Molecular phylogeny of the Mediterranean species ofPhilaenus(Hemiptera: Auchenorrhyncha: Aphrophoridae) using mitochondrial and nuclear DNA sequences

Abstract

The phylogenies of all eight European species of Philaenus were estimated from cytochrome oxidase subunit I, cytochrome B and internal transcribed spacer 2 (ITS2) fragments of DNA using phylogenetic reconstruction methods: maximum parsimony (MP), maximum likelihood (ML) and Bayesian inference (BI) analyses. Based on the topologies of all obtained phylogenetic trees, the monophyly of Philaenus is well supported, being congruent with morphological, ecological and chromosomal data. Three phylogenetic lineages were distinguished in the mitochondrial and combined (mtDNA with ITS2) trees. The first lineage is represented by only one species, Philaenus maghresignus, which inhabits Maghreb and southern Spain. Clade A includes three species: P. tarifa (Southern Iberia), P. italosignus (Sicily and Southern Italy) and P. signatus (the Balkans and Middle East). In clade B two subclades were recognized: B1 represented by P. loukasi (Southern Balkans) and P. arslani (Middle East), and B2 comprising P. spumarus (the most widespread Palaearctic species) and P. tesselatus (from Southern Iberia and Maghreb). These clades were also retrieved in trees reconstructed from nuclear sequences. However, four species (P. maghresignus, P. tarifa, P. italosignus and P. signatus) showed unresolved polytomy at the base of the nuclear tree. Clade A together with P. maghresignus clustered with the 'signatus' group defined from morphology, and clade B with the 'spumarius' group; these might be considered separate subgenera. Genetic distances in mitochondrial DNA between ingroup species ranged from 14.0% between P. signatus and P. spumarius to 2.4% between P. tesselatus and P. spumarius. By contrast, genetic divergence of ITS2 between ingroup species was very low, at most 2.1%. The divergence of Philaenus species is estimated to have occcurred between 7.9 and 0.6 Ma. Possibly three main speciation events occurred: the first at the Miocene/Pliocene boundary (c. 5.5 Ma) for deeper splits; the second between 4.2 and 2.5 Ma in the Pliocene, when pairs of more closely related species diverged; and the most recent during the Pleistocene glaciations, when the separation of P. tesselatus and P. spumarius took place. The species status of all Philaenus species is confirmed except for P. tesselatus.

The Mediterranean: Area of origin of polymorphism and speciation in the spittlebug Philaenus (Hemiptera, Aphrophoridae)

“…Therefore, during the last four years molecular and cytogenetic studies of new material of the eight species was analyzed. The results of these new studies reinforced the new systematic and ecological behaviour of each of the eight new taxa (Loukas & Drosopoulos 1993;Maryańska-Nadachowska 2008a). Based on the new data another approach for explaining the phenomenon of polymorphism in each species of this interesting spittlebug genus is reported here.…”

“…Recent taxonomic (Drosopoulos & Asche 1991;Abdul-Nour & Lahoud 1995;Drosopoulos & Remane 2000), ecological (Drosopoulos 2003), genetic (Loukas & Drosopoulos 1993), molecular (Maryańska-Nadachowska et al 2008a) and caryological (Maryańska-Nadachowska et al 2008b) studies in the whole Mediterranean revealed that dorsal colour polymorphism in the spittlebug Philaenus spumarius is more complicated than reported earlier (Halkka 1962;Thompson 1973;Stewart & Lees 1988). These studies warranted that in addition to two then recognized species, the Holarctic P. spumarius and the Mediterranean P. signatus six more species occur in the Mediterranean.…”

High genetic diversity in southwest Asian populations ofPhilaenus spumarius(Hemiptera: Auchenorrhyncha)

“…DNA isolation, amplification, sequencing, and analyses were executed exactly as in the previous works of Maryańska-Nadachowska et al (2010. We amplified two markers: mitochondrial cytochrome B gene (CytB) and internal transcribed spacer 2 (ITS2) of nuclear ribosomal DNA -markers which were used for previous studies on the phylogeny of the Philaenus species (Maryańska-Nadachowska et al, 2010) and on the phylogeography of P. spumarius (Maryańska-Nadachowska et al, 2012;Lis et al, 2014).…”

“…The Palaearctic genus Philaenus Stål, 1864 is one of the most intensively studied in Hemiptera, previously mainly due to an extremely high level of colour polymorphism of individuals (e.g., references in Halkka & Halkka, 1990;Stewart & Lees, 1996;Drosopoulos, 2003), and lately due to interesting findings about morphological, ecological, karyological, and molecular diversity (Kuznetsova, Maryańska-Nadachowska, & Nokkala, 2003;Maryańska-Nadachowska, Drosopoulos, Lachowska, Kajtoch, & Kuznetsova, 2010;Maryańska-Nadachowska, Kajtoch, & Lachowska, 2012;Seabra et al, 2010), which allowed the identification of seven new species within the last three decades. (Lis, Maryańska-Nadachowska, & Kajtoch, 2015).…”

Molecular barcoding for central-eastern European Crioceris leaf-beetles (Coleoptera: Chrysomelidae)

“…The utility of the COI mitochondrial gene as a DNA barcoding system for the identification or verification of the taxonomic status of species has been tested in many animal groups and has shown promising results in some cases (e.g., for leaf-beetles and weevils: [49][50][51][52]) and bugs [53,54]), but also some constraints in others (e.g., [25,55,56]). Similar research on the application of DNA barcoding for testing taxonomic and phylogenetic species concepts of closely related species of leafbeetles were performed e.g.…”