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Nitric oxide (NO) plays a key role in plant metabolism, signaling, defense, and development. However, a fundamental question arises how the NO message is converted into a physiological response. NO-related signaling may be attributed to various NO derivatives, collectively named reactive nitrogen species (RNS). An increasing body of evidence suggests that nitroxyl (HNO) as a one-electron reduced and protonated state of NO and nitrosonium (NO+), a one-electron oxidized form of NO, may be important factors. Thus, the impact of NO in plant biology should be re-evaluated in the light of chemical properties of these compounds as they are different from that of NO. Nitroxyl, unlike NO, can interact directly with thiols, prioritize ferric rather than ferrous heme proteins and it is resistant to scavenging by superoxides. Experimental data revealed that NO+ can facilitate S-nitrosylation, while it also appears to be a key intermediate in the trans-nitrosylation reaction. In contrast to NO, it may also take part in the degradation of SNOs (S-nitrosothiols). In turn, peroxynitrite (ONOO−) is a potent oxidant and nitrating agent, generated by the reaction of nitric oxide and superoxide in one of the most rapid reactions known in biology. Overproduction of ONOO− contributes to oxidative and nitrosative stress, however, in the physiological state or under low metabolic stress, peroxynitrite triggered NO-dependent signals behave as a potent modulator of the redox regulation in various cell transduction pathways.

The aim of the study was to examine cross-talk of exogenous nitric oxide (NO) and sucrose in the mechanisms of synthesis and accumulation of isoflavonoids in embryo axes of Lupinus luteus L. cv. Juno. It was verified whether the interaction of these molecules can modulate the defense response of axes to infection and development of the pathogenic fungus Fusarium oxysporum f. sp. lupini . Sucrose alone strongly stimulated a high level of genistein glucoside in axes pretreated with exogenous nitric oxide (SNP or GSNO) and non-pretreated axes. As a result of amplification of the signal coming from sucrose and GSNO, high isoflavonoids accumulation was observed (+Sn+GSNO). It needs to be stressed that infection in tissues pretreated with SNP/GSNO and cultured on the medium with sucrose (+Si+SNP/+Si+GSNO) very strongly enhances the accumulation of free isoflavone aglycones. In +Si+SNP axes phenylalanine ammonia-lyase activity was high up to 72 h. As early as at 12 h in +Si+SNP axes an increase was recorded in gene expression level of the specific isoflavonoid synthesis pathway. At 24 h in +Si+SNP axes a very high total antioxidant capacity dependent on the pool of fast antioxidants was noted. Post-infection generation of semiquinone radicals was lower in axes with a high level of sucrose than with a deficit.

Homocysteine (Hcy) is a naturally occurring intermediate metabolite formed during methionine metabolism. It has been well documented that its excess can be extremely toxic to mammalian, yeast and bacterial cells. In spite of the metabolic value of Hcy known for decades, the role of this amino acid in the plant response to stress has not been recognized yet. In the presented study, using potato plant (Solanum tuberosum L.) and Phytophthora infestans as a model system, the presence and tissue localization of Hcy in leaves was examined by an immunohistochemical method. The over-production of Hcy was more evidenced in the susceptible than in the resistant genotype of potato starting from 48 hpi. Furthermore, the elevated level of Hcy was correlated in time with the up-regulation of genes engaged in its biosynthesis, e.g. cystathionine β-lyase and S-adenosyl-l-homocysteine hydrolase. The pharmacological approach with exogenous Hcy resulted in significant rise in lipid peroxidation and more potent late blight disease development in leaves of susceptible potato as well. Finally, it has been found that key defense enzymes, i.e. phenylalanine ammonia lyase and β-1,3-glucanase were up-regulated early in the resistant potato genotype, starting from 1st hpi. In turn, in the susceptible potato the time-lag in expression of these enzymes tuned with excess production of Hcy might facilitate leaf tissue colonization by pathogen. Based on obtained results it should be stated that Hcy over-accumulation is engaged in pathophysiological mechanism leading to the abolishment of the resistance and might be an informative disease hallmark both in plant and in animal system.

