Darwin was fascinated by the transportation of land snails across great swathes of open ocean by birds--he even immersed snails in sea water to see how long they would survive. Here we follow a molecular phylogenetic trail that reveals the incredible transequatorial dispersal of the land snail Balea from Europe to the Azores and the Tristan da Cunha islands, and back again. This long-distance dispersal is unexpected for what are proverbially considered the most pedestrian of creatures.
We sequenced 665bp of the Cytochrome C Oxidase I (COI) barcoding marker for 257 specimens and 482bp of Elongation Factor 1-α (EF1-α) for 237 specimens belonging to the leafmining subgenus Ectoedemia (Ectoedemia) in the basal Lepidopteran family Nepticulidae. The dataset includes 45 out of 48 West Palearctic Ectoedemia s. str. species and several species from Africa, North America and Asia. Both COI and EF1-α proved reliable as an alternative to conventional species identification for the majority of species and the combination of both markers can aid in species validation. A clear barcode gap is not present, and in some species large K2P intraspecific pairwise differences are found, up to 6.85% in COI and 2.9% in EF1-α. In the Ectoedemia rubivora species complex, the species E. rubivora, E. arcuatella and E. atricollis share COI barcodes and could only be distinguished by EF1-α. Diagnostic base positions, usually third codon positions, are in this and other cases a useful addition to species delimitation, in addition to distance methods. Ectoedemia albifasciella COI barcodes fall into two distinct clusters not related to other characters, whereas these clusters are absent in EF1-α, possibly caused by mtDNA anomalies or hybridisation. In the Ectoedemia subbimaculella complex, both sequences fail to unequivocally distinguish the species E. heringi, E. liechtensteini, E. phyllotomella and one population of E. subbimaculella. DNA barcodes confirm that North American Ectoedemia argyropeza are derived from a European introduction. We strongly advocate the use of a nuclear marker in addition to the universal COI barcode marker for better identifying species, including cryptic ones.
Phylogenetic relationships in species complexes and lineages derived from rapid diversifications are often challenging to resolve using morphology or standard DNA barcoding markers. The hyper-diverse genus Lepanthes from Neotropical cloud forest includes over 1200 species and many recent, explosive diversifications that have resulted in poorly supported nodes and morphological convergence across clades. Here, we assess the performance of 446 nuclear-plastid-mitochondrial markers derived from an anchored hybrid enrichment approach (AHE) coupled with coalescence- and species network-based inferences to resolve phylogenetic relationships and improve species recognition in the Lepanthes horrida species group. In addition to using orchid-specific probes to increase enrichment efficiency, we improved gene tree resolution by extending standard angiosperm targets into adjacent exons. We found high topological discordance among individual gene trees, suggesting that hybridization/polyploidy may have promoted speciation in the lineage via formation of new hybrid taxa. In addition, we identified ten loci with the highest phylogenetic informativeness values from these genomes. Most previous phylogenetic sampling in the Pleurothallidinae relies on two regions (ITS and matK), therefore, the evaluation of other markers such as those shown here may be useful in future phylogenetic studies in the orchid family. Coalescent-based species tree estimation methods resolved the phylogenetic relationships of the L. horrida species group. The resolution of the phylogenetic estimations was improved with the inclusion of extended anchor targets. This approach produced longer loci with higher discriminative power. These analyses also disclosed two undescribed species, L. amicitiae and L. genetoapophantica, formally described here, which are also supported by morphology. Our study demonstrates the utility of combined genomic evidence to disentangle phylogenetic relationships at very shallow levels of the tree of life, and in clades showing convergent trait evolution. With a fully resolved phylogeny, is it possible to disentangle traits evolving in parallel or convergently across these orchid lineages such as flower color and size from diagnostic traits such as the shape and orientation of the lobes of the petals and lip.
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