God's Little Acre presents two primary narratives. One storyline follows Ty Ty Walden, who is on a relentless quest for gold he believes is hidden on his land. The other focuses on his son-in-law, Will Thompson, who is determined to reopen a cotton mill that has been closed by the owners due to a workers' strike. Both characters are driven by their ambitions and are willing to sacrifice everything, including their families and their own lives, to achieve their dreams. Ty Ty's pursuit is particularly irrational. Despite not being wealthy, he owns land that could be used for farming. Instead, he neglects it, choosing to dig massive holes with his two sons, transforming his farm into a scene of destruction. Ty Ty represents, in an exaggerated form, the Southern farmer who has wasted his land through poor decisions, delusion, and pride, ultimately losing what little success he had. In contrast, Will Thompson is portrayed as a more heroic character. As a leader of the strike that has shuttered the mills in nearby Scottsville, Will is motivated by more than just personal pride. He feels a responsibility to his fellow workers, who depend on him to confront the oppressive mill owners. Through Will's story, Caldwell expands his exploration of social awareness, transitioning from the rural setting to the urban environment and from farmer to worker.

The book intricately weaves a prominent sexual theme throughout its storyline. Both Ty Ty and Will are driven by their physical desires. Ty Ty approaches his sexual longings with a philosophical mindset. In his later years, he finds real joy in simply admiring women, especially his daughter-in-law, Griselda. He believes that God played a "mean trick" by giving humans both a moral compass and strong primal instincts. Ty Ty expresses, "A man can't live, feeling himself from the inside, and listening to what the preachers say . . . He can live like we are made to live, and feel himself on the inside, or he can live like the preachers say, and be dead on the inside." He argues that the "natural" way to live is to acknowledge and embrace the sexual dimension of human nature. As a result, when he discovers that his daughter Darling Jill is sexually active, he celebrates her growth.

Will Thompson, in contrast, embodies a nearly messianic form of sexuality. On the eve of leading the strikers to the mill to "turn the power on"—an act that will lead to his downfall—three women care for him: his wife Rosamond, her sister Darling Jill, and their sister-in-law Griselda. Griselda, being the most physically and sexually commanding of the three, is expected to make love with Will before his looming death. This encounter is both intense and Dionysian, yet it also holds a ritualistic essence that Caldwell captures brilliantly. For Caldwell, as well as the characters in his narrative, the act of sex signifies an affirmation of life. Although it can be misused, it also connects humanity to its inherent divinity: the god of creation and vitality.