Australian National Botanic Gardens

Abstract genalex is a user‐friendly cross‐platform package that runs within Microsoft Excel, enabling population genetic analyses of codominant, haploid and binary data. Allele frequency‐based analyses include heterozygosity, F statistics, Nei's genetic distance, population assignment, probabilities of identity and pairwise relatedness. Distance‐based calculations include amova , principal coordinates analysis (PCA), Mantel tests, multivariate and 2D spatial autocorrelation and twogener . More than 20 different graphs summarize data and aid exploration. Sequence and genotype data can be imported from automated sequencers, and exported to other software. Initially designed as tool for teaching, genalex 6 now offers features for researchers as well. Documentation and the program are available at http://www.anu.edu.au/BoZo/GenAlEx/

Birds are difficult to sex. Nestlings rarely show sex-linked morphology and we estimate that adult females appear identical to males in over 50% of the world's bird species. This problem can hinder both evolutionary studies and human-assisted breeding of birds. DNA-based sex identification provides a solution. We describe a test based on two conserved CHD (chromo-helicase-DNA-binding) genes that are located on the avian sex chromosomes of all birds, with the possible exception of the ratites (ostriches, etc.; Struthioniformes). The CHD-W gene is located on the W chromosome; therefore it is unique to females. The other gene, CHD-Z, is found on the Z chromosome and therefore occurs in both sexes (female, ZW; male, ZZ). The test employs PCR with a single set of primers. It amplifies homologous sections of both genes and incorporates introns whose lengths usually differ. When examined on a gel there is a single CHD-Z band in males but females have a second, distinctive CHD-W band.

Analyses of life‐history, ecological, and geographic trait differences among species, their causes, correlates, and likely consequences are increasingly important for understanding and conserving biodiversity in the face of rapid global change. Assembling multispecies trait data from diverse literature sources into a single comprehensive data set requires detailed consideration of methods to reliably compile data for particular species, and to derive single estimates from multiple sources based on different techniques and definitions. Here we describe PanTHERIA, a species‐level data set compiled for analysis of life history, ecology, and geography of all known extant and recently extinct mammals. PanTHERIA is derived from a database capable of holding multiple geo‐referenced values for variables within a species containing 100 740 lines of biological data for extant and recently extinct mammalian species, collected over a period of three years by 20 individuals. PanTHERIA also includes spatial databases of mammalian geographic ranges and global climatic and anthropogenic variables. Here we detail how the data fields are extracted and defined for PanTHERIA using a customized data input format (MammalForm); how data were collected from the literature, species names and sources tracked, error‐checking and validation procedures applied, and how data were consolidated into species‐level values for each variable. Tables of the consolidated species‐level values are made available for each of two recent species‐level taxonomic classifications of mammals, as well as associated taxonomic synonymy conversion and data‐input files. This study provides a useful guide to prospective researchers on how to structure and codify life‐history, ecological, geographic, and taxonomic data and methods to extract meaningful species‐level traits. It also provides comprehensive information on traits like size, diet, environmental conditions, and ecology to permit macroecological and macroevolutionary analyses of this important clade. The complete data sets corresponding to abstracts published in the Data Papers section of the journal are published electronically in Ecological Archives at 〈 http://esapubs.org/archive 〉. (The accession number for each Data Paper is given directly beneath the title.)

