L-Aminoguanidine Induces Imbalance of ROS/RNS Homeostasis and Polyamine Catabolism of Tomato Roots after Short-Term Salt Exposure.

Polyamine (PA) catabolism mediated by amine oxidases is an important process involved in fine-tuning PA homeostasis and related mechanisms during salt stress. The significance of these amine oxidases in short-term responses to salt stress is, however, not well understood. In the present study, the effects of L-aminoguanidine (AG) on tomato roots treated with short-term salt stress induced by NaCl were studied. AG is usually used as a copper amine oxidase (CuAO or DAO) inhibitor. In our study, other alterations of PA catabolism, such as reduced polyamine oxidase (PAO), were also observed in AG-treated plants. Salt stress led to an increase in the reactive oxygen and nitrogen species in tomato root apices, evidenced by in situ fluorescent staining and an increase in free PA levels. Such alterations were alleviated by AG treatment, showing the possible antioxidant effect of AG in tomato roots exposed to salt stress. PA catabolic enzyme activities decreased, while the imbalance of hydrogen peroxide (H2O2), nitric oxide (NO), and hydrogen sulfide (H2S) concentrations displayed a dependence on stress intensity. These changes suggest that AG-mediated inhibition could dramatically rearrange PA catabolism and related reactive species backgrounds, especially the NO-related mechanisms. More studies are, however, needed to decipher the precise mode of action of AG in plants exposed to stress treatments.

Effect of ascorbate and hydrogen peroxide on hormone and metabolite levels during post-germination growth in wheat.

The modulation of hormone and metabolite levels by ascorbate (ASA) and hydrogen peroxide (H2 O2 ) was compared during post-germination growth in shoots of wheat. Treatment with ASA resulted in a greater reduction of growth than the addition of H2 O2 . ASA also had a larger effect on the redox state of the shoot tissues as shown by the higher ASA and glutathione (GSH) levels, lower glutathione disulfide (GSSG) content and GSSG/GSH ratio compared to the H2 O2 treatment. Apart from common responses (i.e., increase of cis-zeatin and its O-glucosides), the contents of several compounds related to cytokinin (CK) and abscisic acid (ABA) metabolism were greater after ASA application. These differences in the redox state and hormone metabolism following the two treatments may be responsible for their distinct influence on various metabolic pathways. Namely, the glycolysis and citrate cycle were inhibited by ASA and they were not affected by H2 O2 , while the amino acid metabolism was induced by ASA and repressed by H2 O2 based on the changes in the level of the related carbohydrates, organic and amino acids. The first two pathways produce reducing power, while the last one needs it; therefore ASA, as a reductant may suppress and induce them, respectively. H2 O2 as an oxidant had different effect, namely it did not alter glycolysis and citrate cycle, and inhibited the formation of amino acids.

Metabolite profiling of tubers of an early- and a late-maturing potato line and their grafts.

INTRODUCTION: Earliness of tuberisation and the quality of potato tubers are important traits in potato breeding. The qualitative traits rely on the metabolite profile of tubers, which are storage organs and net importers of assimilates. Thus, the quality of tubers largely depends on the metabolites transported from leaves to developing tubers. OBJECTIVES: To test the influence of canopy on the quality of tubers by metabolite profiling of tubers of an early- and a late-maturing potato line and their grafts. METHODS: Potatoes were grown under greenhouse conditions, grafted and the tubers harvested at the end of the scions' vegetation period. Metabolite profiling of freshly harvested tubers was performed using gas chromatography coupled with mass spectrometry. Statistical analyses were applied to determine the significant differences between the different tubers. RESULTS: 99 metabolites were identified and an additional 181 peaks detected in chromatograms, out of which 186 were polar and 94 non-polar compounds. The concentrations of 113 metabolites were significantly different in the tubers from the early-maturing CE3130 and the late-maturing CE3027 line. Hetero-grafting resulted in considerable changes in the metabolite content of tubers. Especially, the effect of CE3027 on the metabolite composition of tubers formed on CE3130 rootstocks was readily apparent. Nevertheless, many compounds were present at similar levels in the tubers of hetero-grafted plants as was found in the tubers of their scion counterparts. CONCLUSION: Hetero-grafting resulted in many compounds at similar concentrations in rootstock tubers as in scion tubers suggesting that these are transported from the source leaves to tubers.

