Fig 1.
Median joining haplotype network obtained from COI gene sequences of Philaenus spumarius collected from different regions of Italy.
(A) Haplotype frequency by geographic region. (B) Haplotype frequency by Wolbachia infection. Circle sizes are proportional to haplotype frequency. Colors correspond to the region of origin of the haplotype (A) and the Wolbachia infection status (B). Small black dot vertices represent missing or unsampled haplotypes. Diamonds on branches represent the number of mutations. No diamond on branches means one mutation.
Table 1.
Wolbachia infection in Italian populations of Philaenus spumarius.
Table 2.
PCR detection of Wolbachia in the head of infected individuals of Philaenus spumarius.
Percentage of infected heads calculated on the number of infected individuals.
Table 3.
Wolbachia sequence types (ST) in Philaenus spumarius and their association with host population and haplotype.
New STs and allelic variants in the MLST database are in italic.
Fig 2.
Frequency of mitochondrial lineages of Philaenus spumarius (histograms) and Wolbachia infection rate (cakes) in the sampled regions of Italy.
The percentage of infected individuals is the slice of cake colored with checkered texture. Similarly, regions with populations of P. spumarius infected by Wolbachia are colored with checkered texture on the map. The blue big cakes represent the Wolbachia infection rate calculated on the total number of individuals sampled in each region. The small cakes represent the Wolbachia infection rate calculated on the individuals of each mitochondrial lineage. The map of Italy by Sinigagl (https://commons.wikimedia.org/wiki/File:Italy_template_blank.png) is licensed under CC BY-SA 3.0; it is similar but not identical (it differs for the colored regions) to the original map and is therefore for illustrative purposes only.
Table 4.
Wolbachia infection of mitochondrial lineages of Philaenus spumarius.
Fig 3.
Bayesian phylogenetic tree for wsp gene sequences of Wolbachia.
Strains of Wolbachia found in Philaenus spumarius and sequenced in this work are shown in bold red. Reference Wolbachia strains were imported from the MLST database. Wolbachia strains are indicated with the name of the host species followed by the number of the wsp allelic variant. Based on the information reported in the MLST database for the reference strains, the letters on the right indicates the Wolbachia supergroups.
Fig 4.
Bayesian phylogenetic tree for concatenated MLST gene sequences of Wolbachia.
Wolbachia STs found in Philaenus spumarius and sequenced in this work are shown in bold red. Reference Wolbachia STs were imported from the MLST database and are indicated on the tree with the name of the host species and the number of the ST. Wolbachia strains of P. spumarius by [53] are reported with the original strain number (ST number not available in the MLST database) and those from Italy are in bold. Based on the information reported in the MLST database for the reference STs, the letters on the right indicates the Wolbachia supergroups.
Fig 5.
Neighbour-net phylogenetic network based on concatenated MLST gene sequences of Wolbachia.
Wolbachia STs found in Philaenus spumarius and sequenced in this work are shown in red (IT means Italy). Wolbachia strains of P. spumarius by [53] are shown in green with the original strain number preceded by the acronym of the Country of origin. Reference Wolbachia STs were imported from the MLST database and are indicated on the tree with the name of the host species and the number of the ST. The capital letters indicates the Wolbachia supergroups.
Fig 6.
Fluorescence in-situ hybridization of Philaenus spumarius.
Ovarioles (A and B) and salivary glands (C and D) stained by a Wolbachia and a Sulcia specific probes. (A) Distribution of Wolbachia bacteria (bright red) within telotrophic ovarioles and Sulcia primary symbionts (green) penetrated into developing oocytes. (B) Wolbachia symbionts (bright red, arrow) and the ball-shaped mass of Sulcia symbionts (green) localized at the posterior pole of a developed oocyte; autofluorescence of ooplasm is visible. (C and D) Wolbachia bacteria (bright red) distributed within an upper (C) and a ventral (D) salivary gland. DAPI-stained nuclei are blue. Bars, 100 μm.