Figure 5. Effect of TFIIAγ on rice-Xoc interaction.

(A) Interactions of myc-labelled TFB regions of TALEs from Xoc RH3 and FLAG-labelled rice TFIIAγs in N. benthamiana leaf cells analysed by CoIP assays. Proteins before (input) and after immunoprecipitation (IP) were detected with anti-myc and anti-FLAG antibodies. (B) TFIIAγ5-RNAi but not TFIIAγ1-RNAi plants showed enhanced resistance to Xoc strains. Each bar represents mean (total 30 to 35 leaves from five plants) ± standard deviation. b, significant difference between wild-type and transgenic plants after infection of a strain at p<0.01. (C) Mutation of TFIIAγ5 (rice line IRBB5) reduced expression of disease susceptibility gene OsSULTR3;6 after infection. Each bar represents mean (three replicates) ± standard deviation. b, significant difference between IR24 and IRBB5 at p<0.01.

DOI: http://dx.doi.org/10.7554/eLife.19605.021

Figure 5—figure supplement 1. Southern hybridization analysis of TALE genes in different Xanthomonas species.

DNA was digested with SphI and probed with the 2.9-kb SphI fragment of TALE gene pthXo1 from Xoo strain PXO99. The size positions of DNA markers are indicated at left. Xac, Xanthomonas axonopodis pv. citri; Xcv, Xanthomonas euvesicatoria; Xoc, Xanthomonas oryzae pv. oryzicola; Xcc, Xanthomonas campestris pv. camperstris. The Xcc strain 8004 is TALE-free (Qian et al., 2005, Genome Res. 15:757-767).

DOI: http://dx.doi.org/10.7554/eLife.19605.022

Figure 5—figure supplement 2. Analysis of interactions between Xoc TALEs and rice TFIIAγs.

The physical interactions between TFB regions of TALEs from Xoc RH3 and TFIIAγs were assessed by growth of yeast cells on synthetic defined premixes (SD) medium lacking leucine (L), tryptophan (W), histidine (H), and adenine (A). Among the 18 TFB regions, the sequences of TFB11, TFB15, TFB23, TFB24, and TFB27 from RH3 were the same as the TFB regions of Tal9b, Tal11a, Tal5b, Tal3c, and Tal12 from sequenced Xoc strain BLS256, respectively. (A) The TFB regions from RH3 interacted with rice TFIIAγ5 analysed using yeast two-hybrid assay. (B) A randomly chosen 7 of the 18 TFB regions did not interact with TFIIAγ5V39E (the mutated TFIIAγ5), but three interacted with mutated rice TFIIAγ1^S47E analysed by yeast two-hybrid assay. (C) Mutation of TFIIA5 enhanced resistance to Xoc. Plants were inoculated with Xoc at the booting stage. Each bar represents mean (total 40 to 50 leaves from 5 plants) ± standard deviation. b, significant difference between IR24 and IRBB5 plants at p<0.01.

DOI: http://dx.doi.org/10.7554/eLife.19605.023