Woods is missing after having back surgery, but the world number one has not won at Augusta since 2005 and the tournament is considered wide open.
Northern Ireland's McIlroy is the bookmakers' favourite to clinch a first Masters Green Jacket and third major.
Adam Scott bids to become the fourth player to win back-to-back titles.
The 24-year-old McIlroy suffered an infamous meltdown at Augusta in 2011 when he blew a four-shot lead going into the final round, but he has won two majors since and has put behind him a miserable 2013 season.
"Mind, body, equipment, it's all there," he said. "There are no excuses if I don't do well this week."
American left-hander Mickelson is the crowd favourite and can join Arnold Palmer and Woods on four Green Jackets, 10 years after winning his first.
"It's a magical place," said Mickelson. "I just love everything about this tournament."
The 43-year-old won the Open at Muirfield last summer for a fifth major title and the third leg of a career Grand Slam of all four majors, with only the US Open, in which he has a record six runner-up spots, missing.
Mickelson admits the Masters feels "weird" without Woods, who is missing the tournament for the first time since his debut as an amateur in 1995.
The 38-year-old has been stranded on 14 majors since 2008 as he chases Nicklaus's record of 18 major titles.
"I hope he gets back soon," said Mickelson. "Look what he's done for the game the last 17 years. It's been incredible.
"And no-one has benefited more from having Tiger in the game than myself."
Defending champion Scott is number two in the world and can overhaul Woods in the rankings with a two-way tie for third or better.
A second major title this week would make the Australian the fourth player to achieve back-to-back Masters wins after Woods in 2001/2002, Nick Faldo in 1989/1990 and Nicklaus in 1965/1966.
Another Australian Jason Day, who is fourth in the world, can also top the rankings if he wins. Day was second on his Masters debut in 2011 and third in 2013. Sweden's Henrik Stenson is the third player who can take Woods's crown if he finishes joint second with only one other player or better.
Britain is still searching for a first Masters champion since Faldo's third Green Jacket in 1996. The last European to win the Masters was Spain's Jose Maria Olazabal, who won his second title in 1999.
England's Lee Westwood finished runner-up in 2010 and tied for third in 2012, while countryman Justin Rose is the reigning US Open champion and has had two top 10s in his last six Masters.
"Knowing you've won a major and knowing you've faced those emotions and come through is a huge benefit this week," said Rose. "I'm coming in with no hype, a little under the radar, and I feel good."
A total of 24 players will make their Masters debut, with many suggesting the tournament could see only its third first-time winner after Gene Sarazen in the second event in 1935 and Fuzzy Zoeller in 1979.
America has a number of rising stars playing at Augusta for the first time, including Patrick Reed, 23, who has won twice this term to make it three PGA Tour titles in only his second season on the circuit. Compatriot Jordan Spieth, 20, also has a PGA Tour win under his belt and was named rookie of the year in 2013.
Reed and Spieth will partner McIlroy for the first two rounds, starting at 15:52 BST (10:52 local time) on Thursday.
Scott will tee off with 19-year-old English amateur Matthew Fitzpatrick and America's US PGA champion Jason Dufner in the group ahead, while Mickelson will play in the penultimate group with Rose and South Africa's Ernie Els at 18:48 (13:48).
This year marks the 50th anniversary of Arnold Palmer's fourth Masters win and seventh and final major title.
Palmer, 84, six-time champion Nicklaus, 74, and three-time winner Gary Player, 78, will get the tournament under way as ceremonial starters at 12:40 (07:40) on Thursday.
Thunderstorms caused Monday's practice round to be cancelled after just a few hours but Tuesday and Wednesday went ahead as planned and the forecast for the tournament days is sunny and warm.
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