The role of western Mediterranean islands in the evolutionary diversification of the Spotted Flycatcher (Muscicapa striata) Pons J.-M.(1,2,*) , Thibault J.-C(1) , Aymí R.(3) , Grussu M(4)., Muntaner J.(5) , Olioso G.(6) , Sunyer J.R.(7) , Touirhi M.(8) , Fuchs J.(1,2*) (1) UMR 7205 Institut de Systématique, Evolution, Biodiversité, CNRS, MNHN, UPMC, EPHE, Sorbonne Universités, Département Systématique et Evolution, Muséum National d’Histoire Naturelle, CP 51, 55 rue Buffon, 75231 Paris Cedex 05, France. (2) UMS MNHN/CNRS 2700 Outils et Méthodes de la Systématique Intégrative (OMSI), Muséum National d’Histoire Naturelle, 57 rue Cuvier, 75231 Paris Cedex 05, France. (3) Institut Català d'Ornitologia, Museu de Zoologia, Passeig Picasso, s/n, 08003 Barcelona, Spain. (4) Gruppo Ornithologico Sardo, C.P. 160/C 09045, Quartu Sant’ Elena, Cagliari, Italy. (5) Servei d'Agents de Medi Ambient, Unitat de Flora i Fauna, Direcció Gral. De Medi Natural, Educació Ambientalbi Canvi Climàtic, Conselleria d'Agricultura, Medi Ambient i Territori del Govern de les Illes Balears, Spain. (6) 248 rue de l’industrie, 11210 Port-La-Nouvelle, France. (7) Servei d’Agents de Medi Ambient, Conselleria d’Agricultura, Medi Ambient i Territori. Govern de les Illes Balears, Gremi Corredors 10. Polígon Son Rossinyol. 07009 Palma, Spain. (8) Unité de recherche de Bioécologie Animale et Systématique Evolutive, Département de Biologie, Faculté des sciences de Tunis, Université de Tunis EL Manar, Tunisia. (*)
[email protected],
[email protected]
INTRODUCTION
The Spotted Flycatcher is a Palaearctic long distance migratory passerine with a widespread range covering North Africa and a large part of the Eurasian continent (from Western Europe up to Mongolia eastward; Fig, 1) during the breeding season and most of sub-Saharan Africa during winter. Four mainland subspecies including the nominate in Europe as well as two insular subspecies (M. s. tyrrhenica in Corsica and Sardinia and M. s. balearica in the Balearics) are currently recognized based on morphology. In this study, we investigated the genetic relationships among insular and mainland populations of the Spotted Flycatcher using a multilocus approach.
OBJECTIVES OF THE STUDY In this study, we addressed the following questions: What role did the western Mediterranean islands play in the evolutionary history of the Spotted Flycatcher? Did Pleistocene climatic oscillations influence the extant genetic structure of the Spotted Flycatcher? Does the current taxonomy, based on morphology, reflect the phylogenetic relationships and genetic divergence among subspecies?
MATERIAL AND METHODS
We used multilocus data and Bayesian analyses to infer the phylogenetic relationships among the Spotted Flycatcher subspecies and to estimate the divergence time among the primary lineages. Individual locus and multilocus allele networks were reconstructed to visualize genetic relationships among insular and mainland populations. Population genetic estimates were obtained using the mitochondrial COI for 66 spotted flycatchers.
Muscicapa striata balearica (photo: Josep R. Sunyer)
RESULTS MtDNDA Two strongly supported clades, that likely diverged from each other during the the mid-Pleistocene (COI genetic distance 3.5%), within M. striata (Fig. 2): • 1
2
2 4
1
• The second clade included all individuals sampled on the mainland (western Europe to eastern Russia and extending to North Africa). This group was further divided into two sub-groups (Northern Africa/Spain and Eurasia) that diverged from each other by 1.3% (CO1 sequences). No obvious genetic structure was observed within the Eurasian clade despite its broad distribution,
1 M. s. neumanni
1
The first clade included individuals that belong to the insular balearica (Balearic Is) and tyrrhenica (Corsica, Sardinia) subspecies (including two mainland birds breeding along the Tyrrhenian coastline, Italy). The COI genetic distance between balearica and tyrrhenica was 1.7 %.
1 1
MNHN JMP193 Corsica
1 M. s. striata
3
1
1 16 1
MNHN JMP297 Corsica 3
2
2
1
1
1
1.0
MNHN JMP658 3A90470 Sardinia
M. s. neumanni 2 4
MNHN JMP747 Italy
RESULTS 1.0 1.0
10
MNHN JMP752 Italy
1.1 Mya (0.9-1.4)
16 M. s. balearica
tyrrhenica
3
MNHN JMP670 EV5808 Balearic Is
Muscicapa striata
M. s. tyrrhenica
balearica
1.0
1.0
MNHN JMP678 MC4963 Balearic Is MNHN JMP754 1Z12537 Spain
1.0
Figure 1: Breeding distribution of the spotted flycatcher (sample size in dots). Blue dots indicate COI sequences retrieved from Genbank and included in the phylogeographic analyses. Wintering range (not shown) covers a large part of sub-Saharan Africa up to the southern tip of the continent.
