Prime Minister Julia Gillard sided with Israel in opposing Palestinian membership of a United Nation's cultural organisation, despite pleas from her Foreign Minister Kevin Rudd for Australia to abstain on the vote.
Australia was one of 14 countries to vote against Palestine's membership of UNESCO.
The vote was successful after it was backed by 107 members, while Britain and New Zealand were among 52 to abstain.
Mr Rudd wrote to the prime minister urging Australia to abstain from voting, Fairfax reported.
The UNESCO vote is seen as a guide as to where Ms Gillard - a strong supporter of Israel - stands on Palestine's bid for statehood.
Support for Palestine is believed to be important to Australia securing a spot in the Security Council next year, a long-time goal of Mr Rudd's.
The Security Council is expected to decide whether Palestine will become a UN member on November 11 and Washington is threatening to veto any resolution.
A spokeswoman for Ms Gillard said the best way to achieve a Palestinian state was through a negotiated peace.
"We voted against the resolution because it is premature for a subsidiary body of the United Nations (such as UNESCO) to consider this matter while it is still being considered by the United Nations Security Council," she told Fairfax.
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