Uncovering Active Bacterial Symbionts in Three Species of Pollen-feeding Beetles (Nitidulidae: Meligethinae).

Microbial symbionts enable many phytophagous insects to specialize on plant-based diets through a range of metabolic services. Pollen comprises one-plant tissue consumed by such herbivores. While rich in lipids and proteins, its nutrient content is often imbalanced and difficult-to-access due to a digestibly recalcitrant cell wall. Pollen quality can be further degraded by harmful allelochemicals. To identify microbes that may aid in palynivory, we performed cDNA-based 16S rRNA metabarcoding on three related pollen beetles (Nitidulidae: Meligethinae) exhibiting different dietary breadths: Brassicogethes aeneus, B. matronalis, and Meligethes atratus. Nine bacterial symbionts (i.e., 97% OTUs) exhibited high metabolic activity during active feeding. Subsequent PCR surveys revealed varying prevalence of those from three Rickettsialles genera-Lariskella, Rickettsia, and Wolbachia-within beetle populations. Our findings lay the groundwork for future studies on the influence of phylogeny and diet on palynivorous insect microbiomes, and roles of symbionts in the use of challenging diets.

Underestimated diversity: Cryptic species and phylogenetic relationships in the subgenus Cobalius (Coleoptera: Hydraenidae) from marine rockpools.

Marine rockpools are isolated patches of habitat in the supratidal environment (the so-called splash zone), at the transition between sea and land, found along the rocky shores worldwide and characterized by harsh conditions for life. Nonetheless, few specialized invertebrates successfully colonized this peculiar environment. Among them several members of the water beetles Ochthebius Leach, 1815, subgenus Cobalius Rey (1886), which are found almost exclusively in supratidal and upper-most intertidal marine rockpools from the eastern Atlantic Ocean (Cape Verde, Canary Islands and Morocco, Madeira, Azores) throughout the whole Mediterranean basin. The subgenus Cobalius before 2020 was considered to include ten valid species, based on morphological differences. In late 2020, four additional new species were described. However, recent molecular phylogenetic studies have uncovered further cryptic diversity suggesting the presence of multiple undetected species within this group, highlighting that the species boundaries remain unclear and systematics and taxonomy are in need of revision. In this study we provide a molecular phylogeny based on DNA sequence data from mitochondrial and nuclear genes obtained from ten described species belonging to the subgenus Cobalius, and extensive taxon sampling, in order to better understand the phylogenetic relationships within this genus and to infer the biogeographic processes behind its diversification. We also used a molecular clock to define a time window for diversification of distinct clades within the subgenus, and explore aspects of its evolutionary history. Finally, we used three species delimitation methods (PTP, GMYC and ABGD) to clarify taxonomy and validate species boundaries. Our phylogenetic and biogeographic results identified sixteen independent lineages grouped in four main clades and the possible origin of Cobalius was estimated to be in the Early Miocene (~22 Mya) in W Mediterranean area. Moreover, species delimitation methods suggest there are between 16 and 24 putative species, most of them diverged during the Late Miocene, Pliocene and Pleistocene (6.0-0.11 Mya).

Rosaceae, Brassicaceae and pollen beetles: exploring relationships and evolution in an anthophilous beetle lineage (Nitidulidae, Meligethes-complex of genera) using an integrative approach.

