Specificity of cicada calling songs in the genus …ljprirodm3/SueurAubinSE2003.pdf · Specificity...

Specificity of cicada calling songs in the genus Tibicina(Hemiptera: Cicadidae)

J ER OME SUEUR1 and TH IERRY AUB IN 2

1Ecole Pratique des Hautes Etudes, Biologie et Evolution des Insectes, Museum National d’Histoire Naturelle, Paris, France

and 2Neurobiologie de l’Apprentissage, de laMemoire et de la Communication – Centre National de la Recherche Scientifique,

UMR 8620, Universite Paris Sud, Orsay, France

Abstract. Males of Tibicina cicada species produce a sustained and monotonouscalling song by tymbal activity. This acoustic signal constitutes the first step inpair formation, attracting females at long range, and is involved in male–maleinteractions. The specificity of this signal was investigated for the first time forseven species and one subspecies of Tibicina occurring in France. This analysis wasachieved by describing tymbal anatomy, tymbal mechanism and calling songstructure. Male calling songs are emitted following the same general scheme:tymbals are activated alternately and the successive buckling of the sclerotizedribs that they bear produces a regular succession of groups of pulses. The struc-tural and mechanical properties shared by Tibicina species and subspecies lead to aconsiderable uniformity of the signal shape. Nevertheless, a principal componentanalysis applied to eight temporal and three frequency parameters revealed differ-ences between the signals of the species studied. In particular, calling songsdiffered in groups of pulse rate and/or in peak of the second frequency band(carrier frequency). These acoustic differences are probably linked to differences inthe numbers of tymbal ribs and body size. Groups of pulse rate and/or peak of thesecond frequency band could encode specific information. However, Tibicinacalling songs may not act as distinct specific-mate recognition systems and maynot play a leading role in the mating isolation process; rather, they might merelybelong to a complex set of specific spatial, ecological, ethological and morpho-logical characters that ensure syngamy.

Introduction

The initial step in the mating sequence of insects consists of

bringing motile males and females together in the same

place and at the same time (Alexander et al., 1997). During

this phase, both sexes usually exchange chemical, acoustical,

vibratory and/or visual signals. In many cases, one sex

produces a long-range signal encoding specific reproductive

information that is perceived and decoded by the other sex

(Bradbury & Vehrencamp, 1998). In cicadas (Hemiptera,

Cicadidae), the first stage of pair formation is achieved

essentially through sound communication, with males emit-

ting, by tymbal action, a calling song that the female

receives and uses to locate them (Alexander & Moore,

1958). Previous acoustic descriptions of these calling songs

revealed typical time and frequency patterns for each spe-

cies (Sueur, 2001). Playing a leading role in mate attraction,

long-range signals are commonly assumed to be essential in

the species-specific recognition process of cicadas. There-

fore, they can be considered to act as premating isolating

mechanisms (Dobzhansky, 1937; Mayr, 1963) or as distinct

specific-mate recognition systems (SMRSs) according to the

recognition concept of species (Paterson, 1985).

In many groups of cicada species, important acoustical

differences have been found between species that are mor-

phologically similar, such as those of Cicadetta (Popov,

1997, 1998; Puissant & Boulard, 2000) or Cicada (Simoes

et al., 2000). Acoustic similarity between closely related

species seems to be observed rarely. One such exception

could include species belonging to the Palaearctic genus

Tibicina, and some preliminary acoustic analyses have

shown that the calling songs of these species are similar

Correspondence: Jerome Sueur, Ecole Pratique des Hautes Etudes,

Biologie et Evolution des Insectes, Museum National d’Histoire

Naturelle, 45 rue Buffon, F-75005 Paris, France. Tel.:þ33 1 40 79 31 57;

fax:þ33 1 40 79 36 99; e-mail: [email protected]

Systematic Entomology (2003) 28, 481–492

# 2003 The Royal Entomological Society 481

(Fonseca, 1991, 1996; Boulard, 1995). Such similarity

suggests that calling songs of Tibicina are not involved in

species recognition. However, are the Tibicina calling songs

really similar? We have tried to answer this question by

conducting anatomical and acoustical analyses on seven

species and one subspecies of Tibicina present in the south

of France. The tymbal system of each species was examined

first through structural observations and mechanical experi-

ments. Then, the calling songs of the different taxawere analysed

in the time and frequency domains to determine if fine details of

acoustic structure could encode specific information.

Materials and methods

Subjects and location

Seven species and one subspecies of Tibicina were

recorded in France, Switzerland, Spain and Portugal

(Table 1). All the recordings were made in the field during

June and July 1999–2001. Calling songs of T. steveni

(Krynicki, 1837) recorded by Jean-Marc Pillet (Switzerland,

22 June 2000), Andrej Popov (Georgia, 1 July 1976) and

Stephane Puissant (France, 6 July 2002) were added to our

data to increase the number of signals analysed.

Recording procedure

Recordings were made using a Telinga Pro4PiP micro-

phone (Telinga Microphones, Tobo, Sweden) (frequency

response 40–18 000Hz� 1 dB) connected to a Sony

TCD-D8 digital audiotape recorder (sampling frequency

44.1 kHz, frequency response flat within the range

20–20 000Hz). The recordings were carried out between

11.00 and 18.00 hours, a period corresponding to the

maximal activity of cicadas. The ambient temperature

ranged from 25 to 35 �C with a mean of 31 �C. Additional

recordings by J.-M. Pillet, A. Popov and S. Puissant were

made with similar equipment.

