The genus Tibicina Kolenati, 1857 in Morocco (Hemiptera: Cicadidae: Tibicininae): taxonomic assessment from integrative research Author Puissant, Stephane 0000-0003-4179-3636 Muséum national d’Histoire naturelle, Département Systématique et Evolution, UMR CNRS 7205 OSEB, CP 50, 45 rue Buffon, F - 75005 Paris and Muséum d’Histoire naturelle de Dijon - Jardin de l’Arquebuse, 14 rue Jehan de Marville, F - 21000 Dijon, France. 10 chemin des Châtaigniers, F - 74150 Rumilly. kevin. gurcel @ orange. fr; https: // orcid. org / 0009 - 0006 - 7380 - 9072 Corresponding author. E-mail: spuissant @ ville-dijon. fr; https: // orcid. org / 0000 - 0003 - 4179 - 3636 spuissant@ville-dijon.fr Author Gurcel, Kevin 0009-0006-7380-9072 kevin.gurcel@orange.fr text Zootaxa 2023 2023-08-17 5330 4 535 560 http://dx.doi.org/10.11646/zootaxa.5330.4.4 journal article 54879 10.11646/zootaxa.5330.4.4 c089657b-b4cb-469a-9e1f-94a669a51fb1 1175-5326 8255293 8EC3E31F-ED7B-48BD-901B-1EE8AE0BAF02 Tibicina quadrisignata ( Hagen, 1855 ) pilleti Puissant ssp. n. Type material. Holotype male: Morocco , “2135 m, Près d’un ruisseau, pente rocheuse” [label rectangular, pale yellow, with black margins, printed]; “ 02/07/2010 , Taffert, 33.66235 / -4.13106 , A. François ” [label rectangular, pale yellow, with black margins, printed] and “HOLOTYPE Ô, Tibicina quadrisignata pilleti ssp. n. ” [label rectangular, red, manuscript and printed]. Paratypes (3 Ô, 1 ♀ ): Morocco 3 Ô, “ 2135 m , Près d’un ruisseau, pente rocheuse” [Label rectangular, white, with black margins, manuscript] ; “ 27-VI-2022 , Taffert, 33.66235 / -4.13106 , S. Puissant & A. François” [Label rectangular, white, with black margins, manuscript] and “ PARATYPE Ô Tibicina quadrisignata pilleti ssp. n. , [label rectangular, red, manuscript and printed]; Morocco 1 ♀ , data as the three paratypes Ô and “ PARATYPE ♀ Tibicina quadrisignata pilleti ssp. n. ” [label rectangular, red, manuscript and printed] . Other specimens used for morphological description but not include in the type series (3 Ô, 2 ♀ , 2 exuviae): Morocco 1 Ô “ Tizi n’Talghemt , 2023 m , Steppe à armoises et plantation de pins d’Alep ” [label rectangular, pale yellow, with black margins, printed]; “ 28/06/2015 , Midelt, 32.58735 / -4.55191 , A. François & B. Defaut ” [label rectangular, pale yellow, with black margins, printed] and “ Tibicina quadrisignata ssp. pilleti , Puissant & Hertach dét. 2021” [label rectangular, white, with black margins, manuscript] ; Morocco 2 Ô “Maroc 68 MA 68:27, Tizi Tfri (S Targuist), 1600 m , 27.6.68, leg. Nadig ” [label rectangular, white, with black margins, manuscript and printed] and “ Tibicina quadrisignata ssp. pilleti , Puissant & Hertach dét. 2021” [label rectangular, white, with black margins, manuscript] ; Morocco 1 ♀ “Maroc 70 MA 70: 82, Taounate-Âin , Âïcha 500–700 m , 15.7.70 leg. Nadig ” [label rectangular, white, with black margins, manuscript and printed] and “ Tibicina quadrisignata ssp. pilleti , Puissant & Hertach dét. 2021” [label rectangular, white, with black margins, manuscript] ; Morocco 1 ♀ “ 1655 m , Cédraie” [label rectangular, pale yellow, with black margins, printed]; “ 03/07/2021 , Itzer, 32.9024 / -5.22426 , A. François & G. Liénart ” [label rectangular, pale yellow, with black margins, printed] and “ Tibicina quadrisignata ssp. pilleti , Puissant & Hertach dét. 2021” [label rectangular, white, with black margins, manuscript] ; Morocco 2 exuviae from the locus typicus “ 2135 m , Près d’un ruisseau, pente rocheuse” [ Label rectangular, white, with black margins, manuscript]; “ 27-VI-2022 , Taffert, 33.66235 / -4.13106 , S. Puissant & A. François ” [Label rectangular, white, with black margins, manuscript] and “ Tibicina quadrisignata ssp. pilleti , S. Puissant dét. 2022” [label rectangular, white, with black margins, manuscript] . The holotype , one male collected by François & Defaut and one female collected by François & Liénart are deposited in the MNHN ( Paris , France ). The four paratypes and the 2 exuviae collected by Puissant & François are deposited in the Museum d’Histoire naturelle de Dijon ( MHND , Dijon , France ). The two other males and the female collected by Nadig are deposited in the MHN ( Geneva , Switzerland ) . Derivation of name. Named in honor of our late colleague Jean-Marc Pillet † ( JMP † : 