Aim To detect centres of vascular plant endemism at a continental scale by analysis of specimen‐based distributional data and to relate any pattern to environmental factors and history. Location Australia. Methods Presence of 8468 seed plant species‐level taxa throughout continental Australia and Tasmania was mapped on a 1° grid to visualize the pattern of species richness. This sample comprises half the known flora. Three indices of endemism were calculated but we preferred one that is unrelated to species richness, so that these two concepts could be distinguished in practice. Centres of endemism were detected by simple mapping and by spatial autocorrelation analysis (SAC). Linear regression was used to examine the relationship of the patterns of species richness and endemism to latitude, topography and climate. Results Both species richness and endemism vary greatly across the continent but in most cases the same centres were high in both richness and endemism. Twelve distinct centres were identified. The major centres of both diversity and endemism are south‐west western Australia, the Border Ranges between New South Wales and Queensland, the Wet Tropics near Cairns, Tasmania and the Iron‐McIlwraith Range of eastern Cape York Peninsula. The last centre appears to be more significant than recognized by past authors. Whether this is a true Australian centre of endemism, or is largely an outlier of the flora of Papua New Guinea, is explored. Another centre, in the Adelaide–Kangaroo Island region, has been overlooked altogether by previous authors. Regression analysis did not find a simple climatic explanation of the observed patterns. There was a suggestion that topographic variation within the 1° cells may be positively correlated with endemism, which is consistent with mountainous regions functioning as refugia. One clear result is that all the major centres of endemism are near‐coastal. A likely explanation is that Pleistocene expansions of the central desert have been a powerful limitation on the viability of refugia for narrowly endemic species. All the centres of endemism lie outside the estimated limits of the expanded arid zone at the last glacial maximum (18,000 yr BP ). In particular, the ‘Central Australian Mountain Ranges centre of plant diversity and endemism’ of Boden & Given (1995) is detected as a strong centre of species richness, but not at all as a centre of endemism. This is despite good sampling of this region. Main conclusions Endemism can be distinguished from species richness by using an appropriate index and mapping of such indices can detect centres of endemism. This study demonstrates the value of specimen based distributional data, such as is held in state herbaria and museums.

Dispersal is a fundamental process that influences the response of species to landscape change and habitat fragmentation. In an attempt to better understand dispersal in the Australian bush rat, Rattus fuscipes, we have combined a new multilocus autocorrelation method with hypervariable microsatellite genetic markers to investigate fine-scale (< or = 1 km) patterns of spatial distribution and spatial genetic structure. The study was conducted across eight trapping transects at four sites, with a total of 270 animals sampled. Spatial autocorrelation analysis of bush rat distribution revealed that, in general, animals occurred in groups or clusters of higher density (< or = 200 m across), with intervening gaps or lower density areas. Spatial genetic autocorrelation analysis, based on seven hypervariable microsatellite loci (He = 0.8) with a total of 80 alleles, revealed a consistent pattern of significant positive local genetic structure. This genetic pattern was consistent for all transects, and for adults and sub-adults, males and females. By testing for autocorrelation at multiple scales from 10 to 800 m we found that the extent of detectable positive spatial genetic structure exceeded 500 m. Further analyses detected significantly weaker spatial genetic structure in males compared with females, but no significant differences were detected between adults and sub adults. Results from Mantel tests and hierarchical AMOVA further support the conclusion that the distribution of bush rat genotypes is not random at the scale of our study. Instead, proximate bush rats are more genetically alike than more distant animals. We conclude that in bush rats, gene flow per generation is sufficiently restricted to generate the strong positive signal of local spatial genetic structure. Although our results are consistent with field data on animal movement, including the reported tendency for males to move further than females, we provide the first evidence for restricted gene flow in bush rats. Our study appears to be the first microsatellite-based study of fine-scale genetic variation in small mammals and the first to report consistent positive local genetic structure across sites, age-classes, and sexes. The combination of new forms of autocorrelation analyses, hypervariable genetic markers and fine-scale analysis (< 1 km) may thus offer new evolutionary insights that are overlooked by more traditional larger scaled (> 10 km) population genetic studies.

