PL-ISSN0015-5497(print),ISSN1734-9168(online) FoliaBiologica(Kraków),vol.61(2013),No1-2 Ó InstituteofSystematicsandEvolutionofAnimals,PAS,Kraków,2013 doi:10.3409/fb61_1-2.125 ReassessmentoftheSystematicPositionof Orthocomotis DOGNIN (Lepidoptera:Tortricidae) BasedonMolecularDatawithDescriptionofNewSpeciesofEuliini JózefRAZOWSKI,SebastianTARCZ, JanuszWOJTUSIAK,andVolkerPELZ Accepted November 22, 2012 RAZOWSKI J., TARCZ S., WOJTUSIAK J., PELZ V. 2013. Reassessment of the systematic position of Orthocomotis DOGNIN (Lepidoptera: Tortricidae) based on molecular data with description of new species of Euliini. Folia Biologica (Kraków) 61: 125-134. The application of molecular analyses for resolving taxonomic problems in the family Torticidae (Lepidoptera) is still uncommon. The majority of papers concern the assessment of population variability of economically important species; reports on the systematic positions of Neotropical Tortricidae taxa are rare. The Neotropical genus Orthocomotis was classifiedinitiallyasamemberofthetribeEuliini.Then, basedongenitalmorphology,itwas moved to the tribe Polyorthini. A comparison of homologous 606 bp fragments of the COI mitochondrial gene revealed that Orthocomotis should be transfered back into the tribe Euliini. Based on an analysis of phylogenetic relationshipsthestudiedgeneraof Euliini form a monophyletic cluster, clearly separated from tribe Polyorthini in which they were temporarilyincluded.Moreover,inthecurrentpaperwedescribetwonew speciesofthetribe Euliini: Galomecalpa lesta RAZOWSKI &PELZ, sp. n., Gauruncus ischyros RAZOWSKI & PELZ, sp. n. Key words: Tortricidae, Orthocomotis, molecular phylogeny, mitochondrial COI. Józef RAZOWSKI, Department of Invertebrate Zoology, Institute of Systematics and Evolution of Animals, Polish Academy of Sciences, 31-016 Kraków, S³awkowska 17, Poland. E-mail: [email protected] Sebastian TARCZ, Department of Experimental Zoology, Institute of Systematics and Evolu- tion of Animals, Polish Academy of Sciences, 31-016 Kraków, S³awkowska 17, Poland. E-mail: [email protected] Janusz WOJTUSIAK, Zoological Museum, Jagiellonian University, Ingardena 6, 30-060 Kraków, Poland. Volker PELZ, Bonnenweg 3, D-503809 Ruppichteroth, Germany. The Tortricidae, a family of Lepidoptera with LING 2011) species complex. The earliest analyses global occurrence, consists of about 10000 de- were based on allozyme comparisons, for example scribed species (BROWN 2005) of which a large the degree of genetic differentiation of the larch number do not have clearly defined taxonomic po- budmoth Zeiraphera diniana was studied using sitions.Ataxonomicsystembasedonmorphologi- 24allozymeloci(EMELIANOV et al. 1995). Subse- cal characters of members of this family has been quently, methods based on PCR began to play improved for more than 150 years, but, as in most a more significant role. Introgression between two other insect groups, is far from definite. Compara- closely related species of the genus Choristoneura tive analysis of genome fragments provides an op- was confirmed by RAPD markers (DEVERNO et al. portunity for taxonomic progress and determination 1998). Another approach, AFLP markers, was suc- of the relationships among various taxa. cessfully applied for reconstruction of the phylo- Information on the applications of molecular genetic position of Cydia pomonella (THALER et al. markers in Torticidae for the identification of the 2008). systematic position of particular taxa is rather Sequencing homologous DNA fragments pro- sparse. Most evaluate the population structure of vides an opportunity for the parallel analysis of a economically important species, for example Tor- largernumberoffeaturesthaninthemethodsmen- trix viridana (SCHROEDER &DEGEN 2008), Tor- tioned above. However studies of intra-specific re- tricidae from South Africa (TIMM et al. 2010) or lationships among Torticidae taxa are mostly the Choristoneura fumiferana (LUMLEY &SPER- based on comparisons of mitochondrial genome 126 J.RAZOWSKI etal. fragments, especially the cytochrome oxidase motis requires further study. The purpose of the gene (COI). For example, the phylogenetic rela- present study is to elucidate this issue. tionships of Argyrotaenia franciscana were deter- In the present paper we compared several spe- mined by analysis of a mitochondrial DNA cies of Orthocomotis forming a compact grouping fragment containing the genes COI and COII and used four representative species. We com- (2300 bp) (LANDRY et al. 1999). A similar analy- pared Orthocomotis with six species of Poly- sis using the COI segment was carried out by com- orthini and Palaearctic Eulia ministrana, the paring sequences obtained from closely related type-species of the genus Eulia, and five represen- species of