The principal results in this paper are concerned with the description of domains of infinitesimal generators of strongly continuous groups of isometries in non-commutative operator spaces E(M, τ), which are induced by R-flows on M. In particular, we are concerned with the description of operator functions which leave the domain of such generators invariant in all symmetric operator spaces, associated with a semi-finite von Neumann algebra M and a separable function space E on (0, ∞). Furthermore, we apply our results to the study of operator functions for which [D, x] , where D is an unbounded self-adjoint operator. Our methods are partly based on the recently developed theory of double operator integrals in symmetric operator spaces and the theory of adjoint C 0 -semigroups.