Berberigetta dimelodica, Costa, Gonçalo João, Nunes, Vera L., Marabuto, Eduardo, Mendes, Raquel, Laurentino, Telma G., Quartau, José Alberto, Paulo, Octávio S. & Simões, Paula Cristina, 2017

Costa, Gonçalo João, Nunes, Vera L., Marabuto, Eduardo, Mendes, Raquel, Laurentino, Telma G., Quartau, José Alberto, Paulo, Octávio S. & Simões, Paula Cristina, 2017, Morphology, songs and genetics identify two new cicada species from Morocco: Tettigettalna afroamissa sp. nov. and Berberigetta dimelodica gen. nov. & sp. nov. (Hemiptera: Cicadettini), Zootaxa 4237 (3), pp. 517-544 : 530-533

Berberigetta dimelodica sp. nov. Costa, Nunes, Marabuto, Mendes & Simões

Material examined Paratypical series consists of a total of 14 specimens (13 males and one female). Designated holotype is SP19_3795 (♂), and female paratype is SP19_3787 (♀). See Table 2 for additional information on paratypical series, specimen IDs, collection sites and GPS data. See Figure 6 for images on male holotype, female paratype (see supplementary image, S6 for live specimens) and details of the male genitalia.

Male morphology

Head Supra-antennal plate produced into a pointed lobe; Supra-antennal plate nearly meeting the eye. Postclypeus subquadrate to round in front view; Postclypeus transversely grooved towards distal ends. Rostrum brown, reaching the center of mid-trochanters when in resting position. Antennae brown, 7-segmented. Postclypeus dark brown, with apical yellowish-brown spot, grooves light-brown or yellowish; Anteclypeus yellowish with a brown central spot. Gena and lorum brown to light-brown covered with white long pilosity. Supra-antennal plates light brown distally near the eye, becoming dark-brown towards midline. Three red ocelli. Eyes light-brown. Dorsal surface of head dark-brown, supraocular border brown, with yellowish stripe on epicranial suture.

Thorax Pronotal collar broad, slightly greater than eye width; Pronotal lateral development ampliate, sloping in lateral view, evenly rounded in dorsal view. Pronotal mid-lateral tooth absent. Scutellum wider than long. Epimeral lobe not reaching operculum. Metanotum partly visible at dorsal midline, not expanded over tymbals. Pronotum brown with a dark-brown stripe along dorsal midline, ending posteriorly in dark-brown spot. Mesonotum with two yellowish fasciae bordering between parapsidal suture and submedian sigillae prolonging to anterior arms of scutellum; Mesonotal lateral dorsal margins yellowish. Central area of scutellum brown with yellowish arms. Metanotum yellowish, brown at dorsal midline.

Legs Profemur with a large primary erect spine plus two smaller secondary spines dark-brown/ brownish in colour, some individuals with a much smaller fourth spine. Meracanthus triangular. Tarsal formula 3-3-3. General brown to yellowish in colour. Metatibiae with four long fine reddish spurs on inner side and two smaller reddish spurs on outer side. Coxae yellowish, with a central dark-brown stripe, becoming gradually browner and less yellowish towards metacoxae. Trochanters brown. Meso and metafemurs yellowish with dark-brown to brownish stripes. Tarsi and tibia light-brown.

Wings Forewing with eight apical and four subapical cells. Ulnar cell 3 angled to radial cell. Costal vein parallel-sided to node. Pterostigma present becoming darker towards distal end. CuA weakly bowed. M and CuA meeting at basal cell with stems completely fused. RA1 slightly diverging from subcostal at subapical region before crossvein. C and R+Sc close together. CuP and 1A non-fused at their bases. Forewing outer margin developed for its total length. Membrane hyaline. Hindwing vein 2A with an infuscation running alongside total length of vein. First cubital cell width at distal end much greater than second cubital cell. Anal lobe broad, with vein 3A bowed at distal end. Larger forewing proximal veins yellowish with smaller apical veins brown, same vein colour pattern for hindwing. Costal vein yellowish. Basal membrane and plaga yellowish.

Opercula More or less confluent with distal margin of tympanal cavity, well developed towards abdominal midline with sharply rounded apices facing midline. General opercula colour yellowish becoming brown at the base. Meracanthus following the same colour pattern as opercula.

Tymbals Tymbal covers absent. Four to five ribs, broadening apically, three of which arising from anterior proximal part of a large basal dome covering over half total length of tymbal. First anterior rib is slender, with a break at about a third of its length. Fourth rib arising from anterior distal side of basal dome more or less evident amongst individuals. Some specimens present a fifth less defined rib arising from posterior distal end of the basal dome, transversal to fourth rib and converging in a sharp end. Tymbal ribs and basal dome brownish-grey; tymbal plate light-grey.

