Born in South Carolina to parents who had been slaves and having to work in fields at age five, she took an early interest in her own education. With the help of benefactors, Bethune attended college hoping to become a missionary in Africa. When that did not materialize, she started a school for African-American girls in Daytona Beach. From six students it grew and merged with an institute for African-American boys and eventually became the Bethune-Cookman School. Its quality far surpassed the standards of education for African-American students, and rivaled those of schools for white students. Bethune worked tirelessly to ensure funding for the school, and used it as a showcase for tourists and donors, to exhibit what educated African-Americans could do. She was president of the college from 1923 to 1942 and 1946 to 1947, one of the few women in the world who served as a college president at that time.
Bethune was also active in women's clubs, and her leadership in them allowed her to become nationally prominent. She worked for the election of Franklin D. Roosevelt in 1932, and became a member of Roosevelt's Black Cabinet, sharing the concerns of black people with the Roosevelt administration while spreading Roosevelt's message to blacks, who had been traditionally Republican voters. Upon her death, columnist Louis E. Martin said, "She gave out faith and hope as if they were pills and she some sort of doctor." Her home in Daytona Beach is a National Historic Landmark, her house in Washington, D.C. in Logan Circle is preserved by the National Park Service as a National Historic Site, and a sculpture of her is located in Lincoln Park in Washington, D.C.
Most of her siblings were born into slavery. Her mother worked for her former owner, and her father farmed cotton near a large house they called "The Homestead."
After demonstrating a desire to read and write, McLeod attended Mayesville's one-room schoolhouse, Trinity Mission School that was run by the Presbyterian Board of Missions of Freedmen. Her teacher, Emma Jane Wilson, became a significant mentor in her life. Wilson had attended Scotia Seminary (now Barber-Scotia College), so arranged for McLeod to attend the same school on a scholarship, which she did from 1888-1893. The following year, she attended Dwight L. Moody's Institute for Home and Foreign Missions in Chicago (now the Moody Bible Institute), hoping to become a missionary in Africa. However, she was told that she would not be able to go because black missionaries were not needed, so she instead planned to teach.
She married Albertus Bethune in 1898 and they subsequently lived in Savannah, Georgia, for a year while she did some social work. She was persuaded by a visiting Presbyterian minister named Coyden Harold Uggams (grandfather of entertainer Leslie Uggams) to relocate to Palatka, Florida, to run a mission school.She did so in 1899 and began an outreach to prisoners and ran the mission school. Albertus left the family in 1907 and did not seek a divorce, but relocated to South Carolina. He died in 1918.
Bethune was influenced deeply by Delaney and adopted many of her educational philosophies seeking to improve the conditions of black people by educating primarily women: "I believe that the greatest hope for the development of my race lies in training our women thoroughly and practically." After one year at Haines, Bethune was transferred to the Kindell Institute in Sumter, South Carolina, where she had met her husband.