One day after warning the GOP that his "shackles" are off, Republican presidential nominee Donald Trump Donald John TrumpOmar fires back at Trump over rally remarks: 'This is my country' Pelosi: Trump hurrying to fill SCOTUS seat so he can repeal ObamaCare Trump mocks Biden appearance, mask use ahead of first debate MORE floated the prospect of a "sinister deal" preventing Speaker Paul Ryan Paul Davis RyanKenosha will be a good bellwether in 2020 At indoor rally, Pence says election runs through Wisconsin Juan Williams: Breaking down the debates MORE (R-Wis.) from coming to his defense.

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Trump at a Florida rally on Wednesday questioned why Ryan, who effectively conceded Trump's White House chances during a call with lawmakers this week, wouldn't come out and congratulate him after Sunday's presidential debate.

"Wouldn't you think Paul Ryan would call and say, 'good going'?" Trump asked in Ocala.

"You'd think they'd say, 'Great going, Don. Let's go, let's beat this crook. Let's beat her. We've got to stop her.' No, he doesn't do that. There's a whole deal going on. We're going to figure it out. I always figure things out. There's a whole sinister deal."

It's the latest evidence of a deepening fracture between the GOP nominee and his party's establishment less than one month before the election. It all comes while frantic Republicans face the fallout of Friday's release of damaging audio in which Trump talks about groping women without their consent.

More than two-dozen lawmakers called on Trump to withdraw as the nominee and leave the top of the ticket to his running mate, Mike Pence Michael (Mike) Richard PenceButtigieg stands in as Pence for Harris's debate practice Trump argues full Supreme Court needed to settle potential election disputes Pence adviser knocks ex-staffer who criticized Trump on COVID-19 MORE. But Trump has emphatically sworn off the idea, telegraphing all-out warfare against the party with a series of Tuesday morning tweets declaring that the "shackles" put on him by establishment Republicans are now off.

The discontent all comes as Democratic nominee Hillary Clinton Hillary Diane Rodham ClintonButtigieg stands in as Pence for Harris's debate practice Senate GOP sees early Supreme Court vote as political booster shot Poll: 51 percent of voters want to abolish the electoral college MORE continues to expand her lead at the polls. Those top-of-the-ticket gains have prompted Republicans and Democrats alike to begin to wonder if the fallout could deliver the Senate to the Democrats and even put House control in play.

Trump has shown no sign of playing nice with the party, putting him at odds with the Speaker, who agreed to take the post last year in the hopes of building party unity.

Ryan repudiated Trump's 2005 comments in a statement over the weekend. But while the Wisconsin Republican did not withdraw his endorsement officially, he told lawmakers this week he would no longer defend Trump or campaign with him. Instead, Ryan said that he would focus on maintaining the House majority as a check on Clinton if she wins.

Ryan's decision has drawn the ire of Trump and his allies.

The GOP nominee tore into Ryan on Fox News's "The O'Reilly Factor" on Tuesday night, saying he doesn't want Ryan's support and that the Speaker is the only person who could lose to President Obama in a negotiation.

And while a number of lawmakers criticized Ryan for his decision during this week's private lawmaker call, Rep. Jim Bridenstine James (Jim) Frederick BridenstineNASA publishes Artemis plan to land first woman, next man on moon NASA is in the market for moon rocks Overnight Defense: Trump hosts Israel, UAE, Bahrain for historic signing l Air Force reveals it secretly built and flew new fighter jet l Coronavirus creates delay in Pentagon research for alternative to 'forever chemicals' MORE (R-Okla.) tweeted Wednesday that he could no longer support Ryan if the Speaker doesn't back Trump.

Trump's inner circle includes at least one operative who has long advocated for Ryan's political destruction, The Hill reported exclusively on Tuesday.

Trump campaign CEO Steve Bannon, then head of conservative news website Breitbart, told a reporter that he hoped to see Ryan "gone" by this past spring.

--This report was updated at 2:56 p.m.