The Japanese group played their way into the Guinness Book of World Records on Saturday at an event held in Yokohama, south-west of Tokyo.
Thousands of musicians, of all ages, headed to the Japanese capital where they performed together to set the new world record.
The ensemble entertained world record officials with an interpretation of Aloha Mahalo A Hui Ho, a song written by Hawaiian-born former sumo wrestler Konishiki.
Konishiki, who retired from wrestling in1997 as then heaviest sumo of all-time, joined the ensemble on stage to also become a world record breaker.
Consisting of 2,134 people, the world record eclipsed a record set in Sweden last year consisting of 1,547 players.
The Japanese performance was part of Ukulele Picnic Week which also featured hula dancing.
Largely associated with Hawaii, the string instrument was introduced to The Aloha State by Portuguese immigrants in early 1880s.
It reached Japan by 1929, where it quickly gained popularity as enthusiasm for western music grew in the years leading up to World War II.