Excluding spatial sampling bias does not eliminate over-splitting in
DNA-based species delimitation analyses
Abstract
DNA-barcoding and DNA-based species delimitation are major tools in DNA
taxonomy. Sampling has been a central debate in this context, because
the geographical composition of samples affect the accuracy and
performance of DNA-barcoding. Performance of complex DNA-based species
delimitation is to be tested under simpler conditions in absence of
geographic sampling bias. Here, we present an empirical data set sampled
from a single locality in a Southeast-Asian biodiversity hotspot (Laos:
Phou Pan mountain). We investigate the performance of various species
delimitation approaches on a megadiverse assemblage of herbivore chafer
beetles (Coleoptera: Scarabaeidae) to infer whether species delimitation
suffers in the same way from exaggerate infraspecific variation despite
the lack of geographic genetic variation that led to inconsistencies
between entities from DNA-based and morphology-based species inference
in previous studies. For this purpose, a 658 bp fragment of the
mitochondrial cytochrome c oxidase subunit 1 (cox1) was analysed for a
total of 186 individuals of 56 morphospecies. Tree based and distance
based species delimitation methods were used. All approaches showed a
rather limited match ratio (max. 77%) with morphospecies. PTP and TCS
prevailingly over-splitted morphospecies, while 3% clustering and ABGD
also lumped several species into one entity. ABGD revealed the highest
congruence between molecular operational taxonomic units (MOTUs) and
morphospecies. Disagreements between morphospecies and MOTUs were
discussed in the context of historically acquired geographic genetic
differentiation, incomplete lineage sorting, and hybridization. The
study once again highlights how important morphology still is in order
to correctly interpret the results of molecular species delimitation.