Breaking the Hybrid Myth of Paphiopedilum wenshanense: Double Bifurcated Divergence Followed by Adaptive Introgression Formed a Morphological Intermediate.

Species with intermediate phenotypes are usually hypothesised to have originated from hybrid speciation. However, other mechanisms, such as double bifurcated speciation with introgression may create the same phenotypes. Clarifying the underlying process generating the observed intermediate morphology is important for conservation efforts, as it provides insights into the species' evolutionary background and adaptability. Here, we utilised genome-wide single nucleotide polymorphism data to test evolutionary history models for Paphiopedilum wenshanense Z. J. Liu & J. Yong Zhang, an orchid species that is widely considered to be a hybrid with intermediate morphology of other two closely related species, P. concolor Pfitzer and P. bellatulum Pfitzer, distributed in Indochina. Our results rejected the hybrid speciation hypothesis and ascribed the intermediate floral phenotypes of P. wenshanese to introgression from P. concolor after it diverged from P. bellatulum during the last glacial maximum. Excluding the geographic covariate, the current genetic mixing rate of P. wenshanense from P. concolor is stronger in the south part of its range and is associated with precipitation in the early summer, indicating that population divergence in P. wenshanense is being driven by adaptive introgression. These genetic patterns also correspond to the observed floral variation across P. wenshaense populations. The introgression from P. concolor, along with the intermediate and varying floral morphologies, may expand the ecological opportunities for P. wenshanense, providing an explanation for why high genetic diversity has been maintained despite a high level of inbreeding and small census population size. Our study provides a framework for future research to investigate the characteristics and underlying mechanisms of intermediate morphologies in general.