Barbora Strycova takes on Garbine Muguruza in Saturday's opening rubber of the first-round Fed Cup tie as the defending champions host Spain.
World number 17 Strycova will face 7th-ranked Muguruza as the Czechs start their quest for a fourth straight title in the eastern Czech steel hub of Ostrava.
Third-ranked Karolina Pliskova of the Czech Republic will then play against 70th-ranked Lara Arruabarrena.
Pliskova and Strycova led the Czechs to their third Fed Cup title in a row — and fifth in six years — last November when the team beat France in Strasbourg 3-2.
Spain have won five titles so far, the last one in 1998.
"I think we should succeed if the girls play their standard game," Czech captain Petr Pala said after the draw.
"Barbora will have to break her opponent's rhythm in some way, and to change hers so (Muguruza) wouldn't know what will come next."
In Sunday's reverse singles, Pliskova will face Muguruza in a clash of the number-one players and Strycova is then due to take on Arruabarrena.
Czechs Lucie Safarova and Katerina Siniakova were drawn for the doubles rubber against Sara Sorribes Tormo and Maria Jose Martinez Sanchez.
The Czechs miss 12th-ranked two-time Wimbledon champion Petra Kvitova, who is recovering from a hand injury she suffered when fighting a knife-wielding burglar last December.
Spain miss world number 14 Carla Suarez Navarro over a right shoulder injury.