade. Analyses of haplotype lineage accumulation show a steep increase in selectively neutral COI haplotypes corresponding to the emergence of this 'broom' clade. We explore three possible scenarios for this dramatic increase: (i) a key change in adaptedness to the environment, (ii) a large-scale extinction event, or (iii) a dramatic increase in suitable habitats leading to rapid population expansion. Using estimated mutation rates of mitochondrial DNA in other invertebrates, we argue that Homalictus first colonised the Fijian archipelago in the middle-late Pleistocene, and the rapid accumulation of haplotypes in the hyper-diverse clade occurred in the Holocene, but prior to recorded human presence in the Fijian region. Our results indicate that bees have not been important pollinators of Fijian ecosystems until very recent times. Post-Pleistocene climate change and anthropogenic effects on Fijian ecosystems are likely to have greatly transformed pollinator suites from the conditions when those ecosystems were first being assembled.

A new species of the blind cave gudgeon Milyeringa (Pisces: Gobioidei, Eleotridae) from Barrow Island, Western Australia, with a redescription of M. veritas Whitley.

A new species of the eyeless eleotrid genus Milyeringa is described from wells sunk on Barrow Island, Western Australia. Milyeringa justitia n. sp. is the third species of the genus to be named. Morphological data and cytochrome c oxidase subunit I (COI) DNA sequence data from a wide sample of localities at which the genus occurs was used to evaluate relationships and species limits. Milyeringa veritas is redescribed, and M. brooksi is synonymised with M. veritas. The unique form and ecology of these fishes, plus the threats to their survival, warrants immediate and continuing attention in management.

The use of different 16S rRNA gene variable regions in biogeographical studies.

16S rRNA gene amplicon sequencing is routinely used in environmental surveys to identify microbial diversity and composition of the samples of interest. The dominant sequencing technology of the past decade (Illumina) is based on the sequencing of 16S rRNA hypervariable regions. Online sequence data repositories, which represent an invaluable resource for investigating microbial distributional patterns across spatial, environmental or temporal scales, contain amplicon datasets from diverse 16S rRNA gene variable regions. However, the utility of these sequence datasets is potentially reduced by the use of different 16S rRNA gene amplified regions. By comparing 10 Antarctic soil samples sequenced for five different 16S rRNA amplicons, we explore whether sequence data derived from diverse 16S rRNA variable regions can be validly used as a resource for biogeographical studies. Patterns of shared and unique taxa differed among samples as a result of variable taxonomic resolutions of the assessed 16S rRNA variable regions. However, our analyses also suggest that the use of multi-primer datasets for biogeographical studies of the domain Bacteria is a valid approach to explore bacterial biogeographical patterns due to the preservation of bacterial taxonomic and diversity patterns across different variable region datasets. We deem composite datasets useful for biogeographical studies.

A taxonomic revision of the tube-web spiders of the genus Ariadna (Araneae: Segestriidae) in Tasmania.

The genus Ariadna Audouin, 1826 is revised for Tasmania to include 13 species, ten of which are described as new: Ariadna abbreviata sp. nov., A. alta sp. nov., A. amabilia sp. nov., A. crypticola sp. nov., A. ferrogrisea sp. nov., A. fragilis sp. nov., A. gonzo sp. nov., A. muscosa Hickman, 1929, A. segmentata Simon, 1893, A. subnubilum sp. nov., A. thylacinus sp. nov. and A. tigrina sp. nov.. The species described in this manuscript exhibit high levels of sympatry.

Two new species of Western Australian Abantiades Herrich-Schffer (Lepidoptera: Hepialidae) and a description of the female of Abantiades paradoxa (Tindale).

Two new species, Abantiades concordia sp. nov. and Abantiades malleus sp. nov., are described from Australia. Both species were collected in the Eastern Goldfields subregion of the Coolgardie bioregion in Western Australia. Abantiades concordia sp. nov. is shown to be closely related to A. paradoxa (Tindale, 1932) by sequence similarity of the mtDNA (COI) gene. The female of A. paradoxa is also described here for the first time. Abantiades paradoxa and the new species A. concordia sp. nov. are morphologically similar with respect to the structure of their genitalia, sternite VIII, wing patterning and their antennae with bi-forked rami. Abantiades malleus sp. nov. is quite distinct by sequence similarity of the mtDNA (COI) gene, but related in a clade with A. marcidus Tindale,1932, A. albofasciatus (Swinhoe, 1892), and A. furva (Tindale,1932), the latter species once placed in the synonymised Bordaia Tindale, 1932. Discussion of similar species once grouped under the genus Bordaia and under the genus Trictena Meyrick, 1890 (both junior synonyms of Abantiades Herrich-Schffer, 1855) is also included.

Unparalleled mitochondrial heteroplasmy and Wolbachia co-infection in the non-model bee, Amphylaeus morosus.

Mitochondrial heteroplasmy is the occurrence of more than one type of mitochondrial DNA within a single individual. Although generally reported to occur in a small subset of individuals within a species, there are some instances of widespread heteroplasmy across entire populations. Amphylaeus morosus is an Australian native bee species in the diverse and cosmopolitan bee family Colletidae. This species has an extensive geographical range along the eastern Australian coast, from southern Queensland to western Victoria, covering approximately 2,000 km. Seventy individuals were collected from five localities across this geographical range and sequenced using Sanger sequencing for the mitochondrial cytochrome c oxidase subunit I (COI) gene. These data indicate that every individual had the same consistent heteroplasmic sites but no other nucleotide variation, suggesting two conserved and widespread heteroplasmic mitogenomes. Ion Torrent shotgun sequencing revealed that heteroplasmy occurred across multiple mitochondrial protein-coding genes and is unlikely explained by transposition of mitochondrial genes into the nuclear genome (NUMTs). DNA sequence data also demonstrated a consistent co-infection of Wolbachia across the A. morosus distribution with every individual infected with both bacterial strains. Our data are consistent with the presence of two mitogenomes within all individuals examined in this species and suggest a major divergence from standard patterns of mitochondrial inheritance. Because the host's mitogenome and the Wolbachia genome are genetically linked through maternal inheritance, we propose three possible hypotheses that could explain maintenance of the widespread and conserved co-occurring bacterial and mitochondrial genomes in this species.