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There could be several reasons why audio is not playing in release mode on Flutter:

1. Missing Assets: Check if the audio files are properly included in the assets section of your pubspec.yaml file. Make sure that the files have the correct path and file name.

2. Incorrect Code: There might be an error in your code where the audio is not being played properly. Verify that the code is correct and has been implemented correctly.

3. Audio Format: Some audio formats might not be supported on some devices, make sure that you are using a supported format for the target devices.

4. Permission Issues: Ensure that you have granted the required permissions to access the device's audio.

5. Third-Party Dependencies: There might be a dependency conflict that is causing the audio to not play in release mode. Ensure that you are using the correct version of the dependencies required for playing audio.

6. Testing Environment: Sometimes, the issue might not be related to code or implementation but might be related to the device or the testing environment. Try testing on multiple devices and platforms to identify the issue.