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Describes the beneficial effects of culture in the presence of analogue acetylsalicylic acid on the survival of potato microplant tissues subjected to the types of heat treatments used in thermotherapy. Thermotolerance of microplant tissue; Catalase activity and H[sub 2]O[sub 2] content; Implications on disease resistance and virus eradication.

The necrogenic bacterium Erwinia amylovora responsible for the fire blight disease causes cell death in apple tissues to enrich intercellular spaces with nutrients. Apple leaves contain large amounts of dihydrochalcones (DHCs), including phloridzin and its aglycone phloretin. Previous work showed an important decrease in the constitutive DHCs stock in infected leaves, probably caused by transformation reactions during the infection process. At least two flavonoid transformation pathways have been described so far: deglucosylation and oxidation. The aim of the present study was to determine whether DHCs are differentially converted in two apple genotypes displaying contrasted susceptibilities to the disease. Different analyses were performed: i) enzymatic activity assays in infected leaves, ii) identification/quantification of end-products obtained after in vitro enzymatic reactions with DHCs, iii) evaluation of the bactericidal activity of end-products. The results of the enzymatic assays showed that deglucosylation was dominant over oxidation in the susceptible genotype MM106 while the opposite was observed in the resistant genotype Evereste. These data were confirmed by LC-UV/Vis-MS analysis of in vitro reaction mixtures, especially because higher levels of o-quinoid oxidation products of phloretin were measured by using the enzymatic extracts of Evereste infected leaves. Their presence correlated well with a strong bactericidal activity of the reaction mixtures. Thus, our results suggest that a differential transformation of DHCs occur in apple genotypes with a potential involvement in the establishment of the susceptibility or the resistance to fire blight, through the release of glucose or of highly bactericidal compounds respectively., (Copyright © 2013 Elsevier Masson SAS. All rights reserved.)

Flavonoids, like other metabolites synthesized via the phenylpropanoid pathway, possess a wide range of biological activities including functions in plant development and its interaction with the environment. Dihydrochalcones (mainly phloridzin, sieboldin, trilobatin, phloretin) represent the major flavonoid subgroup in apple green tissues. Although this class of phenolic compounds is found in very large amounts in some tissues (≈200mg/g of leaf DW), their physiological significance remains unclear. In the present study, we highlight their tissue-specific localization in young growing shoots suggesting a specific role in important physiological processes, most notably in response to biotic stress. Indeed, dihydrochalcones could constitute a basal defense, in particular phloretin which exhibits a strong broad-range bactericidal and fungicidal activity. Our results also indicate that sieboldin forms complexes with iron with strong affinity, reinforcing its antioxidant properties and conferring to this dihydrochalcone a potential for iron seclusion and/or storage. The importance of localization and biochemical properties of dihydrochalcones are discussed in view of the apple tree defense strategy against both biotic and abiotic stresses., (Copyright © 2013 Elsevier Ltd. All rights reserved.)

Fire blight is a bacterial disease of Maloideae caused by Erwinia amylovora (Ea). This necrogenic enterobacterium uses a type III secretion system (T3SS) to inject type III effectors into the plant cells to cause disease on its susceptible hosts, including economically important crops like apple and pear. The expressions of marker genes of the salicylic acid (SA) and jasmonic acid (JA) defense regulation pathways were monitored by RT-qPCR in leaves of two apple genotypes, one susceptible and one resistant, challenged with a wild type strain, a T3SS-deficient strain or water. The transcriptional data taken together with hormone level measurements indicated that the SA pathway was similarly induced in both apple genotypes during infection by Ea. On the contrary, the data clearly showed a strong T3SS-dependent down-regulation of the JA pathway in leaves of the susceptible genotype but not in those of the resistant one. Accordingly, methyl-jasmonate treated susceptible plants displayed an increased resistance to Ea. Bacterial mutant analysis indicated that JA manipulation by Ea mainly relies on the type III effector DspA/E. Taken together, our data suggest that the T3SS-dependent down-regulation of the JA pathway is a critical step in the infection process of Malus spp. by Ea., (Copyright © 2012 Elsevier Ireland Ltd. All rights reserved.)

