Abstract: Chloroplast genetic engineering allows site-specific modifications via two homologous recombination events. Comparing with nuclear genetic engineering, it is more efficient and much safer for exogenous gene expression. Chloroplast genetic engineering was most widely studied in the model plant, tobacco. In this review, we summarized the research progress of tobacco chloroplast genetic engineering in improving photosynthetic efficiency, resistance and quality. In addition, this article also discussed that biomass enzymes, secondary metabolites, vaccines and medicinal proteins were expressed in bioreactor by chloroplast genetic engineering. Based on above analysis, the research and application prospects of chloroplast genetic engineering were proposed.