Tymbal anatomy and activity

The morphology of tymbals was described for each

species. In particular, the number of short and long ribs

borne on each tymbal was analysed. To determine whether

tymbal muscles contract alternately or synchronously

during sound production, the distress song produced when

males were handled was recorded with both tymbals intact

or with one tymbal destroyed. The tymbal was destroyed by

an anterior–posterior incision made with a microscalpel.

The rate at which one tymbal muscle contracted was then

determined from oscillograms. In another experiment,

tymbal muscles of animals were exposed by removing the

wings and the abdomen at the third abdominal segment.

The intact tymbal was artificially activated by pulling on the

tymbal muscle apodeme with a thin pair of forceps. Record-

ings were made when all ribs were buckled artificially and

the first long rib alone was twitched artificially.

Signal analysis

Signals were digitized from the analog output of the DAT

recorder at a sampling rate of 32 kHz and then analysed in

both temporal and frequency domains using the SYNTANA

analytical package (Aubin, 1994).

Tibicina calling songs showed a common general design:

a succession of groups of pulses arranged in two subgroups.

Two categories of amplitude modulations were distinguish-

able: a slow amplitude modulation distributed within

groups of pulses and a fast amplitude modulation at the

level of each pulse. In the frequency domain, the power

spectrum of all the signals was characterized by three

main peaks (F1, F2, F3). These frequency peaks were a

by-product of the fast amplitude modulation detected in

the pulses. F1 and F3 correspond to the two lateral bands,

and F2 to the carrier frequency, such that 1/(F2�F1) or

1/(F2�F3) correspond to the modulation rate of pulses.

Table 1. Taxa studied and recording sites. Records of T. steveni in

Switzerland and France refer to Sueur et al. (2003).

Site Region Country

T. corsica corsica (Rambur)

Galeria Haute Corse (2A) France

Monticello Haute Corse (2A) France

Occhiatana Haute Corse (2A) France

Barcaggio Haute Corse (2A) France

T. corsica fairmairei Boulard

Site 1 Herault (34) France


T. garricola Boulard

Domazan Gard (30) France

Serignan-du-Comtat Vaucluse (84) France

Sesimbra Baixo Alentejo Portugal

T. haematodes (Scopoli)

Cairanne Vaucluse (84) France

T. nigronervosa Fieber

Santo-Pietro-di-Tenda Haute Corse (2A) France

Barcaggio Haute Corse (2A) France

T. quadrisignata (Hagen)

Molitg-les-Bains Pyrenees-Orientales (66) France

Sournia Pyrenees-Orientales (66) France

Campoussy Pyrenees-Orientales (66) France

Sainte-Maxime Var (83) France

T. steveni (Krynicki)

Martigny-Croix Valais Swiss

Martigny-Combe Valais Swiss

Fully Valais Swiss

Castelneau-de-Montmirail Tarn (81) France

Cherkodzi – Georgia

T. tomentosa (Olivier)



Site 3 Var (83) France

Mertola Distrito de Beja Portugal

Valencia de Alcantara Provencia de Caceres Spain

482 J. Sueur and T.Aubin

# 2003 The Royal Entomological Society, Systematic Entomology, 28, 481–492

In the temporal domain, call duration (CD), silence

intercall duration (ICD), group of pulses duration (GPD),

group of pulses period (GPP), number of group of pulses

per second (NGP), and number of pulses per group (NP)

were measured (Fig. 1). Cicada males can produce wing-

clicking before they emit their calling song (Boulard,

1990). They can also produce important amplitude

variations at the beginning of sound emission and short

interruptions (< 1 s) during sound production. Therefore,

presence or absence of wing-clicking (WC) at the beginning

of the calling song was noted. The number of amplitude-

modulated variations (NAV) at the beginning of the call

and the number of short gaps (NSG) interrupting the

calling songs were also counted.

In the frequency domain, the three main bands (F1, F2,

F3) were analysed on spectra computed with a fast Fourier

transformation (FFT), using a 512 point window size

(Df¼ 62.5Hz) with 50% overlap at a 32-kHz sampling

rate (Fig. 1).

Because of occasional background noise, fine temporal

analysis (GPD, GPP, NGP, NP) and frequency parameters

(F1, F2, F3) were not available for all males recorded.

Because frequency parameters might be correlated to the

size of the sound source (Bennet-Clark & Young, 1994;

Bennet-Clark, 1998), mean of body length (BL) was calcu-

lated for each taxon and then correlated with F1, F2 and F3.

Statistical analysis

A two-factor principal components analysis (PCA) was

computed first using all temporal and frequency

parameters. This allowed us to identify potential correl-

ations between variables and led us to investigate which

temporal or frequency parameters better explain interspecific

variation. These parameters were analysed with an analysis

of variance (ANOVA) using taxon as the independent variable.

Post-hoc HSD Tukey’s tests for samples with unequal sizes

were used to compare taxon means for significant differences

(Spjotvoll & Stoline, 1973). The significance of the correl-

ation between BL and F1, F2 and F3 was tested using

Spearman’s rank test (Scherrer, 1984). All statistical analyses

were performed using STATISTICA v. 5.1 (StatSoft France,

1996).

