An Empirical Demonstration of Unsupervised Machine Learning in Species Delimitation

Abstract One major challenge to delimiting species with genetic data is successfully differentiating species divergences from population structure, with some current methods biased towards overestimating species numbers. Many fields of science are now utilizing machine learning (ML) approaches, and in systematics and evolutionary biology, supervised ML algorithms have recently been incorporated to infer species boundaries. However, these methods require the creation of training data with associated labels. Unsupervised ML, on the other hand, uses the inherent structure in data and hence does not require any user-specified training labels, thus providing a more objective approach to species delimitation. In the context of integrative taxonomy, we demonstrate the utility of three unsupervised ML approaches, specifically random forests, variational autoencoders, and t-distributed stochastic neighbor embedding, for species delimitation utilizing a short-range endemic harvestman taxon (Laniatores, Metanonychus ). First, we combine mitochondrial data with examination of male genitalic morphology to identify a priori species hypotheses. Then we use single nucleotide polymorphism data derived from sequence capture of ultraconserved elements (UCEs) to test the efficacy of unsupervised ML algorithms in successfully identifying a priori species, comparing results to commonly used genetic approaches. Finally, we use two validation methods to assess a priori species hypotheses using UCE data. We find that unsupervised ML approaches successfully cluster samples according to species level divergences and not to high levels of population structure, while standard model-based validation methods over-split species, in some instances suggesting that all sampled individuals are distinct species. Moreover, unsupervised ML approaches offer the benefits of better data visualization in two-dimensional space and the ability to accommodate various data types. We argue that ML methods may be better suited for species delimitation relative to currently used model-based validation methods, and that species delimitation in a truly integrative framework provides more robust final species hypotheses relative to separating delimitation into distinct “discovery” and “validation” phases. Unsupervised ML is a powerful analytical approach that can be incorporated into many aspects of systematic biology, including species delimitation. Based on results of our empirical dataset, we make several taxonomic changes including description of a new species.