Scott Meikle, owner of the five-room Moor of Rannoch Hotel near Pitlochry, said five customers had already contacted him this morning to cancel their bookings – while others had asked to move their breaks to later in the year.
Meanwhile, Gethin Chamberlain, owner at self-catered accommodation The Five Turrets in Selkirk, said he had taken his first cancellation while Nicola Sturgeon was still giving her speech in Parliament on Tuesday, when the First Minister unveiled her route map out of lockdown.
Ms Sturgeon said in the last week of April, Scotland could expect to see “a phased but significant” re-opening of the economy.
However, tourism businesses have warned uncertainty surrounding details of who will be able to travel and when will leave them unable to guarantee bookings – compared to businesses in England, where Boris Johnson has set dates on lockdown easing.
Mr Meikle, who runs the hotel with his wife Steph, said: “We have had some guests cancelling already and some guests moving bookings to later this year. We also have a few group bookings in and our worry is that if the two household rule applies, they will also have to cancel.
"Our biggest issue is if we move back into the levels system with travel restrictions. Even last week, we were getting quite a few new bookings, but now, with the uncertainty, it has just dried up.”
Mr Meikle said the Scottish Government grant of £2,000 a month did not even cover half of the £5,500 a month it costs to keep the hotel building running without any guests.
Mr Chamberlain said he believed many tourists would shy away from booking Scottish hotels and said the tight restrictions have "really serious implications” for Scottish businesses.
He said: "We took our first cancellation while Sturgeon was still on her feet yesterday and we continued to take cancellations during the day. People will look at England and see that it gives them the prospect of a holiday – they will have confidence in it.
"Whereas in Scotland, all they can see is nothing because there's nothing they can hold onto. Who's going to book a holiday with us? How do we compete?
“We've got wide open spaces in the borders and it's a perfect place to come for a holiday, and yet the only people who can realistically visit in the near future are people who already live here. It's soul-destroying."
Speaking at Wednesday’s coronavirus briefing, Ms Sturgeon said: "I’ve tried to be straight with people. I didn’t say to any business that it would be safe to start taking bookings, though it will be soon.
"But I don’t want businesses to be in a position where I’m giving them a date that I can’t back up and then go back on that. As we go through the next weeks, we will add to the clarity we have been able to give as much as we possibly can.
"I’m sorry for any business in the situation you are just describing. I don’t want a single business to be living with these restrictions longer than is necessary.”