Cell Wall Peroxidase Activity Research Articles

Transcriptome and metabolome analyses reveal molecular insights into waterlogging tolerance in Barley

Waterlogging stress is one of the major abiotic stresses affecting the productivity and quality of many crops worldwide. However, the mechanisms of waterlogging tolerance are still elusive in barley. In this study, we identify key differentially expressed genes (DEGs) and differential metabolites (DM) that mediate distinct waterlogging tolerance strategies in leaf and root of two barley varieties with contrasting waterlogging tolerance under different waterlogging treatments. Transcriptome profiling revealed that the response of roots was more distinct than that of leaves in both varieties, in which the number of downregulated genes in roots was 7.41-fold higher than that in leaves of waterlogging sensitive variety after 72 h of waterlogging stress. We also found the number of waterlogging stress-induced upregulated DEGs in the waterlogging tolerant variety was higher than that of the waterlogging sensitive variety in both leaves and roots in 1 h and 72 h treatment. This suggested the waterlogging tolerant variety may respond more quickly to waterlogging stress. Meanwhile, phenylpropanoid biosynthesis pathway was identified to play critical roles in waterlogging tolerant variety by improving cell wall biogenesis and peroxidase activity through DEGs such as Peroxidase (PERs) and Cinnamoyl-CoA reductases (CCRs) to improve resistance to waterlogging. Based on metabolomic and transcriptomic analysis, we found the waterlogging tolerant variety can better alleviate the energy deficiency via higher sugar content, reduced lactate accumulation, and improved ethanol fermentation activity compared to the waterlogging sensitive variety. In summary, our results provide waterlogging tolerance strategies in barley to guide the development of elite genetic resources towards waterlogging-tolerant crop varieties.

Comparative secretome analysis of Striga and Cuscuta species identifies candidate virulence factors for two evolutionarily independent parasitic plant lineages

BackgroundMany parasitic plants of the genera Striga and Cuscuta inflict huge agricultural damage worldwide. To form and maintain a connection with a host plant, parasitic plants deploy virulence factors (VFs) that interact with host biology. They possess a secretome that represents the complement of proteins secreted from cells and like other plant parasites such as fungi, bacteria or nematodes, some secreted proteins represent VFs crucial to successful host colonisation. Understanding the genome-wide complement of putative secreted proteins from parasitic plants, and their expression during host invasion, will advance understanding of virulence mechanisms used by parasitic plants to suppress/evade host immune responses and to establish and maintain a parasite-host interaction.ResultsWe conducted a comparative analysis of the secretomes of root (Striga spp.) and shoot (Cuscuta spp.) parasitic plants, to enable prediction of candidate VFs. Using orthogroup clustering and protein domain analyses we identified gene families/functional annotations common to both Striga and Cuscuta species that were not present in their closest non-parasitic relatives (e.g. strictosidine synthase like enzymes), or specific to either the Striga or Cuscuta secretomes. For example, Striga secretomes were strongly associated with ‘PAR1’ protein domains. These were rare in the Cuscuta secretomes but an abundance of ‘GMC oxidoreductase’ domains were found, that were not present in the Striga secretomes. We then conducted transcriptional profiling of genes encoding putatively secreted proteins for the most agriculturally damaging root parasitic weed of cereals, S. hermonthica. A significant portion of the Striga-specific secretome set was differentially expressed during parasitism, which we probed further to identify genes following a ‘wave-like’ expression pattern peaking in the early penetration stage of infection. We identified 39 genes encoding putative VFs with functions such as cell wall modification, immune suppression, protease, kinase, or peroxidase activities, that are excellent candidates for future functional studies.ConclusionsOur study represents a comprehensive secretome analysis among parasitic plants and revealed both similarities and differences in candidate VFs between Striga and Cuscuta species. This knowledge is crucial for the development of new management strategies and delaying the evolution of virulence in parasitic weeds.

The Nitrate Transporter MtNPF6.8 Is a Master Sensor of Nitrate Signal in the Primary Root Tip of Medicago truncatula.

