A DNA barcode library of the Mediterranean species of Eumerus (Diptera: Syrphidae) 1 Chroni ,
*, 2 Djan ,
3 Vidaković ,
2 Vujić ,
Antonia Mihajla Dragana Obreht Ante Theodora 1 4 2 2 1 Petanidou , Llilja Šašić Zorić , Nevena Velićković , Nataša Kočiš Tubić , George Tataris , Snežana Radenković2 1 University of the Aegean, Department of Geography, University Hill, 81100, Mytilene, Greece 2 University of Novi Sad, Faculty of Sciences, Department of Biology and Ecology Serbia 3 University of British Columbia’s Department of Forest and Conservation Sciences, Faculty of Forestry, Vancouver, BC Canada 4 University of Novi Sad, BioSense Institute – Research Institute for Information Technologies in Biosystems, Serbia
[email protected]
Eumerus Meigen, 1822 (Diptera: Syrphidae) High species diversity: 256 species worldwide, 39 in the Mediterranean (including countries of the Balkan Peninsula, Chroni et al., unpubl.; Grković et al., unpubl.). New species are constantly being introduced (9 new species in the last two years). Presence of species complexes (E. minotaurus complex, Chroni et al., unpubl.). One new record (E. uncipes, Grković et al., unpubl.), one species in revision (E. alpinus, Grković et al., 2015), several endemics, two major monophyletic clades and seven taxon groups (Chroni et al., 2017).
Species delimitation is a challenge due to scarcity of an up-to-date European identification key. Employment of DNA barcodes only recently started, have led to inference of the phylogenetic relationships among species and have altered our way of perception; there are more species out there to discover and more issues to resolve.
Aim To construct a DNA barcode library of all Eumerus species occurring in the Mediterranean Region and four adjacent Balkan countries.
Material and Methods
Characteristics
Dataset
# sequences
345
1. 34 species (out of the 39 Mediterranean ones); 10 countries and 75 localities (Fig. 1). 2. 5’-end fragment of the mitochondrial COI gene (DNA barcode, Folmer et al., 1994). 3. A dataset of 345 Eumerus sequences and four outgroups: one sequence of Platynochaetus setosus and Megatrigon tabanoides, as well as two sequences of Merodon erivanicus. 4. Polymorphic sites, DNA polymorphism and basic molecular diversity indices were calculated using DnaSP 5.10.01 (Table 1). 5. Bayesian inference employed in MrBayes 3.2.6 in the Cipres Science Gateway, under the GTR + G substitution model (as suggested by BIC, employed in MEGA 6.06), partitioned data by codon (2 partitions: positions 1st+2nd; 3rd).
Length (bp)
617
Singleton variable sites
28
Parsimony informative sites
149
No haplotypes
125
Haplotype (gene) diversity, Hd
0.9653
Nucleotide diversity, Pi
0.0586
Table 1. Characteristics of the dataset; outgroups are not considered in the ‘# sequences’.
Results - Discussion All 34 species were successfully delimitated by the DNA barcode (Fig. 2).
Fig. 1. Distribution of the 34 Eumerus species occurring in the Mediterranean region. Each number corresponds to species: 1 – E. alpinus Rondani, 1857, 2 - E. amoenus Loew, 1848, 3 – E. argyropus Loew, 1848, 4 – E. armatus Ricarte & Rotheray, 2012, 5 – E. aurofinis Grković, Vujić & Radenković, 2015, 6 – E. basalis Loew, 1848, 7 – E. clavatus Becker, 1923, 8 – E. consimilis Šimić & Vujić, 1996, 9 – E. crassus Grković, Vujić & Radenković, 2015, 10 – E. emarginatus Loew, 1848, 11 – E. grandis Meigen, 1822, 12 – E. hungaricus Szilady, 1940, 13 – E. karyates Chroni, Grković & Vujić, sp. n., 14 – E. lucidus Loew, 1848, 15 – E. minotaurus Claussen & Lucas, 1988, 16 – E. montanum Grković, Radenković & Vujić sp. n., 17 – E. niveitibia Becker, 1921, 18 – E. nudus Loew, 1848, 19 – E. obliquus (Fabricius, 1805), 20 – E. ornatus Meigen, 1822, 21 – E. ovatus Loew, 1848, 22 – E. pannonicus Ricarte, Vujić & Radenković, 2016, 23 – E. phaeacus Chroni, Grković & Vujić, sp. n., 24 – E. pulchellus Loew, 1848, 25 – E. pusillus Loew, 1848, 26 – E. sinuatus Loew, 1855, 27 – E. sogdianus Stackelberg, 1952, 28 – E. strigatus (Fallén, 1817), 29 – E. sulcitibius Róndani, 1868, 30 – E. torsicus Grković & Vujić, 2015, 31 – E. tricolor (Fabricius), 1798, 32 – E. truncatus Rondani, 1868, 33 – E. uncipes Rondani, 1850, 34 – E. vestitus Bezzi, 1912.
