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Corresponding author: Lech Karpiński ( lechkarpinski@gmail.com ) Academic editor: Steffen Pauls
© 2021 Lech Karpiński, Wojciech T. Szczepański, Radosław Plewa, Lech Kruszelnicki, Katarzyna Koszela, Jacek Hilszczanski.
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.
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This paper sheds the first light on the phylogeny of the Central Asian genus Turanium Baeckmann, 1922. By applying an integrative taxonomy approach, we revealed and described a new species from Kyrgyzstan—Turanium losi Karpiński, Plewa & Hilszczański sp. nov. Distinguishing characters from closely related Turanium pilosum (Reitter, 1891) are presented and their ecological associations are discussed. The key characters, including the male terminalia, were examined by means of scanning electron microscopy. High-quality stacked photographs of the habitus of the specimens are presented for both species and their geographical distributions are mapped. While the new species shows stable morphological characters that allow its differentiation from T. pilosum and the COI genetic distance between them is approx. 3%, the different species delimitation methods gave discordant results. Although the new species remained unrecognized for so long, it seems that these cerambycids are common in the region and both can be considered potentially invasive as they are apparently highly polyphagous. It has also been documented that they occur sympatrically in Kyrgyzstan. Both the Bayesian and maximum likelihood analyses of COI sequences confirmed the monophyly of the genus Turanium with strong support (PP 1 and BS 90, respectively). Moreover, the recently revealed polyphyly of the tribe Callidiini was supported by our analyses and, consequently, the discussion on the establishment of a new tribe Ropalopini is raised. This study further corroborates the effectiveness of DNA barcoding as a tool in detecting new species and provides some of the first sequences for Central Asian cerambycids, which remain almost completely unknown in terms of molecular studies.
BOLD, DNA barcoding, Central Asia, longhorned beetles, SEM, taxonomy
The genus Turanium Baeckmann, 1922 is a small group of the family Cerambycidae, subfamily Cerambycinae that is distributed in Central Asia—from northeastern Iran (Bojnord) to central and southeastern Kazakhstan (Dzungarian Alatau) and western part of Kyrgyzstan, including almost the entire territory of Tajikistan (
DNA barcoding is a technique that involves sequencing of 658 bp from the 5’ end of the mitochondrial cytochrome oxidase subunit I gene (
During the routine barcoding of longhorned beetles distributed in Central Asia, which remain almost completely unknown regarding their molecular sequences, we noticed that the specimens of ‘Turanium pilosum’ that were sampled in Tajikistan and Kyrgyzstan formed two well-defined clades. Hence, this study aimed at investigating (morphology and species delimitation methods) the existence of a possible new taxon and at better understanding the taxonomic position of this group by revealing results of the first molecular-based (COI) phylogeny of the genus Turanium and providing the sequences for further studies. Taking advantage of the opportunity, we also decided to examine the phylogenetic position of the morphologically closest genera of the tribe in trying to test the recently questioned (
This study is based on the examination of more than 80 specimens of the genus Turanium from the territory of Kyrgyzstan, Tajikistan, Kazakhstan and Uzbekistan. The following acronyms for institutional and private collections are used in the text:
CJH Collection of Jacek Hilszczański, Sękocin Stary, Poland
CKL Collection of Krzysztof Łoś, Łomianki, Poland
CLK Collection of Lech Kruszelnicki, Siemianowice Śląskie, Poland
CRP Collection of Radosław Plewa, Sękocin Stary, Poland
MIZ Museum and Institute of Zoology, Polish Academy of Sciences, Poland
Other abbreviations used: BI, Bayesian inference; BL, body length; BS, maximum likelihood bootstrap values; HT, holotype; ML, maximum likelihood inference; PP, Bayesian posterior probability; PT, paratype.
The individuals that were used for the detailed morphological and molecular analyses were collected by the authors during the entomological expeditions to Tajikistan (2014), Kazakhstan (2017, 2018) and Kyrgyzstan (2018, 2019). Some of the localities that were posted on professional websites (www.cerambyx.uochb.cz; www.zin.ru/animalia/coleoptera) were taken into consideration and presented only when combined with photographs, enabling accurate identification. After publication, the holotype will be deposited in the collection of MIZ.
