Avr3b recognition in soybean is dependent on the PPlase activity of GmCYP1.

<p>(<b>A, B</b>) CsA suppresses Avr3b-triggered cell death in soybean leaves. <b>(A)</b> Soybean cultivars Williams (rps3b) and PRX146-36 (Rps3b) were transformed by co-bombardment with a plasmid mixture consisting of a β-glucuronidase (GUS) expression vector and Avr3b or empty vector (EV). Soybean leaves were sprayed 20 μM CsA solution immediately after bombardment and incubated for 2 days in darkness at 28°C. The leaves were then stained using X-gluc. Scale bars, 3mm. (<b>B</b>) Percentage rate of GUS-positive blue spots following bombardment with Avr3b compared with EV. Bars represent standard errors from four independent replicates. **, <i>t</i> test <i>P</i><0.01. (<b>C, D</b>) CsA could not suppress Avr1b-triggered cell death. (<b>C</b>) Avr1b was transiently expressed in soybean cultivars Williams (rps1b) and L77-1863 (Rps1b) by co-bombardment with GUS expression vector and Avr1b or EV. The leaves were incubated for 2 days in darkness at 28°C after bombardment, and then stained using X-gluc. Scale bars, 3mm. (<b>D</b>) Percentage rate of GUS-positive blue spots following bombardment with Avr1b compared with the empty vector. Bars represent standard errors from four independent replicates. **, <i>t</i> test <i>P</i><0.01. (<b>E, F</b>) Avr3b failed to induce Rps3b-mediated cell death in <i>GmCYP1</i>-silenced soybean leaves. (<b>E</b>) Avr3b was delivered into leaves of soybean cultivars Williams (rps3b) and PRX146-36 (Rps3b) together with a <i>GmCYP1</i>-RNAi construct by co-bombardment. The leaves were incubated for 2 days in darkness at 28°C after bombardment, and then stained using X-gluc. Scale bars, 3mm. EV was used as a negative control. (<b>F</b>) Percentage rate of GUS-positive blue spots following bombardment with Avr3b compared with the EV in the presence of <i>GmCYP1</i>-RNAi. Bars represent standard errors from four independent replicates. **, <i>t</i> test <i>P</i><0.01.