In order to delay the occurrence of browning of fresh-cut Agaricus bisporus under cold storage, effects of filling nitrogen in different vacuum (0, -20, -40, -60, -80, -98kPa) combined with heat treatment on color, enzyme activities of peroxidase, polyphenol oxidase and phenylalanine ammonia-lyase, and total phenolic content of fresh-cut Agaricus bisporus were evaluated. The results showed that filling nitrogen in vacuum combined with heat treatment slowed down the decrease rate of L* value (brightness) of fresh-cut Agaricus bisporus, inhibited its color development to red and yellow, and finally delayed browning of fresh-cut Agaricus bisporus. In comparison with control (0kPa) and other filling nitrogen in vacuum combined with heat treatment, both -40kPa and -80kPa of filling nitrogen in vacuum combined with heat treatment (hot-water temperature 40℃ and heat treatment time 5min) kept the POD and PPO activities at a lower level, and maintained PAL activity and total phenolic content at a higher level of fresh-cut Agaricus bisporus, which also delayed the senescence and improved its resistance of fresh-cut Agaricus bisporus.