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We have an XP Pro host, DOS guest, and the latest VirualBox.

Is it possible for the DOS guest to be installed on a physical partition? For example, if the XP host has a 512 MB O: partition, could we install and run the DOS guest on this partition, instead of using a .vhd or .vdi file?

Yes you could but you will have to remove the O: drive letter association or it will get corrupted.

I've been reading the manual and documentation, but I can't find any guides on how to set up a guest os to use a host os partition. The only hard drive setup options I've seen are to virtual hard drive files.

What steps should I take to allow the DOS guest to use a partition on the XP host?

Thanks.

Look for RAW disk access in the Manual.