Results

Tymbal anatomy and activity

The anatomy of the tymbal of Tibicina species was similar

to that already described for T. haematodes by Sueur &

Aubin (2002a) (Fig. 2). The tymbal muscle was inserted in

the dorsal part of the posterior tymbal plate. A series of

sclerotized ribs ran anterior to the tymbal plate. These long

ribs are orientated dorsoventrally, and are narrower and

more highly sclerotized at their centres. Short ribs alternate

with long ribs. The number of long ribs differed between

species (Table 2). The number of long ribs was asymmetric

(�1) between left and right tymbals in 8.62% of males

examined. A dorsal bar connects long ribs 1–4 or 1–5 of

the T. haematodes tymbal, long ribs 1–4 of the T. tomentosa

tymbal, and all long ribs in the remaining taxa.

Distress songs of Tibicina with both tymbals intact

showed a regular succession of groups of pulses. Each

group of pulses was divided into two subgroups. Distress

songs with one tymbal destroyed were made up of only one

of these subgroups (Fig. 3). This indicated that tymbal

muscles contracted alternately and that each muscle

contraction produces a subgroup of pulses. Thus, each

subgroup of pulses was correlated with the activation of

one tymbal. Manual manipulations of one tymbal showed

F2

F1

0

16

8

0

A

B

C

D

Abs

olut

e am

plitu

de (

linea

r sc

ale)

4000 8000Frequency (Hz)

Freq

uenc

y (k

Hz)

Abs

olut

e am

plitu

de

12000 16000

11.4 s

groups of pulses (slow AM) (fast AM)

sub-groups of pulses

single pulse (fast AM)

0.4 s

0.1 s

0.03 s

F3

Fig. 1. Calling song of T. garricola. A, Spectrum (frequency vs.

absolute amplitude), spectrogram and oscillogram (from top to

bottom) of the beginning of the sequence; B, detailed oscillogram

showing groups of pulses; C, detailed oscillogram showing

subgroups of pulses; D, detailed oscillogram showing pulses.

Specificity of cicada calling songs 483


the production of successive impulses during inward move-

ment (IN) and a single faint pulse during outward move-

ment (OUT) (Fig. 4). We can thus infer that the sound

production of Tibicina species studied is produced following

the scheme IN1 – (OUT1)&IN2 – (OUT2), where subscripts

1 and 2 represent the separate tymbals. The activation of

one rib produced one in-pulse and one out-pulse. The suc-

cession of pulses was thus correlated with the successive

buckling of long ribs.

Signal analysis

Measurements of temporal parameters are summarized

in Table 3 and compared in Figs 5–7. Some general fea-

tures can be recognized first: T. haematodes differed from

other taxa by short CD values and T. tomentosa differed

from other taxa by high NGP values and low GPD, GPP

and NP values. The two subspecies T. c. corsica and

T. c. fairmairei had a similar temporal pattern, except

that NP appeared to be slightly higher in T. c. fairmairei

than in T. c. corsica, although the difference is not significant

statistically (HSD Tukey’s test for samples with unequal sizes,

P>0.1). In addition,T. haematodesproduced thehighestNAV

whereasT. tomentosawas the species that interrupted its calling

song sequence by the highest NSG. Finally, T.garricola

and T. steveni (S. Puissant, personal communication) were the

only two taxa that produced wing-clicking before calling

song production.

Measurements of frequency parameters are summarized

in Table 4 and compared in Fig. 8. Three groups of taxa can

be identified easily: (1) taxa producing ‘low’ frequencies, i.e.

T. haematodes and T. steveni; (2) taxa producing ‘middle’

frequencies, i.e.T. garricola,T. quadrisignata andT. tomentosa;

and (3) taxa producing ‘high’ frequencies, i.e. T. c. corsica,

T. c. fairmairei and T. nigronervosa.

Body lengths and frequency peaks of F1, F2 and F3 were

negatively correlated as shown in Fig. 9 (Spearman correl-

ation rank test, P< 0.01). T. tomentosa appeared as an

exception, because it produced relatively low frequencies

compared with its small body length.

Statistical analysis

Principal components analysis (PCA) of calling song vari-

ation among species is shown in Table 5 and Figs 10 and 11.

Factor 1 explained 43.3% of the total variance and factor 2

explained 21.3%. Species and subspecies were well sep-

arated, as were distant populations of T. garricola. Table 5

and Fig. 10 show that measurements CD, ICD, NAV, NSG

and WC did not contribute significantly to either factor. In

addition, F1, F2 and F3 appeared to be highly correlated, as

were GPD and GPP. Therefore, the significant measure-

ments that can be retained to differentiate among Tibicina

calling songs are F2 (or F1 or F3), GP (or GPP), NGP and

NP. In fact, as shown in Fig. 12, Tibicina calling songs of

the different species can be differentiated using only NGP

and F2, in combination.

NGP and F2 differed between taxa (ANOVA, F18,162¼367.3, P¼ 0). The results of comparisons of NGP and F2

between the different taxa (post hoc HSD Tukey’s test for

samples with unequal sizes) are presented in Table 6.

Except T. c. corsica and T. c. fairmairei, taxa showed

pairwise differences for at least one measurement. The

lowest differences for taxa with overlapping geographical

distributions (Sueur & Puissant, 2002) were observed

between: (1) T. nigronervosa and T. c. corsica that

differed for NGP (P< 0.001) but not for F2 (P> 0.5);

Fig. 2. Representation of a tymbal of T. garricola. Abbreviations:

MI, muscle insertion; LR, long rib; SR, short rib; TP, tymbal plate.