1952-2010), Swiss naturalist and herpetologist who collected cicadas from Morocco over many years. Adults morphology ( Figs. 1–5 ). Head : slightly less broad than mesonotum; black except base of postclypeus brownish and lateral margins of postclypeus yellowish. Eyes brownish but greyish-bluish when specimen alive ( Fig. 5 ). Ocellus yellowish or ochraceous, the distance between lateral ocelli almost equal than the distance between each lateral ocelli and the nearest eye. Antennae blackish, supra-antennal plates yellowish. Anteclypeus black. Rostrum with the mentum brownish and the labium black. Apex of rostrum reaching base of meso-coxa. Gena and lorum blackish with dense and long light yellowish hairs. Thorax : pronotum black except an interrupted yellowish median line, yellowish marks enclosing the posterior oblique fissures, a pair of yellowish marks between both pairs of oblique fissures, the yellowish posterior margin. Mesonotum black, scutum with a pair of anterior yellow fasciae in inverted triangular shape and a pair of posterior triangular-shaped, yellow fasciae. Cruciform elevation with posterior branches yellowish and anterior branches black, mesonotal scutellar cords yellowish; wing groove yellowish. Stridulatory file, pars stridens , present on lateral angle of mesonotum. Metanotum yellowish with black anterior median margin. Metasternum blackish with a pair of lateral yellowish spots; opercula very distant from each other, black with posterior margin yellowish. Wings : fore wing hyaline, with eight apical cells; anterior and posterior cubital veins black, median and cubital veins emerging separately from the apex of the basal cell; venation mostly blackish but slightly brownish at apical cells. Hind wing with six apical cells; venation blackish to brownish, cubitus anterior vein brownish, ambient vein slightly brownish; vannus slightly smoky and its margin with anal cell brown and orange. Legs : yellowish-brownish and black. Fore legs: posterior side of coxa yellowish and anterior side black with long yellowish hairs; trochanter blackish with margin, fissures and posterior side yellowish to brownish; femora yellowish with black longitudinal bands, black edge with three spines blackish, the basal spine sharp and oblique, isolated from the two others, the second spine strong and sub-perpendicular, the apical spine very short and triangular; tibia blackish with yellowish spot on lateral sides; tarsus blackish to brownish. Mid legs: coxa yellowish except the latero-posterior side black; trochanter blackish, its base, ventral and lateral margin with spots yellowish; femora yellowish with four black longitudinal stripes of variable length, the thinner more or less interrupted; tibia black; tarsus blackish. Hind legs: similar to mid legs but lighter in colour and femora with only three black longitudinal stripes. Abdomen : tergites black, posterior margins yellowish; auditory capsules sub-circular, black, slightly convex. Tymbals bearing a basal plate which is as long as one third of the tymbal area and showing a series of eight short ribs alternating with nine long ribs, the sclerotized ribs running anterior to the tymbal plate. Sternites black with a posterior margin brownish; sternite VIII with a large blackish spot reaching the apical part, this large blackish spot bordered of yellow. Genitalia ( Figs. 1B,C,D ): pygopher curved without dorsal expansion but slightly V-shaped; uncus curved with an apex highly angular ( Fig. 1B : ↑). Claspers absent. Ventrobasal pocket of aedeagus present. Basal plate of aedeagus undivided basally and attachment to the theca by sinuation completely chitinous with no mobility. Aedeagus elongated, deeply curved and trilobed at the distal end; the two lateral lobes small and edged, strongly elongated and rounded at their apex with numerous conjunctival claws ( Fig. 1D : ↑); median lobe carrying the gonopore, vesica curved and inflated with apex short. Measurement . Seven males and three females are available. Length of body, from tip of head to end of wings in resting position: males 38.70 ± 1.26 mm (37.00–40.50), females 40.00 ± 3.16 mm (36.40–42.30). Length of body, from tip of head to end of abdomen: males 26.51 ± 1.35 mm (25.00–39.00), females 28.13 mm ± 2.04 mm (25.90–29.90). Length of head: male 2.67 ± 0.24 mm (2.40–2.90), females 2.44 ± 0.17 mm (2.30–2.63). Length of pronotum: males 4.06 ± 0.28 mm (3.70–4.50), females 3.99 ± 0.62 mm (3.30–4.48). Length of mesonotum: males 6.95 ± 0.34 mm (6.50–7.30), females 6.98 ± 0.94 mm (5.90–7.60). Length of head and thorax together: males 14.15 ± 0.77 mm (13.20–15.40), females 13.79 ± 1.64 mm (11.90–14.87). Length of abdomen: males 12.36 ± 1.18 mm (11.40–14.40), females 14.34 ± 0.84 mm (13.73–15.30). Ratio length of abdomen/head and thorax together: males 0.88 ± 0.1 mm (0.74–1.03), females 1.11± 0.08 mm (1.05–1.20). Width of head including eyes: males 8.67 ± 0.37 mm (8.00–9.12), females 8.87 ± 0.68 mm (8.10–9.40). Width of pronotum, supra-humeral plate included: males 10.63 ± 0.48 mm (10.00–11.40), females 10.90 ± 1.49 mm (9.20–12.00). Width of mesonotum: males 8.91 ± 0.49 mm (8.00–9.40), females 9.29 ± 0.81 mm (8.40–9.98). Ratio width of head including eyes/width of mesonotum: males 0.98 ± 0.02 mm (0.95–1.00), females 0.95 ± 0.04 mm (0.91–0.99). Length of fore wing: males 31.17 ± 1.40 mm (29.60–33.00), females 33.30 ± 2.62 mm (30.30–35.10). Width of fore wing: males 11.54 ± 0.66 mm (10.50–12.17), females 12.41 ± 0.79 mm (11.80–13.30). Wingspan: males 68.93 ± 1.40 mm (67.00–71.00), females 75.10 ± 7.64 mm (66.30–80.00). Ratio length of fore wing/width of fore wing: males 2.70 ± 0.09 mm (2.55–2.82), females 2.69 ± 0.18 mm (2.57–2.90). Ratio length of fore wing/width of pronotum, supra-humeral plate included: males 2.93 ± 0.11 mm (2.72–3.08), females 3.07 ± 0.21 mm (2.87–3.29). Distance between ocellus and compound eye of same side: males 1.27 ± 0.09 mm (1.10–1.35), females 1.41 ± 0.11 mm (1.30–1.52). Distance between posterior lateral ocelli: males 1.35 ± 0.09 mm (1.20–1.44), females 1.46 ± 0.21 mm (1.30–1.70). Ratio distance between ocellus and compound eye of same side/distance between posterior lateral ocelli: males 0.94 ± 0.06 mm (0.84–1.04), females 0.97 ± 0.07 mm (0.89–1.02). Exuviae ( Figs. 4A,B,C ). Very close of T . q. quadrisignata exuviae, with sturdy shape. Head with bulging postclypeus ( Fig. 4C : ↑), protruding as in some exuviae of T . q. quadrisignata . Forelimb robust with femoral comb, intermediate tooth, posterior tooth well developed. Its accessory tooth of femur ( Fig. 4B : ↑) long and flattened as in some exuviae of T . q. quadrisignata . Apical tooth of tibiae long and acuminate. FIGURE 8. Calling song recording of T. q. pilleti Puissant ssp. n. with a digitization at 96 kHz sampling frequency. Frequency amplitude envelope peaking at 28 kHz, encapsulating the second upper harmonic at around 27 kHz. Sound behavior ( Table 1 ; Figs. 8–9A,B,C,D,E,F – 10–11A,B,C,D,E ). Three males were recorded on the locus typicus by SP, then captured by SP with Alexandre François. Another male was recorded by SP without being caught. The males produce their calling song mainly from the low bushes to the tops of the trees. Densities of calling males can be very high; males usually call 1 to 3 m apart. When males emit their calling song, they raised their abdomen and lift up their wings from their body without decoupling them from the mesonotum ( Fig. 5 ). Males produce relatively short, sustained calling songs of about one minute: CD = 67.4 ± 23.5 (46.3–98.0; 4), see Table 1 . Sometimes, a small number of amplitude modulated variations is emitted at the beginning of the call ( Fig. 9C : NAV ). Group of pulses duration ( GPD ) is notably constant: 13.02 ± 0.2 ms (12.2–13.6; 346). Group of pulse period ( GPP ) is 14.1 ± 0.2 ms (13.6–14.5; 86) and groups of pulses are produced a rate ( NGP ) of 71.5 ± 0.9 s- 1 (70–73; 329). Pulses-groups ( Table 1 : NP per syllable) are made up of 8-11 pulses, typically 8 to 10 pulses per syllables, i.e. 4 to 5 pulses per tymbal. TABLE 1. Mean ± standard deviation (minimum–maximum; sample size) for the temporal and frequency parameters of T. quadrisignata pilleti and T. maldesi calling signals, with: CD, call duration of a sequence (s); GPD, group of pulses duration (ms); GPP, group of pulses period (ms); NGP, number of group of pulses per second; NP, number of pulses per group. MF, frequencies with maximum amplitude; F1, peak of the first frequency band; F2, peak of the second frequency; F3, peak of the third frequency band; *carrier frequency band. See “Materials and methods” for more explanations.

