The present investigation was carried out to characterize the pathogen responsible for Choanephora fruit rot disease in bitter gourd along with find out appropriate control measure using biological resources. The pathogenic fungal strain was isolated from the disease infected fruit part of bitter gourd. It was subjected to four different types of growth media named potato dextrose agar (PDA), Czapeck Dox agar (CDA), nutrient agar (NA) and Sabouraud dextrose agar (SDA) were used to note down the differences regarding to various characteristics appeared due to dissimilation of culture medium. Thereafter, the growth environment and medium contents were changed to evaluate the effect of temperature, pH, carbohydrate, sugar concentration, salt, organic acid etc. on the mycelial growth of the concerned fungus. In the experiment, these parameters significantly influenced the growth pattern of the fungus. For proper identification of the fungus, advanced molecular technique was used. The PCR product of the internal transcribed spacer (ITS) region of the fungi showed nearly 650 bp size of clear band followed by sequencing of the PCR product of the fungus. The sequenced data showed 99% similarities with the original sequence of Choanephora cucurbitarum. In search of suitable biocontrol measure, the antifungal activities of the seven plant extracts namely Hibiscus rosa-sinensis, Ficus racemosa, Moringa oleifera, Psidium guajava, Azadirachta indica, Cassia alata and Coccinia grandis were assayed where Azadirachta indica (42.12%) and Moringa oleifera (35.71%) had promising inhibitory effects against the isolated fungus. The outcome of the experiment would be benevolent for the detection and development of eco-friendly control approach of the detrimental disease of the bitter gourd.