Table 2. Mode, minimum and maximum values, and sample size

(n) for the number of tymbal long ribs for each taxon of Tibicina

studied.

Taxon Mode Min.–max. n

T. corsica corsica 10 10–11 17

T. corsica fairmairei 11 10–11 16

T. garricola 10 9–10 17

T. haematodes 7 7–8 19

T. nigronervosa 8 8–9 18

T. quadrisignata 9 8–9 17

T. steveni 9 8–10 8

T. tomentosa 8 7–8 21



(2) T. garricola and T. c. fairmairei that differed for F2

(P< 0.001) but not for NGP (P> 0.5); (3) T. garricola

and T. quadrisignata that differed for NGP (P< 0.001) but

not for F2 (P> 0.5); (4) T. steveni and T. haematodes that

differed for NGP (P< 0.001) but not for F2 (P> 0.5).

Furthermore, distant populations of T. garricola differed

for NGP (P< 0.001) but not for F2 (P> 0.5). As already

reported by Sueur & Puissant (2003), distant populations

of T. tomentosa differed for NGP (P< 0.001) but not for

F2 (P> 0.1).

Discussion

Tibicina cicada species are probably the most difficult Medi-

terranean species to identify acoustically in the field. In

most cases, their sustained and monotonous calling songs

do not allow a human listener to distinguish among them.

Nevertheless, our acoustic description identifies some fine

differences between the seven species and the one subspecies

found in France and some differences were detected even at

population level.

Calling songs of Tibicina species share the same general

pattern, consisting of a regular succession of groups of

pulses with three main frequency bands. Assuming Tibicina

to be a monophyletic group, this common pattern could be

considered as a synapomorphic ethological character for

the genus and then could constitute a good candidate for

generic identification. By contrast, calling songs of Tibicina

species mainly differ in the number of groups of pulses

produced per second and in the frequency values of the

pulses. Intraspecific differences have also been found

between distant populations of both T. garricola and

T. tomentosa. The differences between T. garricola populations

could not be interpreted as a subspecific differentiation because

intermediate values could exist for populations distributed

betweenFrance andwesternPortugal. Similarly, thedifferences

observed between T. tomentosa populations were not signifi-

cant enough to divide the population into two distinct taxa, as

already discussed (Sueur & Puissant, 2003).

Which factors could then explain simultaneously the

general similarities and the particular differences observed

between species, subspecies or populations? All taxa studied

here show the same general morphology: the lateral tymbal

covers were lacking, the ventral opercula, short in length,

have a similar shape, and the abdomens also show the same

configuration, being half filled with the air sacs (Sueur,

2002). In particular, no obvious anatomical differences

were found in the tymbal structures: they all exhibit the

same alternation of long and short sclerotized ribs. In addi-

tion, our experiments on tymbal activity show that all the

Fig. 3. Tymbal activity: distress song

of T. garricola emitted with (A) both

tymbals intact and (B) with one tymbal

destroyed.

16

Freq

uenc

y (k

Hz)

Abs

olut

e am

plitu

ide

Abs

olut

e am

plitu

ide

8

0

16

Freq

uenc

y (k

Hz)

8

0

0

0

IN

IN

OUT

0.30 s

OUT

0.08 s

r1

r1

r2 r3r4r5r6

Fig. 4. Tymbal activity: manual activation of the tymbal of

T.garricola, with (A) all ribs buckled and (B) first rib alone buckled.



males produce their calling songs following the same

process as already described for species belonging to

Tibicina (Popov, 1975; for T. steveni and not T. intermedia

Fieber; Fonseca, 1991, 1996, for T. garricola and not

T. quadrisignata). All these common morphological and

mechanical features certainly contribute to the similarity

observed for Tibicina calling songs. In addition, the sound

production mechanism seems to be similar to those of

Abricta curvicosta (Germar) (Young, 1972; Young &

Josephson, 1983a) and Magicicada cassini (Fischer) (Reid,

1971; Young & Josephson, 1983b). Lyristes linnei (Smith &

Grossbeck) (Hennig et al., 1994), Cyclochila australasiae

(Donovan) (Young & Bennet-Clark, 1995; Bennet-Clark,

1997) and Cystosoma saundersii Westwood (Simmons &

Young, 1978; Young, 1980; Bennet-Clark & Young, 1998)

also show the same basic tymbal activation and structure,

but sounds produced by successive ribs are fused in one or

two pulses.

By contrast, we found differences between Tibicina

species in the number of tymbal ribs and also in male

body lengths, both of which are correlated with the signal

structure. First, differences in the number of ribs cause

differences in the number of pulses per groups of pulses

and then modify the duration of these groups of pulses.

Fig. 5. Mean (�xx) and SD for group of pulse duration (GPD) and

group of pulses period (GPP).