The role of methyl salicylate in plant growth under stress conditions.

Methyl salicylate is a volatile compound, the synthesis of which takes place via the salicylic acid pathway in plants. Both compounds can be involved in the development of systemic acquired resistance and they play their role partly independently. Salicylic acid transport has an important role in long-distance signalling, but methyl salicylate has also been suggested as a phloem-based mobile signal, which can be demethylated to form salicylic acid, inducing the de-novo synthesis of salicylic acid in distal tissue. Despite the fact that salicylic acid has a protective role in abiotic stress responses and tolerance, very few investigations have been reported on the similar effects of methyl salicylate. In addition, as salicylic acid and methyl salicylate are often treated simply as the volatile and non-volatile forms of the same compound, and in several cases they also act in the same way, it is hard to highlight the differences in their mode of action. The main aim of the present review is to reveal the individual role and action mechanism of methyl salicylate in systemic acquired resistance, plant-plant communication and various stress conditions in fruits and plants.

Light Spectral Composition Modifies Polyamine Metabolism in Young Wheat Plants.

Although light-emitting diode (LED) technology has extended the research on targeted photomorphogenic, physiological, and biochemical responses in plants, there is not enough direct information about how light affects polyamine metabolism. In this study, the effect of three spectral compositions (referred to by their most typical characteristic: blue, red, and the combination of blue and red [pink] lights) on polyamine metabolism was compared to those obtained under white light conditions at the same light intensity. Although light quality induced pronounced differences in plant morphology, pigment contents, and the expression of polyamine metabolism-related genes, endogenous polyamine levels did not differ substantially. When exogenous polyamines were applied, their roborative effect were detected under all light conditions, but these beneficial changes were correlated with an increase in polyamine content and polyamine metabolism-related gene expression only under blue light. The effect of the polyamines on leaf gene expression under red light was the opposite, with a decreasing tendency. Results suggest that light quality may optimize plant growth through the adjustment of polyamine metabolism at the gene expression level. Polyamine treatments induced different strategies in fine-tuning of polyamine metabolism, which were induced for optimal plant growth and development under different spectral compositions.

Improvement of Drought Tolerance by Exogenous Spermidine in Germinating Wheat (Triticum aestivum L.) Plants Is Accompanied with Changes in Metabolite Composition.

Drought is one of the most important environmental factors reducing the yield and production of crops, including wheat. Polyamines are closely associated with plant stress tolerance. The present study investigated the mechanisms through seed germination with spermidine protecting wheat varieties from drought stress. In the first experiment, the effects of spermidine on the germination of wheat varieties, namely Rakhshan, Mihan, Sirvan and Pishgam, were investigated in three drought levels, namely 0, -2, and -4 MPa induced by polyethylene glycol 6000. Analysis of variance indicated that spermidine, drought stress and interaction between varieties and drought stress were significant for all traits, and with severity of stress, all traits significantly decreased. In the second experiment, detailed gene expression and non-targeted metabolomics analyses were carried out using the Rakhshan and Mihan varieties after germination, with or without spermidine treatment and/or drought stress. According to the biomass parameters, the Mihan variety showed relatively better growth compared to the other variety, but the Rakhshan one showed more pronounced responses at gene expression level to exogenous spermidine than the Mihan variety. Overall, these results showed that spermidine increased the drought tolerance of wheat at the germination stage, due to specific role of polyamine metabolism in the development of effective responses under drought stress.

Mutations in Rht-B1 Locus May Negatively Affect Frost Tolerance in Bread Wheat.

