Apoplastic Peroxidase Activity Research Articles

Putative NAD(P)-Binding Rossmann Fold Protein Is Involved in Chitosan-Induced Peroxidase Activity and Lipoxygenase Expression in Physcomitrium patens.

Oxidative burst, the rapid production of high levels of reactive oxygen species in response to external stimuli, is an early defense reaction against pathogens. The fungal elicitor chitosan causes an oxidative burst in the moss Physcomitrium patens (formerly Physcomitrella patens), mainly due to the peroxidase enzyme Prx34. To better understand the chitosan responses in P. patens, we conducted a screen of part of a P. patens mutant collection to isolate plants with less peroxidase activity than wild-type (WT) plants after chitosan treatment. We isolated a P.patens mutant that affected the gene encoding NAD(P)-binding Rossmann fold protein (hereafter, Rossmann fold protein). Three Rossmann fold protein-knockout (KO) plants (named Rossmann fold KO lines) were generated and used to assess extracellular peroxidase activity and expression of defense-responsive genes, including alternative oxidase, lipoxygenase (LOX), NADPH oxidase, and peroxidase (Prx34) in response to chitosan treatment. Extracellular (apoplastic) peroxidase activity was significantly lower in Rossmann fold KO lines than in WT plants after chitosan treatments. Expression of the LOX gene in Rossmann fold KO plants was significantly lower before and after chitosan treatment when compared with WT. Peroxidase activity assays together with gene expression analyses suggest that the Rossmann fold protein might be an important component of the signaling pathway leading to oxidative burst and basal expression of the LOX gene in P. patens. [Formula: see text] Copyright © 2023 The Author(s). This is an open access article distributed under the CC BY-NC-ND 4.0 International license.

Characterization of candidate genes for ozone tolerance in winter wheat (Triticum aestivum L.) and associated physiological mechanisms

Rising tropospheric ozone causes global yield reduction in wheat and therefore necessitates the breeding of ozone-resistant wheat varieties. The aim of this study was to explore contrasting physiological responses and characterize candidate genes for ozone tolerance in wheat identified in a genome-wide association study. Four wheat genotypes were grouped into tolerant and sensitive haplotypes according to their genetic backgrounds at ozone tolerant loci, and were investigated under season-long ozone fumigation in open-top chambers. Only sensitive haplotypes showed a significant decrease in net photosynthetic rate and a significant increase in lipid peroxidation indicating oxidative stress. Moreover, tolerant and sensitive haplotypes of wheat showed consistently contrasting antioxidant responses to ozone in terms of apoplastic ascorbate, ascorbate peroxidase and peroxidase activity. Candidate genes located within an ozone tolerant locus on chromosome 5A 592.04–593.33 Mb were analyzed through gene expression. The gene TraesCS5A01G400500 putatively involved in peroxidase activity was only upregulated in tolerant haplotypes. The differential regulation was consistent with sequence polymorphisms in the promoter region, where the cis-elements, EVENINGAT and PREATPRODH were only present in tolerant haplotypes. Based on the consistency of differential expression and sequence polymorphisms between tolerant and sensitive haplotypes, TraesCS5A01G400500 might explain contrasting physiological responses after ozone fumigation. This study demonstrated consistent transcriptional regulation, sequence polymorphisms in a candidate gene and physiological responses to ozone in different haplotypes, which represents a step forward in the understanding of the molecular mechanism underlying ozone tolerance in wheat.

