In vivo visualization of butterfly scale cell morphogenesis in Vanessa cardui.

During metamorphosis, the wings of a butterfly sprout hundreds of thousands of scales with intricate microstructures and nano-structures that determine the wings' optical appearance, wetting characteristics, thermodynamic properties, and aerodynamic behavior. Although the functional characteristics of scales are well known and prove desirable in various applications, the dynamic processes and temporal coordination required to sculpt the scales' many structural features remain poorly understood. Current knowledge of scale growth is primarily gained from ex vivo studies of fixed scale cells at discrete time points; to fully understand scale formation, it is critical to characterize the time-dependent morphological changes throughout their development. Here, we report the continuous, in vivo, label-free imaging of growing scale cells of Vanessa cardui using speckle-correlation reflection phase microscopy. By capturing time-resolved volumetric tissue data together with nanoscale surface height information, we establish a morphological timeline of wing scale formation and gain quantitative insights into the underlying processes involved in scale cell patterning and growth. We identify early differences in the patterning of cover and ground scales on the young wing and quantify geometrical parameters of growing scale features, which suggest that surface growth is critical to structure formation. Our quantitative, time-resolved in vivo imaging of butterfly scale development provides the foundation for decoding the processes and biomechanical principles involved in the formation of functional structures in biological materials.

[Ultrastructure observations on antennal sensilla of Heortia vitessoides the most serious pest of Aquilaria sinensis].

Heortia vitessoides is the most serious pest of Aquilaria sinensis,which is an economically important evergreen tree native to China and is the principal source of Chinese agarwood. In severe infestations,the insects completely eat up the leaves of A. sinensis,causing severe economic losses. In a more recent study,we found that the antennal sensilla of adult play important roles in the host location,mating and oviposition of H. vitessoides. Here,the external morphology of the antennal sensilla of H. vitessoides were examined using scanning electron microscopy. The result showed that the antennae of both sexes of H. vitessoides were filiform in shape,which consist of the scape,pedicel and about 64 segments of flagellomeres. Eight morphological sensilla types were recorded in both sexes,including sensilla trichodea,sensilla chaetica,sensilla basiconica,sensilla coeloconica,sensilla styloconica,sensilla auricillica,sensilla squamiformia and böhm bristle. Major differences were recorded in the distribution and quantity of different sensilla types in each segment of antenna. The sensillas are almost confined to the ventral and lateral surfaces rather than the back side of antennae. Antennal flagella contained the most sensilla while the scape and pedicel segments only contained böhm bristles and sensilla squamiformias. Sensilla trichodea Ⅲ were only found on male antennae. These results are discussed in relation to the possible roles of the sensilla types in the host location,mating and oviposition selection behavior of H. vitessoides.

Squamocin induce histological and ultrastructural changes in the midgut cells of Anticarsia gemmatalis (Lepidoptera: Noctuidae).

Annonaceous acetogenins (Annona squamosa Linnaeus) comprises of a series of natural products which are extracted from Annonaceae species, squamocin proved to be highly efficient among those agents. Squamocin is mostly referred as a lethal agent for midgut cells of different insects, with toxic effects when tested against larva of some insects. In present study, LC50 and LC90 of squamocin for A. gemmatalis Hübner (Lepidoptera: Noctuidae) were calculated using probit analysis. Morphological changes in midgut cells were analyzed under light, fluorescence and transmission electron microscopes when larvae were treated with LC50 and LC90 of squamocin for 24, 48 and 72 h. Results revealed that the maximum damage to midgut cells was found under LC90 where it showed digestive cells with enlarged basal labyrinth, highly vacuolated cytoplasm, damaged apical surface, cell protrusions to the gut lumen, autophagy and cell death. The midgut goblet cells showed a strong disorganization of their microvilli. Likewise, in insects treated with squamocin, mitochondria were not marked with Mitotracker fluorescent probe, suggesting some molecular damage in these organelles, which was reinforced by decrease in the respiration rate in these insects. These results demonstrate that squamocin has potential to induce enough morphological changes in midgut through epithelial cell damage in A. gemmatalis.

Digestive organ in the female reproductive tract borrows genes from multiple organ systems to adopt critical functions.

