Uncovering Norway: Descriptions of Four New Aphidiinae Species (Hymenoptera, Braconidae) with Identification Key and Notes on Phylogenetic Relationships of the Subgenus Fovephedrus Chen.

With only 33 reported species, Norway ranks among the European countries with the lowest documented diversity of parasitoids from the subfamily Aphidiinae. The "MUST Malaise" project, carried out by Museum Stavanger in Norway, aimed to assess insect abundance and biodiversity and create a reference base for future studies. The preliminary results of our study revealed four species new to science, indicating that the current number of recorded species in Norway is significantly lower than the actual diversity. All species possess unique combinations of morphological characters, distinguishing them from other known Aphidiinae species. Molecular analysis of the barcoding region confirmed that these specimens all belong to the previously undescribed species. In this study, we describe Aphidius norvegicus sp.n., Praon breviantennalis sp.n., Ephedrus gardenforsi sp.n., and Ephedrus borealis sp.n., all collected in Norway. We also provide an identification key and discuss the phylogenetic relationships within the subgenus Fovephedrus Chen, 1986.

It's time for Africa - hidden diversity of the Aphidius colemani species group (Hymenoptera, Braconidae, Aphidiinae) south of the Sahara.

Aphidius colemani is an important biological control agent, used in greenhouses and open fields against aphid pests. Despite this economical importance, A. colemani, along with A. transcaspicus and A. platensis, has gone through a complex taxonomical history. The three species have only recently gained status as separate species again, comprising the morphologically defined Aphidius colemani species group. Other than sporadic records probably as a consequence of escape from greenhouses, the A. colemani species group members prefer warmer regions and there are numerous records from South America, Southern Europe and Asia. Based on slide-mounted material collected in the period 1964-2001 in Africa, we describe five new species belonging to this group, and report A. colemani, A. transcaspicus and A. platensis from several African countries. This data opens questions about the origin of the group and presents potential for the diversification of biological control agents against aphid pests.

Four New Species of Pseudephedrus (Hymenoptera, Braconidae, Aphidiinae) from Chile with a Key to Species Identification.

The following work consists of the description of four new species of the genus Pseudephedrus Starý (Aphidiinae), endemic to South America, associated with endemic callaphidid aphid hosts. The descriptions of the new species are based on new samples from Chile and Argentina. The new species described here are as follows: Pseudephedrus staryi sp.n., which is morphologically very close to the already described P. chilensis Starý, with clear apomorphies; P. pubescens sp.n., which presents a setose scutellum, P. flava sp.n. and P. patagonicus sp.n. which have distinct, very elongated, and crenulated petioles representing synapomorphies and probably form separate phylogenetic lineages within Pseudephedrus. All hosts are aphids from the genus Neuquenaphis Blanchard, with varying degrees of diet specialization. From our field and laboratory observations, we hypothesize that, since attempts to sample parasitoids using sweep nets were much more successful than sampling from collected aphids, and since we found mummified aphids only on the ground among the fallen leaves under the trees, Neuquenaphis aphids fall to the ground showing a dropping behaviour as a defence against natural enemies after being stung by parasitoids. This makes rearing from live aphids very unsuccessful and could help explain why it has been difficult to collect and describe species. We supplement the distribution of Pseudephedrus from South America and present a key for the identification of all species based on their morphology.

Dyscrituluseuropaeus sp. nov. (Hymenoptera, Braconidae, Aphidiinae): description of a new aphid parasitoid species with an identification key for species of the genus.

The braconid genus Dyscritulus Hincks is a small member of the subfamily Aphidiinae, distributed in Europe and Central Asia. All its species are highly specialized parasitoids of aphids of the genera Drepanosiphum Koch and, probably, Periphyllus van der Hoeven which are mostly associated with maple and sycamore trees (genus Acer). Upon examination of specimens from the Naturalis Biodiversity Center, Leiden, we unexpectedly noted unusual variability in morphological characters compared to other known Dyscritulus species. Further inspection of other material previously identified as Dyscritulusplaniceps Marshall, 1896 revealed additional specimens with the same morphological variability. Here we describe a new species of the genus, Dyscrituluseuropaeussp. nov., associated with Drepanosiphum aphids on Acer.