We investigated how potato exposed to a chemical agent could activate nitric oxide (NO)-dependent events facilitating more potent defense responses to a subsequent pathogen attack. Obtained data revealed that all applied inducers, i.e., β-aminobutyric acid (BABA), γ-aminobutyric acid (GABA), laminarin, or 2,6-dichloroisonicotinic acid (INA), were active stimuli in potentiating NO synthesis in the primed potato. It is assumed, for the mechanism proposed in this paper, that priming involves reversible S-nitrosylated protein (S-nitrosothiols [SNO]) storage as one of the short-term stress imprint components, apart from epigenetic changes sensitized by NO. Based on BABA- and GABA-induced events, it should be stated that a rise in NO generation and coding the NO message in SNO storage at a relatively low threshold together with histone H2B upregulation might create short-term imprint activation, facilitating acquisition of a competence to react faster after challenge inoculation. Laminarin elicited strong NO upregulation with an enhanced SNO pool–altered biochemical imprint in the form of less effective local recall, nevertheless being fully protective in distal responses against P. infestans. In turn, INA showed the most intensified NO generation and abundant formation of SNO, both after the inducer treatment and challenge inoculation abolishing potato resistance against the pathogen. Our results indicate, for the first time, that a precise control of synthesized NO in cooperation with reversible SNO storage and epigenetic modifications might play an important role in integrating and coordinating defense potato responses in the priming phenomenon.

The primed mobilization for more potent defense responses to subsequent stress has been shown for many plant species, but there is a growing need to identify reliable molecular markers for this unique phenomenon. In the present study a proteomic approach was used to screen similarities in protein abundance in leaves of primed potato (Solanum tuberosum L.) treated with four well-known inducers of plant resistance, i.e. β-aminobutyric acid (BABA), γ-aminobutyric acid (GABA), Laminarin and 2,6-dichloroisonicotinic acid (INA), respectively. Moreover, to gain insight into the importance of nitric oxide (NO) in primed protein accumulation the potato leaves were supplied by S-nitrosoglutathione (GSNO), as an NO donor. The comparative analysis, using two-dimensional electrophoresis and mass spectrometry, revealed that among 25 proteins accumulated specifically after BABA, GABA, INA and Laminarin treatments, 13 proteins were accumulated also in response to GSNO. Additionally, overlapping proteomic changes between BABA-primed and GSNO-treated leaves showed 5 protein spots absent in the proteome maps obtained in response to the other priming agents. The identified 18 proteins belonged, in most cases, to functional categories of primary metabolism. The selected proteins including three redox-regulated enzymes, i.e. glyceraldehyde 3-phosphate dehydrogenase, carbonic anhydrase, and fructose-bisphosphate aldolase, were discussed in relation to the plant defence responses. Taken together, the overlapping effects in the protein profiles obtained between priming agents, GSNO and cPTIO treatments provide insight indicating that the primed potato exhibits unique changes in the primary metabolism, associated with selective protein modification via NO.

The sequence of events leading to the programmed cell death (PCD) induced by heavy metals in plants is still the object of extensive investigation. In this study we showed that roots of 3-day old yellow lupine (Lupinus luteus L.) seedlings exposed to cadmium (Cd, 89μM CdCl(2)) resulted in PCD starting from 24h of stress duration, which was evidenced by TUNEL-positive reaction. Cd-induced PCD was preceded by a relatively early burst of nitric oxide (NO) localized mainly in the root tips. Above changes were accompanied by the NADPH-oxidase-dependent superoxide anion (O(2)(·-)) production. However, the concomitant high level of both NO and O(2)(·-) at the 24th h of Cd exposure did not provoke an enhanced peroxynitrite formation. The treatment with the NADPH-oxidase inhibitor and NO-scavenger significantly reduced O(2)(·-) and NO production, respectively, as well as diminished the pool of cells undergoing PCD. The obtained data indicate that boosted NO and O(2)(·-) production is required for Cd-induced PCD in lupine roots. Moreover, we found that in roots of 14-day old lupine plants the NO-dependent Cd-induced PCD was correlated with the enhanced level of the post-stress signals in leaves, including distal NO cross-talk with hydrogen peroxide.

