The frontline off-spinner produced a magical delivery in the morning session and clean bowled well-set David Warner in the 22nd over.
Warner was batting at 33 when he was duped by Ashwin's ball pitched outside the leg stump and took a sharp turn to dismantle the left-hander's off-stump.
Left-arm spinner Ravindra Jadeja was the most successful among the Indian bowlers with figures of 3/38.
Stunned by the spin, Warner could only stare at the beauty produced by World's No.1 Test bowler.
Australia lost Warner at 52 in the sixth over of the day.
Here's the beauty produced by Ashwin:
Ashwin has now dismissed Warner eight times in Tests, which is more than any other bowler.
India and Australia are facing each other in Bengaluru for the second Test match. India were restricted to 189 by Australia in their first innings.