Party coordinator O Panneerselvam and co-coordinator K Palaniswami announced "removing" 132 persons from the party's district units of Tirupur, Pudukottai and Dharmapuri.
A total of 132 functionaries were sacked from the party. Tirupur topped the list with 65 being expelled.
The two leaders said in a statement that 49 persons from Pudukottai and 18 from Dharmapuri units were also being removed as they went against the party's principles and brought "disrepute" to the AIADMK.
"Party workers are requested not to have any truck with them," Mr Palaniswami, also the chief minister, and his deputy Mr Panneerselvam, said in a party statement.
Mr Panneerselvam and Mr Palaniswami "relieved" one person from his party post in Dharmapuri.
Stung by the bypoll defeat, the ruling AIADMK on December 25 cracked the whip against nine party functionaries, sacking four of Dhinakaran's aides and expelling five others.
The two leaders had warned of action against those who "betrayed" the party, and had expelled 44 persons yesterday.
In a jolt to the ruling camp, Mr Dhinakaran secured an emphatic win in the December 21 R K Nagar bypoll defeating its candidate E Madhusudhanan by a thumping margin of over 40,000 votes.
The bypoll, necessitated due to the death of sitting MLA, former chief minister J Jayalalithaa, was seen as a litmus test for the 20-month-old AIADMK government.
Mr Dhinakaran, sidelined by the ruling camp a few months ago, contested as an independent candidate.
He had earlier lost his battle for the party symbol 'Two Leaves' to the Palaniswami-led ruling camp and contested the election on pressure-cooker symbol.