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Background  In the model single-cell C 4  plant  Bienertia sinuspersici , chloroplast- and nuclear-encoded photosynthetic enzymes, characteristically confined to either bundle sheath or mesophyll cells in Kranz-type C 4  leaves, all occur together within individual leaf chlorenchyma cells. Intracellular separation of dimorphic chloroplasts and key enzymes within central and peripheral compartments allow for C 4  carbon fixation analogous to NAD-malic enzyme (NAD-ME) Kranz type species. Several methods were used to investigate dimorphic chloroplast differentiation in  B. sinuspersici .    Results  Confocal analysis revealed that Rubisco-containing chloroplasts in the central compartment chloroplasts (CCC) contained more photosystem II proteins than the peripheral compartment chloroplasts (PCC) which contain pyruvate,Pi dikinase (PPDK), a pattern analogous to the cell type-specific chloroplasts of many Kranz type NAD-ME species. Transient expression analysis using GFP fusion constructs containing various lengths of a  B. sinuspersici  Rubisco small subunit ( RbcS ) gene and the transit peptide of PPDK revealed that their import was not specific to either chloroplast type. Immunolocalization showed the  rbcL -specific mRNA binding protein RLSB to be selectively localized to the CCC in  B. sinuspersici , and to Rubisco-containing BS chloroplasts in the closely related Kranz species  Suaeda taxifolia.  Comparative fluorescence analyses were made using redox-sensitive and insensitive GFP forms, as well comparative staining using the peroxidase indicator 3,3-diaminobenzidine (DAB), which demonstrated differences in stromal redox potential, with the CCC having a more negative potential than the PCC.    Conclusions  Both CCC RLSB localization and the differential chloroplast redox state are suggested to have a role in post-transcriptional  rbcL  expression.

Exploring mechanisms linked to differentiation and function of dimorphic chloroplasts in the single cell C4 species Bienertia sinuspersici