Rural home garden is an important traditional source of medicinal plants for daily curative uses throughout Bangladesh. Such knowledge is continuing from generation to generation without documentation. An ethnobotanical investigation was conducted through focus group discussions and households’ survey accompanied by field observation to document the indigenous knowledge of herbal medicines being used by the rural communities of Comilla district in Bangladesh. A total of 45 ethnomedicinal plant species belonging to 34 families were found, where trees (37.78%) were the most commonly utilized growth form. Plant resources are used to treat 31 different individual ailments ranging from simple cuts to heart disease. Plants are mainly used to treat dysentery (12 species), cold ailments, cough, and fever (6 species each). For curing ailments, the use of the above ground plant parts was higher (86.44%); particularly fruits (37.29%) and leaves (30.51%) were the most commonly used plant parts. More than half of the medicinal plants are indigenous (71.11%), being edible fruit bearer (48.89%), plants parts suitable for animals and birds (57.78%), and natural regeneration present (64.44%) indicated that medicinal plants play a vital role in biodiversity conservation in the study area.