The organisers of the Etape Caledonia hope it will claim a prominent place on the world cycle-racing calendar.
The inaugural event, on June 24, will feature more than 2000 metres of hill climbs in the stunning surroundings of Highland Perthshire. Organisers say the 83-mile course will provide a "stern but achievable challenge" for up to 2500 competitors.
But not half as stern as the challenge they are facing from the congregation of Dull and Weem Church of Scotland.
Elders say the five-hour closure of the B846 along the Tay near Aberfeldy will cut them off from their Sunday service.
They are threatening legal action against Perth and Kinross Council, which is backing the first annual Etape Caledonia event.
The competition - strictly speaking not a race, as the participants test themselves against the clock - is expected to generate more than £500,000 for the local economy.
But Tom Pringle, an elder at the church, just two miles outside Aberfeldy, said the road closure would breach his and other families' right to worship.
He said: "It was with disgust that I heard this cycle event had been agreed, along with the necessary closure of many roads in Highland Perthshire.
"All this seems to have been done minus any consultation with the people living, working and worshipping in the area."
Councillor John Hulbert, vice-convener of Perth and Kinross Council's Enterprise and Infrastructure Committee, which agreed to the race taking place, said he would attempt to find a "compromise" for concerned worshippers but did not know if one could be found.
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