BACKGROUND: Meligethes are pollen-beetles associated with flowers of Rosaceae as larvae. This genus currently consists of 63 known species in two subgenera, Meligethes and Odonthogethes, predominantly occurring in the eastern Palaearctic. We analyzed 74 morphological and ecological characters (169 states) of all species, as well as of 11 outgroup species from 7 Meligethinae genera (including Brassicogethes), to investigate their phylogeny. We also conducted a parallel molecular analysis on 9 Meligethes, 9 Odonthogethes, 3 Brassicogethes and 2 Meligethinus species based on DNA sequence data from mitochondrial (COI, 16S) and nuclear (CAD) genes. RESULTS: Morphological phylogenetic reconstructions supported the monophyly of the whole genus and clades corresponding to purported subgenera Meligethes s.str. and Odonthogethes. Main species-groups were mostly confirmed, however some unresolved polytomies remained. Molecular data placed members of Brassicogethes (including 42 mostly W Palearctic species associated with Brassicaceae) as sister to Odonthogethes, with this clade being sister to Meligethes s.str. This phylogenetic scenario suggests that monophyletic Meligethes s.str., Odonthogethes and Brassicogethes should be regarded alternatively as three subgenera of a monophyletic Meligethes, or three genera in a monophyletic genus-complex, with mutually monophyletic Brassicogethes and Odonthogethes. Molecular analyses estimated the origin of this lineage at ca. 14-15 Mya from a common stem including Meligethinus. CONCLUSIONS: We hypothesize that the ancestor of Meligethes specialized on Rosaceae in the Middle Miocene (likely in Langhian Age) and subsequently radiated during Late Miocene and Plio-Pleistocene maintaining a trophic niche on this plant family. This radiation was primarily due to geographic isolation in E Asiatic mountain systems. Combined evidence from morphology, ancestral state parsimony reconstruction of host-plant associations and molecular evidence suggested that Rosoideae (Rosa spp.) represented the ancestral hosts of Meligethes s.str., followed by an independent shift of ancestral Odonthogethes (ca. 9-15 Mya) on Rubus (Rosoideae) and members of Rosaceae Spiraeoideae. Other ancestral Odonthogethes probably shifted again on the unrelated plant family Brassicaceae (maybe 8-14 Mya in S China), allowing a rapid westward radiation of the Brassicogethes clade.

Integrative taxonomy of Teucriogethes pollen beetles (Coleoptera: Nitidulidae: Meligethinae), with implications on the systematics of the genus Teucrium (Lamiaceae).

In highly specialised phytophagous insects, important insights on phylogeny of the involved insect group can be often inferred from the evolutionary history of their host plants, and vice-versa, as in the case for a new, peculiar species of pollen beetles, recently discovered in Central China (Hubei Province). This new species is described herein as Teucriogethes huangae Liu & Audisio, sp. nov., and information on its ecology and systematic position is provided. The new species, associated as larvae with a strongly isolated member of the genus Teucrium L. (Lamiaceae), T. ornatum Hemsl., exhibiting a problematic taxonomic position, represents in turn an isolated and morphologically rather aberrant taxon in its lineage, probably the largest in size (2.2-2.8mm), and the first endemic representative of its genus in the whole Eastern Palaearctic. An updating of the taxonomic assessment of members of Teucriogethes and on possible evolutionary relationships with their larval host plants, based on an integrative taxonomy approach, is finally presented. A key to identification of all known species is also introduced. ZooBank: urn:lsid:zoobank.org:pub:E349044B-29A5-416C-85CA-56143C7A29BE.

The First Two Complete Mitochondrial Genomes for the Subfamily Meligethinae (Coleoptera: Nitidulidae) and Implications for the Higher Phylogeny of Nitidulidae.