Respiratory burst oxidases and apoplastic peroxidases facilitate ammonium syndrome development in Arabidopsis

Ammonium-nitrogen (NH4+) nutrition is linked to metabolic over-reduction for plants. The characteristic symptom of sole NH4+ nutrition is growth suppression, signifying this condition as the ammonium syndrome. In the present study, we investigated the mechanism of perception of high NH4+ conditions in Arabidopsis thaliana plants by examining apoplastic reactive oxygen species (ROS) metabolism. Major enzyme activity and a special pattern of expression of NADPH-dependent respiratory burst oxidases (RBOH) was found in Arabidopsis individuals cultured under NH4+ as the sole nitrogen source. This oxidative burst is independent of RBOHD/F expression and does not activate typical intracellular signalling pathways. In addition, elevated superoxide dismutase and apoplastic secretory peroxidase activities contributed to hydrogen peroxide (H2O2) accumulation in plants exposed to NH4+ nutrition. Consequently, higher H2O2 contents were determined in the extracellular space and were localised cytochemically. H2O2 is a substrate for cell wall cross-linking peroxidases, which showed enhanced activity in the presence of NH4+. Increase of cell wall polymerisation, could in turn inhibit cell elongation and slow down growth, as observed under NH4+ toxicity.

Photoperiod stress induces an oxidative burst-like response and is associated with increased apoplastic peroxidase and decreased catalase activities

Periodic changes of light and dark regulate numerous processes in plants. Recently, a novel type of stress caused by an extended light period has been described in Arabidopsis thaliana and was named photoperiod stress. Although photoperiod stress causes the induction of numerous stress response genes of which many are indicators of oxidative stress, the exact timing and mechanisms involved in dealing with this stress have not yet been investigated. We describe the response of the cellular redox system in wild-type Arabidopsis, the photoperiod stress sensitive cytokinin receptor mutant ahk2 ahk3 and the clock mutant cca1 lhy. Photoperiod stress caused several changes in the ROS scavenging system including a reduction of the ascorbic acid (AsA) redox status and strong peroxide formation during the night following the extended photoperiod. The changes were associated with reduced catalase (CAT) and increased apoplastic peroxidase (PRX) activities. Consistently, the expression of the apoplastic PRX genes PRX4, PRX33, PRX34 and PRX71 was strongly induced by photoperiod stress. We show that extending the light period by only few hours causes a stress response during the following night suggesting that the photoperiod stress response might occur in a natural setting.

Changes in apoplastic peroxidase activity and cell wall composition are associated with cold-induced morpho-anatomical plasticity of wheat leaves.

Temperate grasses, such as wheat, become compact plants with small thick leaves after exposure to low temperature. These responses are associated with cold hardiness, but their underlying mechanisms remain largely unknown. Here we analyse the effects of low temperature on leaf morpho-anatomical structure, cell wall composition and activity of extracellular peroxidases, which play key roles in cell elongation and cell wall thickening, in two wheat cultivars with contrasting cold-hardening ability. A combined microscopy and biochemical approach was applied to study actively growing leaves of winter (ProINTA-Pincén) and spring (Buck-Patacón) wheat developed under constant warm (25°C) or cool (5°C) temperature. Cold-grown plants had shorter leaves but longer inter-stomatal epidermal cells than warm-grown plants. They had thicker walls in metaxylem vessels and mestome sheath cells, paralleled with accumulation of wall components, predominantly hemicellulose. These effects were more pronounced in the winter cultivar (Pincén). Cold also induced a sharp decrease in apoplastic peroxidase activity within the leaf elongating zone of Pincén, and a three-fold increase in the distal mature zone of the leaf. This was consistent with the enhanced cell length and thicker cell walls in this cultivar at 5°C. The different response to low temperature of apoplastic peroxidase activity and hemicellulose between leaf zones and cultivar types suggests they might play a central role in the development of cold-induced compact morphology and cold hardening. New insights are presented on the potential temperature-driven role of peroxidases and hemicellulose in cell wall dynamics of grasses.

