We introduced the Escherichia coli glycolate catabolic pathway into Arabidopsis thaliana chloroplasts to reduce the loss of fixed carbon and nitrogen that occurs in C3 plants when phosphoglycolate, an inevitable by-product of photosynthesis, is recycled by photorespiration. Using step-wise nuclear transformation with five chloroplast-targeted bacterial genes encoding glycolate dehydrogenase, glyoxylate carboligase and tartronic semialdehyde reductase, we generated plants in which chloroplastic glycolate is converted directly to glycerate. This reduces, but does not eliminate, flux of photorespiratory metabolites through peroxisomes and mitochondria. Transgenic plants grew faster, produced more shoot and root biomass, and contained more soluble sugars, reflecting reduced photorespiration and enhanced photosynthesis that correlated with an increased chloroplastic CO2 concentration in the vicinity of ribulose-1,5-bisphosphate carboxylase/oxygenase. These effects are evident after overexpression of the three subunits of glycolate dehydrogenase, but enhanced by introducing the complete bacterial glycolate catabolic pathway. Diverting chloroplastic glycolate from photorespiration may improve the productivity of crops with C3 photosynthesis.
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This work was supported by a PhD scholarship from the Egyptian government to R.K. and grants from the Deutsche Forschungsgemeinschaft and Bayer CropScience to C.P. Thanks to Martin Parry, Alf Keys, Rainer Häusler, Veronica Maurino, Susanne von Caemmerer, John Evans, Margrit Frentzen, Dagmar Weier, Thomas Rademacher, Burkhard Schmidt and Nikolaus Schlaich for helpful discussion of this work and technical support.
The authors declare no competing financial interests.
Multiplex PCR for the detection of plants containing all five open reading frames for the installation of the E. coli glycolate pathway in Arabidopsis chloroplasts. (PDF 41 kb)
Additional growth parameters of wild-type and transgenic lines. (PDF 16 kb)
Glycine and serine concentrations in leaves of wild-type and transgenic lines. (PDF 10 kb)
Typical measuring profile for the determination of the postillumination CO2 burst (PIB). (PDF 24 kb)
Typical measuring profile for the determination of the CO2 compensation point Γ* and the dark respiration rate in the light (Rd). (PDF 77 kb)
Genomic integration sites of the T-DNAs containing the listed constructs. (PDF 9 kb)
Generation and selection of transgenic lines used in this study. (PDF 15 kb)
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Kebeish, R., Niessen, M., Thiruveedhi, K. et al. Chloroplastic photorespiratory bypass increases photosynthesis and biomass production in Arabidopsis thaliana. Nat Biotechnol 25, 593–599 (2007). https://doi.org/10.1038/nbt1299
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