Climate change is projected to have a significant ecological impact on natural ecosystems, most notably through direct and indirect modifications of local precipitation regimes. In addition, anthropic activities such as the removal of vegetation, soil proofing due to building, the absence of storm drains and crop over-irrigation will all increase the occurrence of flooding. As a result, forest species, and more specifically trees, will increasingly be exposed to soil waterlogging. It is now well established that such flooding events can lead to changes in forest distribution and composition. For such reasons, it is becoming increasingly important to study forest ecosystems and more particularly the adaptive potential of tree species to better understand the ecological plasticity of forest communities to environmental modifications.

Soil flooding is an environmental constraint that is increasingly important for forest ecosystems, affecting tree growth and regeneration. As a result, selection pressure will alter forest diversity and distribution by favouring tree species tolerant of soil oxygen deprivation. Sessile and pedunculate oaks are the most abundant oak species and they exhibit a strong differential tolerance to waterlogging. In order to gain some understanding of the mechanisms of tolerance of both species to hypoxia, we undertook the characterization of the physiological, morphological, cellular and molecular responses of both species to flooding stress. Our results indicate that pedunculate oak, the more tolerant species, succeeded in maintaining its growth, water status and photosynthetic activity at a higher level than sessile oak. Furthermore, pedunculate oak developed aerenchyma in its root cortex as well as adventitious roots. The later exhibited a strong accumulation of class1 non-symbiotic haemoglobin localized by in situ hybridization in the protoderm and in some cortical cells. In conclusion, the higher tolerance of pedunculate oak to flooding was associated with an enhanced capacity to maintain photosynthesis and water homeostasis, coupled with the development of adaptive features (aerenchyma, adventitious roots) and with a higher expression of non-symbiotic haemoglobin in the roots., (© 2011 Blackwell Publishing Ltd.)

Flavonoids are a group of polyphenol compounds with known antioxidant activities. Among them, dihydrochalcones are mainly found in apple leaves (Malus domestica). Glycosylated dihydrochalcones were previously found in large amounts in leaves of two genotypes of Malus with contrasting resistance to fire blight, a bacterial disease caused by Erwinia amylovora. In the present study we demonstrate that soluble polyphenol patterns comprised phloridzin alone or in combination with two additional dihydrochalcones, identified as sieboldin and trilobatin. Presence of sieboldin in young leaves correlated well with a high 1,1-diphenyl-2-picrylhydrazyl (DPPH) free radical scavenging activity. Moreover, these leaves displayed enhanced tolerance to paraquat, a photooxidative-stress generating herbicide. Interestingly, phloridzin had a high activity in the oxygen radical absorbance capacity (ORAC) assay, but its presence alone in leaves did not correlate with tolerance to paraquat. In order to further characterise the activity of these compounds, we tested their ability to prevent oxidative-dependent formation of advanced glycation end-products (AGEs) and phenylephrine-induced contraction of isolated rat mesenteric arteries. The antioxidant capacity of sieboldin was clearly demonstrated by showing that this compound (i) prevented vasoconstriction and (ii) inhibited AGEs formation. Both assays provided interesting information concerning a potential use of sieboldin as a therapeutic. Hence, our results strongly argue for a bioactivity of dihydrochalcones as functional antioxidants in the resistance of Malus leaves to oxidative stress. In addition, we demonstrate for the first time that sieboldin is a powerful multipotent antioxidant, effective in preventing physiopathological processes. Further work should aim at demonstrating the potential use of this compound as a therapeutic in treating free radical-involving diseases., (Copyright 2009 Elsevier Ltd. All rights reserved.)

The cellular and molecular adaptations of non-model woody species to environmental changes are still poorly understood. We have cloned and characterised a novel non-symbiotic hemoglobin from oak roots (QpHb1) which exhibits a specific cellular distribution in the root. The QpHb1 gene is strongly expressed in the protoderm and the protoxylem cells in two Quercus species (Q. petraea and Q. robur) with contrasting adaptive potential to drought and flooding. The constitutive expression of QpHb1 in both oak species in specific root tissues combined with the reported presence of nitric oxide in the same tissues and its potential for protein S-nitrosylation could support a role for non-symbiotic hemoglobins in signalling changes in the root environment and/or in controlling some aspects of root development.