Fig. 6. Mean (�xx) and SD for number of pulses per group of pulses (NP). Table3.Average(� xx),SD

andsamplesize

forthetemporalparametersofTibicinacallingsignals.Abbreviations:CD,callduration;IC

D,silence

intercallduration;GPD,groupofpulses

duration;GPP,groupofpulses

period;NGP,number

ofgroupofpulses

per

second;NP,number

ofpulses

per

group;WC,presence

orabsence

ofwing-clicking;NAV,number

of

amplitudevariations;NSG,number

ofshort

gaps;F,France;P,Portugal;S,Spain

Species

CD

ICD

GPD

GPP

NGP

NP

NAV

NSG

WC

T.c.corsica

52.8�35.1

(3)

112.9�96.8

(3)

13.8�0.5

(4)

15.8�0.5

(4)

62�

2(4)

11.8�0.5

(4)

00.28�0.4

(4)

0

T.c.fairmairei

81.5�82.3

(12)

63.2�53.1

(12)

13.7�0.6

(13)

15.9�0.3

(13)

62�1.3

(13)

13.2�0.7

(13)

00.26�0.58(14)

0

T.garricola

(F)

42.2�34.7

(11)

76.4�202.6

(11)

13.3�0.5

(13)

14.9�0.6

(13)

67.2�1.3

(13)

10.9�0.8

(13)

00.09�0.30(15)

1

T.garricola

(P)

61.2�73.2

(3)

399.3�509.9

(3)

16.6�0.9

(5)

18.8�0.5

(4)

55.6�1.5

(5)

12.0�

0(5)

00

1

T.garricola

(FþP)

48.8�25.4

(14)

126.7�156.6

(14)

14.2�1.6

(18)

15.9�1.9

(18)

64�5.5

(18)

11.2�0.9

(18)

00.07�0.26(20)

1

T.haem

atodes

14.0�2.3

(19)

12.8�8.7

(19)

8.2�0.4

(11)

10.2�0.4

(11)

98.3�1.6

(11)

8.0�0.6

(11)

3.66�1.38(14)

0.19�0.32(21)

0

T.nigronervosa

72.1�60.1

(7)

28.4�13.1

(7)

7.9�0.4

(8)

8.9�0.4

(8)

108.6�1.1

(8)

9.0�0.5

(8)

0.33�0.71(8)

0.06�0.18(8)

0

T.quadrisignata

93.8�66.8

(10)

280.8�241.9

(8)

12.1�0.7

(16)

13.9�0.6

(16)

74.1�2.16(16)

10.0�1.0

(16)

0.46�0.83(17)

0.24�0.27(17)

0

T.steveni

186�187(7)

–12.3�1.3

(4)

16.8�1.0

(4)

58.5�3.9

(6)

9.8�0.5

(4)

00

1

T.tomentosa

(F)

50.0�28.1

(6)

163.6�87.1

(5)

4.7�6.8

(9)

6.8�0.4

(9)

152.9�6.1

(9)

6.8�0.4

(9)

01.18�1.17(12)

0

T.tomentosa

(SþP)

76.0�27.6

(4)

452.8�154.5

(4)

4.3�0.8

(7)

6.3�0.5

(7)

167.6�10.0

(7)

6.0�1.2

(7)

01.07�1.24(6)

0

T.tomentosa

(FþSþP)

60.3�29.6

(10)

292.1�199.5

(9)

4.5�0.7

(16)

6.6�0.5

(16)

159.3�10.8

(16)

6.4�0.9

(16)

01.15�1.16(18)

0



Second, small differences in the physiological and

mechanical properties of tymbal muscles, not revealed by

our investigations, could also generate fine differences in

the number of groups of pulses produced per second.

Third, differences in body length could be responsible

for the differences observed in the frequency bands of

the signals emitted by the different species. Such a rela-

tionship between body length and frequency of calling

song has been documented already for several cicada

species (Bennet-Clark & Young, 1994). Tibicina tomentosa

produces relatively low-frequency song compared with its

short body length. Such a failure in the body length/signal

frequency relationship has been reported already for

Magicicada septendecim (L.) and interpreted in terms of

the thickness of the tympana (Bennet-Clark & Young,

1992). Further morphological studies measuring the

thickness of tympana are needed to look for possible

differences between T. tomentosa and the other Tibicina

studied. Hence, fine differences in morphology and

physiology could engender fine differences in the signal

properties. Such factors have been identified already as

potential forces acting on signal structure and evolution

(Endler, 1992; Forrest, 1994).

We have therefore identified some parameters of sexual,

or fertilization, signals that show small amounts of inter-

specific variation. Such acoustic parameters, mainly the

number of groups of pulses per second and the peak of

the second frequency band, could encode species-specific

information, as behavioural components of distinct

SMRSs (Den Hollander, 1995; Lane, 1995; Villet, 1995).

In Cystosoma saundersii, identification of conspecific

males by females at long range is based only on analysis

of carrier frequency (Doolan & Young, 1989). In Cicada

barbara lusitanica, only rough temporal song structure has

been suggested to be important for song discrimination in

long-range communication (Fonseca & Revez, 2002). A

specific recognition process based simultaneously on fine

frequency and time parameters is indeed quite rare in insects

(Hennig & Weber, 1997). In addition, previous playback

studies using T. haematodes have shown that males were

Fig. 7. Mean (�xx) and SD for number of groups of pulses per

second (NGP).

Fig. 8. Mean (�xx) and SD for peak of first frequency band (F1),

second frequency band (F2) and third frequency band (F3).

Table 4. Average (�xx), SD and sample size for the frequency parameters of Tibicina calling signals. Abbreviations: F1, peak of the first

frequency band; F2, peak of the second frequency band; F3, peak of the third frequency band; F, France; P, Portugal; S, Spain.