The beetles were examined using an Olympus SZH10 Stereo Microscope at 7–140× magnification, a PROLAB MSZ Stereo Microscope at 7–90× magnifications, and a Hitachi S-3400N Scanning Electron Microscope. To examine the sclerotized parts of the male terminalia, the specimens were relaxed in distilled water for 12–24 h at room temperature. Then, the genitalia and last abdominal segment were separated from the other abdominal structures using pins or forceps, without removing the rest of the abdomen. Separated genitalia were put into 15% KOH solution at room temperature for some 24 h.
The scanning electron microscope (SEM) images were taken using a Hitachi S-3400N SEM at MIZ. Photographs of the habitus were taken with a Canon EOS 50D digital camera equipped with a Canon 100 mm f/2.8 USM Macro lens. The images that were produced were stacked, aligned, and combined using Helicon Focus (www.heliconsoft.com) and Zerene Stacker (www.zerenesystems.com) software. Photographs of the habitats were taken with a Canon EOS 600D and a Nikon Coolpix AW110 cameras. All plates were prepared using Adobe Photoshop CS5 and GIMP v2.10.14. The distribution of the species was illustrated in Quantum GIS (QGIS) v3.6.0 ‘Noosa’ (
DNA barcoding, the analysis of a standardised segment from the 5’ end of the mitochondrial cytochrome c oxidase subunit I (COI) gene, was performed on a representative selection of species (four of eight including members of both subgenera; six specimens) of the targeted group: T. pilosum (Kyrgyzstan), T. losi Karpiński, Plewa & Hilszczański sp. nov., T. scabrum (Kazakhstan), and T. johannis (Kyrgyzstan) (Table S1). Additionally, we barcoded specimens of two extremely rare Central Asian species from the most closely related genus Ropalopus Mulsant, 1839: Ropalopus nadari Pic, 1894 and Ropalopus mali Holzschuh, 1993, which belong to the same tribe (Callidiini) and occur in the same habitats. Sequencing of some additional specimens that were collected, killed and preserved under the same conditions was unsuccessful, however, each species tested has obtained at least one sequence of a satisfactory length. We chose this gene because it has proven very informative in our previous studies (e.g.
The individuals that were reared from the inhabited material collected in the field (T. losi Karpiński, Plewa & Hilszczański sp. nov., Kyrgyzstan) were preserved in 96% ethanol, which was subsequently replaced in order to avoid diluting the alcohol. For the remaining species, only dried specimens (collected between 2014 and 2019; killed with ethyl acetate) were utilised. The specimens were processed for DNA barcoding in 2020. The right mid femur was cut open on both ends to expose the muscle tissue and then partly crushed with forceps and placed in a sealed well that contained two drops of 95% ethanol on a standard 96-well microplate, which was used for the tissue submission.
All of the laboratory work for extracting, purifying, amplifying and sequencing the DNA was performed at the ‘Canadian Centre for DNA Barcode’ (CCDB, http://www.ccdb.ca), University of Guelph, Ontario, Canada, following the standard protocol (
Due to the quality of the material, only eight specimens were successfully sequenced for a 619–658 bp long DNA barcoding fragment. The sequences were submitted to GenBank under the accession numbers OK073067–OK073074 (Table S1).
The obtained sequences and additional relevant information such as the specimen images, primers, gel images and trace files were uploaded to the ‘Barcode of Life Database’ (=BOLD, http://www.boldsystems.org) in the public online dataset ‘Turanium Central Asia LK’ (DS-LKCCAT; DOI: dx.doi.org/10.5883/DS-LKCCAT). All of the voucher specimens reported herein are part of the BOLD project ‘LECHK Cerambycidae Central Asia L. Karpinski’ and were deposited in the LK’s Cerambycid DNA-grade specimen bank at MIZ and in the collection of Forest Research Institute, Poland.
