TL;DR - this probably won't help but it's all I've got for now.
Blind leading the blind here, but I found this way old thread with a FreeDOS initdisk error: https://www.ultimatebootcd.com/forums/viewtopic.php?t=1545
For them it was supposedly a memory management issue. They also had opcode errors displayed on screen... this is a total shot in the dark, but the FreeDOS download page has a note on getting past opcode errors in Virtual Box (I know you're on real hardware):
>If you get a scrolling list of “Invalid Opcode” errors when you boot the FreeDOS 1.2 installation CD-ROM on VirtualBox, this is a bug in VirtualBox since we released FreeDOS 1.2.
>To fix: When you boot the FreeDOS install CD-ROM, at the first “Welcome to FreeDOS 1.2” boot screen, press the Tab key to edit the boot options. Add raw to the end of the command line, then press the Enter key to boot.
Example: https://www.freedos.org/download/screenshot-raw.png
If I've got to just guess, perhaps the hardware (particularly RAM) is more advanced than FreeDOS can handle without extra configuration? Maybe see if you can limit how much RAM FreeDOS will use, try booting without extended memory?
Yes, but only through UHCI and some EHCI USB slots, so on a relatively older computer (e.g. prior to 2009) there's a high chance it will work, while on a 2018 motherboard it's practically 100% sure it won't. Albeit ext2 and NTFS can be read, it's obviously best to format it FAT16/32. There was a quick write up on the topic I had shared for someone else a while ago, you can get it here