Chilecicada, Sanborn, 2014

Sanborn, Allen F., Cole, Jeffrey A., Stukel, Mark, Łukasik, Piotr, Veloso, Claudio, Gonzalez, Valorie A., Karkar, Jessica B. & Simon, Chris, 2021, Thirteen new species of Chilecicada Sanborn, 2014 (Hemiptera: Auchenorrhyncha: Cicadidae: Tibicininae) expand the highly endemic cicada fauna of Chile, Zootaxa 5078 (1), pp. 1-70 : 61-64

Morphological key to the species of Chilecicada

1 Fore wing basal cell length greater than 3.95 times width..................................................... 2

- Fore wing basal cell length less than 3.95 times width........................................................ 4

2 Fore wing basal cell length about 4.00 times width, piceous fascia separating fore wing costa and radius & subcostal veins, abdominal epipleurites with central spot.................................................. C. mapuchensis n. sp.

- Fore wing basal cell length greater than 4.10 times width, no piceous fascia separating fore wing costa and radius & subcostal veins, abdominal epipleurites without central spot........................................................... 3

3 Fore wing basal cell length about 4.50 times width, fore wing length greater than 19.8 mm, body length greater than 17.5 mm, proximal basal cell lacking piceous mark, meracanthus triangular................................... C. oraria n. sp.

- Fore wing basal cell length about 4.20 times width, fore wing length less than 19 mm, body length less than 17.5 mm, proximal basal cell with piceous mark, meracanthus elongated with parallel sided base and triangular terminus.................................................................................................. C. pehuenchesensis n. sp.

4 Postclypeus with piceous longitudinal fasciae in central sulcus and along lateral margins............... C. trifascia n. sp.

- Postclypeus with piceous longitudinal fascia only in central sulcus.............................................. 5

5 Proximal basal cell without piceous mark.................................................................. 6

- Proximal basal cell with piceous mark.................................................................... 8

6 Fore wing basal cell length about 3.50 times width, fore wing length greater than 20.5 mm, piceous mark along medial opercular margin........................................................................... C. curacaviensis n. sp.

- Fore wing basal cell length less than 3.20 times width, fore wing length less than 20.5 mm, medial opercular margin lacking piceous mark........................................................................................ 7

7 Fore wing basal cell length about 2.78 times width, piceous fascia separating fore wing costa and radius & subcostal veins, meracanthus elongated with parallel sided base and triangular terminus, posteromedial opercular margin forming an obtuse angle........................................................................................................................................................................................ C. occidentis

- Fore wing basal cell length about 3.14 times width, no piceous fascia separating fore wing costa and radius & subcostal veins, meracanthus triangular, posteromedial opercular margin straight............................... C. viridicitata n. sp.

8 Body length less than 17 mm ........................................................................... 9

- Body length greater than 17 mm ........................................................................ 10

9 Fore wing basal cell length about 3.67 times width, posteromedial opercular margin straight, abdominal epipleurites with central spot...................................................................... C. citatatemporaria n. sp.

- Fore wing basal cell length about 3.00 times width, posteromedial opercular margin forming an obtuse angle, abdominal epipleurites lacking central spot.................................................... C. impartemporaria n. sp.

10 Abdominal epipleurites without central spot............................................................... 11

- Abdominal epipleurites with central spot................................................................. 12

11 Fore wing basal cell length about 3.50 times width, medial opercular margin barely medial to anteromedial margin........................................................................................ C. partemporaria n. sp.,

- Fore wing basal cell length about 3.88 times width, medial opercular margin extended to meracanthus.................................................................................................... C. trifasciunca n. sp.,

12 Fore wing basal cell length about 3.75 times width, medial opercular margin ground color, meracanthus elongated with parallel sided base and triangular terminus........................................................... C. magna n. sp.

- Fore wing basal cell length less than 3.75 times width, medial opercular margin piceous, meracanthus triangular........ 13

13 Fore wing basal cell length about 3.70 times width, fore wing length 20.75 mm or greater, terminus of male uncus bent, posteromedial opercular margin forming an obtuse angle.................................. C. parrajaraorum n. sp.

- Fore wing basal cell length about 3.50 times width, fore wing length 20.75 mm or shorter, terminus of male uncus straight, posteromedial opercular margin straight.................................................. C. culenesensis n. sp.

Bioacoustics. The fine structure of timbalization is remarkably conserved: pulse rates (MANOVA, P =7.44×10-1) and syllable rates (MANOVA, P =6.61×10-2) were not significantly different among species (character definitions illustrated in Plate 2A ). Rather, higher order temporal characters varied among Chilecicada species. Multivariate analysis of echeme duration (MANOVA, P =5.89×10 -8) and echeme rate (MANOVA, P =2.2×10 -16) produced clusters of four song types ( Fig. 15 ): type 1) medium echeme duration (0.25– 1.0 s) and slow echeme rate (0.5– 1.7 s- 1) in C. culenesensis , C. curacaviensis , C. magna , C. oraria , C. partemporaria , C. pehuenchesensis , and C. trifasciunca ; type 2) long echeme duration (>1 s) and slow echeme rate (<0.7 s- 1) in C. impartemporaria ; type 3) short echeme duration (0.07– 0.08 s) and medium echeme rate (8–10 s-1) in C. viridicitata ; and type 4) short echeme duration (<0.05 s) and fast echeme rate (>16 s-1) in C. citatatemporaria and C. occidentis . Although mean dominant frequency distributions of most species were variable, some species were divided among high ( C. impartemporaria , C. citatatemporaria , and C. viridicitata ) and low ( C. oraria and C. partemporaria ) frequency distributions (MANOVA, P =7.78×10 -8; Fig. 16 ).

