Discrimination of Hover Fly Species and Sexes by Wing Interference Signals.

Remote automated surveillance of insect abundance and diversity is poised to revolutionize insect decline studies. The study reveals spectral analysis of thin-film wing interference signals (WISs) can discriminate free-flying insects beyond what can be accomplished by machine vision. Detectable by photonic sensors, WISs are robust indicators enabling species and sex identification. The first quantitative survey of insect wing thickness and modulation through shortwave-infrared hyperspectral imaging of 600 wings from 30 hover fly species is presented. Fringy spectral reflectance of WIS can be explained by four optical parameters, including membrane thickness. Using a Naïve Bayes Classifier with five parameters that can be retrieved remotely, 91% is achieved accuracy in identification of species and sexes. WIS-based surveillance is therefore a potent tool for remote insect identification and surveillance.

A biophotonic platform for quantitative analysis in the spatial, spectral, polarimetric, and goniometric domains.

Advanced instrumentation and versatile setups are needed for understanding light interaction with biological targets. Such instruments include (1) microscopes and 3D scanners for detailed spatial analysis, (2) spectral instruments for deducing molecular composition, (3) polarimeters for assessing structural properties, and (4) goniometers probing the scattering phase function of, e.g., tissue slabs. While a large selection of commercial biophotonic instruments and laboratory equipment are available, they are often bulky and expensive. Therefore, they remain inaccessible for secondary education, hobbyists, and research groups in low-income countries. This lack of equipment impedes hands-on proficiency with basic biophotonic principles and the ability to solve local problems with applied physics. We have designed, prototyped, and evaluated the low-cost Biophotonics, Imaging, Optical, Spectral, Polarimetric, Angular, and Compact Equipment (BIOSPACE) for high-quality quantitative analysis. BIOSPACE uses multiplexed light-emitting diodes with emission wavelengths from ultraviolet to near-infrared, captured by a synchronized camera. The angles of the light source, the target, and the polarization filters are automated by low-cost mechanics and a microcomputer. This enables multi-dimensional scatter analysis of centimeter-sized biological targets. We present the construction, calibration, and evaluation of BIOSPACE. The diverse functions of BIOSPACE include small animal spectral imaging, measuring the nanometer thickness of a bark-beetle wing, acquiring the scattering phase function of a blood smear and estimating the anisotropic scattering and the extinction coefficients, and contrasting muscle fibers using polarization. We provide blueprints, component list, and software for replication by enthusiasts and educators to simplify the hands-on investigation of fundamental optical properties in biological samples.

Potential for identification of wild night-flying moths by remote infrared microscopy.

There are hundreds of thousands of moth species with crucial ecological roles that are often obscured by their nocturnal lifestyles. The pigmentation and appearance of moths are dominated by cryptic diffuse shades of brown. In this study, 82 specimens representing 26 moth species were analysed using infrared polarimetric hyperspectral imaging in the range of 0.95-2.5 µm. Contrary to previous studies, we demonstrate that since infrared light does not resolve the surface roughness, wings appear glossy and specular at longer wavelengths. Such properties provide unique reflectance spectra between species. The reflectance of the majority of our species could be explained by comprehensive models, and a complete parametrization of the spectral, polarimetric and angular optical properties was reduced to just 11 parameters with physical units. These parameters are complementary and, compared with the within-species variation, were significantly distinct between species. Counterintuitively to the aperture-limited resolution criterion, we could deduce microscopic features along the surface from their infrared properties. These features were confirmed by electron microscopy. Finally, we show how our findings could greatly enhance opportunities for remote identification of free-flying moth species, and we hypothesize that such flat specular wing targets could be expected to be sensed over considerable distances.

A simple method for data partitioning based on relative evolutionary rates.

BACKGROUND: Multiple studies have demonstrated that partitioning of molecular datasets is important in model-based phylogenetic analyses. Commonly, partitioning is done a priori based on some known properties of sequence evolution, e.g. differences in rate of evolution among codon positions of a protein-coding gene. Here we propose a new method for data partitioning based on relative evolutionary rates of the sites in the alignment of the dataset being analysed. The rates are inferred using the previously published Tree Independent Generation of Evolutionary Rates (TIGER), and the partitioning is conducted using our novel python script RatePartitions. We conducted simulations to assess the performance of our new method, and we applied it to eight published multi-locus phylogenetic datasets, representing different taxonomic ranks within the insect order Lepidoptera (butterflies and moths) and one phylogenomic dataset, which included ultra-conserved elements as well as introns. METHODS: We used TIGER-rates to generate relative evolutionary rates for all sites in the alignments. Then, using RatePartitions, we partitioned the data into partitions based on their relative evolutionary rate. RatePartitions applies a simple formula that ensures a distribution of sites into partitions following the distribution of rates of the characters from the full dataset. This ensures that the invariable sites are placed in a partition with slowly evolving sites, avoiding the pitfalls of previously used methods, such as k-means. Different partitioning strategies were evaluated using BIC scores as calculated by PartitionFinder. RESULTS: Simulations did not highlight any misbehaviour of our partitioning approach, even under difficult parameter conditions or missing data. In all eight phylogenetic datasets, partitioning using TIGER-rates and RatePartitions was significantly better as measured by the BIC scores than other partitioning strategies, such as the commonly used partitioning by gene and codon position. We compared the resulting topologies and node support for these eight datasets as well as for the phylogenomic dataset. DISCUSSION: We developed a new method of partitioning phylogenetic datasets without using any prior knowledge (e.g. DNA sequence evolution). This method is entirely based on the properties of the data being analysed and can be applied to DNA sequences (protein-coding, introns, ultra-conserved elements), protein sequences, as well as morphological characters. A likely explanation for why our method performs better than other tested partitioning strategies is that it accounts for the heterogeneity in the data to a much greater extent than when data are simply subdivided based on prior knowledge.