Temporal parameters		Frequency parameters
CD	GPD	GPP	NGP	NP	MF	F1	F2	F3
T. quadrisignata pilleti
Male 1	52.1	12.8 ± 0.3 (12.2–13.4; 86)	14.1 ± 0.2 (13.6–14.5; 86)	70.8 ± 0.4 (70–72; 65)	8–9	8908 ± 558 (7946–9798; 89)	*8419	9195	9690
Male 2	46.3	13.2 ± 0.2 (13.0–13.6; 85)	13.9 ± 0.2 (13.3–14.2; 91)	72.4 ± 0.5 (71–73; 80)	10–11	7867 ± 177 (7644–8871; 81)	*7817	8764	9388
Male 3	73.4	13.0 ± 0.2 (12.6–13.3; 102)	14.0 ± 0.1 (13.7–14.5; 111)	71.8 ± 0.4 (71–72; 107)	8–11	8929 ± 444 (7773–9647; 84)	8204	*9022	9561
Male 4	98	13.0 ± 0.2 (12.7–13.4; 73)	14.1 ± 0.3 (12,9–14.6; 77)	70.5 ± 0.5 (70–71; 77)	8–11	8460 ± 358 (7429–9087; 83)	8333	*8807	9561
Summary (N = 4)	67.4 ± 23.5	13.02 ± 0.2 (12.2–13.6; 346)	14.0 ± 0.2 (12.9–14.6; 365)	71.5 ± 0.9 (70–73; 329)	8–11	8553 ± 594 (7429–9798; 337)	8193 ± 266	*8947 ± 200	9550 ± 124
T. maldesi
Male 1	839.7	8.3 ± 0.4 (7.6–9.0; 100)	10.9 ± 0.6 (9.6–12.3; 121)	102.9 ± 1.4 (100–106; 103)	8–10	8593 ± 470 (7644–9367; 45)	7881	*8699	10357
Male 2	874.5	8.3 ± 0.4 (7.6–8.8; 103)	10.2 ± 0.6 (9.0–11.9; 131)	106.1 ± 0.5 (104–108; 106)	8–10	9587 ± 469 (8506–10293; 50)	8613	*9087	10034
Male 3	719.9	8.3 ± 0.3 (7.6–8.8; 98)	10.1 ± 0.5 (9.2–11.9; 125)	104.8 ± 1.4 (102–108; 107)	8–10	9172 ± 318 (8484–9668; 45)	8656	*9388	10034
Summary (N = 3)	811.4 ± 81.1	8.3 ± 0.4 (7.6–9.0; 301)	10.4 ± 0.7 (9.0–12.3; 377)	104.6 ± 1.7 (100–108; 316)	8–10	9134 ± 590 (7644–10293; 140)	8384 ± 436	*9058 ± 345	10142 ± 186

FIGURE 9. Calling song recording of T. q. pilleti Puissant ssp. n. with a digitization at 44.1 kHz sampling frequency.A, spectra (frequency vs . absolute amplitude). B, spectrogram and C, oscillogram (from top to bottom) of the beginning of the sequence. D, oscillogram of a part of C showing groups of pulses (slow amplitude modulation). E, detailed spectrogram and F, oscillogram of a part of D showing a train of groups of pulses in detail. F1, F2*, F3: three frequency peaks with F2* as carrier frequency; FFT: fast Fourier transform; GP: group of pulses (= syllable); MF: frequencies of maximum amplitude; N: number of males (N = 4) analyzed; n: number of measurements for sampling (n = 337); NAV: number of amplitude variations; light grey spectrum (at bottom): minimum amplitude spectrum plotted from the minimum amplitude value for each frequency; black spectrum (at middle): average linear amplitude spectrum plotted from the average amplitude value recorded for each frequency; grey spectrum (at top): maximum amplitude spectrum plotted from the maximum amplitude value recorded for each frequency. FIGURE 10. Spectrogram and oscillogram (from top to bottom) of the rivalry song of T. q. pilleti Puissant ssp. n. One male producing an irregular train of echemes in the foreground (M2) is interrupted by another male producing a fast trains of rivalry song pulse groups in the background (M1). Calling songs recording with digitization at 96 kHz sampling frequency (Hamming window width = 1024 points, frequency resolution = 94 Hz) shows a frequency amplitude envelope peaking at 28 kHz, encapsulating the second upper harmonic at around 27 kHz ( Fig. 8 ). To be more accurate in the frequency domain, the calling songs recording were subsequently digitized at 44.1 kHz (Hamming window width = 1024 points, frequency resolution = 43 Hz). Power spectra ( Table 1 ; Fig. 9A ) are characterized by frequencies of