The nutrients found in Plantain are necessary for the body's metabolism. However, if the fruits are infected with fungi, they can be tampered with. This can lead to a reduction in the nutrients' quality and quantity. Fruit rot of Plantain is a post-harvest fungi that can affect the quality and quantity of the fruit. The objective of the study was to determine the fungi associated with fruit rot of plantain and its effects on the nutritional value of plantain fruits. Three samples of healthy plantain fruits were taken from Eke-Aka market in Anambra state and were transported to Alpha research laboratory Awka, Anambra state in a sterile polythene bag for isolation. Proximate analysis was carried out on both samples to determine the moisture content, crude fat, crude fibre, crude protein, ash content and carbohydrate content determination. The fungi associated with the spoilage of Plantain fruits in Eke Awka, Anambra state were studied using the standard procedure. Potato dextrose agar (PDA) and Sabouraud dextrose agar (SDA) were the culture media used. 39g of PDA and 69g of SDA were poured into one litre of water in the conical flask and autoclaved at 121ºC at 15psi for 20 minutes. Fungi identified were: Aspergillus sp, Fusarium sp and Penicillium sp. The result showed that Aspergillus sp (52.50%) had the highest frequency of occurrence from the fungi isolated while Penicillium sp. (20.00%) had the least occurrence. The result of nutritional analysis showed that the healthy plantain pulp had high moisture content (27%) and low fibre (6.50%) while the spoilt plantain pulp have high moisture content (35.20%) and low protein content (1.30%). From the study, the activity of these microorganisms affect the nutritional value of the plantain fruit.