"This may be the last chance to solve the country's problems in peace," warned Mario Cossio, governor of the southern Tarija province, as he arrived for the talks in the city of Cochabamba on Thursday.
Roman Catholic church officials and international envoys, including Jose Miguel Insulza, secretary-general of the Organisation of American States, are to observe the talks.
Ivan Canelas, a government spokesman, said "the government thinks an accord in four or five days of continuous work is possible if there is sincere will for dialogue".
Morales has accused the Catholic church of siding with the governors and the US, which he says is inciting protests against him.
'Massacre' allegation
During the talks, the governors are to press their demands for more autonomy and a greater share of revenues from natural gas exports.
Morales' opponents are also demanding the release of Pando province's governor, who was arrested after officials accused him of ordering "a massacre" of peasants last week.
Critics say the talks could deepen Bolivia's divisions if they lead to the government amending a new constitution drafted by an elected constituent assembly dominated by a majority of Morales supporters.
Supporters of Morales, Bolivia's first indigenous president, fear his reforms, which include the redistribution of land to the poor, could be diluted.
"The people could feel deceived and protest. Grass-roots movements will be watching these talks closely,"said Vladimir Alarcon, an assembly delegate from Morales' Movement Toward Socialism party.