A saponin isolated from Agapanthus africanus differentially induces apoplastic peroxidase activity in wheat and displays fungicidal properties

A spirostane with an attached trisaccharide, (25R)-5α-spirostane-2α,3β,5α-triol 3-O-(O-α-l-rhamnopyranosyl-(1 → 2)-O-(β-d-galactopyranosyl-(1 → 3))-β-d-glucopyranoside), was isolated and identified from the aerial parts of Agapanthus africanus by activity-guided fractionation. Fungicidal properties of the crude extract, semi-purified fractions as well as the purified active saponin from A. africanus were screened in vitro against Fusarium oxysporum. At a concentration of 1 mg mL−1, the crude extract and semi-purified ethyl acetate and dichloromethane fractions showed significant antifungal activity. The purified saponin inhibited the in vitro mycelial growth of F. oxysporum completely (100 %) at a concentration of 125 µg mL−1. Furthermore, to verify previously observed induced resistance by crude extracts of A. africanus towards leaf rust, intercellular PR-protein activity was determined in wheat seedlings following foliar application of the purified saponin at 100 µg mL−1. In vitro peroxidase enzyme activity increased significantly (60 %) in wheat seedlings 48 h after treatment with the purified saponin, demonstrating its role as an elicitor to activate a defence reaction in wheat.

Endogenous ascorbate restrains apoplastic peroxidase activity during sunflower leaf development

Several apoplastic enzymes have been implicated in the control of elongation growth of plant cells. Among them, peroxidases contribute to both loosening and stiffening of the cell wall. They appear to be regulated by various mechanisms, including the action of extracellular inhibitors. To obtain evidence of the role of the enzyme-inhibitor interaction during leaf development, the intercellular washing fluids from Helianthus annuus leaves of different ages were isolated using standard methods of vacuum infiltration and centrifugation. Peroxidase activities, assessed using tetramethylbenzidine as substrate, increased during leaf development, reaching a maximum value after the leaves were fully expanded. An inhibitor, chemically characterised as ascorbate, co-localised with the enzyme in the apoplast. Moreover, there was a strong negative correlation between the action of peroxidase and the micromolar concentration of ascorbate in the apoplastic fluid. The results show that in growing leaves, the in planta ascorbate concentration is able to restrain peroxidase enzyme activity. Then, at the time of growth cessation, the loss of extracellular ascorbate relieves the inhibition on this enzyme that contributes to wall fixation.

Oxidative reactions in axenically seedling roots of olive (Olea europaea, L.)

The production of reactive oxygen species (ROS) plays important roles in the life cycle and in the stress response and defence mechanisms of plants. Various enzyme systems are involved in the formation of ROS in the apoplast, including plasmalemma NADPH oxidase and apoplastic peroxidases. The production of O 2 ·− and apoplastic peroxidase and exogenous NADH oxidation activities are all strongly dependent on the age of roots—the younger the root, the greater the activity. Apoplastic production of ROS is shown in the root by using specific histochemical probes, this ROS production is growing zone dependent. In the present study, using olive seedlings, differences were also observed between cultivars, especially in O 2 ·− production by the Verdial cultivar which was well above that of other cultivars studied. In all the cultivars, treatment of roots with methyl jasmonate (MeJA) or methyl salicylate (MeSA) increased O 2 ·− production. Similar results were observed for peroxidase activity, but not for the oxidation of exogenous NADH which was either unaffected (MeJA) or even partially inhibited (MeSA). A conclusion was that MeJA or MeSA induced apoplastic production of ROS does not use exogenous NADH. Treatment with diphenylene iodonium (DPI) reduced the formation of O 2 ·− , but affected neither peroxidase nor NADH oxidation activities. Cyanide inhibited O 2 ·− production and peroxidase and NADH oxidation activities. Treatment with MnCl2 had a strong stimulatory effect on peroxidase and NADH oxidation activities, but much less on O 2 ·− production. Finally, azide greatly reduced all activities, but especially O 2 ·− production. Together, these results indicate a relationship between oxidative activities and the processes of root growth, and that those activities are also dependent on the cultivar, as well as an involvement of peroxidases and plasmalemma NADPH oxidase in apoplast ROS production which is sensitive to DPI, azide, and cyanide but relatively insensitive to MnCl2, while exogenous NADH oxidation is linked to peroxidase activity.

