Spain’s acting prime minister said on Wednesday that Spain is opposed to any EU discussion with Scotland after the referendum result.
"The Spanish government is opposed to any negotiations with anyone else but the British government," Mariano Rajoy said.
"The United Kingdom leaves and with it, all those who make up the United Kingdom," he said.
Spain has always been against independence for Scotland. Spain has its own areas which would like to leave, not least wealthy Catalonia, and is fearful of any precedent being set.
Nicola Sturgeon said Mr Rajoy's comments were not surprising.
She has been in Brussels for a series of talks with senior EU officials, after vowing to protect Scotland's interests in Europe following last week's Brexit vote.
She said she had received a "sympathetic response" but stressed she did not underestimate the challenges.
Any individual arrangement between Scotland and the EU would have to have the unanimous support of all EU member countries.
Mr Rajoy sounds serious about standing in the way of that. He said he wanted to be "very clear Scotland does not have the competence to negotiate with the European Union".
He added: "Spain opposes any negotiation by anyone other than the government of United Kingdom.
"I am extremely against it, the treaties are extremely against it and I believe everyone is extremely against it. If the United Kingdom leaves... Scotland leaves."
The French president seemed to concur. Francois Hollande said: "The negotiations will be conducted with the United Kingdom, not with a part of the United Kingdom".
Scotland’s Brexit referendum result was 62% in favour of staying and 28% wishing to leave.
Ms Sturgeon met European Commission president Jean-Claude Juncker in Brussels and EU Parliament president Martin Schulz. However, the European Council president Donald Tusk said a meeting would not be appropriate at this time.
