Subcellular localization and enzymatic properties of differentially expressed transketolase genes isolated from the desiccation tolerant resurrection plant Craterostigma plantagineum

The desiccation tolerant resurrection plant Craterostigma plantagineum encodes three classes of transketolase transcripts, which are distinguished by their gene structures and their expression patterns. One class, represented by tkt3, is constitutively expressed and two classes, represented by tkt7 and tkt10, are upregulated upon rehydration of desiccated C. plantagineum plants. The objective of this work was to characterize the differentially expressed transketolase isoforms with respect to subcellular localization and enzymatic activity. Using GFP fusion constructs and enzymatic activity assays, we demonstrate that C. plantagineum has novel forms of transketolase which localize not to the chloroplast, but mainly to the cytoplasm and which are distinct in the enzymatic properties from the transketolase enzymes active in the Calvin cycle or oxidative pentose phosphate pathway. A transketolase preparation from rehydrated leaves was able to synthesize the unusual C8 carbon sugar octulose when glucose-6-phosphate and hydroxy-pyruvate were used as acceptor and donor molecules in in vitro assays. This suggests that a transketolase catalyzed reaction is likely to be involved in the octulose biosynthesis in C. plantagineum.