Apoplastic superoxide production and peroxidase activity by intact and excised axenically grown seedling roots of sunflower

Excised and cold-preincubated sunflower seedling roots were compared with intact non-preincubated roots to test the effect of the injury stress and cold preincubation on the oxidative burst measured as apoplastic superoxide (O (2) (.-) ) generation and exocellular peroxidase (ECPOX) activity. Preincubated excised or intact roots released into the medium apoplastic proteins with peroxidase activity. Intact and excised roots responded to methyl jasmonate by an immediate oxidative burst that could not be induced by salicylic acid; both phytohormones also induced a slight and slow O (2)(.-) generation and ECPOX activity on excised roots, when added to the cold preincubation medium. The results with cyanide, azide, SHAM (ECPOX inhibitors) and diphenylene iodonium (inhibitor of trans-plasma membrane NAD(P)H-oxidases (NOX)-respiratory burst oxidase homologue in plants (RBOH), the trans-plasmamembrane nicotinamide adenine dinucleotide phosphate oxidase) are consistent with the hypothesis that different systems may be the origin of O (2) (.-) in intact and excised roots; ECPOX was an important component of them in both, together with NOX-RBOH in intact roots, but in excised roots the last one was replaced by an oxidase sensitive to the same inhibitors as the alternative mitochondrial oxidase. According to our hypothesis, these results could be explained if the electron flux would be deviated to different interconnected plasma membrane-redox systems, with different terminal oxidases, activated by different effectors or stresses.

Involvement of calcium-mediated effects on ROS metabolism in the regulation of growth improvement under salinity

Salinity reduces Ca 2+ availability, transport, and mobility to growing regions of the plant and supplemental Ca 2+ is known to reduce salinity damages. This study was undertaken to unravel some of the ameliorative mechanisms of Ca 2+ on salt stress at the cellular and tissue levels. Zea mays L. plants were grown in nutrient solution containing 1 or 80 mM NaCl with various Ca 2+ levels. Measurements of growth and physiological parameters, such as ion imbalance, indicated that the Ca 2+-induced alleviation mechanisms differed between plant organs. Under salinity, H 2O 2 levels increased in the leaf-growing tissue with increasing levels of supplemental Ca 2+ and reached the levels of control plants, whereas superoxide levels remained low at all Ca 2+ levels, indicating that Ca 2+ affected growth by increasing H 2O 2 but not superoxide levels. Salinity completely abolished apoplastic peroxidase activity. Supplemental Ca 2+ increased its activity only slightly. However, under salinity, polyamine oxidase (PAO) activity was shifted toward the leaf base probably as an adaptive mechanism aimed at restoring normal levels of reactive oxygen species (ROS) at the expansion zone where NADPH oxidase could no longer provide the required ROS for growth. Interestingly, addition of Ca 2+ shifted the PAO-activity peak back to its original location in addition to its enhancement. The increase in PAO activity in conjunction with low levels of apoplastic peroxidase is supportive of cellular growth via nonenzymatic wall loosening derived by the increase in H 2O 2 and less supportive of the peroxidase-mediated cross-linking of wall material. Thus extracellular Ca 2+ can modulate ROS levels at specific tissue localization and developmental stages thereby affecting cellular extension.