Two new species of subtribe Monoctonina Mackauer from Europe (Hymenoptera, Braconidae, Aphidiinae).

Falciconus hispanicus sp. n. and Monoctonus montenegrinus sp. n. are described as new members of the subtribe Monoctonina from Spain and from high mountain habitats of Montenegro, respectively. The main diagnostic characters of the newly described species and their host plant-host aphid associations are discussed.

Cereal Aphid Parasitoids in Europe (Hymenoptera: Braconidae: Aphidiinae): Taxonomy, Biodiversity, and Ecology.

Cereals are very common and widespread crops in Europe. Aphids are a diverse group of herbivorous pests on cereals and one of the most important limiting factors of cereal production. Here, we present an overview of knowledge about the taxonomy, biodiversity, and ecology of cereal aphid parasitoids in Europe, an important group of natural enemies contributing to cereal aphid control. We review the knowledge obtained from the integrative taxonomy of 26 cereal aphid primary parasitoid species, including two allochthonous species (Lysiphlebus testaceipes and Trioxys sunnysidensis) and two recently described species (Lipolexis labialis and Paralipsis brachycaudi). We further review 28 hyperparasitoid species belonging to three hymenopteran superfamilies and four families (Ceraphronoidea: Megaspillidae; Chalcidoidea: Pteromalidae, Encyrtidae; Cynipoidea: Figitidae). We also compile knowledge on the presence of secondary endosymbionts in cereal aphids, as these are expected to influence the community composition and biocontrol efficiency of cereal aphid parasitoids. To study aphid-parasitoid-hyperparasitoid food webs more effectively, we present two kinds of DNA-based approach: (i) diagnostic PCR (mainly multiplex PCR), and (ii) DNA sequence-based methods. Finally, we also review the effects of landscape complexity on the different trophic levels in the food webs of cereal aphids and their associated parasitoids, as well as the impacts of agricultural practices and environmental variation.

Sizing the Knowledge Gap in Taxonomy: The Last Dozen Years of Aphidiinae Research.

Taxonomic impediment is one of the main roadblocks to managing the current biodiversity crisis. Insect taxonomy is the biggest contributor to the taxonomic impediment, both in terms of the knowledge gap and the lack of experts. With this study, we tried to size the knowledge gap by analyzing taxonomical studies on the subfamily Aphidiinae (Hymenoptera: Braconidae) conducted from 2010 to 2021. All available taxonomic knowledge gathered in this period is critically summarized: newly described species, detection of alien species, published identification keys, etc. All findings are discussed relative to the current state of general taxonomy. Future prospects for taxonomy are also discussed.

Resolving the Taxonomic Status of Potential Biocontrol Agents Belonging to the Neglected Genus Lipolexis Förster (Hymenoptera, Braconidae, Aphidiinae) with Descriptions of Six New Species.

Lipolexis is a small genus in the subfamily Aphidiinae represented by one species in Europe (Lipolexis gracilis Förster) and by four in Asia (Lipolexis wuyiensis Chen, L. oregmae Gahan, L. myzakkaiae Pramanik and Raychaudhuri and L. pseudoscutellaris Pramanik and Raychaudhuri). Although L. oregmae is employed in biological control programs against pest aphids, the last morphological study on the genus was completed over 50 years ago. This study employs an integrative approach (morphology and molecular analysis (COI barcode region)), to examine Lipolexis specimens that were sampled worldwide, including specimens from BOLD database. These results establish that two currently recognized species of Lipolexis (L. gracilis, L. oregmae) are actually a species complex and also reveal phylogenetic relationships within the genus. Six new species are described and a global key for the identification of Lipolexis species is provided.

