Suberin polyaliphatics (SPA) accumulation on the closing layer plays a significant role in wound healing of fruit and vegetables. However, no literature is available on abscisic acid (ABA) effect on yam tuber healing in relation to SPA biosynthesis. In this study, ABA treatment decreased weight loss and disease incidence of yam tubers, maintained the integrity of cell membrane, and promoted the accumulation and polymerization of SPA monomers such as fatty acids (FAs), ω-hydroxyacid (ω-OHA) and α, ω-dicarboxylic acid (α, ω-diA). Gene expression of β-ketoacyl CoA synthases (DoKCS2, DoKCS6), cytochrome P450 fatty acid ω-hydroxylases (DoCYP86A1, DoCYP94B3), Glycerol-3-phosphate acyltransferases (DoDPAT5, DoGPAT7) and feruloyl-CoA transferase (DoFHT), which were associated with the biosynthesis of SPA monomers, were enhanced by ABA treatment. ABA improved gene expression of ATP-binding cassette transporters (DoABCG2, DoABCG6) and GDSL-type esterase/lipases (DoGELP38, DoGELP51, DoGELP96), which might facilitate the transport and polymerization of SPA monomers. Contrarily, applied fluridone (FLD, an inhibitor of ABA biosynthesis) exhibited an opposite effect to ABA treatment. The results indicate that ABA accelerates yam tuber healing via positively regulating SPA biosynthesis.