Southgate, who answered an SOS from the Football Association in the aftermath of Sam Allardyce's shock departure one game into his tenure, has worked with both players in his previous role as England Under-21 boss.
Rashford was a surprise omission from Allardyce's first squad in August, dropping down to Southgate's Young Lions and scoring a brilliant hat-trick against Norway on debut, while Lingard is in line for a first cap having been an unused substitute under Roy Hodgson last November.
The pair were named in a 23-strong party which will host Malta on Saturday before travelling to Slovenia three days later, with Glen Johnson, Ryan Bertrand and Alex Oxlade-Chamberlain also returning.
Johnson, 32, was last seen in an England shirt in the 2014 World Cup and may be surprised by the news despite a solid showing in Stoke's 1-1 draw at Old Trafford.
A knee injury to Nathaniel Clyne opened the door for him with Liverpool team-mate Adam Lallana, scorer of the only goal of the Allardyce era, and Leicester's Danny Drinkwater also ruled out this weekend, joining Harry Kane, Luke Shaw and Jack Butland on the absentee list.
Southgate, explaining his maiden selection, told FATV: "We've tried to keep some continuity from last month because my view would be a lot of things have been going right.
"Everybody's been focusing on the things that maybe can improve with the team but I think a lot of things have been going right. That continuity is important.
"We had a few people unavailable through injury, obviously Harry Kane, Jack Butland and we lost a couple over the weekend...Danny Drinkwater, Adam Lallana, Nathaniel Clyne and Luke Shaw's not quite back fully fit yet.
"That's had a bearing on the squad and a couple of players I've worked with, with Marcus Rashford and Jesse Lingard, in the Under-21s who we think are exciting players and we wanted to put in this squad."
Southgate offered only a gentle nod to the dramatic departure of Allardyce, whose contract was terminated after just 67 days in his dream job following remarks filmed by undercover reporters from the Daily Telegraph.
"It's obviously been a difficult week in a number of ways," he acknowledged.
"However we are in a position now where it's important to get the squad named. I'm looking forward now to working with the players because it's important that we build on the performance and the result against Slovakia."
Southgate will face the press for the first time at St George's Park on Monday morning, when he will expect to be quizzed further on the circumstances that led to his promotion for the next four matches at least.
Michail Antonio, who linked up with the national side for the first time last month, retains his place but there was no lifeline for Everton playmaker Ross Barkley or Jack Wilshere, who were both disappointed to be overlooked by Allardyce.
Wilshere left Arsenal for a loan spell at Bournemouth in a bid to restate his international credentials but is still looking to end a two-year wait for a full 90 minutes on the pitch.
Wayne Rooney, who has played alongside Southgate for England as their careers briefly intersected, was not formally named as captain but is understood to have been handed the armband.