Nitrate is not only an essential nutrient for plants, but also a signal involved in plant development. We have previously shown in the model legume Medicago truncatula, that the nitrate signal, which restricts primary root growth, is mediated by MtNPF6.8, a nitrate transporter. Nitrate signal also induces changes in reactive oxygen species accumulation in the root tip due to changes in cell wall peroxidase (PODs) activity. Thus, it was interesting to determine the importance of the role of MtNPF6.8 in the regulation of the root growth by nitrate and identify the POD isoforms responsible for the changes in POD activity. For this purpose, we compared in M. truncatula a npf6.8 mutant and nitrate insensitive line deficient in MtNPF6.8 and the corresponding wild and sensitive genotype for their transcriptomic and proteomic responses to nitrate. Interestingly, only 13 transcripts and no protein were differently accumulated in the primary root tip of the npf6.8-3 mutant line in response to nitrate. The sensitivity of the primary root tip to nitrate appeared therefore to be strongly linked to the integrity of MtNPF6.8 which acts as a master mediator of the nitrate signal involved in the control of the root system architecture. In parallel, 7,259 and 493 genes responded, respectively, at the level of transcripts or proteins in the wild type, 196 genes being identified by both their transcript and protein. By focusing on these 196 genes, a concordance of expression was observed for most of them with 143 genes being up-regulated and 51 being down-regulated at the two gene expression levels. Their ontology analysis uncovered a high enrichment in POD genes, allowing the identification of POD candidates involved in the changes in POD activity previously observed in response to nitrate.

Elevated carbon dioxide alleviates the negative impact of drought on wheat by modulating plant metabolism and physiology

This study was conducted to understand the mechanism of wheat yield decrease under drought stress and the role of CO2 in modulating physiological and metabolic drought effects. Wheat was grown under ambient and elevated CO2 (400 and 800 ppm, respectively), and plants were subjected to drought stress prior to anthesis. Photosynthetic rate (An), stomatal conductance (Gs), transpiration rate (E) and activities of carbohydrate metabolic enzymes were decreased in leaf and increased in spikes during drought. Total antioxidant potential (TAP) was decreased under drought both in leaf and spike. Grain yield parameters were again reduced under drought, while An, E and most of the yield traits were increased under elevated CO2. The number of grains spike-1 correlated positively with An, TAP and cell wall invertase activity, while it negatively correlated with ascorbate peroxidase, cell wall peroxidase and glutathione reductase activities in leaves. Thousand kernel weight positively correlated with leaf phosphoglucoisomerase and spike glucose-6-phosphate dehydrogenase activities. This indicates that elevated CO2 could boost CO2 assimilation through an increase in antioxidant potential and facilitate more photosynthate supply via various increased carbohydrate metabolic enzyme activities, and thus increases yield. This could be a possible mechanism of grain yield increase caused by elevated CO2.

A biochemical and proteomic approach to the analysis of tomato mutant fruit growth

To assess the effects of ABA deficiency on tomato fruit growth, the ABA mutant flacca was grown in an optimal soil water regime and various analyzes were performed, including morphological (fruit number, diameter and fruit biomass), physiological (duration of growth and fruit growth rate), biochemical (ABA accumulation, enzyme cell wall peroxidase activity) as well as proteomics. The fruit growth analysis showed that the slower fruit growth rate and development resulted in smaller flacca fruits in comparison to the wild-type fruits. The comparison of the temporal dynamics of cell wall peroxidase activity and ABA content in our experiment indicated an opposite relationship during fruit development. Proteomic analysis and the down-regulation of most proteins from carbon and amino acid metabolism, the translation and processing of proteins, energy metabolism and cell wall-related metabolism in the flacca fruits compared to the wild type, indicated reduced metabolic flux which reflected a slower fruit growth and development and reduced fruit size in the ABA mutant. These findings also indicated that ABA limited carbon sources, which could be responsible for the reduced fruit growth and size of ABA-deficient tomato fruits. The up-regulation of sulfur and oxygen-evolving enhancer proteins in the flacca fruits implicated the maintenance of photosynthesis in the late expansion phase, which slows down transition to the ripening stage. The majority of antioxidative and stress defence proteins were down-regulated in the flacca fruits, which could be related to the role of ABA in the activity of different antioxidative enzymes as well as in regulating cell wall expansion and the cessation of fruit growth.