Persistent adaptive challenges are often met with the evolution of novel physiological traits. Although there are specific examples of single genes providing new physiological functions, studies on the origin of complex organ functions are lacking. One such derived set of complex functions is found in the Lepidopteran bursa copulatrix, an organ within the female reproductive tract that digests nutrients from the male ejaculate or spermatophore. Here, we characterized bursa physiology and the evolutionary mechanisms by which it was equipped with digestive and absorptive functionality. By studying the transcriptome of the bursa and eight other tissues, we revealed a suite of highly expressed and secreted gene products providing the bursa with a combination of stomach-like traits for mechanical and enzymatic digestion of the male spermatophore. By subsequently placing these bursa genes in an evolutionary framework, we found that the vast majority of their novel digestive functions were co-opted by borrowing genes that continue to be expressed in nonreproductive tissues. However, a number of bursa-specific genes have also arisen, some of which represent unique gene families restricted to Lepidoptera and may provide novel bursa-specific functions. This pattern of promiscuous gene borrowing and relatively infrequent evolution of tissue-specific duplicates stands in contrast to studies of the evolution of novelty via single gene co-option. Our results suggest that the evolution of complex organ-level phenotypes may often be enabled (and subsequently constrained) by changes in tissue specificity that allow expression of existing genes in novel contexts, such as reproduction. The extent to which the selective pressures encountered in these novel roles require resolution via duplication and sub/neofunctionalization is likely to be determined by the need for specialized reproductive functionality. Thus, complex physiological phenotypes such as that found in the bursa offer important opportunities for understanding the relative role of pleiotropy and specialization in adaptive evolution.

Evolution of larval life mode of Oecophoridae (Lepidoptera: Gelechioidea) inferred from molecular phylogeny.

Phylogenetic relationships within family Oecophoridae have been poorly understood. Consequently the subfamily and genus level classifications with this family problematic. A comprehensive phylogenetic analysis of Oecophoridae, the concealer moths, was performed based on analysis of 4444 base pairs of mitochondrial COI, nuclear ribosomal RNA genes (18S and 28S) and nuclear protein coding genes (IDH, MDH, Rps5, EF1a and wingless) for 82 taxa. Data were analyzed using maximum likelihood (ML), parsimony (MP) and Bayesian (BP) phylogenetic frameworks. Phylogenetic analyses indicated that (i) genera Casmara, Tyrolimnas and Pseudodoxia did not belong to Oecophoridae, suggesting that Oecophoridae s. authors was not monophyletic; (ii) other oecophorids comprising two subfamilies, Pleurotinae and Oecophorinae, were nested within the same clade, and (iii) Martyringa, Acryptolechia and Periacmini were clustered with core Xyloryctidae. They appeared to be sister lineage with core Oecophoridae. BayesTraits were implemented to explore the ancestral character states to infer historical microhabitat patterns and sheltering strategy of larvae. Reconstruction of ancestral microhabitat of oecophorids indicated that oecophorids might have evolved from dried plant feeders and further convergently specialized. The ancestral larva sheltering strategy of oecophorids might have used a silk tube by making itself, shifting from mining leaves.

A new species of the genus Sovia (Lepidoptera: Hesperiidae) from Qinling-Daba Mountains of China.

In this paper, a new species Sovia lii spec. nov. is described from Qinling-Daba Mountains in S. Gansu province and S. Shaanxi province, W. China. Variability of wing pattern, differences with its similar congeners and some biological information of this new taxon are introduced. External variability of Sovia lucasii, which is discovered from Shaanxi for the first time and is sympatric with the new species there, is illustrated and discussed. A brief analysis of the distributional pattern of the genus Sovia is provided. A key to the genus is given.

Embryogenesis of Heliconius erato (Lepidoptera, Nymphalidae): a contribution to the anatomical development of an evo-devo model organism.

This study reports on the embryogenesis of Heliconius erato phyllis between blastoderm formation and the prehatching larval stage. Syncytial blastoderm formation occurred approximately 2 h after egg laying (AEL) and at about 4 h, the cellular blastoderm was formed. The germ band arose from the entire length of the blastoderm, and rapidly became compacted occupying approximately two-thirds of the egg length. At about 7 h AEL, protocephalon and protocorm differentiation occurred. Continued proliferation of the germ band was followed by penetration into the yolk mass, forming a C-shaped embryo at about 10 h. Approximately 12 h AEL, the gnathal, thoracic and abdominal segments became visible. The primordium of the mouthparts and thoracic legs formed as paired evaginations, while the prolegs formed as paired lobes. At about 30 h, the embryo reversed dorsoventrally. Approximately 32 h AEL, the protocephalon and gnathal segments fused, shifting the relative position of the rudimentary appendages in this region. At about 52 h, the embryo was U-shaped in lateral view and at approximately 56 h, the bristles began evagination from the larval cuticle. Larvae hatched at about 72 h. We found that H. erato phyllis followed an embryonic pattern consistent with long-germ embryogenesis. Thus, we believe that H. erato phyllis should be classified as a long-germ lepidopteran. The study of H. erato phyllis embryogenesis provided a structural glimpse into the morphogenetic events that occur in the Heliconius egg period. This study could help future molecular approaches to understanding the evolution of Heliconius development.

