Development of C[sub.4] Biochemistry and Change in Expression of Markers for Photosystems I and II in the Single-Cell C[sub.4] Species, IBienertia sinuspersici/I

Bienertia sinuspersici is one of four identified terrestrial plants that perform C[sub.4] photosynthesis within a single chlorenchyma cell via the compartmentation of organelles and photosynthetic enzymes. The patterns of accumulation of key photosynthetic enzymes and transcripts in developing leaves were examined using immunolocalization and in situ hybridization. The polypeptides of Rubisco large subunit (RbcL) and pyruvate Pi dikinase (PPDK) accumulated equally in all chloroplasts before the formation of two intracellular cytoplasmic compartments: the central (CCC) and peripheral (PCC) cytoplasmic compartments. The differential accumulation of these enzymes was not completed until the leaf had reached maturity, indicating that the transition from C[sub.3] to C[sub.4] photosynthesis occurred during leaf maturation. In mature chlorenchyma cells, RbcL accumulated 20-fold higher in the CCC than in the PCC, while PPDK exhibited a concentration gradient that was the lowest in the chloroplasts in the central region of the CCC and the highest in PCC chloroplasts. The pattern of rbcL transcript accumulation followed that of its polypeptides in developing leaves, suggesting that the expression of this gene was likely controlled by transcriptional and/or post-transcriptional processes. Immunocytochemical results examining the distribution of photosystems I and II in the chloroplasts of chlorenchyma cells from mature leaves showed that PSII is more abundant in chloroplasts of the central compartment, whereas PSI is higher in those of the peripheral compartment. The quantitative real-time PCR results of rbcL, psbA, and psaB transcripts from the isolated chloroplasts of each compartment further supported this observation. Our results suggest that multiple levels of regulation play a role in controlling the differential accumulation of photosynthetic gene expression in the dimorphic chloroplasts of single-cell C[sub.4] species during leaf development.