The woman held her left hand in the air and flipped off the commander-in-chief while the luxury fleet passed her by.
Trump had just left his private golf club near Washington, DC, when the woman was pictured giving him a one finger salute.
She gave him the finger again when she caught up to the motorcade at a traffic light, while another woman gave him a “thumbs-down sign”.
Protesters – including a woman with a sign reading “Impeach” – had gathered outside the club as the business mogul-turned-president spent a few hours playing golf.
A White House pool report detailed the incident near Trump National Golf Club in Sterling, Virginia, about 20 miles from the Oval Office.
It stated: “POTUS’s motorcade departed the Trump National Golf Club at 3.12pm, passing two pedestrians, one of whom gave a thumbs-down sign.
“Then it overtook a female cyclist, wearing a white top and cycling helmet, who responded by giving the middle finger.
“The motorcade had to slow and the cyclist caught up, still offering the finger, before turning off in a different direction. Motorcade is now gathering speed and heading for DC.”
It was unclear whether the president saw the woman’s gesture.
The image was captured by photographer Brendan Smialowski, and it soon went viral as Trump’s critics celebrated the unidentified woman.
Trump – who criticised former president Barack Obama for spending “too much time” at golf clubs – had little support in the Sterling area during last year’s presidential election.
More than half of voters (55 per cent) supported Democratic challenger Hillary Clinton, with 38 per cent voting for Trump.
Trump has faced criticism over the amount of time he has spent at his golf clubs since taking office in January.
MSNBC producer Kyle Griffin wrote on Twitter that the visit to the golf club on the Potomac River was Trump’s 96th day at one of his properties as president.
Seventy-six of those visits have been to golf clubs, Newsweek reported.
Meanwhile Trump’s approval rating has hit a new low, dipping to 38 per cent just ten months into his four-year term, a new NBC News/Wall Street Journal poll revealed.