Taxon F1 F2 F3

T. c. corsica 8645� 71 (3) 9782� 96 (3) 11081� 101 (3)

T. c. fairmairei 8819� 101 (11) 10029� 131 (11) 11177� 115 (11)

T. garricola (F) 7587� 170 (13) 8529� 124 (13) 9551� 216 (13)

T. garricola (P) 7439� 1168 (5) 8731� 219 (5) 9715� 152 (5)

T. garricola (FþP) 7545� 177 (18) 8585� 175 (18) 9596� 210 (18)

T. haematodes 6641� 218 (9) 7516� 247 (9) 8406� 170 (9)

T. nigronervosa 8620� 101 (8) 9806� 189 (8) 11330� 105 (8)

T. quadrisignata 7503� 113 (10) 8614� 188 (10) 9651� 117 (10)

T. steveni 6416� 234 (8) 7221� 148 (8) 8229� 270 (8)

T. tomentosa (F) 7091� 328 (4) 8299� 219 (4) 9407� 379 (4)

T. tomentosa (SþP) 7214� 88 (4) 8416� 81 (4) 9626� 177 (4)

T. tomentosa (F þSþP) 7153� 231 (8) 8357� 165 (8) 9156� 297 (8)



unable to decode such fine time or frequency parameters,

and they reply even to the calling songs of other species of

Tibicina (e.g. T. c. fairmairei, T. garricola, T. quadrisignata

and T. tomentosa). However, they do not reply to the calling

songs produced by species of other genera showing great

acoustic differences (e.g. Cicada orni L., Cicadatra atra

(Olivier)) (Sueur & Aubin, 2002a, b). Furthermore, fine

temporal parameters of such long-range signals probably

are altered during propagation at great distance (>8m)

through numerous natural obstacles (Wiley & Richards,

1978). Females are probably also unable to discriminate at

long range the calling songs of the different species of

Tibicina, and the specific differences observed here could

be considered as epiphenomena of morphological and ana-

tomical contingencies. In this way, Tibicina calling songs

could not be considered as distinct SMRSs, instead merely

as playing a general role in long-range attraction. When

males and females are at close range, other signals, such as

acoustic courtship signals or chemical, visual or vibrational

signals (Claridge et al., 1999), could play a role in the

Tibicina fertilization system and could act as distinct

SMRSs. Besides, as documented already for crickets

(Walker, 1974) and other cicada species (Alexander &

Moore, 1962) with similar calling songs, other parameters

may maintain species isolation. It is particularly true that

geographical and ecological factors reduce temporal and

spatial overlap between Tibicina taxa in France (Sueur &

Puissant, 2002). To conclude, syngamy in the Mediterra-

nean Tibicina species is probably the result of the combined

action of spatial, ecological, morphological and ethological

factors (among which are acoustic signals), rather than of

only one factor.

Fig. 9. Correlation between body length

(BL) and frequency peaks (F1, F2, F3). cor,

T. c. corsica; fai, T. c. fairmairei; gar, T.garri-

cola; hae,T. haematodes; nig,T.nigronervosa;

ste, T. steveni; tom, T. tomentosa. F1¼14 535.2� 286.083*BL; F2¼ 16 886.06�339.347*BL;F3¼ 19049.24� 383.93*BL.

Fig. 10. Scatterplot of the factor loadings

of a principal components analysis (PCA)

of time and frequency measurements.

Variable names are given in Tables 3 and 4.



Acknowledgements

We are grateful to Michel Boulard and Thierry Bourgoin

for continual support. We are indebted to Jean-Marc Pillet,

Andrej Popov and Stephane Puissant for kindly providing

recordings of T. steveni. We thank Pierre Teocchi, Laetitia

Teocchi, Jacques Coffin, Dominique Cerqueira, Robert

Germain, Stephane Puissant and Nadine Fille for their

help during our visits to the south of France. We are

indebted to Henry C. Bennet-Clark for critical reading of

the manuscript. We are also grateful to Charles S. Henry

and two anonymous referees for comments and improve-

ment of the English. This study was partially conducted in

the ‘Harmas de Jean-Henri Fabre’ (Museum National

d’Histoire Naturelle, France).

Table 5. Factor loadings of the first two principal components from a principal components analysis (PCA) of time and frequency parameter

measurements. Asterisks indicate loadings where the correlation coefficient R > 0.700. For abbreviations see Tables 3 and 4.

Variable Factor 1 Factor 2

Frequency parameters

F1 � 0.786* 0.561

F2 � 0.769* 0.604

F3 � 0.714* 0.663

Temporal parameters

CD � 0.182 0.223

ICD 0.044 0.010

GPD � 0.853* � 0.470

GPP � 0.833* � 0.482

NGP 0.809* 0.502

NP � 0.935* � 0.124

NSG 0.388 0.356

Other parmeters

NAV 0.508 � 0.427

WC � 0.293 � 0.595

Fig. 11. Scatterplot of the first two factors of a principal components analysis (PCA) of time and frequency measurements. Each data point

represents a single individual, coded by species. Variable names are given in Tables 3 and 4. F, France; P, Portugal; S, Spain.



References

Alexander, R.D., Marshall, D.C. & Cooley, J.R. (1997) Evolutionary

perspectives on insect mating. The Evolution of Mating Systems in

Insects and Arachnids (ed. by J. C. Choe & B. J. Crespi), pp. 4–31.

Cambridge University Press, Cambridge, U.K.

Alexander, R.D. & Moore, T.E. (1958) Studies on the acoustical

behaviour of seventeen-year cicadas (Homoptera: Cicadidae:

Magicicada). Ohio Journal of Science, 58, 107–127.Alexander, R.D. &Moore, T.E. (1962) The evolutionary relationships

of 17-year and 13-year cicadas, and three new species (Homoptera,

Cicadidae: Magicicada). Miscellaneous Publications of the Museum

of Zoology, University of Michigan, 121, 1–59.