We investigated the transferability of 31 soybean (Glycine max) simple sequence repeat (SSR) loci to wild congeners and to other legume genera. Up to 65% of the soybean primer pairs amplified SSRs within Glycine, but frequently, the SSRs were short and interrupted compared with those of soybeans. Nevertheless, 85% of the loci were polymorphic within G. clandestina. Cross-species amplification outside of the genus was much lower (3%-13%), with polymorphism restricted to one primer pair, AG81. AG81 amplified loci in Glycine, Kennedia, and Vigna (Phaseoleae), Vicia (Vicieae), Trifolium (Trifolieae), and Lupinus (Genisteae) within the Papilionoideae, and in Albizia within the Mimosoideae. The primer conservation at AG81 may be explained by its apparent proximity to the seryl-tRNA synthetase gene. Interspecific differences in allele size at AG81 loci reflected repeat length variation within the SSR region and indels in the flanking region. Alleles of identical size with different underlying sequences (size homoplasy) were observed. Our findings and the emerging patterns in other plant studies suggest that in contrast to animals, successful cross-species amplification of SSRs in plants is largely restricted to congeners or closely related genera. Because mutations in both the SSR region and the flanking region contribute to variation in allele size among species, knowledge of DNA sequence is essential before SSR loci can be meaningfully used to address applied and evolutionary questions.

The plethora of studies devoted to the topics of male competition and female mate choice belie the fact that their interaction remains poorly understood. Indeed, on the question of whether competition should help or hinder the choice process, opinions scattered throughout the sexual selection literature seem unnecessarily polarised. We argue, in the light of recent theoretical and empirical advances, that the effect of competition on mate choice depends on whether it results in the choosy sex attaining high breeding value for total fitness, considering both direct and indirect fitness benefits. Specifically, trade-offs may occur between different fitness benefits if some are correlated with male competitive ability whilst others are not. Moreover, the costs and benefits of mating with competitive males may vary in time and/or space. These considerations highlight the importance of injecting a life-history perspective into sexual selection studies. Within this context, we turn to the sexual selection literature to try to offer insights into the circumstances when competition might be expected to have positive or negative implications for pre-copulatory female choice. In this regard, we elaborate on three stages where competition might impact upon the choice process: (i) during mate detection, (ii) mate evaluation, and (iii) in dictating actual mating outcomes. We conclude by offering researchers several potentially rewarding avenues for future research.

Theory suggests that species with particular traits are at greater risk of extinction than others. We assumed that a decline in abundance in forest fragments, compared to continuous forest, equated to an increase in extinction risk. We then tested the relationships between five traits of species and decline in abundance for 69 beetle species in an experimentally fragmented forest landscape at Mt. Wog Wog in southeastern Australia. The experiment was controlled and replicated. Monitoring ran for two years before forest fragmentation; in this paper, we examine data for five years postfragmentation. We tested five hypotheses: (1) Species that occur naturally at low abundance are more likely to decline as a result of fragmentation than are abundant species. (2) Isolated species are more likely to decline than species that are not isolated. (3) Large species are more likely to decline than small species. (4) Species in trophic groups at the top end of food chains are more likely to decline than species in trophic groups lower in the food chain. (5) Because traits are often shared by related species, populations of more closely related species will respond in the same way. We found that: (1) rare species were more likely to decline than abundant species; (2) isolated species were more likely to decline than species that were not isolated; (3) body size was not correlated with response to fragmentation; (4) among species that declined, predators declined most; and (5) taxonomically related species did not respond in the same way to fragmentation. Thus, our results confirm theories predicting that isolated, rare, or predaceous species will be lost first from fragmented landscapes.

Female lions (Panthera leo) showed persistent individual differences in the extent to which they participated in group-territorial conflict. When intergroup encounters were simulated by playback of aggressive vocalizations, some individuals consistently led the approach to the recorded intruder, whereas others lagged behind and avoided the risks of fighting. The lead females recognized that certain companions were laggards but failed to punish them, which suggests that cooperation is not maintained by reciprocity. Modification of the "odds" in these encounters revealed that some females joined the group response when they were most needed, whereas other lagged even farther behind. The complexity of these responses emphasizes the great diversity of individual behavior in this species and the inadequacy of current theory to explain cooperation in large groups.

