Search

Summary Reactive oxygen species (ROS) play an important role during host–pathogen interactions and are often an indication of induced host defence responses. In this study, we demonstrate for the first time that Puccinia triticina (Pt) generates ROS, including superoxide, H2O2 and hydroxyl radicals, during wheat infection. Through pharmacological inhibition, we found that ROS are critical for both Pt urediniospore germination and pathogenic development on wheat. A comparative RNA‐Seq analysis of different stages of Pt infection process revealed 291 putative Pt genes associated with the oxidation–reduction process. Thirty‐seven of these genes encode known proteins. The expressions of five Pt genes, including PtNoxA, PtNoxB, PtNoxR, PtCat and PtSod, were subsequently verified using RT‐qPCR analysis. The results show that the expressions of PtNoxA, PtNoxB, PtNoxR, PtCat and PtSod are up‐regulated during urediniospore germination. In comparison, the expressions of PtNoxA, PtNoxB, PtNoxR and PtCat are down‐regulated during wheat infection from 12 to 120 h after inoculation (HAI), whereas the expression of PtSod is up‐regulated with a peak of expression at 120 HAI. We conclude that ROS are critical for the full virulence of Pt and a coordinate down‐regulation of PtNox genes may be important for successful infection in wheat.

Thatcher near-isogenic lines (NILs) of wheat carrying resistance gene Lr2a, Lr3, LrB or Lr9 were inoculated with Puccinia triticina races of virulence phenotype BBBD, MBDS, SBDG and FBDJ. Puccinia triticina infection structures were analysed under the fluorescence microscope over a course of 14 days after inoculation (dai). The relative proportion of P. triticina and wheat genomic DNA in infected leaves was estimated with a semiquantitative multiplex PCR analysis using P. triticinaand wheat-specific primers. The occurrence of a hypersensitive response (HR), cellular lignification and callose deposition in inoculated plants was investigated microscopically. In interactions producing highly resistant infection type (IT) ‘0;’, a maximum of two haustorial mother cells per infection site were produced, and there was no increase in the proportion of P. triticina genomic DNA in infected leaves, indicating the absence of P. triticina growth. In comparison, sizes of P. triticina colonies increased gradually in interactions producing moderately resistant IT ‘1’ and ‘2’, with the highest proportion of P. triticina genomic DNA found in leaves sampled at 14 dai. In interactions producing susceptible IT ‘3‐4’, the highest proportion of P. triticina genomic DNA was found in leaves sampled at 10 dai (45AE5‐51AE5%). HR and cellular lignification were induced in interactions producing IT ‘0;’ and ‘1’ at 1 dai but they were not observed in interactions producing IT ‘2’ until 2 dai. No HR or cellular lignification were induced in interactions producing susceptible IT ‘3‐4’. Furthermore, a strong deposition of callose was induced in Lr9 +B BBD andLr9 + FBDJ (IT ‘0;’), whereas this defence response was not induced in resistant or susceptible interactions involving Lr2a, Lr3 or LrB, indicating that Lr9 mediated resistance was different from that conditioned by Lr2a, Lr3 or LrB.

Defence responses were investigated in two potato cultivars with different levels of resistance to late blight, Russet Burbank (susceptible) and Kennebec (moderately resistant), after inoculation with single isolates representing Phytophthora infestans genotypes US-1 (previously predominant, mildly aggressive) and US-8 (currently predominant, highly aggressive). The accumulation of brown lignin-like materials and an increase in the cell wall affinity to trypan blue 24 h after inoculation were observed in cv. Kennebec inoculated with US-1, but not in Kennebec inoculated with US-8, or cv. Russet Burbank inoculated with either US-1 or US-8. The expression of PAL-1, HMG-2, PR-1 and PR-5 was investigated in three leaf strata (local, proximal and distal) and at different times after inoculation, using SYBR real-time RT-PCR. The activation of these defence-related genes was affected not only by P. infestans genotype, but also by the potato cultivar and the proximity to the inoculation site. These genes were up-regulated earlier in Kennebec than in Russet Burbank and in response to US-1 than to US-8. Over all, the earliest and strongest up-regulation of these genes occurred in Kennebec inoculated with US-1. Furthermore, PAL-1 and HMG-2 were down-regulated at the site of infection while such down-regulation was not observed for PR-1 or PR-5. In parallel, the accumulation level and location of phenolics and rishitin matched those of PAL-1 and HMG-2 transcripts, respectively. These results strongly suggest that changes in either the activation or suppression of defence responses by the pathogen shape the level of susceptibility of potato cultivars to late blight.