PEERJ, sa.8, ss.8603, 2020 (SCI-Expanded)
Stylommatophora is one of the most speciose orders of Gastropoda, including terrestrial
snails and slugs, some of which are economically important as human food, agricultural
pests, vectors of parasites or due to invasiveness. Despite their great diversity and
relevance, the internal phylogeny of Stylommatophora has been debated. To date, only
34 stylommatophoran mitogenomes were sequenced. Here, the complete mitogenome
of an invasive pest slug, Arion vulgaris Moquin-Tandon, 1855 (Stylommatophora:
Arionidae), was sequenced using next generation sequencing, analysed and compared
with other stylommatophorans. The mitogenome of A. vulgaris measures 14,547 bp
and contains 13 protein-coding, two rRNA, 22 tRNA genes, and one control region,
with an A + T content of 70.20%. All protein coding genes (PCGs) are initiated with
ATN codons except for COX1, ND5 and ATP8 and all are ended with TAR or Tstop codons. All tRNAs were folded into a clover-leaf secondary structure except for
trnC and trnS1 (AGN). Phylogenetic analyses confirmed the position of A. vulgaris
within the superfamily Arionoidea, recovered a sister group relationship between
Arionoidea and Orthalicoidea, and supported monophyly of all currently recognized
superfamilies within Stylommatophora except for the superfamily Helicoidea. Initial
diversification time of the Stylommatophora was estimated as 138.55 million years
ago corresponding to Early Cretaceous. The divergence time of A. vulgaris and Arion
rufus (Linnaeus, 1758) was estimated as 15.24 million years ago corresponding to one
of Earth’s most recent, global warming events, the Mid-Miocene Climatic Optimum.
Furthermore, selection analyses were performed to investigate the role of different
selective forces shaping stylommatophoran mitogenomes. Although purifying selection
is the predominant selective force shaping stylommatophoran mitogenomes, six
genes (ATP8, COX1, COX3, ND3, ND4 and ND6) detected by the branch-specific
aBSREL approach and three genes (ATP8, CYTB and ND4L) detected by codonbased BEB, FUBAR and MEME approaches were exposed to diversifying selection.
The positively selected substitutions at the mitochondrial PCGs of stylommatophoran species seems to be adaptive to environmental conditions and affecting mitochondrial
ATP production or protection from reactive oxygen species effects. Comparative
analysis of stylommatophoran mitogenome rearrangements using MLGO revealed
conservatism in Stylommatophora; exceptions refer to potential apomorphies for
several clades including rearranged orders of trnW-trnY and of trnE-trnQ-rrnS-trnMtrnL2-ATP8-trnN-ATP6-trnR clusters for the genus Arion. Generally, tRNA genes tend
to be rearranged and tandem duplication random loss, transitions and inversions are
the most basic mechanisms shaping stylommatophoran mitogenomes.