The phylogenetic status of the family Nitidulidae and its sister group relationship remain controversial. Also, the status of the subfamily Meligethinae is not fully understood, and previous studies have been mainly based on morphology, molecular fragments, and biological habits, rather than the analysis of the complete mitochondrial genome. Up to now, there has been no complete mitochondrial genome report of Meligethinae. In this study, the complete mitochondrial genomes of Meligethinus tschungseni and Brassicogethes affinis (both from China) were provided, and they were compared with the existing complete mitochondrial genomes of Nitidulidae. The phylogenetic analysis among 20 species of Coleoptera was reconstructed via PhyloBayes analysis and Maximum likelihood (ML) analysis, respectively. The results showed that the full lengths of Meligethinus tschungseni and Brassicogethes affinis were 15,783 bp and 16,622 bp, and the AT contents were 77% and 76.7%, respectively. Each complete mitochondrial genome contains 13 protein-coding genes (PCGs), 22 transfer RNA genes (tRNAs), 2 ribosomal RNA genes (rRNAs), and a control region (A + T-rich region). All the PCGs begin with the standard start codon ATN (ATA, ATT, ATG, ATC). All the PCGs terminate with a complete terminal codon, TAA or TAG, except cox1, cox2, nad4, and nad5, which terminate with a single T. Furthermore, all the tRNAs have a typical clover-leaf secondary structure except trnS1, whose DHU arm is missing in both species. The two newly sequenced species have different numbers and lengths of tandem repeat regions in their control regions. Based on the genetic distance and Ka/Ks analysis, nad6 showed a higher variability and faster evolutionary rate. Based on the available complete mitochondrial genomes, the results showed that the four subfamilies (Nitidulinae, Meligethinae, Carpophilinae, Epuraeinae) of Nitidulidae formed a monophyletic group and further supported the sister group relationship of Nitidulidae + Kateretidae. In addition, the taxonomic status of Meligethinae and the sister group relationship between Meligethinae and Nitidulinae (the latter as currently circumscribed) were also preliminarily explored.

A new Chinese Cyclogethes pollen beetle, with an updated key to species of the genus and notes on its phylogenetic positioning (Coleoptera: Nitidulidae: Meligethinae).

A peculiar new species of the genus Cyclogethes Kirejtshuk, 1979, C. tibialis sp. nov., is described from Southwestern China (Yunnan). The new species appears to be morphologically rather isolated from the other known members of this essentially Oriental genus (including half a dozen species from Northern Indian subcontinent, Northern Indochina, and Southwestern China). However, it could be more closely related to C. abnormis Kirejtshuk, 1979 from Northern India, Indochina, and Southwestern China, and to C. aldridgei Kirejtshuk, 1980 from Northern India and Nepal, from which it is easily distinguished by the more elongate body shape, and by the markedly sinuate hind tibiae in both sexes (a very unusual character state in Meligethinae, where only males of some species exhibit sexual secondary characters in the tibial shape). The new species also differs from other known taxa of the genus by the shape of the male and female genitalia. The larval hostplants of members of Cyclogethes are thus far unknown, although some clues, also involving the new species described herein, may suggest a relationship with small trees or shrubs of the family Asteraceae. Preliminary and incomplete molecular data on a studied member of the genus (C. abnormis) seems to not disagree with a phylogenetic positioning of Cyclogethes in a clade including the African genera Tarchonanthogethes Audisio & Cline, 2009, its allied Afrotropical taxa, and the Palaearctic genera Meligethes Stephens, 1830 and Brassicogethes Audisio & Cline, 2009. The article includes an updated identification key for all six known species of this genus and an updated map of their known geographic distribution.

Genome-wide diversity, population structure and signatures of inbreeding in the African buffalo in Mozambique.

The African buffalo, Syncerus caffer, is a key species in African ecosystems. Like other large herbivores, it plays a fundamental role in its habitat acting as an ecosystem engineer. Over the last few centuries, African buffalo populations have declined because of range contraction and demographic decline caused by direct or indirect human activities. In Mozambique, historically home to large buffalo herds, the combined effect of colonialism and subsequent civil wars has created a critical situation that urgently needs to be addressed. In this study, we focused on the analysis of genetic diversity of Syncerus caffer caffer populations from six areas of Mozambique. Using genome-wide SNPs obtained from ddRAD sequencing, we examined the population structure across the country, estimated gene flow between areas under conservation management, including national reserves, and assessed the inbreeding coefficients. Our results indicate that all studied populations of Syncerus caffer caffer are genetically depauperate, with a high level of inbreeding. Moreover, buffaloes in Mozambique present a significant population differentiation between southern and central areas. We found an unexpected genotype in the Gorongosa National Park, where buffaloes experienced a dramatic population size reduction, that shares a common ancestry with southern populations of Catuane and Namaacha. This could suggest the past occurrence of a connection between southern and central Mozambique and that the observed population structuring could reflect recent events of anthropogenic origin. All the populations analysed showed high levels of homozygosity, likely due to extensive inbreeding over the last few decades, which could have increased the frequency of recessive deleterious alleles. Improving the resilience of Syncerus caffer caffer in Mozambique is essential for preserving the ecosystem integrity. The most viable approach appears to be facilitating translocations and re-establishing connectivity between isolated herds. However, our results also highlight the importance of assessing intraspecific genetic diversity when considering interventions aimed at enhancing population viability such as selecting suitable source populations.