During the last decade it has been found that cadmium (Cd), one of the most toxic elements occurring in polluted environments, interferes with nitric oxide (NO), a multifunctional signaling molecule in living organisms. The formation of NO has been demonstrated in vivo in various plant tissues exposed to Cd stress, but unfortunately, the time and intensity of NO generation, relatively frequently shows conflicting data. What is more, there is still limited information regarding the functional role of endogenously produced NO in plants challenged with heavy metals. The first pharmacological approaches revealed that exogenously applied NO can alleviate cadmium toxicity in plants, promoting the direct scavenging of reactive oxygen species (ROS) or activating antioxidant enzymes. However, recent reports have indicated that NO even contributes to Cd toxicity by promoting Cd uptake and participates in metal-induced reduction of root growth. In view of this heterogeneous knowledge, much more puzzling if we consider results first obtained using exogenous NO sources, this review is focused mainly on the implication of endogenous NO in plant response to Cd exposure. Furthermore, a basic draft for NO mode of action during cadmium stress is proposed.

Summary Changes in electrolyte leakage, and Ca +2 , K + , and CI - effluxes from potato ( Solanum tuberosum L.) tubers in response to either cadmium or Phytophthora infestans , or both, were investigated. Two cultivars, Bintje and Bzura, susceptible and resistant, respectively, to pathogen and cadmium were examined. Longterm exposure of the plant to cadmium, resulting in the presence of cadmium in tubers (0.12–20μgg -1 fw) caused an increase of membrane integrity in the organs of both cultivars. As a result of the interaction between the tuber slices and pathogen marked increases in electrolyte leakage, and in K + , and C 1- effluxes from susceptible material as well as in Ca +2 efflux from resistant material were observed. A simultaneous treatment of potato tubers of both cultivars with cadmium and pathogen brought about antagonistic effects in membrane permeability.

Summary Changes of electrolyte leakage from potato Solanum tuberosum L. leaves effected either by cadmium, or Phytophthora infestans , or both simultaneously were investigated. The leaves of two cultivars, Bintje and Bzura, susceptible and resistant, respectively, to pathogen were examined. Cadmium present in the leaves (8–19 µg g −1 fw) caused an increase of membrane permeability in the resistant leaves and a significant decrease in the susceptible material. The interaction between the leaves and pathogen produced a marked increase in electrolyte leakage from the susceptible material and a slight increase from the resistant material. The simultaneous treatment of potato leaves of both cultivars with cadmium and the pathogen brought about antagonistic effects in membrane permeability.

Participation of nitric oxide (NO) in cross-talk between ivy pelargonium (Pelargonium peltatum) leaves and Botrytis cinerea was investigated using electrochemical and biochemical approaches. In response to the necrotroph, leaves initiated a near-immediate NO burst, but the specificity of its generation was dependent on the genetic makeup of the host plant. In the resistant cultivar, a strong NO burst was followed by a wave of secondary NO generation, shown by bio-imaging with DAF-2DA. The epicentre of NO synthesis was located in targeted cells, which exhibited a TUNEL-positive reaction. Soon after the challenge, an elevated concentration of hydrogen peroxide (H(2)O(2)) was correlated with a reversible inhibition of catalase (CAT), ascorbate peroxidase (APX), and suppression of ethylene synthesis. The induced NO generation initially expanded and then gradually disappeared on successive days, provoking noncell-death-associated resistance with an enhanced pool of antioxidants, which finally favoured the maintenance of homeostasis of surrounding cells. By contrast, in the susceptible pelargonium, a weak NO burst was recorded and further NO generation increased only as the disease progressed, which was accompanied by very intensive H(2)O(2) and ethylene synthesis. The pathogen colonizing susceptible cells also acquired the ability to produce considerable amounts of NO and enhanced nitrosative and oxidative stress in host tissues.

The paper deals with photometric and morphologic analysis of bioluminescent images obtained by registration of light radiated directly from some plant objects. Registration of images obtained from ultra-weak light sources by the single photon counting (SPC) technique is the subject of this work. The radiation is registered by use of a 16-bit charge coupled device (CCD) camera "Night Owl" together with WinLight EG&G Berthold software. Additional application-specific software has been developed in order to deal with objects that are changing during the exposition time. Advantages of the elaborated set of easy configurable tools named FCT for a computer-aided photometric and morphologic analysis of numerous series of quantitatively imperfect chemiluminescent images are described. Instructions are given how to use these tools and exemplified with several algorithms for the transformation of images library. Using the proposed FCT set, automatic photometric and morphologic analysis of the information hidden within series of chemiluminescent images reflecting defensive processes in poinsettia (Euphorbia pulcherrima Willd) leaves affected by a pathogenic fungus Botrytis cinerea is revealed.