Sessile oak is a species of great ecological and silvicultural importance in Europe; however, increased frequency and severity of flooding of forested areas pose a threat to its regeneration. We monitored water relations, root anatomical changes and the expression of two calmodulin genes (QpCaM) in sessile oak seedlings during a 14-day flooding treatment. The response followed two characteristic sequences. The first phase, in response to between 1 h and 3 days of flooding, was characterized by a dramatic but transient decline in water relations parameters followed by a recovery towards control values with no noticeable change in root cell morphology. During the second phase, in response to 3 to 14 days of flooding, water relations parameters gradually and continuously declined and hypertrophied lenticels developed at the base of the shoot. Concurrently, root cortical cells became larger and less spherical and the root cortex more porous. These cellular changes were accompanied by a transient rise in root transcript levels of QpCaM-2. We conclude that sessile oak seedlings are capable of withstanding a 3-day period of flooding without significant morphological alterations. In contrast, exposure to flooding for more than 3 days resulted in anatomical and morphological changes in the root system. These changes are, however, insufficient to provide sessile oak with long-term tolerance to flooding.

Nitric oxide (NO) and hydrogen peroxide (H(2)O(2)) are regulatory molecules in various developmental processes and stress responses. Tobacco (Nicotiana tabacum) leaves exposed to moderate high light dramatically potentiated NO-mediated cell death in catalase-deficient (CAT1AS) but not in wild-type plants, providing genetic evidence for a partnership between NO and H(2)O(2) during the induction of programmed cell death. With this experimental model system, the specific impact on gene expression was characterized by either NO or H(2)O(2) alone or both molecules combined. By means of genome-wide cDNA-amplified fragment length polymorphism analysis, transcriptional changes were compared in high light-treated CAT1AS and wild-type leaves treated with or without the NO donor sodium nitroprusside. Differential gene expression was detected for 214 of the approximately 8,000 transcript fragments examined. For 108 fragments, sequence analysis revealed homology to genes with a role in signal transduction, defense response, hormone interplay, proteolysis, transport, and metabolism. Surprisingly, only 16 genes were specifically induced by the combined action of NO and H(2)O(2), whereas the majority were regulated by either of them alone. At least seven transcription factors were mutually up-regulated, indicating significant overlap between NO and H(2)O(2) signaling pathways. These results consolidate significant cross-talk between NO and H(2)O(2), provide new insight into the early transcriptional response of plants to increased NO and H(2)O(2) levels, and identify target genes of the combined action of NO and H(2)O(2) during the induction of plant cell death.

Flooding is a major issue for plant survival in many regions of the world. Soil inundation induces multiple plant physiological dysfunctions, leading to a decline in plant growth and survival capacity. Some of the most important effects of flooding include a reduction in water and nutrient uptake and a decrease in metabolism. Prolonged soil flooding will also ultimately lead to anoxia conditions with profound effects on plant respiratory metabolism. However, it is still unclear which signals and which sensory mechanisms are responsible for triggering the plant response. In contrast, it is now established that flooding responses are typified by enhanced ethylene production, accompanied by a signalling cascade which includes a network of hormones and other common secondary signalling molecules. In recent years, there has been significant progress in the understanding of some of the signalling pathways involved during plant stress responses. Here, we present an overview of recent hypothesises on sensing and signalling during plant flooding.

In transgenic tobacco plants with reduced catalase activity, high levels of hydrogen peroxide (H2O2) can accumulate under photorespiratory conditions. Such a perturbation in H2O2 homeostasis induced cell death in clusters of palisade parenchyma cells, primarily along the veins. Ultrastructural alterations, such as chromatin condensation and disruption of mitochondrial integrity, took place before cell death. Furthermore, enhanced transcript levels of mitochondrial defense genes accompanied these mitochondrial changes. Pharmacological data indicated that the initiation and execution of cell death require de novo protein synthesis and that the signal transduction pathway leading to cell death involved changes in ion homeostasis, (de)phosphorylation events and an oxidative burst, as observed during hypersensitive responses. This oxidase-dependent oxidative burst is essential for cell death, but it is not required for the accumulation of defense proteins, suggesting a more prominent role for the oxidative burst in abiotic stress-induced cell death.

