The Altarpiece of Our Lady of the Rosary was commissioned in 1593 by Sister María de la Resurrección, a professed nun at the monastery. Like the previously mentioned altarpieces, it follows a triumphal-arch architectural model, and its authorship is attributed to Miguel Adán.

The structure of the altarpiece is divided into two main sections: the lower body, consisting of the arch, the sculptural figures in the round, and a panel painting; and the upper body, or attic.

In the lower section, the central figure is Our Lady of the Rosary, sculpted in the round and placed within a rectangular niche atop a pedestal. She is flanked by statues of Saint Dominic and Saint Thomas Aquinas—the latter holding a small temple symbolizing the Church—both positioned in niches with curved tops. Surrounding this central sculptural group are the fifteen Mysteries of the Rosary, with the Evangelists and Church Fathers positioned on the pedestal and the most prominent Dominican and Franciscan saints placed along the arch’s curve.

Above the sculpture of Our Lady is a late fifteenth-century panel painting of Saint Mary of the Populo. On either side of this panel are two reliefs depicting the Tota Pulchra and the Immaculate Conception, while the spandrels of the arch feature Saint John the Baptist and Saint Dominic, as well as Saints Catherine and Barbara. Additional reliefs depict the sorrowful Mysteries on the right, the glorious Mysteries in the attic, and the joyful Mysteries on the left.

The attic is divided into five sections, each topped with alternating triangular and curved pediments. The central section, the tallest, represents the Coronation of Our Lady by the Holy Trinity—the fifth glorious Mystery of the Rosary—and is crowned with a decorative coat of arms. Adjacent reliefs depict angels, while the extreme sections illustrate the fifth joyful Mystery on the left and the sorrowful Mystery on the right.

The iconography of the altarpiece is entirely Dominican in character. The Virgin of the Rosary is surrounded by the fifteen Mysteries—joyful, sorrowful, and glorious—intended to guide the faithful in meditation as they prayed. The work demonstrates pronounced classicism, meticulous attention to detail, and a careful use of polychromy, enhancing the realism of the figures.

Some of the depicted scenes include: on the base, Eve; Saints John and Luke; Saints Gregory and Jerome; Saint Dominic receiving the pastoral staff and the Epistles of Saint Peter and Saint Paul; Saints Augustine and Ambrose; Saints Matthew and Mark; and Adam. On the arch’s curve are the Birth of the Virgin Mary; Saints Pius V, Diego de Alcalá, and Vincent Ferrer; Saints Francis and Peter Martyr; Saints Anthony, Telmo, and Bonaventure; and the Presentation of the Blessed Virgin Mary. In the attic, two angels are depicted. Finally, the altarpiece completes its iconographic program with the joyful, glorious, and sorrowful Mysteries of the Rosary.