Phylogenetic trees were reconstructed using both Bayesian inference (BI) and maximum likelihood (ML) methods. The obtained trees were also used to infer possible species delimitations. In addition to the barcoded specimens, sequences of the following species of Callidiini (11 species of 6 genera) and the most closely related tribes (16 species of 6 genera in 3 tribes) were obtained from GenBank and BOLD as closest related outgroup: Callidiini—Callidium aeneum [KM286341.1], Callidium coriaceum [KU918865.1], Callidium violaceum [KU914026.1], Oupyrrhidium cinnabarinum [KY683721.1], Phymatodes pusillus [KM285884.1], Phymatodes rufipes [HQ953905.1], Phymatodes testaceus [HQ954560.1], Pyrrhidium sanguineum [KU918701.1], Ropalopus femoratus [KM446501.1; KJ964459.1; KM286267], Ropalopus sanguinicollis [COLAT043-08; CERLF390-08], Semanotus japonicus [LC492880.1]; Cerambycini—Cerambyx cerdo [KM285966.1], Cerambyx scopoli [KJ962934.1], Cerambyx miles [KM286032.1]; Clytini—Chlorophorus annularis [MK689190.1], Chlorophorus diadema [KC135923.1], Chlorophorus figuratus [JF889542.1], Chlorophorus sartor [KM449257.1], Chlorophorus signaticollis [FJ559042.1], Chlorophorus varius [KM286012.1], Clytus arietis [JF889522.1], Clytus lama [KU918981.1], Plagionotus arcuatus [JF889541.1], Plagionotus detritus [KM442177.1], Xylotrechus rusticus [KM286086.1], Xylotrechus stebbingi [MN182963.1], and Hylotrupini—Hylotrupes bajulus [MH020456.1]. Prionus coriarius [MH020283.1], belonging to the subfamily Prioninae was chosen as the most distantly related outgroup. The resulting topology was visualized in FigTree v1.4 (
Sequences of COI were aligned in Geneious v9.1.7 (Biomatters Ltd, Auckland, New Zealand, 2005) using the MAFFT plugin v1.3.6, based on MAFFT (
Bayesian analysis was performed using MrBayes v3.2.7a (
Maximum likelihood (ML) analysis was run with RAxML v8.2.12 (
We considered maximum likelihood bootstrap values (BS) 90–100 as strong, 75–89 as moderate, and 50–74 as weak; Bayesian posterior probability from MrBayes (PP) 0.95–1 as strong, 0.85–0.94 as moderate, and 0.70–0.84 as weak support. Nodes with BS < 50 and PP < 0.70 were considered to be unsupported.
Distance-based and tree-based species delimitation methods were performed to investigate species boundaries of T. losi Karpiński, Plewa & Hilszczański sp. nov. In order to receive a robust estimate of its entities, three approaches were used: Assemble Species by Automatic Partitioning (ASAP;
The ASAP distance-based analyses were run on the ‘ASAP web’ server (https://bioinfo.mnhn.fr/abi/public/asap) using Kimura 2-Parameter as a substitution model for matrices of genetic distances. Only partitions showing the lowest asap-score were considered.
The best-score ML tree from RAxML was used as input for PTP, with the most distantly related outgroup removed. ML inference was performed using the single-rate method and was run with mPTP v0.2.4 (
The tree from MrBayes was used for bPTP analysis which was run on the ‘bPTP server’ (species.h-its.org). Similarly to the previous analyses, the most distantly related outgroup was removed. MCMC chains were run for 500 thousand generations, and all other settings were left as default.
Initiated by the conspicuous divergence in COI sequences, detailed morphological studies of the specimens representing both sequenced populations from Tajikistan and Kyrgyzstan confirmed the existence of two closely related species. Subsequently, additional individuals representing populations from other localities in the region were investigated. The results of this study are presented as both SEM and stacked colour plates. The general habitus of the beetles is presented dorsally (Fig.
