Short Communication |
Corresponding author: Dávid Selnekovič ( david.selnekovic@uniba.sk ) Academic editor: Cheng-Bin Wang
© 2024 Dávid Selnekovič, Ján Kodada, Nilay Gülperçin, Serdar Tezcan, Enrico Ruzzier.
This is an open access article distributed under the terms of the Creative Commons Attribution License (CC BY 4.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.
Citation:
Selnekovič D, Kodada J, Gülperçin N, Tezcan S, Ruzzier E (2024) Morphological and molecular characterisation of Mordellistena peloponnesensis Batten, 1980 (Coleoptera, Mordellidae), with first records from Italy and Turkey. ZooKeys 1214: 105-117. https://doi.org/10.3897/zookeys.1214.133348
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Mordellistena peloponnesensis Batten, 1980, previously known from Cyprus and Greece, is reported from Italy and Turkey for the first time. The species is redescribed based on type specimens and additional material from its entire known distributional range. Eighteen DNA barcoding sequences of M. peloponnesensis from Greece, Cyprus, and Italy were generated, and genetic variability across the sampling localities was examined. Three mitochondrial haplotypes were detected within M. peloponnesensis. Specimens from mainland Italy share the same haplotype as those from Rhodes and Cyprus, whereas Sardinian specimens exhibit a distinct haplotype. The third haplotype is represented by one specimen from Cyprus. The DNA barcoding sequences of M. peloponnesensis were compared with those of the morphologically allied M. gemellata Schilsky, 1898, and M. pyrenaea Ermisch, 1966, to reveal the phylogenetic relationships between the species.
Distribution, DNA barcoding, haplotype, Mediterranean, morphology, tumbling flower beetles
Mordellistena A. Costa, 1854, is the largest genus within Mordellidae Latreille, 1802, with more than 150 species known to occur in Europe (
Our records significantly expand the known distribution of M. peloponnesensis, representing the northernmost and westernmost occurrences of the species. The study of extensive material, including type series, allowed us to provide a redescription of the species and a differential diagnosis to separate it from the morphologically allied species of the M. gemellata species group. We also explored the genetic variability of M. peloponnesensis across the sampled localities, showing that individuals from the Italian mainland share the same mitochondrial haplotype with individuals from Rhodes and Cyprus, whereas the Sardinian specimens exhibit a distinct, well-separated haplotype. The COI barcoding sequences generated during this study represent the first available DNA sequences for the species.
For this study, 108 individuals of M. peloponnesensis from Cyprus, Greece, Italy, and Turkey were examined, including 18 paratypes. Material is deposited in the following institutions and private collections: Finnish Museum of Natural History, Helsinki, Finland (
Specimens of M. peloponnesensis were compared with the type series of other species that are morphologically associated with the M. gemellata group: M. algeriensis Ermisch, 1966 (SNSD), M. aureotomentosa Ermisch, 1977 (SNSD), M. elbrusicola Ermisch, 1969 (SNSD), M. gemellata Schilsky, 1899 (
Individuals used for DNA isolation were killed in 96.3% ethanol and stored at -20 °C. DNA isolation and amplification procedures followed those provided by
Specimens were observed under an M205 C stereomicroscope (Leica, Wetzlar, Germany) with magnification up to 120× and diffused LED light (6400 K). Photographs of habitus were taken with an EOS 5D Mark II camera (Canon, Tokyo, Japan) attached to an Axio Zoom.V16 stereoscope (Zeiss, Oberkochen, Germany); photographs of genitalia were taken with an Axio Imager.M2 microscope (Zeiss, Oberkochen, Germany). The images were stacked using Zerene Stacker v.1.4 software (https://zerenesystems.com/cms/stacker) and edited in Adobe Photoshop CC (https://www.adobe.com/products/photoshop.html) and DxO Photolab 5 (https://www.dxo.com/dxo-photolab/). Measurements were made with an ocular micrometre in the M205 C stereomicroscope and are given in the text as the range, followed by the arithmetic mean and standard deviation enclosed in parentheses. The measured characters are abbreviated in the text as follows:
EL elytral length from scutellar apex to elytral apices along suture;
EW maximum elytral width;
HL head length from anterior clypeal margin to occipital carina along midline;
HW maximum head width;
LPrL maximum left paramere length;
PL pronotal length along midline;
PW maximum pronotal width;
PygL maximum pygidial length;
RPrL maximum right paramere length;
TL combination of head, pronotal and elytral lengths
Nucleotide sequences were trimmed and assembled into contigs using ChromasPro v.2.1.10 (Technelysium Pty Ltd, South Brisbane, Queensland, Australia). The final alignment was performed manually in Mesquite v.3.81 (
All voucher specimens used in this study, along with respective BOLD Sample and Process IDs and GenBank accession numbers, are listed in the linked data table (Suppl. material
Mordellistena peloponnesensis Batten, 1980: 42–44. Type locality: “Skála, Pelopónnesos, Greece”.
