Plastid-expressed choline monooxygenase gene improves salt and drought tolerance through accumulation of glycine betaine in tobacco

Glycine betaine (GlyBet), a quaternary ammonium compound, functions as an osmoprotectant in many organisms including plants. Previous research has shown that over-expression of enzymes for GlyBet biosynthesis in transgenic plants improved abiotic stress tolerance, but so far no study on the effects of plastid-expression of choline monooxygenase, the enzyme that catalyzes the conversion of choline into betaine aldehyde, has been reported. In the present study, tobacco (Nicotiana tabacum L. cv Wisconsin 38) plants were transformed with a gene for choline monooxygenase (BvCMO) from beet (Beta vulgaris) via plastid genetic engineering. Transplastomic plants constitutively expressing BvCMO under the control of the ribosomal RNA operon promoter and a synthetic T7 gene G10 leader were able to accumulate GlyBet in leaves, roots and seeds, and exhibited improved tolerance to toxic level of choline and to salt/drought stress when compared to wild type plants. Transplastomic plants also demonstrated higher net photosynthetic rate and apparent quantum yield of photosynthesis in the presence of 150 mM NaCl. Salt stress caused no significant change on the maximal efficiency of PSII photochemistry (Fv/Fm) in both wild type and transplastomic plants, but a decrease in the actual efficiency of PSII (PhiPSII) was observed, and such a decrease was much greater in wild type plants. Our results demonstrate the feasibility of improving salt and drought tolerance in plants through plastid transformation with BvCMO gene.