Multiple Origins of Bioluminescence in Beetles and Evolution of Luciferase Function.

Bioluminescence in beetles has long fascinated biologists, with diverse applications in biotechnology. To date, however, our understanding of its evolutionary origin and functional variation mechanisms remains poor. To address these questions, we obtained high-quality reference genomes of luminous and nonluminous beetles in 6 Elateroidea families. We then reconstructed a robust phylogenetic relationship for all luminous families and related nonluminous families. Comparative genomic analyses and biochemical functional experiments suggested that gene evolution within Elateroidea played a crucial role in the origin of bioluminescence, with multiple parallel origins observed in the luminous beetle families. While most luciferase-like proteins exhibited a conserved nonluminous amino acid pattern (TLA346 to 348) in the luciferin-binding sites, luciferases in the different luminous beetle families showed divergent luminous patterns at these sites (TSA/CCA/CSA/LVA). Comparisons of the structural and enzymatic properties of ancestral, extant, and site-directed mutant luciferases further reinforced the important role of these sites in the trade-off between acyl-CoA synthetase and luciferase activities. Furthermore, the evolution of bioluminescent color demonstrated a tendency toward hypsochromic shifts and variations among the luminous families. Taken together, our results revealed multiple parallel origins of bioluminescence and functional divergence within the beetle bioluminescent system.

Two new species of the Longitarsusviolentus group from China (Coleoptera, Chrysomelidae, Galerucinae, Alticini).

Two new species of Longitarsus Latreille, 1829 from China are described: L.pekingensis Liang, Konstantinov & Ge, sp. nov. (Beijing) and L.xinjiangensis Liang, Konstantinov & Ge, sp. nov. (Xinjiang). Images of dorsal and lateral habitus, pronotum, head, and male and female genitalia are provided. The records of Longitarsusviolentus Weise, 1893 and Longitarsusweisei Guillebeau, 1895 in China are discussed. Holotypes of L.marguzoricus Konstantinov in Konstantinov & Lopatin, 2000 and L.violentoides Konstantinov in Konstantinov & Lopatin, 2000 are illustrated with images of pronotum and median lobe of aedeagus. A key to species of L.violentus species group is provided.

Description of three new species of Benedictus (Coleoptera, Chrysomelidae, Galerucinae, Alticini) from China, with comments on their biology and modified ethanol traps for collecting flea beetles.

The diversity and biology of the moss and leaf litter-inhabiting flea beetles are still poorly known. In this study, three new species of Benedictus are described from China: Benedictusfuanensis Ruan & Konstantinov, sp. nov., Benedictusquadrimaculatus Ruan & Konstantinov, sp. nov., and Benedictuswangi Ruan & Konstantinov, sp. nov. Comments on their biology are given. Benedictusquadrimaculatus has a highly unusual morphological feature not reported before in flea beetles: black spots on the abdominal tergites that are visible through the elytra. Traditional and modified ethanol traps were tested and proven useful for collecting leaf litter- and moss-inhabiting flea beetles. Based on our tests, eight traps could collect one specimen each day in the testing sites in Fujian Province; three traps could collect one specimen each day in the testing sites in Guangdong Province.

Characterization of the Complete Mitochondrial Genome of a Flea Beetle Luperomorpha xanthodera (Coleoptera: Chrysomelidae: Galerucinae) and Phylogenetic Analysis.

