Morphological phylogeny and revision of Sycophila and Ficomila (Hymenoptera: Chalcidoidea, Eurytomidae) associated with Afrotropical fig trees (Moraceae, Ficus).

Eurytomidae (Chalcidoidea) species associated with fig trees (Ficus) are still poorly documented. A phylogenetic analysis of 63 morphological characters was conducted to revise Afrotropical species of Sycophila Walker and Ficomila Bouek associated with fig trees. Based on our results, which also included Palaearctic species of Sycophila, three subgenera of Sycophila are proposed: Sycophila s. str., Tineomyza Rondani stat. rev. and a new subgenus, Ficorytoma subgen. n. Lotfalizadeh & Rasplus. Ficomila is maintained as a valid genus. Thirty-six species are also newly described by Lotfalizadeh & Rasplus, namely Ficomila artocarpoides sp. n., F. carolae sp. n., F. bouceki sp. n., F. comptoni sp. n., F. flava sp. n., F. gabonensis sp. n., F. gibba sp. n., F. guinensis sp. n., F. sinai sp. n., F. tanzanica sp. n., F. vannoorti sp. n., Sycophila (Tineomyza) beninensis sp. n., S. (T.) busseicola sp. n., S. (T.) glumosae sp. n., S. (T.) luteacola sp. n., S. (T.) maldesi sp. n., S. (T.) minuta sp. n., S. (T.) platygastra sp. n., S. (T.) risbeci sp. n., S. (T.) wiebesi sp. n., S. (T.) zebrogastra sp. n., S. (Ficorytoma) delvarei sp. n., S. (F.) persicae sp. n., S. (Sycophila) annae sp. n., S. (S.) bidentata sp. n., S. (S.) longiflagellata sp. n., S. (S.) chirindensis sp. n., S. (S.) ficophila sp. n., S. (S.) fusca sp. n., S. (S.) lasallei sp. n., S. (S.) macrospermae sp. n., S. (S.) meridionalis sp. n., S. (S.) nigra sp. n., S. (S.) nigriterga sp. n., S. (S.) suricola sp. n., S. (S.) zebra sp. n.. Seven described species of Sycophila are reclassified in the subgenus Tineomyza: Sycophila (Tineomyza) flaviclava Bouek, 1981; S. (T.) modesta Bouek, 1981; S. (T.) naso Bouek, 1981; S. (T.) punctum Bouek, 1981; S. (T.) ruandensis Risbec, 1957; S. (T.) sessilis Bouek, 1981 and S. (T.) setulosa Zerova, 2009. After this revision, Ficomila and Sycophila include 46 species associated with Afrotropical Ficus. Detailed morphological descriptions and illustrations as well as identification keys are provided for the subgenera, species-groups and species of Ficomila and Sycophila associated with Afrotropical fig trees; their host fig association and relative lack of host-specificity are also discussed.

New genera of Afrotropical Chalcidoidea (Hymenoptera: Cerocephalidae, Epichrysomallidae, Pirenidae and Pteromalidae).

As a preliminary step towards the development of a key to genera of several families of Afrotropical Chalcidoidea, seven new genera in four families are described: Cerocephalidae-Milokoa Mitroiu, gen. nov. (type species: Milokoa villemantae Mitroiu, sp. nov.); Epichrysomallidae-Delvareus Rasplus, Mitroiu & van Noort, gen. nov. (type species: Delvareus dicranostylae Rasplus, Mitroiu & van Noort, sp. nov.); Pirenidae-Afrothopus Mitroiu, gen. nov. (type species: Afrothopus georgei Mitroiu, sp. nov.); Pteromalidae-Kerangania Mitroiu, gen. nov. (type species: Kerangania nuda Mitroiu, sp. nov.), Pilosalis Mitroiu, Rasplus & van Noort, gen. nov. (type species: Pilosalis barbatulus Mitroiu, sp. nov.), Scrobesia Mitroiu & Rasplus, gen. nov. (type species: Scrobesia acutigaster Mitroiu & Rasplus, sp. nov.), and Spiniclava Mitroiu & Rasplus, gen. nov. (type species: Spiniclava baaiensis Mitroiu & Rasplus, sp. nov.). Additionally, the following new species are described: Pilosalis bouceki Mitroiu & Rasplus, sp. nov., Pilosalis eurys Mitroiu & van Noort, sp. nov., Pilosalis minutus Mitroiu, sp. nov., Pilosalis platyscapus Mitroiu, Rasplus & van Noort, sp. nov., Scrobesia pondo Mitroiu, sp. nov., and Spiniclava setosa Mitroiu, sp. nov. All taxa are illustrated and the relationships with similar taxa are discussed. For each non-monotypic genus a key to species is provided.

