Or SNP 629, we located two mismatches (i.e., two xeric individuals bearing a homozygote G, instead of A, or A/G, as observed within the rest of xeric specimens; Table 1). When blasting these two contigs applying the blastn tool Talsaclidine mAChR against lepidopteran sequences40, hits against butterfly mRNAs had been found: for RPR 73401 Autophagy sequence 29, blastn retrieved a mRNA sequence coding for RING finger and SPRY domain-containing protein 1-like from Papilio polytes (XM_013291785.1), as well as an mRNA coding for elongator complex protein 1 from Papilio machaon (XM_014513286.1); for sequence 629, blastn retrieved an anonymous mRNA from Bombyx mori (AK405939.1) too as a putative uncharacterized protein from Papilio xuthus (XM_013315886.1). Wolbachia infection was found in all analyzed samples–32 among 2,116 contigs had been of Wolbachia origin. For all 32 contigs, there was only one particular single allele in all analyzed specimens.DiscussionIn this study, we investigated the genetic coherence of 3 ecotypes from the emblematic butterfly Maculinea alcon, utilizing Next-Generation-Sequencing solutions. Contrary to prior studies, applying classical population genetics approaches to a local-scale sampling encompassing two ecotypes, we employed whole-genome RAD-seq and investigated the ecological genomics of 3 ecotypes at a large geographical scale. Furthermore, we applied BayeScan to search for loci under choice. Similarly to preceding studies20,27,30,34?six, our genome-level SNP analyses didn’t reveal sturdy genetic differentiation amongst the three ecotypes but mainly geographical structuring (see Fig. 1).SCIEnTIFIC REPORTS 7: 13752 DOI:10.1038/s41598-017-12938-www.nature.com/scientificreports/This suggests that ecological shifts within this species will not be linked with lineage sorting and usually do not represent an early stage of speciation, in contrast to processes occurring in several butterflies and other herbivorous arthropods41?3. Our result is in agreement with previous analyses based on spatially a lot more restricted datasets of M. alcon, showing no important genetic structure associated with all the respective ecotypes and larger local genetic relatedness in the two low-altitude ecotypes of M. alcon, which includes syntopically occurring forms, than of populations of each and every ecotype from geographically distant localities30,35,44,45. Similarly, two individuals from our study (a single hygric – 10-B443 and one particular xeric – 10-C370), originating from the similar locality as studied by Tartally et al.30 did not cluster together, also suggesting a mixed origin of this population. Such a pattern is paralleled by observations in other species in the genus Maculinea, as for instance in M. arion, in which the two phenological types differ in morphology and ecology but not genetically46. Initially, it was thought that Maculinea species or forms rely on a single host ant and it was suggested that social parasitism and connected specific life cycle might chiefly impact the evolution, speciation and genetic background of these butterflies32. On the other hand, it has been subsequently shown that the usage of ant hosts may differ among localities, and could possibly be adapted to exploiting a number of species or be switched to other species when the preferred host isn’t present or occurs only in low densities17. Thus, the hypothesis with regards to the influence with the ant host was revisited and some authors suggested that adaptation to the host plant (and corresponding habitat) is much more likely to drive the evolution of Maculinea than would do adaptat.