New ways to acquire resistance: imperfect convergence in insect adaptations to a potent plant toxin.

Evolution of insensitivity to the toxic effects of cardiac glycosides has become a model in the study of convergent evolution, as five taxonomic orders of insects use the same few similar amino acid substitutions in the otherwise highly conserved Na,K-ATPase α. We show here that insensitivity in pyrgomorphid grasshoppers evolved along a slightly divergent path. As in other lineages, duplication of the Na,K-ATPase α gene paved the way for subfunctionalization: one copy maintains the ancestral, sensitive state, while the other copy is resistant. Nonetheless, in contrast with all other investigated insects, the grasshoppers' resistant copy shows length variation by two amino acids in the first extracellular loop, the main part of the cardiac glycoside-binding pocket. RT-qPCR analyses confirmed that this copy is predominantly expressed in tissues exposed to the toxins, while the ancestral copy predominates in the nervous tissue. Functional tests with genetically engineered Drosophila Na,K-ATPases bearing the first extracellular loop of the pyrgomorphid genes showed the derived form to be highly resistant, while the ancestral state is sensitive. Thus, we report convergence in gene duplication and in the gene targets for toxin insensitivity; however, the means to the phenotypic end have been novel in pyrgomorphid grasshoppers.

No signal of deleterious mutation accumulation in conserved gene sequences of extant asexual hexapods.

Loss of sex and recombination is generally assumed to impede the effectiveness of purifying selection and to result in the accumulation of slightly deleterious mutations. Empirical evidence for this has come from several studies investigating mutational load in a small number of individual genes. However, recent whole transcriptome based studies have yielded inconsistent results, hence questioning the validity of the assumption of mutational meltdown in asexual populations. Here, we study the effectiveness of purifying selection in eight asexual hexapod lineages and their sexual relatives, as present in the 1 K Insect Transcriptome Evolution (1KITE) project, covering eight hexapod groups. We analyse the accumulation of slightly deleterious nonsynonymous and synonymous point mutations in 99 single copy orthologue protein-coding loci shared among the investigated taxa. While accumulation rates of nonsynonymous mutations differed between genes and hexapod groups, we found no effect of reproductive mode on the effectiveness of purifying selection acting at nonsynonymous and synonymous sites. Although the setup of this study does not fully rule out nondetection of subtle effects, our data does not support the established consensus of asexual lineages undergoing 'mutational meltdown'.

Evolutionary history of Polyneoptera and its implications for our understanding of early winged insects.

Polyneoptera represents one of the major lineages of winged insects, comprising around 40,000 extant species in 10 traditional orders, including grasshoppers, roaches, and stoneflies. Many important aspects of polyneopteran evolution, such as their phylogenetic relationships, changes in their external appearance, their habitat preferences, and social behavior, are unresolved and are a major enigma in entomology. These ambiguities also have direct consequences for our understanding of the evolution of winged insects in general; for example, with respect to the ancestral habitats of adults and juveniles. We addressed these issues with a large-scale phylogenomic analysis and used the reconstructed phylogenetic relationships to trace the evolution of 112 characters associated with the external appearance and the lifestyle of winged insects. Our inferences suggest that the last common ancestors of Polyneoptera and of the winged insects were terrestrial throughout their lives, implying that wings did not evolve in an aquatic environment. The appearance of the first polyneopteran insect was mainly characterized by ancestral traits such as long segmented abdominal appendages and biting mouthparts held below the head capsule. This ancestor lived in association with the ground, which led to various specializations including hardened forewings and unique tarsal attachment structures. However, within Polyneoptera, several groups switched separately to a life on plants. In contrast to a previous hypothesis, we found that social behavior was not part of the polyneopteran ground plan. In other traits, such as the biting mouthparts, Polyneoptera shows a high degree of evolutionary conservatism unique among the major lineages of winged insects.

Synthesis of rare-earth metal and rare-earth metal-fluoride nanoparticles in ionic liquids and propylene carbonate.