Mordellistena (s. str.) peloponnesensis:
Paratypes.
Greece • 3 males; Lakonia, Skala env. [same as holotype]; 9 Jul. 1978; R. Batten leg.; NBCL, RMNH.INS.1485999 [genitalia illustrated], RMNH.INS.1486000, RMNH.INS.1486001 • 4 males; Arkadhia, Tripolis env.; 5 Jul. 1977; R. Batten leg.; NBCL, RMNH.INS.1485986, RMNH.INS.1485987, RMNH.INS.1485988, RMNH.INS.1485989 • 1 male; Messinia, 2 km N. of Kardamili; 6 Jul. 1977; R. Batten leg.; NBCL, RMNH.INS.1485985 • 2 males; Messinia, Pilos env.; 8 Jul. 1978; R. Batten leg.; NBCL, RMNH.INS.1485996, RMNH.INS.1485997 • 1 male; Korinthia, Neméa env.; 10 Jul. 1978; R. Batten leg.; NBCL, RMNH.INS.1485990 • 1 male; same data as preceding;
Cyprus • 2 males, 3 females; Choirokoitia; 34.795833°N, 33.337500°E; alt. 186 m; 15 Jul. 2009; M. Mantič leg.; forest steppe, on flowers; MMCZ • 2 males; Pegeia, Coral Bay; 34.858333°N, 32.364167°E; alt. 9 m; 21 May 2017; M. Mantič leg.; forest steppe by sea, on flowers; MMCZ • 1 male, 6 females; Foinikaria, Germasogeia Reservoir; 34.755278°N, 33.093333°E; alt. 68 m; 27 Apr. 2018; D. Selnekovič leg.; secondary grassland, on Daucus; DSBS, DSBS15, DSBS16 • 1 male, 1 female; Kouklia env.; 34.666474°N, 32.628931°E; alt. 34 m; 9 May 2022; D. Selnekovič leg.; xeric grasslands on slopes, on Ferula; DSBS, DSBS609, DSBS610 • 1 male; Potamiou env.; 34.818289°N, 32.797439°E; alt. 677 m; 10 May 2022; D. Selnekovič leg.; road verge, orchards, on Daucus and Tordylium; DSBS, DSBS611 • 1 female; Pissouri; 34.653149°N, 32.715005°E; alt. 29 m; 12 May 2022; D. Selnekovič leg.; crop fields, slopes with xeric vegetation, on Ferula and Tordylium; DSBS • 3 males, 4 females; Avdimou env.; 34.675610°N, 32.758667°E; alt. 33 m; 13 May 2022; D. Selnekovič leg.; road verge, crop fields, on Daucus and Ferula; DSBS • 2 females; Kyrenia, Bellapais; 13 Jul 1939; H. Lindberg leg.;
Mordellistena peloponnesensis is characterised by antennomeres 1–4 being narrower than the following ones and by the presence of two short lateral ctenidia on the posterior tibia that are perpendicular to the dorsal edge of the tibia. This combination of characters is unique to the M. gemellata species group as defined by
The species most closely resembling M. peloponnesensis in terms of size, body shape, elytral length and width, and pubescence color are M. algeriensis Ermisch, 1966 (Algeria, Italy, and Tunisia), M. gemellata Schilsky, 1898 (Portugal, Spain), and M. pyrenaea Ermisch, 1966 (France, Spain). These species can be differentiated from M. peloponnesensis by distinct paramere shapes [see
Measurements (in mm; ♂♂ N = 13, ♀♀ N = 10): TL: ♂♂ 3.26–4.52 (3.91 ± 0.45), ♀♀ 3.98–4.87 (4.41 ± 0.32); HL: ♂♂ 0.61–0.80 (0.72 ± 0.07), ♀♀ 0.72–0.85 (0.79 ± 0.05); HW: ♂♂ 0.65–0.84 (0.76 ± 0.07), ♀♀ 0.72–0.94 (0.84 ± 0.07); PL: ♂♂ 0.75–1.06 (0.91 ± 0.12), ♀♀ 0.88–1.19 (1.04 ± 0.10); PW: ♂♂ 0.73–1.06 (0.91 ± 0.12), ♀♀ 0.86–1.17 (1.05 ± 0.09); EL: ♂♂ 1.91–2.68 (2.28 ± 0.27), ♀♀ 2.35–2.82 (2.58 ± 0.17); EW: ♂♂ 0.77–1.14 (0.95 ± 0.12), ♀♀ 0.97–1.25 (1.12 ± 0.09); RPrL: 0.25–0.32 (0.30–0.02); LPrL: 0.31–0.41 (0.36 ± 0.03).
