It came as former Prime Minister Sir John Major warned that a Brexit vote is likely to result in another Scottish independence referendum which the Yes camp may win, leading to the “break up of the UK.”
First Minister Nicola Sturgeon has indicated that if the UK votes to leave the UK, while Scots vote to stay in, it would represent a “material change” in circumstance which would justify holding another referendum.
Senior nationalist MSPs have recently played down the prospect of another quickfire vote on leaving the UK in the event of Brexit, as polling evidence suggests there is no public support and that Scots would again vote to stay in the UK.
The Chancellor was joined on the EU campaign trail in the Borders yesterday by Scots Tory leader Ruth Davidson.
Mr Osborne said: “There are lots of things at risk in this referendum if we leave. Britain’s place in the EU and our stature as a nation is at risk. It is the Scottish nationalists themselves who said that they would use the Brexit vote as an excuse to bring up the issue again of Scottish Nationalism and separation.
“But the most recent Scottish elections made clear there wasn’t a mandate for that. I would say the simplest and easiest way to take the issue off the table is to vote for remain.”
Mr Osborne warned yesterday that a Brexit vote would result in more than 40,000 Scots job losses and a £4.5 billion economic hit.
Ms Davidson said a second referendum was not in the SNP’s recent manifesto and they didn’t win a majority in the Holyrood election, meaning a Remain vote would “kick away the final leg of a three-legged stool the SNP have” on the issue.
Sir John, who was speaking in Northern Ireland and appearing on a platform with former Labour prime minister Tony Blair, said quitting Europe against the wishes of Scots could make pressure for a second vote on Scottish independence “politically irresistible”.
Scotland previously voted by 55% to 45% in favour of remaining in the UK, with Sir John recalling how shortly before the vote “thousands and thousands of people gathered in Trafalgar Square to wave union jacks and sing songs” urging people to vote No to independence.
Sir John said: “Wisely, against the advice of the SNP, the Scots did just that.
“As a result with one qualification, the matter is now settled for a generation and beyond.
“But it is a very big qualification, because if Scotland votes in referendum to stay in EU, but the UK as a whole chooses to leave, there is a serious risk of a second referendum.
“Not straight away perhaps but ultimately, nationalist pressure for another shot at leaving the UK in Scotland could prove to be uncontrollable and politically irresistible. In those circumstances, if the UK were outside the EU, I can well envisage a different result in that referendum.”