Tettigettalna josei (Boulard, 1982)

Simoes, Paula C., Nunes, Vera L., Mendes, Raquel, Seabra, Sofia G., Paulo, Octavio S. & Quartau, Jose A., 2014, Tettigettalnajosei (Boulard, 1982) (Hemiptera: Cicadoidea): first record in Spain, with notes on the distribution, genetic variation and behaviour of the species, Biodiversity Data Journal 2, pp. 1045-1045 : 1045

Tettigettalna josei (Boulard, 1982)

Materials

Type status: Other material. Occurrence: recordedBy: Bruno Novais; individualCount: 1; sex: male; Location: country: Portugal; stateProvince: Algarve; verbatimLocality: Budens; verbatimLatitude: 37°04'45.2"N; verbatimLongitude: 8°50'11.6"W; Event: samplingProtocol: Sweep net capture; eventDate: 2011-07-27; Record Level: collectionID: Tjo119; institutionCode: FCUL; collectionCode: Entomology_PCS

Type status: Other material. Occurrence: recordedBy: Bruno Novais; individualCount: 1; sex: male; Location: country: Portugal; stateProvince: Algarve; verbatimLocality: Budens; verbatimLatitude: 37°04'22.9"N; verbatimLongitude: 8°48'43.9"W; Event: samplingProtocol: Sweep net capture; eventDate: 2011-07-27; Record Level: collectionID: Tjo122; institutionCode: FCUL; collectionCode: Entomology_PCS

Type status: Other material. Occurrence: recordedBy: Bruno Novais; individualCount: 1; sex: male; Location: country: Portugal; stateProvince: Algarve; verbatimLocality: Porches; verbatimLatitude: 37°08'09.4"N; verbatimLongitude: 8°23'04.2"W; Event: samplingProtocol: Sweep net capture; Acoustic recording; eventDate: 2011-07-26; Record Level: collectionID: Tjo113; institutionCode: FCUL; collectionCode: Entomology_PCS

Type status: Other material. Occurrence: recordedBy: Bruno Novais; individualCount: 1; sex: male; Location: country: Portugal; stateProvince: Algarve; verbatimLocality: Vale Judeu; verbatimLatitude: 37°07'39.8"N; verbatimLongitude: 8°05'36.1"W; Event: samplingProtocol: Sweep net capture; Acoustic recording; eventDate: 2011-07-12; Record Level: collectionID: Tjo66; institutionCode: FCUL; collectionCode: Entomology_PCS

Type status: Other material. Occurrence: recordedBy: Vera Nunes; individualCount: 1; sex: male; Location: country: Portugal; stateProvince: Algarve; verbatimLocality: Quinta do Lago; verbatimLatitude: 37°03'35.2"N; verbatimLongitude: 8°01'16.3"W; Event: samplingProtocol: Sweep net capture; Acoustic recording; eventDate: 2012-08-01; Record Level: collectionID: Tjo309; institutionCode: FCUL; collectionCode: Entomology_PCS

Type status: Other material. Occurrence: recordedBy: Vera Nunes; individualCount: 1; sex: male; Location: country: Portugal; stateProvince: Algarve; verbatimLocality: Quinta do Lago; verbatimLatitude: 37°03'35.2"N; verbatimLongitude: 8°01'16.3"W; Event: samplingProtocol: Sweep net capture; Acoustic recording; eventDate: 2012-08-08; Record Level: collectionID: Tjo355; institutionCode: FCUL; collectionCode: Entomology_PCS

Type status: Other material. Occurrence: recordedBy: Vera Nunes; individualCount: 1; sex: male; Location: country: Portugal; stateProvince: Algarve; verbatimLocality: Quinta do Lago; verbatimLatitude: 37°03'35.2"N; verbatimLongitude: 8°01'16.3"W; Event: samplingProtocol: Sweep net capture; Acoustic recording; eventDate: 2012-08-09; Record Level: collectionID: Tjo362; institutionCode: FCUL; collectionCode: Entomology_PCS

