Moses Defending the Daughters of Jethro by Rosso Fiorentino is an oil on canvas painting dated to around 1523. The work illustrates a biblical episode in which Moses intervenes to protect the daughters of Jethro from shepherds at a well. This painting belongs to the early period of Rosso Fiorentino's career, during the height of the Italian Renaissance and the emergence of the Mannerist style. The composition is dominated by muscular, twisting male figures in the foreground, engaged in a violent struggle.

Moses, identifiable by his dynamic posture and raised arm, is depicted in the act of defending the women, who are shown to the right, one of them gesturing in alarm. The scene is set outdoors, with a backdrop of classical architecture and a blue sky. The artist employs vivid colors and strong contrasts of light and shadow, emphasizing the drama of the moment and the physicality of the figures. This painting is recognized as an important example of early Mannerism, reflecting Rosso Fiorentino's distinctive approach to anatomy and composition.

It has been widely studied for its innovative style and its influence on later sixteenth-century painting.