Chris Ashton became latest experienced name to effectively see his World Cup dream end as he withdrew from the England squad for family reasons.
The Sale wing, 32, did not join the first tranche of players to attend an England training camp on Sunday and looks to have ended his chances of making the tournament in Japan.
Ashton's wife Melissa is about to give birth for the second time, so he has decided to support her rather than train with Eddie Jones' squad.
Chris Ashton has withdrawn from England's first pre-World Cup training camp
INITIAL 29-MAN TRAINING SQUAD
FORWARDS: Dan Cole (Leicester), Tom Curry (Sale), Alex Dombrandt (Harlequins), Tom Dunn (Bath), Charlie Ewels (Bath), Ellis Genge (Leicester), Joe Launchbury (Wasps), Lewis Ludlam (Northampton), Val Rapava- Ruskin (Gloucester), Nick Schonert (Worcester), Brad Shields (Wasps), Kyle Sinckler (Harlequins), Jack Singleton (Worcester), Sam Underhill (Bath), Mark Wilson (Newcastle).
BACKS: Chris Ashton (Sale), Mike Brown (Harlequins), Joe Cokanasiga (Bath), Elliot Daly (Saracens), George Ford (Leicester), Jonathan Joseph (Bath), Jonny May (Leicester), Ruaridh McConnochie (Bath), Dan Robson (Wasps), Marcus Smith (Harlequins), Ben Te’o (Worcester), Manu Tuilagi (Leicester), Anthony Watson (Bath), Ben Youngs (Leicester).
With Test veterans Chris Robshaw and Danny Care not selected by Jones, former captain Dylan Hartley still struggling with a knee injury and Jack Nowell out until late August after knee surgery England are already without high-profile names ahead of the tournament.
Ashton, who has won 44 caps, said: 'When I returned from Toulon last season it was for two reasons; firstly for my family and secondly to try and realise the dream of playing for England again.
'Pulling on the white shirt is always special and I'm grateful to Eddie for giving me the opportunity to have done so again.
'My wife, Melissa, is pregnant with our second child and I don't feel it is right for me to commit to being away in camp at this stage.
'In the meantime I wish Owen and the boys all the very best with their preparation over the coming months.'
A Barbarians hat-trick against England a year ago helped put Ashton back in contention
Just over a year ago Ashton was firmly out of contention as he played in France, having moved there after years of being disenfranchised by England.
However, after a record-breaking season where he scored 24 tries in the Top 14 season he returned to Twickenham for the Barbarians and scored a hat-trick against his country last May.
After that match Ashton held positive talks with Jones and then ripped up his Toulon contract to sign for Sale, for one last shot at the World Cup.
The winger played his first Test in four years last autumn against South Africa, started and scored against the All Blacks the following week then featured against Japan as well as Ireland and France in the recent Six Nations.
Injury curtailed the end of his club season, however he was picked in Jones' initial training squad, but now this decision has effectively ended his World Cup hopes.
Ashton opted to leave France and return to Sale last season to boost his World Cup hopes
Ashton would have been hard pressed to make the final 31-man party such is the competition in the back-three.
Elliot Daly, Jonny May, Joe Cokanasiga, Anthony Watson and Nowell are all sure to make the plane if fit, with Mike Brown, Ruaridh McConnochie and Ollie Thorley also vying for spots.
Ashton's withdrawal may well open the door for a late Brown return - the 33-year-old Quins man has not won a cap for a full 12 months but enjoyed a strong club season - or hand a chance for a 'bolter' pick such as former GB Sevens star McConnochie or 22-year-old Gloucester flyer Thorley.
Both of those latter two are yet to win a cap, so if either make the final squad Jones will be without highly-experienced men.
Having axed Robshaw and Care, seen Hartley - at this stage - ruled out with injury, and Ashton withdraw Jones may forego 291 caps-worth of experience.
Bath wing Ruaridh McConnochie only switched from Sevens only last summer