The wheat semi-dwarfing genes Rht (Reduced height) are widely distributed among the contemporary wheat varieties. These genes also exert pleiotropic effects on plant tolerance towards various abiotic stressors. In this work, frost tolerance was studied in three near-isogenic lines of the facultative variety 'April Bearded' (AB), carrying the wild type allele Rht-B1a (tall phenotype), and the mutant alleles Rht-B1b (semi-dwarf) and Rht-B1c (dwarf), and was further compared with the tolerance of a typical winter type variety, 'Mv Beres'. The level of freezing tolerance was decreasing in the order 'Mv Beres' > AB Rht-B1a > AB Rht-B1b > AB Rht-B1c. To explain the observed differences, cold acclimation-related processes were studied: the expression of six cold-related genes, the phenylpropanoid pathway, carbohydrates, amino acids, polyamines and compounds in the tricarboxylic acid cycle. To achieve this, a comprehensive approach was applied, involving targeted analyses and untargeted metabolomics screening with the help of gas chromatography/liquid chromatography­mass spectrometry setups. Several cold-related processes exhibited similar changes in these genotypes; indeed, the accumulation of eight putrescine and agmatine derivatives, 17 flavones and numerous oligosaccharides (max. degree of polymerization 18) was associated with the level of freezing tolerance in the 'April Bearded' lines. In summary, the mutant Rht alleles may further decrease the generally low frost tolerance of the Rht-B1a, and, based on the metabolomics study, the mechanisms of frost tolerance may differ for a typical winter variety and a facultative variety. Present results point to the complex nature of frost resistance.

Polyamine Metabolism under Different Light Regimes in Wheat.

Although the relationship between polyamines and photosynthesis has been investigated at several levels, the main aim of this experiment was to test light-intensity-dependent influence of polyamine metabolism with or without exogenous polyamines. First, the effect of the duration of the daily illumination, then the effects of different light intensities (50, 250, and 500 µmol m-2 s-1) on the polyamine metabolism at metabolite and gene expression levels were investigated. In the second experiment, polyamine treatments, namely putrescine, spermidine and spermine, were also applied. The different light quantities induced different changes in the polyamine metabolism. In the leaves, light distinctly induced the putrescine level and reduced the 1,3-diaminopropane content. Leaves and roots responded differently to the polyamine treatments. Polyamines improved photosynthesis under lower light conditions. Exogenous polyamine treatments influenced the polyamine metabolism differently under individual light regimes. The fine-tuning of the synthesis, back-conversion and terminal catabolism could be responsible for the observed different polyamine metabolism-modulating strategies, leading to successful adaptation to different light conditions.

Changes in physiology, gene expression and ethylene biosynthesis in MDMV-infected sweet corn primed by small RNA pre-treatment.

The physiological condition of plants is significantly affected by viral infections. Viral proliferation occurs at the expense of the energy and protein stores in infected plant cells. At the same time, plants invest much of their remaining resources in the fight against infection, making them even less capable of normal growth processes. Thus, the slowdown in the development and growth processes of plants leads to a large-scale decrease in plant biomass and yields, which may be a perceptible problem even at the level of the national economy. One form of protection against viral infections is treatment with small interfering RNA (siRNA) molecules, which can directly reduce the amount of virus that multiplies in plant cells by enhancing the process of highly conserved RNA interference in plants. The present work demonstrated how pre-treatment with siRNA may provide protection against MDMV (Maize dwarf mosaic virus) infection in sweet corn (Zea mays cv. saccharata var. Honey Koern). In addition to monitoring the physiological condition of the maize plants, the accumulation of the virus in young leaves was examined, parallel, with changes in the plant RNA interference system and the ethylene (ET) biosynthetic pathway. The siRNA pre-treatment activated the plant antiviral defence system, thus significantly reducing viral RNA and coat protein levels in the youngest leaves of the plants. The lower initial amount of virus meant a weaker stress load, which allowed the plants to devote more energy to their growth and development. In contrast, small RNA pre-treatment did not initially have a significant effect on the ET biosynthetic pathway, but later a significant decrease was observed both in the level of transcription of genes responsible for ET production and, in the amount of ACC (1-aminocyclopropane-1-carboxylic acid) metabolite. The significantly better physiological condition, enhanced RNAi response and lower quantity of virus particles in siRNA pretreated plants, suggested that siRNA pre-treatment stimulated the antiviral defence mechanisms in MDMV infected plants. In addition, the consistently lower ACC content of the plants pre-treated with siRNA suggest that ET does not significantly contribute to the successful defence in this maize hybrid type against MDMV.

