Aney, at an event in Jalna yesterday where he disbursed relief to families of farmers who committed suicide, said Marathwada bore more injustice than Vidarbha and should therefore be independent.
He also called people of Marathwada to start a movement for formation of a separate state.
In December last year, Aney had called for referendum on the issue of carving out separate Vidarbha state.
The signatories to it include former Chief Minister Prithviraj Chavan of Congress, Dilip Walse Patil of NCP and SP leader Abu Azmi.
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"By making such irresponsible and provocative statement, Aney has violated provisions of Article 165 of the Constitution," Walse Patil said.
"People of Maharashtra should know who is behind Aney's remarks. Are those who appointed him behind his tirade," he said, in a veiled reference to Aney's close relations with Chief Minister Devendra Fadnavis.
His remark sparked uproar in the Assembly ,which was adjourned thrice and later for the day as Congress-NCP and Shiv Sena raised slogans and demanded the AG's resignation.
In the Upper House, amidst the uproar, an Opposition member sought to know if the government will invoke 'sedition' charge against the Advocate General.
State Revenue Minister Eknath Khadse said in the Legislative Council that the government is ready for a debate over the issue and a decision on Aney's statement would be taken within an hour's time.
Panel Chairman Muzaffar Hussain, however, chose to adjourn the House for the day.
Leader of Opposition in the Council, Dhananjay Munde, who had initially moved the adjournment motion, called for an immediate clarification by Chief Minister Devendra Fadnavis over the issue and demanded Aney's ouster.
"After batting for a separate statehood for Vidarbha, he now wants separate statehood for Marathwada. Tomorrow he will demand a separate Khandesh and then want Mumbai to be separated from the state," Munde said.