Abdomen Tergites T2 and T3 much enlarged accounting for about a third of total abdominal length. StVIII greater in length than StVII. T1 and T2 dark-brown; T4–7 dark-brown on dorsal midline, sides red and covered in fine silvery pubescence; T8 dark-brown on dorsal midline, sides yellowish. Sternite I brown; StII yellowish with a brown patch on elevated central area; StIII–VIII yellowish. Epipleurites yellowish.

Genitalia ( Figure 6 C to 6F) Pygophore distal shoulder not developed; Pygophore inner tooth absent; Upper lobe present, small and rounded, distant from dorsal beak; Basal lobe small to moderately developed ending in a sharp, rounded tip, in lateral view. Dorsal beak well developed, sharp and part of chitinized pygophore. Ventrobasal pocket absent. Claspers small-medium sized, hooked slightly outwards on distal end, rounded tip. Uncus duck-bill shaped, small and flat, not dominant and retractable within pygophore; Uncus lateral lobes absent. Aedeagal basal plate, undulated in lateral view, weakly depressed on dorsal midline; Basal plate apically broad, flat and rounded in ventral view, with a medial small sharp-tipped lobe on both sides, followed by a tubelike constriction leading to theca, gradually narrowing, slight medial lateral depression; Basal plate bearing a ripple-like pattern in dorsal view. Basal portion directed forwards away from thecal shaft; Ventral rib not apparent; Basal plate completely fused to theca without mobility. Theca very long and J-shaped in lateral view. Thecal pseudoparamers lateral of theca, dorsally fused until two thirds of theca length, very flat, as long as endotheca, ending on an upward pointed, sharp tip; Ventral support absent. Pygophore dorsal surface lightbrown to yellow. Claspers dark-brown. Uncus brown.

Female morphology Only one female known so far (see supplementary image, S6 for the live specimen). Generally lighter in colour than male. Postclypeus yellowish with brown grooves, genae and lora light brown; Legs generally light brown; Dorsal surface of head light-brown with brown patterns; thorax and scutellum lightbrown. Abdomen light brown laterally, with a lighter brown on dorsal midline.

Body measurements for 13 males of B. dimelodica Total length: 16.99 ± 0.78 mm; Pronotal length: 1.74 ± 0.15 mm; Mesonotal length: 2.67 ± 0.11 mm; Forewing length: 13.39 ± 0.54 mm; M+CuA length: 1.21 ± 0.18 mm. Female and additional body measurements can be found on Table 3.

Bioacoustics The calling song here described is based on the analysis of recordings of three males singing at T=39–40 °C. A typical phrase is structured into four sequential parts ( Figure 7 ): A, a single echeme; B, a series of 16 ± 2.60 echemes (10–21, n=52) in rapid succession; C, a group of 8 ± 3.68 echemes (5–18, n= 53) ending on D, a single, long echeme. In 21.15% and 9.61% of the phrases part A and part D are missing, respectively.

Calling song frequency-based analysis revealed an interesting frequency modulation in part B. Peak frequency for parts A, C and D is 13.88 ± 0.79 kHz, with maximum frequency of 20.65 ± 0.54 kHz. During part B there is an abrupt reduction of the frequency with a peak frequency of 7.91 ± 1.62 kHz, yet, maintaining the maximum frequency at 21.62 ± 1.11 kHz.

For additional time and frequency variables consult Table 6. Note that, due to frequency modulation in part B, it was separated from parts A, C and D in our analysis.

B. dimelodica Phrase Part A Part B

Time variables Mean±SD Min–Max n Mean±SD Min–Max n Mean±SD Min–Max n

Duration (ms) 2218 ± 559 1357–3448 52 30 ± 10 15–56 47 335 ± 52 212–411 52

Echeme duration (ms) - - - Same as above 2.14 ± 1.06 0.8–7 849

Echeme rate (echeme.s -1) - - - - - - 49.16 ± 6.08 36.08–72.67 52

Interval (ms) 259 ± 82 195–614 49 - - - 19.55 ± 5.31 2.8–55 797

Part C Part D

Time variables Mean±SD Min–Max n Mean±SD Min–Max n

Duration (ms) 1364 ± 679 632–2992 53 252. 29 ± 79.23 97–430 41

Echeme duration (ms) 49.2 ±20.6 5–253 487 Same as above

Echeme rate (echeme.s -1) 7.10 ± 1.04 3.34–10.32 53 - - -

Interval (ms) 108.83 ± 22.24 34–260 435 - - -

continued.

