Here, the PA news agency takes a look at five of the main talking points ahead of the Ahmad Bin Ali Stadium encounter.
Turning tide of history
Wales have not beaten England since May 1984 when the debut-making Mark Hughes sealed a 1-0 win in the British Home Championship. England have won the last six encounters, with their last competitive victory being a last-gasp 2-1 success at Euro 2016.
The odds appear stacked against Wales achieving the four-goal win they need to progress to the knockout phase – they have never beaten England by such a margin in the 103 games between the two countries. Failing that, they need to win and hope for a draw between the United States and Iran.
Kane able to go again?
“It was a problem of tiredness – really, really tired” – the alarming comments made by Antonio Conte just two and a half weeks ago after Harry Kane underwhelmed as Tottenham exited the Carabao Cup.
Questions over the 2018 Golden Boot winner’s freshness have continued onto Qatar, where he has yet to find the net, with England greats Wayne Rooney and Alan Shearer suggesting the record-chasing goalscorer should be rested against Wales.
The 29-year-old also had an injury scare after receiving a kick to his foot in the opener against Iran, meaning Southgate’s selection decision will be interesting.
Can Bale and Ramsey inspire Wales?
Gareth Bale and Aaron Ramsey have failed to make the impact they had hoped to at this World Cup.
Skipper Bale did win and convert the penalty to secure a 1-1 draw against the United States, but both players have fallen well short of their own high standards.
The pair have pushed Wales to dizzy heights over the last decade or so, and Dragons fans are hoping the two 30-somethings have one big game left in them.
Time to unleash Foden?
On the eve of the tournament, precociously talented Phil Foden admitted he has yet to translate his Manchester City form onto the international stage, having only scored twice in his 18 appearances until that point.
Foden was tipped to make a big impact in Qatar but has so far managed just 19 minutes off the bench with the Iran opener already won.
Southgate’s decision to leave him on the bench in the USA stalemate caused consternation and criticism of the England boss, who said he has a “big part” to play amid the push for the 22-year-old to start.
Rest players or push for top spot?
With the debate over Kane and clamour for Foden raging, there are other decisions to make as Southgate looks to balance the desire to finish top with keeping the squad fresh.
Jordan Henderson replaced teenager Jude Bellingham against the USA and could well make his first competitive England start in a year.
Kyle Walker and Kalvin Phillips need minutes after returning from injuries that threatened their World Cup places, while Trent Alexander-Arnold offers an exciting option at right-back.
It would be out of character for Southgate to make wholesale changes but the need to keep players engaged and fresh means some alterations are likely having used the same line-up for the first two games.