Type status: Other material. Occurrence: recordedBy: Bruno Novais; individualCount: 1; sex: male; Location: country: Portugal; stateProvince: Algarve; verbatimLocality: S. Brás de Alportel; verbatimLatitude: 37°08'14.8"N; verbatimLongitude: 7°50'52.4"W; Event: samplingProtocol: Sweep net capture; Acoustic recording; eventDate: 2011-08-04; Record Level: collectionID: Tjo145; institutionCode: FCUL; collectionCode: Entomology_PCS

Type status: Other material. Occurrence: recordedBy: Vera Nunes; individualCount: 1; sex: male; Location: country: Portugal; stateProvince: Algarve; verbatimLocality: Moncarapacho; verbatimLatitude: 37°04'41.3"N; verbatimLongitude: 7°49'16.6"W; Event: samplingProtocol: Sweep net capture; Acoustic recording; eventDate: 2011-08-03; Record Level: collectionID: Tjo141; institutionCode: FCUL; collectionCode: Entomology_PCS

Type status: Other material. Occurrence: recordedBy: Bruno Novais; individualCount: 1; sex: male; Location: country: Portugal; stateProvince: Algarve; verbatimLocality: Moncarapacho; verbatimLatitude: 37°04'41.3"N; verbatimLongitude: 7°49'16.6"W; Event: samplingProtocol: Sweep net capture; Acoustic recording; eventDate: 2011-08-10; Record Level: collectionID: Tjo154; institutionCode: FCUL; collectionCode: Entomology_PCS

Type status: Other material. Occurrence: recordedBy: Bruno Novais; individualCount: 1; sex: male; Location: country: Portugal; stateProvince: Algarve; verbatimLocality: Tavira; verbatimLatitude: 37°08'02.0"N; verbatimLongitude: 7°38'04.2"W; Event: samplingProtocol: Sweep net capture; eventDate: 2011-08-11; Record Level: collectionID: Tjo159; institutionCode: FCUL; collectionCode: Entomology_PCS

Type status: Other material. Occurrence: recordedBy: Vera Nunes; individualCount: 1; sex: male; Location: country: Portugal; stateProvince: Algarve; verbatimLocality: Castro Marim; verbatimLatitude: 37°11'10.9"N; verbatimLongitude: 7°29'02.1"W; Event: samplingProtocol: Sweep net capture; Acoustic recording; eventDate: 2011-08-02; Record Level: collectionID: Tjo137; institutionCode: FCUL; collectionCode: Entomology_PCS

Type status: Other material. Occurrence: recordedBy: Vera Nunes; individualCount: 1; sex: male; Location: country: España; stateProvince: Huelva; verbatimLocality: Cartaya; verbatimLatitude: 37°15'38.4"N; verbatimLongitude: 7°07'43.5"W; Event: samplingProtocol: Sweep net capture; Acoustic recording; eventDate: 2013-07-17; Record Level: collectionID: Tjo3557; institutionCode: FCUL; collectionCode: Entomology_PCS

Type status: Other material. Occurrence: recordedBy: Vera Nunes; individualCount: 1; sex: male; Location: country: España; stateProvince: Huelva; verbatimLocality: Cartaya; verbatimLatitude: 37°15'38.4"N; verbatimLongitude: 7°07'43.5"W; Event: samplingProtocol: Sweep net capture; Acoustic recording; eventDate: 2013-07-18; Record Level: collectionID: Tjo3562; institutionCode: FCUL; collectionCode: Entomology_PCS

Type status: Other material. Occurrence: recordedBy: Vera Nunes; individualCount: 1; sex: male; Location: country: España; stateProvince: Huelva; verbatimLocality: Cartaya; verbatimLatitude: 37°15'38.4"N; verbatimLongitude: 7°07'43.5"W; Event: samplingProtocol: Sweep net capture; Acoustic recording; eventDate: 2013-07-18; Record Level: collectionID: Tjo3566; institutionCode: FCUL; collectionCode: Entomology_PCS

