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A Review of the Genus Mecistorhabdia (Lepidoptera: Erebidae: Arctiinae: Syntomini) with a Description of a New Species from the Central African Republic

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A Review of the Genus Mecistorhabdia (Lepidoptera: Erebidae: Arctiinae: Syntomini) with a Description of a New Species from the Central African Republic EUROPEAN JOURNAL OF ENTOMOLOGYENTOMOLOGY ISSN (online): 1802-8829 Eur. J. Entomol. 117: 139–148, 2020 http://www.eje.cz doi: 10.14411/eje.2020.016 ORIGINAL ARTICLE A review of the genus Mecistorhabdia (Lepidoptera: Erebidae: Arctiinae: Syntomini) with a description of a new species from the Central African Republic NIKOLAI IGNATEV 1, 2, BEATA GRZYWACZ 3 and ŁUKASZ PRZYBYŁOWICZ 3 1 Institute of Entomology, Biology Centre CAS, Branisovska 31, 370 05 Ceske Budejovice, Czech Republic, and Faculty of Sciences, University of South Bohemia, Ceske Budejovice, Czech Republic; e-mail: [email protected] 2 Museum Witt, Tengstraße 33, 80796 Munich, Germany 3 Institute of Systematics and Evolution of Animals, Polish Academy of Sciences, Sławkowska 17, 31-016 Krakow, Poland; e-mails: [email protected], [email protected] Key words. Lepidoptera, Erebidae, Arctiinae, Syntomini, Mecistorhabdia, new species, Afrotropical region, systematics, COI Abstract. The genus Mecistorhabdia Kiriakoff, 1953 is reviewed and one new species M. sulaki sp. n. from the Central African Republic and Cameroon (Afrotropical region) described. The holotype female of the type species M. haematoessa (Holland, 1893) is matched with a conspecifi c male using a molecular technique. M. burgessi Kiriakoff, 1957 is removed from the synonymy as it is considered to be a valid species. Habitus and male genitalia of all three species are illustrated along with the female genitalia of M. haematoessa. A key for identifi cation of species based on the structure of male genitalia and a map of their distribution are provided. The results of a study of the genetic diversity of the three species of Mecistorhabdia, based on cytochrome c oxidase subunit I gene, are presented. Relationships between species are briefl y discussed. ZooBank Article Registration: http://zoobank.org/urn:lsid:zoobank.org:pub:350BED21-ED91-4276-90E9-8AF1D2B2CCBC INTRODUCTION genus Mecistorhabdia. The diagnosis of the new genus For a long time the thyretines were thought to be a sepa- was based on signifi cant differences in the newly described rate monophyletic lineage and treated as a separate family, male genitalia in comparison with the genus Elsa (now the Thyretidae (Kiriakoff, 1949). Following morphologi- Rhipidarctia Kiriakoff, 1953) with which Mecistorhab- cal and molecular studies of the entire Noctuoidea it was dia was compared in the original description. He provided downgraded to tribal rank by Jacobson & Weller (2002) the description of male genitalia supplemented by rather and fi nally treated as a subtribe Thyretina of Syntomini schematic drawings, but did not describe the female geni- (Zahiri et al., 2011). Recently a comprehensive molecu- talia. Later Kiriakoff (1957) described one additional spe- lar study revealed that the Thyretina is not monophyletic cies, M. burgessi, from Uganda based on a single male. (Przybyłowicz et al., 2019). As a result, it was formally This new taxon was subsequently synonymized with M. synonymized with Syntomini. The group was revised by haematoessa by Przybyłowicz & Kühne (2008). These au- Przybyłowicz (2009), but since then very few papers have thors wrongly treated the small differences in colouration been published. Among these, Durante & Potenza (2014) and male genitalia structures as intraspecifi c variability. listed the species of the genus Balacra Hampson, 1914 re- The same misinterpretation was repeated by Przybyłowicz corded in Gabon and Durante & Zangrilli (2016) revised (2009) in his catalogue of Thyretini. the subgenus Daphaenisca Kiriakoff, 1953 and described The study of type specimens of both taxa and additional three new species. material from adjacent territories has made it possible to Until recently the genus Mecistorhabdia included only a study this genus in more detail and as a consequence to single valid species, M. haematoessa, which was originally describe another new taxon. Using molecular techniques, placed by Holland (1893) in the genus Metarctia Walker, we managed to associate correctly the female holotype of 1855. This species was described based on a single female M. haematoessa with known males and prove the morpho- without description of its reproductive organs. Kiriakoff logical distinctiveness of taxa by using molecular and mor- (1953) revised the status of this taxon and proposed a new phological approach. We verify the distribution of species Final formatted article © Institute of Entomology, Biology Centre, Czech Academy of Sciences, České Budějovice. An Open Access article distributed under the Creative Commons (CC-BY) license (http://creativecommons.org/licenses/by/4.0/). 139 Ignatev et al., Eur. J. Entomol. 