Fig. 12. Scatterplot of the number of group of pulses per second (NGP) and the peak of the second frequency band (F2) values. Each data

point represents a single individual, coded by species. F, France; P, Portugal; S, Spain.

Table 6. Between-species comparison for the number of groups of pulses (NGP) and the peak of the second frequency band (F2), using

post hoc HSD Tukey’s test for samples with unequal sizes.

T. c. T. c. T. T. T. T. T.

Taxon corsica fairmairei garricola haematodes nigronervosa quadrisignata steveni Measurements

T. c. fairmairei ns 1 NGP

ns 1 F2

T. garricola ns ns 1 NGP

** ** 1 F2

T. haematodes ** ** ** 1 NGP

** ** ** 1 F2

T. nigronervosa ** ** ** ** 1 NGP

ns ns ** ** 1 F2

T. quadrisignata * ** ** ** ** 1 NGP

** ** ns ** ** 1 F2

T. steveni ns ns ns ** ** ** 1 NGP

** ** ** ns ** ** 1 F2

T. tomentosa ** ** ** ** ** ** ** NGP

** ** ns ** ** ns ** F2

ns, P > 0.1; *P < 0.5; **P < 0.001.



Aubin, T. (1994) Syntana: a software for the synthesis and analysis

of animal sounds. Bioacoustics, 6, 80–81.Bennet-Clark, H.C. (1997) Tymbal mechanics and the control of

song frequency in the cicada Cyclochila australasiae. Journal of

Experimental Biology, 200, 1681–1694.Bennet-Clark, H.C. (1998) Size and scale effects as constraints in

insect sound communication. Philosophical Transactions of the

Royal Society of London, 353, 407–419.Bennet-Clark, H.C. & Young, D. (1992) A model of the mechanism

of sound production in cicadas. Journal of Experimental Biology,

173, 123–153.Bennet-Clark, H.C. & Young, D. (1994) The scaling of song

frequency in cicadas. Journal of Experimental Biology, 191,

291–294.Bennet-Clark, H.C. & Young, D. (1998) Sound radiation by the

bladder cicada Cystosoma saundersii. Journal of Experimental

Biology, 201, 701–715.Boulard, M. (1990) Contributions a l’entomologie generale et

appliquee. 2. Cicadaires (Homopteres Auchenorhynques).

Premiere partie: Cicadoidea. Ecole Pratique des Hautes Etudes,

Biologie et Evolution des Insectes, 3, 55–244.Boulard, M. (1995) Postures de cymbalisation, cymbalisations et

cartes d’identite acoustique des Cigales. 1. Generalites et especes

mediterraneennes. Ecole Pratique des Hautes Etudes, Biologie et

Evolution des Insectes, 7/8, 1–72.Bradbury, J.W. & Vehrencamp, S.L. (1998) Principles of Animal

Communication. Sinauer Associates, Sunderland, MA.Claridge, M.F., Morgan, J.C. & Moulds, M.S. (1999) Substrate-

transmitted acoustic signals of the primitive cicada Tettigarcta

crinita Distant (Hemiptera Cicadoidea, Tettigarctidae). Journal

of Natural History, 33, 1831–1834.Den Hollander, J. (1995) Acoustic signals as specific-mate

recognition signals in leafhoppers (Cicadellidae) and planthop-

pers (Delphacidae) (Homoptera: Auchenorrhyncha). Speciation

and the Recognition Concept. Theory and Application (ed. by

D. M. Lambert & H. G. Spencer), pp. 440–463. Johns Hopkins

University Press, Baltimore, MD.Dobzhansky, T. (1937) Genetics and the Origin of Species.

Columbia University Press, New York.Doolan, J.M. & Young, D. (1989) Relative importance of song

parameters during flight phonotaxis and courtship in the

bladder cicada Cystosoma saundersii. Journal of Experimental

Biology, 141, 113–131.Endler, J.A. (1992) Signals, signal conditions, and the direction of

evolution. American Naturalist, 139, s125–s153.Fonseca, P.J. (1991) Characteristics of the acoustic signals in nine

species of cicadas (Homoptera, Cicadidae). Bioacoustics, 3,

173–182.Fonseca, P.J. (1996) Sound production in cicadas: timbal muscle

activity during calling song and protest song. Bioacoustics, 7,

13–31.Fonseca, P.J. & Revez, M.A. (2002) Song discrimination by male

cicadas Cicada barbara lusitanica (Homoptera, Cicadoidea).

Journal of Experimental Biology, 205, 1285–1292.Forrest, T.G. (1994) From sender to receiver: propagation and

environmental effects on acoustic signals. American Zoologist,

34, 644–654.Hennig, R.M. & Weber, T. (1997) Filtering of temporal parameters

of the calling song by cricket females of two closely related

species: a behavioral analysis. Journal of Comparative Physiology

A, 180, 621–630.Hennig, R.M., Weber, T., Moore, T.E., Kleindienst, H.-U. &

Popov, A.V. (1994) Function of the tensor muscle in the cicada

Tibicen linnei. Journal of Experimental Biology, 187, 33–44.

