Ribulose-1,5-bisphosphate carboxylase/oxygenase, the key enzyme of photosynthetic carbon fixation, is able to accept both O 2 and CO 2 as substrates. When it fixes O 2 , it produces 2-phosphoglycolate, which is detoxified by photorespiration and recycled to the Calvin-Benson-Bassham cycle. To complete photorespiration, metabolite transport across three organelles, chloroplasts, peroxisomes, and mitochondria, is necessary through transmembrane transporters. In rice (Oryza sativa) little is known about photorespiratory transmembrane transporters. Here, we identified the rice plastidic glycolate/glycerate translocator 1 (OsPLGG1), a homolog of Arabidopsis PLGG1. OsPLGG1 mutant lines, osplgg1-1, osplgg1-2, and osplgg1-3, showed a growth retardation phenotype, such as pale green leaf, reduced tiller number, and reduced seed grain weight as well as reduced photosynthetic carbon reduction rate due to low activities of photosystem I and II. The plant growth retardation in osplgg1 mutants was rescued under high CO 2 condition. Subcellular localization of OsPLGG1-GFP fusion protein, along with its predicted Nterminal transmembrane domain, confirmed that OsPLGG1 is a chloroplast transmembrane protein. Metabolite analysis indicated significant accumulation of photorespiratory metabolites, especially glycolate and glycerate, which have been shown to be transported by the Arabidopsis PLGG1, and changes for a number of metabolites which are not intermediates of photorespiration in the mutants. These results suggest that OsPLGG1 is the functional plastidic glycolate/glycerate transporter, which is necessary for photorespiration and growth in rice.

Glycoside hydrolase family1 β-glucosidases play a variety of roles in plants, but their in planta functions are largely unknown in rice (Oryza sativa). In this study, the biological function of Os12BGlu38, a rice β-glucosidase, expressed in bicellular to mature pollen, was examined. Genotype analysis of progeny of self-fertilized heterozygous Os12BGlu38 T-DNA mutant, os12bglu38-1, found no homozygotes and a 1:1 ratio of wild type (WT) to heterozygotes. Reciprocal cross analysis demonstrated that Os12BGlu38 deficiency cannot be inherited through the male gamete. In cytological analysis, the mature mutant pollen appeared shrunken and empty. Histochemical staining and transmission electron microscopy showed that mutant pollen lacked intine cell wall, which was rescued by introduction of WT Os12BGlu38 genomic DNA. Metabolite profiling analysis revealed that cutin monomers and waxes, the components of the pollen exine layer, were increased in anthers carrying pollen of os12bglu38-1 compared with WT and complemented lines. Os12BGlu38 fused with green fluorescent protein was localized to the plasma membrane (PM) in rice and tobacco. Recombinant Os12BGlu38 exhibited β-glucosidase activity on the universal substrate p-nitrophenyl β-d-glucoside and some oligosaccharides and glycosides. These findings provide evidence that function of a PM-associated β-glucosidase is necessary for proper intine development.

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Conjugation of phytohormones with glucose is a means of modulating their activities, which can be rapidly reversed by the action of β-glucosidases. Evaluation of previously characterized recombinant rice β-glucosidases found that nearly all could hydrolyze abscisic acid glucose ester (ABA-GE). Os4BGlu12 and Os4BGlu13, which are known to act on other phytohormones, had the highest activity. We expressed Os4BGlu12, Os4BGlu13 and other members of a highly similar rice chromosome 4 gene cluster (Os4BGlu9, Os4BGlu10 and Os4BGlu11) in transgenic Arabidopsis. Extracts of transgenic lines expressing each of the five genes had higher β-glucosidase activities on ABA-GE and gibberellin A4 glucose ester (GA4-GE). The β-glucosidase expression lines exhibited longer root and shoot lengths than control plants in response to salt and drought stress. Fusions of each of these proteins with green fluorescent protein localized near the plasma membrane and in the apoplast in tobacco leaf epithelial cells. The action of these extracellular β-glucosidases on multiple phytohormones suggests they may modulate the interactions between these phytohormones.