Currently, RoiTools supports union and intersection methods directly but not subtraction.
Subtraction has to be done using combineROIs and CombineOp methods, while not difficult, is incoherent as union and intersection is directly available as mentioned above.
I would like to suggest having a RoiTools.subtract(roi1, roi2) method added alongside RoiTools.union(rois) and RoiTools.intersection(rois).