Extraction of Root Apoplastic Wall Fluid for Apoplastic Peroxidase Activity Assay

Plant roots secret a lot of peroxidases to counteract environmental influences. This protocol describes a way to extract root apoplastic wall fluid from Arabidopsis plants and to determine the peroxidase activity using guaiacol as substrate., 植物根秘密很多过氧化物酶抵抗环境影响。 这个协议描述了一种从拟南芥植物提取根体外形成壁液体和使用愈创木酚作为底物确定过氧化物酶活性的方法。

Wound‐induced apoplastic peroxidase activities: their roles in the production and detoxification of reactive oxygen species

Production of reactive oxygen species (ROS) is a widely reported response of plants to wounding. However, the nature of enzymes responsible for ROS production and metabolism in the apoplast is still an open question. We identified and characterized the proteins responsible for the wound-induced production and detoxification of ROS in the apoplast of wheat roots (Triticum aestivum L.). Compared to intact roots, excised roots and leachates derived from them produced twice the amount of superoxide (O2(*-)). Wounding also induced extracellular peroxidase (ECPOX) activity mainly caused by the release of soluble peroxidases with molecular masses of 37, 40 and 136 kD. Peptide mass analysis by electrospray ionization-quadrupole time-of-flight-tandem mass spectrometry (ESI-QTOF-MS/MS) following lectin affinity chromatography of leachates showed the presence of peroxidases in unbound (37 kD) and bound (40 kD) fractions. High sensitivity of O2(*-)-producing activity to peroxidase inhibitors and production of O2(*-) by purified peroxidases in vitro provided evidence for the involvement of ECPOXs in O2(*-) production in the apoplast. Our results present new insights into the rapid response of roots to wounding. An important component of this response is mediated by peroxidases that are released from the cell surface into the apoplast where they can display both oxidative and peroxidative activities.

Cadmium-induced accumulation of hydrogen peroxide in the leaf apoplast of Phaseolus aureus and Vicia sativa and the roles of different antioxidant enzymes

The effects of cadmium (Cd) on the accumulation of hydrogen peroxide (H 2O 2) and superoxide anion (O 2 −) in leaves of Phaseolus aureus and Vicia sativa were investigated. Cadmium at 100 μM significantly increased the production of O 2 − and H 2O 2, as well as the activities of plasma membrane-bound nicotinamide adenine dinucleotide phosphate (NADPH) oxidases and the symplastic and apoplastic activities of superoxide dismutase and ascorbate peroxidase in the leaves of both species. Apoplastic guaiacol peroxidase activity was significantly induced in the leaves of both species, particularly in P. aureus exposed to 100 μM Cd. Experiments with diphenylene iodonium as an inhibitor of NADPH oxidase and NaN 3 as an inhibitor of peroxidase showed that the majority of Cd-induced reactive oxygen species production in the leaves of both species may involve plasma membrane-bound NADPH oxidase and apoplastic peroxidase. Compared to V. sativa, increases in Cd-induced production of O 2 − and H 2O 2 and activities of NADPH oxidase and apoplastic peroxidase were more pronounced in P. aureus. In contrast, V. sativa had higher leaf symplastic superoxide dismutase and ascorbate peroxidase activities than P. aureus. The results indicated that V. sativa was more tolerant to Cd than P. aureus.

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.

Apoplastic antioxidative system responses to ozone stress in strawberry leaves

Cell wall polysaccharides, pectin composition, as well as apoplastic superoxide dismutase and peroxidase activities were investigated in strawberry (Fragaria x ananassa) cultivars (cvs) Korona and Elsanta differing in their ozone sensitivity. Plants were exposed to 140-170 microg m(-3) ozone either short-term for 7 days or long-term for 2 months in order to investigate whether differences in ozone sensitivity were due to differences in the apoplastic antioxidative systems. Cell wall polysaccharides were increased after 7 days and 2 months of ozone stress. While water-soluble pectins, low methoxy pectinates and NaOH-soluble pectinates were elevated after 7 days, their contents were unaffected (water-soluble pectins) or lower (low methoxy pectinates, NaOH-soluble pectinates) after 2 months. In cv. Elsanta, ozone treatment resulted in a significant reduction of superoxide dismutase activity after 7 days and 2 months, while it remained similar in cv. Korona. After 7 days, peroxidase activity was significantly higher in ozone-exposed leaves of cv. Korona, whereas after 2 months it was similar to or higher than in controls. Superoxide dismutase in cv. Korona detoxified ozone and its products in the apoplast, and the resulting elevated levels of H(2)O(2) were balanced within 7 days by an increase in peroxidase activity. Long-term peroxidase activity may not play a comparably significant role in ozone defence, but the increase in cell wall polysaccharides and cell wall thickness measured after 2 months, resulting in a decrease in specific leaf area, reflected structural modifications that limited activities of reactive oxygen species efficiently. In contrast, the reduction of superoxide dismutase activity in cv. Elsanta indicated a less efficient apoplastic radical scavenging system, at least during the first 7 days of ozone stress, which was accompanied by membrane leakage and contributed to accelerated leaf senescence. Long-term, the reduction of intercellular air space volume in leaves contributed to ozone tolerance of cv. Elsanta as in cv. Korona.