Trait-based approaches have improved our understanding of plant evolution, community assembly and ecosystem functioning. A major challenge for the upcoming decades is to understand the functions and evolution of early life-history traits, across levels of organization and ecological strategies. Although a variety of seed traits are critical for dispersal, persistence, germination timing and seedling establishment, only seed mass has been considered systematically. Here we suggest broadening the range of morphological, physiological and biochemical seed traits to add new understanding on plant niches, population dynamics and community assembly. The diversity of seed traits and functions provides an important challenge that will require international collaboration in three areas of research. First, we present a conceptual framework for a seed ecological spectrum that builds upon current understanding of plant niches. We then lay the foundation for a seed-trait functional network, the establishment of which will underpin and facilitate trait-based inferences. Finally, we anticipate novel insights and challenges associated with incorporating diverse seed traits into predictive evolutionary ecology, community ecology and applied ecology. If the community invests in standardized seed-trait collection and the implementation of rigorous databases, major strides can be made at this exciting frontier of functional ecology.

Bacteriocins are an abundant class of antimicrobial molecules that appear to mediate population dynamics within species. The bacteriocins of Escherichia coli have served as a model for exploring the ecological role of these potent toxins. Studies suggest that colicins provide a competitive edge in nutrient-poor environments and that there might be a trade-off between the costs and benefits of colicin production.

Many unusual features of scale insects (Hemiptera: Coccoidea) can be explained as historical legacy. Developmental specializations in ancestral coccoids resulted in a neotenous adult female and a drastic metamorphosis of the male. Subsequent evolution led to numerous, often convergently derived, adaptations to parasitic life on higher plants. The sedentary lifestyle of female scale insects has favored the evolution of appendage reduction or loss, gross changes in body shape, and protective wax secretions, tests, and other scale covers. Morphological peculiarities of adult males relate to flight or to mating with concealed females. Scale insects have diverse egg-protecting methods, a range of chromosome behaviors (including several methods of sex determination), marked sexual dimorphism [even sometimes in first-instar nymphs (crawlers)], and more rarely sexual dichronism. Crawlers have evolved as the main agents of dispersal. The biotic interactions of scale insects include diverse endosymbioses with microorganisms, sometimes morphological and behavioral adaptations for obligate association with ants, and often highly specific host-plant associations that may lead to demic adaptation or the evolution of complex galls.

Theory and empirical evidence have long suggested that some species are extremely vulnerable to extinction because they have combinations of extinction promoting traits. However, ecologists have not considered whether the form of the relationship between traits is additive (not synergistic) or nonadditive (synergistic). We looked at how traits and their interactions were related to the difference in species' population growth rates between experimentally fragmented forest and continuous forest. Two traits acted synergistically; natural abundance and degree of specialization interacted so that beetle species that were rare and specialized had a greater reduction in their growth rates in fragments, compared to continuous forest, than the sum of the reductions in growth rates attributable to these traits. In other words, species that were both rare and specialized were especially vulnerable to extinction. From a conservation perspective, an implication of our findings is that making predictions about extinction risk from a single trait, like abundance or population variability, may be risky because traits may act synergistically rendering species more, or less, vulnerable than predicted by that single trait. There is currently a great deal of interest in which traits predict the sensitivity of species to a given threat, but if we are going to look at risks in this way, then we also need to consider how traits interact, because this can alter the vulnerability of species.

Top predators in terrestrial ecosystems may limit populations of smaller predators that could otherwise become over abundant and cause declines and extinctions of some prey. It is therefore possible that top predators indirectly protect many species of prey from excessive predation. This effect has been demonstrated in some small-scale studies, but it is not known how general or important it is in maintaining prey biodiversity. During the last 150 years, Australia has suffered the world's highest rate of mammal decline and extinction, and most evidence points to introduced mid-sized predators (the red fox and the feral cat) as the cause. Here, we test the idea that the decline of Australia's largest native predator, the dingo, played a role in these extinctions. Dingoes were persecuted from the beginning of European settlement in Australia and have been eliminated or made rare over large parts of the continent. We show a strong positive relationship between the survival of marsupials and the geographical overlap with high-density dingo populations. Our results suggest that the rarity of dingoes was a critical factor which allowed smaller predators to overwhelm marsupial prey, triggering extinction over much of the continent. This is evidence of a crucial role of top predators in maintaining prey biodiversity at large scales in terrestrial ecosystems and suggests that many remaining Australian mammals would benefit from the positive management of dingoes.

