Theresa May said voters should choose the Conservatives in the local government ballot to send a “clear message” on the constitution.
Ever since the UK voted to leave the European Union but Scotland voted to stay in, there has been speculation that the Scottish Government will seek to hold another referendum.
The First Minister has warned that Mrs May’s plans for a hard Brexit, taking the UK out of the single market, “undoubtedly” made that a more likely prospect.
But writing in Holyrood magazine, the Prime Minister said her party would be “looking forward to the local elections in May, when voters across Scotland will have the chance to send a clear message to the SNP that they do not want a second independence referendum, by voting Scottish Conservative and Unionist”.
Meanwhile, Scottish Labour has launched a dedicated website TogetherStronger.scot, where people can register their objection to a possible second independence referendum.
An SNP spokesman said: “There is already a cast-iron democratic mandate for an independence referendum - that was delivered in last year’s Holyrood election, however much the Tories might try to deny it.
“That mandate also stems from the EU referendum, which saw Scotland vote by a 24-point margin to stay in Europe - and Theresa May’s reckless pursuit of an economically ruinous hard Brexit will only strengthen opinion in Scotland against leaving Europe.
“The Prime Minister couldn’t be more wrong to suggest there is considerable common ground between her Government and the Scottish Government on Brexit - her party is hell-bent on taking us out of the world’s biggest single market, with all the economic damage that would cause, while we are intent on protecting Scotland’s vital national interests.”