In this study, the mitochondrial genome of Luperomorpha xanthodera was assembled and annotated, which is a circular DNA molecule including 13 protein-coding genes (PCGs), 22 transfer RNA (tRNA) genes, 2 ribosomal RNA genes (12S rRNA and 16S rRNA), and 1388 bp non-coding regions (A + T rich region), measuring 16,021 bp in length. The nucleotide composition of the mitochondrial genome is 41.3% adenine (A), 38.7% thymine (T), 8.4% guanine (G), and 11.6% cytosine (C). Most of the protein-coding genes presented a typical ATN start codon (ATA, ATT, ATC, ATG), except for ND1, which showed the start codon TTG. Three-quarters of the protein-coding genes showed the complete stop codon TAR (TAA, TAG), except the genes COI, COII, ND4, and ND5, which showed incomplete stop codons (T- or TA-). All the tRNA genes have the typical clover-leaf structure, except tRNASer1 (AGN), which has a missing dihydrouridine arm (DHU). The phylogenetic results determined by both maximum likelihood and Bayesian inference methods consistently supported the monophyly of the subfamily Galerucinae and revealed that the subtribe Luperina and genus Monolepta are polyphyletic groups. Meanwhile, the classification status of the genus Luperomorpha is controversial.

Complete mitochondrial genome of Pectocera sp. (Elateridae: Dendrometrinae: Oxynopterini) and its phylogenetic implications.

The Elateridae family (click beetles) represents a highly diverse lineage that possesses a specialized clicking mechanism to startle predators. At present, however, phylogenetic relationships, especially among recognized subfamilies, remain contentious. Mitochondrial genomes (mitogenomes) can help resolve previously intractable phylogenetic relationships using morphological or limited molecular data. Here, we report the complete mitogenome of Pectocera sp. (Elateridae: Dendrometrinae: Oxynopterini), which was 15,962 bp in length and showed a typical gene number and order as most beetle mitogenomes, including 13 protein-coding genes (PCGs), 22 transfer RNA genes, 2 ribosomal RNA genes, and 1 noncoding control region (AT-rich region). Comparative genomic analyses showed a high degree of feature similarity among Pectocera sp. and other click beetles. Evolutionary analysis of all PCGs based on the nonsynonymous/synonymous substitution rate ratio (ω) indicated that cox1 and atp8 exhibited the lowest and highest evolutionary rates, respectively, and that the evolutionary rates of all PCGs, except for cox3, nad2, and nad3, were lower than the average ω of click beetles. Phylogenetic analyses based on concatenated and coalescent approaches indicated that Pectocera sp. was sister to Campsosternus auratus in the same tribe (Oxynopterini) with high support. This study offers insight into the mitogenomic basis of Pectocera sp. and provides an important data resource for exploring the taxonomy, phylogeny, and evolution of click beetles.

Functional Morphology of the Thorax of the Click Beetle Campsosternus auratus (Coleoptera, Elateridae), with an Emphasis on Its Jumping Mechanism.

We investigated and described the thoracic structures, jumping mechanism, and promesothoracic interlocking mechanism of the click beetle Campsosternus auratus (Drury) (Elateridae: Dendrometrinae). Two experiments were conducted to reveal the critical muscles and sclerites involved in the jumping mechanism. They showed that M2 and M4 are essential clicking-related muscles. The prosternal process, the prosternal rest of the mesoventrite, the mesoventral cavity, the base of the elytra, and the posterodorsal evagination of the pronotum are critical clicking-related sclerites. The destruction of any of these muscles and sclerites resulted in the loss of normal clicking and jumping ability. The mesonotum was identified as a highly specialized saddle-shaped biological spring that can store elastic energy and release it abruptly. During the jumping process of C. auratus, M2 contracts to establish and latch the clicking system, and M4 contracts to generate energy. The specialized thoracic biological springs (e.g., the prosternum and mesonotum) and elastic cuticles store and abruptly release the colossal energy, which explosively raises the beetle body in a few milliseconds. The specialized trigger muscle for the release of the clicking was not found; our study supports the theory that the triggering of the clicking is due to the building-up of tension (i.e., elastic energy) in the system.

The genus Cangshanaltica (Coleoptera: Chrysomelidae: Alticinae): overview, new species, and notes on species complexes.

