Earlier studies showed that the artificial elevation of endogenous glutathione (GSH) contents can markedly increase the resistance of plants against different viruses. On the other hand, salicylic acid (SA)-deficient NahG plants display enhanced susceptibility to viral infections. In the present study, the biochemical mechanisms underlying GSH-induced resistance were investigated in various tobacco biotypes displaying markedly different GSH and SA levels. The endogenous GSH levels of Nicotiana tabacum cv. Xanthi NN and N. tabacum cv. Xanthi NN NahG tobacco leaves were increased by infiltration of exogenous GSH or its synthetic precursor R-2-oxo-4-thiazolidine-carboxylic acid (OTC). Alternatively, we also used tobacco lines containing high GSH levels due to transgenes encoding critical enzymes for cysteine and GSH biosynthesis. We crossed Xanthi NN and NahG tobaccos with the GSH overproducer transgenic tobacco lines in order to obtain F₁ progenies with increased levels of GSH and decreased levels of SA. We demonstrated that in SA-deficient NahG tobacco the elevation of in planta GSH and GSSG levels either by exogenous GSH or by crossing with glutathione overproducing plants confers enhanced resistance to Tobacco mosaic virus (TMV) manifested as both reduced symptoms (i.e. suppression of hypersensitive-type localized necrosis) and lower virus titers. The beneficial effects of elevated GSH on TMV resistance was markedly stronger in NahG than in Xanthi NN leaves. Infiltration of exogenous GSH and OTC or crossing with GSH overproducer tobacco lines resulted in a substantial rise of bound SA and to a lesser extent of free SA levels in tobacco, especially following TMV infection. Significant increases in expression of pathogenesis related (NtPR-1a, and NtPRB-1b), and glutathione S-transferase (NtGSTtau, and NtGSTphi) genes were evident in TMV-inoculated leaves in later stages of pathogenesis. However, the highest levels of defense gene expression were associated with SA-deficiency, rather than enhanced TMV resistance. In summary, elevated levels of glutathione in TMV-infected tobacco can compensate for SA deficiency to maintain virus resistance. Our results suggest that glutathione-induced redox changes are important components of antiviral signaling in tobacco.

The present work focused on the characterization of some physiological mechanisms activated upon powdery mildew inoculation of the susceptible barley cultivar Ingrid and its near-isogenic lines (NILs) carrying various resistant genes (Mla, Mlg and mlo). After inoculation with Blumeria graminis f. sp. hordei (Bgh), measurements of leaf reflectance and chlorophyll a fluorescence were performed 3 and 7 day post-inoculation (dpi), while hormone assays were made 7 dpi. Bgh-inoculated resistant genotypes were characterized by lowered leaf reflectance parameters that correlated with carotenoids (CRI) and water content (WBI) in comparison to inoculated Ingrid. The PSII activity (i.e., Fv/Fm, ETo/CSm and P.I._ABS) strongly decreased in susceptible Ingrid leaves when the disease symptoms became visible 7 dpi. In Mla plants with visible hypersensitive spots the PSII activity decreased to a lesser extent. Inoculation resulted in a very slight decrease of photosynthesis at later stage of infection in Mlg plants, whereas in resistant mlo plants the PSII activity did not change. Chlorophyll a fluorescence measurements allowed presymptomatic detection of infection in Ingrid and Mla. Changes in the homeostasis of 22 phytohormones (cytokinins, auxins, gibberellins and the stress hormones JA, SA and ABA) in powdery mildew inoculated barley are discussed in relation to resistance against this biotrophic pathogen.

The monoterpenoid (S)-carvone was shown earlier to induce glutathione S-transferase (GST) activity and to attenuate necrotic lesion formation in Tobacco mosaic virus (TMV) inoculated, hypersensitively reacting tobacco Xanthi-nc plants. To explore structure-activity relationships, three monoterpenes and five monoterpenoids were tested for their ability to induce increased GST activity in tobacco leaf discs. (S)-Carvone proved to be the most potent inducer. (S)-Carvone treatments markedly up-regulated the expression of several tobacco GST genes belonging to the phi, tau and theta classes, particularly in the case of tau class GSTs. Furthermore, TMV inoculation slightly induced the transcript abundance of the GSTT2 gene. The expression of three catalase (CAT) genes was also examined in (S)-carvone-treated tobacco leaf discs. The expression of CAT1 was not influenced by (S)-carvone treatments, whereas that of CAT2 was significantly repressed. (S)-Carvone very weakly induced the transcription of the CAT3 gene. The effect of the opposite enantiomer, (R)-carvone on the expression of GST and CAT genes was similar to that of (S)-carvone. Our results suggest that the antioxidative reactions catalyzed by tau and theta class GSTs contribute to the attenuation of necrotic disease symptoms in TMV-inoculated tobacco plants