Glycine betaine (GB) plays a crucial role in plant response to abiotic stress, and its accumulation in chloroplasts is more effective than in the cytosol in improving the resistance of transgenic plants. Here, we report that the codA gene from Arthrobacter globiformis, which encodes a choline oxidase catalysing the conversion of choline to GB, was successfully introduced into the plastid genome of potato (Solanum tuberosum L.). Transgenic plants with plastid expression of codA showed increased tolerance to low temperature stress compared with the wild type (WT). Further studies revealed that under low temperature stress condition, transgenic plants presented a significantly higher photosynthetic performance by regulating the electron transport and energy distribution in PSII, and higher antioxidant enzyme activities and lower O2- and H2O2 accumulation than did the WT plants. A higher expression of the COR genes was also observed in transgenic plants. Our results suggest that chloroplast biosynthesis of GB could be an effective strategy for the engineering of plants with increased resistance to low temperature stress.