The Chalcidoidea bush of life: evolutionary history of a massive radiation of minute wasps.

Chalcidoidea are mostly parasitoid wasps that include as many as 500 000 estimated species. Capturing phylogenetic signal from such a massive radiation can be daunting. Chalcidoidea is an excellent example of a hyperdiverse group that has remained recalcitrant to phylogenetic resolution. We combined 1007 exons obtained with Anchored Hybrid Enrichment with 1048 ultra-conserved elements (UCEs) for 433 taxa including all extant families, >95% of all subfamilies, and 356 genera chosen to represent the vast diversity of the superfamily. Going back and forth between the molecular results and our collective knowledge of morphology and biology, we detected bias in the analyses that was driven by the saturation of nucleotide data. Our final results are based on a concatenated analysis of the least saturated exons and UCE datasets (2054 loci, 284 106 sites). Our analyses support an expected sister relationship with Mymarommatoidea. Seven previously recognized families were not monophyletic, so support for a new classification is discussed. Natural history in some cases would appear to be more informative than morphology, as illustrated by the elucidation of a clade of plant gall associates and a clade of taxa with planidial first-instar larvae. The phylogeny suggests a transition from smaller soft-bodied wasps to larger and more heavily sclerotized wasps, with egg parasitism as potentially ancestral for the entire superfamily. Deep divergences in Chalcidoidea coincide with an increase in insect families in the fossil record, and an early shift to phytophagy corresponds with the beginning of the "Angiosperm Terrestrial Revolution". Our dating analyses suggest a middle Jurassic origin of 174 Ma (167.3-180.5 Ma) and a crown age of 162.2 Ma (153.9-169.8 Ma) for Chalcidoidea. During the Cretaceous, Chalcidoidea may have undergone a rapid radiation in southern Gondwana with subsequent dispersals to the Northern Hemisphere. This scenario is discussed with regard to knowledge about the host taxa of chalcid wasps, their fossil record and Earth's palaeogeographic history.

Echoes of ancient introgression punctuate stable genomic lineages in the evolution of figs.

Studies investigating the evolution of flowering plants have long focused on isolating mechanisms such as pollinator specificity. Some recent studies have proposed a role for introgressive hybridization between species, recognizing that isolating processes such as pollinator specialization may not be complete barriers to hybridization. Occasional hybridization may therefore lead to distinct yet reproductively connected lineages. We investigate the balance between introgression and reproductive isolation in a diverse clade using a densely sampled phylogenomic study of fig trees (Ficus, Moraceae). Codiversification with specialized pollinating wasps (Agaonidae) is recognized as a major engine of fig diversity, leading to about 850 species. Nevertheless, some studies have focused on the importance of hybridization in Ficus, highlighting the consequences of pollinator sharing. Here, we employ dense taxon sampling (520 species) throughout Moraceae and 1,751 loci to investigate phylogenetic relationships and the prevalence of introgression among species throughout the history of Ficus. We present a well-resolved phylogenomic backbone for Ficus, providing a solid foundation for an updated classification. Our results paint a picture of phylogenetically stable evolution within lineages punctuated by occasional local introgression events likely mediated by local pollinator sharing, illustrated by clear cases of cytoplasmic introgression that have been nearly drowned out of the nuclear genome through subsequent lineage fidelity. The phylogenetic history of figs thus highlights that while hybridization is an important process in plant evolution, the mere ability of species to hybridize locally does not necessarily translate into ongoing introgression between distant lineages, particularly in the presence of obligate plant-pollinator relationships.

Endoparasitoid lifestyle promotes endogenization and domestication of dsDNA viruses.

