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. 2016 May 3;39(5):426–438. doi: 10.14348/molcells.2016.0094

Fig. 5.

Fig. 5.

Inhibition of Os-NADP-ME2-3 attenuates PAMP-triggered ROS burst. (A) and (B), AVR-Pii specifically inhibits Os-NADP-ME2-3 activity. The activity of Os-NADP-ME2-3 alone or in the presence of AVR-Pii or AVR-Pii-MT, was determined at varying concentrations of NADP+ at a saturating concentration of L-Malate (3.5 mM) (A) and at varying concentrations of L-Malate at a saturating concentration of NADP+ (0.3 mM) (B) with fixed levels of other the substrates. Results are presented as mean ± SD of triplicate determinants. (C) Oxidative burst elicited by Chitin (8 nM) in HY and ΔOs-nadp-me2-3 leaf disc. Deletion of Os-NADP-ME2-3 inhibited PAMP-triggered ROS bursts. ROS generation was measured using luminol with a luminometer (SpectraMax L Microplate Reader). A total of 100 μl luminol, 1 μl horseradish peroxidase, and 8 nM chitin, and sterile water (as a control) were added to HY and ΔOs-nadp-me2-3 mutants. Values represent the mean ± SD (n = 16). The experiment was repeated three times with similar results.