Community structure of insect herbivores is driven by conservatism, escalation and divergence of defensive traits in Ficus.

Escalation (macroevolutionary increase) or divergence (disparity between relatives) in trait values are two frequent outcomes of the plant-herbivore arms race. We studied the defences and caterpillars associated with 21 sympatric New Guinean figs. Herbivore generalists were concentrated on hosts with low protease and oxidative activity. The distribution of specialists correlated with phylogeny, protease and trichomes. Additionally, highly specialised Asota moths used alkaloid rich plants. The evolution of proteases was conserved, alkaloid diversity has escalated across the studied species, oxidative activity has escalated within one clade, and trichomes have diverged across the phylogeny. Herbivore specificity correlated with their response to host defences: escalating traits largely affected generalists and divergent traits specialists; but the effect of escalating traits on extreme specialists was positive. In turn, the evolution of defences in Ficus can be driven towards both escalation and divergence in individual traits, in combination providing protection against a broad spectrum of herbivores.

PCR primers for 30 novel gene regions in the nuclear genomes of Lepidoptera.

We report primer pairs for 30 new gene regions in the nuclear genomes of Lepidoptera that can be amplified using a standard PCR protocol. The new primers were tested across diverse Lepidoptera, including nonditrysians and a wide selection of ditrysians. These new gene regions give a total of 11,043 bp of DNA sequence data and they show similar variability to traditionally used nuclear gene regions in studies of Lepidoptera. We feel that a PCR-based approach still has its place in molecular systematic studies of Lepidoptera, particularly at the intrafamilial level, and our new set of primers now provides a route to generating phylogenomic datasets using traditional methods.

Integrative Taxonomy Recognizes Evolutionary Units Despite Widespread Mitonuclear Discordance: Evidence from a Rotifer Cryptic Species Complex.

Mitonuclear discordance across taxa is increasingly recognized as posing a major challenge to species delimitation based on DNA sequence data. Integrative taxonomy has been proposed as a promising framework to help address this problem. However, we still lack compelling empirical evidence scrutinizing the efficacy of integrative taxonomy in relation to, for instance, complex introgression scenarios involving many species. Here, we report remarkably widespread mitonuclear discordance between about 15 mitochondrial and 4 nuclear Brachionus calyciflorus groups identified using different species delimitation approaches. Using coalescent-, Bayesian admixture-, and allele sharing-based methods with DNA sequence or microsatellite data, we provide strong evidence in support of hybridization as a driver of the observed discordance. We then describe our combined molecular, morphological, and ecological approaches to resolving phylogenetic conflict and inferring species boundaries. Species delimitations based on the ITS1 and 28S nuclear DNA markers proved a more reliable predictor of morphological variation than delimitations using the mitochondrial COI gene. A short-term competition experiment further revealed systematic differences in the competitive ability between two of the nuclear-delimited species under six different growth conditions, independent of COI delimitations; hybrids were also observed. In light of these findings, we discuss the failure of the COI marker to estimate morphological stasis and morphological plasticity in the B. calyciflorus complex. By using B. calyciflorus as a representative case, we demonstrate the potential of integrative taxonomy to guide species delimitation in the presence of mitonuclear phylogenetic conflicts.

The butterfly plant arms-race escalated by gene and genome duplications.

Coevolutionary interactions are thought to have spurred the evolution of key innovations and driven the diversification of much of life on Earth. However, the genetic and evolutionary basis of the innovations that facilitate such interactions remains poorly understood. We examined the coevolutionary interactions between plants (Brassicales) and butterflies (Pieridae), and uncovered evidence for an escalating evolutionary arms-race. Although gradual changes in trait complexity appear to have been facilitated by allelic turnover, key innovations are associated with gene and genome duplications. Furthermore, we show that the origins of both chemical defenses and of molecular counter adaptations were associated with shifts in diversification rates during the arms-race. These findings provide an important connection between the origins of biodiversity, coevolution, and the role of gene and genome duplications as a substrate for novel traits.

Notable plesiomorphies and notable specializations: head structure of the primitive "tongue moth" Acanthopteroctetes unifascia (Lepidoptera: Acanthopteroctetidae).