The accidental endogenization of viral elements within eukaryotic genomes can occasionally provide significant evolutionary benefits, giving rise to their long-term retention, that is, to viral domestication. For instance, in some endoparasitoid wasps (whose immature stages develop inside their hosts), the membrane-fusion property of double-stranded DNA viruses have been repeatedly domesticated following ancestral endogenizations. The endogenized genes provide female wasps with a delivery tool to inject virulence factors that are essential to the developmental success of their offspring. Because all known cases of viral domestication involve endoparasitic wasps, we hypothesized that this lifestyle, relying on a close interaction between individuals, may have promoted the endogenization and domestication of viruses. By analyzing the composition of 124 Hymenoptera genomes, spread over the diversity of this clade and including free-living, ecto, and endoparasitoid species, we tested this hypothesis. Our analysis first revealed that double-stranded DNA viruses, in comparison with other viral genomic structures (ssDNA, dsRNA, ssRNA), are more often endogenized and domesticated (that is, retained by selection) than expected from their estimated abundance in insect viral communities. Second, our analysis indicates that the rate at which dsDNA viruses are endogenized is higher in endoparasitoids than in ectoparasitoids or free-living hymenopterans, which also translates into more frequent events of domestication. Hence, these results are consistent with the hypothesis that the endoparasitoid lifestyle has facilitated the endogenization of dsDNA viruses, in turn, increasing the opportunities of domestications that now play a central role in the biology of many endoparasitoid lineages.

Novel Gene Rearrangement Pattern in Pachycrepoideus vindemmiae Mitochondrial Genome: New Gene Order in Pteromalidae (Hymenoptera: Chalcidoidea).

The mitochondrial genomes of Muscidifurax similadanacus, M. sinesensilla, Nasonia vitripennis, and Pachycrepoideus vindemmiae were sequenced to better understand the structural evolution of Pteromalidae mitogenomes. These newly sequenced mitogenomes all contained 37 genes. Nucleotide composition was AT-biased and the majority of the protein-coding genes exhibited a negative AT skew. All 13 protein-coding genes (PCGs) initiated with the standard start codon of ATN, excepted for nad1 of N. vitripennis, which started with TTG, and terminated with a typical stop codon TAA/TAG or an incomplete stop codon T. All transfer RNA (tRNA) genes were predicted to fold into the typical clover-leaf secondary structures, except for trnS1, which lacks the DHU arm in all species. In P. vindemmiae, trnR and trnQ lack the DHU arm and TΨC arm, respectively. Although most genes evolved under a strong purifying selection, the Ka/Ks value of the atp8 gene of P. vindemmiae was greater than 1, indicating putative positive selection. A novel transposition of trnR in P. vindemmiae was revealed, which was the first of this kind to be reported in Pteromalidae. Two kinds of datasets (PCG12 and AA) and two inference methods (maximum likelihood and Bayesian inference) were used to reconstruct a phylogenetic hypothesis for the newly sequenced mitogenomes of Pteromalidae and those deposited in GenBank. The topologies obtained recovered the monophyly of the three subfamilies included. Pachyneurinae and Pteromalinae were recovered as sister families, and both appeared sister to Sycophaginae. The pairwise breakpoint distances of mitogenome rearrangements were estimated to infer phylogeny among pteromalid species. The topology obtained was not totally congruent with those reconstructed using the ML and BI methods.

Key innovations and the diversification of Hymenoptera.

The order Hymenoptera (wasps, ants, sawflies, and bees) represents one of the most diverse animal lineages, but whether specific key innovations have contributed to its diversification is still unknown. We assembled the largest time-calibrated phylogeny of Hymenoptera to date and investigated the origin and possible correlation of particular morphological and behavioral innovations with diversification in the order: the wasp waist of Apocrita; the stinger of Aculeata; parasitoidism, a specialized form of carnivory; and secondary phytophagy, a reversal to plant-feeding. Here, we show that parasitoidism has been the dominant strategy since the Late Triassic in Hymenoptera, but was not an immediate driver of diversification. Instead, transitions to secondary phytophagy (from parasitoidism) had a major influence on diversification rate in Hymenoptera. Support for the stinger and the wasp waist as key innovations remains equivocal, but these traits may have laid the anatomical and behavioral foundations for adaptations more directly associated with diversification.

Environmental factors driving the abundance of Philaenus spumarius in mesomediterranean habitats of Corsica (France).

Philaenus spumarius (Ps) is considered the main insect vector of the bacterium Xylella fastidiosa (Xf) in Europe. As such, it is a key actor of the Xf pathosystem on which surveillance and management strategies could be implemented. Although research effort has increased in the past years, the ecological factors shaping Ps abundance and distribution across landscapes are still poorly known in most regions of Europe. We selected 64 plots of 500m2 in Corsican semi-natural habitats in which we sampled nymphs and adults of Ps during three years. While local or surrounding vegetation structure (low or high scrubland) had little effect on Ps abundance, we highlighted a positive relationship between Ps abundance and the density of Cistus monspeliensis in the plots. We also found larger populations of Ps in cooler and moister plots. The pattern of host association highlighted here is unique, which calls for more studies on the ecology of Ps in Europe, to help designing surveillance and management strategy for Xf.