Type status: Other material. Occurrence: recordedBy: Vera Nunes; individualCount: 1; sex: male; Location: country: España; stateProvince: Huelva; verbatimLocality: Cartaya; verbatimLatitude: 37°14'03.7"N; verbatimLongitude: 7°03'56.8"W; Event: samplingProtocol: Sweep net capture; eventDate: 2013-07-18; Record Level: collectionID: Tjo3577; institutionCode: FCUL; collectionCode: Entomology_PCS

Description

Taxonomic identification

Specimens collected in Spain were identified based on acoustic, morphological and genetic analysis. Acoustic analysis of the calling song of three males (Fig. 2, Table 1, Suppl. material 1) confirmed that their song’s profile is in agreement with previous descriptions of Tettigettalna josei acoustics ( Fonseca 1991). The calling song is composed of the repetition of a long sequence of phrases. Each phrase includes two parts, Part I with a long sequence of echemes separated by very short intervals and Part II shorter than Part I, at the end of the phrase, with echemes produced continuously and ever decreasing inter-echeme interval duration. Tettigettalna josei specimens have a broad spectrum near 9 - 22.5 kHz with maximum energy around 17 kHz. For time domain variables, results obtained for Spanish specimens indicated an echeme duration ranging from 0.002 to 0.009s, with an average value of 0.004s. For the echeme period we found a range of 0.002 to 0.058s with an average of 0.015s. Morphological measurements of specimens collected in Spain are presented in Table 2. We found an average of 19.21 mm for total body length and 16.22 mm for hindwing length. These values are in general agreement with the ones previously reported for Tettigettalna josei ( Boulard 1982). Sequences of cytochrome c oxidase I (COI) obtained from the three specimens collected in Spain (GenBank: KF977503-KF977505) were identical to the ones reported in Nunes et al. (2014) for Tettigettalna josei (GenBank: KC807267-KC807274) thus confirming species identification.

Genetic variation

We combined COI sequences from Nunes et al. (2014) and from this study in a dataset composed by 23 male specimens of Tettigettalna josei from several locations across its distribution (Table 4) and constructed a minimum spanning network (Fig. 5a). The dataset included eight variable sites, resulting in nine haplotypes that differ from each other by single mutations. All mutations result in synonymous changes in the protein. Haplotypes H1 and H6 were the ones found in higher frequencies, but they were detected only in a few sampled locations, indicating that the distribution of some haplotypes tends to be localized (Fig. 5b).

Distribution

Calling songs from males of Tettigettalna josei were consistently heard in the region of Algarve, from the west Atlantic coast until the surroundings of the easternmost town, Vila Real de S. António (Fig. 3). A set of new locations and georeferenced coordinates of occurrence of Tettigettalna josei are listed in Table 3, but they should not be considered as an exhaustive list. Tettigettalna josei is quite widespread in Algarve, reaching high densities (> 10 singing males) in open habitats covered with low vegetation (small bushes and dry grass) and well exposed to sunlight (Fig. 4). Consequently, their numbers were low in cultivated tree groves where herbaceous vegetation was removed but were easily found in uncultivated fields or small patches of marginal vegetation by roads, in the periphery of villages and in secondary dunes and cliffs near the sea. Tettigettalna josei was found in sympatry with two other species belonging to the same genus, Tettigettalna argentata and Tettigettalna mariae (Table 3). Males of Tettigettalna josei were also found singing on trees, but they usually sing at heights below three meters, unlike their congenerics Tettigettalna argentata or Tettigettalna mariae , which often sing perched on high pine trees.

In July 2013, Tettigettalna josei was also found in small numbers in Cartaya (Huelva, Spain), which extends its known distribution to Spain. This same area was visited the year before (see Simões et al. 2013), but Tettigettalna josei was not found then. We surveyed other provinces of Andalusia in July of 2012 and 2013, but Tettigettalna josei was not found so far in any other areas besides Huelva.