Adequate responses to environmental changes are crucial for plant growth and survival. However, the molecular and biochemical mechanisms involved are poorly understood and the signaling networks remain elusive. The accumulation of active oxygen species (AOS) is a central theme during plant responses to both biotic and abiotic stresses. In both situations, AOS can play two divergent roles: either exacerbating damage or activating multiple defense responses, thereby acting as signal molecules. Such a dual function was first described in pathogenesis, but also recently has been demonstrated during several abiotic stress responses. To allow for these different roles, cellular levels of AOS must be tightly controlled. This control can be attained through a diverse battery of oxidant scavengers. Perturbation of this scavenging capacity can lead to dramatic imbalances of AOS concentrations, leading to a modified redox status. Here, we summarize mainly the work done on plants that are deficient in catalase activity. These plants not only revealed the importance of catalase in coping with environmental stress but also provided us with a powerful tool to investigate the (multiple) roles of H2O2 in an intact plant system.

An alternative approach to maintaining indoor air quality may be the biofiltration of air circulated within the space. A biofilter with living botanical matter as the packing medium reduced concentrations of toluene, ethylbenzene, and o-xylene concurrently present at parts per billion (volume) in indoor air. The greatest reduction in concentrations per pass was under the slowest influent air flux (0.025 m s(-1)); however, the maximum amount removed per unit time occurred under the most rapid flux (0.2 m s(-1)). There was little difference between the different compounds with removal capacities of between 1.3 and 2.4 micromol m(-3) biofilter s(-1) (between 0.5 and 0.9 g m(-3) biofilter h(-1)) depending on influent flux and temperature. Contrary to biofilters subjected to higher influent concentrations, the optimal temperatures for removal by this biofilter decreased to less than 20 degrees C at the most rapid flux for all three compounds. Microbial activity was decreased at these cooler temperatures suggesting the biofilter was not microbially limited but rather was limited by the availability of substrate. The cooler temperatures allowed greater partitioning of the VOCs into the water column which had a greater impact on removal than its reduction in microbial activity.

Heat-acclimation or salicylic acid (SA) treatments were previously shown to induce thermotolerance in mustard (Sinapis alba L.) seedlings from 1.5 to 4 h after treatment. In the present study we investigated changes in endogenous SA and antioxidants in relation to induced thermotolerance. Thirty minutes into a 1-h heat-acclimation treatment glucosylated SA had increased 5.5-fold and then declined during the next 6 h. Increases in free SA were smaller (2-fold) but significant. Changes in antioxidants showed the following similarities after either heat-acclimation or SA treatment. The reduced-to-oxidized ascorbate ratio was 5-fold lower than the controls 1 h after treatment but recovered by 2 h. The glutathione pool became slightly more oxidized from 2 h after treatment. Glutathione reductase activity was more than 50% higher during the first 2 h. Activities of dehydroascorbate reductase and monodehydroascorbate reductase decreased by at least 25% during the first 2 h but were 20% to 60% higher than the control levels after 3 to 6 h. One hour after heat acclimation ascorbate peroxidase activity was increased by 30%. Young leaves appeared to be better protected by antioxidant enzymes following heat acclimation than the cotyledons or stem. Changes in endogenous SA and antioxidants may be involved in heat acclimation.

Spraying mustard (Sinapis alba L.) seedlings with salicylic acid (SA) solutions between 10 and 500 &mgr;m significantly improved their tolerance to a subsequent heat shock at 55 degreesC for 1.5 h. The effects of SA were concentration dependent, with higher concentrations failing to induce thermotolerance. The time course of thermotolerance induced by 100 &mgr;m SA was similar to that obtained with seedlings acclimated at 45 degreesC for 1 h. We examined the hypothesis that induced thermotolerance involved H2O2. Heat shock at 55 degreesC caused a significant increase in endogenous H2O2 and reduced catalase activity. A peak in H2O2 content was observed within 5 min of either SA treatment or transfer to the 45 degreesC acclimation temperature. Between 2 and 3 h after SA treatment or heat acclimation, both H2O2 and catalase activity significantly decreased below control levels. The lowered H2O2 content and catalase activity occurred in the period of maximum thermoprotection. It is suggested that thermoprotection obtained either by spraying SA or by heat acclimation may be achieved by a common signal transduction pathway involving an early increase in H2O2.