"I'm just not ready to do that at this point. I'm not there right now," Ryan told CNN in an interview.
Ryan's stunning statement had its immediate ripple effect inside the party and might have lasting impact on Trump's presidential ambitions, if the division within the Republican establishment are not sorted out soon.
Ryan said the onus for uniting the party now lies on Trump, who after his Indiana primary win and withdrawal from the race by the two opponents -- Ted Cruz and John Kasich -- is the presumptive presidential nominee of the Republican party.
"This is the party of Lincoln, of Reagan, of Jack Kemp. And we don't always nominate a Lincoln and a Reagan every four years, but we hope that our nominee aspires to be Lincoln- and Reagan-esque," the House Speaker said.
He hoped that the candidate "advances the principles of our party and appeals to a wide, vast majority of Americans".
"I am not ready to support Speaker Ryan's agenda. Perhaps in the future we can work together and come to an agreement about what is best for the American people. They have been treated so badly for so long that it is about time for politicians to put them first!" Trump said.
The tensions between Trump and Ryan go beyond temperament, The Washington Post reported.
"They have philosophical differences about the size and scope of government. Ryan champions free-trade agreements, international military engagement, and sweeping overhauls of Social Security and Medicare, whereas Trump is an avowed opponent of recent trade deals, foreign interventions and proposed changes to entitlement programs," the Post reported.
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