Senator Anning has made a spectacular entrance into federal politics, with the minor party announcing his defection minutes after he was sworn in on Monday.
The Queenslander was third on One Nation's Upper House ticket at the last election and secured 19 first preference votes.
But he was called up after the High Court flicked Malcolm Roberts for being a British dual citizen.
The now-independent said Pauline Hanson wanted him to stand aside to allow Mr Roberts' return.
"She was annoyed that I wasn't going to stand aside for Roberts," he told the ABC.
"After 22 years of solid loyal support of Pauline and her policies, running for her in 1998, being at every campaign that she ever had, I was insulted."
Senator Anning walked out of a party meeting on Monday morning, saying he was subjected to a "vitriolic" verbal attack.
"They demanded that I remove my staff, who had been quite loyal staff with Senator Roberts," he said.
"That was never going to be acceptable to me, and I certainly wasn't going to be working with someone like James Ashby.
"I think Ashby is gradually pushing out all the old guard, the people who've been there for a long time. I think he has other ambitions; I'm certain he's not there for Pauline's good.
"I was forced out of the party pretty much by Pauline, but more or less I would say she was being influenced by James Ashby."
In response, Mr Ashby said: "I'm comfortable with Mr Anning's comments for the moment.
"The only advice I've offered Mr Anning is to sit down and finally have a thorough one-on-one discussion with Senator Hanson before running any further public commentary."
Mr Ashby previously worked for former speaker Peter Slipper before launching legal action against his ex-boss, which was later dropped.
Anning vows to fight for mining, oppose 'renewable energy nonsense'
Senator Anning was flanked by fellow crossbenchers Cory Bernardi and David Leyonhjelm when he entered the chamber to be sworn in.
The new senator said despite the falling out, he would continue to support One Nation's policies and would advocate for regional Queensland.
"I'll be fighting hard for the people of Queensland, the mining, the agriculture, we need water up there [in north Queensland], we need infrastructure projects," he said.
"We need all the things that we had when [former Queensland premier Joh Bjelke-Petersen] was around.
"We had a budget surplus, we had lots of infrastructure going."
Like the man he replaced, Senator Anning has no time for clean energy.
"A lot of our industries can't afford the power with this renewable energy nonsense.
"I don't care how many trillion we spend, we're not going to lower the sea level by a millimetre.
"We've got to start getting a good coal-fired power station in central Queensland, opening up the mines; everyone's getting the use of our coal and we're doing nothing with it."
Senator Hanson, Brian Burston and Peter Georgiou are One Nation's remaining federal representatives.