Decomposition of rare-earth tris(N,N'-diisopropyl-2-methylamidinato)metal(III) complexes [RE{MeC(N(iPr)2)}3] (RE(amd)3; RE = Pr(III), Gd(III), Er(III)) and tris(2,2,6,6-tetramethyl-3,5-heptanedionato)europium(III) (Eu(dpm)3) induced by microwave heating in the ionic liquids (ILs) 1-butyl-3-methylimidazolium tetrafluoroborate ([BMIm][BF4]), 1-butyl-3-methylimidazolium bis(trifluoromethylsulfonyl)imide ([BMIm][NTf2]) and in propylene carbonate (PC) yield oxide-free rare-earth metal nanoparticles (RE-NPs) in [BMIm][NTf2] and PC for RE = Pr, Gd and Er or rare-earth metal-fluoride nanoparticles (REF3-NPs) in the fluoride-donating IL [BMIm][BF4] for RE = Pr, Eu, Gd and Er. The crystalline phases and the absence of significant oxide impurities in RE-NPs and REF3-NPs were verified by powder X-ray diffraction (PXRD), selected area electron diffraction (SAED) and high-resolution X-ray photoelectron spectroscopy (XPS). The size distributions of the nanoparticles were determined by transmission electron microscopy (TEM) and high-angle annular dark-field scanning transmission electron microscopy (HAADF-STEM) to an average diameter of (11 ± 6) to (38 ± 17) nm for the REF3-NPs from [BMIm][BF4]. The RE-NPs from [BMIm][NTf2] or PC showed diameters of (1.5 ± 0.5) to (5 ± 1) nm. The characterization was completed by energy-dispersive X-ray spectroscopy (EDX).

Morphological convergences in Ameles Burmeister and Pseudoyersinia Kirby: Taxonomic implications of wing reduction and flight predisposition in some West-Mediterranean Amelini (Insecta: Mantodea).

Species in the genus Ameles Burmeister and Pseudoyersinia Kirby (Amelini) are traditionally defined as small-sized, ground-dwelling mantids whose males are distinct for being, respectively, macropterous and brachypterous. However, comparative morphological studies across Amelini confirmed the existence of short-winged males in Ameles, suggesting that this traditional diagnostic concept does not apply to all species. Our analyses of several species from West Mediterranean localities (Canary Islands, Spain, Italy, and Morocco) resulted in the relocation of Pseudoyersinia andreae Galvagni, 1976 to Ameles as Ameles andreae (Galvagni, 1976) (n. comb.) with Ameles insularis Agabiti, Ippolito Lombardo, 2010 as its new synonym (n. syn.), the clarification of the taxonomic identity of A. gracilis (Brullé, 1838) and A. maroccana Uvarov, 1931, including diagnoses of their males, and the description of Ameles spallanzania obscura (n. ssp.) (from Spain). We also take the opportunity to describe Pseudoyersinia maroccana (n. sp.) (from Morocco) based on museum specimens separating it from Ameles maroccana Uvarov, 1931. We also found that wing length is positively correlated to ocelli size. We discuss this trend from an ecological, evolutionary, and biogeographic perspectives to both facilitate species circumscription and justify the taxonomic modifications herein introduced.

Synthesis of metal-fluoride nanoparticles supported on thermally reduced graphite oxide.

Metal-fluoride nanoparticles, (MF x -NPs) with M = Fe, Co, Pr, Eu, supported on different types of thermally reduced graphite oxide (TRGO) were obtained by microwave-assisted thermal decomposition of transition-metal amidinates, (M{MeC[N(iPr)]2} n ) or [M(AMD) n ] with M = Fe(II), Co(II), Pr(III), and tris(2,2,6,6-tetramethyl-3,5-heptanedionato)europium, Eu(dpm)3, in the presence of TRGO in the ionic liquid (IL) 1-butyl-3-methylimidazolium tetrafluoroborate ([BMIm][BF4]). The crystalline phases of the metal fluorides synthesized in [BMIm][BF4] were identified by powder X-ray diffraction (PXRD) to be MF2 for M = Fe, Co and MF3 for M = Eu, Pr. The diameters and size distributions of MF x @TRGO were from (6 ± 2) to (102 ± 41) nm. Energy-dispersive X-ray spectroscopy (EDX) and X-ray photoelectron spectroscopy (XPS) were used for further characterization of the MF x -NPs. Electrochemical investigations of the FeF2-NPs@TRGO as cathode material for lithium-ion batteries were evaluated by galvanostatic charge/discharge profiles. The results indicate that the FeF2-NPs@TRGO as cathode material can present a specific capacity of 500 mAh/g at a current density of 50 mA/g, including a significant interfacial charge storage contribution. The obtained nanomaterials show a good rate capacity as well (220 mAh/g and 130 mAh/g) at a current density of 200 and 500 mA/g, respectively.

