Treasurer Josh Frydenberg has rejected suggestions Scott Morrison was echoing Donald Trump in warning against "negative globalism".
In a major speech to the Lowy Institute overnight, the prime minister argued Australia's international engagement would be driven by national interest rather than "unaccountable" global institutions.
His remarks have been seen as a thinly-veiled swipe at the United Nations, and closely resemble comments the US president made in New York last week.
"Australia charts its own course," Mr Frydenberg told ABC Radio on Friday.
"The prime minister and indeed prime ministers before him have been very clear that we act in our own national interest. We don't have the same national interests as the United States."
Asked if the prime minister was echoing Mr Trump, the treasurer replied: "We're not, I don't accept that assertion."
"What we do is act on behalf of 25 million Australians," Mr Frydenberg said.
"We do disagree with the Americans on a number of issues, not least of which is the big trade tensions that we've got right now between China and the US."
Deputy opposition leader Richard Marles said the prime minister's comments struck the wrong tone.
"Certainly our most important relationship is with the United States and the alliance with the US remains the cornerstone of our national security but I worry about these sorts of comments," he told the Nine Network.
"There are a whole lot of challenges that face the world today which require global solutions, climate change being one of them.
"I think the sort of almost nationalistic talk that we hear at the moment in the speech last night, but also that we heard from the US president a few weeks ago, I actually think is not the right tone at this moment in time."
However, Home Affairs Minister Peter Dutton said Mr Morrison had been very open about the government's priorities.
"I think by being open and frank about the issues is the best way to deal with it," Mr Dutton said.
"In relation to the prime minister's speech, I think he spelled out in a common sense way something that most Australians would agree with."
Australian Associated Press