Morphology-based phylogeny and biogeography of Wockia (Lepidoptera: Urodidae) with description of a new species from Japan and South Korea.

Wockia, one of the six genera within the lepidopteran family Urodidae, currently includes nine species distributed in Holarctic, Oriental, and Neotropical regions. A new species of the genus, W. magna sp. nov., is described from Japan and South Korea. This is the first record of Urodidae from Japan. A cladistic analysis was constructed based on 27 morphological characters from five ingroup species and two outgroup taxa. A single most parsimonious tree was found (length = 38, CI = 71, RI = 70). The resulting tree failed to recover the monophyly of Wockia chewbacca and a clade Wockia sensu stricto, including four congeners from North America, temperate East Asia and Southeast Asia. Wockia sensu stricto was resolved as monophyletic and was divided into two subgroups, one including W. asperipunctella and W. magna, and the other including W. koreana and W. variata. The sister-group relationship of W. asperipunctella and W. magna was moderately supported. Synapomorphies of Wockia sensu stricto are provided from the resulting phylogeny. Systematic definitions of Wockia and other allied genera are revised. Optimal ancestral area reconstruction implemented in DIVA resulted in different hypotheses for Wockia, depending on how to constrain the maximum number of areas. The maximum area number set as two yielded a more likely scenario suggesting that Wockia sensu stricto originated in temperate East Asia and then dispersed into the Oriental region, North America, and Europe. A working hypothesis and other equally possible alternative explanations for the biogeography of Wockia are provided.

Critical electrolyte concentration of silk gland chromatin of the sugarcane borer Diatraea saccharalis, induced using agrochemicals.

The sugarcane borer Diatraea saccharalis is widely known as the main pest of sugarcane crop, causing increased damage to the entire fields. Measures to control this pest involve the use of chemicals and biological control with Cotesia flavipes wasps. In this study, we evaluated the insecticides fipronil (Frontline; 0.0025%), malathion (Malatol Bio Carb; 0.4%), cipermetrina (Galgotrin; 10%), and neem oil (Natuneem; 100%) and the herbicide nicosulfuron (Sanson 40 SC; 100%) in the posterior region silk glands of 3rd- and 5th-instar D. saccharalis by studying the variation in the critical electrolyte concentration (CEC). Observations of 3rd-instar larvae indicated that malathion, cipermetrina, and neem oil induced increased chromatin condensation that may consequently disable genes. Tests with fipronil showed no alteration in chromatin condensation. With the use of nicosulfuron, there was chromatin and probable gene decompaction. In the 5th-instar larvae, the larval CEC values indicated that malathion and neem oil induced increased chromatin condensation. The CEC values for 5th-instar larvae using cipermetrina, fipronil, and nicosulfuron indicated chromatin unpacking. These observations led us to conclude that the quantity of the pesticide does not affect the mortality of these pests, can change the conformation of complexes of DNA, RNA, and protein from the posterior region of silk gland cells of D. saccharalis, activating or repressing the expression of genes related to the defense mechanism of the insect and contributing to the selection and survival of resistant individuals.

A new species of Aricoris Westwood, [1851] belonging to "chilensis" group (Lepidoptera: Riodinidae).

A new species of Riodinidae from the grasslands of South Brazil, Paraguay and Argentina, Aricoris schneideri sp. nov., is described.

A survey of Eupithecia Curtis, 1825 (Lepidoptera, Geometridae, Larentiinae) in Mongolia with descriptions of two new species.

