Former US president withdrew in 2018 from the accord, known as the JCPOA, though his defeat to President Joe Biden has raised the prospect of a US return to the deal.
The development was made after he posted a depiction of a man resembling the former US president playing golf in the shadow of a warplane or large drone.
The tweet from Ayatollah Khamenei's account refers to General Qasem Soleimani, who was assassinated by a US drone in Baghdad a year ago.
The Twitter account of Iran’s Supreme Leader on Friday carried the image of a golfer resembling former President Donald Trump apparently being targeted by a drone, vowing revenge over the killing of a top Iranian general in a U.S. drone attack.
The post carried the text of remarks by Ayatollah Ali Khamenei in December, in which he said “Revenge is certain”.
Iranian President Hassan Rouhani urged the incoming U.S. administration on Wednesday to return to a 2015 nuclear agreement and lift sanctions on Tehran, while welcoming the end of “tyrant” President Donald Trump’s era.
U.S. President-elect Joe Biden, who takes office on Wednesday, has said the United States will rejoin the pact, which includes restrictions on Iran’s nuclear work, if Tehran resumes strict compliance.
On Tuesday, alleged air strikes carried out by the United States in Southwest Iraq killed at least nine people and injured dozens.
As reported, witnesses saw American drones flying over al-Qaim, near the Syrian border, despite American security officials denying any strike in the region.
French foreign minister said Iran was building up its nuclear weapons' capacity and it was urgent that Tehran and Washington return to a 2015 nuclear agreement.
Israel could be plotting to derail one of the incoming Biden Administration's signature foreign policy promises.
Officials from the Israeli government are contemplating the efficacy of the same strategy under the incoming Biden Administration, both in terms of public dialogue and behind-the-scenes diplomatic arbitrage.
"At this critical juncture, Iran's action also risks undermining efforts aimed at building upon the existing diplomatic process. We urge Iran to refrain from further escalation and reverse this course of action without delay."