A 2-0 victory at fellow Saudi side Al Ittihad saw them surpass the previous record of 27 straight wins by a top-flight team, set by Welsh team The New Saints in 2016.
The 39-year-old Portuguese was the only Al Nassr player to score in the shootout as his Saudi side went out 3-1 on penalties, the tie having ended 4-4 on aggregate.
Nassr, who won last week's first leg 1-0 thanks to a Ronaldo goal nine minutes from time, were the better side at Al Awwal Park and took the lead on 17 minutes through Portuguese winger Otavio.
Cristiano Ronaldo scored his 21st goal of the season and his fourth in the Asian Champions League on Wednesday as his moment of magic proved decisive in Al Nassr beating Al Fayha 1-0 in an all-Saudi last 16 tie.
Former Liverpool striker Sadio Mane and Anderson Talisca also scored for the Saudi side late Tuesday as they made it three wins from three in the group stages of the premier Asian club competition.
The former Paris Saint-Germain forward, signed in August for a reported 90 million euros ($94 million), grabbed the second in the record Asian champions' win at the Azadi Stadium
Senin Sebai scored on the break to put the club from Dushanbe in front with a minute left in the first half and Istiklol held out under relentless pressure until Ronaldo levelled with a clever clipped finish in the 66th minute.
Cristiano Ronaldo enjoyed a winning debut in the Asian Champions League as he helped Al Nassr to a 2-0 victory against 10-man Persepolis in Iran on Tuesday night.
Neymar, signed last month from Paris Saint-Germain for a reported €90 million, was making his first start for the record Asian champions having debuted on Friday as a substitute in the Saudi Pro League victory at home to Al Riyadh.
A 2-0 victory at fellow Saudi side Al Ittihad saw them surpass the previous record of 27 straight wins by a top-flight team, set by Welsh team The New Saints in 2016.
The 39-year-old Portuguese was the only Al Nassr player to score in the shootout as his Saudi side went out 3-1 on penalties, the tie having ended 4-4 on aggregate.
Nassr, who won last week's first leg 1-0 thanks to a Ronaldo goal nine minutes from time, were the better side at Al Awwal Park and took the lead on 17 minutes through Portuguese winger Otavio.
Cristiano Ronaldo scored his 21st goal of the season and his fourth in the Asian Champions League on Wednesday as his moment of magic proved decisive in Al Nassr beating Al Fayha 1-0 in an all-Saudi last 16 tie.
Former Liverpool striker Sadio Mane and Anderson Talisca also scored for the Saudi side late Tuesday as they made it three wins from three in the group stages of the premier Asian club competition.
The former Paris Saint-Germain forward, signed in August for a reported 90 million euros ($94 million), grabbed the second in the record Asian champions' win at the Azadi Stadium
Senin Sebai scored on the break to put the club from Dushanbe in front with a minute left in the first half and Istiklol held out under relentless pressure until Ronaldo levelled with a clever clipped finish in the 66th minute.
Cristiano Ronaldo enjoyed a winning debut in the Asian Champions League as he helped Al Nassr to a 2-0 victory against 10-man Persepolis in Iran on Tuesday night.
Neymar, signed last month from Paris Saint-Germain for a reported €90 million, was making his first start for the record Asian champions having debuted on Friday as a substitute in the Saudi Pro League victory at home to Al Riyadh.
The Saudi Pro League side came from behind to triumph 4-2 in the play-off at Mrsool Park, with Brazilian Anderson Talisca scoring twice and Croatia international Marcelo Brozovic rounding off the win. Nassr had been two minutes from exiting the tournament