The Eupithecia fauna of Mongolia is surveyed in this paper, drawing on previous publications and newly collected material. In total 57 species are recorded. Two species Eupithecia anikini Mironov & Galsworthy, sp. n. and Eupithecia munguata Mironov & Galsworthy, sp. n. are described as new. Twelve species are recorded as new for the fauna of Mongolia, E. olgae, E. illaborata, E. bastelbergeri, E. pusillata, E. actaeata, E. vicariata, E. saisanaria, E. simpliciata, E. impolita, E. assimilata, E. thalictrata and E. exactata. Habitus and genitalia are illustrated for new species, and for little known older species where illustrations are not available in recent papers. A further 2 previously described species are synonymised, E. mongolica Vojnits of E. satyrata Hübner, and E. procera Vojnits of E. exactata Staudinger.

On the identity of Ypsolopha lonicerella Stöckl, 1922 (Lepidoptera, Ypsolophidae).

Ypsolopha lonicerella Stöckl, 1922 is shown to be a junior synonym of Y. leuconotella Snellen, 1884.

A taxonomic revision of the genus Edosa Walker, 1886 from China (Lepidoptera, Tineidae, Perissomasticinae).

The genus Edosa Walker, 1886 is revised in China. Of the 31 recognized species, 23 are described as new: Edosa aurea sp. nov., E. baculiformis sp. nov., E. bicolor sp. nov., E. bifurcata sp. nov., E. carinata sp. nov., E. conchata sp. nov., E. cornuta sp. nov., E. curvidorsalis sp. nov., E. dentata sp. nov., E. digitata sp. nov., E. duoprojecta sp. nov., E. elongata sp. nov., E. eminens sp. nov., E. hamata sp. nov., E. longicornis sp. nov., E. minuta sp. nov., E. robinsoni sp. nov., E. robustispina sp. nov., E. semicircularis sp. nov., E. torta sp. nov., E. truncatula sp. nov., E. uncusella sp. nov., and E. varians sp. nov.; Edosa gaedikei nom. nov. is proposed as a replacement name for Edosa spinosa Gaedike, 2012, a junior homonym of Edosa spinosa Gaedike, 1984; four species are newly recorded for China: E. hendrixella Robinson, 2008, E. crayella Robinson, 2008, E. orphnodes (Meyrick, 1911) and E. smithaella Robinson, 2008; the female of E. hendrixella is described for the first time. Phylogenetic relationships of the Chinese species, except E. malthacopis (Meyrick, 1936), are postulated based on 42 morphological characters. The phylogenetic analysis of the morphology matrix yielded one most parsimonious tree (length 121 steps, CI = 0.56, RI = 0.81), based on which seven species-groups are proposed: the hemichrysella-group, the subochraceella-group, the pyrochra-group, the duoprojecta-group, the eminens-group, the crayella-group and the uncusella-group. The whole body structures of the genus are presented and photographs of adults and genitalia are provided, along with a key to all the known Chinese species and maps to show the distribution of Edosa in China. A world checklist of Edosa is given as an appendix, in which 41 new combinations are included.

Review of the Olethreutes bowmanana-magadana species group, with the description of a new species from the Amur River, Russia (Lepidoptera, Tortricidae).

We define and review the Olethreutes bowmanana-magadana species group comprised of Olethreutes bowmanana (McDunnough), O. magadana (Falkovitsh), O. kamtshadala (Falkovitsh), and O. pivanica, sp. n., the last from petrophytous sunny slopes in the Amur River. The new species differs from other species of the group by a narrow, triangular-shaped apical process from the sacculus; a round, wide, apical bulb from the cucullus; and a longer, narrow neck of the valva.

A new species of Pheles Herrich-Schaeffer from Northeast Brazil (Lepidoptera, Riodinidae).

A new species of Riodinidae, Pheles caatingensis Callaghan & Nobre, sp. nov. from Ceará and Pernambuco State in Brazil is described, along with its habitat, behavior and taxonomic differences with other members of the genus Pheles. Notes are included on distribution and mimetic relationships with other sympatric insects.

Two new species of Esthemopsis C. Felder & R. Felder, 1865 (Lepidoptera: Riodinidae: Symmachiini) from southeastern and northeastern Brazil, with taxonomic comments on Esthemopsis teras (Stichel, 1910) stat. rev. and Esthemopsis pallida Lathy, 1932 stat. nov.