Description of the larva of Ochthebius capicola (Coleoptera: Hydraenidae) from marine rockpools of South Africa.

Larval instars II and III of Ochthebius (Ochthebius) capicola (Pdringuey 1892) from South Africa are described and illustrated by SEM, including a detailed analysis of their chaetotaxy and porotaxy. Specimens used in this study were collected with adults of the same species in marine rockpools. The larval morphology of O. capicola is compared to that of other members of the same genus occurring in this habitat, contributing to the current taxonomic and phylogenetic debate on the intrageneric taxonomy of Ochthebiinae.

Morphological Study of the Alimentary Canal and Malpighian Tubules in the Adult of the Pollen Beetle Meligethes (Odonthogethes) chinensis (Coleoptera: Nitidulidae: Meligethinae).

Meligethes (Odonthogethes) chinensis is a highly specialized species of Nitidulidae in China that takes pollen as its main food source, and its main host plant is Rubus idaeus L. (Rosaceae). In this study, the structural morphology of the alimentary canal and Malpighian tubules of adult M. (O.) chinensis was observed under light, fluorescence, and scanning electron microscopy. The alimentary canal of adult M. (O.) chinensis is divided into foregut, midgut, and hindgut. The foregut is the shortest and consists of the pharynx, esophagus, proventriculus, and cardiac valve. The midgut is a straight, distended, cylindrical, thin-walled tube. Numerous blunt-fingered gastric ceca are distributed irregularly throughout the midgut. The hindgut is subdivided into the ileum, colon, and rectum. The ileum is coiled. The colon gradually enlarges posteriorly. The rectum is thickly muscled and followed by a membranous structure. The openings of proximal Malpighian tubules are evenly inserted into the junction of the midgut and hindgut, and distal Malpighian tubules are evenly attached to the colon to form a cryptonephridial system. In this study, we also compare the structure and infer the function of the alimentary canal and Malpighian tubules among beetles, as well as discuss the evolutionary and taxonomical implications.

Scanning Electron Microscope Study of Antennae and Mouthparts in the Pollen-Beetle Meligethes (Odonthogethes) chinensis (Coleoptera: Nitidulidae: Meligethinae).

Meligethes (Odonthogethes) chinensis is a common Chinese phytophagous species in the family Nitidulidae. Its main larval host plant is Rubus idaeus L. (Rosaceae), and adults feed on pollen and other floral parts. In this study, we used scanning electron microscopy (SEM) to observe and study the fine morphology of sensilla on the antennae and mouthparts of M. chinensis. The results show that there are six types and twelve subtypes of sensilla on male antennae; seven types and fourteen subtypes on female antennae; seven types and seventeen subtypes on male mouthparts; seven types and sixteen subtypes on female mouthparts. Sensilla coeloconica (SCo) are found on the female antennae of M. chinensis only, and they are also reported on the antennae of Nitidulidae for the first time. SCo2 on the labrum present sexual dimorphism, and one subtype of sensilla basiconica (SB6) is presented on the tip of maxillary and labial palps of the male only, while other types of sensilla are very similar on the mouthparts of male and female. Finally, by comparing similar sensilla in other insects, we also attempted to discuss the functions of all sensilla on the antennae and mouthparts of M. chinensis.

Comparative Mitogenomic Analysis of Two Longhorn Beetles (Coleoptera: Cerambycidae: Lamiinae) with Preliminary Investigation into Phylogenetic Relationships of Tribes of Lamiinae.