Body elongate (Fig.
Head approximately as long as wide, HW/HL ratio: ♂♂ 1.02–1.12 (1.07 ± 0.03), ♀♀ 0.95–1.11 (1.07 ± 0.04), moderately convex dorsally, with highest point behind middle of eye length (lateral aspect); occipital carina rounded; integument weakly microreticulate, with small round setiferous punctures. Eyes broadly oval, approx. 1.4× longer than wide; posteriorly reaching to occipital margin; finely faceted; interfacetal setae longer than facet diameter. Anterior clypeal edge weakly convex. Labrum transverse, densely setose, anterior margin weakly convex. Antenna weakly serrate (Fig.
Pronotum approximately as long as wide, PL/PW ratio: ♂♂ 0.93–1.09 (1.00 ± 0.04), ♀♀ 0.96–1.02 (1.00 ± 0.02), widest before middle, strongly convex; surface microreticulate, densely covered with lanceolate setae, punctures larger than those on head; anterior edge convex in middle, anterior angles broadly rounded; lateral carinae strongly sinuate in lateral aspect; posterior edge sinuate, posterior angles rectangular in lateral aspect. Scutellar shield triangular, densely setose. Elytra moderately long and narrow, EL/EW ratio: ♂♂ 2.23–2.52 (2.40 ± 0.08), ♀♀ 2.18–2.42 (2.30 ± 0.07); apices separately rounded; surface microreticulate, densely covered with decumbent lanceolate setae, punctures coarser than those on pronotum. Hindwing fully developed. Metanepisternum approx. 4× longer than maximum width, narrowed posteriorly. Protarsomeres cylindrical, each little narrower than preceding one; protarsomere 1 as long as two following tarsomeres combined; penultimate protarsomere truncate distally, with apical edge weakly concave; each protarsal claw with three denticles; male protarsus with longer and thicker seta on ventral surface than female one. Mesotibia approx. 0.8× as long as mesotarsus. Mesotarsomeres cylindrical, each little narrower than preceding one; first mesotarsomere as long as two following tarsomeres combined. Metatibia with short subapical ctenidium and two lateral ctenidia nearly perpendicular to dorsal tibial edge, proximal ctenidium located around mid-length of tibia, distal one at around third quarter; outer terminal spur ca. 0.5× as long as inner one. Metatarsomere 1 with three ctenidia; metatarsomere 2 with two ctenidia; metatarsomeres 3 and 4 without lateral ctenidia.
Abdominal ventrite 5 with narrowly rounded apical edge. Pygidium long, conical, bent ventrad, narrowly truncate at apex, approx. 0.5× as long as elytra. Male sternite VIII with sinuate posterior edge, postero-lateral angles and medial portion moderately produced, with long trichoid sensilla (Fig.
Females on average slightly larger than males. Males somewhat slenderer than females. Second maxillary palpomere with longer setae in males than in females. Terminal maxillary palpomere slightly narrower in females. Male protibia bears several elongate setae in proximal half, female protibia uniformly setose. Male protarsomeres with numerous thick setae ventrally.
Eighteen sequences of the COI gene barcoding region were generated and submitted to BOLD (www.boldsystems.org) and GenBank (www.ncbi.nlm.nih.gov/genbank/). Details on voucher specimens as well as accession numbers are listed in the Suppl. material
Cyprus, Greece (mainland, Crete, Rhodes), Italy (southern part of the Italian peninsula, Sardinia, Sicily), Turkey (Fig.
A results of maximum likelihood analysis of 658 bp COI fragment; support values are given as: bootstrap values / aBayes test values B differences between haplotypes of Mordellistena peloponnesensis Batten, 1980. Codons are marked as the nucleotide positions within 658 bp COI fragment C TCS haplotype network based on eighteen sequences of 658 bp COI fragment in M. peloponnesensis; colours represent sampling localities; vertical lines represent number of substitutions D distribution of M. peloponnesensis; left: entire range, right: details on Greece, Turkey, and Cyprus. Localities of DNA samples are marked with coloured dots matching those in the haplotype network; localities without DNA samples are marked with black dots.