Chloroplast microsatellites, or simple sequence repeats (cpSSRs), are typically mononucleotide tandem repeats. When located in the noncoding regions of the chloroplast genome (cpDNA), they commonly show intraspecific variation in repeat number. Despite the growing number of studies applying cpSSRs, studies of economically important plants and their relatives remain over-represented. Thus, the potential of cpSSRs to offer unique insights into ecological and evolutionary processes in wild plant species has yet to be fully realized. This review provides an overview of the technical resources available to aid cpSSR discovery including a list of cpSSR primer sets available and cpDNA sequencing resources. Our updated analysis of 99 whole chloroplast genomes downloaded from GenBank confirms that potentially variable cpSSRs are abundant in the noncoding cpDNA of plants. Overall variation in the frequency of cpSSRs was extreme, ranging from one to 700 per genome (median = 93), while in 81 vascular plants, between 35 and 160 cpSSRs were detected per genome (median = 86). We offer five recommendations to aid wider development and application of cpSSRs: (i) When genus-specific cpSSR primers are available, cross-species amplification can often be fruitful. (ii) While potentially useful, universal cpSSR primers at best provide access to only a small number of variable markers. (iii) De novo sequencing of noncoding cpDNA is the most effective and efficient way to develop cpSSR markers in wild species. (iv) DNA sequencing of cpSSR alleles is essential, given the complex nature of the genetic variation associated with hypervariable cpDNA regions. (v) The reliability of cpSSR length based genetic assays need to be validated in all studies.

The availability of soybean mutants with altered symbiotic properties allowed an investigation of the shoot or root control of the relevant phenotype. By means of grafts between these mutants and wild-type plants (cultivar Bragg and Williams), we demonstrated that supernodulation as well as hypernodulation (nitrate tolerance in nodulation and lack of autoregulation) is shoot controlled in two mutants (nts382 and nts1116) belonging most likely to two separate complementation groups. The supernodulation phenotype was expressed on roots of the parent cultivar Bragg as well as the roots of cultivar Williams. Likewise it was shown that non-nodulation (resistance to Bradyrhizobium) is root controlled in mutant nod49. The shoot control of nodule initiation is epistatically suppressed by the non-nodulation, root-expressed mutation. These findings suggest that different plant organs can influence the expression of the nodulation phenotype.

Recent attempts to explain the susceptibility of vertebrates to declines worldwide have largely focused on intrinsic factors such as body size, reproductive potential, ecological specialization, geographical range and phylogenetic longevity. Here, we use a database of 145 Australian marsupial species to test the effects of both intrinsic and extrinsic factors in a multivariate comparative approach. We model five intrinsic (body size, habitat specialization, diet, reproductive rate and range size) and four extrinsic (climate and range overlap with introduced foxes, sheep and rabbits) factors. We use quantitative measures of geographical range contraction as indices of decline. We also develop a new modelling approach of phylogenetically independent contrasts combined with imputation of missing values to deal simultaneously with phylogenetic structuring and missing data. One extrinsic variable-geographical range overlap with sheep-was the only consistent predictor of declines. Habitat specialization was independently but less consistently associated with declines. This suggests that extrinsic factors largely determine interspecific variation in extinction risk among Australian marsupials, and that the intrinsic factors that are consistently associated with extinction risk in other vertebrates are less important in this group. We conclude that recent anthropogenic changes have been profound enough to affect species on a continent-wide scale, regardless of their intrinsic biology.

