Enhanced antioxidant defence plays an essential role in plant survival under stress conditions. However, excessive antioxidant activity sometimes suppresses the signal necessary for the initiation of the desired biological reactions. One such example is microspore embryogenesis (ME)—a process of embryo-like structure formation triggered by stress in immature male gametophytes. The study focused on the role of reactive oxygen species and antioxidant defence in triticale (×Triticosecale Wittm.) and barley (Hordeum vulgare L.) microspore reprogramming. ME was induced through various stress treatments of tillers and its effectiveness was analysed in terms of ascorbate and glutathione contents, total activity of low molecular weight antioxidants and activities of glutathione–ascorbate cycle enzymes. The most effective treatment for both species was a combination of low temperature and exogenous application of 0.3 M mannitol, with or without 0.3 mM reduced glutathione. The applied treatments induced genotype-specific defence responses. In triticale, both ascorbate and glutathione were associated with ME induction, though the role of glutathione did not seem to be related to its function as a reducing agent. In barley, effective ME was accompanied by an accumulation of ascorbate and high activity of enzymes regulating its redox status, without direct relation to glutathione content.

The plant hormones cytokinins affect a various array of plant growth and development processes as well as responses to biotic and abiotic stresses. In this study, the opposite effect of two different cytokinins kinetin (N⁶-furfuryladenine) and benzyladenine (BA) on development and on the tolerance of Arabidopsis and tobacco plants to virus, bacteria, and fungi infection was reported. Treatments of Arabidopsis and tobacco seedlings with saturated solutions of BA inhibited plant progress, while treatments with saturated water solution of kinetin promoted plant development. Furthermore, BA pre-treatments strongly reduced the number of TMV (Tobacco mosaic virus) lesions on tobacco and the tissue damage caused by the incompatible Pseudomonas bacteria on Arabidopsis and tobacco leaves. Similarly, BA pre-treatment significantly reduced the necrotic disease symptoms of Botrytis cinerea infection. Kinetin pre-treatments had a much weaker or no protective effect on the damage caused by the above pathogens. Accordingly, Arabidopsis gene expression profiles after treatments also showed that the two cytokinins have different effects on several plant processes. The gene expression results supported the more robust effect of BA, which up and downregulated more than 2000 genes, while only 436 genes were influenced by kinetin treatment. It is noteworthy that BA and kinetin treatment changed gene expressions in the same direction only in a relatively few cases (73 upregulated and 70 downregulated genes), and even 28 genes were regulated into the opposite directions by BA and kinetin. Both treatments had a strong effect on auxin and gibberellin-related genes, but only BA had a significant effect on cytokinin-induced processes. While kinetin exclusively activated the flavonoid synthesis genes, BA affected more significantly protein synthesis, photosynthesis, and plant defence-related genes. In conclusion, BA solution had sometimes the opposite and generally a much stronger effect than kinetin solution not only on the development and on biotic stress tolerance of tobacco and Arabidopsis plants but also on the gene expressions. The stronger protective effect of BA to necrotic stresses is probably due to its stronger senescence inhibitory effect on plant tissues, as supported by the stronger chlorophyll retardation of the BA-treated leaves.

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 physiological state, i.e., senescence or juvenility, of plants and plant organs can have strong effect on their reactions to pathogen attacks. This effect is mainly expressed as changes in the severity of disease symptoms. Generally, necrotrophic pathogens cause more severe symptoms on senescent than on juvenile plants, while biotrophs prefer juvenile tissues. Several factors of senescence have opposite effect on the two pathogen groups, such as decreased photosynthesis, decreased antioxidant capacity, remobilization of nutrients, changes in plant hormonal network, and in fluidity of cell membranes. Furthermore, senescent tissues are less tolerant to toxins and to cell-wall-degrading enzymes. On the other hand, pathogen infection itself has significant effect on the physiology of plants depending on the lifestyle of the pathogen and on the compatibility or incompatibility of the interaction with the plant. There are several possibilities to manipulate the physiological state of plants in order to improve their biotic and abiotic stress tolerance, such as removal of the terminal bud or high doses of nitrogen, external application of cytokinins or of inhibitors of ethylene action, as well as by spontaneous or directed mutation, in vitro selection, or manipulation by various transgenic approach. Even application of mycorrhiza can inhibit the senescence process of plants and improve their tolerance to stresses.

Heat shock treatment of near isogenic barley lines induced susceptibility against powdery mildew (Blumeria graminis f. sp. hordei, Bgh). When barley lines were immersed into hot water (48–49 °C) for 20 seconds one day before inoculation with Bgh race A6, the heat treatment increased susceptibility in susceptible barley cv. Ingrid and in its near-isogenic barley lines carrying different effective resistance genes. Microscopic investigations indicated vigorous development of the pathogen not only on heat treated susceptible Ingrid and resistant Mla, but also on Mlg-resistant and even mlo-resistant lines. However, when longer heat stress was used, infection density increased gradually on the susceptible Ingrid leaves, and the 40–50 sec heat treatment induced the development of visible powdery mildew colonies even on mlo leaves. Heat stress significantly increased leakage of ions from leaf segments from all barley lines with or without specific resistance genes and caused a late decrease of SOD and a slight increase in CAT enzyme activities, which correlated with the slightly down-regulated levels of hydrogen peroxide in the heat treated barley leaves. Significant increase of RNase activities was found after heat stress, and there was a slight degradation of total DNA as a consequence of heat pretreatment in all barley lines