Juventus star Cristiano Ronaldo grabbed his 94th goal at international level in Portugal's 3-0 home victory over Luxembourg
England missed an opportunity to qualify for Euro 2020 Friday as they slid to a 2-1 loss in the Czech Republic, while Cristiano Ronaldo scored his 699th goal in a routine Portugal win over Luxembourg.
Harry Kane converted a fifth-minute penalty in Prague after Raheem Sterling was clipped by Lukas Masopust, the Tottenham Hotspur striker taking his qualifying tally to seven goals.
But Czech, humbled 5-0 by England in the reverse fixture last March, levelled shortly after as Jakub Brabec bundled in from a corner for his first international goal.
Substitute Zdenek Ondrasek swept home the winner on his debut for the host on 85 minutes, a goal that ended England's decade-long unbeaten run in qualifying spanning 43 matches.
England's last qualifying defeat came ahead of the 2010 World Cup when they lost 1-0 away to Ukraine in October 2009.
Southgate's side remain top of Group A on 12 points alongside the Czechs, who have played a game extra.
England could still clinch qualification Monday with victory in Bulgaria if Kosovo fail to beat Montenegro.
World champion France claimed a 1-0 win away to Iceland as Olivier Giroud delivered the only goal with a penalty on 66 minutes in Reykjavik.
Giroud has played sparingly for Chelsea this season but the 33-year old is now just four goals behind Michel Platini (41) for second on France's list of all-time leading scorers.
France are level on 18 points with Turkey atop Group H ahead of Monday's showdown in Paris, where victory would send Didier Deschamps' men through to the finals.
Cenk Tosun popped up with a 90th-minute winner for Turkey as they scraped past Albania 1-0 in Istanbul.
Iceland's first home loss in qualifying since June 2013 dropped the Nordic island nation six points adrift of the top two.
Juventus star Ronaldo grabbed his 94th goal at international level in Portugal's 3-0 home victory over Luxembourg.
Bernardo Silva opened the scoring in Lisbon in the first half before Ronaldo added a second on 65 minutes with a magnificent chip following an error in the visitor's defence.
Goncalo Guedes added a late third although defending champion Portugal still trail Group B leader Ukraine by five points.
Portugal next travel to Kiev to face a Ukraine side on the cusp of qualification following a 2-0 home win over Lithuania secured by Atalanta midfielder Ruslan Malinovskyi's brace.
A goalless draw between Montenegro and Bulgaria eliminated both teams from contention in Group A, while it was a night to remember for Andorra.
The minnow won their first European qualifier in history after 56 defeats in a row as Marc Vales' second-half header earned Andorra a 1-0 victory over 10-man Moldova.
Czech Rep 2 (Brabec 9, Ondrasek 85) England 1 (Kane 5-P)
Montenegro 0 Bulgaria 0
Portugal 3 (Bernardo 16, Ronaldo 65, Guedes 89) Luxembourg 0
Ukraine 2 (Malinovskyi 29, 58) Lithuania 0
Andorra 1 (Vales 63) Moldova 0
Iceland 0 France 1 (Giroud 66-P)
Turkey 1 (Tosun 90) Albania 0