Habitus (dorsal view) of some representatives of the genera Turanium and Ropalopus. A–E Turanium losi Karpiński, Plewa & Hilszczański sp. nov.: male holotype (4 km N of Arkit, Kyrgyzstan), female (ibid), male (8 km N of Arkit, Kyrgyzstan), female (Kara-Alma, Kyrgyzstan), female (Torkent, Kyrgyzstan), respectively F–H Turanium pilosum: male (Takob, Tajikistan), female (ibid), male (Kara-Alma, Kyrgyzstan), respectively I, J Turanium scabrum: male (Kara-Alma, Kyrgyzstan), female (Karashota, Kazakhstan), respectively K, L Turanium johannis: male (Urumbash, Kyrgyzstan), female (ibid), respectively M Ropalopus nadari female (Takob, Tajikistan) N Ropalopus mali male (Karakul, Kyrgyzstan). Scale bar: 3 mm.
8 ♂♂ and 13 ♀♀. — Holotype: male (Figs
Comparison of particular body parts of two Turanium species. A–C, G–I, M Turanium losi Karpiński, Plewa & Hilszczański sp. nov.: A head of male holotype B pronotum of male holotype C pronotum of female (Kara-Alma, Kyrgyzstan) G elytra (apex) of male holotype H elytra (basal part) of male holotype I elytra (basal part) of female (Kara-Alma, Kyrgyzstan) M prosternal process of male (type locality) D–F, J–L, N, O Turanium pilosum: D head of male (Takob, Tajikistan) E pronotum of male (ibid) F pronotum of female (ibid) J elytra (apex) of male (ibid) K elytra (basal part) of male (ibid) L elytra (basal part) of female (ibid) N prosternal process of male (ibid) O prosternal process of female (ibid).
Morphology. Body relatively slender, strongly flattened dorsoventrally; BL in males: 10.5–14.0 mm (HT 12.0 mm), in females: 11.0–15.0 mm. Humeral width in males: 3.0–3.9 mm (HT 3.5 mm), in females: 3.0–4.2 mm. Integument black; legs and antennae black; elytra black (sometimes with slightly brownish areas on sides behind middle) with slight metallic luster. Pubescence of whole body made by sparse but distinct, long and usually erect whitish hairs and short black setae, being the longest on pronotum and anterior part of elytra, in its posterior part with many shorter and semi-decumbent hairs especially along epipleura; on ventral side distinct long whitish hairs, varying in intensity among specimens but usually denser on abdominal sternites; on femora semi-decumbent, relatively long and sparse whitish hairs; on tibiae and tarsi mostly replaced by denser blackish setae forming brush; on antennae very dense, relatively long, erect setae, especially rich on first five antennomeres and gradually disappearing towards last joint, on last antennomeres in form of single spike-like setae. Head broad, with coarse sculpture; frons with longitudinal furrow of variable depth between antennal tubercles; clypeus and labrum broad and well-pronounced; mandibles and palpi stout; eyes large, surrounding antennal tubercles; genae narrow, approx. 0.15–0.2 of eye width. Antennae thick and strong, relatively long, exceeding elytral apex by 1.5–2 last antennomeres in males, and clearly shorter, never reaching elytral apex in females; average length ratio of antennomeres in males: 1:0.25:1.25:1.0:1.08:1.08:1.13:1.0:0.93:0.83:1.13, in females: 1:0.25:1.2:0.95:1.0:0.95:0.95:0.86:0.8:0.7:0.9; antennomeres 3–10 with tooth of variable depth on outer side; in males antennomere 11 always clearly divided in approx. 2/3 of length, forming almost completely separated antennomere 12. Prothorax very wide and pronounced, distinctly narrower at the base, gradually widening towards upper edge, strongly flattened, with clearly rounded outer edges, in females less pronounced and slightly more oblong, also with clearly rounded outer edges; approx. 1.32 (HT 1.37) in males and 1.4 in females times as wide as long, approx. 3.65 (HT 3.6) in males and 4.15 in females times shorter than elytral length and 1.2 (HT 1.2) in males and 1.26 in females times narrower than elytra at humeri. Pronotum rather regularly (less regularly in males) and entirely punctate, in males usually with smooth area at middle near base; punctation rather uniform (more uniform in females), relatively sparse but clearly finer and denser at sides. Prosternum finely and sparsely punctate, distinctly creased longitudinally. Prosternal process short and very thin, barely exceeding middle of procoxae, slightly curved dorsally. Elytra moderately long, approx. 2.25 (HT 2.22) in males and 2.33 in females times as long as humeral width, gradually tapering towards apex in males and usually almost parallel-sided in females, in both sexes with single indentation on each elytron about 1/3 of anterior length; elytral sculpture composed mainly by regular, sparse, fine, shallow punctures, with coarse surface between them (Fig.