the genus Archips (KRUSE & SPERLING, tatives of the Neotropical Euliini. In the Palaearc- 2002). A 940 bp COI fragment was applied to dis- tic there is another species (Pseudargyrotoza criminate between two forms of Adoxophyes conwagana FABRICIUS, 1775) but there are some orana feeding on different food plants in the objections if it belongs to Euliini (HORAK 1999). Adoxophyes species complex (LEE et al. 2005). In the Neotropics there are numerous species of A combination of morphological and molecular Euliini which are included in the tribe on the basis data was useful for verification of the systematic of presence of the pedal scent organ. position of the European Tortricini (RAZOWSKI et al. Becauseoftheuncertainpositionof Orthocomo- 2010) and for resolving uncertain relationships be- tis, we proposed a preliminary molecular approach tween tribes Bactrini and Endotheniini (RAZOWSKI asatoolforresolvingthistaxonomicproblem.The &TARCZ 2012). However, such analyses have not present paper is the first comparative molecular been applied to Neotropical Torticidae genera. study of representatives of the studied species. For the first time genus Orthocomotis was de- scribedforoneNeotropicalspecies. Orthocomotis didn’t have a tribal placement and was treated as a Material and Methods member of Tortricinae. CLARKE (1955) was the first to revise the genus and included in it 29 spe- cies (12 previously known and 17 newly described Material species). His supposition that Orthocomotis is The examined specimens were collected by a genus was confirmed by further studies (RAZOWSKI Janusz WOJTUSIAK and Volker PELZ in Ecuador et al. 2007). and are preserved in the collection of the Zoologi- Then RAZOWSKI (1982) transferred Orthoco- cal Museum of the Jagiellonian University, motis to Polyorthini, a tribe of the subfamily Chli- Kraków (MZUJ) and the Volker PELZ collection. danotinae based on the following synapomorphy: Due to problems associated with obtaining good a minutely bristled dorsal portion of the anellus quality DNA suitable for molecular analysis, COI situated immediately above the aedeagus, con- sequences of Eulia ministrana were taken from nected with the aedeagus and transtilla. GenBank. Representatives of the tribe Olethreu- tini i.e. Apotomis inudana, Apotomis sauciana, AccordingtoPOWELL (1986) this genus belongs Olethreutes arcuellus were used as outgroups. in Euliini. However, POWELL (1986) based his A list of examined taxa arranged alphabetically is analysis only on some synplesiomorphies of the presented in Table 1. genitalia. Finally BROWN (1989) transferred Orthocomo- Molecular methods tis and its closely related genus Paracomotis RAZOWSKI, 1982 to Schoenotenini based on the DNA was extracted only from two hind legs of chaetosema situated on the vertex of the head. He dry specimens because some of them were mu- also confirmed a previous hypothesis (DIAKON- seum material (it was impossible to use other parts OFF 1974) that the most important character of of the bodies – e.g. the entire tagmata) and pre- Polyorthini is the presence of the outer slit of the served in 70% alcohol. The examined specimens valva and its associated corema (a scalepencil were not older than 10 years and were first identi- fromtheterminalpartoftheabdomenconcealedin fied by comparison of the genitalia. Specimens a slit). However, there are many closely related over ten years of age usually gave insufficient re- species of Chlidanotinae with a secondarily re- sults. The best results were obtained from 1-3 year duced abdominal scent organ. old individuals preserved in 70% alcohol. Recently, there was a consensus that the dis- Genomic DNA was isolated without protocol cussed genus belongs to Euliini (HORAK 1999; modification using the NucleoSpin Tissue Kit HORAK &BROWN 1991; RAZOWSKI 2008) al- (Macherey-Nagel, Germany). To elute purified though RAZOWSKI and BECKER (1999) suggested DNA we applied 100 Fl of Elution Buffer (EB) onto that the true phylogenetic placement of Orthoco- the silica membrane. To amplify a fragment of the Table 1 Tortricidaespeciesusedinthisstudy. Newlydescribedtaxonsaremarkedgrey.ThreespeciesfromthetribeOlethreutiniwereusedasanoutgroup No. DNAVoucher Tribe Genus Species Origin Authors COI acc RAZOWSKI and WOJTU- 1. TORT023 Euliini Orthocomotis parandina Ecuador SIAK, 2010 JX144962 RAZOWSKI and WOJTU- 2. TORT055 Euliini Orthocomotis marmorobrunnea Ecuador SIAK, 2006 JX144963 RAZOWSKI and WOJTU- 3. TORT058 Euliini Orthocomotis oxapampae Ecuador SIAK, 2010 JX144964 4. TORT056 Euliini Orthocomotis golondrina Ecuador RAZOWSKI et al., 2007 JX144965 RAZOWSKI and PELZ, Reassessmentof 5. VP40 Euliini Dimorphopalpa lyonsae Ecuador 2007 JX144966 6. VP53 Euliini Seticosta sp. Ecuador – JX144967 RAZOWSKI and BECKER, 7. VP02