Requirement of predicted catalytic residues of AvrRxo1 for toxic or growth-suppressive effects in prokaryotes and eukaryotes and for induction of HR in Rxo1 rice.
(A) E. coli transformed with pDEST-AvrRxo1, pDEST-AvrRxo1-D193T, pDEST-AvrRxo1-T167N, and pDESTcv grown on (-IPTG) and inducing (+IPTG) media. (B) S. cerevisiae transformed with pESC-Trp-DEST-AvrRxo1 and catalytic site mutant derivatives pESC-Trp-DEST-D193T and -T167N on repressing (SD media amended with 2% glucose) and inducing (SD media amended with 2%/1% galactose/raffinose) media. (C) N. benthamiana leaf during transient Agrobacterium-mediated expression of HA-AvrRxo1, HA-AvrRxo1-D193T and HA-AvrRxo1-T167N. Leaf was imaged 50 hours after infiltration. Expression analysis shown in S1 Fig. (D) Leaves of transgenic rice variety Kitaake expressing Rxo1 were inoculated with derivatives of X. oryzae strain X11-5A carrying pHM1-AvrRxo1, pHM1-AvrRxo1-D193T, pHM1-AvrRxo1-T167N, or the empty vector pHM1. Appearance of water-soaking lesions or HR were recorded 5 days post infiltration.