New fossil genus and species of carrion beetle (Coleoptera,Silphidae) from the Lower Cretaceous Jinju Formation,South Korea

A new fossil genus of Silphidae, Cretosaja gen. nov., is described, based on a new fossil species, Cretosaja jinjuensis sp. nov. from the Lower Cretaceous Jinju Formation in South Korea. The association of Cretosaja with Nicrophorinae is discussed on the basis of morphological evidence. Diagnostic differences of the fossil and extant genera of Nicrophorinae are provided. Primitive characteristics presented in Mesozoic silphids are discussed.     

Characterization of cell lines developed from the colorado potato beetle, Leptinotarsa decemlineata say (coleoptera: Chrysomelidae)

In order to isolate new pathogens (viruses, microsporidia, etc.) or to evaluate the efficiency of some pathogens (serovarieties and mutants of Bacillus thuringiensis, fungi, etc.) in the control of Colorado potato beetle, an economically important pest, we established four cell lines from tissues of this insect. One was initiated from embryonated egg fragments in the M3 medium supplemented with 20% fetal bovine serum (FBS) and then transferred after several passages to the Ex-Cell 400 medium with 20% FBS. Another was initiated from larval hemocytes in Ex-Cell 400 with 5% FBS. Finally, two other cell lines were initiated from adult hemocytes: one in the Ex-Cell 400 with 20% FBS and 1% of lipid mixture and the other in the Ex-Cell 400 with 5% FBS only. These cell lines have been characterized by their morphology with light and electron microscopy, their karyotypes, cell growth, and isozyme analysis. Each cell line differed in morphologic, karyologic, growth, and isozyme patterns. The cell line initiated from embryonated eggs was growing slower than the three initiated from hemocytes. The cytotoxicity of solubilized crystal delta-endotoxins from different B. thuringiensis formulations (M-One, Trident, MYX-1806, Teknar-HPD, and Thuricide) and of destruxins, mycotoxins from Metarhizium anisopliae, was tested on these cell lines. They are sensitive to the solubilized toxins of some strains of B. thuringiensis (serovar. San Diego and serovar. tenebrionis) and to destruxins, and they can be used for the bioassay and detection of toxins and for the study of the mechanism of their action on coleopteran cells.     

The Restoration of Phytophagous Beetles in Species‐Rich Chalk Grasslands

This study focuses on the restoration of chalk grasslands over a 6‐year period and tests the efficacy of two management practices, hay spreading and soil disturbance, in promoting this process for phytophagous beetles. Restoration success for the beetles, measured as similarity to target species–rich chalk grassland, was not found to be influenced by either management practice. In contrast, restoration success for the plants did increase in response to hay spreading management. Although the presence of suitable host plants was considered to dictate the earliest point at which phytophagous beetles could successfully colonized, few beetle species colonized as soon as their host plants became established. Morphological characteristics and feeding habits of 27 phytophagous beetle species were therefore tested to identify factors that limited their colonization and persistence. The lag time between host plant establishment and colonization was greatest for flightless beetles. Beetles with foliage‐feeding larvae both colonized at slower rates than seed‐, stem‐, or root‐feeding species and persisted within the swards for shorter periods. Although the use of hay spreading may benefit plant communities during chalk grassland restoration, it did not directly benefit phytophagous beetles. Without techniques for overcoming colonization limitation for invertebrate taxa, short‐term success of restoration may be limited to the plants only.     

Genome‐wide survey of nuclear protein‐coding markers for beetle phylogenetics and their application in resolving both deep and shallow‐level divergences

Beetles (Coleoptera) are the most diverse and species‐rich insect group, representing an impressive explosive radiation in the evolutionary history of insects, and their evolutionary relationships are often difficult to resolve. The amount of ‘traditional markers’ (e.g. mitochondrial genes and nuclear rDNAs) for beetle phylogenetics is small, and these markers often lack sufficient signals in resolving relationships for such a rapidly radiating lineage. Here, based on the available genome data of beetles and other related insect species, we performed a genome‐wide survey to search nuclear protein‐coding (NPC) genes suitable for research on beetle phylogenetics. As a result, we identified 1470 candidate loci, which provided a valuable data resource to the beetle evolutionary research community for NPC marker development. We randomly chose 180 candidate loci from the database to design primers and successfully developed 95 NPC markers which can be PCR amplified from standard genomic DNA extracts. These new nuclear markers are universally applicable across Coleoptera, with an average amplification success rate of 90%. To test the phylogenetic utility, we used them to investigate the backbone phylogeny of Coleoptera (18 families sampled) and the family Coccinellidae (39 species sampled). Both phylogenies are well resolved (average bootstrap support >95%), showing that our markers can be used to address phylogenetic questions of various evolutionary depth (from species level to family level). In general, the newly developed nuclear markers are much easier to use and more phylogenetically informative than the ‘traditional markers’, and show great potential to expedite resolution of many parts in the Beetle Tree of Life.     

