Ryan Braun's 65-game suspension last month and previous punishments bring to 18 the total number of players disciplined for their relationship to Biogenesis of America, a closed anti-aging clinic in Florida accused of distributing banned performing-enhancing drugs.
The harshest penalty was reserved for Rodriguez, a three-time Most Valuable Player and baseball's highest-paid star. His suspension covers 211 games, starting Thursday, and he is expected to appeal.
The New York Yankees slugger admitted four years ago that he used performance-enhancing drugs while with Texas from 2001-03 but has repeatedly denied using them since.
Rodriguez was suspended under both the drug agreement and labor contract.
MLB said the drug penalty was for "his use and possession of numerous forms of prohibited performance-enhancing substances, including testosterone and human growth hormone over the course of multiple years."
His penalty under the labor contract was "for attempting to cover up his violations of the program by engaging in a course of conduct intended to obstruct and frustrate the office of the commissioner's investigation."
Rodriguez has until Thursday to appeal, and if he does so, he will remain eligible to play until a decision by the arbitrator.
"Those players who have violated the program have created scrutiny for the vast majority of our players, who play the game the right way," baseball Commissioner Bud Selig said. "We continue to attack this issue on every front - from science and research, to education and awareness, to fact-finding and investigative skills."