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. 2019 Feb 8;20(4):599–608. doi: 10.1111/mpp.12779

Figure 6.

Figure 6

Interaction between OsRac1 and AvrPiz‐t/LF‐AvrPiz‐t and suppression of Rac1‐mediated reactive oxygen species (ROS) accumulation by AvrPiz‐t and LF‐AvrPiz‐t in Nicotiana benthamiana. (a) Luciferase complementation imaging (LCI) assay for the interaction between OsRac1 and AvrPiz‐t/LF‐AvrPiz‐t. OsRac1‐Nluc was co‐expressed with AvrPiz‐t‐Cluc or LF‐AvrPiz‐t‐Cluc, whereas Nluc‐empty vector/AvrPiz‐t‐Cluc, Nluc‐empty vector/LF‐AvrPiz‐t‐Cluc and OsRac1‐Nluc/Cluc‐empty vector were used as negative controls. OsRac1‐Nluc/OsRBOHB‐Cluc was used as a positive control. The numbers in parentheses indicate the number of leaf spots with positive luminescence signals among all of the infiltrated leaves. (b) Relative luciferase (LUC) activity of the interaction between OsRac1 and AvrPiz‐t/LF‐AvrPiz‐t. Three biological replicates with 18 N. benthamiana leaves were measured for the luciferase activity using the Image Lab quantity tool. (c) Rac1‐mediated ROS production in N. benthamiana leaves expressing AvrPiz‐t, LF‐AvrPiz‐t or empty vector with OsRac1. Leaf discs of each infiltrated spot on the same leaves were treated with 8 nm chitin. ROS accumulation was measured using luminol‐based chemiluminescence for 15 min. Values are means ± standard error (SE) (n = 3). RLU, relative light unit.