Fig. 1. Sample locations and distribution of the 34 Eumerus species occurring in the Mediterranean region. Each number corresponds to a species: 1 – E. alpinus Rondani, 1857; 2 – E. amoenus Loew, 1848; 3 – E. argyropus Loew, 1848; 4 – E. armatus Ricarte & Rotheray, 2012; 5 – E. aurofinis Grković, Vujić & Radenković, 2015; 6 – E. basalis Loew, 1848; 7 – E. clavatus Becker, 1923; 8 – E. consimilis Šimić & Vujić, 1996; 9 – E. crassus Grković, Vujić & Radenković, 2015; 10 – E. emarginatus Loew, 1848; 11 – E. grandis Meigen, 1822; 12 – E. hungaricus Szilady, 1940; 13 – E. karyates Chroni, Grković & Vujić, sp. n.; 14 – E. lucidus Loew, 1848; 15 – E. minotaurus Claussen & Lucas, 1988; 16 – E. montanum Grković, Radenković & Vujić sp. n.; 17 – E. niveitibia Becker, 1921; 18 – E. nudus Loew, 1848; 19 – E. obliquus (Fabricius, 1805); 20 – E. ornatus Meigen, 1822; 21 – E. ovatus Loew, 1848; 22 – E. pannonicus Ricarte, Vujić & Radenković, 2016; 23 – E. phaeacus Chroni, Grković & Vujić, sp. n.; 24 – E. pulchellus Loew, 1848; 25 – E. pusillus Loew, 1848; 26 – E. sinuatus Loew, 1855; 27 – E. sogdianus Stackelberg, 1952; 28 – E. strigatus (Fallén, 1817); 29 – E. sulcitibius Róndani, 1868; 30 – E. torsicus Grković & Vujić, 2015; 31 – E. tricolor (Fabricius), 1798; 32 – E. truncatus Rondani, 1868; 33 – E. uncipes Rondani, 1850; and 34 – E. vestitus Bezzi, 1912.
Future implications (a) Taxonomic revision of the genus: How many species and endemics are really? Are there more species complexes? (b)Employment of more molecular markers and inclusion of more samples e.g. from the Western Mediterranean. (c) Phylogeographic and biogeographic studies: Which historical processes (paleoclimatic and paleogeologic events) and biogeographic processes (vicariance or dispersal) have led speciation within Eumerus? Fig. 2. Bayesian analysis for the 5’-end fragment of the COI gene, partitioned by codon; values indicate Bayesian posterior probability, and circles denote 100% posterior probability (only values > 50% are illustrated). Photos of the species: A. Chroni.
References Chroni, A., Grković, A., Ačanski, J., Vujić, A., Radenković, S., Veličković, N., Djan, Μ., Petanidou, T. (unpubl.) Is there more out there? Disentangling cryptic species complex and new species within Eumerus longicornis group (Diptera: Syrphidae) based on integrative taxonomy and Aegean palaeogeography. Chroni, A., Djan, M., Vidaković, D.O., Petanidou, T.., Vujić, A. (2017) Molecular species delimitation in the genus Eumerus (Diptera: Syrphidae). Bulletin of Entomological Research, 107(1), pp. 126–138, http://dx.doi.org/10.1017/S0007485316000729 Folmer, O., Black, M., Hoeh, W., Lutz, R., Vrijenhoek, R. 1994. DNA primers for amplification of mitochondrial cytochrome c oxidase subunit I from diverse metazoan invertebrates. Mol. Mar. Biol. Biotechnol. 3(5), 294–299. Grković, A., Vujić, A., Chroni, A., van Steenis, J., Đan, M., Radenković, S. (unpubl.) Taxonomy and systematics of three species of the genus Eumerus Meigen, 1822 (Diptera: Syrphidae) new in southeastern Europe. Grković A., Vujić, A., Radenković, S., Chroni, A., Petanidou, T. 2015. Diversity of the genus Eumerus Meigen (Diptera, Syrphidae) on the eastern Mediterranean islands with description of three new species. Annales de la Société Entomologique de France (N.S.), 51(4), 361–373, http://dx.doi.org/10.1080/00379271.2016.1144483
(d)Biodiversity Conservation: What is the species status in Eumerus, e.g. rate of endemism? which are its implications on conservation priorities in the Mediterranean Basin? ACKNOWLEDGEMENTS This study was funded by the grant No. 173002, Ministry of Education, Science and Technological Development of Republic of Serbia. Part of the financial support was provided by the Transnational Access to Research Infrastructures activity in the 7th Framework Programme of the EC under the ExpeER project.