Unfinished story of polyamines: Role of conjugation, transport and light-related regulation in the polyamine metabolism in plants.

Polyamines play a fundamental role in the functioning of all cells. Their regulatory role in plant development, their function under stress conditions, and their metabolism have been well documented as regards both synthesis and catabolism in an increasing number of plant species. However, the majority of these studies concentrate on the levels of the most abundant polyamines, sometimes providing data on the enzyme activity or gene expression levels during polyamine synthesis, but generally making no mention of the fact that changes in the polyamine pool are very dynamic, and that other processes are also involved in the regulation of actual polyamine levels. Differences in the distribution of individual polyamines and their conjugation with other compounds were described some time ago, but these have been given little attention. In addition, the role of polyamine transporters in plants is only now being recognised. The present review highlights the importance of conjugated polyamines and also points out that investigations should not only deal with the polyamine metabolism itself, but should also cover other important questions, such as the relationship between light perception and the polyamine metabolism, or the involvement of polyamines in the circadian rhythm.

Salicylic Acid and Sodium Salicylate Alleviate Cadmium Toxicity to Different Extents in Maize (Zea mays L.).

The role of salicylic acid in Cd tolerance has attracted more attention recently but no information is available on the efficiency of different forms of salicylic acid. The aim was thus to investigate whether both the acid and salt forms of salicylic acid provide protection against Cd stress and to compare their mode of action. Young maize plants were grown under controlled environmental conditions. One group of 10-day-old seedlings were treated with 0.5 mM SA or NaSA for 1 day then half of the pants were treated with 0.5 mM Cd for 1 day. Another group of seedlings was treated with 0.5 mM CdSO4 for 1 day without pre-treatment with SA or NaSA, while a third group was treated simultaneously with Cd and either SA or NaSA. Both salicylic acid forms reduced the Cd accumulation in the roots. Treatment with the acidic form meliorated the Cd accumulation in the leaves, while Na-salicylate increased the phytochelatin level in the roots and the amount of salicylic acid in the leaves. Furthermore, increased antioxidant enzyme activity was mainly induced by the acid form, while glutathione-related redox changes were influenced mostly by the salt form. The acidic and salt forms of salicylic acid affected the two antioxidant systems in different ways, and the influence of these two forms on the distribution and detoxification of Cd also differed. The present results also draw attention to the fact that generalisations about the stress protective mechanisms induced by salicylic acid are misleading since different forms of SA may exert different effects on the plants via separate mechanisms.

Impact of UV-B on drought- or cadmium-induced changes in the fatty acid composition of membrane lipid fractions in wheat.

UV-B radiation may have either a positive or negative impact under the same conditions in wheat, depending on the type of secondary abiotic stressor: Cd or drought. Supplemental UV-B prevented the wilting and leaf rolling induced by PEG treatment. In contrast, combined UV-B and Cd treatment resulted in pronounced oxidative stress. The opposite effect of UV-B radiation in the case of drought or cadmium stress may be related to the alteration induced in the fatty acid composition. UV-B caused changes in the unsaturation of leaf phosphatidylglycerol fractions, and the accumulation of flavonoid in the leaves may prevent the stress induced by subsequent drought treatment. However it resulted in pronounced injury despite the increased flavonoid content in roots exposed to Cd. This was manifested in a drastic decrease in the unsaturation of the leaf monogalactosyldiacylglycerol and the root phosphatidylglycerol and digalactosyldiacylglycerol fractions. Data on the flavonoid content and fatty acid composition showed that oxidative stress was induced by drought in the leaves, by Cd in the roots, and interestingly, by UV-B radiation in both the leaves and roots. The additive effect of the combined stresses was also detected in the roots. The results presented here suggest a relationship between the capacity of the plant to remodel the fatty acid composition and its resistance to various stress factors.