<jats:title>Abstract</jats:title> <jats:p>Motivation: Reconstruction of gene-content evolutionary history is fundamental in studying the evolution of genomes and biological systems. To reconstruct plausible evolutionary history, rates of gene gain/loss should be estimated by considering the high level of heterogeneity: e.g. genome duplication and parasitization, respectively, result in high rates of gene gain and loss. Gene-content evolution reconstruction methods that consider this heterogeneity and that are both effective in estimating the rates of gene gain and loss and sufficiently efficient to analyze abundant genomic data had not been developed.</jats:p> <jats:p>Results: An effective and efficient method for reconstructing heterogeneous gene-content evolution was developed. This method comprises analytically integrable modeling of gene-content evolution, analytical formulation of expectation-maximization and efficient calculation of marginal likelihood using an inside-outside-like algorithm. Simulation tests on the scale of hundreds of genomes showed that both the gene gain/loss rates and evolutionary history were effectively estimated within a few days of computational time. Subsequently, this algorithm was applied to an actual data set of nearly 200 genomes to reconstruct the heterogeneous gene-content evolution across the three domains of life. The reconstructed history, which contained several features consistent with biological observations, showed that the trends of gene-content evolution were not only drastically different between prokaryotes and eukaryotes, but were highly variable within each form of life. The results suggest that heterogeneity should be considered in studies of the evolution of gene content, genomes and biological systems.</jats:p> <jats:p>Availability: An R script that implements the algorithm is available upon request.</jats:p> <jats:p>Contact: iwasaki@cb.k.u-tokyo.ac.jp</jats:p>