Phylogeny of the Subtribe Monoctonina (Hymenoptera, Braconidae, Aphidiinae).

Members of the Monoctonina subtribe have long been neglected in applied studies of the subfamily Aphidiinae, due to their low economic importance, as they do not parasitize pests of cultivated plants. Consequently, data about this group are scarce, including its taxonomy and phylogeny. In the present study, we explore inter- and intraspecific genetic variation of Monoctonina species, including genera Monoctonus Haliday 1833, Monoctonia Starý 1962, Falciconus Mackauer 1959 and Harkeria Cameron 1900. We employ two molecular markers, the barcode region of the mitochondrial cytochrome c oxidase subunit I (COI) and the D2 region of the 28S nuclear gene (28S rDNA), to analyze genetic structuring and phylogeny of all available Monoctonina species, and combine them with morphological data for an integrative approach. We report one new species, and three potentially new species which can be formally described when further specimens are available. Analysis of phylogenetic relationships within the subtribe shows a basal position for the genera Falciconus and Monoctonia, and the close relatedness of Harkeria and Monoctonus.

Review of the world Monoctonina Mackauer 1961 (Hymenoptera, Braconidae, Aphidiinae): key for their identification and descriptions of five new species.

The subtribe Monoctonina has long been overlooked when it comes to morphological studies, probably due to the low economic importance of its members. Mainly distributed in high montane areas and forest ecosystems, species of Monoctonina are not often found in large populations and do not parasitize pests of cultivated plants. Our research uncovered five new species belonging to this subtribe (Monoctonus brachyradius sp. n., M. canadensis sp. n., M. inexpectatus sp. n., M. luteus sp. n., and M. parvipalpus sp. n.). We also redescribe species originally described over a century ago, and discuss the utility of certain morphological characters, as well as status of some species within Monoctonina.

Phylogenetic relationships and subgeneric classification of European Ephedrus species (Hymenoptera, Braconidae, Aphidiinae).

In this study two molecular markers were used to establish taxonomic status and phylogenetic relationships of Ephedrus subgenera and species distributed in Europe. Fifteen of the nineteen currently known species have been analysed, representing three subgenera: Breviephedrus Gärdenfors, 1986, Lysephedrus Starý, 1958 and Ephedrus Haliday, 1833. The results of analysis of COI and EF1α molecular markers and morphological studies did not support this classification. Three clades separated by the highest genetic distances reported for the subfamily Aphidiinae on intrageneric level. Ephedrus brevis is separated from persicae and plagiator species groups with genetic distances of 19.6 % and 16.3 % respectively, while the distance between persicae and plagiator groups was 20.7 %. These results lead to the conclusion that the traditional subgeneric classification of Ephedrus needs revision. Species from persicae species group are raised to subgenus level as Fovephedrus Chen, 1986 and Lysephedrus syn. nov. is assigned as a junior synonym of subgenus Ephedrus. Key for identification of Ephedrus subgenera is provided. Ephedrus hyadaphidis Kocic & Tomanovic sp. nov. is described and several species are confirmed as valid species for the first time. Furthermore, two species are synonymised: E. dysaphidis syn. nov. as a junior synonym of E. cerasicola and E. blattnyi syn. nov. as a junior synonym of E. plagiator.

Integrative taxonomy of root aphid parasitoids from the genus Paralipsis (Hymenoptera, Braconidae, Aphidiinae) with description of new species.