The subfamily Lamiinae is the most taxonomically diverse subfamily of Cerambycidae, but relationships between tribes of Lamiinae are still unresolved. In order to study the characteristics of the mitogenomes of Lamiinae and the tribal-level phylogenetic relationships, we sequenced the mitogenomes of two species representing two tribes, Agapanthia amurensis (Agapanthiini) and Moechotypa diphysis (Ceroplesini), with a total length of 15,512 bp and 15,493 bp, respectively. The gene arrangements of these two new mitogenomes were consistent with the inferred ancestral insect mitogenomes. Each species contained 37 typical mitochondrial genes and a control region (A + T-rich region), including 13 protein-coding genes (PCGs), 22 transfer RNA genes (tRNAs), and two ribosomal RNA genes (rRNAs). All PCGs initiated with the standard start codon ATN, and terminated with the complete stop codons of TAA and TAG, or incomplete stop codon T. All tRNAs could be folded into a clover-leaf secondary structure except for trnS1, in which the dihydrouridine (DHU) arm was reduced. Moreover, we studied the phylogenetic relationships between some tribes of Lamiinae based in mitochondrial PCGs in nucleotides; our results show that the relationships were as follows: (Onciderini + ((Apomecynini + Acanthocinini) + ((Ceroplesini + Agapanthiini) + ((Mesosini + Pteropliini) + ((Dorcaschematini + (Saperdini 1 + (Phytoeciini + Saperdini 2))) + (Batocerini + Lamiini)))))).

Five new species of Lamiogethes Audisio amp; Cline from China (Coleoptera: Nitidulidae: Meligethinae).

Five new species of the genus Lamiogethes Audisio Cline are described from western, central and southern China: L. falcatus sp. n. (Sichuan), L. hastipenis sp. n. (Hubei), L. sagittalis sp. n. (Shaanxi), L. unditibiis sp. n. (Chongqing), and L. limaelytralis sp. n. (Sichuan). The first four new species appear to be morphologically related to Lamiogethes ancestor (Kirejtshuk, 1980) (comb. n.) from Central China, and allied taxa. Lamiogethes limaelytralis sp. n. appears to be more isolated, probably with relationships to the recently described L. forcipenis Liu, Huang, Cline Audisio, 2017 and L. convexistrigosus Liu, Huang, Cline Audisio, 2017 from Sichuan. All new species differ from other closely related taxa by details of the body shape and color, as well as the shape of the male and/or female genitalia.

Palms and pollen beetles: two new anthophilous beetle species of Meligethinus from Mozambique (Coleoptera: Nitidulidae: Meligethinae).

The genus Meligethinus Grouvelle, 1906 represents a small group of pollen beetles, including some twenty species distributed from southern Palaearctic areas to northern Oriental and Afrotropical regions. All constituent species appear to be strictly associated as larvae and adults to male inflorescences of palms (Monocots: Arecaceae). Two new species of this genus (Meligethinus mondlanei sp. nov. and M. hamerlae sp. nov.) were recently discovered in southern Mozambique, and are described herein. Both are associated as larvae with male inflorescences of the widespread eastern African palm Phoenix reclinata Jacq. The two new species are compared with related taxa from central Africa. Additionally, the presence of additional species of the same genus in southern Mozambique is reported, and the local associations of several Meligethinus species are discussed.

New species and records of pollen and sap beetles for Iran (Coleoptera: Kateretidae, Nitidulidae).

During recent expeditions in North and Southwest Iran, 10 species of Kateretidae and Nitidulidae (Coleoptera) were found. One species of Kateretidae, Brachyleptus bicoloratus Reitter, 1896, and three nitidulid species, Afrogethes schilskyi (Reitter, 1897), Stachygethes khnzoriani (Kirejtshuk, 1979), and S. nigerrimus (Rosenhauer, 1856) are recorded for the first time for the Iranian fauna (the latter is also a new record for Asia). Thymogethes ahriman (Jelínek, 1981) is herein resurrected to species rank, and two species, Thymogethes kassites sp. nov. and T. khorasanicus sp. nov., are described as new. An updated key to the known Near East and Afghan species of the genus Thymogethes Audisio & Cline, 2009 is also provided. Available and recently collected biological and distributional data, as well as short taxonomic comments, are given for the discussed species.

