The Women’s Tennis Association said Novotna, who had cancer, “died peacefully, surrounded by her family”.
The Czech player had lost in the Wimbledon final in 1993 and 1997 before winning the Grand Slam tournament in 1998 by beating Nathalie Tauziat.
Novotna captured the hearts of fans when she burst into tears after losing to German great Steffi Graf in 1993 and was consoled by the Duchess of Kent.
“Jana was an inspiration both on and off court to anyone who had the opportunity to know her,” said WTA chief executive Steve Simon.
“Her star will always shine brightly in the history of the WTA. Our condolences and our thoughts are with Jana’s family.”
Novotna was renowned for her serve-and-volley game, and achieved a career-high singles ranking of number two.
In addition to her only singles Grand Slam win at Wimbledon, she claimed 12 Grand Slam doubles titles and four in mixed doubles.
She was also inducted into the Tennis Hall of Fame in 2005.
It was Novotna’s exploits at Wimbledon which particularly endeared her to supporters, especially the 1993 defeat by Graf at the All England Club.
Novotna had a 4-1 lead in the third set and was a point away from going 5-1 up only to serve a double fault and lose five games in a row as she was beaten 7-6 (8-6) 1-6 6-4.
She started crying when presented with her trophy before the Duchess of Kent put a comforting arm around her and gave her a shoulder to shed her tears on during emotional scenes on Centre Court.
Novotna said the Duchess had told her “she would do it” when she went to collect her trophy and, despite losing to Martina Hingis in 1997, she finally won Wimbledon a year later.
In doing so, she became the then oldest first-time Grand Slam singles winner in the Open era at 29 years and nine months. (Courtesy BBC)