Figure 6.
The production of reactive oxygen species and gene expression profiles of M. truncatula wild-type R108 and the stay-green (sgr) mutant to P. pachyrhizi. (A) 3,3-diaminobenzidine (DAB) staining for detection of hydrogen peroxide. Four-week-old M. truncatula wild-type R108 and sgr mutant were spray-inoculated with 1 × 105 spores/ml (0.001% Tween 20) of P. pachyrhizi, then incubated with DAB for 1, 2 and 4 days post-inoculation (dpi). (B) Expression profiles of genes involved in defense responses in M. truncatula inoculated with P. pachyrhizi. M. truncatula leaves spray-inoculated with P. pachyrhizi were sampled at 12, 24, 48 and 72 hours post-inoculation (hpi) or mock control (0 time). Expression profiles are presented for genes encoding Pathogenesis-related protein 3 (PR3), Pathogenesis-related protein 4 (PR4), pathogenesis-related protein 10 (PR10) and Hsr203J. The expression of genes was evaluated by RT-qPCR with gene-specific primer sets (Supplementary Table S10). The values represent the fold induction compared to mock control. Three biological replicates were used for each inoculation. The Ubiquitin gene was used as an internal control. Bars represent the means ± standard deviation (SD).