Arbuscular mycorrhizal fungi promote lead immobilization by increasing the polysaccharide content within pectin and inducing cell wall peroxidase activity

The mechanism by which arbuscular mycorrhizal (AM) fungi immobilize lead (Pb) within the cell wall is unclear. Therefore, the aim of this study was to investigate the mechanism by which AM fungi immobilize Pb within the cell wall by measuring the Pb content in the cell wall, the polysaccharide and the uronic acid contents of different cell wall fractions, and the activity of cell wall peroxidase. Mycorrhizal-associated Medicago truncatula had higher shoot and root biomass than nonmycorrhizal-associated M. truncatula. AM inoculation increased the content of Pb in the cell wall under Pb stress. The polysaccharide content in the pectin and hemicellulose fractions were increased by AM inoculation with or without Pb stress. In AM-associated roots, the cell wall peroxidase activity increased in response to Pb stress. However, Pb stress did not affect the cell wall peroxidase activity in non-AM-associated roots. Correlation analysis suggested that MtPrx05 and MtPrx10 may participate in polysaccharide cross-linking and cell wall stiffening. The Pb stress resistance mechanism of AM-associated roots may involve cell wall stiffening. Taken together, the results show that AM inoculation may improve host plant growth and increase Pb immobilization in the cell wall by increasing the polysaccharide content within pectin and hemicellulose and by inducing cell wall peroxidase activity. Both the polysaccharide composition and cell wall peroxidase have important contributions to the resistance of mycorrhizal-associated plants.

Activities of leaf and spike carbohydrate-metabolic and antioxidant enzymes are linked with yield performance in three spring wheat genotypes grown under well-watered and drought conditions

BackgroundTo improve our understanding about the physiological mechanism of grain yield reduction at anthesis, three spring wheat genotypes [L1 (advanced line), L2 (Vorobey) and L3 (Punjab-11)] having contrasting yield potential under drought in field were investigated under controlled greenhouse conditions, drought stress was imposed at anthesis stage by withholding irrigation until all plant available water was depleted, while well-watered control plants were kept at 95% pot water holding capacity.ResultsCompared to genotype L1 and L2, pronounced decrease in grain number (NGS), grain yield (GY) and harvest index (HI) were found in genotype L3, mainly due to its greater kernel abortion (KA) under drought. A significant positive correlation of leaf monodehydroascorbate reductase (MDHAR) with both NGS and HI was observed. In contrast, significant negative correlations of glutathione S-transferase (GST) and vacuolar invertase (vacInv) both within source and sink were found with NGS and HI. Likewise, a significant negative correlation of leaf abscisic acid (ABA) with NGS was noticed. Moreover, leaf aldolase and cell wall peroxidase (cwPOX) activities were significantly and positively associated with thousand kernel weight (TKW).ConclusionDistinct physiological markers correlating with yield traits and higher activity of leaf aldolase and cwPOX may be chosen as predictive biomarkers for higher TKW. Also, higher activity of MDHAR within the leaf can be selected as a predictive biomarker for higher NGS in wheat under drought. Whereas, lower activity of vacInv and GST both within leaf and spike can be selected as biomarkers for higher NGS and HI. The results highlighted the role of antioxidant and carbohydrate-metabolic enzymes in the modulation of source-sink balance in wheat crops, which could be used as bio-signatures for breeding and selection of drought-resilient wheat genotypes for a future drier climate.

Overexpression of the wheat expansin gene TaEXPA2 improves oxidative stress tolerance in transgenic Arabidopsis plants

Expansins play an important role in plant stress tolerance. In a previous study, we cloned the wheat expansin gene TaEXPA2. Here, we analyze its involvement in oxidative stress tolerance. First, we observed that the expression of TaEXPA2 in wheat seedlings was upregulated during H2O2 stress. Then, we assembled a TaEXPA2 gene expression vector, transformed it to Arabidopsis, and obtained transgenic plants overexpressing TaEXPA2 (labeled OE). When exposed to H2O2, both OE and wild-type (Col) plants were damaged by oxidative stress, as indicated by decolored leaves and increased malondialdehyde (MDA) content. Damage in OE plants was less severe than in Col plants (WT), and this was accompanied by higher activity of cell wall peroxidase (POD) enzymes, including soluble POD, ionically bound POD, and covalently bound POD. The expansin activities of the OE plants were also higher than WT under oxidative stress. We further obtained the Arabidopsis mutant atexpa2 (AtEXPA2 is homologous to TaEXPA2), and found that the antioxidant ability of atexpa2 was lower than that in Col plants, accompanied by depressed activity of POD enzymes and expansins in cell walls. We transformed wheat TaEXPA2 to atexpa2 and obtained plants (labeled Rs) capable of recovering the antioxidant capacity. Oxidative stress tolerance in Rs plants was higher than that of Col plants, and the Rs plants also had higher levels of cell wall POD enzyme and expansin activity. Finally, we identified 13 POD genes in Arabidopsis thaliana and analyzed their expression patterns using quantitative real-time PCR. The expression of 4 of these genes (AtPOD31, AtPOD33, AtPOD34 and AtPOD71) was significantly upregulated during exposure to H2O2. We speculate that the 4 genes upregulated by H2O2 treatment are involved in the increased activity of POD in the cell wall. We suggest that TaEXPA2 may regulate antioxidant capacity in plants by regulating the activity of cell wall peroxidase.

