We compute X-ray emission lines from thermal plasma in hot accretion flows. We show that line profiles are strong probes of the gas dynamics, and we present line-ratio diagnostics which are sensitive to the distribution of mass with temperature in the flow. We show how these can be used to constrain the run of density with radius, and the size of the hot region. We also present diagnostics which are primarily sensitive to the importance of recombination versus collisional ionization, and which could help discriminate ADAFs from photoionization-dominated accretion disk coronae. We apply our results to the Galactic center source Sagittarius A^* and to the nucleus of M87. We find that the brightest predicted lines are within the detection capability of current X-ray instruments.