Left-handed opener Shikhar Dhawan, who last played a 50-over match for India in June 2011, got the nod ahead of Gambhir while wicket-keeper batsman Dinesh Karthik, who can also open the batting, was recalled after almost three years.
The other opener in the squad, Murali Vijay, was a surprise inclusion as the right-hander last played an ODI against South Africa in Jan. 2011 and has failed to score a fifty in his 11 matches in the format.
Yuvraj, who made a fairytale comeback last year after battling a rare form of lung cancer, could not find a place in the Indian middle-order with selectors preferring right-handed batsman Rohit Sharma ahead of him.
Fast bowler Umesh Yadav returned from his back injury while all-rounder Irfan Pathan also got a recall. Off-spinner Ravichandran Ashwin and leg-spinner Amit Mishra were the two specialist slow bowlers in the squad.
India, the reigning 50-over world champions, are in Group B and will play the opening match of the June 6-23 tournament against South Africa in Cardiff. Pakistan and West Indies are the other teams in the group.
India squad: Mahendra Singh Dhoni (captain), Murali Vijay, Shikhar Dhawan, Dinesh Karthik, Virat Kohli, Suresh Raina, Rohit Sharma, Ravindra Jadeja, Ravichandran Ashwin, Amit Mishra, Irfan Pathan, Umesh Yadav, Bhuvneshwar Kumar, Ishant Sharma, R. Vinay Kumar.