Integrated phylogenomics and fossil data illuminate the evolution of beetles.

Beetles constitute the most biodiverse animal order with over 380 000 described species and possibly several million more yet unnamed. Recent phylogenomic studies have arrived at considerably incongruent topologies and widely varying estimates of divergence dates for major beetle clades. Here, we use a dataset of 68 single-copy nuclear protein-coding (NPC) genes sampling 129 out of the 193 recognized extant families as well as the first comprehensive set of fully justified fossil calibrations to recover a refined timescale of beetle evolution. Using phylogenetic methods that counter the effects of compositional and rate heterogeneity, we recover a topology congruent with morphological studies, which we use, combined with other recent phylogenomic studies, to propose several formal changes in the classification of Coleoptera: Scirtiformia and Scirtoidea sensu nov., Clambiformia ser. nov. and Clamboidea sensu nov., Rhinorhipiformia ser. nov., Byrrhoidea sensu nov., Dryopoidea stat. res., Nosodendriformia ser. nov. and Staphyliniformia sensu nov., and Erotyloidea stat. nov., Nitiduloidea stat. nov. and Cucujoidea sensu nov., alongside changes below the superfamily level. Our divergence time analyses recovered a late Carboniferous origin of Coleoptera, a late Palaeozoic origin of all modern beetle suborders and a Triassic-Jurassic origin of most extant families, while fundamental divergences within beetle phylogeny did not coincide with the hypothesis of a Cretaceous Terrestrial Revolution.

How to extract and analyze pollen from internal organs and exoskeletons of fossil insects?

This protocol explains how to extract pollen from fossil insects with subsequent descriptions of pollen treatment. We also describe how to document morphological and ultrastructural features with light-microscopy and electron microscopy. It enables a taxonomic assignment of pollen that can be used to interpret flower-insect interactions, foraging and feeding behavior of insects, and the paleoenvironment. The protocol is limited by the state of the fossil, the presence/absence of pollen on fossil specimens, and the availability of extant pollen for comparison. For complete details on the use and execution of this protocol, please refer to Wappler et al. (2015), Ulrich and Grímsson (2020), and Wedmann et al. (2021).

The last meal of an Eocene pollen-feeding fly.

One of the most important trophic interactions today is that between insects and their floral hosts. This biotic association is believed to have been critical to the radiation of flowering plants and many pollinating insect lineages over the last 120 million years (Ma). Trophic interactions among fossil organisms are challenging to study, and most inferences are based on indirect evidence. Fossil records providing direct evidence for pollen feeding, i.e., fossil stomach and gut contents, are exceptionally rare.1,2 Such records have the potential to provide information on aspects of animal behavior and ecology as well as plant-animal interactions that are sometimes not yet recognized for their extant relatives. The dietary preferences of short-proboscid nemestrinids are unknown, and pollinivory has not been recorded for extant Nemestrinidae.3 We analyzed the contents of the conspicuously swollen abdomen of an ca. 47.5 Ma old nemestrinid fly of the genus Hirmoneura from Messel, Germany, with photogrammetry and state-of-the-art palynological methods. The fly fed on pollen from at least four plant families-Lythraceae, Vitaceae, Sapotaceae, and Oleaceae-and presumably pollinated flowers of two extant genera, Decodon and Parthenocissus. We interpret the feeding and foraging behavior of the fly, reconstruct its preferred habitat, and conclude about its pollination role and importance in paratropical environments. This represents the first evidence that short-proboscid nemestrinid flies fed, and possibly feed to this day, on pollen, demonstrating how fossils can provide vital information on the behavior of insects and their ecological relationships with plants.

A primer of host-plant specialization in bees.

The bee-flower biological association is one of the most famous examples of insect-plant interactions, and it is axiomatic that these are of critical importance for sustaining thriving terrestrial ecosystems. Yet, the most familiar associations are often artificially managed agricultural ecosystems, reflecting an exceptionally narrow range of bee species (often only one) and a concomitantly restricted range of associated behaviors, morphologies, and mechanisms tied to pollination. Here we provide a brief account of the range of bee-floral associations encompassing floral specialization in terms of diet, behavior, and morphology. These natural associations not only promote healthy ecosystems, but also can be integrated in sustainable ways for more efficient pollination of crops by targeting bee species whose diets, behaviors, and pollen-gathering structures evolved precisely to visit such floral species rather than less efficient, and often non-native, generalists that are otherwise exploited for such purposes.