Corrigendum: Synthesis of Metal Nanoparticles and Metal Fluoride Nanoparticles from Metal Amidinate Precursors in 1-Butyl-3-Methylimidazolium Ionic Liquids and Propylene Carbonate.

[This corrects the article DOI: 10.1002/open.201600105.].

Soft, Wet-Chemical Synthesis of Metastable Superparamagnetic Hexagonal Close-Packed Nickel Nanoparticles in Different Ionic Liquids.

The microwave-induced decomposition of bis{N,N'-diisopropylacetamidinate}nickel(II) [Ni{MeC(NiPr)2 }2 ] or bis(1,5-cyclooctadiene)nickel(0) [Ni(COD)2 ] in imidazolium-, pyridinium-, or thiophenium-based ionic liquids (ILs) with different anions (tetrafluoroborate, [BF4 ]- , hexafluorophosphate, [PF6 ]- , and bis(trifluoromethylsulfonyl)imide, [NTf2 ]- ) yields small, uniform nickel nanoparticles (Ni NPs), which are stable in the absence of capping ligands (surfactants) for more than eight weeks. The soft, wet-chemical synthesis yields the metastable Ni hexagonal close-packed (hcp) and not the stable Ni face-centered cubic (fcc) phase. The size of the nickel nanoparticles increases with the molecular volume of the used anions from about 5 nm for [BF4 ]- to ≈10 nm for [NTf2 ]- (with 1-alkyl-3-methyl-imidazolium cations). The n-butyl-pyridinium, [BPy]+ , cation ILs reproducibly yield very small nickel nanoparticles of 2(±1) nm average diameter. The Ni NPs were characterized by high-resolution transmission electron microscopy (HR-TEM) and powder X-ray diffraction. An X-ray photoelectron spectroscopic (XPS) analysis shows an increase of the binding energy (EB ) of the electron from the Ni 2p3/2 orbital of the very small 2(±1) nm diameter Ni particles by about 0.3 eV to EB =853.2 eV compared with bulk Ni0 , which is traced to the small cluster size. The Ni nanoparticles show superparamagnetic behavior from 150 K up to room temperature. The saturation magnetization of a Ni (2±1 nm) sample from [BPy][NTf2 ] is 2.08 A m2 kg-1 and of a Ni (10±4 nm) sample from [LMIm][NTf2 ] it is 0.99 A m2 kg-1 , ([LMIm]=1-lauryl-3-methyl- imidazolium). The Ni NPs were active catalysts in IL dispersions for 1-hexene or benzene hydrogenation. Over 90 % conversion was reached under 5 bar H2 in 1 h at 100 °C for 1-hexene and a turnover frequency (TOF) up to 1330 molhexane (molNi )-1 h-1 or in 60 h at 100 °C for benzene hydrogenation and TOF=23 molcyclohexane (molNi )-1 h-1 .

Synthesis of Metal Nanoparticles and Metal Fluoride Nanoparticles from Metal Amidinate Precursors in 1-Butyl-3-Methylimidazolium Ionic Liquids and Propylene Carbonate.

