This small panel by Lotto can be dated to early in his career because of stylistic similarities to Allegory of Virtue and Vice, also in the National Gallery, signed by Lotto and dated 1505. That work originally functioned as the cover of a portrait, now in Naples, of Bernardo de' Rossi, bishop of Treviso. The Allegory of Chastity probably served a similar purpose. Covers not only protected the painting underneath but also allowed the artist to expand on particular facets of the patron's personality or concerns.
The subject of the panel has been the cause for much discussion. A variety of interpretations have been suggested, including the classical, Plutus and the Nymph Rhodos, and the literary, Petrarch's poetic vision of Laura. A cupid showers her with flower petals, while a female satyr gazes around a tree at a male satyr, who is more interested in his jug of wine. The composition is divided horizontally in two parts, the maiden and cupid seen on one level, the satyrs on a lower, baser level. It can be read as a choice between virtue and pleasure, virtus and voluptas, the chaste maiden in repose, or the satyr couple, symbolic in the Renaissance of lust and even evil. It was considered quite appropriate to associate such moralizing themes with portraits.