Mouthpart homologies and life habits of Mesozoic long-proboscid scorpionflies.

Mesozoic long-proboscid scorpionflies (Mesopsychoidea) provide important clues to ancient plant-pollinator interactions. Among them, the family Aneuretopsychidae is especially important because its mouthparts are vital to deciphering the early evolution of Mesopsychoidea and putatively the origin of fleas (Siphonaptera). However, the identification of mouthpart homologs among Aneuretopsychidae remains controversial because of the lack of three-dimensional anatomical data. Here, we report the first Aneuretopsychidae from Late Cretaceous Burmese amber, which have short maxillary palpi and elongate mouthpart elements consisting of one pair of galeae and one hypopharynx. Their mouthparts are identical to those of Pseudopolycentropodidae (= Dualulidae, new synonym) but are not homologous to those of Siphonaptera. Our phylogenetic analysis provides robust evidence for the debated monophyly of Mesopsychoidea. Our results suggest that the long-proboscid condition has most likely evolved once in Mesopsychoidea, independently from fleas, and further reveal the variety and complexity of mid-Cretaceous pollinating insects.

Direct evidence for eudicot pollen-feeding in a Cretaceous stinging wasp (Angiospermae; Hymenoptera, Aculeata) preserved in Burmese amber.

Angiosperms and their insect pollinators form a foundational symbiosis, evidence for which from the Cretaceous is mostly indirect, based on fossils of insect taxa that today are anthophilous, and of fossil insects and flowers that have apparent anthophilous and entomophilous specializations, respectively. We present exceptional direct evidence preserved in mid-Cretaceous Burmese amber, 100 mya, for feeding on pollen in the eudicot genus Tricolporoidites by a basal new aculeate wasp, Prosphex anthophilos, gen. et sp. nov., in the lineage that contains the ants, bees, and other stinging wasps. Plume of hundreds of pollen grains wafts from its mouth and an apparent pollen mass was detected by micro-CT in the buccal cavity: clear evidence that the wasp was foraging on the pollen. Eudicots today comprise nearly three-quarters of all angiosperm species. Prosphex feeding on Tricolporoidites supports the hypothesis that relatively small, generalized insect anthophiles were important pollinators of early angiosperms.

Beetle Pollination of Cycads in the Mesozoic.

Cycads, unlike modern wind-pollinated conifers and Ginkgo, are unusual in that they are an ancient group of gymnosperms pollinated by insects [1-3]. Although it is well documented that cycads were diverse and abundant during the mid-Mesozoic, little is known about their biogeography and pollination before the rise of angiosperms. Direct fossil evidence illuminating the evolutionary history of cycads is extremely rare [4, 5]. Here we report a specialized beetle-mediated pollination mode from the mid-Cretaceous of Myanmar, wherein a new boganiid beetle, Cretoparacucujus cycadophilus, with specialized pollen-feeding adaptations in its mouthparts and legs, was associated with many pollen grains of Cycadopites. Phylogenetic analyses indicate Cretoparacucujus as a sister group to the extant Australian Paracucujus, which pollinate the cycad Macrozamia riedlei. Our discovery, along with the current disjunct distribution of related beetle-herbivore (tribe Paracucujini) and cycad-host (tribe Encephalarteae) pairs in South Africa and Australia, indicate a probable ancient origin of beetle pollination of cycads at least in the Early Jurassic, long before angiosperm dominance and the radiation of flowering-plant pollinators later in the Cretaceous.

New fossil insect order Permopsocida elucidates major radiation and evolution of suction feeding in hemimetabolous insects (Hexapoda: Acercaria).

