After the consecration of the Church of the Nativity of Christ, the venerable Seraphim addressed M.V. Manturov with a remark: "We, father, have built a church in honor of the Nativity of the Savior, but we do not have a church in honor of the Virgin! The Queen of Heaven has become angry with me, poor Seraphim, and said: 'You honored My Son, but forgot about Me!' Therefore, I thought, father, should we not build another church under the temple? Try, father, and we will have two churches." The venerable one gave a thread for measurement, saying that if the tunnel under the Church of the Nativity of Christ coincided with the measure, then that is where we should build the church in honor of the Nativity of the Mother of God. The tunnel turned out to be accurate, but the ceiling was low and required supporting pillars, and the church turned out to be small. Manturov sought advice from Seraphim, who, filled with joy, exclaimed: "Four pillars, that means the relics will rest here! This will be a tomb for the relics, father!" In 2000, a tomb for the relics of the holy women of Diveevo, Alexandra, Martha, and Elena, was indeed arranged here.

The construction of the church in honor of the Nativity of the Virgin was completed in the summer of 1830. On September 8, the temple was consecrated by Archimandrite Joachim from the Annunciation Monastery, who came to Diveevo despite the cholera epidemic. The iconostasis in the church was single-tiered, resembling that of the Nativity of Christ, but painted green, with the Crucifixion on the Royal Doors. The temple was warm, and the floor in the lower church was decorated with multi-colored ceramic tiles, forming a pattern reminiscent of a carpet.

After the monastery was closed, the church was used as a storage facility. The re-consecration of the lower church took place on October 21, 1992, and in 1993, the first monastic tonsure after the reopening of the monastery was held.

By the command of the venerable Seraphim, a lamp burns constantly by the icon of the Nativity of the Virgin, and the sisters read the unceasing Psalter. In 2008, chapels in honor of Archangel Michael and the Royal Martyrs were added to the church. The altars of the lower chapels contain the graves of Mikhail Vasilyevich Manturov and the spiritual father of the Diveevo sisters, Archpriest Vasily Sadovsky.

The church is decorated with frescoes by Moscow artist Konstantin Okhotin, dedicated to the earthly life of the Most Holy Theotokos and the history of the Diveevo monastery. The chapels are painted by artists from the Nizhny Novgorod workshop "Ark."