Variability of particular body parts in two Turanium species. A, B, F–H, K–M Turanium losi Karpiński, Plewa & Hilszczański sp. nov.: A head of male (type locality) B head of female (Kara-Alma, Kyrgyzstan) F pronotum of male (type locality) G pronotum of male (ibid) H pronotum of male (Kara-Alma, Kyrgyzstan) K elytra (basal part) of male (type locality) L elytra (basal part) of male (ibid) M elytra (basal part) of male (Kara-Alma, Kyrgyzstan) C–E, I, J, N, O Turanium pilosum: C head of male (Takob, Tajikistan) D head of male (Arkit, Kyrgyzstan) E head of female (Takob, Tajikistan) I pronotum of male (ibid) J pronotum of male (Arkit, Kyrgyzstan) N elytra (basal part) of male (Takob, Tajikistan) O elytra (basal part) of female (ibid).
Variability of particular body parts in two Turanium species. A, B, E, G, K, M Turanium losi Karpiński, Plewa & Hilszczański sp. nov.: A elytra (apex) of male (type locality) B elytra (apex) of female (Kara-Alma, Kyrgyzstan) E metafemur (ventral view) of male (type locality) G abdomen (ventral view) of male (ibid) K first two antennomeres (ventral view) of male (ibid) M tarsal pads of protarsomere 2 and 3 (ventral view) of male (ibid); C, D, F, H–J, L, N, O Turanium pilosum: C elytra (apex) of male (Takob, Tajikistan) D elytra (apex) of female (ibid) F metafemur (ventral view) of male (ibid) H abdomen (ventral view) of male (ibid) I abdomen (ventral view) of male (ibid) J abdomen (ventral view) of female (ibid) L first two antennomeres (ventral view) of male (ibid) N tarsal pads of protarsomere 2 and 3 (ventral view) of male (ibid) O tarsal pads of protarsomere 2 and 3 (ventral view) of female (ibid).
COI sequences of two individuals of the new species were uploaded to BOLD and GenBank under the accessions: LK0101/LK0114 and OK073067/ OK073074, respectively. The new taxon was registered under Barcode Index Number (BIN): BOLD:AEF9068.
Turanium losi Karpiński, Plewa & Hilszczański sp. nov. differs from its closest relative, T. pilosum (Figs
Lateral lobes of Turanium species. A, B, G Turanium losi Karpiński, Plewa & Hilszczański sp. nov.: male holotype, male (type locality), male (ibid), respectively C–F Turanium pilosum: male (Kara-Alma, Kyrgyzstan), male (Takob, Tajikistan), male (ibid), male (Arkit, Kyrgyzstan), respectively H Turanium scabrum male (Kara-Alma, Kyrgyzstan) I Turanium johannis male (Urumbash, Kyrgyzstan).
Summary of the intraspecific and mean interspecific COI genetic distances between the representatives of the genera Turanium and Ropalopus, estimated using maximum composite likelihood model. See Supplementary Table (S2) for the genetic distances between all analysed species and individuals. ‘n/a’ indicates species in which only a single specimen was sequenced.
