England who sent back Gambhir (4), Virender Sehwag (30) -- in his 100th Test – Sachin Tendulkar (8), Virat Kohli (19) and Yuvraj Singh (0).
Barring Gambhir and Yuvraj, who fell to Anderson and Graeme Swann, the others were dismissed by Panesar.
Pujara and captain Mahendra Singh Dhoni, who mixed caution with aggression before becoming Panesar's fourth victim for 29, put on a partnership of 50 runs in 128 balls.
England's hopes of slicing through the Indian tail, though, came to naught with Ashwin scoring his second fifty.
With some more batting to come in the form of Harbhajan Singh -- drafted into the eleven as the third spinner after more than a year's gap -- Zaheer Khan and Pragyan Ojha, India would fancy their chances of taking the score past 300.
In the morning and soon after lunch, England applied the screws on India and made the hosts struggle on a turning track on which the ball gripped the surface and also bounced.
Panesar, who was left out of the first Test, was the destroyer-in-chief, with a splendid spell of 23 overs on either side of lunch for the wickets of Sehwag, Tendulkar and Kohli before coming on for another spell for the wicket of Dhoni.
Anderson and off-spinner Swann packed off Gambhir (4) and Yuvraj (2) in two balls apiece.
Gambhir, who flicked Anderson to midwicket off the first ball, missed the next to be rapped in front.
After pushing India on the back foot in the pre-lunch session, England continued the good work with a two-wicket burst in successive overs in the first hour of the post-lunch session.
Panesar, who sent back Sehwag and Tendulkar in successive overs before lunch, got his third victim by dismissing Kohli.
After disturbing the stumps of Sehwag and Tendulkar with sharply turning balls, Panesar induced Kohli to attempt a drive with a ball that drifted in and turned big on pitching to take the leading edge for Nick Compton to bring off a good diving catch.
Kohli had cover-driven the left-arm bowler and his spin partner Swann for attractive boundaries in successive overs before he was lured by the crafty Sikh.
The slow bowler made the ball drift in to the right-handers and extracted sharp turn off the track.
He could have had the confident-looking Pujara too when the batsman, batting on 60 in a team score of 135 for five, edged the bowler to the left of Anderson who could not latch