High ammonium supply impairs photosynthetic efficiency in rice exposed to excess light.

Mechanisms involving ammonium toxicity, excess light, and photosynthesis are scarcely known in plants. We tested the hypothesis that high NH4+ supply in presence of high light decreases photosynthetic efficiency of rice plants, an allegedly tolerant species. Mature rice plants were previously supplied with 10 mM NH4+ or 10 mM NO3- and subsequently exposed to 400 µmol m-2 s-1 (moderate light-ML) or 2000 µmol m-2 s-1 (high light-HL) for 8 h. HL greatly stimulated NH4+ accumulation in roots and in a minor extent in leaves. These plants displayed significant delay in D1 protein recovery in the dark, compared to nitrate-supplied plants. These responses were related to reduction of both PSII and PSI quantum efficiencies and induction of non-photochemical quenching. These changes were also associated with higher limitation in the donor side and lower restriction in the acceptor side of PSI. This later response was closely related to prominent decrease in stomatal conductance and net CO2 assimilation that could have strongly affected the energy balance in chloroplast, favoring ATP accumulation and NPQ induction. In parallel, NH4+ induced a strong increase in the electron flux to photorespiration and, inversely, it decreased the flux to Rubisco carboxylation. Overall, ammonium supply negatively interacts with excess light, possibly by enhancing ammonium transport towards leaves, causing negative effects on some photosynthetic steps. We propose that high ammonium supply to rice combined with excess light is capable to induce strong delay in D1 protein turnover and restriction in stomatal conductance, which might have contributed to generalized disturbances on photosynthetic efficiency.

Modulation of genes related to specific metabolic pathways in response to cytosolic ascorbate peroxidase knockdown in rice plants.

As a central component of the hydrogen peroxide detoxifying system in plant cells, ascorbate peroxidases (APX) play an essential role in the control of intracellular reactive oxygen species (ROS) levels. To characterise the function of cytosolic APX isoforms (OsAPX1 and OsAPX2) in the mechanisms of plant defence, OsAPX1/2 knockdown rice plants were previously obtained. OsAPX1/2 knockdown plants (APx1/2s) exhibited a normal phenotype and development, even though they showed a global reduction of APX activity and increased hydrogen peroxide accumulation. To understand how rice plants compensate for the deficiency of cytosolic APX, expression and proteomic analyses were performed to characterise the global expression pattern of the APx1/2s mutant line compared with non-transformed plants. Our results strongly suggest that deficiencies in cytosolic APX isoforms markedly alter expression of genes associated with several key metabolic pathways, especially of genes involved in photosynthesis and antioxidant defence. These metabolic changes are compensatory because central physiological processes such as photosynthesis and growth were similar to non-transformed rice plants. Our analyses showed modulation of groups of genes and proteins related to specific metabolic pathways. Among the differentially expressed genes, the largest number corresponded to those with catalytic activity. Genes related to oxidative stress, carbohydrate metabolism, photosynthesis and transcription factor-encoding genes were also modulated. These results represent an important step toward understanding of the role played by cytosolic APX isoforms and hydrogen peroxide in the regulation of metabolism by redox modulation in monocots.