NMR methods can be used to delineate detailed structural and regiochemical information on the plant cell wall and elucidate biochemical incorporation pathways. Maize lignin isolated in high yield from rind tissue of stem internodes contained high amounts of esterified p-coumaric acid. Available literature indicated that acylation of the lignin by p-coumaric acid was at the alpha- or gamma-position of the lignin side chain, implicating two diverse biochemical pathways. Application of C-13-H-1 correlative NMR experiments to this maize lignin, synthetic alpha- and gamma-p-coumaroylated lignin model compounds, and a specifically labeled synthetic coniferyl alcohol/coniferyl p-coumarate dehydrogenation polymer (DHP) unambiguously revealed that p-coumaric acid is attached exclusively at the gamma-position. The biochemical implication is that lignin acylation occurs by enzymatically controlled pre-acylation of lignin monomers which are subsequently incorporated into the lignin polymer via oxidative coupling. "Opportunistic" acylation at the alpha-position via quinone methide intermediates is insignificant.