Cryptic diversity, phenotypic congruence, and evolutionary history of the Leptobotia citrauratea complex (Pisces: Botiidae) within subtropical eastern China

Elucidating the emergence and maintenance of cryptic diversity is a major focus of evolutionary biology. Integrative taxonomy is widely considered as the best practice for delimiting cryptic species and exploring cryptic speciation. This approach is used here to study the Leptobotia citrauratea complex, a group of small-sized loaches so far found in subtropical floodplains and hills of eastern China. A total 170 specimens were collected from 24 sampling sites, encompassing geographical variations and divergent habitas. Six putative species, out of which two are cryptic, were delineated by integrating molecular (two mtDNA and three nuDNA genes) and morphological analyses. These species constituted three ecotypes, exhibiting phenotypic disparities concordant with a habitat transition from high- to low-flow environments. Phenotypic similarities among them were shown to not align with their phylogenetic relationships but closely correlate with habitat utilization. Convergent evolution, driven by similar selective pressure associated with habitat-specific use, likely accounts for the cryptic diversity unveiled in the recently diverging species complex. The diversification of this species complex began in the late Pliocene, coinciding with tectonic activities in the subtropical region of eastern China. Subsequent rapid differentiation during the Pleistocene was possibly driven by regional climate fluctuations. This evolutionary trajectory highlights the crucial roles of geological, climate and ecological factors in shaping biodiversity in this region.