Pulse rate (MANOVA, P =7.15×10 -1), syllable rate (MANOVA, P =7.39×10 -1), and mean dominant frequency (MANOVA, P =3.09×10 -1) showed no relationship with temperature. Significant temperature by species interactions were found initially for echeme duration (MANOVA, P =6.17×10 -5). When partitioned by species the C. impartemporaria echeme duration showed a positive relationship with temperature (GLM, P =4.28×10 -2, R 2 = 32.30%; Fig. 17 ). Removal of C. impartemporaria rendered the temperature by species interaction nonsignificant for the remainder of Chilecicada (MANOVA, P =3.21×10 -1).

Male Chilecicada relied on crypsis and remained stationary for long periods, rarely flying even when disturbed; C. citatatemporaria and C. impartemporaria did not attempt to fly even when captured, and C. occidentis occasionally flew short distances when disturbed but males also dropped to the ground. Chilecicada males sang in aggregations from mid morning (1043 h) to late afternoon (1758 h) and called repeatedly from the same perches. Shortly after an echeme was initiated a male would often lift the abdomen, which was correlated with an increase in echeme amplitude ( Plate 2C ). Perch height was different among sympatric C. occidentis and C. partemporaria at Peñaolén: the former perched on low bushes and forbs, while the latter tended to perch 2m or above on acacia and other trees. In addition to timbalization, wing flicks ( Plate 4 ) were observed in six Chilecicada species : C. citatatemporaria (n=1), C. impartemporaria (n=3), C. magna (n=1), C. partemporaria (n=3), C. pehuenchesensis (n=1), and C. trifasciunca (n=1). Males inserted wing flicks within the interecheme intervals of their songs. Females were observed neither within the vicinity of wing flicking males nor to flick their own wings.

Molecular phylogenetics. Molecular voucher accessions and collecting data are found in Supp. Table 1. Four independent runs resulted in identical topologies ( Fig. 18 ), and MCMC chains converged given the average standard deviation of split frequencies of 0.005146, which falls below the minimum threshold of 0.02. A monophyletic Chilecicada received 1.0 posterior probability. C. viridicitata is monophyletic and sister to all other Chilecicada . Two exemplars that fall into other clades make C. impartemporaria polyphyletic. C. curacaviensis form a grade leading up to a clade of mixed taxa.All but one C. occidentis exemplar form a clade. Two clades of C. partemporaria correspond to a geographic break: the totopotype from Santiago belongs to a clade of Santiago and Cordillera Province exemplars, while the other contains exemplars from the south in Cachapoal and Colchagüa provinces ( Fig. 19 ). Sister to the northern C. partemporaria clade is topotype C. culenesensis from Melipilla Province west of Santiago ( Fig. 19 ). Each clade consists of a single song type ( Fig. 18 ), barring the intrusion of the two wandering C. impartemporaria that bring type 2 songs into clades consisting of type 1 and type 4 songs, respectively. Genetic distance between C. viridicitata and the remainder of Chilecidada is on the order of 9.5% - 11%, for an estimated divergence time of 8-14 Ma (Supp. Table 2).

Figures

PLATE 2. Waveforms of Chilecicada songs. Left column 5 s oscillogram windows show echemes and echeme sequences, right column 0.5 s windows show pulses and syllables within echemes.

PLATE 4. Male Chilecicada wing flick behavior. Wing flicks are depicted on spectrograms with Hanning window size 512 except 4B C. impartemporaria, which best resolved on an oscillogram. Each wing flick is annotated with a dot above its position on the window, while echemes are annotated with dashes.

FIGURE 15. Chilecicada song types numbered on a scatterplot of echeme duration vs. echeme rate: type 1) medium duration, slow rate; type 2) long duration, slow rate; type 3) short duration, medium rate; type 4) short duration, fast rate.

FIGURE 16. Chilecicada songs resolved on a scatterplot of mean dominant frequency vs. echeme rate.

FIGURE 17. Positive relationship between echeme duration and ambient temperature in Chilecicada impartemporaria.

FIGURE 18. COI gene tree. Numbers at nodes are posterior probabilities. Shown to the right are 5 s calling song oscillograms. Green stars are morphologically determined vouchers, dark blue stars are molecular vouchers that were recorded, light blue stars are recordings applied from the same population as the molecular voucher, orange star is a recording applied to the terminal from another population, and red stars are possible instances of mitochondrial capture or incomplete lineage sorting.

FIGURE 19. Mapped COI gene tree. COI barcode tree from Fig. 18 visualized against a map of specimen localities using web application TreeToM (See methods). Colors represent described species.