The history of the parish dates back to the early 18th century. In 1710, Boone's Chapel was founded, and it is said to have been the place of Baptism of the late Baltimore Archbishop John Carroll, who in 1789 became the nation's first Catholic bishop. The chapel, which eventually led to the establishment of Most Holy Rosary Church, was used until about 1842. Since then, time and weather have deteriorated the chapel completely, and now a giant oak tree, with the circumference of 25 feet at the bottom, stands as a memorial of the historic chapel. A few markers and monuments still stand in the cemetery, as well as a few wooden crosses. Some have names, but many others remain unmarked.
It was at this time that a brick and frame church was built on Main Street in Upper Marlboro, known as St. Mary of the Assumption Church. Residents in the area attended St. Mary of the Assumption Parish in Upper Marlboro, until in 1858 when land was donated by Henry and Mary Mitcail for Most Holy Rosary Parish. According to the deed, the rural mission church was to be built within five years near the corner of Route 301 and Rosaryville Road. The cornerstone of the church was laid on a Sunday evening on June 8, 1858. The church was dedicated as a mission church 160 years ago, on Oct. 23, 1859. During the year after a violent storm demolished the church, Mass was held in the Knights of Columbus and Ladies Auxiliary Hall. In May of 1928, a new church was completed and dedicated. In 1966, Holy Rosary became a parish.
The parish's St. Joseph Center is located next to the rectory and is used for daily Mass on weekdays and a late morning Mass on Sunday. The main church is used for one morning Mass on Sunday (9 AM), and the vigil Mass on Saturday (5 PM). Most Holy Rosary Parish has been involved in the SHARE food distribution network for the past 16 years. The parish also supports the Upper Marlboro Food Bank with monthly food collections for the poor. The Adopt-A-Family program has been a popular program at the parish for the past 16 years. Members of the church provide meals for families in need during Christmas and Thanksgiving.
Father Isadore Dixon, recently retired has been the pastor of Most Holy Rosary Church for 18 years. As of July 1, 2007, Most Holy Rosary parish was placed in the capable hands of Parochial Administrator, Reverend Roger A. Soley. The other priests in the parish include: Paulist Father John Lynch, a retired professor of canon law at the Catholic University of America who has served at the parish for over 25 years, and Father John Smolko, a retired professor of theology at Catholic University who has served at the parish for 13 years. More recently, Father Smolko permanently retired his services at Holy Rosary Parish.
In 2008, Father Roger A. Soley was officially assigned as pastor of Holy Rosary Parish by Archbishop Donald Wuerl and remained there until July 10, 2019 when he was followed by Father Rodolfo (Rudy) Salinas.