Mugabe told thousands of his supporters at Bindura's Chipadze Stadium, Mashonaland Central that anointing Grace as his successor would be a violation of the Zanu PF constitution.
However, the declaration contradicted his wife who had spoken a few minutes earlier emphasising that the 93-year-old ruler had a right to hand-pick a successor.
Last month the 52-year-old first lady went public demanding that Mugabe anoint a successor as Zanu PF infighting reached frightening levels.
"I will never do that, never," he said. "The party leader will be chosen by the people at congress and as a lawyer, I will respect that."
Mugabe's speech, which appeared like an attempt to cool off tempers after his wife humiliated Mnangagwa, was about the history of factionalism in Zanu PF but also a serious attack on the VP's standing as a politician.
He said he did know how Higher and Tertiary Education minister Jonathan Moyo and Mnangagwa became enemies after plotting against him during the so-called Tsholotsho declaration.
"Prof Moyo chose to support Mnangagwa to take over," Mugabe told the rally.
He acknowledged the existence of G40, which he said was a creation of the likes of national commissar Saviour Kasukuwere and Team Lacoste, but said Mnangagwa always denied that he was a leader of a faction.
"I asked where Lacoste came from and Mnangagwa said he does not know, if it is the cup [I am the boss cup] throw it away. You cannot suffer because of a cup," Mugabe said.
On G40, Mugabe said: "It was started by Kasukuwere after the election of Barack Obama as the American president. Obama as in his 40s and Kasukuwere said we also want a leader in the 40s and called themselves G40.
"But now Obama is gone, that should end. If he called himself G40, he remained in the country and we have a history and processes to follow."
He said Kasukuwere and Masvingo political kingmaker, Josiah Hungwe once clashed in the politburo, with the Psychomotor minister accusing the Local Government minister of using his position as commissar to build his power base to take over power.
Mugabe said Masvingo had serious tribal problems while Midlands had minor issues which could be sorted out.
The Masvingo problems, he said, started during the days of the late Edson Zvobgo, who he said was a self-proclaimed tribalist.
"I met the Masvingo provincial leadership after [Tourism minister Walter] Mzembi was sent by the provincial MPs who called for the axing of [late Provincial Affairs minister] Shuvai Mahofa and Josaya Hungwe," Mugabe said.
He said they tried to resolve the problems but did not manage and a lot of work still needed to be done.
Mugabe also spoke about Moyo's dossier detailing Mnangagwa's alleged plot to seize power.
"Moyo once said in the politburo that he will never support Mnangagwa again," Mugabe said after chronicling Mnangagwa and Defence minister Sydney Sekeramayi's history.
He warned Mnangagwa against being pushed by people into trying to oust him.
Mugabe said Moyo's dossier included allegations that the VP forced former broadcaster Godfrey Majonga to jump from the second floor of a Harare apartment after they clashed over a girlfriend.
Majonga suffered serious injuries and is now wheelchair-bound. Mugabe said Mnangagwa had prepared his own 85-page dossier against Moyo but requested for more time before he presents it to the Zanu PF politburo because he was not well.
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