Joice Mujuru came under attack after Grace Mugabe claimed the vice president was extorting money from companies and fomenting factionalism which is threatening to tear the ruling ZANU-PF party apart.
She has also been accused of plotting to assassinate the president, as factions within the party jostle for power when 90-year-old Mugabe steps down or dies.
Today, it emerged that Mujuru's bid to seek reelection to the ruling party's central committee was foiled when ZANU-PF rejected her election papers ahead of a key party congress next week.
Mujuru and powerful Justice Minister Emmerson Mnangagwa were seen as the leading contenders to replace Mugabe, who has been in power since independence from Britain in 1980.
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But several of her close aides have been suspended ahead of the congress, in what appears to be a purge ahead of the congress.
The battle escalated following Grace Mugabe's surprise nomination to lead the powerful women's wing of ZANU-PF, prompting speculation that she wanted the top job herself.
Mujuru's failure to win a place in the central committee means she all but ceases to be in the party's top leadership even before the congress starts on December 3.
"We are witnessing something strange in Zanu-PF where all of us who were in the struggle are being accused of trying to assassinate the president," said Rugare Gumbo, a party veteran who was suspended as spokesman after being accused of conspiring with Mujuru to topple Mugabe.
Mujuru, 59, went by the name Teurai Ropa (Spill Blood) when she served in Mugabe's guerrilla forces during the war against white minority rule in the former Rhodesia.
After independence in 1980, she served in a series of ministerial posts until being appointed vice president in 2004.