Unveiling the Mycodrosophila projectans (Diptera, Drosophilidae) species complex: Insights into the evolution of three Neotropical cryptic and syntopic species.

The Zygothrica genus group has been shown to be speciose, with a high number of cryptic species. DNA barcoding approaches have been a valuable tool to uncover cryptic diversity in this lineage, as recently suggested for the Neotropical Mycodrosophila projectans complex, which seems to comprise at least three different species. The aim of this study was to confirm the subdivision of the M. projectans complex while shedding some light on the patterns and processes related to its diversification. In this sense, the use of single and multi-locus datasets under phylogenetic, distance, coalescence, and diagnostic nucleotide approaches confirmed the presence of at least three species under the general morphotype previously described as M. projectans. Only a few subtle morphological differences were found for the three species in terms of aedeagus morphology and abdominal color patterns. Ecologically, sympatry and syntopy seem to be recurrent for these three cryptic species, which present widely overlapping niches, implying niche conservatism. This morphological and ecological similarity has persisted though cladogenesis within the complex, which dates back to the Miocene, providing an interesting example of morphological conservation despite ancient divergence. These results, in addition to contrasting patterns of past demographic fluctuations, allowed us to hypothesize patterns of allopatric or parapatric diversification with secondary contact in Southern Brazil. Nevertheless, genetic diversity was generally high within species, suggesting that migration may encompass an adaptive response to the restrictions imposed by the ephemerality of resources.

Phylogenetic relationships between fungus-associated Neotropical species of the genera Hirtodrosophila, Mycodrosophila and Zygothrica (Diptera, Drosophilidae), with insights into the evolution of breeding sites usage.

The Neotropical region harbors an astonishing diversity of species, but still encompasses the least studied biogeographic region of the world. These properties apply for different taxonomic groups, and can be exemplified by drosophilids. In fact, high levels of cryptic diversity have recently been discovered for Neotropical species of the Zygothrica genus group, but relationships among these species, or them and other Drosophilidae species still remains to be addressed. Therefore, the aim of this study was to evaluate the phylogenetic relationships between fungus-associated Neotropical species of the genera Hirtodrosophila, Mycodrosophila and Zygothrica, which together with Paramycodrosophila and Paraliodrosophila compose the Zygothrica genus group. For this, fragments of the mitochondrial cytochrome oxidase subunits I (COI) and II (COII) genes, and the nuclear alpha methyldopa (Amd) and dopa decarboxylase (Ddc) genes were newly characterized for 43 Neotropical specimens of fungus-associated drosophilids, and analyzed in the context of 51 additional Drosophilinae sequences plus one Steganinae outgroup. Based on the resulting phylogeny, the evolution of breeding sites usage was also evaluated through ancestral character reconstructions. Our results revealed the Zygothrica genus group as a monophyletic lineage of Drosophila that branches after the subgenera Sophophora and Drosophila. Within this lineage, Mycodrosophila species seem to encompass the early offshoot, followed by a grade of Hirtodrosophila species, with derived branches mostly occupied by representatives of Zygothrica. This genus, in particular, was subdivided into five major clades, two of which include species of Hirtodrosophila, whose generic status needs to be reevatuated. According to our results, the use of fungi as breeding sites encompasses a symplesiomorphy for the Zygothrica genus group, since one of the recovered clades is currently specialized in using flowers as breeding sites whereas a sole species presents a reversal to the use of fruits of a plant of Gentianales. So, in general, this study supports the paraphyly of Drosophila in relation to fungus-associated Neotropical species of Drosophilidae, providing the first molecular insights into the phylogenetic patterns related to the evolution of this diverse group of species and some of its characteristic traits.

The DNA barcoding and the caveats with respect to its application to some species of Palaemonidae (Crustacea, Decapoda).

DNA-barcoding has recently attracted considerable attention due to its potential utility in aiding in species identification and discovery through the use of a short standardized sequence of mitochondrial DNA. Nevertheless, despite the fact that this technology has been proven a useful tool in several animal taxa, it also demonstrated limitations that may hinder correct application. Thus, its validity needs to be empirically evaluated in each taxonomic category before forward implementation. As the use of DNA barcoding within Palaemonidae may be of special interest, given its great interspecific morphological conservatism associated with considerable intraspecific morphological variation, we analyze here the potential of this technology in distinguishing and recovering some taxonomic boundaries within this family. We asked whether two GenBank-retrieved sets of COI sequences encompassing the conventional Barcode and Jerry-Pat regions possess the desired properties of reciprocal monophyly among species, and existence of a barcoding gap between intra- and interspecific variations, after performing a careful analysis of numt (nuclear mitochondrial DNA) contamination. These analyses revealed nine non-monophyletic species, with some cases of divergent intraspecific sequences, contrasted with interspecific similarity attained in others. Moreover, we were unable to identify any barcoding gap between intraspecific and interspecific divergences within Palaemonidae, although a threshold of 0.18 substitutions per site would differentiate intraspecific and congeneric divergences in 95% of the cases for the barcoding region. A fraction of the overlap could be certainly attributed to artifacts related to poor taxonomy, but even from this perspective DNA barcoding studies may help to uncover previously disregarded taxonomic and evolutionary issues.