The Florida pinesnake is commonly associated with the longleaf pine ecosystem that once covered a large portion of the coastal plain in the southeastern United States. They occupy dry, upland areas with well-drained, sandy soils, characterized by pine-dominated or pine-oak woodland. Pinesnakes can also be found in pine flatwoods, oak scrub, dry oak forests, old fields and other ruderal habitats, particularly if southeastern pocket gophers are present. Florida pinesnakes exhibit a strong preference for pine forests, with open-canopy and frequent fire.

Florida pinesnakes are non-venomous, powerful constrictors that consume a variety of prey. Small mammals, including southeastern pocket gophers, as well as rabbits, and other rodents, make up a large portion of the pinesnake’s diet, but they are also known to eat ground-dwelling birds, bird eggs and lizards. Pinesnakes are active foragers that hunt, both above and below ground. 

Florida pinesnakes are large, heavy-bodied snakes with adult specimens typically ranging from 122 to 168 centimeters (4.5 to 5.0 feet) with a maximum total length of 229 centimeters (7.5 feet).

The Florida pinesnake displays a variety of morphological scale patterns and coloration, but the dorsum is typically gray, brown or rusty brown on a light gray or tan to cream background. The contrasting of colors tends to be less pronounced near the head and the contrast between the pattern and background color becomes more robust approaching the tail. The ventral side is white to yellow with moderate to little spotting as Tuberville and Mason noted in 2008. Coloration can vary regionally, with some being very pale or dark, and young Florida pinesnakes have very distinguished dark patches in their scale pattern, as noted by G.J. Miller and others in 2016. Overall, as Conant and Collins noted in 1998, the markings of the Florida pinesnake subspecies tend to not be as sharply defined as the northern pinesnake.

Pinesnakes are known for their loud hissing defensive behavior. When threatened, a Florida pinesnake may coil up, inflate its body, hiss loudly and create a rattling noise by rapidly shaking their tail in dry leaves. To produce the loud hissing sound, exhaled air is passed over a flap of tissue that stretches across the air opening in the mouth, which will be partially open. This visual and audible display partnered with their coloration can cause a Florida pinesnake to be misidentified as an eastern diamondback rattlesnake.

The Florida pinesnake is diurnal and highly fossorial, spending much of their time underground. Pinesnakes hibernate during the winter and may estivate in the summer depending on conditions. 

Courtship and mating occurs April through May, with female pinesnakes laying eggs May through July in underground burrows, as C. H. Ernst and E.M. Ernst noted in 2003. It is possible that timing of courtship and mating can vary based on latitude, but this is not currently documented in the literature. Florida pinesnakes excavate cavities for nest and sometimes use burrows of other animals, such as southeastern pocket gophers (Geomys pinetis), gopher tortoises (Gopherus polyphemus), and nine-banded armadillos (Dasypus novemcinctus).