Ascorbate deficiency influences the leaf cell wall glycoproteome in Arabidopsis thaliana

The cell wall forms the first line of interaction between the plant and the external environment. Based on the observation that ascorbate-deficient vtc mutants of Arabidopsis thaliana have increased cell wall peroxidase activity, the cell wall glycoproteome of vtc2-2 was investigated. Glycoproteins were purified from fully expanded leaves by Concanavalin A affinity chromatography and analysed by liquid chromatography quadrupole time-of-flight mass spectrometry. This procedure identified 63 proteins with predicted glycosylation sites and cell wall localization. Of these, 11 proteins were differentially expressed between vtc2-2 and wild type. In particular, PRX33/34 were identified as contributing to increased peroxidase activity in response to ascorbate deficiency. This is the same peroxidase previously shown to contribute to hydrogen peroxide generation and pathogen resistance. Three fasciclin-like arabinogalactan proteins (FLA1, 2 and 8) had lower abundance in vtc2-2. Inspection of published microarray data shows that these also have lower gene expression in vtc1 and vtc2-1 and are decreased in expression by pathogen challenge and oxidative stresses. Ascorbate deficiency therefore impacts expression of cell wall proteins involved in pathogen responses and these presumably contribute to the increased resistance of vtc mutants to biotrophic pathogens.

Effect of Auxin on Weedy Rice Mesocotyl Cell Wall Oxidase

The seedling was developed on the dark condition by artificial climate chamber to determine the content of Auxin (IAA), activity of indoleacetic acid oxidase (IAO) and cell wall peroxidase (POD), mesocotyl length change and cell wall oxidase activity variation applied IAA. The study proves that endogenous hormone IAA content of long mesocotyl in weedy rice were much higher than that of short mesocotyl in Akimitsu. With the growth of mesocotyl elongation, endogenous IAA content showed cumulative effects. And then accumulation of IAA content reached up to maximum, when the Mesocotyl elongation stoped to grow. IAA might play a decisive role in the process of hypocotyl elongation. Reduction in the activity of IAO and POD accelerated the transformation from bound IAA to free IAA and promoted cell elongation and mesocotyl elongation.

Production of hydrogen peroxide and expression of ROS‐generating genes in peach flower petals in response to host and non‐host fungal pathogens

Reactive oxygen species (ROS) play dual roles in plant–microbe interactions in that they can either stimulate host resistance or enhance pathogen virulence. Innate resistance in peach (Prunus persica) to the brown rot fungal pathogen Monilinia fructicola is very limited, and knowledge of the mechanism of virulence is rudimentary. In this study, production of hydrogen peroxide, a major component of ROS, was determined in peach flower petals in response to M. fructicola (a host pathogen) and Penicillium digitatum (a non‐host pathogen). Monilinia fructicola was able to infect flower petals while P. digitatum was not. During the host‐specific interaction, M. fructicola induced hydrogen peroxide accumulation in flower petals. Application of exogenous antioxidants significantly reduced hydrogen peroxide accumulation as well as the incidence of brown rot disease. Application of M. fructicola spores to the surface of intact flower petals induced gene expression and increased enzyme activity of NADPH oxidase and cell wall peroxidase in host tissues, resulting in the production of hydrogen peroxide. Petals inoculated with M. fructicola exhibited high levels of protein carbonylation and lipid peroxidation. No significant response in gene expression, enzyme activity or hydrogen peroxide levels was observed in peach flower petals treated with P. digitatum. These results suggest that M. fructicola, as with other necrotrophic fungi, uses the strong oxidative response as part of a virulence mechanism.

