The president made the threat while a bipartisan group of US lawmakers were visiting Denmark in the hope of easing tensions over the territory.
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Trump said at a White House meeting that he "may put a tariff on countries if they don't go along with Greenland", which is a self-governing territory controlled by Denmark.
He did not say which countries might be hit with new tariffs, or what authority he would invoke to use such import taxes in pursuit of his goal.
Along with Denmark and Greenland, other countries oppose his plans, and many in the US have expressed scepticism about an acquisition. As Trump spoke, a bipartisan congressional delegation was visiting Greenland to show support for the territory.
The 11-member group included Republicans who voiced concerns about the president's calls for the US to somehow acquire Greenland for national security reasons. They met MPs as well as Danish Prime Minister Mette Frederiksen and her Greenlandic counterpart Jens-Frederik Nielsen.
Group leader Senator Chris Coons, a Democrat, said their trip was to listen to the locals and take their views back to Washington "to lower the temperature".
Trump has said Greenland is vital for US security - and Washington would get it "the easy way" or "the hard way" - an apparent reference to buying the island or taking it by force.
"I may put a tariff on countries if they don't go along with Greenland, because we need Greenland for national security," Trump said at Friday's White House meeting on rural healthcare.
Greenland is sparsely populated but resource-rich and its location between North America and the Arctic makes it well placed for early warning systems in the event of missile attacks and for monitoring vessels in the region.
The US already has more than 100 military personnel permanently stationed at its Pituffik base - a missile-monitoring station on Greenland's north-western tip that has been operated by the US since World War Two.
Under existing agreements w