Three homoisoflavonoids, including a new compound, 5,7‐dihydroxy‐3‐(4‐methoxybenzyl)‐8‐methyl chroman‐4‐one (1), together with two known compounds, 5,7‐dihydroxy‐3‐(2‐hydroxy‐4‐methoxybenzyl)‐8‐methylchroman‐4‐one (2) and 5,7‐dihydroxy‐3‐(2‐hydroxy‐4‐methoxybenzyl)‐chroman‐4‐one (3), were isolated from the rhizomes of Polygonatum verticillatum (L.) All. (P. verticillatum). Isolated compounds were characterized on the basis of UV, FT‐IR, ESI‐MS, and 1D‐, 2D‐NMR data. Further, different extract fractions and pure compounds from Polygonatum verticillatum were screened for their antimicrobial potential. Among three pure compounds, compound 2 was found most potent with good zone of microbial growth inhibition as compared to the standards.