Background: Rehmannia glutinosa is a perennial herb belonging to the family Scrophulariaceae and is used in traditional oriental medicine. This study was conducted to evaluate the appropriateness of plug-seedling cultivation for replanting of R. glutinosa. Methods and Results: The seed rhizomes of Rehmannia glutinosa 'Jihwang 1' were harvested in the middle of March, 2018. Direct seedling cultivation was conducted after air curing for 3 days. The seedlings for transplanting were cultivated for 30 (30D), 45 (45D), and 60 days (60D). The mat formation rates of the 30D, 45D and 60D treatment seedling were 81.3%, 100%, and 100%, respectively. Of the tree treatment groups, flowering occurred only in the 45D and 60D treatment groups, and their flowering rates were 7.3 ± 3.06% and 13.3 ± 1.15%, respectively. The agronomic characteristics and active components of the three treatment groups were evaluated in comparison with the DS1 (directly seeded group, control) and DS2 (directly seeded group, replanting) groups. As a result, the leaf length and leaf width of the 30D seedlings were higher than other treatment groups, but the number of leaves per plant was statistically the same. In addition, root diameter and yield of the 30D seedling were statistically the same as DS1. Moreover the catalpol contents of 30D, 45D, DS1 and DS2 group seedlings were 2.23%, 1.86%, 2.50% and 2.08%, respectively. Conclusions: Taken together, these findings suggest it is feasible to use the plug-seedling cultivation method for replanting of R. glutinosa. seedlings. However, further evaluation of the economic aspect is needed.