Climate change and the potential distribution of the glassy-winged sharpshooter (Homalodisca vitripennis), an insect vector of Xylella fastidiosa.

Biological invasions represent a major threat for biodiversity and agriculture. Despite efforts to restrict the spread of alien species, preventing their introduction remains the best strategy for an efficient control. In that context preparedness of phytosanitary authorities is very important and estimating the geographical range of alien species becomes a key information. The present study investigates the potential geographical range of the glassy-winged sharpshooter (Homalodisca vitripennis), a very efficient insect vector of Xylella fastidiosa, one of the most dangerous plant-pathogenic bacteria worldwide. We use species distribution modeling (SDM) to analyse the climate factors driving the insect distribution and we evaluate its potential distribution in its native range (USA) and in Europe according to current climate and different scenarios of climate change: 6 General Circulation Models (GCM), 4 shared socioeconomic pathways of gas emission and 4 time periods (2030, 2050, 2070, 2090). The first result is that the climate conditions of the European continent are suitable to the glassy-winged sharpshooter, in particular around the Mediterranean basin where X. fastidiosa is present. Projections according to future climate conditions indicate displacement of climatically suitable areas towards the north in both North America and Europe. Globally, suitable areas will decrease in North America and increase in Europe in the coming decades. SDM outputs vary according to the GCM considered and this variability indicated areas of uncertainty in the species potential range. Both potential distribution and its uncertainty associated to future climate projections are important information for improved preparedness of phytosanitary authorities.

Potential Parasitoids for Biocontrol of the Ber Fruit Fly, Carpomya vesuviana Costa (Diptera: Tephritidae).

The ber fruit fly (BFF), Carpomya vesuviana Costa, 1854 (Diptera: Tephritidae), is an important key pest of the jujube, Ziziphus jujuba Miller. The main control measures against this pest are to use chemical control, but the first survey for its natural enemies was performed in Iran. Here, we report eight species of parasitic wasps of the BFF from five different families. The family Eurytomidae with three species, the families Pteromalidae and Mutillidae with two species each, and the families Braconidae and Diapriidae with one species each are associated with different immature stages of the BFF, of which Eurytoma pineticola Zerova (Eurytomidae) and Cyrtoptyx lichtensteini (Masi) (Pteromalidae) were the most abundant parasitoid species. Fopius carpomyiae (Silvestri,) was not reared on BFF on the jujube during this survey, but it was reported on Ziziphus spina-christi (L.) with a high parasitism rate. Therefore, it seems to be the most important parasitoid of BFF in Iran. The parasitoid community of BFF in Khorasan, Eastern Iran, is reviewed, and an identification key to these species is proposed.

Vectors as Sentinels: Rising Temperatures Increase the Risk of Xylella fastidiosa Outbreaks.

Global change is expected to modify the threat posed by pathogens to plants. However, little is known regarding how a changing climate will influence the epidemiology of generalist vector-borne diseases. We developed a high-throughput screening method to test for the presence of a deadly plant pathogen, Xylella fastidiosa, in its insect vectors. Then, using data from a four-year survey in climatically distinct areas of Corsica (France), we demonstrated a positive correlation between the proportion of vectors positive to X. fastidiosa and temperature. Notably, a higher prevalence corresponded with milder winters. Our projections up to 2100 indicate an increased risk of outbreaks. While the proportion of vectors that carry the pathogen should increase, the climate conditions will remain suitable for the bacterium and its main vector, with possible range shifts towards a higher elevation. Besides calling for research efforts to limit the incidence of plant diseases in the temperate zone, this work reveals that recent molecular technologies could and should be used for massive screening of pathogens in vectors to scale-up surveillance and management efforts.

Ultra-Conserved Elements and morphology reciprocally illuminate conflicting phylogenetic hypotheses in Chalcididae (Hymenoptera, Chalcidoidea).