Intraspecific variability [%] | Species | Mean interspecific distances [%] | |||||||
1 | 2 | 3 | 4 | 5 | 6 | 7 | |||
0.2 | Turanium pilosum | 1 | |||||||
0.0 | Turanium losi sp. nov. | 2 | 2.7 | ||||||
n/a | Turanium scabrum | 3 | 12.1 | 11.2 | |||||
n/a | Turanium johannis | 4 | 12.3 | 11.5 | 13.6 | ||||
n/a | Ropalopus nadari | 5 | 13.6 | 13.0 | 13.8 | 14.9 | |||
n/a | Ropalopus mali | 6 | 17.3 | 15.6 | 17.2 | 18.4 | 11.6 | ||
0.0 | Ropalopus sanguinicollis | 7 | 14.7 | 14.6 | 17.7 | 15.5 | 16.0 | 17.2 | |
0.0–0.6 | Ropalopus femoratus | 8 | 16.4 | 16.5 | 17.7 | 16.8 | 15.3 | 14.2 | 16.1 |
The new species can also easily be separated from T. rauschorum (habitus of male paratype presented in
Other species in the genus clearly differ in body proportions and colouration, the punctation of the pronotum and elytra, and other characters. The habitus of T. scabrum and T. johannis is presented in Fig.
Turanium losi Karpiński, Plewa & Hilszczański sp. nov. is known to occur in northwestern Kyrgyzstan and easternmost Uzbekistan (Fig.
Adults of the new species were collected from the first days of May to the third quarter of June, at altitudes between 1000 and 1900 m a.s.l. The earliest observation was done at the lowest altitude (May 6, at 994 m a.s.l.). The holotype and the main series of paratypes have been reared from larvae collected in mid-June at an altitude of 1500 m a.s.l. At the time, imagoes of T. pilosum (exclusively males) were collected in the same locality and despite careful investigation no adults of the new species were observed at this plot. At the higher altitude (1620 m a.s.l.), females of the new species were found on June 20, together with a single male of T. pilosum. In type locality in Kyrgyzstan the species is related to mountain deciduous open forests with a substantial share of Fraxinus L. (Oleaceae), Prunus L., and Malus L. trees (Rosaceae) (Fig.
The new species seems to be common in the region and its larvae are most likely wide polyphages as the adults that were reared from Kyrgyz Prunus L. and Malus L. were able to continue breeding on the wood of European hornbeam Carpinus betulus L. (Betulaceae). Moreover,
Based on own observations, the life cycle usually lasts two years, however in the laboratory rearing, a single male emerged from the wood material that was added already in Poland (II generation) after only eight months, which means that the cycle can be shortened in nature to one year under optimal conditions.
As T. losi Karpiński, Plewa & Hilszczański sp. nov. is rather common in Middle Asia, there were already some specimens imaged in the Internet or in scientific papers that present the new species but were clearly misidentified. One example could be a male of ‘T. johannis’ from Uzbekistan (www.cerambyx.uochb.cz; accessed on: 10.01.2021, the image has been replaced) and another a female of ‘Turanium rauschorum’ from Kyrgyzstan, close to the Kazakh border (www.zin.ru). Regarding the revision of the genus (
We are pleased to dedicate this species to a Polish entomologist, Krzysztof Łoś, our close friend and one of the main organizers of the 2017–2019 trips to Kazakhstan and Kyrgyzstan.
Two following partitions were found: COI1 + COI2 and COI3. For MrBayes SYM+I+G model was selected as the best supported for the first one and HKY+G for the second. The maximum likelihood (Fig.
Callidiini was recovered as polyphyletic, separated in a few clades, although generally with weak support value of each subgroup: Phymatodes + Pyrrhidium as a part of Clytini, and Callidium and Ropalopus + Turanium as separate (ML) or unresolved (BI) clades.
Maximum likelihood phylogenetic tree based on the COI sequences of the representatives of Turanium and Ropalopus and some closely related genera of Callidiini and other Cerambycinae tribes. Maximum likelihood bootstrap values >50 are shown. Clades within Callidiini were marked by coloured branches as follows: purple—Turanium, blue—Ropalopus, red—remaining genera. Additional interpretation for the trib labels on the right side: purple—Hylotrupini, blue—Cerambycini.