The Acanthopteroctetidae are one of the first-originated family-group lineages within "tongue moths" (Lepidoptera-Glossata). The purpose of this study is to provide a comprehensive account (based on whole mount preparations, serial sections, and Scanning electron microscopy) of the cephalic structure of an adult exemplar of the family, to supplement the sparse available information. Notable plesiomorphies include the retention of frontal retractors of the narrow labrum, a high supraocular index linked to strong development of cranio-mandibular ad- and abductors, and perhaps the unusually short but still coilable (just ca. 1.5 turns) galeal "tongue." Notable specializations (probably mostly family autapomorphies) include a complement of large sensilla placodea on the male antennae, an apical attachment of the long dorsal tentorial arm to the cranium, an extreme reduction of the single-segmented labial palps, a particularly strong subgenal bridge and a surface structure of near-parallel ridges on the ommatidial corneae. The presence of sizable saccular mandibular (type 1) glands opening into the adductor apodeme is unexpected, no counterparts being known from neighboring taxa. The same is true for ventral salivarium dilator muscles originating on the prelabium; and tentatively suggested to be homologues of the extrinsic palp flexors (the insertion shift being related to loss of original function due to palp reduction), rather than to the ventral salivarium muscles of more basal insects. A complete "deutocerebral loop"' may or may not be developed, as is true for a mutual appression of the optic lobe and circumoesophageal connective/suboesophageal ganglion, enclosing the anterior tentorial arm between them; a suboesophageal innervation of the retrocerebral complex was not observed. No characters bearing on the monophyly of the Coelolepida were identified. The scapo-pedicellar articulation with a scapal process and a smooth intercalary sclerite is reminiscent of conditions in Neopseustidae, but remains debatable as a synapomorphy of the two families.

A new genus of metalmark moths (Lepidoptera, Choreutidae) with Afrotropical and Australasian distribution.

Niveas Rota, new genus, and its two new species, N. agassizi Rota, new species, and N. kone Rota, new species, are described and illustrated. Niveas is assigned to the subfamily Choreutinae based on morphological and molecular data. Niveas agassizi is currently known only from Kenya and only from female specimens. Niveas kone has been found on the Solomon Islands and in Papua New Guinea (PNG). In PNG, larvae of this species have been reared from several species of Ficus (Moraceae). The two species are superficially quite dissimilar from each other. However, they share features in wing pattern and venation, as well as female genitalia, and the molecular data strongly support the monophyly of Niveas.

Tortricid moths reared from the invasive weed Mexican palo verde, Parkinsonia aculeata, with comments on their host specificity, biology, geographic distribution, and systematics.

As part of efforts to identify native herbivores of Mexican palo verde, Parkinsonia aculeata L. (Leguminosae: Caesalpinioideae), as potential biological control agents against this invasive weed in Australia, ten species of Tortricidae (Lepidoptera) were reared from Guatemala, Mexico, Nicaragua, and Venezuela: Amorbia concavana (Zeller), Platynota rostrana (Walker), Platynota helianthes (Meyrick), Platynota stultana Walsingham (all Tortricinae: Sparganothini), Rudenia leguminana (Busck), Cochylis sp. (both Tortricinae: Cochylini), Ofatulena duodecemstriata (Walsingham), O. luminosa Heinrich, Ofatulena sp. (all Olethreutinae: Grapholitini), and Crocidosema lantana Busck (Olethreutinae: Eucosmini). Significant geographic range extensions are provided for O. duodecemstriata and R. leguminana. These are the first documented records of P. aculeata as a host plant for all but O. luminosa. The four species of Sparganothini are polyphagous; in contrast, the two Cochylini and three Grapholitini likely are specialists on Leguminosae. Ofatulena luminosa is possibly host specific on P. aculeata. Host trials with Rudenia leguminana also provide some evidence of specificity, in contrast to historical rearing records. To examine the possibility that R. leguminana is a complex of species, two data sets of molecular markers were examined: (1) a combined data set of two mitochondrial markers (a 781-basepair region of cytochrome c oxidase I (COI) and a 685-basepair region of cytochrome c oxidase II) and one nuclear marker (a 531-basepair region of the 28S domain 2); and (2) the 650-basepair "barcode" region of COI. Analyses of both data sets strongly suggest that individuals examined in this study belong to more than one species.

Predator mimicry: metalmark moths mimic their jumping spider predators.

Cases of mimicry provide many of the nature's most convincing examples of natural selection. Here we report evidence for a case of predator mimicry in which metalmark moths in the genus Brenthia mimic jumping spiders, one of their predators. In controlled trials, Brenthia had higher survival rates than other similarly sized moths in the presence of jumping spiders and jumping spiders responded to Brenthia with territorial displays, indicating that Brenthia were sometimes mistaken for jumping spiders, and not recognized as prey. Our experimental results and a review of wing patterns of other insects indicate that jumping spider mimicry is more widespread than heretofore appreciated, and that jumping spiders are probably an important selective pressure shaping the evolution of diurnal insects that perch on vegetation.