Maríndia Deprá

The first records of the invasive pest Drosophila suzukii in the South American continent

This paper presents the first report of the occurrence of Drosophila suzukii (Diptera: Drosophilidae) in South America. Several samples were collected during various seasons in 2012 and 2013, and a significant number of individuals were obtained and identified. Based on the data recorded during each sampling campaign, including the locality, temperature, time of year, and number of individuals collected, several conclusions were drawn about the development of this invasion in southern Brazil.

Deciphering the routes of invasion of Drosophila suzukii by means of ABC random forest

Abstract Deciphering invasion routes from molecular data is crucial to understanding biological invasions, including identifying bottlenecks in population size and admixture among distinct populations. Here, we unravel the invasion routes of the invasive pest Drosophila suzukii using a multi-locus microsatellite dataset (25 loci on 23 worldwide sampling locations). To do this, we use approximate Bayesian computation (ABC), which has improved the reconstruction of invasion routes, but can be computationally expensive. We use our study to illustrate the use of a new, more efficient, ABC method, ABC random forest (ABC-RF) and compare it to a standard ABC method (ABC-LDA). We find that Japan emerges as the most probable source of the earliest recorded invasion into Hawaii. Southeast China and Hawaii together are the most probable sources of populations in western North America, which then in turn served as sources for those in eastern North America. European populations are genetically more homogeneous than North American populations, and their most probable source is northeast China, with evidence of limited gene flow from the eastern US as well. All introduced populations passed through bottlenecks, and analyses reveal five distinct admixture events. These findings can inform hypotheses concerning how this species evolved between different and independent source and invasive populations. Methodological comparisons indicate that ABC-RF and ABC-LDA show concordant results if ABC-LDA is based on a large number of simulated datasets but that ABC-RF out-performs ABC-LDA when using a comparable and more manageable number of simulated datasets, especially when analyzing complex introduction scenarios.

Mar, a MITE family of hAT transposons in Drosophila.

BackgroundMiniature inverted-repeat transposable elements (MITEs) are short, nonautonomous DNA elements flanked by subterminal or terminal inverted repeats (TIRs) with no coding capacity. MITEs were originally recognized as important components of plant genomes, where they can attain extremely high copy numbers, and are also found in several animal genomes, including mosquitoes, fish and humans. So far, few MITEs have been described in Drosophila.ResultsHerein we describe the distribution and evolution of Mar, a MITE family of hAT transposons, in Drosophilidae species. In silico searches and PCR screening showed that Mar distribution is restricted to the willistoni subgroup of the Drosophila species, and a phylogenetic analysis of Mar indicates that this element may have originated prior to the diversification of these species. Most of the Mar copies in D. willistoni present conserved target site duplications and TIRs, indicating recent mobilization of these sequences. We also identified relic copies of potentially full-length Mar transposon in D. tropicalis and D. willistoni. The phylogenetic relationship among transposases from the putative full-length Mar and other hAT superfamily elements revealed that Mar is placed into the recently determined Buster group of hAT transposons.ConclusionOn the basis of the obtained data, we can suggest that the origin of these Mar MITEs occurred before the subgroup willistoni speciation, which started about 5.7 Mya. The Mar relic transposase existence indicates that these MITEs originated by internal deletions and suggests that the full-length transposon was recently functional in D. willistoni, promoting Mar MITEs mobilization.

hosimary: a new hAT transposon group involved in horizontal transfer.

A PCR screening approach was used to search for sequences homologous to a previously described hAT transposon found in Drosophilasimulans and Drosophila sechellia, named here as hosimary. In this study, 52 Drosophilidae species were analyzed and these sequences seem to be restricted to some species of the melanogaster group and Zaprionus indianus. These species present variable number of copies and most of those appear to be putatively encoding. The high hosimary sequences similarity among different species and the patchy distribution presented by this transposon strongly support the hypothesis that hosimary was horizontally transferred between the melanogaster group species and Z. indianus.

Reevaluating the infection status by the Wolbachia endosymbiont in Drosophila Neotropical species from the willistoni subgroup

Infections by the endosymbiotic bacterium Wolbachia developed a rapid global expansion within Old World Drosophila species, ultimately infecting also Neotropical species. In this sense, screenings are necessary to characterize new variants of Wolbachia or new hosts, and also in order to map the dynamics of already known infections. In this paper, we performed a double screening approach that combined Dot-blot and PCR techniques in order to reevaluate the infection status by Wolbachia in species from the willistoni subgroup of Drosophila. Genomic DNA from isofemale lines descendent from females collected in the Amazonian Rainforest (n=91) were submitted to Dot-blot, and were positive for Wolbachia, producing a gradient of hybridization signals, suggesting different infection levels, which was further confirmed through quantitative PCR. Samples with a strong signal in the Dot-blot easily amplified in the wsp-PCR, unlike most of the samples with a medium to weak signal. It was possible to molecularly characterize three Drosophila equinoxialis isofemale lines that were found to be infected in a low density by a wMel-like Wolbachia strain, which was also verified in a laboratory line of Drosophila paulistorum Amazonian. We also found Drosophila tropicalis to be infected with the wAu strain and a Drosophila paulistorum Andean-Brazilian semispecies laboratory line to be infected with a wAu-like Wolbachia. Moreover, we observed that all Drosophila willistoni samples tested with the VNTR-141 marker harbor the same Wolbachia variant, wWil, either in populations from the South or the North of Brazil. Horizontal transfer events involving species of Old World immigrants and Neotropical species of the willistoni subgroup are discussed.