Moss-inhabiting flea beetles form a diversified ecological group of beetles with a number of recently described new species. Here, we present an overview of known species of the moss-inhabiting flea beetle genus Cangshanaltica Konstantinov, Chamorro, Prathapan, Ge & Yang, 2013 which is distributed in Eastern and South-Eastern Asia. We describe three new species: Cangshanaltica guanxiensis sp. nov. from Guanxi, China, C. marginata sp. nov. from Yunnan, China, and C. javana sp. nov., from Java, Indonesia. Additionally, we redescribe Cangshanaltica castanea (Gruev, 1985) comb. nov., a species formerly placed in Ivalia. Based on our studies of the morphological characters, we assume that some of the currently recognized species may in fact represent species complexes. Our findings also extend the known distribution of Cangshanaltica to Indonesia. A key of all described Cangshanaltica species, and an annotated checklist are provided.

Revision of Chinese Phorocardius species (Coleoptera, Elateridae, Cardiophorinae).

The Chinese species of Phorocardius Fleutiaux, 1931 have been studied and six species are described as new: P. alterlineatus Ruan & Douglas, sp. nov.; P. flavistriolatus Ruan & Douglas, sp. nov.; P. minutus Ruan & Douglas, sp. nov.; P. rufiposterus Ruan & Douglas, sp. nov.; P. yunnanensis Ruan & Douglas, sp. nov.; and P. zhiweii Ruan, Douglas & Qiu, sp. nov. Lectotypes are designated for Cardiophorus comptus Candèze, 1860, Cardiophorus contemptus Candèze, 1860, Phorocardius magnus Fleutiaux, 1931, and Cardiophorus manuleatus Candèze, 1888. The holotype is identified for Cardiophorus yanagiharae Miwa, 1927. Phorocardius florentini (Fleutiaux, 1895) and P. manuleatus (Candèze, 1888) are newly reported from China; P. comptus (Candèze, 1860) is excluded from the Chinese fauna. A key to the 11 Phorocardius species known from China is given. Phorocardius is newly recorded from deep within the Palearctic Region. The procoxal cavities of P. rufiposterus Ruan & Douglas, sp. nov. are closed, which is different from all other species of Phorocardius. An annotated checklist of the 21 Phorocardius species of the world is provided. Additionally, Phorocardius contemptus (Candèze, 1860), comb. nov. is transferred from Cardiophorus to Phorocardius; four species are transferred from Phorocardius to Displatynychus: Displatynychus bombycinus (Candèze, 1895), comb. nov., Displatynychus pakistanicus (Platia & Ahmed, 2016), comb. nov., Displatynychus sobrinus (Laporte, 1840), comb. nov., and Displatynychus tibialis (Platia & Ahmed, 2016), comb. nov.

The Biology and Immature Stages of the Moss-Eating Flea Beetle Cangshanaltica fuanensis sp. nov. (Coleoptera, Chrysomelidae, Galerucinae, Alticini), with Description of a Fan-Driven High-Power Berlese Funnel.

The biology of the moss and leaf litter inhabiting flea beetles is poorly understood. In this study, a new species of moss-eating flea beetles Cangshanaltica fuanensis sp. nov. is described; the morphology of adult and immature stages is examined and illustrated. Its life history and biology are studied. The remarkable and unique biological features are revealed: (1) females deposit one large egg at a time, egg length equals 0.4-0.5 times the female body length, these are unusual in Chrysomelidae; (2) females have only two ovarioles on each side of the ovary, which has not been reported in other Chrysomelidae species; (3) females lay and hide each egg under a spoon-shaped moss leaf; (4) cannibalism of a second instar larva on an egg was observed. Both adults and larvae feed on moss and are polyphagous; their feces mainly consist of un-digested moss fragments; high humidity is essential for the survival of eggs and larvae and expedites the hatching. In addition, a modified fan-driven Berlese funnel is designed for faster extraction of moss inhabiting flea beetles. This device could also be used for collecting other ground-dwelling arthropods. Its working diagram is illustrated and described.

The jumping mechanism of flea beetles (Coleoptera, Chrysomelidae, Alticini), its application to bionics and preliminary design for a robotic jumping leg.