With nearly 100,000 species, the Acercaria (lice, plant lices, thrips, bugs) including number of economically important species is one of the most successful insect lineages. However, its phylogeny and evolution of mouthparts among other issues remain debatable. Here new methods of preparation permitted the comprehensive anatomical description of insect inclusions from mid-Cretaceous Burmese amber in astonishing detail. These "missing links" fossils, attributed to a new order Permopsocida, provide crucial evidence for reconstructing the phylogenetic relationships in the Acercaria, supporting its monophyly, and questioning the position of Psocodea as sister group of holometabolans in the most recent phylogenomic study. Permopsocida resolves as sister group of Thripida + Hemiptera and represents an evolutionary link documenting the transition from chewing to piercing mouthparts in relation to suction feeding. Identification of gut contents as angiosperm pollen documents an ecological role of Permopsocida as early pollen feeders with relatively unspecialized mouthparts. This group existed for 185 million years, but has never been diverse and was superseded by new pollenivorous pollinators during the Cretaceous co-evolution of insects and flowers. The key innovation of suction feeding with piercing mouthparts is identified as main event that triggered the huge post-Carboniferous radiation of hemipterans, and facilitated the spreading of pathogenic vectors.

Specialized and Generalized Pollen-Collection Strategies in an Ancient Bee Lineage.

Iconic examples of insect pollination have emphasized narrowly specialized pollinator mutualisms such as figs and fig wasps and yuccas and yucca moths. However, recent attention by pollination ecologists has focused on the broad spectra of pollinated plants by generalist pollinators such as bees. Bees have great impact for formulating hypotheses regarding specialization versus generalization in pollination mutualisms. We report the pollination biology of six northern European species of an extinct tribe of pollen-basket-bearing apine bees, Electrapini, of early-middle Eocene age, examined from two deposits of 48 and 44 million years in age. These bees exhibit a pattern of generalized, incidental pollen occurring randomly on their heads, thoraces, and abdomens, obtained from diverse, nectar-bearing plants. By contrast, a more restricted suite of pollen was acquired for metatibial pollen baskets (corbiculae) of the same bee taxa from a taxonomically much narrower suite of arborescent, evergreen hosts with uniform flower structure. The stereotyped plant sources of the specialist strategy of pollen collection consisted of pentamerous, radially symmetrical flowers with a conspicuous gynoecium surrounded by prominent nectar reward, organized in structurally similar compound inflorescences. Pollen specialization in bees occurs not for efficient pollination but rather in the corbiculate Electrapini as food for bee larvae (brood) and involves packing corbiculae with moistened pollen that rapidly loses viability with age. This specialist strategy was a well-developed preference by the early Eocene, providing a geochronologic midpoint assessment of bee pollen-collection strategies.

Early origin of parental care in Mesozoic carrion beetles.

The reconstruction and timing of the early stages of social evolution, such as parental care, in the fossil record is a challenge, as these behaviors often do not leave concrete traces. One of the intensely investigated examples of modern parental care are the modern burying beetles (Silphidae: Nicrophorus), a lineage that includes notable endangered species. Here we report diverse transitional silphids from the Mesozoic of China and Myanmar that provide insights into the origins of parental care. Jurassic silphids from Daohugou, sharing many defining characters of Nicrophorinae, primitively lack stridulatory files significant for parental care communications; although morphologically similar, Early Cretaceous nicrophorines from the Jehol biota possess such files, indicating that a system of parental care had evolved by this early date. More importantly, burying beetles of the genus Nicrophorus have their earliest first record in mid-Cretaceous Burmese amber, and document early evolution of elaborate biparental care and defense of small vertebrate carcasses for their larvae. Parental care in the Early Cretaceous may have originated from competition between silphids and their predators. The rise of the Cretaceous Nicrophorinae implies a biology similar to modern counterparts that typically feed on carcasses of small birds and mammals.

A complete insect from the Late Devonian period.