It is well recognized that Escherichia coli consists of a number of distinct phylo-groups and that strains of the different phylo-groups vary in their ecological niches, life-history characteristics and propensity to cause disease. Consequently, much can be learnt by assigning a strain of E. coli to one of the recognized phylo-groups. A triplex PCR-based method that enables strains of E. coli to be assigned to a phylo-group using a dichotomous key approach based on the presence or absence of two genes (chuA and yjaA) and an anonymous DNA fragment (TSPE4.C2) has been developed. However, the accuracy with which this method assigns strains to their correct phylo-group has not been adequately evaluated. Consequently, 662 strains of E. coli were characterized using a multi-locus sequence typing approach. Unsupervised population assignment algorithms were used to assign strains to phylo-groups based on the multi-locus sequence typing data. The analyses revealed that 85-90% of E. coli strains can be assigned to a phylo-group and that 80-85% of the phylo-group memberships assigned using the Clermont method are correct. However, the accuracy with which strains are assigned to the correct phylo-group depends on their Clermont genotype. For example, strains yielding a Clermont genotype consistent with phylo-groups B1 and B2 are assigned correctly 95% of the time. Strains failing to yield any PCR products using the Clermont method are seldom members of phylo-group A and strains with such a genotype should not be assigned to a phylo-group.

Summary Ecologists have long recognized that plants occurring in areas of low rainfall or soil nutrients tend to have smaller leaves than those in more favourable regions. Working with a large data set (690 species at 47 sites spread widely through south‐east Australia) for which this reduction has been described previously, we investigated the morphology of leaf size reduction, asking whether any patterns observed were consistent across evolutionary lineages or between environmental gradients. Leaf length, width and surface areas were measured; leaf traits such as pubescence or lobing were also scored qualitatively. There was no correlation between soil phosphorus and rainfall across sites. Further, there was no evidence that pubescence, lobing or other traits assessed served as alternatives to reduction of leaf size at the low ends of either environmental gradient. Leaf size reduction occurred through many combinations of change in leaf width and length, even within lineages. Thus consistent patterns in the method of leaf size reduction were not found, although broad similarities between rainfall and soil P gradients were apparent.

Recent reviews of specific topics, such as the relationship between male attractiveness to females and fluctuating asymmetry or attractiveness and the expression of secondary sexual characters, suggest that publication bias might be a problem in ecology and evolution. In these cases, there is a significant negative correlation between the sample size of published studies and the magnitude or strength of the research findings (formally the 'effect size'). If all studies that are conducted are equally likely to be published, irrespective of their findings, there should not be a directional relationship between effect size and sample size; only a decrease in the variance in effect size as sample size increases due to a reduction in sampling error. One interpretation of these reports of negative correlations is that studies with small sample sizes and weaker findings (smaller effect sizes) are less likely to be published. If the biological literature is systematically biased this could undermine the attempts of reviewers to summarise actual biology relationships by inflating estimates of average effect sizes. But how common is this problem? And does it really affect the general conclusions of'literature reviews? Here, we examine data sets of effect sizes extracted from 40 peer-reviewed, published meta-analyses. We estimate how many studies are missing using the newly developed 'trim and fill' method. This method uses asymmetry in plots of effect size against sample size ('funnel plots') to detect missing' studies. For random-effect models of meta-analysis 38% (15/40) of data sets had a significant number of 'missing' studies. After correcting for potential publication bias, 21 % (8/38) of weighted mean effects were no longer significantly greater than zero, and 15% (5/34) were no longer statistically robust when we used random-effects models in a weighted meta-analysis. The mean correlation between sample size and the magnitude of standardised effect size was also significantly negative (r(s) = -0.20, P < 0.0001). Individual correlations were significantly negative (P<0.10) in 35% (14/40) of cases. Publication bias may therefore affect the main conclusions of at least 15-21% of meta-analyses. We suggest that future literature reviews assess the robustness of their main conclusions by correcting for potential publication bias using the 'trim and fill' method.