Recent technical advances combined with novel computational approaches have promised the acceleration of our understanding of the tree of life. However, when it comes to hyperdiverse and poorly known groups of invertebrates, studies are still scarce. As published phylogenies will be rarely challenged by future taxonomists, careful attention must be paid to potential analytical bias. We present the first molecular phylogenetic hypothesis for the family Chalcididae, a group of parasitoid wasps, with a representative sampling (144 ingroups and seven outgroups) that covers all described subfamilies and tribes, and 82% of the known genera. Analyses of 538 Ultra-Conserved Elements (UCEs) with supermatrix (RAxML and IQTREE) and gene tree reconciliation approaches (ASTRAL, ASTRID) resulted in highly supported topologies in overall agreement with morphology but reveal conflicting topologies for some of the deepest nodes. To resolve these conflicts, we explored the phylogenetic tree space with clustering and gene genealogy interrogation methods, analyzed marker and taxon properties that could bias inferences and performed a thorough morphological analysis (130 characters encoded for 40 taxa representative of the diversity). This joint analysis reveals that UCEs enable attainment of resolution between ancestry and convergent/divergent evolution when morphology is not informative enough, but also shows that a systematic exploration of bias with different analytical methods and a careful analysis of morphological features is required to prevent publication of artifactual results. We highlight a GC content bias for maximum-likelihood approaches, an artifactual mid-point rooting of the ASTRAL tree and a deleterious effect of high percentage of missing data (>85% missing UCEs) on gene tree reconciliation methods. Based on the results we propose a new classification of the family into eight subfamilies and ten tribes that lay the foundation for future studies on the evolutionary history of Chalcididae.

Exploring systematic biases, rooting methods and morphological evidence to unravel the evolutionary history of the genus Ficus (Moraceae).

Despite many attempts in the Sanger sequencing era, the phylogeny of fig trees remains unresolved, which limits our ability to analyze the evolution of key traits that may have contributed to their evolutionary and ecological success. We used restriction-site-associated DNA sequencing (c. 420 kb) and 102 morphological characters to elucidate the relationships between 70 species of Ficus. To increase phylogenetic information for higher-level relationships, we targeted conserved regions and assembled paired reads into long loci to enable the retrieval of homologous loci in outgroup genomes. We compared morphological and molecular results to highlight discrepancies and reveal possible inference bias. For the first time, we recovered a monophyletic subgenus Urostigma (stranglers) and a clade with all gynodioecious Ficus. However, we show, with a new approach based on iterative principal component analysis, that it is not (and will probably never be) possible to homogenize evolutionary rates and GC content for all taxa before phylogenetic inference. Four competing positions for the root of the molecular tree are possible. The placement of section Pharmacosycea as sister to other fig trees is not supported by morphological data and considered a result of a long-branch attraction artefact to the outgroups. Regarding morphological features and indirect evidence from the pollinator tree of life, the topology that divides Ficus into monoecious versus gynodioecious species appears most plausible. It seems most likely that the ancestor of fig trees was a freestanding tree and active pollination is inferred as the ancestral state, contrary to previous hypotheses. However, ambiguity remains on the ancestral breeding system. Despite morphological plasticity, we advocate restoring a central role to morphology in our understanding of the evolution of Ficus, as it can help detect systematic errors that appear more pronounced with larger molecular datasets.

Evolution, systematics and historical biogeography of sand flies of the subgenus Paraphlebotomus (Diptera, Psychodidae, Phlebotomus) inferred using restriction-site associated DNA markers.

Phlebotomine sand flies are the main natural vectors of Leishmania, which cause visceral and tegumentary tropical diseases worldwide. However, their taxonomy and evolutionary history remain poorly studied. Indeed, as for many human disease vectors, their small size is a challenge for morphological and molecular works. Here, we successfully amplified unbiased copies of whole genome to sequence thousands of restriction-site associated DNA (RAD) markers from single specimens of phlebotomines. RAD markers were used to infer a fully resolved phylogeny of the subgenus Paraphlebotomus (11 species + 5 outgroups, 32 specimens). The subgenus was not recovered as monophyletic and we describe a new subgenus Artemievus subg. nov. Depaquit for Phlebotomus alexandri. We also confirm the validity of Ph. riouxi which is reinstated as valid species. Our analyses suggest that Paraphlebotomus sensu nov. originated ca 12.9-8.5 Ma and was possibly largely distributed from peri-Mediterranean to Irano-Turanian regions. Its biogeographical history can be summarized into three phases: i) a first split between Ph. riouxi + Ph. chabaudi and other species that may have resulted from the rise of the Saharan belt ca 8.5 Ma; ii) a Messinian vicariant event (7.3-5.3 Ma) during which the prolonged drought could have resulted in the divergence of main lineages; iii) a recent radiation event (3-2 Ma) that correspond to cycles of wet and dry periods in the Middle East and the East African subregions during the Pleistocene. Interestingly these cycles are also hypothetical drivers of the diversification of rodents, in the burrows of which Paraphlebotomus larvae develop. By meeting the challenge of sequencing pangenomics markers from single, minute phlebotomines, this work opens new avenues for improving our understanding of the epidemiology of leishmaniases and possibly other human diseases transmitted by arthropod vectors.

