Mitogenomics of the Olive Seed Weevil, Anchonocranus oleae Marshall and Implications for Its Phylogenetic Position in Curculionidae.

Anchonocranus oleae Marshall (Coleoptera: Curculionidae) is a seed-feeding weevil native to southern Africa; its larvae are known to develop in the fruits of the African Wild Olive and, more rarely, cultivated olives. The species has been mainly found in the Western Cape province of South Africa, but it has remained in relative obscurity because it does not seem to represent a current threat to commercial olive production. As part of an ongoing effort to produce baseline genetic data for olive-associated entomofauna in South Africa, we generated reference DNA barcodes for A. oleae collected from wild and cultivated olives and sequenced its mitogenome for assessment of the phylogenetic position of the species in the family Curculionidae. The mitochondrial phylogeny estimate indicated that A. oleae shares a common ancestor with Elaidobius (tribe Derelomini), but a definite and close relationship to this tribe and the precise tribal placement of A. oleae in the subfamily Curculioninae could not be inferred due to the lack of representative mitogenomes of other relevant curculionine tribes and genera. This study will assist future work on the DNA-based species identification, genetic diversity, and phylogenetic position of the genus Anchonocranus and related taxa.

New Mitochondrial Gene Rearrangement in Psyttalia concolor, P. humilis and P. lounsburyi (Hymenoptera: Braconidae), Three Parasitoid Species of Economic Interest.

The family Braconidae consists mostly of specialized parasitoids, some of which hold potential in biocontrol of agricultural pests. Psyttalia concolor, Psyttalia humilis and Psyttalia lounsburyi are parasitoids associated with Bactrocera oleae, a major pest of cultivated olives. The native range of Psyttalia concolor is the Mediterranean, and P. humilis and P. lounsburyi are native to sub-Saharan Africa. This study reports the mitochondrial genomes of the three species, thus laying the foundation for mitogenomic analyses in the genus Psyttalia. Comparative mitogenomics within Braconidae showed a novel gene arrangement in Psyttalia in involving translocation and inversion of transfer RNA genes. The placement of Psyttalia in the subfamily Opiinae was well-supported, and the divergence between Psyttalia and its closest relative (Diachasmimorpha longicaudata) was at ~55 MYA [95% highest posterior density (HPD): 34-83 MYA]. Psyttalia lounsburyi occupied the most basal position among the three Psyttalia, having diverged from the other two species ~11 MYA (95% HPD: 6-17 MYA). Psyttalia concolor and P. humilis were recovered as sister species diverged at ~2 MYA (95% HPD: 1.1-3.6 MYA). This phylogeny combining new sequences and a set of 31 other cyclostomes and non-cyclostomes highlights the importance of a comprehensive taxonomic coverage of Braconidae mitogenomes to overcome the lack of robustness in the placement of several subfamilies.

Description of the sexual generation of Dryocosmus destefanii (Hymenoptera: Cynipidae: Cynipini) and disclosure of its life cycle.

The sexual generation of Dryocosmus destefanii Cerasa Melika, 2018 that emerges from galls on Q. suber L. in Italy is described for the first time, establishing its heterogonic life cycle. We provide observations on its distribution, illustration of adults and galls and information on its biology as supported by morphological and molecular data. An illustrated identification key to Western Palaearctic Dryocosmus species is also given.

Influence of Distance from the Host on Parasitisation by Psyttalia concolor (Szépligeti) (Hymenoptera: Braconidae).

The olive fruit fly, Bactrocera oleae, is considered the main olive pest worldwide, and has been the target of biological control programmes through the release of the braconid parasitoid Psyttalia concolor. Laboratory tests were performed to evaluate the influence of distance from the host on parasitisation, placing larvae of the substitute host Ceratitis capitata at seven distances (0, 0.5, 1, 1.5, 2, 2.5, 3 mm) and four different time periods (7, 15, 30, 60 min). Moreover, field collected olives of Ogliarola Barese cultivar infested by B. oleae were exposed to P. concolor females to confirm its ability to parasitise B. oleae in small olives. Psyttalia concolor oviposition was inhibited at 2.5 and 3 mm due to the ovipositor length of the parasitoid females (2.7 mm). Hosts were easily parasitised at distances between 0 and 1.5 mm. The thin fruit pulp (up to 3.5 mm) of field collected olives allowed the parasitisation to occur also in mature fruits. At the best combination distance/time (0 mm, 30 min), tests performed with different larvae/parasitoid female ratio showed an increasing emergence of P. concolor (from 20% to 57%) with larvae/parasitoid ratio increasing from 0.11 to 0.74. The results of the present study might optimise the mass rearing of P. concolor, through a proper setting of its parameters, such as the host/parasitoid ratio, exposure distances, and interaction time.

Barcoding of parasitoid wasps (Braconidae and Chalcidoidea) associated with wild and cultivated olives in the Western Cape of South Africa 1.

Wild and cultivated olives harbor and share a diversity of insects, some of which are considered agricultural pests, such as the olive fruit fly. The assemblage of olive-associated parasitoids and seed wasps is rich and specialized in sub-Saharan Africa, with native species possibly coevolving with their hosts. Although historical entomological surveys reported on the diversity of olive wasp species in the Western Cape Province of South Africa, no comprehensive study has been performed in the region in the molecular era. In this study, a dual approach combining morphological and DNA-based methods was used for the identification of adult specimens reared from olive fruits. Four species of Braconidae and six species of Chalcidoidea were identified, and DNA barcoding methodologies were used to investigate conspecificity among individuals, based on randomly selected representative specimens. Morphological identifications were congruent with DNA data, as NJ and ML trees correctly placed the sequences for each species either at the genus or species level, depending on the available taxa coverage, and genetic distances strongly supported conspecificity. No clear evidence of cryptic diversity was found. Overall seed infestation and parasitism rates were higher in wild olives compared to cultivated olives, and highest for Eupelmus spermophilus and Utetes africanus. These results can be used for early DNA-based detection of wasp larvae in olives and to further investigate the biology and ecology of these species.