Two new species of Esthemopsis C. Felder & R. Felder, 1865 from southeastern and northeastern Brazil are described, Esthemopsis sateri sp. nov. and Esthemopsis diamantina sp. nov., and revised and new combinations are proposed for Esthemopsis teras (Stichel, 1910) stat. rev. and Esthemopsis pallida Lathy, 1932 stat. nov., respectively. Lectotypes are designated for Lepricornis teras Stichel, 1910 and Esthemopsis caerulea pallida Lathy, 1932. The habitus of male and female specimens of the species discussed are illustrated; male head and labial palpus pictures, male and female genitalia illustrations are provided.

Discovery of a third species of Lamproptera Gray, 1832 (Lepidoptera: Papilionidae).

A newly discovered, third species of the genus Lamproptera (Lepidoptera: Papilionidae) is described, 183 years after the second currently recognised species was first named. Lamproptera paracurius Hu, Zhang & Cotton sp. n., from N.E. Yunnan, China, is based on marked differences in external morphology and male genital structure. The species is confirmed as a member of the genus, and detailed comparisons are made with other taxa included in the genus. Keys to Lamproptera species based on external characters and male genitalia are included.

Sattleria revisited: unexpected cryptic diversity on the Balkan Peninsula and in the south-eastern Alps (Lepidoptera: Gelechiidae).

The taxonomy of Sattleria Povolný from the high mountain systems on the Balkan Peninsula and the adjacent parts of the Alps (south-eastern Alps, Dinaric Alps, Rila Mountains) is revised based on recently collected material and re-examined museum vouchers. Adult morphology and molecular data of the COI barcode region support the existence of six strictly allopatric species in this area, including four new species: Sattleria sophiae Timossi, sp. nov. (Parco Paneveggio-Pale di San Martino, Dolomites, Prov. Trento, Italy), Sattleria dolomitica Huemer, sp. nov. (Eastern Dolomites, Prov. South Tyrol, Italy), Sattleria dinarica Huemer, sp. nov. (Durmitor NP, Dinaric Alps, Montenegro) and Sattleria haemusi Huemer, sp. nov. (Rila Mts., Bulgaria; Sar Planina, Macedonia). 

A tropical horde of counterfeit predator eyes.

We propose that the many different, but essentially similar, eye-like and face-like color patterns displayed by hundreds of species of tropical caterpillars and pupae-26 examples of which are displayed here from the dry, cloud, and rain forests of Area de Conservacion Guanacaste (ACG) in northwestern Costa Rica-constitute a huge and pervasive mimicry complex that is evolutionarily generated and sustained by the survival behavior of a large and multispecific array of potential predators: the insect-eating birds. We propose that these predators are variously and innately programmed to flee when abruptly confronted, at close range, with what appears to be an eye of one of their predators. Such a mimetic complex differs from various classical Batesian and Müllerian mimicry complexes of adult butterflies in that (i) the predators sustain it for the most part by innate traits rather than by avoidance behavior learned through disagreeable experiences, (ii) the more or less harmless, sessile, and largely edible mimics vastly outnumber the models, and (iii) there is no particular selection for the eye-like color pattern to closely mimic the eye or face of any particular predator of the insect-eating birds or that of any other member of this mimicry complex. Indeed, selection may not favor exact resemblance among these mimics at all. Such convergence through selection could create a superabundance of one particular false eyespot or face pattern, thereby increasing the likelihood of a bird species or guild learning to associate that pattern with harmless prey.

Evaluation of cellulolytic activity in insect digestive fluids.

Efficient and low-cost cellulolytic enzymes are urgently needed to degrade recalcitrant plant biomass during the industrial production of lignocellulosic biofuels. Here, the cellulolytic activities in the gut fluids of 54 insect species that belong to 7 different taxonomic orders were determined using 2 different substrates, carboxymethyl cellulose (CMC) (approximating endo-ß-1,4-glucanase) and filter paper (FP) (total cellulolytic activities). The use of CMC as the substrate in the zymogram analysis resulted in the detection of distinct cellulolytic protein bands. The cellulolytic activities in the digestive system of all the collected samples were detected using cellulolytic activity analysis. The highest CMC gut fluid activities were found in Coleoptera and Orthoptera, while FP analysis indicated that higher gut fluid activities were found in several species of Coleoptera and Lepidoptera. In most cases, gut fluid activities were higher with CMC than with FP substrate, except for individual Lepidoptera species. Our data indicate that the origin of cellulolytic enzymes probably reflects the phylogenetic relationship and feeding strategies of different insects.