Barnaby Joyce will recommended Nationals voters in his New England electorate preference One Nation's Richard Thomas second on their ballots.
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ACM understands the preferencing decision was a captain's call by the deputy prime minister.
Meanwhile, Pauline Hanson's One Nation announced this week that it will preference Labor over the Liberals in a number of key seats as a "wake-up call" to the party.
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The Nationals Party finalised its New England how-to-vote card this week.
Eight candidates are contesting the seat of New England against the Nationals leader and incumbent.
The card recommends Nationals voters put One Nation candidate Mr Thomas, United Australia Party candidate Cindy Duncan and Pavlo Samios from the Liberal Democrats before Labor candidate Laura Hughes, who is in sixth slot.
Carol Sparks from the Greens is in seventh slot and independent Natasha Ledger is last.
Mr Joyce told media today that preference agreements "are inherently important" for which party forms government.
He said parties "sit down and work out where is the person who has the closest philosophy to where they are ... and I've tried to do that."
"Same as Labor and the Greens, we rely on One Nation preferences. Certainly the Liberal party relies on us to give them the numbers to be the government. That's just being factual," he said.
One Nation, which is led by Queensland Senator Pauline Hanson, put Labor above the coalition in key seats on its own how-to-vote cards.
Ms Hanson said her party planned to target "left-leaning Liberals" in seats in Tasmania, Melbourne, Sydney and South Australia.
"The Liberals need a wake-up call and I'm more than happy to provide it," she said.
She said the National Party had used "some common sense when it came to preferencing One Nation ahead of other parties" in the New England
"There will be a number of farmers and regional industry groups upset with the National Party and their decision to support Scott Morrison's net-zero policy by 2050, so it was important that One Nation stood a candidate in New England to offer those voters an alternative at this election," she said.
Mr Joyce said he held preference negotiations with the party, but ultimately it was a decision for One Nation.
The Nationals leader first took the seat in the 2013 wipeout after Independent MP Tony Windsor retired. He held it against a challenge by Mr Windsor in 2016.
The Division of New England is one of the safest seats the National Party holds - the ninth safest coalition seat in the country - and has been held by the party for a century, aside from a 12-year period under Mr Windsor.
He regained the party leadership in 2021 after successfully challenging then-leader Michael McCormack.
Parties do not directly affect the flow of preferences in lower house seats, but typically hand out how-to-vote-cards which make a recommendation as to how their supporters should number their ballot. Voters are free to choose whether to follow the card or not.