Saints put up a strong fight against Bolton, but an Ivan Klasnic strike saw the hosts bow out of the Carling Cup at St. Mary's.
For the second consecutive season Saints pushed a Premier League side all the way in the second round stage, with the clinical manner of Birmingham's win last season mirrored in this tie.
Large parts of this game saw the home side dominated possession and chances, standing up to the physical challenge that Owen Coyle's men brought to the table.
The sole strike of the match arrived just past the half hour mark as Croatian forward Klasnic turned home Kevin Davies' pass inside the six yard box.
From there on Saints had the better of the chances to take the game into extra-time including a late flurry of corners in the last five minutes, but Kelvin Davis had to be alert to spectacularly deny Wanderers a second in the last minute as substitute Johan Elmander almost snapped up one of the few opportunities that the visitors had on goal.
Saints: Davis, Harding, Fonte, A. Martin, Butterfield (Mills 81), Lallana, Hammond, Schneiderlin (Chamberlain 75), Puncheon, Barnard, Lambert (Connolly 78). Subs (not used): Bialkowski, Jaidi, Dickson, Wotton.
Bolton: Bogdan, Taylor, Gardner (Holden 65), Knight, M. Davies, Ricketts, Alonso, O'Brien, Blake, Klasnic (Elmander 46), K. Davies. Subs (not used): Jaaskelainen, Petrov, Muamba, Cahill, Robinson.
Referee: Mr. K. Hill (Hertfordshire)
Attendance: 10,251 (including 460 away)
New signing Guly do Prado did not receive international clearance in time so Saints lined up the same from the side that began against Leyton Orient on Saturday minus Radhi Jaidi whose place in the centre of defence was taken by Aaron Martin.
The hosts kicked off and looked bright early on with Barnard and Lambert providing plenty of endeavour in attack, with the former getting the first shot of the game away at the near post which flew narrowly over the top of Adam Bogdan's goal.
Wanderers' central defensive duo Zat Knight and Andy O'Brien were given plenty to do early on as Saints poured forward, using every available attacking outlet, with Puncheon and Lallana also creating opportunities from the flanks.
On 18 it was Puncheon who drew a low save from the flame colour haired Bogdan, after collecting Barnard's throw-in from the right before unleashing a run and shot at goal from 30 yards.
Saints were awarded a free-kick in the 26th minute after Lambert was held as he battled to win an aerial ball. The resulting set piece from distance didn't put the striker off from trying his luck, but his effort flew wide of Bogdan's goal.
Moments later the visitors tested Kelvin Davis for the first time with midfielder Ricardo Gardner hitting a low drive through a crowd of bodies which the Saints shot stopper reacted well to, to keep the strike out.
At the other end, Barnard forced another fine save from Bogdan as the forward's low arrowed shot across goal was brilliantly kept out by the Trotters 'keeper.
The play quickly swung back up the other end where Bolton's slick movement in attack gave the visitors a slightly undeserved lead. Former Saint Kevin Davies controlled a cross and played the ball into the path of Ivan KLASNIC who prodded home from six yards out.
Harding was cautioned for a foul on Gardner ten minutes before the break and then five minutes alter Lallana followed him into the book after he was deemed to have taken a dive inside the area as he bore down on goal.
A cleverly worked short free-kick in the last minute of the half saw Puncheon and Lallana interchange with the latter finding space inside the area to shoot, but his curling effort just spun wide of the far post.
Saints continued to create inside the visitors' half right up until the whistle with optimism that the hosts weren't out of the tie just yet.
Half Time: Saints 0-1 Bolton Wanderers
At the start of the second 45 Wanderers replaced goalscorer Klasnic with Swedish forward Johan Elmander who partnered Davies in attack.
Saints started strongly again, with a fine passing move on 49 which involved Morgan Schneiderlin, Hammond and Lallana with the latter cutting the ball back across goal, but unfortunately no-one in a white shirt could get on the end of it.
On 54 Schneiderlin received the hosts' third booking of the evening for a late challenge on halfway, and Saints soon increased the ante once more in attack with Barnard and Puncheon making life difficult for Bolton's backline.
The same duo linked up to good effect just past the hour mark which resulted in Puncheon's shot being spilt by Bogdan and almost into the path of the striker, but the 'keeper managed to quickly gather the ball again at the second attempt.
The visitors saw Mark Davies booked for a foul on Schneiderlin before they made their second change of the match as Stuart Holden replaced Gardner in midfield.
A couple of minutes later Lambert eyed another free-kick from range which this time brushed the side netting on the far-side which had some supporters in the Itchen stand on their feet before the ball nestled behind the goal and out of play.
On 70 Robbie Blake whipped in a cross from the right which Kevin Davies connected to as he stuck his leg out, but the ball bounced off the striker's outstretched boot and into touch.
Danny Butterfield was the next to test Bogdan from distance, which the Trotters custodian had to turn out for a corner.
The first move from Alan Pardew saw Morgan Schneiderlin replaced by Alex Chamberlain for the last quarter of an hour as Saints pushed forward for an equaliser.
Another free-kick attempt from Lambert was the last piece of action from the Liverpudlian for the evening as David Connolly was introduced on 78, and then Joseph Mills got a run out as Saints withdrew Butterfield.
Kevin Davies was booked before Saints applied late pressure with four successive corners, and then Hammond was also cautioned as Wanderers tried to hit on the break.
In the 89th minute Davis made a superb reflex save to deny Elmander from close range and Harding was alert to clear the danger on the line before four minutes of additional time were added on for stoppages.
Connolly went the closest to an equaliser in that time as he saw his rising shot from inside the area narrowly clear the crossbar.
Davis came up for two last chance corners, the first of which Fonte headed wide, and the second the 'keeper just missed a connection to as the leveller Saints probably deserved never arrived.
Full Time: Saints 0-1 Bolton Wanderers
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