MNHN JMP323 Tunisia
1.0 MNHN JMP387 Tunisia
1.0 MNHN JMP745 Italy
RESULTS NucDNDA
1.0
Phylogenetic relationships among nuclear alleles were generally poorly resolved due to low level of variation
2.6 Mya (2.1-3.2)
The proportion of shared nuclear alleles between insular and mainland lineages was low (0/9 for FGB, 1/12 for MB, 1/13 for TGFb2, 2/17 for LDH; Fig. 3) Such a low proportion of shared alleles may be explained by lack of recent gene flow between insular and mainland lineages
1.0
ZMUC 147532 Denmark
0.97
UWBM 61090 Russia
striata
MNHN SPOT13430 France
1.0
The multilocus network obtained revealed strong genetic divergence between insular (balearica, tyrrhenica) and the European mainland subspecies (striata)
Muscicapa aquatica Muscicapa adusta
0.95
Muscicapa cassini
M. s. balearica, M. s. tyrrhenica (Mediterranean Is, Tyrrhenian coast)
FGB
Muscicapa muttui
0.1
Figure 2: 50% majority-rule consensus tree obtained from the Bayesian analyses of three concatenated mitochondrial loci (COI, ND2, Cytb) Values next to nodes are Bayesian posterior probabilities.
TGFb2
MAIN CONCLUSIONS
M. s. striata (mainland)
0.1
• Western Mediterranean islands played a major role in the evolution of the Spotted Flycatcher
M. sethsmithi M. adusta
•
M. aquatica
The two primary lineages likely diverged from each other during the Pleistocene (1.1 Mya)
• Genetic divergence between the insular and mainland lineages is of the same order of magnitude as in traditionally recognized Muscicapidae species (Ficedula, Sætre et al. 2001)
Multi-locus network
MB
Muscicapa sethsmithi
Balearic Islands
Italy
• Insular subspecies do have different morphology (plumage, wing shape) and migration pattern patterns (Gargallo 1993, Martínez 2011, Viganò and Corso 2015)
Corsica
Western Europe
• Insular breeders are temporally sympatric with mainland migrants without any apparent admixture
Sardinia
Southern Russia
•
The two insular species may be treated as a one polytypic species (Muscicapa tyrrhenica Schiebel, 1910)
Iberian Peninsula
North Africa
•
Attention should be paid to conservation plans for the insular lineage (Balearic flycatchers have the smallest range and likely possess limited level of genetic variation)
LDH
Figure 3: Haplotype networks reconstructed from the four nuclear loci and multilocus network (center) obtained from the four introns (MYO2, FIB5, LDH, TGFB) and the three mitochondrial loci (COI, ND2, Cyt b).
PERSPECTIVES • Increased sampling in Italy to delimit more precisely the distribution range of the insular and mainland lineages along the Tyrrheanian coast and the possible existence of a contact zone • •
Increased sampling on the mainland clade to assess the geogeraphical limits of the Iberian/North African and Eurasian lineages
Use of Next Generation Sequencing data to investigate which parts of the genome are the most differentiated between the insular and mainland lineages (existence of pre or post-zygotic barriers linked to physiology or migratory behavior?)
REFERENCES
Gargallo M. 1993, Identificació de la subspècie balear de menjamosques (Muscicapa striata balearica), Anuari Ornitológic de les balears 8: 21-30, Martínez JL. 2011. Spotted Flycatcher (Muscicapa striata). In Spring migration in the western Mediterranean and NW Africa: the results of 16 years of the Piccole Isole project. In Gargallo G., Barriocanal C., Castany J., Clarabuch O., Escandell R., López-Iborra G., Rguibi-Idrissi H., RobsonD., and Suárez, M. Monografies del Museu de Ciències Naturals 6. Sætre G-P, Borge T, Lindell J, Moum T, Primmer CR, Sheldon BC, Haavie J, Johnsen A, Ellegren H. 2001. Speciation, introgressive hybridization and nonlinear rate of molecular evolution in flycatchers. Mol. Ecol. 10: 737-749. Viganò M and Corso A. 2015. Morphological differences between two subspecies of Spotted Flycatcher Muscicapa striata (Pallas, 1764) (Passeriformes Muscicapidae). Biodiver. J. 6: 271–284.
ACKNOWLEDGMENTS
We are grateful to S. Birks and R. Faucett (UWBM), A. Cibois (MHNG), P. Ericson and U. Johansson (NRM), J. Fjeldså (ZMUC) for providing tissue samples. We also thank J. Bernal (Catalonia), J. Duaigües (Catalonia), A. Lacci (Sardinia), P. Garcies (Mallorca), S. Nicoll (Mallorca), E. Pasquet (France), J. Piacentini (Corsica), G. Ruzeante (Sardinia), M. Suárez (Mallorca) for providing feathers. We are grateful to Y. Simoncini, A. Borzatti , N. Baccetti, I. Cecere, M. Zenatello (ISPRA), who helped with the Italian samples and to F. Charfi (Faculté des Sciences de Tunis) for her invaluable help in the organization of our field trip in Tunisia. Laboratory help was provided by C. Bonillo, J. Lambourdière, J. Utge and D. Zuccon. This study was supported by the network ‘Bibliothèque du Vivant’ funded by the CNRS, the Muséum National d'Histoire Naturelle, the INRA and the CEA (Genoscope) and the ‘ATM Taxonomie Moléculaire’, Muséum National d’Histoire Naturelle.