Revision of the African pollen beetle genera Tarchonanthogethes and Xenostrongylogethes, with insect-host plant relationships, identification key, and cladistic analysis of the Anthystrix genus-complex (Coleoptera: Nitidulidae: Meligethinae).

The Afrotropical endemic pollen beetle genera Tarchonanthogethes Audisio & Cline and Xenostrongylogethes Audisio & Cline, of the Anthystrix genus-complex, are revised. Eleven new species of Tarchonanthogethes (T. autumnalis, sp. nov., T. bisignatus, sp. nov., T. fasciatus, sp. nov., T. gratiellae, sp. nov., T. hermani, sp. nov., T. hystrix, sp. nov., T. lilliputianus, sp. nov., T. maasai, sp. nov., T. manconiae, sp. nov., T. pectinipes, sp. nov., T. thalycriformis, sp. nov.) and one new Xenostrongylogethes (X. cychramoides, sp. nov.) are described, illustrated and compared with related taxa. Tarchonanthogethes hirtus Kirejtshuk & Easton, 1988 is synonymized with T. martini (syn. nov.). Meligethes assutus Easton, 1960 from Kenya is transferred from Afrogethes Audisio & Cline to Tarchonanthogethes (comb. nov.). Meligethes singularis Grouvelle, 1919 from southern Africa is transferred from Tarchonanthogethes to Meligethinus Grouvelle, 1906 (comb. nov.). Larval host-plants for Tarchonanthogethes and Xenostrongylogethes include dioecious bushes and trees of Tarchonantheae Asteraceae (genera Brachylaena R.Br. and Tarchonanthus L.). All species currently attributed to the genera Anthystrix Kirejtshuk, Sebastiangethes Audisio, Kirk-Spriggs & Cline, Tarchonanthogethes and Xenostrongylogethes (Anthystrix genus-complex) are included in a morphology-based cladistic analysis to provide a rigorous hypothesis of phylogenetic relationships. An identification key to all 25 known species in the Anthystrix genus-complex, including all available data on insect host plant relationships, is presented.

Fauna Europaea: Coleoptera 2 (excl. series Elateriformia, Scarabaeiformia, Staphyliniformia and superfamily Curculionoidea).

Fauna Europaea provides a public web-service with an index of scientific names (including synonyms) of all living European land and freshwater animals, their geographical distribution at country level (up to the Urals, excluding the Caucasus region), and some additional information. The Fauna Europaea project covers about 230,000 taxonomic names, including 130,000 accepted species and 14,000 accepted subspecies, which is much more than the originally projected number of 100,000 species. This represents a huge effort by more than 400 contributing specialists throughout Europe and is a unique (standard) reference suitable for many users in science, government, industry, nature conservation and education. Coleoptera represent a huge assemblage of holometabolous insects, including as a whole more than 200 recognized families and some 400,000 described species worldwide. Basic information is summarized on their biology, ecology, economic relevance, and estimated number of undescribed species worldwide. Little less than 30,000 species are listed from Europe. The Coleoptera 2 section of the Fauna Europaea database (Archostemata, Myxophaga, Adephaga and Polyphaga excl. the series Elateriformia, Scarabaeiformia, Staphyliniformia and the superfamily Curculionoidea) encompasses 80 families (according to the previously accepted family-level systematic framework) and approximately 13,000 species. Tabulations included a complete list of the families dealt with, the number of species in each, the names of all involved specialists, and, when possible, an estimate of the gaps in terms of total number of species at an European level. A list of some recent useful references is appended. Most families included in the Coleoptera 2 Section have been updated in the most recent release of the Fauna Europaea index, or are ready to be updated as soon as the FaEu data management environment completes its migration from Zoological Museum Amsterdam to Berlin Museum für Naturkunde.