Newspaper the Herald Scotland has alleged that VisitScotland's priorities were "controversial" as the payment covered the period when Lederer took over as chairman in 2001.
The payments to Gleneagles included more than £70,000 for hosting the Scotland Means Business event in 2002 and more than £54,000 for holding the US Tour Operators Conference in 2005.
However, Philip Riddle, chief executive of VisitScotland, said that decisions to hold events at Gleneagles were purely operational and not discussed with the VisitScotland board or the chairman.
"As chairman of VisitScotland Peter Lederer has never been involved with any decisions regarding expenditure at Gleneagles," he said.
"Gleneagles is one of the most iconic destinations in the world. For VisitScotland not to use it in the promotion of the country would be remiss."
Many of the events held at Gleneagles take place at different five-star resorts in Scotland on a rotational basis as they are aimed at showcasing the best accommodation and facilities the country has to offer.
Two of the events held at Gleneagles, highlighted by the Herald Scotland, have alone generated more than £5m for the Scottish economy.
Riddle added: "Lederer has been the most fantastic ambassador for Scotland and acts with the utmost integrity at all times.
"This is reflected in his standing within the industry and across the world. To imply otherwise is quite simply absurd."
Lederer, who was appointed chairman of Gleneagles in 2007, is due to step down as chairman of VisitScotland on 31 March 2010.
Peter Lederer re-elected as VisitScotland chairman >> By Janet Harmer
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