C 4 photosynthesis typically requires two specialized leaf cell types, bundle sheath (bs) and mesophyll (mp), which provide the foundation for this highly efficient carbon assimilation pathway. In leaves of Flaveria bidentis, a dicotyledonous C 4 plant, ribulose 1,5-bisphosphate carboxylase (rubisco) accumulates only in bs cells surrounding the vascular centers and not in mp cells. This is in contrast to the more common C 3 plants, which accumulate rubisco in all photosynthetic cells. Many previous studies have focused on transcriptional control of C 4 cell type-specificity; however, post-transcriptional regulation has also been implicated in the bs-specific expression of genes encoding the rubisco subunits. In this current study, a biolistic leaf transformation assay has provided direct evidence that the 5-and 3-untranslated regions (UTRs) of F. bidentis FbRbcS1 mRNA (from a nuclear gene encoding the rubisco small subunit), in themselves, confer strong bs cell-specific expression to gfpA reporter gene transcripts when transcribed from a constitutive CaMV promoter. In transformed leaf regions, strong bs cell-specific GFP expression was accompanied by corresponding bs cellspecific accumulation of the constitutively transcribed FbRbcS1 5-UTR-gfpA-3-UTR mRNAs. Control constructs lacking any RbcS mRNA sequences were expressed in all leaf cell types. These findings demonstrate that characteristic cell type-specific FbRbcS1 expression patterns in C 4 leaves can be established entirely by sequences contained within the transcribed UTRs of FbRbcS1 mRNAs. We conclude that selective transcript stabilization (in bs cells) or degradation (in mp cells) plays a key role in determining bs cell-specific localization of the rubisco enzyme.Plants that utilize the highly efficient C 4 pathway of carbon fixation typically possess a Kranz-type leaf anatomy that consists of two distinct photosynthetic cell types (1-4). Bundle sheath (bs) 3 cells occur as a layer around each leaf vein, while mesophyll (mp) cells occur in one or more layers surrounding the vascular associated rings of bs cells. This specialized leaf anatomy compartmentalizes two sets of photosynthetic reactions that make up the C 4 pathway. In the C 4 dicot Flaveria bidentis, atmospheric CO 2 entering via the leaf stomata is initially incorporated into four carbon acids by phosphoenolpyruvate carboxylase, an enzyme found only in the leaf mp cells. The C 4 acids diffuse from mp cells to bs cells, where they are decarboxylated by a photosynthetic malic enzyme. Within bs chloroplasts, the released CO 2 is incorporated into the Calvin cycle by ribulose 1,5-bisphosphate carboxylase/oxygenase (rubisco), the primary enzyme of photosynthetic carbon fixation. The specialized anatomical framework and compartmentalized reactions of the C 4 pathway function as a "CO 2 pump" that concentrates CO 2 in bs cells, where rubisco is specifically localized (1, 2, 5, 6). As a result, the carboxylase activity of this enzyme is increased, while its oxygenase activity, which can decrease phot...