Molecular mechanisms of mutualistic and antagonistic interactions in a plant-pollinator association.

Many insects metamorphose from antagonistic larvae into mutualistic adult pollinators, with reciprocal adaptation leading to specialized insect-plant associations. It remains unknown how such interactions are established at molecular level. Here we assemble high-quality genomes of a fig species, Ficus pumila var. pumila, and its specific pollinating wasp, Wiebesia pumilae. We combine multi-omics with validation experiments to reveal molecular mechanisms underlying this specialized interaction. In the plant, we identify the specific compound attracting pollinators and validate the function of several key genes regulating its biosynthesis. In the pollinator, we find a highly reduced number of odorant-binding protein genes and an odorant-binding protein mainly binding the attractant. During antagonistic interaction, we find similar chemical profiles and turnovers throughout the development of galled ovules and seeds, and a significant contraction of detoxification-related gene families in the pollinator. Our study identifies some key genes bridging coevolved mutualists, establishing expectations for more diffuse insect-pollinator systems.

Ooctonus vulgatus (Hymenoptera, Mymaridae), a potential biocontrol agent to reduce populations of Philaenus spumarius (Hemiptera, Aphrophoridae) the main vector of Xylella fastidiosa in Europe.

As a vector of Xylella fastidiosa (Wells, 1987) in Europe, the meadow spittlebug Philaenus spumarius (Linnaeus, 1758) (Hemiptera, Aphrophoridae) is a species of major concern. Therefore, tools and agents to control this ubiquitous insect that develops and feeds on hundreds of plant species are wanted. We conducted a field survey of P. spumarius eggs in Corsica and provide a first report of Ooctonus vulgatus Haliday, 1833 (Hymenoptera, Mymaridae) as a potential biocontrol agent of P. spumarius in Europe. To allow species identification, we summarized the main characters distinguishing O. vulgatus from other European species of Ooctonus and generated COI DNA barcodes. Parasitism rates were variable in the four localities included in the survey but could reach 69% (for an average number of eggs that hatched per locality of 109). Based on the geographic occurrences of O. vulgatus obtained from the literature, we calibrated an ecological niche model to assess its potential distribution in the Holarctic. Obviously, several questions need to be addressed to determine whether O. vulgatus could become an effective biocontrol agent of P. spumarius in Europe. So far, O. vulgatus has been reared only from P. spumarius eggs, but its exact host-range should be evaluated to ensure efficiency and avoid non-target effect. The top-down impact of the parasitoid on vector populations should also be assessed on large data sets. Finally, the feasibility of mass rearing should be tested. We hope this report serves as a starting point to initiate research on this parasitoid wasp to assess whether it could contribute to reduce the spread and impact of X. fastidiosa in Europe.

Tetramesa amica and its parasitoid Eurytoma amicophaga (Hymenoptera, Eurytomidae): two new species associated with medusahead, Taeniatherum caput-medusae (Poaceae).

Medusahead, Taeniatherum caput-medusae (Poales: Poaceae), is an annual grass native to central Asia and the Mediterranean region. It is a noxious, invasive weed in much of western North America. During field explorations carried out in Greece in 2017, the new phytophagous eurytomid Tetramesa amica Lotfalizadeh, sp. nov. and its parasitoid Eurytoma amicophaga Lotfalizadeh, sp. nov., also new to science, were recorded for the first time on medusahead. These new species are described and characters that enable to recognize them from their closest relatives are summarized. Tetramesa species are generally species-specific gall-inducers. They induce damages that may have a significant impact on the physiology of infested plants by reducing the productivity of flowering heads and seed weight. Based on these data, T. amica Lotfalizadeh, sp. nov. is currently being investigated as a candidate biological control agent of medusahead.