Heme Oxygenase‐1 is Associated with Wheat Salinity Acclimation by Modulating Reactive Oxygen Species HomeostasisF

Heme oxygenase-1 (HO-1) has been recently identified as an endogenous signaling system in animals. In this study, HO-1 upregulation and its role in acquired salt tolerance (salinity acclimation) were investigated in wheat plants. We discovered that pretreatment with a low concentration of NaCl (25 mmol/L) not only led to the induction of HO-1 protein and gene expression, as well as enhanced HO activity, but also to a salinity acclimatory response thereafter. The effect is specific for HO-1, since the potent HO-1 inhibitor zinc protoporphyrin IX blocks the above cytoprotective actions, and the cytotoxic responses conferred by 200 mmol/L NaCl are reversed partially when HO-1 inducer hemin is added. Heme oxygenase catalytic product, carbon monoxide (CO) aqueous solution pretreatment, mimicked the salinity acclimatory responses. Meanwhile, the CO-triggered re-establishment of reactive oxygen species (ROS) homeostasis was mainly guaranteed by the induction of total and isozymatic activities, or corresponding transcripts of superoxide dismutase, ascorbate peroxidase, and cytosolic peroxidase (POD), as well as the downregulation of NADPH oxidase expression and cell-wall POD activity. A requirement of hydrogen peroxide homeostasis for HO-1-mediated salinity acclimation was also discovered. Taken together, the above results suggest that the upregulation of HO-1 expression was responsible for the observed salinity acclimation through the regulation of ROS homeostasis.

Beneficial effects of silicon nutrition in alleviating salinity stress in hydroponically grown canola, Brassica napus L., plants

Silicon (Si) is the second most abundant element in soil and effectively counteracts the effects of various abiotic stresses, such as drought, heavy metal toxicity and salinity, on plants. In the present study the ameliorating effects of Si nutrition supplied as 2 mmol L−1 sodium silicate were investigated on hydroponically grown canola (Brassica napus L.) plants under salinity stress (i.e. 150 mmol L−1 sodium chloride). Salinity decreased plant growth parameters such as tissue fresh and dry weights. These decreases were accompanied by increased lignin contents, Na+ ion accumulation, increased lipid peroxidation and decreased chlorophyll contents in plants. Silicon nutrition, however, enhanced plant growth parameters and led to the prevention of lignin and the Na+ accumulation in shoots, reduced levels of lipid peroxidation in the roots and higher levels of chlorophyll. As a result of salinity, catalase activity in the whole plant and both soluble and cell wall peroxidase activities in the shoots decreased. Silicon nutrition, however, increased the reactive oxygen species scavenging capacity of salt-stressed plants through increased catalase and cell wall peroxidase activities. Thus, silicon nutrition ameliorated the deleterious effects of salinity on the growth of canola plants through lower tissue Na+ contents, maintaining the membrane integrity of root cells as evidenced by reduced lipid peroxidation, increased reactive oxygen species scavenging capacity and reduced lignification.

Changes in cucumber hypocotyl cell wall dynamics caused by Azospirillum brasilense inoculation

We previously reported that Azospirillum brasilense induced a more elastic cell wall and a higher apoplastic water fraction in both wheat coleoptile and flag leaf. These biophysical characteristics could permit increased growth. Knowledge of the biochemical effects the bacteria could elicit in plant cell walls and how these responses change plant physiology is still scarce. The objective of this work was to analyze whether A. brasilense Sp245 inoculation affected elongation and extensibility of growing cucumber (Cucumis sativus) hypocotyls and ionically bound cell wall peroxidase activities. Hypocotyl tip and basal segments were excised from A. brasilense Sp245-inoculated cucumber seedlings growing in darkness under hydroponic conditions. Elongation, cell wall extensibility, cell wall peroxidase activities against ferulic acid and guaiacol and NADH oxidase activities were analyzed. Azospirillum-inoculated cucumber seedlings grew bigger than non-inoculated ones. Dynamic cell wall differences were detected between inoculated and non-inoculated hypocotyls. They included greater acid-induced cell wall extension and in vivo elongation when incubated in distilled water. Although there was no difference between treatments in either region of the hypocotyl NADH oxidase and ferulic acid peroxidase activities were lower in both regions in inoculated seedlings. These lesser activities could be delaying the stiffening of cell wall in inoculated seedlings. These results showed that the cell wall is a target for A. brasilense growth promotion.