Decomposition of transition-metal amidinates [M{MeC(NiPr)2} n ] [M(AMD) n ; M=MnII, FeII, CoII, NiII, n=2; CuI, n=1) induced by microwave heating in the ionic liquids (ILs) 1-butyl-3-methylimidazolium tetrafluoroborate ([BMIm][BF4]), 1-butyl-3-methylimidazolium hexafluorophosphate ([BMIm][PF6]), 1-butyl-3-methylimidazolium trifluoromethanesulfonate (triflate) ([BMIm][TfO]), and 1-butyl-3-methylimidazolium tosylate ([BMIm][Tos]) or in propylene carbonate (PC) gives transition-metal nanoparticles (M-NPs) in non-fluorous media (e.g. [BMIm][Tos] and PC) or metal fluoride nanoparticles (MF2-NPs) for M=Mn, Fe, and Co in [BMIm][BF4]. FeF2-NPs can be prepared upon Fe(AMD)2 decomposition in [BMIm][BF4], [BMIm][PF6], and [BMIm][TfO]. The nanoparticles are stable in the absence of capping ligands (surfactants) for more than 6 weeks. The crystalline phases of the metal or metal fluoride synthesized in [BMIm][BF4] were identified by powder X-ray diffraction (PXRD) to exclusively Ni- and Cu-NPs or to solely MF2-NPs for M=Mn, Fe, and Co. The size and size dispersion of the nanoparticles were determined by transmission electron microscopy (TEM) to an average diameter of 2(±2) to 14(±4) nm for the M-NPs, except for the Cu-NPs in PC, which were 51(±8) nm. The MF2-NPs from [BMIm][BF4] were 15(±4) to 65(±18) nm. The average diameter from TEM is in fair agreement with the size evaluated from PXRD with the Scherrer equation. The characterization was complemented by energy-dispersive X-ray spectroscopy (EDX). Electrochemical investigations of the CoF2-NPs as cathode materials for lithium-ion batteries were simply evaluated by galvanostatic charge/discharge profiles, and the results indicated that the reversible capacity of the CoF2-NPs was much lower than the theoretical value, which may have originated from the complex conversion reaction mechanism and residue on the surface of the nanoparticles.

Six, not two, species of Acisoma pintail dragonfly (Odonata: Libellulidae).

The dragonfly genus Acisoma is revised based on adult male morphology and COI sequence data. Six species are recognised, including the new species A. attenboroughi sp. nov. Diagnoses and a key to males of all species and illustrations of all relevant characters are provided. A. inflatum, A. variegatum and A. trifidum are confined to continental Africa, while A. panorpoides is restricted to Asia. A. ascalaphoides is known only from threatened littoral forest fragments on the east coast of Madagascar, while A. attenboroughi is widespread across the island. The new species honours Sir David Attenborough on his 90th birthday.

A review of the research on Canary Islands praying mantises (Mantodea).

For many years researchers studying the fauna of the Canary Islands have only sparsely treated the charismatic insect order of praying mantises (Mantodea). By studying the known literature it becomes obvious that there are several inconsistencies regarding mantodean taxonomy as well as the number of actual species and their distribution within the archipelago. In the present contribution, the literature treating the Canary Island Mantodea fauna is thoroughly reviewed, and the distribution of the ten known Mantodea species is presented with additional comments on the taxonomic problems. The taxonomy and distribution of the Canary Island Amelinae has been causing some confusion in the literature and is therefore discussed for a better understanding of the composition of this group. So far, seven species of Amelinae (two species of Ameles Burmeister and five species of Pseudoyersinia Kirby) are recognized for the Canary Islands. A careful survey of the taxonomic literature indicates that there is a substantial degree of variability in this group, which may have led to frequent misidentifications. This is reflected by the inconsistent literature data on the distribution of several Pseudoyersinia species within the archipelago. A crucial amendment and clarification is provided regarding the date of description (1838) of Ameles gracilis (Brullé) and Ameles limbata (Brullé) as it varies in the literature. Nomenclatural changes regarding the homonymy of Mantis limbata Brullé and Mantis limbata Hahn, previously proposed by Koçak & Kemal (2008), are rejected and discussed in detail. One of the Mantodea species, Empusa pennata (Thunberg), is frequently mentioned in the literature to be present on the Canary Islands, but after carefully reviewing the literature data it is evident that this species has never actually been found on any of the islands. The aim of this publication is to lay a basis for future research on the Mantodea fauna of the Canary Islands, their phylogenetic placement and history of dispersal. This is even more important as the archipelago is a European endemism hot spot for praying mantises and these basic parameters are crucial for implementing plans regarding conservation strategies on the islands.