Sibling species of bean bruchids: a morphological and phylogenetic study of Acanthoscelides obtectus Say and Acanthoscelides obvelatus Bridwell

Acanthoscelides Schilsky is a large genus of neotropical bruchid beetles, in which most species show host plant specialization. Acanthoscelides obtectus and Acanthoscelides obvelatus are two sibling species specialized on Phaseolus beans, and are therefore considered pests. Up to now, the status of these two taxa has remained unclear, the few studies conducted having failed to elucidate whether these are two differentiated species or a single morphologically variable species. In addition, A. obvelatus has not been taken into account in the great majority of studies of bean bruchids. In this morphological and genetic study, we show that A. obtectus and A. obvelatus are two 'true' non-hybridizing species, which diverged about 22 Mya. Although the two species demonstrate only few morphological differences, we point out some diagnostic characters that enable their identification in the field. We also address a genetic method of differentiation of the two species, based on species-specific microsatellite loci. The strong morphological resemblance of these two species, despite their ancient divergence, may be the result of evolutionary stasis, which could be the consequence of stabilizing selection. Niche differentiation could enable the two species to coexist indefinitely.     

Specialists in ancient trees are more affected by climate than generalists

Ancient trees are considered one of the most important habitats for biodiversity in Europe and North America. They support exceptional numbers of specialized species, including a range of rare and endangered wood‐living insects. In this study, we use a dataset of 105 sites spanning a climatic gradient along the oak range of Norway and Sweden to investigate the importance of temperature and precipitation on beetle species richness in ancient, hollow oak trees. We expected that increased summer temperature would positively influence all wood‐living beetle species whereas precipitation would be less important with a negligible or negative impact. Surprisingly, only oak‐specialist beetles with a northern distribution increased in species richness with temperature. Few specialist beetles and no generalist beetles responded to the rise of 4°C in summer as covered by our climatic gradient. The negative effect of precipitation affected more specialist species than did temperature, whereas the generalists remained unaffected. In summary, we suggest that increased summer temperature is likely to benefit a few specialist beetles within this dead wood community, but a larger number of specialists are likely to decline due to increased precipitation. In addition, generalist species will remain unaffected. To minimize adverse impacts of climate change on this important community, long‐term management plans for ancient trees are important.     

Survival of Flower-visiting Chrysomelids during Non General-flowering Periods in Bornean Dipterocarp Forests

In SE Asian rain forests, general flowering, a community-wide synchronous flowering, occurs at irregular and supra-annual intervals. During general flowering periods (GFP), most Dipterocarpaceae and many other trees flower profusely, while flowering plants are scant between GFP. During flowerless periods, anthophilous animals that depend on floral resources for food may suffer food shortages and subsequently decrease in abundance. Flower-visiting chrysomelid adults are major pollinators for some canopy tree species that flower during GFP. Although such chrysomelids feed on flower petals, the means by which they survive flowerless periods remains unknown. We determined the abundance of flower-visiting chrysomelids in GFP and non-GFP through light trap samples and examined the effects of the presence of young leaves and flowers of dipterocarps on local abundance, and feeding preferences of flower-visiting chrysomelids. We found no clear tendency that the chrysomelid species number and the abundance during GFP were consistently higher than those during non-GFP. Chrysomelid adults were more abundant on trees with many young leaves or flowers than on trees lacking young leaves and flowers. At least a few flower-visiting chrysomelids were observed feeding on young dipterocarp leaves and visiting young leaves and flowers of non-dipterocarps in the canopy during non-GFP. All results consistently suggest that chrysomelids are able to survive flowerless periods by feeding on the young leaves of dipterocarps and on the young leaves and flowers of non-dipterocarps; through this alternate feeding, chrysomelid populations are maintained at sufficient levels to function as effective pollinators of trees that flower during GFP.     

