Guthrie finished fifth in the Shriners Hospitals for Children Open in Las Vegas on Sunday after a closing 64 and carried on where he left off at Lake Malaren in Shanghai.
The 23-year-old, who only turned professional last year, carded birdies at the second, third and seventh to reach the turn in 33, before also picking up shots on the 13th and 14th and chipping in for an eagle on the par-five 15th.
After saving par on the final three holes, Guthrie signed for a flawless seven-under-par 65 to lead by three shots from former Open champion John Daly, with Peter Uihlein returning a 69 to ensure the only Americans in the field occupied the top three places.
"It was a great day," said Guthrie, who was one of only 13 players to break par in the windy conditions.
"It was one of the better rounds I've ever played. It's playing tough out there, it's hard to control the ball exactly where you want to leave it and today when I did miss I gave myself a chance and I was able to short-game my way around.
"I was really looking forward to this tournament on my schedule and I just can't be more happy to be here. I wanted to come over here, to get used to playing kind of a global schedule.
"I was able to stay up on my 12-hour flight and crashed at about 9pm here on Tuesday night - that was about as quick as I could get here from Las Vegas - so I'm actually on decent Shanghai time right now."
Daly's score was as remarkable as his garish red, black and yellow trousers - which he described as "ketchup and mustard" - given that it was his first competitive round since July.
The 47-year-old underwent elbow surgery after pulling out of the Greenbrier Classic after three holes of his second round but has returned to action a few weeks earlier than expected thanks to icing his arm and a diet of chocolate milk.
"I've always been a quick healer. I don't know why," Daly said.
"He (his doctor) figured four months. I did everything he told me to do. I usually don't listen to anybody."
This week's event is the first of four tournaments which make up the inaugural Final Series on the European Tour, with Sweden's Henrik Stenson holding a lead of 425,573 euros (£362,660) over Graeme McDowell at the top of the Race to Dubai standings.
But with a first prize of 850,000 euros (£725,000) on offer here and a total prize fund of almost £20million in the four events, the battle to become European number one in Dubai in mid-November is far from over.
Stenson, whose victory in the Tour Championship in Atlanta last month secured a £6.2million bonus for winning the FedEx Cup, carded a level-par 72 while McDowell produced a moment of genius in his 70.
The former US Open champion's approach to the par-five 13th came up just short of the green and left him 30 yards from the pin, but he duly holed the monster putt for an unlikely eagle.
Playing partner and fellow Northern Irishman Rory McIlroy had been two under with three holes to play but failed to get up and down from a greenside bunker on the 16th and had to settle for a 71.
The former world number one, now sixth in the rankings, has not won a tournament in 2013 after five victories in 2012 but was second in Korea last week and at least recorded the only birdie of the entire field on the 470-yard ninth.
The field had been reduced to 77 players by the withdrawal of Holland's Joost Luiten - twice a winner on the European Tour this season - due to a shoulder injury.
Luiten hit his opening tee shot before withdrawing, much to the disappointment of first reserve Justin Walters who was standing nearby.
Tour regulations state players must compete in two of the first three Final Series events to be eligible for the season-ending DP World Tour Championship. By hitting a single shot Luiten is deemed to have taken part and could therefore repeat the same trick in the HSBC Champions or Turkish Open to claim his place - and potential financial bonus - in Dubai.