Bayesian phylogenetic tree based on the COI sequences reporting the results of the species delimitation analyses (Turanium + Ropalopus clade). Bayesian posterior probabilities >0.70 are shown. Vertical bars correspond to morphology (purple) and to the species delimitation results obtained by ASAP (red), bPTP (green), and PTP (blue) methods.
For additional verification, we also tested the putative new species using a few species delimitation methods. Different methods gave, however, discordant results, either confirming the species status of T. pilosum and T. losi Karpiński, Plewa & Hilszczański sp. nov., accepting them partly, or rejecting them as distinct taxa. Turanium losi Karpiński, Plewa & Hilszczański sp. nov., besides strong morphological support, was confirmed as a separate species in the ASAP method. On the contrary, the PTP method recognised both Turanium as a single taxon, while the bPTP method gave inconclusive results, supporting their divergence in 72% of analyzes (while the author of the method suggests treating results as sufficiently accurate with the support of approx. 0.90). It is worth mentioning, however, that two species of the genus Plagionotus Mulsant, 1842: Plagionotus arcuatus (Linnaeus, 1758) and Plagionotus detritus (Linnaeus, 1758) have also been revealed as a single species by this method, with very similar support (0.78) as for Turanium taxa. Similarly, in the PTP method, both Plagionotus species were revealed as a single taxon. There are no justified reasons not to consider P. arcuatus and P. detritus as two separate species. The COI genetic distance between them is approx. 3%, nearly the same as in case of T. pilosum and T. losi Karpiński, Plewa & Hilszczański sp. nov. (Table S2). We also investigated that the authors of the original paper (
The results from each method are presented on the Bayesian inference tree (Fig.
Callidiini is a cosmopolitan tribe comprising 30 genera (41 including subgenera) (Tavakilian and Chevillotte 2021). After exclusion of Callidium Fabricius, 1775 and Semanotus Mulsant, 1839, which in our study were separated from Turanium Baeckmann, 1922 and Ropalopus Mulsant, 1839 and clustered in
Besides the monophyly of the genus Turanium our analyses revealed the polyphyly of the tribe Callidiini. Although a COI-based phylogeny should not be overinterpreted, we decided to present and discuss some parts of our trees as very similar results were obtained by
Formation of analogous clades despite the use of different markers and different species sampling seems suggestive. As it was mentioned above, neither Ropalopus nor Turanium were analysed in
Since we present here the first sequences of Turanium and, next to one Palaearctic and one Nearctic species available in GenBank/BOLD, the only sequences of Ropalopus (sole for Asian species), these results seem quite novel and except herein widely referenced paper of
Turanium is an exclusive Central Asian genus that includes generally highly polyphagous species. For instance, according to
Another issue is the invasive potential of Turanium. At least half of the species are common beetles in Central Asia and their adults, considering individual populations, are present in nature almost throughout the entire growing season. Moreover, it was documented that larvae are also able to develop in dry wood (T. scabrum;
We would like to dedicate this work to late Dr. Ottó Merkl, who for many years was the senior curator, and recently also the director, of the Department of Zoology, Hungarian Natural History Museum in Budapest and who was the SYNTHESYS+ host of LKa while studying the type material of Turanium. We thank our colleagues: Krzysztof Łoś (the main organizer), Tomasz Jaworski, Grzegorz Tarwacki, and Yerlan Shaperov for their participation in the entomological expeditions to Kazakhstan and Kyrgyzstan in 2017–2019. We would also like to show our gratitude to Dagmara Żyła (Museum and Institute of Zoology, Polish Academy of Sciences, Poland) for her assistance in conducting phylogenetic analyses and shared experience in molecular work. Finally, we thank an anonymous referee and the subject editor Steffen Pauls, whose detailed comments helped to improve the manuscript.
Table S1
Data type: .pdf
Explanation note: List of barcoded Turanium and Ropalopus specimens, their GenBank accessions/BOLD sample IDs, with the collecting data.
Table S2
Data type: .pdf
Explanation note: Genetic divergence between the COI sequences (in %) estimated using the maximum composite likelihood model implemented in MEGA7.
Sequence alignment
Data type: .fas
Explanation note: Data file used for phylogenetic analyses.