Water balance of over-wintering beetles in relation to strategies for cold tolerance

The vapour pressure of the haemolymph of a supercooled insect is higher than the vapour pressure of the haemolymph of a frozen insect at the same temperature. The aim of the study was to see whether this may affect the water loss of freeze-avoiding and freezetolerant, over-wintering beetles. The rates of water loss were determined on freeze-tolerant Pytho depressus larvae and Upis ceramboides adults. Within each species one group was kept supercooled whereas another group was frozen. All groups were incubated at-5°C. Both species displayed significantly lower rates of water loss when they were frozen than when they were supercooled. Values of respiratory rate and water loss of freeze-avoiding and freeze-tolerant species were compared to corresponding values of desert beetles. The results indicate that the freeze-avoiding species have lower rates of cuticular water loss than freeze-tolerant species. This indicates that the freeze-avoiding species have developed more efficient water-saving mechanisms than freeze-tolerant species. The reason for this may be that the haemolymph in frozen animals will be in vapour pressure equlibrium with the ice in the hibernaculum and thus there is no danger of desiccation during winter. The supercooled insects will have a vapour pressure of the haemolymph that is higher than the vapour pressure of water in the surrounding air and will thus lose water.Abbreviations BW body weight - BW_i initial body weight - BW_t body weight at time t - P vapour pressure difference between the water in the haemolymph and the water in the air - DWL_t dry weight loss at time t - M _w rate of metabolic water production - MF_w mol fraction of water, in the haemolymph - MO₂ rate of oxygen consumption - Osm osmolality of the haemolymph - P _a vapour pressure of water in the air - P _h vapour pressure of water in the haemolymph - P _w vapour pressure of pure water - Q a constant (2,02 1 oxygen per g fat metabolized) relating oxygen consumption to dry weight loss when fat is metabolized - R a constant (1,89 1 oxygen per g water produced) relating metabolic water production to oxygen consumption when fat is metabolized - R _dwl rate of dry weight loss - RH relative humidity of the air - RWC_i initial relative water content measured by weighing - RWC_t relative water content at time t - STP standard temperature and pressure     

First discovery of fossil dung beetle brood balls and nests in the Chadian Pliocene Australopithecine levels

The Pliocene continental formations of the paleo-lake Chad system are known because of the recent discovery of the first australopithecine known west of the Rift Valley. The structures under study are found in sandstone levels associated with a rich fauna, including mammals, birds, reptiles and fishes. Analysis of the depositional environment and fauna indicates a mosaic landscape of gallery forest, savannah, grassland and ephemeral rivers interrupted by lacustrine episodes. This sandstone facies contains bioturbation in the form of sandstone balls 4-12 cm in diameter, slightly flattened at the poles. These structures are characterized by an external husk or crust and by a decimetric cavity in the upper part of the ball. Between the husk and the cavity are a number of concave laminae similar to those of a bulb, whose concavity is directed toward the upper cavity. The comparison between these structures and the brood balls of modern Scarabaeidae shows great similarity, especially in the external husk, the concave internal laminae and the chamber of the grub in the upper part of the structures. This bioturbation is interpreted as fossil brood balls of dung beetles (Coleoptera, Scarabaeidae). Fossil dung beetle brood balls are generally rare, but have been known since the 1940s. They can be very abundant in any series, as described by several authors in South America. The first fossil dung balls were described in this area in 1938 by Frenguelli and by Roselli. These authors describe elementary spherical forms of 35 mm in diameter on average, flattened at the poles and with an upper cavity. The fossil dung beetle brood balls discovered in Chad are the first in which all the internal characteristic structures are preserved. Many of them are connected by a remarkably large net of tunnels which has no parallel in the past or the present.     

Hollow oaks and beetle functional diversity: Significance of surroundings extends beyond taxonomy

Veteran hollow oaks (Quercus spp.) are keystone structures hosting high insect diversity but are declining in numbers due to intensification of land use and the abandonment of traditional management. The loss of this vital habitat is resulting in a reduction of biodiversity, and this likely has consequences for ecosystem functioning, especially if functional diversity is reduced. A considerable amount of research has been done on predictors of beetle taxonomic diversity in veteran oaks, but predictors of functional diversity have remained largely unexplored. The aim of this study was to establish whether the features and surroundings of veteran oaks are related to functional diversity within three functional groups of beetles (decomposers, predators, and flower visitors) and determine whether species richness and functional diversity within the groups are dependent on the same predictors. Sampling was carried out intermittently between 2004 and 2011 on 61 veteran oaks in Southern Norway. Of the 876 beetle species that were collected, 359 were determined to be decomposers, 284 were predators, and 85 were flower visitors. Species richness and functional diversity in all groups were consistently higher in traps mounted on veteran oaks in forests than in open landscapes. However, additional predictors differed between groups, and for species richness and functional diversity. Decomposer species richness responded to tree vitality, while functional diversity responded to habitat connectivity, predator species richness responded to regrowth of shrubs while functional diversity responded to tree circumference, and flower visitor richness and functional diversity did not respond to any additional predictors. Previous studies have found that the features and surroundings of veteran oaks are important for conservation of taxonomic diversity, and the results from this study indicate that they are also important for functional diversity within multiple functional groups.