Yashasvi Jaiswal retired hurt after hitting an attacking century to extend India's lead over England to 322 after the tourists' batting collapse in the third Test on Saturday.
India reached 196-2 in their second innings at stumps on day three in Rajkot with Shubman Gill (65) and nightwatchman Kuldeep Yadav (three) batting at the close of play.
India have been left with 10 players to bat and bowl after Ashwin departed the match late Friday due to a family emergency. Devdutt Padikkal came in as substitute fielder.
Jaiswal, struggling with his back, retired hurt on 104, and the next man Rajat Patidar was out for a duck from 10 balls.
Fast bowler Mohammed Siraj led the bowling charge with figures of 4-84 to help bowl out England for 319 after the visitors collapsed from 299-5.
Skipper Rohit Sharma departed early on 19, but the left-handed Jaiswal stood strong to build the innings along with Gill before the opener changed gears.
Jaiswal smashed veteran pace bowler James Anderson for a six and two fours on successive balls and kept up the charge against opposition spinners with his sweep and reverse sweep.
He raised his third Test ton in just his seventh match with a boundary off fast bowler Mark Wood as he jumped, roared and blew kisses to an applauding crowd.
The 22-year-old, who hit a match-winning 209 in the previous Test, leads the series batting with 435 runs after his 155-run stand with Gill.
Rohit, who hit 131 in the first innings, missed a sweep off part-time spinner Joe Root and England successfully reviewed the decision after the on-field umpire denied the call.
Earlier Ben Duckett hit 153 and skipper Ben Stokes made 41 in his 100th Test but their innings came to a quick halt in the second session.
Siraj wiped out the tail with help from Ravindra Jadeja, who along with fellow spinner Kuldeep took two wickets each.
Left-hander Stokes attempted to hit back after Duckett's departure but fell to Jadeja's left-arm spin.
The tourists resumed on 207-2 in reply to India's 445 but lost two wickets in the space of six deliveries inside the first 30 minutes.
Jasprit Bumrah struck in the fifth over of the day when Root attempted a reverse scoop and Jaiswal caught it at second slip.
Kuldeep got Jonny Bairstow trapped lbw for a duck.
The left-handed Duckett, who smashed 23 fours and two sixes in his 151-ball knock, got to 153 but came to a tame end when he chased a wide delivery from Kuldeep to be caught at cover.
The five-match series is level at 1-1 after England won the opener and India bounced back in the second match.