Divergent leaf traits of medicinally important Cocculus hirsutus (L.) Diels- morphoanatomical and molecular authentication

Cocculus hirsutus (L.) Diels (Menispermaceae) is a perennial liana found mainly in tropical and subtropical regions of the Western Ghats, India. This study investigates the variation in leaf morphology within this species. Five samples were selected based on differences in leaf structure, and morphological analysis of foliar traits (Petiole length, Leaf length and Leaf width) revealed significant variation (p < 0.0001) through one-way ANOVA (F-values for PL = 1247.46, LL = 1303.51, and LW = 1413.46). Principal component analysis (PCA) was used to further explore this diversity. Morphoanatomical features, including leaf epidermis, were examined for accurate species identification. DNA barcoding using matK, rbcL, and combined sequences showed superior results with rbcL and the combined sequence. Maximum likelihood (ML) and Bayesian Inference (BI) were applied to assess species relationships, and the analysis suggested all samples belong to the same species. Nevertheless, discrepancies in the bootstrap and posterior probability values revealed by the rbcL and combined sequences may be attributed to microbial influence or mimicry from the host plant.