FIG 7 .

Effect of Agrobacterium-mediated transient expressions of RipAY and RipAY E216Q in N. benthamiana. (A) Leaves were infiltrated with A. tumefaciens strain GV3101 harboring the binary vector carrying the effector genes or empty vector. “Mock” represents the infiltration of buffer only. Photographs were taken 2 days after agroinfiltration. (B) GSH level in the infiltrated leaves. The GSH levels were measured 2 days after infiltration. The data are means ± SD from three independent experiments. (C) Subcellular localization of RipAY. GFP, GFP-RipAY, and GFP-RipAY E216Q were each transiently expressed in leaves by agroinfiltration. Two days after infiltration, protoplasts were prepared from infiltrated leaves, and the subcellular localization of GFP fluorescence was monitored by a confocal laser scanning microscopy. (D) ROS production in the infiltrated leaves. Two days after infiltration, leaf disks were treated with the flg22 elicitor at 100 µM, and the flg22-triggered ROS production was measured as photon counts for 120 min. The data are means ± SD from eight independent leaf disks. (E) Expression of defense-related genes in the infiltrated leaves. Total RNAs were extracted from leaves treated with flg22 at the indicated times in minutes, and the expression of PTI marker genes were analyzed by RT-qPCR.