The complete mitochondrial genome of Bactrocera biguttula (Bezzi) (Diptera: Tephritidae) and phylogenetic relationships with other Dacini.

Bactrocera biguttula is an African olive fruit fly that does not attack cultivated olives but rather develops in the fruits of wild species of Olea and Noronhia. The complete mitochondrial genome of an individual specimen was characterized in comparison to other Bactrocera. The phylogenetic relationships of B. biguttula with other Dacini were investigated, with special focus on B. oleae, an agricultural pest known to attack cultivated and wild olives. The sequence had a total length of 15,829 bp, and included the typical features of insect mitogenomes, similarly to the other Bactrocera analysed. Start codons included ATG, ATC, ATT, and TCG (in COI). The majority of stop codons (TAA) were fully encoded, whereas in some cases only TA or T were present. The complete sequence was biased towards A + T, with a positive AT-skew and a negative GC-skew. The predicted cloverleaf structure of tRNASer1 showed absence of the DHU arm, a common feature in insects and other Metazoans. Phylogenetic reconstruction showed that B. biguttula and B. oleae are sister species, having diverged from a common ancestor < 10 Myr ago. This result warrants future genomic comparisons between these two closely related species for investigating the specific adaptations to the different hosts.

Description of Dryocosmus destefanii new species (Hymenoptera: Cynipidae: Cynipini) from Quercus suber L. in Italy.

A new species, Dryocosmus destefanii Cerasa Melika n. sp. associated with a Cerris section oak, Quercus suber L., is described from Italy. Description, diagnosis, host associations and biology for the new species and an illustrated identification key to the Western Palaearctic Dryocosmus species are given. The description is supported by morphological and molecular data.

Revision of Afrotropical Dyscritobaeus Perkins, 1910 (Hymenoptera: Scelionidae).

A revision of Afrotropical species of Dyscritobaeus Perkins is presented with re-description of the four known species (D. bicolor O'Connor et Ashe, D. comitans Perkins, D. parvipennis (Dodd) and D. sulawensis Mineo, O'Connor et Ashe) and description of six new species (D. antananarivensis Tortorici et Caleca sp. nov., D. flavus Tortorici et Caleca sp. nov., D. kilimanjarensis Tortorici et Caleca sp. nov., D. madagascarensis Tortorici et Caleca sp. nov., D. ndokii Tortorici et Caleca sp. nov. and D. tanzaniensis Tortorici et Caleca sp. nov.). Dyscritobaeus cerosus is considered to be a junior synonym of D. comitans, D. hannibal is considered to be a junior synonym of D. sulawensis and D. maputanus is a junior synonym of D. parvipennis; so far these species are the only three Dyscritobaeus species that are widely distributed across four zoogeographical regions: Afrotropical, Australian, Oriental and Palearctic. Dyscritobaeus comitans and D. ndokii Tortorici et Caleca sp. nov. belong to the comitans-group, the other eight species belong to the orientalis-group. Dyscritobaeus antananarivensis Tortorici et Caleca sp. nov., D. madagascarensis Tortorici et Caleca sp. nov. and D. sulawensis are morphologically similar to D. orientalis bearing the specillum on T2; D. bicolor, D. flavus Tortorici et Caleca sp. nov., D. kilimanjarensis Tortorici et Caleca sp. nov., are morphologically similar to D. parvipennis; D. tanzaniensis Tortorici et Caleca sp. nov. shares the lack of a protruding metascutellum together with D. aspinosus Mineo, O'Connor et Ashe.Dyscritobaeus species are sexually dimorphic, particularly in the following features: the anterior and posterior fringes of the fore wing are longer in the female than in the male; the odontoid process, when present in females, is less developed or absent in the corresponding males; the specillum, when present in females, is less defined and smaller or absent in the corresponding males; the sculpture of the head is more evident in males than in females; and the first and second tergites are frequently lighter in males than in females.

Pathogenicity bioassays of isolates of Beauveria bassiana on Rhynchophorus ferrugineus.

BACKGROUND: The control of Rhynchophorus ferrugineus (Olivier), the main palm pest in the Mediterranean Basin, is problematic because of its biology and the current restrictions in many European countries on the use of chemical insecticides in urban areas. Entomopathogenic fungi have been studied as potential biological control agents, but information on their natural incidence is limited. Strains of Beauveria bassiana (Balsamo) Vuillemin were isolated from symptomatic insects collected on dead palms, and their pathogenicity against different instars of R. ferrugineus was evaluated in the laboratory. RESULTS: The overall percentage of infected insects found in Canary palms was 7%. In laboratory bioassays, hatching of eggs treated with three different isolates of B. bassiana was 41.2, 26.8 and 29.9%, significantly lower than the control (62.4%). Larvae and adults were treated with a single isolate in two ways: spraying each insect with a conidial suspension or feeding them with fruit portions previously immersed in the same conidial suspension. At the end of the two trials, the mortality of treated larvae was 88 and 92%, and the mean survival time was 10.4 and 11.8 days, significantly different from the control, where no insect died during the trials. Mortality and survival time recorded in either trial on adults did not significantly differ between treatment and control. CONCLUSION: This study shows that the pathogenicity of wild isolates of B. bassiana differs among the tested R. ferrugineus instars. The low mortality of treated adults supports their use as vectors of B. bassiana as a potential tool for reducing R. ferrugineus populations.