This study aimed to investigate the response of four ecotypes of milk thistle from Isfahan, Kohgiluyeh va Boyer Ahmad, Khuzestan-Omidiyeh, and Hungary to three irrigation regimes (irrigation after 40%, 60%, and 80% maximum allowable depletion of available soil water (evaluated in 2020-2021 under five different planting dates (September 27th, October 17th, November 6th, November 26th, and March 10th) at the research farm of Isfahan University of Technology in Iran. The experiment was conducted as a combined analysis of a split-plot based on randomized complete block design with three replications. The results showed that the highest and the lowest grain yield and shoot dry weight of milk thistle plants were obtained under the October 17th and March 10th planting dates, respectively. Water deficit at both levels decreased grain yield, shoot dry weight, number of flowers per plant, plant height, flowering degree-days, and physiological maturity degree-days, but increased grain and shoot water use efficiencies. The "Kohgiluyeh and Boyer Ahmad" ecotype exhibited the highest seed yield, shoot dry weight, seed/shoot water use efficiency, and harvest index, making it a promising candidate for breeding programs aimed at creating adaptive cultivars of the milk thistle plant. Based on the study's findings, it can be concluded that milk thistle should be planted in the fall season from late September to mid-November under similar climate conditions.