Lane, D.H. (1995) The recognition concept of species applied in

an analysis of putative hybridization in New Zealand cicadas of

the genus Kikihia (Insecta: Hemiptera: Tibicinidae). Speciation



University Press, Baltimore, MD.Mayr, E. (1963) Animal Species and Evolution. Belknap Press of

Harvard University Press, Cambridge, MA.Paterson, H.E.H. (1985) The recognition concept of species.

Species and Speciation (ed. by E. S. Vrba), pp. 21–29. Transvaal

Museum Monograph 4. Transvaal Museum, Pretoria.Popov, A.V. (1975) The structure of timbals and characteristic of

sound signals of singing cicadas (Homoptera, Cicadidae) from the

southern regions of the USSR. Entomological Review, 54, 7–35.Popov, A.V. (1997) Acoustic signals of the three morphologically

similar species of singing cicadas (Homoptera: Cicadidae).

Entomological Review, 76, 1–11.Popov, A.V. (1998) Sibling species of the singing cicadas Cicadetta

prasina (Pall.) and C. pellosoma (Uhler) (Homoptera: Cicadidae).

Entomological Review, 78, 309–318.Puissant, S. & Boulard, M. (2000) Cicadetta cerdaniensis, espece

jumelle de Cicadetta montana decryptee par l’acoustique. Ecole

Pratique des Hautes Etudes, Biologie et Evolution des Insectes, 13,

111–117.Reid, K.H. (1971) Periodical cicada: mechanism of sound

production. Science, 172, 949–951.Scherrer, B. (1984) Biostatistique. Gaetan Morin, Montreal.Simmons, P.J. & Young, D. (1978) The tymbal mechanism and

song patterns of the bladder cicada, Cystosoma saundersii.

Journal of Experimental Biology, 76, 27–45.Simoes, P.C., Boulard, M., Rebelo, M.T., Drosopoulos, S.,

Claridge, M.F., Morgan, J.C. & Quartau, J.A. (2000) Differ-

ences in the male calling songs of two sibling species of Cicada

(Hemiptera: Cicadoidea) in Greece. European Journal of

Entomology, 97, 437–440.Spjotvoll, E. & Stoline, M.R. (1973) An extension of the T-method

of multiple comparison to include the cases with unequal sample

sizes. Journal of the American Statistical Association, 68,

976–978.StatSoft France (1996) STATISTICA Pour Windows [Manuel de

Programmation]. StatSoft, Charenton-le-Pont, France.Sueur, J. (2001) Audiospectrographical analysis of cicada sound

production: a catalogue (Hemiptera: Cicadidae). Deutsche

Entomologische Zeitschrift, 48, 33–51.Sueur, J. (2002) Eco-ethologie de la Communication Sonore des

Cigales: le Modele Tibicina Amyot, 1847 (Hemiptera, Cicadidae,

Tibicininae). PhD Thesis, Ecole Pratique des Hautes Etudes,

Paris.Sueur, J. & Aubin, T. (2002a) Acoustic communication in the

Palaearctic red cicada Tibicina haematodes: chorus organisation,

calling song structure, and signal recognition. Canadian Journal

of Zoology, 80, 126–136.Sueur, J. & Aubin, T. (2002b) The decoding process in the acoustic

communication of the Palaearctic red cicada (Cicadomorpha,

Cicadidae, Tibicina haematodes): how males reply to synthetic

calling songs. Eleventh International Auchenorrhyncha Congress,

Postdam/Berlin, Germany, p. 37.Sueur, J. & Puissant, S. (2002) Spatial and ecological isolation in

cicadas: first data from Tibicina (Hemiptera: Cicadoidea) in

France. European Journal of Entomology, 99, 477–484.Sueur, J. & Puissant, S. (2003) Analysis of sound behaviour leads

to new synonymy in Mediterranean cicadas (Hemiptera,

Cicadidae, Tibicina). Deutsche Entomologische Zeitschrift, 50,

121–127.



Sueur, J., Puissant, S. & Pillet, J.-M. (2003) An Eastern

Mediterranean cicada in the West: first record of Tibicina

steveni (Krynicki, 1837) in Switzerland and France (Hemiptera,

Cicadidae, Tibicininae). Revue Francaise d’Entomologie, 25, in

press.Villet, M. (1995) Intraspecific variability in SMRS signals: some

causes and implications in acoustic signaling systems. Speciation



University Press, Baltimore, MD.Walker, T.J. (1974) Character displacement and acoustic insects.

American Zoologist, 14, 1137–1150.Wiley, R.H. & Richards, D.G. (1978) Physical constraint on

acoustic communication in the atmosphere: implications for the

evolution of animal vocalizations. Behavioral Ecology and

Sociobiology, 3, 69–94.Young, D. (1972) Neuromuscular mechanisms of sound produc-

tion in Australian cicadas. Journal of Comparative Physiology,

79, 343–362.

Young, D. (1980) The calling song of the bladder cicada,

Cystosoma saundersii: a computer analysis. Journal of Experi-

mental Biology, 88, 407–411.Young, D. & Bennet-Clark, H.C. (1995) The role of the tymbal in

cicada sound production. Journal of Experimental Biology, 198,

1001–1019.Young, D. & Josephson, R.K. (1983a) Mechanism of sound-

production and muscle contraction kinetics in cicadas. Journal of

Comparative Physiology A, 152, 183–195.Young, D. & Josephson, R.K. (1983b) Pure-tone songs in cicadas

with special reference to the genus Magicicada. Journal of

Comparative Physiology A, 152, 197–207.

Accepted 24 April 2003



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