Structural Changes of Cell Wall and Lignifying Enzymes Modulations in Bean Roots in Response to Copper Stress

Fourteen-day-old bean seedlings were cultured in nutrient solution containing Cu(2+) ions at various concentrations (50 and 75 microM of CuSO(4)) for 3 days. This excess of copper induced a reduction in the water volume absorbed by the plants. Moreover, this reduction was accompanied by an increase of the amount of copper taken up by the roots. Analysis by native gel electrophoresis of cell wall peroxidase activities in the roots revealed a stimulation of two anionic isoforms (A(2) and A(3)) under cupric stress conditions. Moreover, the activity of phenylalanine ammonia lyase (EC. 4.3.1.5), which plays an important role in plant defense, was enhanced. Copper-treated bean roots showed modifications in the cell walls of various tissues. Label for lignin, callose, and suberin with berberine-aniline blue revealed abnormal cell wall thickenings in the endodermis, the phloem, and the xylem, especially in plants treated with 75 microM CuSO(4).

Ultraviolet Radiation as a Limiting Factor in Leaf Expansion and Development

Reductions in leaf growth are a commonly observed response to ultraviolet radiation, but the underlying mechanisms remain poorly defined. This study examined the response of leaves exposed to a UV environment across a range of organizational scales, including leaf expansion rate, epidermal cell size and number, biomechanical properties, leaf-water relations and activity of cell-wall peroxidases. Two experimental approaches were used; Lettuce (Lactuca sativa L.) plants were propagated under (a) supplementary UV-B (9 kJ m(-2) day(-1)) in controlled environment (CE) conditions, and (b) field conditions, where plants were placed under three horticultural films with differing UV transmissions. In both experiments, UV-B caused the greatest reductions in leaf expansion and final leaf size, with some reductions attributable to UV-A wavelengths. In supplementary UV-B conditions, adaxial cell size was reduced, while in field plants, both cell size and cell number were lower in an increased UV environment, as was the case with abaxial cells in CE plants. Although leaf turgor and leaf extensibility were not affected by UV wavelengths, breaking strain of leaf tissue was decreased under supplementary UV-B. Cell-wall peroxidase activity was increased in both supplementary UV conditions and in the field, where only a zero UV environment showed no upregulation of cell-wall peroxidase.

Antioxidant status, peroxidase activity, and PR protein transcript levels in ascorbate-deficient Arabidopsis thaliana vtc mutants

Ascorbate is the most abundant small molecule antioxidant in plants and is proposed to function, along with other members of an antioxidant network, in controlling reactive oxygen species. A biochemical and molecular characterization of four ascorbate-deficient (vtc) Arabidopsis thaliana mutants has been carried out to determine if ascorbate deficiency is compensated by changes in the other major antioxidants. Seedlings grown in vitro were used to minimize stress and longer term developmental differences. Comparison was made with the low glutathione cad2 mutant and vtc2-1 treated with D,L-buthionine-[S,R]-sulphoximine to cause combined ascorbate and glutathione deficiency. The pool sizes and oxidation state of ascorbate and glutathione were not altered by deficiency of the other. alpha-Tocopherol and activities of monodehydroascorbate reductase, dehydroascorbate reductase, glutathione reductase, and catalase were little affected. Ascorbate peroxidase activity was higher in vtc1, vtc2-1, and vtc2-2. Ionically bound cell wall peroxidase activity was increased in vtc1, vtc2-1, and vtc4. Supplementation with ascorbate increased cell wall peroxidase activity. 2,6-Dichlorobenzonitrile, an inhibitor of cellulose synthesis, increased cell wall peroxidase activity in the wild type and vtc1. The transcript level of an endochitinase, PR1, and PR2, but not GST6, was increased in vtc1, vtc2-1, and vtc-2-2. Endochitinase transcript levels increased after ascorbate, paraquat, salicylic acid, and UV-C treatment, PR1 after salicylic acid treatment, and PR2 after paraquat and UV-C treatment. Camalexin was higher in vtc1 and the vtc2 alleles. Induction of PR genes, cell wall peroxidase activity, and camalexin in vtc1, vtc2-1, and vtc2-2 suggests that the mutants are affected in pathogen response signalling pathways.

