The change, arriving this week, was sanctioned after a revision of infection rates in mainland Spain and the Spanish islands.
The move paves the way for Scots to go on vacation to Spain and Spaniards traveling to Scotland.
But he warned that Covid-19 was “still active and still deadly” and added that further changes would only be made when it was “safe to do so”.
People traveling to the UK from abroad are kept in quarantine for two weeks on arrival or face a fine.
However, a list of countries in Europe and the world have been exempted from this, creating “air bridges” to allow people to travel more freely.
The Scottish government – which has control over public health measures north of the border – initially did not include Spain in its list of countries free of self-isolation requirements, citing concerns about the prevalence of coronavirus there.
A “thorough review” has now been conducted and means that Spain and its islands will be added to the list later in the week.
Yousaf said: “Following this latest review and after carefully evaluating the potential public health impact of changing the national quarantine exemptions, we are able to remove this requirement for those traveling from Spain. We continue to keep other countries under control and where there is clear evidence that it is safe to do so, we will add other countries
“Clearly the virus has not gone away. It is still active and still deadly. With the quarantine restrictions lifted by these countries this means that we need to be even more careful about what we are doing.
“Anyone traveling should always follow public health advice, including putting on facial clothing, avoiding crowded places, washing hands and surfaces, staying two meters away and self-isolating if symptoms occur and registering for a test immediately.”
Where can I go without quarantining on my return?
As of Friday, travelers are exempt from quarantine rules when arriving in Scotland from:
Andorra, Antigua and Barbuda, Aruba, Australia, Austria, Bahamas, Barbados, Belgium, Bonaire, Sint Eustatius and Saba, Croatia, Curaçao, Cyprus, Czech Republic, Denmark, Dominica, Faroe Islands, Fiji, Finland, France, French Polynesia, Germany, Greece, Greenland, Grenada, Guadeloupe, Hong Kong, Hungary, Iceland, Italy, Jamaica, Japan, Liechtenstein, Lithuania, Luxembourg, Macao, Malta, Mauritius, Monaco, Netherlands, New Caledonia, New Zealand, Norway, Poland, Reunion, San Marino, Seychelles, South Korea, Spain, St Barthélemy, St Kitts and Nevis, St Lucia, St Pierre and Miquelon, Switzerland, Taiwan, Trinidad and Tobago, Turkey, Vatican City, Vietnam.