Upton connected on the first pitch he saw from Baltimore's Luis Ayala for a soft blooper to left field that fell in for a base hit.
Rich Thompson went in to pinch-run for Upton, who received another standing ovation as he walked off the field.
After the game, Upton gave his jersey to the lucky fan who'd won the "Shirts Off Our Backs" contest before receiving more cheers from the crowd, causing him to tear up as he tossed his hat into the seats.
"No," Upton said when asked if he was ever this emotional on a baseball field. "I tried to hold it as long as I could, but I just couldn't."
Wednesday's game -- a 4-1 win over the Orioles -- may have been the last for Upton in a Tampa Bay uniform, as his contract is expiring and he has a chance to test the free-agent market in the offseason.
"This is all I know," said Upton, who also teared up in the dugout after leaving the game. "I might not be back. Ten years with the team, I don't know, a lot of guys don't get to do that, and I've had that opportunity to be around great people -- people who care about one another. If it has to happen, I'm definitely going to miss them."
Before the game, Upton said that he hadn't really thought about this possibly being his last game with the organization. He would be open to re-signing, but admitted, "I have to do what's best for me."
"I guess we'll see which teams are out there and who's interested," he said. "If it is the Rays, then great. If they don't give me that opportunity, then obviously I have to make a decision."
Upton was taken by the then-Devil Rays as the first overall selection in the 2002 Draft, and he signed in September of that year. He made his big league debut in 2004 as an infielder, but he never found his niche defensively over the 45 games in which he appeared.
He didn't play in the Majors in 2005, but in 2006, Joe Maddon's first season as manager, Upton returned, playing third base in 50 games and continuing to struggle in the field.
Upton began the next season starting at second base, but halfway through the season, he was switched to center field, now his primary position, and he gradually gained the reputation of being one of the better center fielders in the game.
In 2008 he was part of the American League champion squad, who had just changed their image, and Upton's as well.
"To go from worst to first and see this organization turn things around and become a winning ballclub," Upton cited as his favorite memory. "Going to the World Series and winning the American League East. ... I'd have to say that '08 season."
Over the next four seasons, Upton was a part of two more postseason teams.
"I've spoken often this year about how much I think B.J. has matured as a baseball player," Maddon said. "He's matured as a person, of course, but as a baseball player, our relationship, how it's worked in the dugout, watching him play the game, all that stuff that's going on there -- all that stuff has dramatically improved throughout the course of this season."
Ben Zobrist, who has played with Upton for the last seven years, thinks his best is yet to come.
"I hope the Rays find a way to keep him and we can have him in the clubhouse more and see him continue to develop as a player," Zobrist said. "I think he's going to continue to just get better. There's no question seeing him this year that he's just got better."
Though he finished the season with just a .246 batting average, he had a career-high 28 homers, led the team in RBIs (78) and swiped 31 bases.
"It happens pretty fast," he said of his time with the Rays. "It seems like it was yesterday."
Evan Longoria, a teammate of Upton's since 2008, has had his ups and downs with Upton, but appreciates what he's done for both the organization and the community.
"Obviously, I hope that he's back," Longoria said. "I don't know how real that possibility is. ... I enjoyed playing with him for the five years that I did. He's always been a great teammate to me."
Maddon said that what he'll remember most about Upton should he not return is how much the 28-year-old has grown as both a player and a person.
"I really enjoyed our relationship a lot," Maddon said. "This year was truly the year I saw him blossom."
Greg Zeck is an associate reporter for MLB.com. This story was not subject to the approval of Major League Baseball or its clubs.