BuT2 Is a Member of the Third Major Group of hAT Transposons and Is Involved in Horizontal Transfer Events in the Genus Drosophila

The hAT superfamily comprises a large and diverse array of DNA transposons found in all supergroups of eukaryotes. Here we characterized the Drosophila buzzatii BuT2 element and found that it harbors a five-exon gene encoding a 643-aa putatively functional transposase. A phylogeny built with 85 hAT transposases yielded, in addition to the two major groups already described, Ac and Buster, a third one comprising 20 sequences that includes BuT2, Tip100, hAT-4_BM, and RP-hAT1. This third group is here named Tip. In addition, we studied the phylogenetic distribution and evolution of BuT2 by in silico searches and molecular approaches. Our data revealed BuT2 was, most often, vertically transmitted during the evolution of genus Drosophila being lost independently in several species. Nevertheless, we propose the occurrence of three horizontal transfer events to explain its distribution and conservation among species. Another aspect of BuT2 evolution and life cycle is the presence of short related sequences, which contain similar 5′ and 3′ regions, including the terminal inverted repeats. These sequences that can be considered as miniature inverted repeat transposable elements probably originated by internal deletion of complete copies and show evidences of recent mobilization.

A low-cost apparatus for transforming Drosophila and detecting green fluorescent protein (GFP) genetic markers

We describe the transformation of white mutant Drosophila simulans with a piggyBac transposon vector and a green fluorescent marker (GFP) and show how to construct inexpensive micro-manipulation and epifluorescence equipment for use in transposon-mediated germ-line transformation. Although the number of G0 adult flies (16) obtained in relation to the number of injected eggs was very low (12.5%) it was comparable to the proportion described by other authors and can be considered as a good rate of transformation.

Effects of Irradiation Dose on Sterility Induction and Quality Parameters of Drosophila suzukii (Diptera: Drosophilidae)

Abstract Drosophilasuzukii(Matsumura, 1931) (Diptera: Drosophilidae) is a widely distributed pest of soft-skinned and stone fruits that is controlled mainly with pesticides. An alternative to the chemical control is the sterile insect technique (SIT), an ecologically friendly method of pest management that could be used against D. suzukii.The objective of the present study was to evaluate the effects of gamma radiation on reproductive sterility, ovarian morphometry, and quality parameters of D. suzukii. Full female sterility was achieved at 75 Gy, while an adequate level of male sterility (99.67%) was obtained at 200 Gy.The ovarian size showed an exponential decay in function of irradiation dose increase.There was no significant influence of irradiation dose on the quality parameters evaluated. Our data suggest that gamma radiation can be recommended to be used in an SIT program for D. suzukii.

Total evidence phylogenetic analysis and reclassification ofEuschistusDallas within Carpocorini (Hemiptera: Pentatomidae: Pentatominae): Total-evidence phylogeny ofEuschistus

Robust phylogenetic hypotheses have become key for studies addressing the evolutionary biology and ecology of various groups of organisms. In the species‐rich heteropteran superfamily Pentatomoidea, phylogenies at lower taxonomic levels are still scarce and mostly employ exclusively morphological data. In this study, we conducted a total evidence phylogeny focusing on the tribe Carpocorini (Pentatomidae), using morphological data and four DNA markers (COI, Cytb, 16S and 28S rDNA; ∼2330 bp; 32 taxa) in order to investigate the relationships within Euschistus Dallas, one of the most speciose pentatomid genera, and between Euschistus and related genera. Our hypotheses generated by maximum likelihood and Bayesian inference show that the current taxonomic composition and classification of Euschistus and allied genera are in need of revision. Euschistus was recovered as nonmonophyletic, with the subgenera forming four independent lineages: Euschistus (Euschistus) and Euschistus (Lycipta) Stål are sister groups; Euschistus (Euschistomorphus) Jensen‐Haarup is more closely related to Dichelops Spinola and Agroecus Dallas; and Mitripus Rolston is divided into two clades closely related to Sibaria Stål and Ladeaschistus Rolston. We chose not to change the classification of E. (Euschistomorphus) until further data become available, and propose to split Euschistus into three genera with the exclusion of Euschistus (Mitripus) and all of its species. Here we elevate Mitripus to genus rank to include M. acutus comb.n., M. convergens comb.n. and M. legionarius comb.n., and propose Adustonotus Bianchi gen.n. to include A. anticus comb.n., A. latus comb.n., A. tauricornis comb.n., A. grandis comb.n., A. hansi comb.n., A. paranticus comb.n., A. irroratus comb.n. and A. saramagoi comb.n. We also provide identification keys to the genera Adustonotus gen.n., Ladeaschistus, Mitripus n. rank and Sibaria, here defined as the Mitripus genus group, and to the species of Mitripus and Adustonotus gen.n. Our results provide insights into the current status of the classification of the Pentatomidae, suggesting the need for phylogenetic analyses at different taxonomic levels within stink bugs.

Radiation effects on Drosophila suzukii (Diptera: Drosophilidae) reproductive behaviour

Female remating is a widespread behaviour, reported in several insect species. This behaviour can affect the efficiency of sterile insect technique (SIT); however, little is known about the postcopulatory behaviour of some pest species considered as candidates to be controlled by this technique, such as Drosophila suzukii (Matsumura, 1931) (Diptera: Drosophilidae). In this study, we investigated the effects of male and female sterilization on mating and remating behaviour of D. suzukii. First, we tested the occurrence of multiple mating in different combinations between sterile and fertile males and females. Then, we tested the effects of male and female sterility on female propensity to mate and remate. We found an overall low remating rate by D. suzukii females. Male sterility did not influence mating and remating likelihood; however, copula duration of sterile males was shorter compared to fertile males. On the other hand, sterile females were less likely to mate. Our findings encourage further research regarding the use of SIT to control D. suzukii.