117: 139–148, 2020 doi: 10.14411/eje.2020.016 and provide high-quality illustrations of the reproductive sary with the old female holotype of M. haematoessa (specimen organs enabling an easier separation of species. No. 18.4.6), which is more than 100 years older than the other material. If one of these reactions was successful, an effort was MATERIALS AND METHODS made to obtain a barcode compliant record (> 650 bp) by amplify- ing shorter regions of COI. PCR were performed in 25 μl reaction In this study we follow the systematic account of Przybyłowicz volume using 2 × Phanta Max Master Mix (Vazyme Biotech Co., et al. (2019) for the higher taxonomic levels. Ltd, China), 10 mM dNTP Mix, 10 μM of each primer and 4 μl of Specimens of the genus Mecistorhabdia were collected in the genomic DNA. The PCR profi le consisted of an initial dena- Uganda (July 2011 by Harald Sulak), Central African Republic turation step at 94°C for 3 min, followed by 36 cycles at 94°C for (September 2000 by local collector) and Republic of the Congo 1 min, 48°C for 1 min, and 72°C for 2 min and a fi nal extension (January to March 1997 by Viktor Siniaev and Vladimir Murzin) step of 7 min at 72°C. Successful PCR products were purifi ed and stored in the MWM collection. Additional material from the using the GeneMATRIX PCR/DNA Clean-Up Purifi cation kit RMCA and ISEAPAS collections was used for comparison. (EURx, Poland, following the standard protocol) and sequenced The abdomens were removed and placed in cold 5% KOH so- in both directions. All sequences were deposited in GenBank lution overnight after which the abdominal cuticle was opened under the accession numbers MN441758 and MN441760– by a right lateral cut to the genitalia, which were removed and MN441768 (Table 1). The sequence of Mecistorhabdia burgessi stained in Evans Blue. The aedeagus was pulled out caudally. Im- from the GenBank database was also included in the analysis ages of genitalia were taken using a Leica DFC450 camera. (Accession Number MK158553). Arctia caja (Linnaeus, 1758) Forewing length was measured from the base to the apex of the and Lithosia quadra (Linnaeus, 1758) in the tribe Arctiini and wings, parallel to the wing costa. Forewing width was measured Lithosiini, respectively (both in subfamily Arctiinae), were cho- perpendicularly from the costa to the tornus. sen as outgroup for analysis and the sequences were also taken Morphological terms for genitalia are those used by Klots from the GenBank database (Accession Numbers: HM873709 (1970). and KJ547685, respectively). Abbreviations: FW – forewing; GP – genitalia slide; HW – The obtained nucleotide sequences were edited and assembled hindwing; CMNH – Carnegie Museum of Natural History, Pitts- into contigs in CodonCode Aligner (CodonCode Corporation). burgh, USA; ISEAPAS – Institute of Systematics and Evolution Sequences were also checked for protein-coding frame shifts of Animals, Polish Academy of Sciences, Kraków, Poland; MWM to detect pseudogenes using MEGA X (Kumar et al., 2018) and – Museum Witt, Munich, Germany; NECUJ – Nature Education compared with sequences from GenBank using a BLAST search. Centre of Jagiellonian University, Kraków, Poland; NHMUK – Best-fi t models for Bayesian (BI) and maximum likelihood (ML) Natural History Museum, London, UK; RBINS – Royal Belgian analyses were calculated in MrModeltest (Nylander, 2004) using Institute of Natural Sciences, Brussels, Belgium; RMCA – Royal the Akaike information criterion (AIC). ML analyses were car- Museum for Central Africa, Tervuren, Belgium; ZSM – Zoologis- ried out in IQTree (Nguyen et al., 2015). Bootstrap support was che Staatssammlung des Bayerischen Staates, Munich, Germany. calculated using 1000 replicates. BI analyses were performed in Molecular analysis MrBayes (Ronquist & Huelsenbeck, 2003) with four independ- Total genomic DNA was extracted from leg muscle tissue of ent runs, each having three heated and one cold chain. Analyses dried specimens or specimens preserved in pure ethanol using the were run for 6 million generations with trees sampled every 1000 GeneMATRIX Bio-Trace DNA Purifi cation kit (EURx, Poland, generations. The fi rst 25% of each run was discarded as burn-in. following the standard protocol for tissue). The cytochrome c Convergence among the runs was assessed using Tracer (Ram- oxidase subunit I gene (COI) was amplifi ed by polymerase chain baut et al., 2018). All trees were visualized using FigTree with a reaction (PCR) using the primers LCO1490 (5’-GGT CAA CAA midpoint rooting option (Rambaut & Drummond, 2012). ATC ATA AAG ATA TTG G-3’) and HCO2198 (5’-TAA ACT Pairwise sequence divergences were calculated separately for TCA GGG TGA CCA AAA AAT CA-3’) (Folmer et al., 1994). In intraspecifi c as well as interspecifi c comparisons with MEGA X case of failure in sequencing with these primers, additional PCR using Kimura’s two parameter (K2P) distance model. Species reactions were carried out to recover amplicons using primers delimitation method Automatic Barcode Gap Discovery, ABGD ZBJ-ArtF1c (5’-AGA TAT TGG AAC WTT ATA TTT TAT TTT (Puillandre et al., 2012), is an automatic procedure that consid- TGG-3’) and ZBJ-ArtR2c (5’-WAC TAA TCA ATT WCC AAA ers the sequences as hypothetical species based on the barcoding TCC TCC-3’) (Zeale et al., 2011). This combination was neces- gap. The model fi rst calculates a range of prior intraspecifi c diver- Table 1.
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