Real Betis were deserving victors of a thrilling contest at the Benito Villamarin where two galling Real Madrid errors cost them their place at the top of La Liga, with Barcelona now two points clear.
Real Madrid's nightmare afternoon was made worse by another injury to Eden Hazard, who was forced off in the second half of their 0-1 defeat to Levante.
Barcelona can cut the gap back to three points by beating Levante at home on Sunday but Atletico Madrid now trail leaders Real Madrid by 13 points.
Karim Benzema scored a dramatic injury-time equaliser to salvage a 1-1 draw for Real Madrid against Valencia at Mestalla.
Real Madrid are a point ahead of promoted Granada and Atletico Madrid after Diego Simeone's side failed to break down their well-drilled local rivals.
La Liga: Real Madrid are now four points clear of Barcelona after five games played.
Karim Benzema's brace helped Real Madrid secure three points at home against Levante.
Zinedine Zidane said he believes Gareth Bale will now stay, with the Spanish transfer window closing on September 2.
Real Madrid's response to Barcelona winning the title on Saturday night was to slip to a 0-1 defeat at Vallecas and fall 18 points behind La Liga's newly-crowned champions.
Real Madrid were mathematically ruled out of the title race after Barcelona beat Alaves on Tuesday while Atletico Madrid look likely to finish above them too for a second consecutive year.
Karim Benzema has been one of the few bright sparks in an otherwise dreary campaign for Real Madrid and his hat-trick at the Santiago Bernabeu took his tally to 30 goals for the season.
Real Madrid delivered another thoroughly underwhelming performance at Butarque to remain 13 points adrift of Barcelona.
Gareth Bale departed again to whistles with 14 minutes left, a conciliatory hand slap from Zinedine Zidane offering little consolation as he trudged off.
Real Madrid still lag 12 points behind leaders Barcelona, who are 10 clear of Atletico Madrid and heavy favourites to retain their La Liga title.
Santiago Solari was speaking at a press conference ahead of Tuesday's meeting with Ajax at the Bernabeu in the Champions League.
Karim Benzema and Vinicius Junior each scored in a 3-0 win over Alaves on Sunday.
Gareth Bale had been out since January 3 with a calf strain but made an instant impact off the bench for Real Madrid.
Karim Benzema, who has not played for France since being left out of the squad ahead of Euro 2016, has been dogged by controversy in previous years.