Portugal let a two-goal first-half lead slip as a second-half fightback from Serbia saw both sides share the spoils in a 2-2 draw.
The teams came into this clash on the back of victories, with Serbia having dispatched Ireland 3-2 on Wednesday, while Portugal narrowly saw off Azerbaijan on home turf.
The changes paid dividends as Jota headed home a perfect cross from Bernardo Silva to break the the deadlock in the 11th minute. Both sides went back-and-forth after the opening goal but it was Portugal and Jota who once again got on the scoresheet. The Liverpool forward scored his second header of the game, this time off a cross by Arsenal full-back Cedric. The European champions took a two-goal lead into the break and it seemed like they were coasting to another victory.
But Serbia had other plans and halved the deficit just 26 seconds into the second half. Halftime substitute Nemanja Radonjic whipped a dangerous cross into the box which Aleksandar Mitrovic rose highest to head home.
The hosts upped the ante in search of an equalizer and came close twice, first through Dusan Tadic and then through Sergej Milinkovic-Savic, but Portugal survived and maintained their lead.
The pressure from Serbia finally paid off as Filip Kostic slotted home a perfect through ball from Radondjic to level the scores. Both managers made several changes in the final 15 minutes as each went for the kill, but neither side was able to carve out a winner.
With just three minutes of additional time shown on the board, there was still ample time for two noteworthy incidents to take place.
First, Fiorentina defender Nikola Milenkovic was shown a straight red card for a rough tackle in the second minute of added time. Less than a minute later, Cristiano Ronaldo thought he had won it for Portugal when he beat Serbian goalkeeper Marko Dmitrovic from an acute angle. A retreating Stefan Mitrovic cleared the ball off the line but replays showed that the entire ball had crossed the goalline.
The Strasbourg defender also blocked a follow-up shot from Bernardo Silva on the goalline which was the last action of the game. Ronaldo was livid that the goal was not given but without VAR or goalline technology, the assistant referee's decision stood despite the protests from the Portugal captain.
An enraged Ronaldo was shown a yellow card for dissent but he continued his protests by walking off the field before the full-time whistle. It was an action-packed end to what was an exhilarating game and below we look at five talking points from the fixture.
#5 Diogo Jota shines brightly for Portugal
Diogo Jota was one of the stars of the Premier League season before a long-term injury halted his progress.
Having returned to full fitness last month, the 24-year-old was called up to the Portugal squad but only made it to the bench against Azerbaijan. He started against Serbia and rewarded his manager with two well-taken goals in the first half. The Liverpool forward showed great awareness and positioning to take advantage of opportunities in the box.
He now has five goals in 11 games for Portugal and is quickly becoming a mainstay in the front three.
#4 Aleksandar Mitrovic becomes Serbia's highest goalscorer
Over the last few years, Aleksandar Mitrovic has made a name for himself as one of the best target men in England and his goals have come in handy for Fulham and Newcastle United. The former Anderlecht man has struggled for form and playing time this season but despite his situation at club level, Mitrovic remains a crucial player for the Serbian national team.
Having started the game against Ireland on the bench, Mitrovic was introduced in the second half and scored a brace to power his side to a victory. This earned him a start against Portugal and he helped spark the comeback against the Iberians. His aerial ability came to the fore right at the start of the second half when he powered his way between three defenders to score the equalizer.
The Fulham striker has scored 29 goals in his last 31 games for Serbia and his latest, his 39th for Serbia, saw him become the country's all-time leading goalscorer, surpassing Stjepan Bobek's mark of 38 goals that he had set 65 years ago.
Speaking after the game, a tearful Mitrovic admitted to this being his dream come true.