Species from the genus Paralipsis are obligatory endoparasitoids of root aphids in the Palaearctic. It is known that these species are broadly distributed, parasitizing various aphid hosts and showing great biological and ecological diversity. On the other hand, this group of endoparasitoids is understudied and was thought to be represented by a single species in Europe, viz., Paralipsisenervis (Nees). However, recent description of two new species indicated the possibility of cryptic speciation and recognition of additional Paralipsis species in Europe. In this research, Paralipsis specimens collected during the last 60 years from eight European countries, as well as one sample from Morocco, were subjected to molecular and morphological characterization. Newly designed genus-specific degenerative primers successfully targeted short overlapping fragments of COI of the mitochondrial DNA. Molecular analyses showed clear separation of four independent lineages, two of which are the known species P.enervis and P.tibiator, while two new species are described here, viz., P.brachycaudi Tomanovic & Starý, sp. n. and P.rugosa Tomanovic & Starý, sp. n. No clear specialization of the taxa to a strict root aphid host has been determined. The recognized mitochondrial lineages were distinct one from another, but with a substantial within-lineage divergence rate, clearly indicating the complexity of this group of parasitoids, on which further research is required in order to clarify the factors triggering their genetic differentiation. We reviewed literature data and new records of Paralipsisenervis aphid host associations and distributions. A key for the identification of all known Paralipsis species is provided and illustrated.

Key for identification of the parasitoids (Hymenoptera: Braconidae: Aphidiinae) of aphids infesting alfalfa in Europe.

Although Aphidiinae parasitoids have been used as agents for biocontrol of alfalfa aphids for more than half a century and have been the subject of numerous ecological investigations, there is an evident lack of systematic studies on the parasitoids of aphids infesting alfalfa in Europe. Here we present an original and up-dated key for identification of both native and invasive Aphidiinae parasitoids attacking aphids that feed on alfalfa in Europe. In total, 20 species are keyed and illustrated with scanning electron micrographs and line drawings.

Re-visiting the Aphidius urticae s. str. group: re-description of Aphidius rubi Starý and A. silvaticus Starý (Hymenoptera: Braconidae: Aphidiinae).

Here we tested Aphidius urticae s. str. host-associated lineages from Microlophium carnosum (Buckton), Amphorophora rubi (Kaltenbach), Macrosiphum funestum (Macchiati) and Aulacorthum vaccinii Hille Ris Lambers with the barcoding region of the mitochondrial cytochrome oxidase subunit I gene used to analyse population differences and elucidate phylogenetic relationships between the separated taxa. This molecular marker has been shown to be the most informative molecular marker in resolving species complexes in aphidiine parasitoids. Analyses of the mitochondrial sequences revealed the existence of three clearly separated mitochondrial lineages of A. urticae s. str. group associated with: i) Macrosiphum funestum and Aulacorthum vaccinii aphid hosts, ii) Microlophium carnosum and iii) Amphorophora rubi. This corresponds to the initial descriptions of A. rubi, A. silvaticus and A. urticae and their aphid host associations prior to synonymization of A. rubi and A. silvaticus with A. urticae. On the other hand, significant evolutionary distances ranging from 2.3 to 9.2% between the three mitochondrial lineages were not accompanied by clear morphological differences. Therefore, re-descriptions of A. rubi and A. silvaticus are presented, together with their morphological differentiation in a key, as well as their phylogenetic relationships and genetical differentiation.

Genetic and morphological variation in sexual and asexual parasitoids of the genus Lysiphlebus - an apparent link between wing shape and reproductive mode.