Aluminum-Induced Cell Wall Peroxidase Activity and Lignin Synthesis Are Differentially Regulated by Jasmonate and Nitric Oxide

Cassia tora is an annual legume and cultivated as a traditional medicinal herb for multiple therapies including regulation of blood pressure and blood lipid. Because of naturally occurring acidic soils in southeastern China, this plant species may possess strategies for tolerance to low pH and aluminum toxicity. In the search for the regulatory basis of biochemical response to Al, cell wall-bound peroxidases, including lignin-generated peroxidases and NADH oxidases, were investigated in the root tips of C. tora. Activities of both types of peroxidases significantly increased with Al concentrations. Analysis with native PAGE also demonstrated the strong induction of cell wall peroxidases by Al. The Al-induced increasing activities of peroxidases were closely correlated with lignin accumulation and H 2O 2 production. The biochemical effect of exogenous nitric oxide (NO) and methyl jasmonic acid (MJ) was examined to investigate signal properties and lignin synthesis under Al stress. Application of MJ at 10 microM promoted root sensitivity to Al by activating apoplastic peroxidase activity and accumulating H 2O 2 and lignin, whereas the opposite action was found for NO. The sensitivity of apoplastic peroxidases under Al stress was associated with the cross-talk of MJ and NO signals. The analysis reveals that the activity of lipoxygenase (an enzyme for MJ biosynthesis), with its transcripts increased in Al-exposed roots, was depressed by NO exposure. The effect of MJ on intracellular NO production was also investigated. It is shown that NO staining with 4,5-diaminofluorescein diacetate fluorescence was intensified by Al but was suppressed by MJ. These results suggest that NO and MJ may interplay in signaling the cell wall peroxidase activity and lignin synthesis in the roots exposed to Al.

Cadmium induced oxidative stress and changes in soluble and ionically bound cell wall peroxidase activities in roots of seedling and 3–4 leaf stage plants of Brassica juncea (L.) czern

Metabolic adaptations to heavy metal toxicity in plants are thought to be related with developmental growth stage and the type of metal by which plant is affected. In the present study, changes in ionically bound CWP, soluble peroxidase activity, H(2)O(2) level and Malonaldehyde content in roots of cadmium and copper stressed seedlings and cadmium stressed 3-4 leaf stage plants of Brassica juncea were investigated. Cadmium inhibits root growth and reduces fresh biomass. The reduction in root growth and fresh biomass is correlated with increased lipid peroxidation and reduced tolerance. Treatment with cadmium resulted in an increase in ionically bound CWP activity in roots of seedlings but no significant change in its activity was found in roots of 3-4 leaf stage plants. Increased level of H(2)O(2) in roots of cadmium and copper treated seedlings, show a direct correlation with increased activity of ionically bound CWP. H(2)O(2) level in 3-4 leaf stage plant roots was found to be very low. Soluble peroxidase activity decreased in cadmium (50 and 100 mu-icroM) treated seedlings but it was ineffective to cause any change in its activity in 3-4 leaf stage plants. Copper treated seedlings showed an increase in ionically bound CWP activity, H(2)O(2) level and MDA content. Ascorbic acid (50 mM) pretreated seedlings shows significant decrease in ionically bound CWP activity when exposed to 50 muM cadmium. Hence, it is concluded that inhibition of root growth in Brassica juncea seedlings by cadmium, is associated with CWP catalyzed H(2)O(2) dependent reactions which are involved in metabolic adaptations to heavy-metal stress.

Comparative effects of regulated deficit irrigation (RDI) and partial root-zone drying (PRD) on growth and cell wall peroxidase activity in tomato fruits

The effects of regulated deficit irrigation (RDI) and partial root-zone drying (PRD) on tomato fruit growth and cell wall peroxidase activity in tomato exocarp were investigated in growth chamber conditions. The RDI treatment was 50% of water given to fully irrigated (FI) plants and the PRD treatment was 50% of water of FI plants applied to one half of the root system while the other half dried down, with irrigation shifted when soil water content of the dry side decreased 15–20%. RDI significantly reduced fruit diameter, though PRD reduced fresh weight while having no significant effect on fruit diameter. The activity of peroxidase was significantly higher in RDI and PRD treated plants compared to those of FI. Differences between RDI and PRD were expressed on temporal basis. In the fruits of RDI treated plants peroxidase activity began to increase in the phase when fruit growth started to decline with the peak of enzyme activity of 6.1 HRPEU g −1 FW reached in the phase of mature green fruits when fruit growth rate was minimal. Increase of peroxidase activity in PRD fruits coincided with the ripening phase and the peak of enzyme activity (5.3 HRPEU g −1 FW) was measured at the end of fruit ripening. These data potentially identified contrasting and different roles of tomato exocarp cell wall peroxidase in RDI and PRD treated plants. In RDI treated plants peroxidase may have a role in restricting fruit growth rate, although the increase in enzyme activity during ripening of PRD treated fruit pointed out that peroxidase may also control fruit maturation by inducing more rapid process.