Sunday's 2-1 home defeat to struggling Villarreal, who had not won on the road this season, means Sevilla have not won a league game since early December.
"Sevilla took the decision to sack first-team coach Marcelino this morning due to the negative run of results that has seen them go seven matches without winning and collect only two points," the Andalucian side said in a statement.
With 26 points from 21 matches, Sevilla are four points above the relegation places and six adrift of the Champions League qualification berths.
Marcelino got off to a poor start with Sevilla after replacing Gregorio Manzano in June, when the UEFA Cup winners in 2006 and 2007 failed to make it past the first hurdle in the Europa League in August.
They were edged out of the King's Cup by Valencia in the quarter-finals last month.
No successor has yet been named, but local media were suggesting former Getafe boss Michel as a possible replacement.
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