Vicente del Bosque's men swept to a 4-0 victory over Italy in the 2012 final held in Kiev, to back up their 2008 success under previous boss Luis Aragones.
Spain are currently enjoying one of the most dominant eras in history, having also collected the FIFA World Cup in South Africa in 2010.
Ukraine - co-hosts of the 2012 tournament with Poland - will hope to provide a stern test of the holders' credentials, while Luxembourg, the Macedonia and Belarus complete Group C.
Gibraltar are appearing in their first UEFA competition and were originally drawn to appear in Group C.
However, the island nation will instead play in Group D, as a decision had already been taken to keep them apart from Spain for political reasons.
That means Gibraltar face a mouth-watering encounter with another of the pre-tournament favourites in Germany, three-time winners of the competition.
Near neighbours Poland, along with the Republic of Ireland, appear likely to provide the biggest threat in Germany's group, while Scotland and Georgia also await Joachim Low's side.
Italy were runners-up in 2012 and face difficult Group H encounters with Croatia, Norway and Bulgaria in their attempt to go one better this time around.
Armenia were also due to play in that pool, but were moved to Group I as they cannot play in the same group as Azerbaijan due to political reasons.
In Group E, top seeds England were drawn to meet Switzerland, meaning Roy Hodgson will face a reunion with the nation he enjoyed a successful spell in charge of between 1992 and 1995.
The Netherlands, winners in 1988, were the first team to be drawn out and face challenges from the Czech Republic and Turkey in Group A.
Portugal feature in Group I along with Denmark, Serbia, Armenia and Albania.
The draw saw 53 nations split into six pots and subsequently drawn into eight groups of six and one of five. France will be partnered with the five-team group, enabling the 2016 tournament hosts to play friendlies against these countries on their 'spare' dates
The top two sides from each group will qualify automatically for the finals, together with the third-placed team with the best record against the nations first, second, fourth and fifth in their pool.
Play-offs will then be used to determine which of the eight remaining third-placed sides will make it to France.
The qualifiers will run from September 2014 to October 2015, with matches taking place between Thursday and Tuesday as part of UEFA's new 'Week of Football' format, which replaces the previous model where games were played on a Friday and Tuesday.
Euro 2016 qualifying draw in full:
Group A: Netherlands, Kazakhstan, Iceland, Latvia, Turkey, Czech Republic
Group B: Bosnia-Herzegovina, Andorra, Cyprus, Wales, Israel, Belgium
Group C: Spain, Luxembourg, Macedonia, Belarus, Slovakia, Ukraine
Group D: Germany, Gibraltar, Georgia, Scotland, Poland, Republic of Ireland
Group E: England, San Marino, Lithuania, Estonia, Slovenia, Switzerland
Group F: Greece, Faroe Islands, Northern Ireland, Finland, Romania, Hungary
Group G: Russia, Liechtenstein, Moldova, Montenegro, Austria, Sweden
Group H: Italy, Malta, Azerbaijan, Bulgaria, Norway, Croatia
Group I: Portugal, Albania, Armenia, Serbia, Denmark, France