Biology

Reproductive behaviour

During our fieldwork we observed the copulatory mating behaviour of Tettigettalna josei . We had witnessed a few ongoing copulations in other Tettigettalna species ( Tettigettalna argentata and Tettigettalna helianthemi ) but had never seen how the process is initiated. One male of Tettigettalna josei was first noticed in a branch tip of a small stone pine ( Pinus pinea ) while singing and its unusual behaviour caught our attention. The male was moving frantically up and down or circling while singing. We noticed a female standing still in the same branch but on the opposite side of the male, thus out of sight of the male. The female was standing still and produced wing-flicks at regular and short intervals (see Boulard (2006) for a review on cicadas wing-flicking). After a few wing-flicks the male eventually moved into the females’ direction, turned around the branch and made body contact with her. The male immediately mounted on the female’s dorsum and initiated a sideway copulation (Fig. 6). The couple stood quietly and in silence for about three minutes, being the female’s front legs firmly attached to the branch. When the couple become apart, the female remained in the branch while the male took off a minute later to a nearby branch and resumed his calling song. A female of this species was seen laying their eggs on a Fennel stem ( Foeniculum vulgare ) at about 1 m from the soil. Cicadas of genus Tettigettalna are particularly vulnerable during copulation and oviposition because they tend to resist fleeing away if threatened.

Taxon discussion

The present data clarify the currently known distribution range of Tettigettalna josei in the south of the Iberian Peninsula, encompassing the region of Algarve (Portugal) and the province of Huelva (Spain). Morphological, acoustic and genetic analyses confirmed the identity of specimens collected in Cartaya (Huelva) as belonging to Tettigettalna josei .

Previous records on the distribution of this small cicada species were sparse and limited to Portugal ( Sueur et al. 2004). Hence the species has been considered until now as endemic to Portugal. The new record of Tettigettalna josei in Spain in July 2013 refutes this status and the species must now be added to the list of the Spanish cicadas and, therefore, should be considered as an Iberian endemism.

The region of Huelva was previously surveyed in August 2012 with the detection of Tettigettalna mariae ( Simões et al. 2013) but not of Tettigettalna josei . This could be explained by the low densities of Tettigettalna josei found in Spain, since only about five singing males were detected in the region in 2013. Additionally, we have noted that the emergence peak of Tettigettalna josei might be slightly earlier (June-July) as compared to other Tettigettalna species ( Tettigettalna argentata and Tettigettalna mariae : mostly July). Consequently, the number of singing males of Tettigettalna josei should decline earlier, in mid-August, justifying why we might have missed Tettigettalna josei in Spain during our surveys in mid-August 2012.

Data obtained so far indicates that populations of Tettigettalna josei are acoustic, morphological and genetically homogeneous throughout the distribution range of the species. The genetic analysis of COI gene showed no evidence of population structure. Haplotypes differ by single mutations from each other and form a star-like haplotype network. In spite of this, some differences in the distribution and frequency of each COI haplotype seem to reflect the expected trend for low dispersal in these cicadas. As demonstrated before in a species of genus Cicada ( Simões and Quartau 2007), emerged specimens usually experience limited dispersal during the mating season, remaining near the emergence site, where the singing males may form choruses.

The distribution of Tettigettalna josei overlaps with an area under severe human pressure. The coastline of Algarve has been intensively exploited for beach tourism and golf. However, land management associated with tourism facilities seem to have less impact on the persistence of Tettigettalna josei than the perturbation caused by farming practices that deplete the shrub and grass cover of the soil (e.g. plowing, harvesting or intensive grazing). Fortunately for cicadas, these practices are not severely intense in the coastal region of Algarve and crops are usually small sized and patchy, allowing populations of Tettigettalna josei to persist all over the region. In contrast, monocultures such as olive and stone pine woods in the Spanish region of Andalusia occupy extensive areas and are regularly maintained to restrain the growth of shrub-like vegetation under the trees. These differences in land management might help to explain the small effective numbers of Tettigettalna josei detected so far in Spain, and favours instead the prevalence of Tettigettalna species that are frequently found on trees, such as Tettigettalna mariae or Tettigettalna aneabi .