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Drosophila lehrmanae sp. nov. (Fig. 10) Holotype. Male. FRENCH GUIANA: Camp Nouragues, Inselberg, coordinates: 4°05’ N - 52°41’W, June 2015, W.J. Miller and A. Hua-Van, MNHN - Museum National d’Histoire Naturelle in Paris / France. (Fig. 10 A-C) Paratypes. 9 males and 10 females FRENCH GUIANA: Camp Nouragues, Inselberg, coordinates: 4°05’ N - 52°41’W, June 2015, W.J. Miller and A. Hua-Van, MNHN- Museum National d’Histoire Naturelle in Paris / France (4 males and 5 females); MZSP- Museum of Zoology of University of São Paulo State, Brazil (5 males and 5 females) (Fig. 10 D-F). Etymology. The name D. lehrmanae sp. nov. was chosen in honor of the Distinguished Professor Emerita Lee Ehrman, a student of Theodosius Dobzhansky and outstanding drosophilist at the State University of New York at Purchase, NY, USA, who significantly contributed to our understanding of evolutionary and behavioral biology over the last 60 years of her academic career (Kim 2005). Diagnosis. On closer examination, D. lehrmanae sp. nov. differs from D. sturtevanti in the following diagnostic characters: The body coloration (mesonotum, pleura, scutellum and abdomen) is lighter; the antennae are dark brown; females: the spot on the sixth tergite is smaller and its format is more like a circle than a half-moon; the spermatheca presents two collars instead of one; males: the aedeagus presents a relatively shorter apex. Description. Male. Head: Arista with 4-5 dorsal and 2 ventral branches, with bifurcated termination. Antennae dark brown, the proximal segment (pedicel) presents the light brown margins. Front velvety brown with area around ocellar triangle and around the orbital bristles light brown. Ocellar triangle and region of orbital setae darker. Anterior orbital about 1/3 posterior and middle orbital setae. Carina light brown, not sulcate, slightly prominent, and with small bristles below. Palpi light brown, pollinose, with bristles on the ventral surface and one very prominent on the end. Two prominent oral bristles. Greatest width of the cheeks about 1/6 to 1/8 greatest diameter of eye. Eyes dark red with a short dark pile. Thorax: mesonotum light brown, it presents two darker uninterrupted longitudinal stripes in the central region, which extends from the anterior portion of the mesonotum and continues until the end of scutellum. It also presents a darker longitudinal stripe on each side, interrupted approximately in the first anterior quartile of the mesonotum and it does not continue until the scutellum. Acrostichal hairs in 6 rows. No prescutellars. Anterior scutellars parallel. Halteres light color. Scutellum light brown with indistinct continuations of the darker stripes of the mesonotum. Pleurae brown. Anterior sternopleural 1/3 posterior and much thinner. Coxae, femora, tibiae and tarsos yellowish brown. Abdomen: light brown with a longitudinal dark stripe on the inferior margins of each tergite. Male terminalia: The dorso-ventral region of the epandrium is angular. The cercus is U-shaped, covered with cercal bristles. The surstyli are large and concave and they are connected by a small subepandrial sclerite (decasternum). Wings: Wings hyalines. Costal index 1.8—2.0. Body length: 2.4 mm. Female: The body features are the same as in males, but the sixth tergite is lighter and it presents a small opaque circular spot on each side. Approximately eight days after the puparia hatch, the adult female shows a clear contour in the opaque circular area of the sixth tergite. Costal index 2.0—2.2. Body length: 3.1 mm. The main morphological characteristics of the species included in the sturtevanti subgroup described in works in the literature were compared with those of new species Drosophila lehrmanae sp. nov. (Table 9). The genetic markers also were also able to distinguish the species, for COI5’ there were 23, 26 and 23 mutation, for COI3’there were 28, 26 and 25, COII 20, 19 and 18 and for ND4 15, 13 and 9 comparing with D. dacunhai, D. milleri and D. sturtevanti, respectively. The mutation and its sites are seen in Table 10.

Review history. (DOCX 31 kb)

Additional figures. Figure S1. Overlap in Drosophila gene content between the transcriptomes of the three semispecies. Figure S2. Principal component analysis (a) and heatmap (b) of expression data of the AM semispecies. Figure S3. Principal component analysis (a) and heatmap (b) of expression data of the CA semispecies. Figure S4. Number of genes differentially expressed in one or multiple conditions of the AM semispecies. Figure S5. Number of genes differentially expressed in one or multiple conditions of the CA semispecies. Figure S6. First and second principal components in the PCA of female abdomen samples of all semispecies. Figure S7. Second and third principal components in the PCA of head samples of all semispecies mapped to the OR transcriptome. Figure S8. Principal component analysis of GFR head samples of all semispecies mapped to the OR transcriptome based on the same genes used in the WT head PCAs (Figs. 5c, 2d). Figure S9. Principal component analysis of abdomen samples of all semispecies mapped to the OR transcriptome. (PDF 2115 kb)

Effect of the time point of gamma-irradiation on CHC profiles of individually reared 10Â day old adult G. m. morsitans males. Discriminant analysis based on log-ratio transformed relative amounts across irradiation time points (early and late pupal development and as young adults). (PDF 1354Â kb)

Comparison of CHC profiles of untreated (Ctr) and tetracycline-treated (Tet) (a) female and (b) male tsetse flies (G. m. morsitans) across the three different experiments. (PDF 1363Â kb)

Effect of gamma-irradiation dose and time point on CHC profiles of 10Â day old individually reared adult G. m. morsitans males. Discriminant analysis based on log-ratio transformed relative amounts across all treatment groups (time points and irradiation doses). (PDF 1353Â kb)

Table S1. Sequences and positions of the 48 oligos within the Wolbachia 16S and 23S ribosomal RNA used for StellarisÂŽ rRNA-FISH. (PDF 90 kb)