Colloidal nickel/gallium nanoalloys obtained from organometallic precursors in conventional organic solvents and in ionic liquids: noble-metal-free alkyne semihydrogenation catalysts.

Efforts to replace noble-metal catalysts by low-cost alternatives are of constant interest. The organometallic, non-aqueous wet-chemical synthesis of various hitherto unknown nanocrystalline Ni/Ga intermetallic materials and the use of NiGa for the selective semihydrogenation of alkynes to alkenes are reported. Thermal co-hydrogenolysis of the all-hydrocarbon precursors [Ni(COD)2] (COD = 1,5-cyclooctadiene) and GaCp* (Cp* = pentamethylcyclopentadienyl) in high-boiling organic solvents mesitylene and n-decane in molar ratios of 1 : 1, 2 : 3 and 3 : 1 yields the nano-crystalline powder materials of the over-all compositions NiGa, Ni2Ga3 and Ni3Ga, respectively. Microwave induced co-pyrolysis of the same precursors without additional hydrogen in the ionic liquid [BMIm][BF4] (BMIm = 1-butyl-3-methyl-imidazolium) selectively yields the intermetallic phases NiGa and Ni3Ga from the respective 1 : 1 and 3 : 1 molar ratios of the precursors. The obtained materials are characterized by transmission electron microscopy (TEM), energy dispersive X-ray spectroscopy (EDX), IR, powder X-ray diffraction (PXRD) and atomic absorption spectroscopy (AAS). The single-source precursor [Ni(GaCp*)(PMe3)3] with a fixed Ni : Ga stoichiometry of 1 : 1 was employed as well. In comparison with the co-hydrogenolytic dual precursor source approach it turned out to be less practical due to inefficient nickel incorporation caused by the parasitic formation of stable [Ni(PMe3)4]. The use of ionic liquid [BMIm][BF4] as a non-conventional solvent to control the reaction and stabilize the nanoparticles proved to be particularly advantageous and stable colloids of the nanoalloys NiGa and Ni3Ga were obtained. A phase-selective Ni/Ga colloid synthesis in conventional solvents and in the presence of surfactants such as hexadecylamine (HDA) was not feasible due to the undesired reactivity of HDA with GaCp* leading to inefficient gallium incorporation. Recyclable NiGa nanoparticles selectively semihydrogenate 1-octyne and diphenylacetylene (tolan) to 1-octene and diphenylethylene, respectively, with a yield of about 90% and selectivities of up to 94 and 87%. Ni-NPs yield alkanes with a selectivity of 97 or 78%, respectively, under the same conditions.

Viability characterization of Taxus chinensis plant cell suspension cultures by rapid colorimetric- and image analysis-based techniques.

For the commercially established process of paclitaxel production with Taxus chinensis plant cell culture, the size of plant cell aggregates and phenotypic changes in coloration during cultivation have long been acknowledged as intangible parameters. So far, the variability of aggregates and coloration of cells are challenging parameters for any viability assay. The aim of this study was to investigate simple and non-toxic methods for viability determination of Taxus cultures in order to provide a practicable, rapid, robust and reproducible way to sample large amounts of material. A further goal was to examine whether Taxus aggregate cell coloration is related to general cell viability and might be exploited by microscopy and image analysis to gain easy access to general cell viability. The Alamar Blue assay was found to be exceptionally eligible for viability estimation. Moreover, aggregate coloration, as a morphologic attribute, was quantified by image analysis and found to be a good and traceable indicator of T. chinensis viability.

Synthesis of Cu, Zn and Cu/Zn brass alloy nanoparticles from metal amidinate precursors in ionic liquids or propylene carbonate with relevance to methanol synthesis.

