ssp. A Hybrid Species Between Diploid and .

is considered as most probable wild ancestor of cultivated buckwheat. However, the evolutionary route from to remains to be deciphered. We hypothesized that a hybrid species exists in natural habitats between diploid and . The aim of this research was to determine the phylogenetic position of ssp. and to provide new thoughts on buckwheat evolution. Different methodologies including evaluation of morphological traits, determination of secondary metabolites, fluorescence hybridization (FISH), comparative chloroplast genomics, and molecular markers were deployed to determine the phylogenetic relationship of ssp. with and . The ambiguity observed in morphological pattern of genetic variation in three species revealed that ssp. ancestrale is closely related to and . Flavonoid analysis revealed that ssp. is closely related to . Comparative chloroplast genome analysis further supported the close proximity of ssp. with . Additionally, molecular marker analysis revealed that ssp. exhibits co-dominance with the bands amplified by and . These finding provided supporting evidence in favor of the hypothesis that ssp. is a hybrid species between to , which was probably originated by spontaneous hybridization under natural conditions.