Plant Physiol, September 2000, Vol. 124, pp. 163-172

Changes in Photosynthetic Carbon Flow in Transgenic Rice Plants That Express C4-Type Phosphoenolpyruvate Carboxykinase from Urochloa panicoides

Plant Breeding Genetics Research Laboratory, Japan Tobacco Inc., 700 Higashibara, Toyoda, Iwata, Shizuoka 438-0802, Japan (S.S., N.M., M.A.); and Department of Biochemistry and Molecular Biology, James Cook University, Townsville, Queensland 4811, Australia (J.N.B.)

A cDNA encoding phosphoenolpyruvate carboxykinase (PCK) of Urochloa panicoides (a PCK-type C4 plant) was expressed in rice (Oryza sativa cv Tsukinohikari) plants under the control of the promoter of a maize (Zea mays) gene for phosphoenolpyruvate carboxylase or pyruvate, orthophosphate dikinase with the transit peptide of the small subunit of Rubisco. Crude extracts prepared from the green leaves of transgenic plants had high PCK activity and the newly expressed PCK was localized in chloroplasts. In labeling experiments with ¹⁴CO₂ up to 20% of the radioactivity was incorporated into 4C compounds (malate, oxaloacetate, and aspartate) in excised leaves of transgenic plants, as compared with about 1% in excised leaves of control plants. There was a positive correlation between PCK activity and the extent of labeling of 4C compounds. When L-[4-¹⁴C]malate was fed to excised leaves the extent of incorporation of radioactivity into sucrose was 3-fold greater in transgenic plants than in control plants and the level of radiolabeled aspartate was significantly lower in transgenic plants. These results indicate that the ectopic expression of PCK in rice chloroplasts was able partially to change the carbon flow in mesophyll cells into a C4-like photosynthetic pathway. Such a strategy appears to provide a possible method for enhancing the photosynthetic capacity of C3 plants.