Background: The rate of respiration of each agricultural commodity varies significantly from commodity to commodity and cultivar to cultivar. To build a controlled, modified or cold storage system and determine shelf life, it is necessary to quantify the rate of respiration and the heat generated as a result of respiration. Methods: A scientific study was carried out to determine the effect of different storage conditions on the rate of respiration and heat of respiration of mango fruits (cv. Alphonso). An air-tight multi-chamber temperature and RH control system was developed to experiment with various temperatures. Mango fruits were stored at 10, 15, 20, 25°C and ambient temperature in the developed air-tight multi-chamber system. The rate of respiration and heat of respiration was determined for different storage conditions. Result: At the beginning maximum rate of respiration, 63.22, 72.56, 81.13, 86.33, 101.55 ml CO2/kg/h and heat of respiration, 7096.53, 8322.05, 9119.29, 9589.19, 10547.55 kcal/metric ton/day was observed at 10, 15, 20, 25°C and ambient temperature respectively. It was found that under steady-state storage conditions the rate of respiration and heat of respiration was increased with an increase in temperature whereas decreased with time for all storage conditions.