Flea beetles (Coleoptera, Chrysomelidae, Galerucinae, Alticini) are a hyperdiverse group of organisms with approximately 9900 species worldwide. In addition to walking as most insects do, nearly all the species of flea beetles have an ability to jump and this ability is commonly understood as one of the key adaptations responsible for its diversity. Our investigation of flea beetle jumping is based on high-speed filming, micro-CT scans and 3D reconstructions, and provides a mechanical description of the jump. We reveal that the flea beetle jumping mechanism is a catapult in nature and is enabled by a small structure in the hind femur called an 'elastic plate' which powers the explosive jump and protects other structures from potential injury. The explosive catapult jump of flea beetles involves a unique 'high-efficiency mechanism' and 'positive feedback mechanism'. As this catapult mechanism could inspire the design of bionic jumping limbs, we provide a preliminary design for a robotic jumping leg, which could be a resource for the bionics industry.

A review of the genus Lankaphthona Medvedev, 2001, with comments on the modified phallobase and the unique abdominal appendage of L.binotata (Baly) (Coleoptera, Chrysomelidae, Galerucinae, Alticini).

The flea beetle genus Lankaphthona Medvedev, 2001 is redescribed and a new species L.yunnantarsella sp. nov. Ruan, Konstantinov & Prathapan is described. Longitarsella Medvedev, 2009, syn. nov. and Philotarsa Medvedev, 2009, syn. nov. are newly synonymized with Lankaphthona. Philotarsalaosica Medvedev, 2009, syn. nov. is synonymized with Lankaphthonaphuketensis (Gruev, 1989). The following new combinations are proposed: Lankaphthonabinotata (Baly, 1876), comb. nov.; Lankaphthonacostata (Medvedev, 2016), comb. nov.; Lankaphthonacyanipennis (Medvedev, 2017), comb. nov.; Lankaphthonanigronotata (Jacoby, 1896), comb. nov.; Lankaphthonanotatipennis (Medvedev, 2009), comb. nov.; and Lankaphthonaphuketensis (Gruev, 1989), comb. nov., status restored. A highly specialized spoon-shaped 'appendage' is discovered on the first abdominal ventrite of males of Lankaphthonabinotata. Aedeagus of the same species has aberrant sheath-shaped phallobase encircling the median lobe. Morphology and possible function of these structures are discussed. Menispermaceae are newly reported as the host plants of the genus.

Geometric morphometric analysis of the pronotum and elytron in stag beetles: insight into its diversity and evolution.

Stag beetles (Coleoptera, Scarabaeoidea, Lucanidae) have received extensive attention from researchers in behavioral ecology and evolutionary biology. There have been no previous quantitative analyses, particularly using a geometric morphometric approach based on a large sample of data, to shed light on the morphological diversity and evolution of Lucanidae. Thoracic adaptation and ecological differentiation are intimately related, and the pronotum bears important muscles and supports the locomotion of prothoracic legs. The elytron is an autapomorphy of the Coleoptera. To reconstruct and visualize the patterns of evolutionary diversification and phylogenetic history of shape change, an ancestral groundplan can be reconstructed by mapping geometric morphometric data onto a phylogenetic tree. In this study, the morphologies of the pronotum and elytron in 1303 stag beetles (Lucanidae), including approximately 99.2% of all globally described species, were examined, thus revealing several aspects of morphological diversity and evolution. First, on the basis of geometric morphometric analysis, we found significant morphological differences in the pronotum or elytron between any two Lucanidae subfamilies. And we subsequently reconstructed the ancestral groundplans of the two structures in stag beetles and compared them with those of extant species (through cladistic and geometric morphometric methods). The ancestral groundplan of Lucanidae was found to be most similar to extant Nicagini in both the pronotum and elytron, according to Mahalanobis distances. Furthermore, we analyzed species richness and morphological diversity of stag beetles and the relationships between them and found that the two parameters were not always correlated. Aesalinae was found to be the most diverse subfamily in both the pronotum and elytron, despite its poor species richness, and the diversity of the pronotum or elytron was not superior in Lucaninae, despite its high species richness. Our study provides insights into the morphological variations and evolutionary history of the pronotum and elytron in four subfamilies of stag beetles, and it illuminates the relationship between morphological diversity and species richness. Intriguingly, our analysis indicates that morphological diversity and species richness are not always correlated. These findings may stimulate further studies in this field.