BACKGROUND: Morphological divergence often increases with phylogenetic distance, thus making morphology taxonomically informative. However, transitions to asexual reproduction may complicate this relationship because asexual lineages capture and freeze parts of the phenotypic variation of the sexual populations from which they derive. Parasitoid wasps belonging to the genus Lysiphlebus Foerster (Hymenoptera: Braconidae: Aphidiinae) are composed of over 20 species that exploit over a hundred species of aphid hosts, including many important agricultural pests. Within Lysiphlebus, two genetically and morphologically well-defined species groups are recognised: the "fabarum" and the "testaceipes" groups. Yet within each group, sexual as well as asexual lineages occur, and in L. fabarum different morphs of unknown origin and status have been recognised. In this study, we selected a broad sample of specimens from the genus Lysiphlebus to explore the relationship between genetic divergence, reproductive mode and morphological variation in wing size and shape (quantified by geometric morphometrics). RESULTS: The analyses of mitochondrial and nuclear gene sequences revealed a clear separation between the "testaceipes" and "fabarum" groups of Lysiphlebus, as well as three well-defined phylogenetic lineages within the "fabarum" species group and two lineages within the "testaceipes" group. Divergence in wing shape was concordant with the deep split between the "testaceipes" and "fabarum" species groups, but within groups no clear association between genetic divergence and wing shape variation was observed. On the other hand, we found significant and consistent differences in the shape of the wing between sexual and asexual lineages, even when they were closely related. CONCLUSIONS: Mapping wing shape data onto an independently derived molecular phylogeny of Lysiphlebus revealed an association between genetic and morphological divergence only for the deepest phylogenetic split. In more recently diverged taxa, much of the variation in wing shape was explained by differences between sexual and asexual lineages, suggesting a mechanistic link between wing shape and reproductive mode in these parasitoid wasps.

Revision of the world Monoctonia Starý, parasitoids of gall aphids: taxonomy, distribution, host range, and phylogeny (Hymenoptera, Braconidae: Aphidiinae).

The present paper represents a contribution to the knowledge of the taxonomy of Monoctonia Starý aphid parasitoids obtained using the barcoding region of the mitochondrial COI gene. We discuss the phylogenetic position of the genus within the subtribe Monoctonina, redescribe known species, and describe Monoctonia japonica sp. n. from Japan in the association Pemphigus matsumurai Monzen/Populus maximowiczii. A key for species identification is provided. Also, we review and discuss the host records, origin, and geographical distribution of Monoctonia species. It is hypothesized that the genus Monoctonia evolved in Paleogene forests of the temperate (and subtropical) belt, most probably in the European part of the Mediterranean region, which is also the center of origin of their host plants. 

Interference of field evidence, morphology, and DNA analyses of three related Lysiphlebus aphid parasitoids (Hymenoptera: Braconidae: Aphidiinae).

This study provides evidence on integrating the morphological, field, and laboratory data, and application of the cytochrome oxidase subunit I (COI) barcoding gene to the three asexual or sexual Lysiphlebus spp., i.e., Lysiphlebus cardui (Marshall), Lysiphlebus confusus Tremblay and Eady and Lysiphlebus fabarum (Marshall) (Hymenoptera: Braconidae: Aphidiinae). New aphid- invasive plant association, Aphis fabae Scopoli (Hemipreta: Aphididae) on Impatiens glandulifera Royle, has been used in the same model area in the Czech Republic under the same sampling and rearing method for several consecutive years and throughout the season. For molecular identification of these three species, we used DNA sequences of the barcoding region of the mitochondrial COI gene. Although our results confirmed ecological and morphological differences among L. cardui, L. confusus, and L. fabarum, genetic analysis on the basis of COI mitochondrial barcoding gene does not support species status of the mentioned Lysiphlebus taxa. The level of morphological differentiation in these Lysiphlebus Förster species is in accordance with the usual species variability within subfamily Aphidiinae. However, it should be examined how appearance of asexual lineages affects the morphological or genetical variability.

Lysiphlebus orientalis (Hymenoptera, Braconidae), a new invasive aphid parasitoid in Europe - evidence from molecular markers.

We report the occurrence of Lysiphlebus orientalis in Serbia, an aphid parasitoid from the Far East that is new to Europe and has the potential to become invasive. Our finding based on morphological characters is confirmed by analyses of mitochondrial cytochrome oxidase subunit I sequences. An increase in number and an expansion of the host range were observed during field studies over the past two years, and it is determined that the current host range encompasses nine aphid hosts on 12 different host plants, forming 13 tri-trophic associations. A host range determined for European populations of L. orientalis appears wider compared with that in its Far Eastern native habitats where Aphis glycines Mats. is the sole known host. Moreover, it overlaps considerably with the host ranges of European parasitoids that play an important role in the natural control of pest aphids.