Frequency variables Peak frequency Min frequency Max frequency Bandwidth

Part ACD Mean ± SD 13.88 ± 0.79 4.65 ± 0.96 20.65 ± 0.54 15.94 ± 1.31 Min–Max 11.50–15.50 1.96–6.00 18.70–22.40 12.80–19.78

Part B Mean ± SD 7.91 ± 1.62 4.39 ± 1.01 21.62 ± 1.11 17.14 ± 1.59 Min–Max 5.60–16.50 0.30–5.80 13.92–23.40 9.04–22.80

Frequency variables Quartile 25 Quartile 50 Quartile 75 Quartile (75%–25%)

Part ACD Mean ± SD 11.91 ± 0.22 13.48 ± 0.27 14.89 ± 0.32 2.98 ± 0.26

Min–Max 10.70–12.50 12.28–14.40 13.40–15.80 1.87–4.10

Part B Mean ± SD 7.54 ± 0.61 9.57 ± 0.79 11.61 ± 1.26 4.07 ± 0.93

Min–Max 6.30–12.00 7.50–13.50 9.70–17.50 2.70–8.20 DNA barcoding Four haplotypes were recovered among the COI sequences of nine males of B. dimelodica sp. nov., with a nucleotide diversity of π= 0.0164. Sequences were clustered into two well supported sister clades ( Figure 4 ) diverging by 2.9 % (K2P distance). These clades are, according to our currently knowledge, geographically segregated. Among the 18 segregating sites observed, 16 are fixed for each clade, being two of them non-synonymous mutations. Mean interspecific genetic distances for B. dimelodica are presented in Table 5 . The new species is clearly distinguishable within the Cicadettini ( Tettigettalna , Tettigettacula , Tympanistalna , Euryphara and Hilaphura ), with mean pairwise genetic distances>10%. The COI fragment is therefore apparently proficient for DNA barcoding of B. dimelodica , though the genetic structure reported here must be taken into account.

Distribution ( Figure 1) Morocco, in the northern parts of Middle Atlas Mountains, near Taza and along the eastern Rif mountains (Al Hoceima), eastward to Berkane (Beni-Snassen Mountains), as the extreme western foot of the Tellian Atlas Mountains. On biogeographical grounds it is possible that this species is also in western Algeria.

Habitat ( Figure 5) Open scrubland or light xerothermophilous woodland dominated by holm-oak ( Quercus rotundifolia ) in the northern Middle Atlas or mixed pinewoods of Pinus halepensis and Tetraclinis articulata with a rich understory of Pistacia lentiscus , Chamaerops humilis , Rosmarinus officinalis and Stipa spp. Males sing mainly perched on these shrubs, and sometimes on the lower branches of trees (<3 m height).

Etymology Specific epithet dimelodica arises from the dual sound production during the calling song of this species, meaning “two melodies”. It consists of two distinct sound patterns, with the second part severely downshifted in frequency and resembling a human-produced unvoiced linguolabial trill, often referred as “Blowing a raspberry”.

Figures

FIGURE 4. Bayesian inference phylogenetic tree of Cytochrome C oxidase subunit I mitochondrial DNA of T. afroamissa and B. dimelodica with other previous published taxa. Posterior probabilities are shown next to branch nodes. TET stands for Tettigettacula — Euryphara — Tympanistalna clade. Scale bar represents the number of estimated changes per branch length. C. barbara (Cba 203) and C. orni (Cor 298) were set as an outgroup. T. afroamissa and B. dimelodica taxa IDs are detailed on Table 2. Additional taxa details are included on supplementary information Table S 1. Root was truncated with double dash totalling 0.6 changes per branch length.

FIGURE 6. Body and male genitalia morphology of Berberigetta dimelodica. A—Designated male holotype of B. dimelodica. Scale bar equals 10 mm; B—Designated female paratype of B. dimelodica. Scale bar equals 10 mm; C, D—Male paratypes’ pygophore overview in posterior and lateral views, respectively. Scale bars equal 500 µm. E, F—Aedeagus in upper and lateral views, respectively. Scale bars equal 200 µm. Pygophore and aedeagus photos were taken of material preserved in Kaiser gelatin. Note that the tip of the left pseudoparamer is broken.

FIGURE 7. Berberigetta dimelodica calling song profile. Mean frequency spectrum (1), oscillogram (2) and spectrogram (3). Letters A, B, C and D refer to the structural divisions found in a typical phrase. Individualized analysis of part B and parts C, D and A (sequentially) are displayed in the bottom graphs. Calling song recorded on Middle Atlas, Afouzar at 38 – 40 ° C.