Vacuolar invertase activity shapes photosynthetic stress response of Arabidopsis thaliana and stabilizes central energy supply

Stabilization of the central carbohydrate and energy metabolism plays a key role in plant stress response. As the primary photosynthetic products, carbohydrates are substrate for numerous metabolic and stress-protective reactions. Further, they have been shown to be involved in diverse signalling processes finally affecting and regulating plant stress response on a whole plant level. Sucrose metabolism is known to be central to many stress-related processes and enzymes catalysing its biosynthesis, transport and degradation have been shown to significantly impact stress resistance and acclimation output. However, due to the cyclic structure of sucrose metabolism involving sucrose cleavage in multiple compartments as well as energy-demanding re-synthesis via hexose phosphorylation, it is challenging to derive an unambiguous picture of its contribution to stress reactions. In the present study, a combined stress experiment comprising cold and high-light identified metabolism of sucrose and fumaric acid to significantly separate the stress response of a cold susceptible and a tolerant natural accession of Arabidopsis thaliana. Kinetic modelling and simulation of subcellular rates of invertase-driven sucrose cleavage revealed a contrasting picture between the susceptible and the tolerant accession pointing to an important role of vacuolar invertase during initial stress response. Using a T-DNA insertion mutant with a dramatically reduced invertase activity provided evidence for a central role of the enzyme in stabilizing photosynthesis and the central energy metabolism during freezing and high-light stress. Reducing vacuolar invertase activity to about 3% of the wild type resulted in a strong increase of ADP and ATP levels indicating a severe effect on cytosolic and plastidial energy balance. Together with a significant decrease of maximum quantum yield of photosystem II (Fv/Fm) these results suggest that vacuolar invertase activity stabilizes cytosolic energy metabolism by supplying hexose equivalents being phosphorylated in the cytosol. Finally, the accompanying ATP consumption is essential for cytosolic phosphate balance which directly affects photosynthetic performance by the supply of ADP being crucial for photosynthetic ATP production.