Five individuals of M. peloponnesensis from Sardinia were sampled in June 2021 from the inflorescences of Daucus carota L. (Apiaceae) in ruderal vegetation separating a parking lot from a small olive orchard in the urban area of Sassari. Three individuals from Vieste were collected on the inflorescences of D. carota in ruderal vegetation along a road. Five individuals from the vicinity of Naples were sampled in July 2023 on the inflorescences of D. carota in a ruderal habitat along a footpath on Camaldoli Hill. This area featured young secondary forest, orchards, and dry grassland communities. The following species co-occur with M. peloponnesensis in Italy: M. episternalis Mulsant, 1856, M. minima A. Costa, 1854, M. pseudorhenana Ermisch, 1977, M. purpurascens A. Costa, 1854, M. tarsata Mulsant, 1856, Mediimorda bipunctata (Germar, 1827), Variimorda basalis (A. Costa, 1854), and Stenalia sp. In similar ruderal habitats along roadsides, secondary grasslands, and olive orchards, specimens of M. peloponnesensis were also collected on the islands of Rhodes (Greece) and Cyprus.
We generated 18 sequences of the COI barcoding region of M. peloponnesensis from individuals originating from Cyprus, Greece (Rhodes) and Italy (mainland Italy and Sardinia). We recognised three haplotypes within the species: H1 shared by 12 individuals from Cyprus, Italy (Naples), and Greece (Rhodes); H2 represented by a single individual from Cyprus; and H3 shared by five individuals from Italy (Sardinia) (Fig.
We compared the sequences of M. peloponnesensis with those of the morphologically allied M. pyrenaea (four individuals from Spain) and M. gemellata (one individual from Spain). The divergence between M. peloponnesensis and M. gemellata is 7.94–8.09%, and between M. peloponnesensis and M. pyrenaea it is 3.34–4.00%. The intra-species divergence within M. peloponnesensis reaches a maximum value of 0.61%. Maximum likelihood analysis revealed a separate clade for each species (Fig.
Mordellistena peloponnesensis was described by Batten in 1980 based on specimens from various localities in southern mainland Greece and Peloponnese. Since its description, no further specific data have been published, except for its occurrence in Cyprus, as noted in the Catalogue of Palaearctic Coleoptera (
DNA sequences of M. peloponnesensis are currently available from the vicinity of Naples and Sardinia in Italy, from the island of Rhodes in Greece, and Cyprus. Analysis has shown that individuals from mainland Italy share the same haplotype as individuals from Rhodes and Cyprus. In contrast, individuals from Sardinia possess a distinct haplotype, separated by three nucleotide substitutions. Phylogenetic analysis suggested that the most closely related species is M. pyrenaea, known from Spain and France, with an interspecific divergence of 3.34–4.00%. The analysis did not include other morphologically similar species (M. algeriensis, M. aureotomentosa, M. maroccana, M. elbrusicola Ermisch, 1969, M. microgemellata Ermisch, 1965) for which DNA sequences were not available.
A parallel outcome from the examination of the type material is the identification of two syntypes of M. gemellata from Greece (Attica and Parnassus) as M. peloponnesensis. Likewise, two specimens from Kyrenia, Cyprus, originally identified and reported by
We would like to thank Oscar Vorst (NBCL), Győző Szél (
The authors have declared that no competing interests exist.
No ethical statement was reported.
This study was financially supported by the Slovak Research and Development Agency under project no. APVV-19-0076 and the Scientific and Technological Research Council of Turkey (TÜBİTAK), Grant 107K234. Additional support was provided by the SYNTHESYS+ Project (http://www.synthesys.info/) which is financed by European Community Research Infrastructure Action under the H2020 Integrating Activities Programme, Project number 823827. Enrico Ruzzier acknowledges the support of NBFC to the University of Roma Tre, Department of Science, funded by the Italian Ministry of University and Research, PNRR, “Missione 4 Componente 2, ‘Dalla ricerca all’impresa’, Investimento 1.4”, Project CN00000033.
Conceptualization: DS. Data curation: NG, ER, ST, DS. Formal analysis: DS. Funding acquisition: DS, ER. Validation: JK. Visualization: DS. Writing – original draft: DS. Writing – review and editing: ST, NG, JK, ER.
Dávid Selnekovič https://orcid.org/0000-0002-9228-1174
Ján Kodada https://orcid.org/0000-0002-1355-4323
Nilay Gülperçin https://orcid.org/0000-0002-9309-6528
Serdar Tezcan https://orcid.org/0000-0003-1980-9291
Enrico Ruzzier https://orcid.org/0000-0003-1020-1247
All of the data that support the findings of this study are available in the main text or Supplementary Information.
Aligned sequences of COI barcoding region
Data type: fas
Explanation note: Aligned sequences of 658 bp region of COI used in the present study.
List of DNA voucher specimens and sequence accession numbers
Data type: xlsx
Explanation note: Linked table containing data on DNA vouchers used in the present study.