Revision of the Oriental Chaetocnema species (Coleoptera, Chrysomelidae, Galerucinae, Alticini).

The Oriental species of Chaetocnema Stephens, 1831 are revised. There are 85 valid species, including 19 new species: C. angustifrons sp. nov.; C. appendiculata sp. nov.; C. baoshanica sp. nov.; C. dapitanica sp. nov.; C. glabra sp. nov.; C. greenica sp. nov.; C. jinxiuensis sp. nov.; C. hongkongensis sp. nov.; C. latapronota sp. nov.; C. midimpunctata sp. nov.; C. nigrilata sp. nov.; C. parafusiformis sp. nov.; C. paragreenica sp. nov.; C. paraumesaoi sp. nov.; C. purerulea sp. nov.; C. reteimpunctata sp. nov.; C. sabahensis sp. nov.; C. subbasalis sp. nov.; C. trapezoida sp. nov.                Fifteen new synonyms are proposed: C. assamensis Scherer, 1969 (syn. nov.) = C. hainanensis Chen, 1932; C. birmanica Jacoby, 1892 (syn. nov.) = C. malayana Baly, 1877; C. ebenina Warchalowski, 1973 (syn. nov.) = C. yiei Kimoto, 1970; C. flavipennis Medvedev, 1996 (syn. nov.) = C. granulicollis Jacoby, 1896; C. harita Maulik, 1926 (syn. nov.) = C. westwoodi Baly, 1877; C. himalayana Medvedev, 1993 (syn. nov.) = C. melonae Chen, 1934; C. kwangsiensis Chen, 1939 (syn. nov.) = C. hainanensis Chen, 1932; C. loriae Jacoby, 1905 (syn. nov.) = C. nigrica Motschulsky, 1858; C. nepalensis Scherer, 1969 (syn. nov.) = C. bella Baly, 1877; C. nitens Baly, 1877 (syn. nov.) = C. nigrica Motschulsky, 1858; C. placida Jacoby, 1896 (syn. nov.) = C. bella Baly 1877; C. shanensis Bryant, 1939 (syn. nov.) = C. bella Baly 1877; C. subcostata Jacoby, 1889 (syn. nov.) = C. wallacei Baly, 1877; C. vietnamica Chen Wang, 1980 (syn. nov.) = C. modigliani Jacoby, 1896; C. vietnamica Medvedev, 2009 (syn. nov.) = C. wallacei Baly, 1877.                C. vietnamica Medvedev, 2001 is a new junior homonym of C. vietnamica Chen Wang, 1980.                C. melonae Chen, 1934 status restored and resurrected from synonymy with C. duvivieri Jacoby, 1892 in Medvedev, 2001: 613. Two subspecies are raised to species level: C. taiwanensis Chûjô, 1965 new status for C. tonkinensis taiwanensis Chûjô, 1965 and C. yunnanica Heikertinger, 1951 new status for C. discreta yunnanica Heikertinger, 1951.                Lectotypes are designated for 25 species: C. minuta Jacoby, 1896; C. granulicollis Jacoby, 1896; C. kwangsiensis Chen, 1939; C. longipunctata Maulik, 1926; C. montivaga Maulik, 1926; C. basalis Baly, 1877; C. parvula Baly, 1877; C. nitens Baly, 1877; C. geniculata Jacoby, 1896; C. simplicifrons (Baly, 1876); C. sticta Maulik, 1926; C. sumatrana Jacoby, 1896; C. wallacei Baly, 1877; C. alticola Maulik, 1926; C. belli Jacoby 1904; C. cognata Baly, 1877; C. squarrosa Baly, 1877; C. concinnipennis Baly, 1877; C. malayana Baly, 1877; C. birmanica Jacoby, 1892; C. merguiensis Bryant, 1941; C. pusaensis Maulik, 1926; C. singala Maulik, 1926; C. westwoodi Baly, 1877; C. harita Maulik, 1926.

