Winning Europe's elite club competition had been a burning desire for Roman Abramovich since the Russian billionaire took over as the owner at Stamford Bridge in 2003.
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Di Matteo found the 'Holy Grail' against all the odds in May but Tuesday's 3-0 reverse at Juventus, a defeat that leaves Chelsea on the brink of Champions League elimination, prompted Abramovich to end the reign of his eighth manager at the club.
"The team's recent performances and results have not been good enough and the owner and board felt a change was necessary now to keep the club moving in the right direction as we head into a vitally important part of the season," the club said on their website.
"The club faces a difficult task ahead in qualifying for the knockout stages of the Champions League as well as maintaining a strong challenge for the top of the Premier League while competing in three other cup competitions," the club said.
"The owner and the board would like to thank Roberto for all he has done for the club since taking over in March. Roberto helped guide us to an historic Champions League victory and a seventh FA Cup.
"We will never forget the huge contribution he has made to this club's history and he will always be welcome at Stamford Bridge."
Chelsea then knocked out Benfica in the quarter-finals before beating overwhelming favourites Barcelona in the semis and defeating Bayern Munich on penalties in the final in the German team's own Allianz Arena stadium.
