Mitochondrial DNA phylogeography of the Mesoamerican spiny-tailed lizards (Ctenosaura quinquecarinata complex): historical biogeography, species status and conservation

Through the examination of past and present distributions of plants and animals, historical biogeographers have provided many insights on the dynamics of the massive organismal exchange between North and South America. However, relatively few phylogeographic studies have been attempted in the land bridge of Mesoamerica despite its importance to better understand the evolutionary forces influencing this biodiversity 'hotspot'. Here we use mitochondrial DNA sequence data from fresh samples and formalin-fixed museum specimens to investigate the genetic and biogeographic diversity of the threatened Mesoamerican spiny-tailed lizards of the Ctenosaura quinquecarinata complex. Species boundaries and their phylogeographic patterns are examined to better understand their disjunct distribution. Three monophyletic, allopatric lineages are established using mtDNA phylogenetic and nested clade analyses in (i) northern: México, (ii) central: Guatemala, El Salvador and Honduras, and (iii) southern: Nicaragua and Costa Rica. The average sequence divergence observed between lineages varied between 2.0% and 3.7% indicating that they do not represent a very recent split and the patterns of divergence support the recently established nomenclature of C. quinquecarinata, Ctenosaura flavidorsalis and Ctenosaura oaxacana. Considering the geological history of Mesoamerica and the observed phylogeographic patterns of these lizards, major evolutionary episodes of their radiation in Mesoamerica are postulated and are indicative of the regions' geological complexity. The implications of these findings for the historical biogeography, taxonomy and conservation of these lizards are discussed.     

Analysing botanical collecting effort in Amazonia and correcting for it in species range estimation

Aim The aim of this study is to analyse the distribution pattern of the botanical collecting effort in Amazonia so that it can be accounted for when interpreting phytogeographical patterns such as inferred species ranges. We also develop a mechanistic and transparent method for taking into account the bias in collecting effort when estimating likelihoods of species occurrences. Location Amazonia, Neotropics. Methods We utilized electronic data sets of georeferenced herbarium collections (1,063,530 in total). We plotted collecting localities (68,246 in total) on maps overlaid with 1° and 0.5° square grids, and analysed collecting effort using a geographical information system (GIS). We also drew a map of Thiessen polygons, using collecting localities as polygon centres, to visualize collecting density in a scale‐independent way. We then created a ‘collecting activity landscape’ in which well‐collected areas appear as peaks and poorly studied areas as valleys. We demonstrate how this surface can be utilized when estimating species distributions. Results The data available to us confirm that botanical collecting activity is still severely biased in Amazonia. The uncollected area represents 43% of the total area of Amazonia, while another 28% is poorly collected and only 2% can be considered relatively well collected. The Thiessen polygon network represents an improvement in the presentation of collecting intensity compared with square grids. Main conclusions The maps of botanical collecting effort in the Neotropics should be used for visually correcting phytogeographical interpretations. With the help of GIS applications the observed spatial bias in collecting effort can be utilized in estimating the likelihood of occurrence of species in a repeatable manner. These estimates, in turn, can be used for various purposes in basic and applied science as well as in decision‐making. The biased collecting effort should, in the long run, be corrected by further field work in unexplored areas, which can be identified with the maps presented here.     

Sensitivity of predictive species distribution models to change in grain size

Predictive species distribution modelling (SDM) has become an essential tool in biodiversity conservation and management. The choice of grain size (resolution) of environmental layers used in modelling is one important factor that may affect predictions. We applied 10 distinct modelling techniques to presence-only data for 50 species in five different regions, to test whether: (1) a 10-fold coarsening of resolution affects predictive performance of SDMs, and (2) any observed effects are dependent on the type of region, modelling technique, or species considered. Results show that a 10 times change in grain size does not severely affect predictions from species distribution models. The overall trend is towards degradation of model performance, but improvement can also be observed. Changing grain size does not equally affect models across regions, techniques, and species types. The strongest effect is on regions and species types, with tree species in the data sets (regions) with highest locational accuracy being most affected. Changing grain size had little influence on the ranking of techniques: boosted regression trees remain best at both resolutions. The number of occurrences used for model training had an important effect, with larger sample sizes resulting in better models, which tended to be more sensitive to grain. Effect of grain change was only noticeable for models reaching sufficient performance and/or with initial data that have an intrinsic error smaller than the coarser grain size.     

Whole-drawer imaging for digital management and curation of a large entomological collection

Whole-drawer imaging is shown to be an effective tool for rapid digitisation of large insect collections. On-line, Whole-drawer images facilitate more effective collection management, virtual curation, and public engagement. The Whole-drawer imaging experience at the Australian National Insect Collection is discussed, with an explanation of workflow and examples of benefits.     

