Leaves of American mayapple (Podophyllum peltatum) are being investigated as an alternative and renewable source of podophyllotoxin, a pharmaceutical compound used in the manufacture of several drugs. This study examined long-term performance of mayapple populations subjected to different harvest strategies. A naturally occurring population in shade was subjected to leaf removal treatments of frequency (every year, every 2nd or 3rd year) or timing (early or late season). Plots were 1.0 m2, established during Spring 2001, and treatments were applied from 2001 to 2004. Control plots not previously harvested were also included each year. Plants did not tolerate the severest of leaf removal treatments: early harvest time in combination with annual harvest frequency. Early annual harvests reduced total leaf dry mass and total leaf area in a quadratic manner. Late harvest conducted annually, and early harvest conducted every other year, also reduced leaf dry mass and area but not as much as early annual harvest. Plants harvested every year, early, or early every year produced fewer sexual shoots than other treatment combinations. Contents of α-peltatin, β-peltatin, and total lignans were higher for leaves harvested early than those harvested late during each year of the study, demonstrating that lignan contents were affected by leaf age and not treatment. In conclusion, our results for plants grown in shade show that leaves can be removed late in the growing season every 2nd or 3rd year or early every 3rd year without reducing long-term performance of the population. This is more restrictive than that reported for populations in full sun where plants tolerated late harvests every year.