After terrestrialization, the diversification of arthropods and vertebrates is thought to have occurred in two distinct phases, the first between the Silurian and the Frasnian stages (Late Devonian period) (425-385 million years (Myr) ago), and the second characterized by the emergence of numerous new major taxa, during the Late Carboniferous period (after 345 Myr ago). These two diversification periods bracket the depauperate vertebrate Romer's gap (360-345 Myr ago) and arthropod gap (385-325 Myr ago), which could be due to preservational artefact. Although a recent molecular dating has given an age of 390 Myr for the Holometabola, the record of hexapods during the Early-Middle Devonian (411.5-391 Myr ago, Pragian to Givetian stages) is exceptionally sparse and based on fragmentary remains, which hinders the timing of this diversification. Indeed, although Devonian Archaeognatha are problematic, the Pragian of Scotland has given some Collembola and the incomplete insect Rhyniognatha, with its diagnostic dicondylic, metapterygotan mandibles. The oldest, definitively winged insects are from the Serpukhovian stage (latest Early Carboniferous period). Here we report the first complete Late Devonian insect, which was probably a terrestrial species. Its 'orthopteroid' mandibles are of an omnivorous type, clearly not modified for a solely carnivorous diet. This discovery narrows the 45-Myr gap in the fossil record of Hexapoda, and demonstrates further a first Devonian phase of diversification for the Hexapoda, as in vertebrates, and suggests that the Pterygota diversified before and during Romer's gap.

Diverse transitional giant fleas from the Mesozoic era of China.

Fleas are one of the major lineages of ectoparasitic insects and are now highly specialized for feeding on the blood of birds or mammals. This has isolated them among holometabolan insect orders, although they derive from the Antliophora (scorpionflies and true flies). Like most ectoparasitic lineages, their fossil record is meagre and confined to Cenozoic-era representatives of modern families, so that we lack evidence of the origins of fleas in the Mesozoic era. The origins of the first recognized Cretaceous stem-group flea, Tarwinia, remains highly controversial. Here we report fossils of the oldest definitive fleas--giant forms from the Middle Jurassic and Early Cretaceous periods of China. They exhibit many defining features of fleas but retain primitive traits such as non-jumping hindlegs. More importantly, all have stout and elongate sucking siphons for piercing the hides of their hosts, implying that these fleas may be rooted among the pollinating 'long siphonate' scorpionflies of the Mesozoic. Their special morphology suggests that their earliest hosts were hairy or feathered 'reptilians', and that they radiated to mammalian and bird hosts later in the Cenozoic.

Direct and indirect fossil records of megachilid bees from the Paleogene of Central Europe (Hymenoptera: Megachilidae).

Aside from pollen and nectar, bees of the subfamily Megachilinae are closely associated with plants as a source of materials for nest construction. Megachilines use resins, masticated leaves, trichomes and other plant materials sometimes along with mud to construct nests in cavities or in soil. Among these, the leafcutter bees (Megachile s.l.) are the most famous for their behaviour to line their brood cells with discs cut from various plants. We report on fossil records of one body fossil of a new non-leafcutting megachiline and of 12 leafcuttings from three European sites-Eckfeld and Messel, both in Germany (Eocene), and Menat, France (Paleocene). The excisions include the currently earliest record of probable Megachile activity and suggest the presence of such bees in the Paleocene European fauna. Comparison with extant leafcuttings permits the interpretation of a minimal number of species that produced these excisions. The wide range of size for the leafcuttings indirectly might suggest at least two species of Megachile for the fauna of Messel in addition to the other megachiline bee described here. The presence of several cuttings on most leaves from Eckfeld implies that the preferential foraging behaviour of extant Megachile arose early in megachiline evolution. These results demonstrate that combined investigation of body and trace fossils complement each other in understanding past biodiversity, the latter permitting the detection of taxa not otherwise directly sampled and inferences on behavioural evolution.

A giant termite from the Late Miocene of Styria, Austria (Isoptera).

A giant termite is described and figured from the Late Miocene of the Styrian Basin in southeastern Austria. Gyatermes styriensis gen. n. et sp. n. is represented by a relatively complete forewing, with basal scale. The fossil approximates in size the largest of all termites today and is the largest fossil termite on record. The presence of this species in the Late Miocene fauna of Europe indicates that climatic conditions were appropriate for the persistence of species and colonies requiring relatively stable, warm conditions. The genus is primitive in overall features but shares some similarity with the dampwood termites.