Arabidopsis atrbohD-3 and atrbohF-3 knock-out mutants show reduced non-host resistance and enhanced susceptibility to aphids.
(A) Myzus persicae (Mp) and M. cerasi (Mc) performance on Arabidopsis atrbohD-3 and atrbohF-3 knock-out lines. Four-week old plants were exposed to two adult aphids and nymph production was counted after 10 days. Average nymph production for M. persicae and M. cerasi was calculated from three independent replicated experiments, with 10 plants per replicate per treatment. (B) Graph shows R. padi aphid survival on control (Col-0) and Arabidopsis atrbohD-3 and atrbohF-3 knock-out lines over 6 days. Three independent biological replicates were carried out, with 10 plants per replicate. Error bars indicate standard error. (C) Rhopalosiphum padi (Rp) survival on Arabidopsis atrbohD-3 and atrbohF-3 mutants and Col-0 wild-type plants between 3 and 4 days of the experiment. The two-tailed Student's t-test was used for statistical analyses (*** indicates p-value<0.001). Error bars indicate standard error. (D) ROS levels in atrbohD-3 and atrbohF-3 mutants during host and non-host interactions with aphids. Images were taken 3 and 24 hours after aphid exposure using a laser confocal microscope and processed in ImageJ to generate graph bars representing relative fluorescence to the control treatment (no aphids). Graph indicates fluorescence the relative fluorescence of leaf samples exposed to aphids compared to a no aphid control (indicated by C). Average relative ratios are based on 5 different leaf samples per treatment. Three independent replicated experiments, with 5 leaf samples per treatment per replicate, were performed with similar results. Graph shows results of one of the three replicates. Additional replicates are shown in S10 Fig.
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