To date, the influence of lignin variation on starch accumulation, composition and processing properties in storage roots remains underexplored. We used physiological and biochemical assay to estimate the influence of lignin on starch formation and pasting profiles in 21 cassava storage root species, along with a lignin detection assay using Raman spectroscopy. The lignin content ranged from 3.9 to 7.5% across the species, and the monolignol types primarily consisted of syringyl (S) and guaiacyl (G) units with S/G ratios of 0.16–0.96. The starch content varied greatly among the varieties (26.5–86.9%), with the amylose contents of 12.6–23.9%. Lignin content was negatively correlated with starch content and its composition (