Apoplastic anti-oxidants in pepper (Capsicum annuum L.) fruit and their relationship to blossom-end rot

SummaryThe aim of the present research was to study changes in the major anti-oxidative enzyme activities known to be associated with the apoplast, during the induction of blossom-end rot (BER) in bell pepper (Capsicum annuum L.) fruits grown under saline conditions. Pepper plants of the BER-sensitive cv. ‘Mazurka’ and less-sensitive cv. ‘Selica’ were grown under control, low- and high-salinity irrigation regimes. Fruits were harvested continuously and, after approx. 7 months under these conditions, BER symptoms started to appear (in the Spring season) and fruits were sampled for biochemical measurements. The intercellular washing fluid (IWF; apoplast fraction) of the fruit pericarp was prepared and enzyme activities in that fraction were determined. The results showed that production of reactive oxygen species (ROS) in the fruit apoplast was higher in ‘Mazurka’ than in ‘Selica’. Apoplastic peroxidase and superoxide dismutase (SOD) activities, and the concentration of ascorbic acid, increased during ‘Selica’ fruit development, whereas the concentration of H2O2 decreased. During the stage of rapid growth, when fruits are most susceptible to BER, apoplastic peroxidase activity was greatly enhanced in the healthy pericarp of BER-affected fruits compared to healthy fruits. Under saline conditions, apoplastic peroxidase and SOD activities, as well as ascorbic acid and H2O2 levels, increased in healthy fruits of the resistant cultivar ‘Selica’; whereas, in healthy fruit of the BER-sensitive cultivar ‘Mazurka’, apoplastic peroxidase activity decreased while the level of ascorbic acid increased, but to a much lesser extent than in ‘Selica’. SOD activity and H2O2 levels increased similarly in both cultivars with increasing levels of salinity. It is proposed that apoplast-associated peroxidase activity and ascorbic acid concentration are increased coordinately in response to salt stress and may be part of a protective anti-oxidation mechanism that determines cultivar sensitivity to BER.

Changes in intracellular and apoplastic peroxidase activity, ascorbate redox status, and root elongation induced by enhanced ascorbate content in Allium cepa L.

Onions (Allium cepa L.) treated with external ascorbic acid or with the immediate precursor of its synthesis L-galactono-gamma-lactone show a stimulated elongation rate of the roots and an increase in the number of new radicles appearing at the bulb base. Treatment with both molecules resulted in an enhanced accumulation of ascorbate and dehydroascorbate along the root axis, but the distribution of these redox forms was not uniform along the root, as detected in intracellular (symplastic) and extracellular (apoplastic) compartments. Thus, those radicular zones metabolically more active, such as the meristem and the elongation zone, accumulated the highest amount of both redox forms of ascorbate. On the other hand, ascorbate and L-galactono-gamma-lactone also stimulated cytosolic glucose-6-phosphate dehydrogenase activity and inhibited peroxidase activity as deduced from in vivo and in vitro experiments. Differences were also found when comparing apoplastic and symplastic activities. These results are compatible with the idea of an ascorbate-mediated stimulation of root growth by inhibiting cell wall stiffening and increasing root metabolism.

