- Lena Grove – a young pregnant woman from Alabama who has traveled to Jefferson while looking for Lucas Burch, the father of her unborn child.
- Byron Bunch – a bachelor who works at the planing mill in Jefferson, who meets and falls in love with Lena when she arrives in town. She has been told that a man named Bunch works at the mill and assumes it is Lucas, because the name sounds similar.
- Gail Hightower – the former minister of Jefferson, forced to retire after his wife was discovered to be having an affair in Memphis and committed suicide. He is a friend and mentor to Byron.
- Lucas Burch/Joe Brown – the young man who fathered Lena's child in Alabama and ran away when she told him she was pregnant. He has been living in Jefferson with Joe Christmas in a cabin on Joanna Burden's property under the name Joe Brown and working with Christmas and Byron at the planing mill. He is also a bootlegger.
- Joe Christmas – a man who came to Jefferson three years prior to the events in the novel. He lives in a cabin on the property of Joanna Burden and has a secret sexual relationship with her. Although he has light skin and is an orphan with no knowledge of his family background, he believes that one of his parents are of African-American ancestry, and this secret has caused him to be a habitual wanderer. He is employed at the planing mill until he begins to make a profit as a bootlegger.
- Joanna Burden – the sole survivor in Jefferson of a family of abolitionists from New England who came to Jefferson after the Civil War. She is unmarried, lives alone in a manor house outside of Jefferson, and is secretly engaged in a sexual relationship with Joe Christmas. She is murdered, presumably by Christmas, at the start of the novel, and her house is burned down.
- Eupheus "Doc" Hines – the grandfather of Joe Christmas. He hates Christmas and gives him away to an orphanage when he is born, staying on as a janitor there in order to monitor the boy. Later, when he hears that Christmas is being held on suspicion of murdering Joanna Burden, he travels to Jefferson with his wife and begins to incite a lynch mob to kill Christmas.
- Mrs. Hines – the grandmother of Joe Christmas. She has never seen Christmas after the night of his birth and travels to Jefferson to ensure that her husband does not successfully have him lynched, because she wants to see him again once more before he is tried for murder.
- Milly Hines – the teenage mother of Joe Christmas. She conceives after a tryst with a member of a traveling circus, whom she claims is Mexican. She dies in childbirth after Eupheus Hines refuses to call a doctor for her.
- Mr. McEachern – the adoptive father of Joe Christmas. He is a devout Presbyterian and tries to instill religion in the young orphan he has adopted. He disapproves of Christmas's growing disobedience and is killed by his adopted son when the boy is 18.
- Mrs. McEachern – the adoptive mother of Joe Christmas. She tries to protect Christmas, though he hates her and pulls away from her attempts to be kind to him.
- The dietitian – a woman who worked at the orphanage where Joe Christmas was raised. After he accidentally sees her with a man in her room, she tries unsuccessfully to have him transferred to an all-black orphanage.
- Mr. Armstid – a man who picks up Lena on her way to Jefferson, lets her spend the night at his house, and then gives her a ride to the city on his wagon.
- Mrs. Armstid – Armstid's wife, who gives Lena money in spite of her disdain for the young woman.
- Bobbie – a waitress at a restaurant in Memphis whom the adolescent Joe Christmas falls in love with and proposes to on the night that he kills his father at a local dance. She scorns him and leaves him.
- Gavin Stevens – an educated man and district attorney who lives in Jefferson and offers commentary on some of the events at the end of the novel.
- Percy Grimm – the captain of the State National Guard who kills Joe Christmas and castrates him.
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