Plasticity of mycangia in Xylosandrus ambrosia beetles.

Insects that depend on microbial mutualists evolved a variety of organs to transport the microsymbionts while dispersing. The ontogeny and variability of such organs is rarely studied, and the microsymbiont's effects on the animal tissue development remain unknown in most cases. Ambrosia beetles (Coleoptera: Curculionidae: Scolytinae or Platypodinae) and their mutualistic fungi are an ideal system to study the animal-fungus interactions. While the interspecific diversity of their fungus transport organ-mycangia-is well-known, their developmental plasticity has been poorly described. To determine the ontogeny of the mycangium and the influence of the symbiotic fungus on the tissue development, we dissected by hand or scanned with micro-CT the mycangia in various developmental stages in five Xylosandrus ambrosia beetle species that possess a large, mesonotal mycangium: Xylosandrus amputatus, Xylosandrus compactus, Xylosandrus crassiusculus, Xylosandrus discolor, and Xylosandrus germanus. We processed 181 beetle samples from the United States and China. All five species displayed three stages of the mycangium development: (1) young teneral adults had an empty, deflated and cryptic mycangium without fungal mass; (2) in fully mature adults during dispersal, the pro-mesonotal membrane was inflated, and most individuals developed a mycangium mostly filled with the symbiont, though size and symmetry varied; and (3) after successful establishment of their new galleries, most females discharged the bulk of the fungal inoculum and deflated the mycangium. Experimental aposymbiotic individuals demonstrated that the pronotal membrane invaginated independently of the presence of the fungus, but the fungus was required for inflation. Mycangia are more dynamic than previously thought, and their morphological changes correspond to the phases of the symbiosis. Importantly, studies of the fungal symbionts or plant pathogen transmission in ambrosia beetles need to consider which developmental stage to sample. We provide illustrations of the different stages, including microphotography of dissections and micro-CT scans.

Structure of the Ambrosia Beetle (Coleoptera: Curculionidae) Mycangia Revealed Through Micro-Computed Tomography.

Ambrosia beetles (Coleoptera: Curculionidae: Scolytinae and Platypodinae) rely on a symbiosis with fungi for their nutrition. Symbiotic fungi are preserved and transported in specialized storage structures called mycangia. Although pivotal in the symbiosis, mycangia have been notoriously difficult to study, given their minute size and membranous structure. We compared the application of novel visualization methods for the study of mycangia, namely micro-computed tomography (micro-CT) and laser ablation tomography (LATscan) with traditional paraffin sectioning. Micro-CT scanning has shown the greatest promise in new organ discovery, while sectioning remains the only method with sufficient resolution for cellular visualization. All three common types of mycangia (oral, mesonotal, and pronotal) were successfully visualized and presented for different species of ambrosia beetles: Ambrosiodmus minor (Stebbing) 1909, Euplatypus compositus (Say) 1823, Premnobius cavipennis Eichhoff 1878, Scolytoplatypus raja Blandford 1893, Xylosandrus crassiusculus (Motschulsky) 1866 and X. amputatus (Blandford) 1894. A reconstruction of the mycangium and the surrounding musculature in X. amputatus is also presented. The advantages of micro-CT compared to the previously commonly used microtome sectioning include the easy visualization and recording of three-dimensional structures, their position in reference to other internal structures, the ability to distinguish natural aberrations from technical artifacts, and the unprecedented visualizations of the anatomic context of mycangia enabled by the integrated software.