Relationship of growth cessation with the formation of diferulate cross-links and p-coumaroylated lignins in tall fescue leaf blades.

We examined relationships among cell wall feruloylation, diferulate cross-linking, p-coumarate deposition, and apoplastic peroxidase (EC 1.11.1.7) activity with changes in the elongation rate of leaf blades of slow and rapid elongating genotypes of tall fescue ( Festuca arundinacea Schreb.). Growth was not directly influenced by ferulic acid deposition but leaf elongation decelerated as 8-5-, 8- O-4-, 8-8-, and 5-5-coupled diferulic acids accumulated in cell walls. Growth rapidly slowed and stopped with the deposition of p-coumarate, which is primarily associated with lignification in grass cell walls. Accretion of ferulate, diferulates and p-coumarate continued after growth ended, into the later stages of secondary wall formation. The concentration of 8-coupled diferulates dwarfed that of the more commonly measured 5-5-coupled isomer, suggesting that the latter dimer is a poor indicator of diferulate cross-linking in cell walls. Further work is required to clearly demonstrate the role of diferulate cross-linking and p-coumaroylated lignins in the cessation of leaf growth in grasses.

Growth Performance and Lignification in Tobacco with Suppressed Apoplastic Anionic Peroxidase Activity under Ambient and Elevated CO2 Concentrations

To study the relationship between growth, lignification, and peroxidase (POD) activity, we grew tobacco (Nicotiana tabacum var. Xanthi) control and transgenic plants, repressed in the anionic apoplastic POD by antisense transformation, under ambient and elevated CO2. It was expected that, if POD in antisense tobacco was at the threshold for normal lignification, POD activity would limit lignin production when growth was stimulated, thereby leading to diminished lignin concentrations under elevated CO2. If the maintenance of normal POD activities was associated with metabolic costs that can be reallocated to growth in plants with suppressed POD activities, elevated CO2 was expected to stimulate growth significantly more in antisense‐POD tobacco than in controls. Transgenic tobacco displayed 15% taller stems at bud set than controls, but this had no effect on whole‐plant biomass. There was no extra effect of elevated CO2 on biomass production in antisense‐POD plants as compared with untransformed plants. Th...

Gibberellic acid and dwarfism effects on the growth dynamics of B73 maize ( Zea mays L.) leaf blades: a transient increase in apoplastic peroxidase activity precedes cessation of cell elongation

The relationship between apoplastic peroxidase (EC 1.11.1.7) activity and cessation of growth in maize ( Zea mays L.) leaf blades was investigated by altering elongation zone length. Apoplastic peroxidase activity in the elongation and secondary cell wall deposition zones of elongating leaf blades of the maize inbred line B73 was used as a control and compared to leaves of the dwarf mutant D8‐81127, a near‐isogenic line of B73 unresponsive to gibberellins, and to leaves of B73 plants to which gibberellic acid (GA 3 ) had been applied via root uptake. Elongation zone length was increased by treatment with GA 3 through an increase in cell number as well as increased final cell length. The shorter elongation zone of dwarf leaves occurred primarily through reduced final cell length. Although elongation zone length differed among dwarf, control, and GA 3 ‐treated leaf blades, in all three treatments a transient increase in apoplastic peroxidase activity preceded a reduction in the segmental elongation rate in leaves. A peroxidase isoenzyme with pI 7.0 occurred in the leaf elongation zone during growth deceleration in all three treatments, and its activity decreased as growth displaced tissue into the region of secondary cell wall deposition. Growth cessation for all treatments coincided with the first appearance of peroxidase isozymes with pIs of 5.6 and 5.7. Based on the activity of particular isozymes relative to growth and differentiation, the pI 7.0 isoenzyme is most likely to be involved in cessation of cell elongation, while isozymes with pIs 5.6 and 5.7 are likely to be active in lignification.