Xylella fastidiosa: climate suitability of European continent.

The bacterium Xylella fastidiosa (Xf) is a plant endophyte native to the Americas that causes diseases in many crops of economic importance (grapevine, Citrus, Olive trees etc). Xf has been recently detected in several regions outside of its native range including Europe where little is known about its potential geographical expansion. We collected data documenting the native and invaded ranges of the Xf subspecies fastidiosa, pauca and multiplex and fitted bioclimatic species distribution models (SDMs) to assess the potential climate suitability of European continent for those pathogens. According to model predictions, the currently reported distribution of Xf in Europe is small compared to the large extent of climatically suitable areas. The regions at high risk encompass the Mediterranean coastal areas of Spain, Greece, Italy and France, the Atlantic coastal areas of France, Portugal and Spain as well as the southwestern regions of Spain and lowlands in southern Italy. The extent of predicted climatically suitable conditions for the different subspecies are contrasted. The subspecies multiplex, and to a certain extent the subspecies fastidiosa, represent a threat to most of Europe while the climatically suitable areas for the subspecies pauca are mostly limited to the Mediterranean basin. These results provide crucial information for the design of a spatially informed European-scale integrated management strategy, including early detection surveys in plants and insect vectors and quarantine measures.

Restriction-site associated DNA markers provide new insights into the evolutionary history of the bark beetle genus Dendroctonus.

The bark beetle genus Dendroctonus contains some of the most economically important pests of conifers worldwide. Despite many attempts, there is no agreement today on the phylogenetic relationships within the genus, which limits our understanding of its evolutionary history. Here, using restriction-site associated DNA (RAD) markers from 70 specimens representing 17 species (85% of the known diversity) we inferred the phylogeny of the genus, its time of origin and biogeographic history, as well as the evolution of key ecological traits (host plants, larval behavior and adults' attack strategies). For all combinations of tested parameters (from 6444 to 23,570 RAD tags analyzed), the same, fully resolved topology was inferred. Our analyses suggest that the most recent common ancestor (mrca) of all extant Dendroctonus species was widely distributed across eastern Palearctic and western Nearctic during the early Miocene, from where species dispersed to other Holarctic regions. A first main inter-continental vicariance event occurred during early Miocene isolating the ancestors of D. armandi in the Palearctic, which was followed by the radiation of the main Dendroctonus lineages in North America. During the Late Miocene, the ancestor of the 'rufipennis' species group colonized north-east Palearctic regions from western North America, which was followed by a second main inter-continental vicariance event isolating Pleistocene populations in Asia (D. micans) and western North America (D. murrayanae and D. punctatus). The present study supports previous hypotheses explaining intercontinental range disjunctions across the Northern Hemisphere by the fragmentation of a continuous distribution due to climatic cooling, host range fragmentation and geological changes during the late Cenozoic. The reconstruction of ancestral ecological traits indicates that the mrca bored individual galleries and mass attacked the boles of pines. The gregarious feeding behavior of the larvae as well as the individual attack of the base of trees have apparently independently evolved twice in North America (in the 'rufipennis' and the 'valens' species groups), which suggests a higher adaptive potential than previously thought and may be of interest for plant protection and biodiversity conservation in a rapidly changing world.

Optimized DNA extraction and library preparation for minute arthropods: Application to target enrichment in chalcid wasps used for biocontrol.

Target enrichment is increasingly used for genotyping of plant and animal species or to better understand the evolutionary history of important lineages through the inference of statistically robust phylogenies. Limitations to routine target enrichment are both the complexity of current protocols and low input DNA quantity. Thus, working with tiny organisms such as microarthropods can be challenging. Here, we propose easy to set up optimizations for DNA extraction and library preparation prior to target enrichment. Prepared libraries were used to capture 1,432 ultraconserved elements (UCEs) from microhymenoptera (Chalcidoidea), which are among the tiniest insects on Earth and the most commercialized worldwide for biological control purposes. Results show no correlation between input DNA quantities (1.8-250 ng, 0.4 ng with an extra whole genome amplification step) and the number of sequenced UCEs on an Illumina MiSeq. Phylogenetic inferences highlight the potential of UCEs to solve relationships within the families of chalcid wasps, which has not been achieved so far. The protocol (library preparation + target enrichment) allows processing 96 specimens in five working days, by a single person, without requiring the use of expensive robotic molecular biology platforms, which could help to generalize the use of target enrichment for minute specimens.