Microwave-induced decomposition of the transition metal amidinates {[Me(C(N(i)Pr)2)]Cu}2 (1) and [Me(C(N(i)Pr)2)]2Zn (2) in the ionic liquid 1-butyl-3-methylimidazolium tetrafluoroborate ([BMIm][BF4]) or in propylene carbonate (PC) gives copper and zinc nanoparticles which are stable in the absence of capping ligands (surfactants) for more than six weeks. Co-decomposition of 1 and 2 yields the intermetallic nano-brass phases ß-CuZn and γ-Cu3Zn depending on the chosen molar ratios of the precursors. Nanoparticles were characterized by high-angle annular dark field-scanning transmission electron microscopy (HAADF-STEM), dynamic light scattering and powder X-ray diffractometry. Microstructure characterizations were complemented by STEM with spatially resolved energy-dispersive X-ray spectrometry and X-ray photoelectron spectroscopy. Synthesis in ILs yields significantly smaller nanoparticles than in PC. ß-CuZn alloy nanoparticles are precursors to catalysts for methanol synthesis from the synthesis gas H2/CO/CO2 with a productivity of 10.7 mol(MeOH) (kg(Cu) h)(-1).

Insect phylogenomics: results, problems and the impact of matrix composition.

In this study, we investigated the relationships among insect orders with a main focus on Polyneoptera (lower Neoptera: roaches, mantids, earwigs, grasshoppers, etc.), and Paraneoptera (thrips, lice, bugs in the wide sense). The relationships between and within these groups of insects are difficult to resolve because only few informative molecular and morphological characters are available. Here, we provide the first phylogenomic expressed sequence tags data ('EST': short sub-sequences from a c(opy) DNA sequence encoding for proteins) for stick insects (Phasmatodea) and webspinners (Embioptera) to complete published EST data. As recent EST datasets are characterized by a heterogeneous distribution of available genes across taxa, we use different rationales to optimize the data matrix composition. Our results suggest a monophyletic origin of Polyneoptera and Eumetabola (Paraneoptera + Holometabola). However, we identified artefacts of tree reconstruction (human louse Pediculus humanus assigned to Odonata (damselflies and dragonflies) or Holometabola (insects with a complete metamorphosis); mayfly genus Baetis nested within Neoptera), which were most probably rooted in a data matrix composition bias due to the inclusion of sequence data of entire proteomes. Until entire proteomes are available for each species in phylogenomic analyses, this potential pitfall should be carefully considered.

The taming of an impossible child: a standardized all-in approach to the phylogeny of Hymenoptera using public database sequences.

BACKGROUND: Enormous molecular sequence data have been accumulated over the past several years and are still exponentially growing with the use of faster and cheaper sequencing techniques. There is high and widespread interest in using these data for phylogenetic analyses. However, the amount of data that one can retrieve from public sequence repositories is virtually impossible to tame without dedicated software that automates processes. Here we present a novel bioinformatics pipeline for downloading, formatting, filtering and analyzing public sequence data deposited in GenBank. It combines some well-established programs with numerous newly developed software tools (available at http://software.zfmk.de/). RESULTS: We used the bioinformatics pipeline to investigate the phylogeny of the megadiverse insect order Hymenoptera (sawflies, bees, wasps and ants) by retrieving and processing more than 120,000 sequences and by selecting subsets under the criteria of compositional homogeneity and defined levels of density and overlap. Tree reconstruction was done with a partitioned maximum likelihood analysis from a supermatrix with more than 80,000 sites and more than 1,100 species. In the inferred tree, consistent with previous studies, "Symphyta" is paraphyletic. Within Apocrita, our analysis suggests a topology of Stephanoidea + (Ichneumonoidea + (Proctotrupomorpha + (Evanioidea + Aculeata))). Despite the huge amount of data, we identified several persistent problems in the Hymenoptera tree. Data coverage is still extremely low, and additional data have to be collected to reliably infer the phylogeny of Hymenoptera. CONCLUSIONS: While we applied our bioinformatics pipeline to Hymenoptera, we designed the approach to be as general as possible. With this pipeline, it is possible to produce phylogenetic trees for any taxonomic group and to monitor new data and tree robustness in a taxon of interest. It therefore has great potential to meet the challenges of the phylogenomic era and to deepen our understanding of the tree of life.