VARIABILITY IN TEMPERATURE AND HISTORICAL PATTERNS IN REPRODUCTION IN THE FUCUS DISTICHUS COMPLEX (HETEROKONTOPHYTA; PHAEOPHYCEAE): IMPLICATIONS FOR SPECIATION AND THE COLLECTION OF HERBARIUM SPECIMENS1

Natural and anthropogenic‐induced fluctuations in temperature can have profound effects on an organism's reproduction. The potential of herbarium collections for detecting changes in the timing of reproduction associated with interannual fluctuations in temperature from 1905 to 2003 was assessed in several taxa in the Fucus distichus (L.) emend. Powell complex from northern New England. Data were also used to evaluate whether air and/or sea temperature and North Atlantic Oscillation indices can cause overlap in the reproductive seasonalities of these taxa, a potential mechanism that prevents speciation. For all taxa, there was much variability in reproductive seasonalities among years. Discerning potential correlations with air or sea temperature was difficult because of a suspected bias to collect reproductive specimens at times when most of the population was nonreproductive. Nevertheless, there was a trend for seawater temperature to explain aseasonal reproduction for some taxa. Moreover, the existing archive (1895–2003) shows that the reproductive seasons of these taxa often overlap, and we suggest that interannual variability in temperature may often inhibit speciation. Suggestions also include archive of both reproductive and vegetative materials as well as coordinated sampling in critical biogeographic regions. No one investigator or institution can accomplish this, but phycological societies could organize and sustain such efforts.     

Phylogenomics using formalin‐fixed and 100+ year‐old intractable natural history specimens

Museum specimens provide a wealth of information to biologists, but obtaining genetic data from formalin‐fixed and fluid‐preserved specimens remains challenging. While DNA sequences have been recovered from such specimens, most approaches are time‐consuming and produce low data quality and quantity. Here, we use a modified DNA extraction protocol combined with high‐throughput sequencing to recover DNA from formalin‐fixed and fluid‐preserved snakes that were collected over a century ago and for which little or no modern genetic materials exist in public collections. We successfully extracted DNA and sequenced ultraconserved elements ( = 2318 loci) from 10 fluid‐preserved snakes and included them in a phylogeny with modern samples. This phylogeny demonstrates the general use of such specimens in phylogenomic studies and provides evidence for the placement of enigmatic snakes, such as the rare and never‐before sequenced Indian Xylophis stenorhynchus. Our study emphasizes the relevance of museum collections in modern research and simultaneously provides a protocol that may prove useful for specimens that have been previously intractable for DNA sequencing.     

Museum genomics: low-cost and high-accuracy genetic data from historical specimens

Natural history collections are unparalleled repositories of geographical and temporal variation in faunal conditions. Molecular studies offer an opportunity to uncover much of this variation; however, genetic studies of historical museum specimens typically rely on extracting highly degraded and chemically modified DNA samples from skins, skulls or other dried samples. Despite this limitation, obtaining short fragments of DNA sequences using traditional PCR amplification of DNA has been the primary method for genetic study of historical specimens. Few laboratories have succeeded in obtaining genome-scale sequences from historical specimens and then only with considerable effort and cost. Here, we describe a low-cost approach using high-throughput next-generation sequencing to obtain reliable genome-scale sequence data from a traditionally preserved mammal skin and skull using a simple extraction protocol. We show that single-nucleotide polymorphisms (SNPs) from the genome sequences obtained independently from the skin and from the skull are highly repeatable compared to a reference genome.     

Unlocking the vault: next‐generation museum population genomics

Natural history museum collections provide unique resources for understanding how species respond to environmental change, including the abrupt, anthropogenic climate change of the past century. Ideally, researchers would conduct genome‐scale screening of museum specimens to explore the evolutionary consequences of environmental changes, but to date such analyses have been severely limited by the numerous challenges of working with the highly degraded DNA typical of historic samples. Here, we circumvent these challenges by using custom, multiplexed, exon capture to enrich and sequence ~11 000 exons (~4 Mb) from early 20th‐century museum skins. We used this approach to test for changes in genomic diversity accompanying a climate‐related range retraction in the alpine chipmunks (Tamias alpinus) in the high Sierra Nevada area of California, USA. We developed robust bioinformatic pipelines that rigorously detect and filter out base misincorporations in DNA derived from skins, most of which likely resulted from postmortem damage. Furthermore, to accommodate genotyping uncertainties associated with low‐medium coverage data, we applied a recently developed probabilistic method to call single‐nucleotide polymorphisms and estimate allele frequencies and the joint site frequency spectrum. Our results show increased genetic subdivision following range retraction, but no change in overall genetic diversity at either nonsynonymous or synonymous sites. This case study showcases the advantages of integrating emerging genomic and statistical tools in museum collection‐based population genomic applications. Such technical advances greatly enhance the value of museum collections, even where a pre‐existing reference is lacking and points to a broad range of potential applications in evolutionary and conservation biology.     

Long-Term Storage of Infective Microsporidian Spores in Liquid Nitrogen

Thirty-one species of microsporidia, isolated from insects and stored in liquid nitrogen for up to 25 yr, were infectious when removed from liquid nitrogen. The natural hosts of all of these microsporidia were terrestrial insects, representing six different insect orders: Coleoptera, Diptera, Hemiptera, Hymenoptera, Lepidoptera, and Orthoptera. All microsporidia from terrestrial insects that were tested survived storage in liquid nitrogen, while Nosema algerae , a microsporidium from aquatic mosquito hosts did not survive freezing in liquid nitrogen. A Nosema species from the alfalfa weevil, Hypera postica , lost some infectivity in a water storage medium after 25 yr in liquid nitrogen. Liquid nitrogen storage of microsporidian spores in 50% and 100% glycerol media reduced loss of infectivity and is recommended for extended storage of microsporidia from terrestrial insect hosts.