The Florida Railroad was the state's first cross-peninsular railroad. David L. Yulee, a Florida resident and United States Senator, incorporated the Florida Railroad Company in 1853. Construction of the line began at Fernandina Beach in 1856 and was completed to Cedar Key in 1861, covering a distance of 155 miles. The railroad passed through the present-day communities of Baldwin, Starke, Waldo, Gainesville, Archer, and Bronson. Railroad stations in Nassau County included Fernandina Beach, O'Neil, Lofton, Yulee, Whittsville, Italia, Callahan, Crawford, Dahoma, Inglehame, and Bryceville. The railroad roughly parallels present-day State Road 200 from Fernandina Beach to Callahan and present-day US 301 from Callahan south to the Nassau County line.
Intensive manual labor was required to construct the Florida Railroad. The company recruited slaves and free laborers to fell trees, drive spikes, and lay crossties and rails. The railroad used a broad gauge of five feet between rails with heavy rail generally of sixty pounds per linear yard. Crossties were made of yellow pine and were eight feet long, seven inches wide, and seven inches high. Wooden bridges were constructed across Nassau County's many creeks and rivers, including Kingsley Creek, Boggy River, Lofton Creek, Mills Swamp, and Plummer Swamp.
Completed in 1861, the railroad allowed ships from the ports of the eastern United States to avoid the lengthy and often dangerous passage through the Keys to reach the markets of the Gulf coast. Ship cargoes were unloaded in Fernandina Beach for transportation across the state via the railroad to Cedar Key. From there, goods were reloaded and carried to New Orleans, Mobile, and elsewhere. The railroad also benefited from the agricultural enterprises of the state's interior, which included sugar, cotton, tobacco, cattle, and fruits and vegetables. Naval stores and timbering, both of which were emerging industries at the time, also benefited from the railroad.
As Florida's roadways modernized in the twentieth century, the importance of the railroad declined. Tracks were removed in parts of Nassau, Alachua, and Levy Counties beginning in the 1930's. The railroad is still active in Nassau County from Fernandina Beach to Yulee and from Callahan south to the county line. Remains of the abandoned railroad can still be found along the 15-mile segment between Yulee and Callahan.