Scottish Government agencies in charge of creating jobs and attracting tourists have become the latest to quit business lobbying organisation the CBI after it formally backed a "no" vote in the referendum.
The government in Edinburgh said it would be "clearly inappropriate" for Scottish Enterprise and VisitScotland to carry on their membership after the body registered with the Electoral Commission to campaign against independence.
The two agencies follow broadcaster STV, the Balhousie Care Group and energy company Aquamarine Power in resigning from the CBI.
A Scottish Government spokesman said: "The CBI has registered as a campaign organisation for a 'no' vote in the referendum.
"In these circumstances, it is clearly inappropriate for government agencies to remain in membership of CBI.
"We would expect any other member to follow Scottish Enterprise and resign with immediate effect."
The government also released statements from the two agencies.
Scottish Enterprise said: "In light of CBI Scotland's political decision to register with the Electoral Commission, Scottish Enterprise have had no choice but to immediately resign from the organisation"
A VisitScotland spokesman said: "In the light of the decision of the CBI to register with the Electoral Commission, VisitScotland has decided it is appropriate to withdraw from the organisation."
It emerged on Friday that CBI Scotland, which represents many businesses across the country, had registered with the Electoral Commission, meaning it can spend more than £10,000 on campaigning during the referendum period.
Registering as a campaigner also gives access to the electoral register and the right for representatives to attend postal vote opening sessions, polling stations and the counting of votes.
The confederation faced criticism that its position does not accurately reflect its members' views, and it failed to consult them before formally registering to campaign for the union.
A CBI spokesman said that Scotland and the rest of the UK are "stronger together as part of the union" and its view reflects the "vast majority" of members.
CBI director Iain McMillan gave his views on the implications of independence in an appearance before Holyrood's Economy Committee earlier this month.
He said: "This would not be a land of milk and honey. It would be extremely difficult with many painful decisions to be taken."
Broadcaster STV revealed its decision to quit in a statement on its website.
"STV is a public service broadcaster with a duty of impartiality and as such we have no corporate or editorial position on the independence referendum in September," it said.
"In light of CBI Scotland's decision to register with the Electoral Commission we have no choice but to resign our membership of CBI Scotland forthwith."
Tony Banks, chairman of the Balhousie Care Group, quickly responded to the CBI's position on Friday and said his business will now leave the confederation.
Mr Banks, who is also chairman of the pro-independence group Business for Scotland, said: ''It is abundantly clear that the CBI is not representing its members' views honestly. I am therefore writing to the director-general, John Cridland, today, withdrawing my company membership.''
Martin McAdam, chief executive officer of Aquamarine Power, said there had been no consultation before the CBI decision.
Electrical contractors union Select, which represents 1,250 companies with 18,500 employees, has also demanded the CBI reviews its decision.
While there has been no threat to leave the CBI, the union said the Better Together affiliation does not reflect its views.