England have not beaten the Springboks in 10 Tests dating back to the autumn of 2006 and go into this encounter fuelled by the hurt of two near misses.
Rowntree felt England should have beaten Australia last week and they might also have defeated the Springboks in the third Test of the summer tour, which ended in a 14-14 draw in Port Elizabeth.
"We are still hurting from last week. We lost and we are not happy about it. It was a game we should have won," said Rowntree, England's forwards coach.
"That last game in Port Elizabeth was also a game we should have won.
"We had lost two Tests on that tour. In that last hour before the game in Port Elizabeth we decided that wasn't going to happen again.
"And we have to recreate that atmosphere tomorrow [Saturday], that pure desire for it not to be a defeat again.
"You bank up defeats and frustrations and we seem to have done that over the last half-dozen games - and that has got to come out in a positive performance."
England's win against Fiji on the opening weekend of the QBE autumn internationals was their only victory in five Tests since the end of the RBS 6 Nations championship.
Head coach Stuart Lancaster said at the outset of the autumn series that the time had come for England to deliver against the might of the southern hemisphere.
He had called it, perhaps a touch prematurely for a young team, "production time".
England had their chances to win last week and should have taken them but for long periods they were outplayed by an Australia side that were too smart and too savvy.
The South African challenge will be different and England know what to expect.
The Springbok game is based on ferocious forward power, with Ruan Pienaar and Patrick Lambie pulling the strings in behind.
The challenge for England is dealing with it. They failed in the first two Tests of the summer tour but Port Elizabeth is their reference point.
England ratcheted up the ferocity in that final Test and went toe-to-toe with the Springboks to end a run of nine straight defeats in the fixture.
"The mentality we achieved in the hour before the game got us in the right place," Lancaster recalled.
"You have to make sure you are not so revved up that that you are a bull in a china shop and can't think clearly.
"We've got to hit the balance between being on the edge, absolutely on the edge, but also having clarity of mind."
That, arguably, is what cost England last week when captain Chris Robshaw opted against kicking for goal in favour of lineouts and quick tap penalties as they chased the game.
England felt the Wallabies were at breaking point but Thomas Waldrom dropped the ball over the line and Chris Ashton was tackled just short as Australia held on.
"I bloody hate hearing these coaches on the back of a defeat saying 'we're learning' but the fact is we are still a very young group," Rowntree said.
"What really pleases me is that at the end of every game, we're in it, we're forcing the pace.
"We haven't beaten this nation for a while. We've got them again, at Twickenham tomorrow [Saturday], on the back of a really frustrating defeat. There's a lot of energy there for us to thrive upon."
Rowntree's fighting talk continued the theme set earlier in the week by Tom Wood, who returns to the England team as one of six changes from the 20-14 defeat to Australia.
Wood, Alex Corbisiero, Joe Launchbury and Ben Morgan have been charged with bringing steel to an England pack that was second best to the Wallabies for an hour.
"James Haskell says that playing South Africans is like a man test," said Wood.
"They look at you, they look for your biggest guy and get theirs to run at him as hard as he can to see who comes out on top.
"If you do show any fear and if you do back off to any extent you've got a long day at the office. You have to match it head on."
The forecast is for wind and rain at Twickenham, which gives further credence to the selection of Mike Brown on the wing as England reprise the summer tour tactic of using two full-backs.
Brown, who has replaced specialist finisher Charlie Sharples, is a rock under the high ball and he offers a long-range left-footed kicking option.
"There will probably be a lot of set pieces, a lot of kicking," Rowntree said.
"The kick receipts and the lineout will be interesting with the wind. We need to be switched on."