maximum amplitude ( MF ) having a mean of 8553 ± 594 Hz (7429–9798; 337) with three frequency peaks: a peak of the carrier frequency band (F2*) at 8947 ± 200 Hz (8764–9195; 4) with two lateral frequency bands, F1 at about 8193 ± 266 Hz (7817–9419; 4) and F3 at about 9550 ± 124 Hz (9388–9690; 4). A rivalry sound ( Fig. 10 ) has been repeatedly noted when males are perched on the same bush or tree, close to each other. The rivalry signals were produced in continuous sequences of irregular duration, corresponding more or less to the duration of the encounter. Typically, rivalry signals consist of a relatively regular succession of short trains of pulse groups. The dominant frequency F2* is around 8750 Hz for two measurements taken manually in the signal. FIGURE 11. Courtship song of T. q. pilleti Puissant ssp. n. A, spectrogram and B, oscillogram (from top to bottom) of a succession of phrases composed of two echemes. C, oscillogram of a part of B showing two phrases and their component echeme, a one long echeme (E1) and a one short terminal echeme (E2). D, detailed spectrogram and E, oscillogram of a part of C showing a phrase with, for each echeme, a low amplitude part duration (LPD) and a shorter high amplitude part duration (HPD). A courtship behavior was also recorded in one male ( Figs. 11A,B,C,D,E ). It occurred when a female arrived in the proximity of the calling male. Male courtship songs consisted of a succession of phrases composed of two echemes ( Fig. 11C : E1 and E2): a first of about 3 s duration (E1; n = 6) and a second shorter of almost 1 s duration (E2; n = 6). Each echeme ( Fig. 11E ) consists of a succession of groups of pulses arranged in two sub-groups: a low amplitude part duration ( LPD ) and a shorter high amplitude part duration ( HPD ). Echemes E1 and E2 are regrouped together and compose a phrase of about 4.3 ± 0.3 s (3.9–4.7; n = 6). During courtship behavior, fast wing-flicks preceding courtship song sequence are emitted. They produce audible clicks but were not recorded in this work. Distribution and habitat ( Figs. 14A,B : blue dot). T. q. pilleti is currently known only from Morocco , from five localities. Its geographical domain is located at the south of the nominal subspecies T. q. quadrisagnata , on the other side of the Strait of Gibraltar , Mediterranean Sea. Whereas T. q. pilleti has a restricted area distribution, T. q. quadrisignata is widely distributed throughout Western Europe and is known from France , Italy , Portugal , Spain and Switzerland . In Morocco , T. q. pilleti is present in the Mediterranean and sub-Mediterranean domains: SA3 stage, xeric semiarid class, temperate sub-class at 2025 m in sagebrush steppe; SH2 stage, xeric subhumid class, warm sub-class, at 500–700 m above sea level; SX3 and SX4 stage, sub-axeric class, temperate and cool sub-class, at 1600 m , 1655 m and 2135 m above sea level. The natural environment of the locus typicus is characterized by low but woody vegetation with a few isolated trees ( Fig. 7 ). Physical structures of the vegetation inhabit by T. q. pilleti in its area are scrub, sagebrush steppe and open wood of Aleppo pines and cedars with at least 40% of woody plants: habitat classes 4 to 8. For ecology, vegetation classes and phytoclimatic stages of T. q. quadrisignata , see Puissant (2006) and Hertach & Gerber (2020) .