New contributions to the knowledge of Chinese flea beetle fauna (III): revision of Meishania Chen Wang with description of five new species (Coleoptera: Chrysomelidae: Galerucinae).

The flea beetle genus Meishania Chen Wang is revised and five new species-M. cangshanensis sp. nov., M. flavipennis sp. nov., M. fulvotigera sp. nov., and M. sichuanica sp. nov. from China and M. bhutanensis sp. nov. from Bhutan-are described. All species of Meishania are illustrated and a key to species is provided.

Description of two new species of Dima Charpentier, 1825 from China (Coleoptera: Elateridae: Dendrometrinae).

Dima guizhouensis Ruan, Kundrata Qiu sp. nov. and Dima xiaolongi Ruan, Kundrata Qiu sp. nov. are described from Guizhou and Hunan Provinces in China, respectively. Both species are easily distinguishable from most of their congeners by having a strongly convex pronotum and relatively long elytra with carinate interstria VI. The only superficially similar species is D. tianmuensis Qiu Kundrata in Qiu et al. (2018) from Zhejiang Province. Habitus and main diagnostic characters are figured for both new species, and their systematic placement is discussed. The discovery of these species brings the total number of Dima species in China to 13.

Contributions to the knowledge of Chinese flea beetle fauna (II): Baoshanaltica new genus and Sinosphaera new genus (Coleoptera, Chrysomelidae, Galerucinae, Alticini).

Two new genera: Baoshanalticagen. n. and Sinosphaeragen. n., and two new species: Baoshanaltica minutasp. n. and Sinosphaera apterasp. n. from south-west China are described and illustrated. Baoshanaltica is compared to the allied moss-inhabiting genera Cangshanaltica Konstantinov et al. and Phaelota Jacoby, in addition to Minota Kutschera. Sinosphaera is compared to Sphaeroderma Stephens, Omeisphaera Chen & Zia, Jacobyana Maulik, and Kamala Maulik.

Visualization of the 3D structures of small organisms via LED-SIM.

BACKGROUND: Innovative new techniques that aid in the visualization of microscopic anatomical structures have improved our understanding of organismal biology significantly. It is often challenging to observe internal 3D structures, despite the use of techniques such as confocal laser scanning microscopy (CLSM), micro-computed tomography (Micro-CT), magnetic resonance imaging (MRI), focused ion beam scanning electron microscopy (FIB-SEM) and others. In the current paper, we assess LED-SIM (DMD-based LED-illumination structured illumination microscopy), which facilitates the acquisition of nano- and micro-3D structures of small organisms in a high-resolution format (500 nm in the XY-plane and 930 nm along the Z-axis). RESULTS: We compare other microstructural imaging techniques (involving conventional optical microscopy, CLSM and Micro-CT) with LED-SIM to assess the quality (e.g. resolution, penetration depth, etc.) of LED-SIM images, as well as to document the potential short-comings of LED-SIM. Based on these results we present an optimized set of protocols to ensure that LED-SIM arthropod and nematode samples with different cuticles or textures are prepared for analysis in an optimal manner. Six arthropod and nematode specimens were tested and shown to be suitable for LED-SIM imaging, which was found to yield high resolution 3D images. CONCLUSIONS: Although LED-SIM still must be thoroughly tested before it is widely accepted and the Z-axis resolution still requires improvement, this technique offers distinct high quality 3D images. LED-SIM can be highly effective and may provide high quality 3D images for zoological studies following the guidelines of sample preparation presented in the current paper.

Penghou, a new genus of flea beetles from China (Coleoptera: Chrysomelidae: Galerucinae: Alticini).

A new genus (Penghou) with a single new species (P. yulongshan) from Yunnan Province in China is described and illustrated. It is compared to Hespera Weise, Hesperomorpha Ogloblin, Laotzeus Chen, Luperomorpha Weise, Mandarella Duvivier, Omeiana Chen, Stenoluperus Ogloblin and Taiwanohespera Kimoto.