The role of potassium phosphite in chlorophyll fluorescence and photosynthetic parameters of downy mildew-challenged cucumber Cucumis sativus plants

The effect of potassium phosphite (KPhi) on the photosynthetic parameters of cucumber plants inoculated with Pseudoperonospora cubensis was investigated in the present study. Cucumber plants were treated with KPhi before or after inoculation with P. cubensis and leaf samples were collected at different time courses for assessments. Results showed that in pathogen-inoculated plants Fv/Fm was decreased up to 3%. The rate of quantum photosynthetic was also decreased significantly in inoculated plants. Downy mildew led to a decrease in chlorophyll amount which in turn reduced the efficiency of photosystem II. In the KPhi-treated leaves, chlorophyll a and b decreased by 72% and 68%, respectively. Remarkable reduction in the efficiency of photosystem II as well as increased lipid membrane disruption, led to increased lipid peroxidation rate of the membranes up to 52%. The results of this study indicate the mitigating role of potassium phosphite in reducing the adverse effects of pathogen and maintaining the photosynthetic apparatus efficiency in cucumber plants.     

NO和钙信使系统在草酸诱导黄瓜叶片抗霜霉病中的作用

通过草酸及其与不同抑制剂亚甲基蓝、EGTA、氯丙嗪和Li+组合处理黄瓜叶片,研究了草酸与抑制剂不同处理组合方式对黄瓜叶片POD活性和叶片病情指数的影响,探讨NO、钙信使系统在草酸诱导叶片抗霜霉病中的作用.结果显示,10～70mmol/L草酸均能不同程度诱导黄瓜叶片POD活性的升高,提高叶片对黄瓜霜霉病的抗病性,降低叶片病情指数,并以30mmol/L效果最好.4种抑制剂分别与30mmol/L草酸同时或先于草酸处理,或草酸处理后一定时间再用抑制剂处理,均明显抑制黄瓜叶片POD活性的升高及病情指数的降低.研究表明,NO、Ca2+、钙调素(CaM)和磷酸肌醇均可能参与了草酸诱导黄瓜霜霉病抗性的信号转导过程.     

Role of salicylic acid and oxidative burst in expression of resistance to Alternaria blight

The accumulation of salicylic acid and H₂O₂ during pathogenic infection of mustard plants with Alternaria brassicae spores was investigated to understand the role of these two defense compounds in the expression of resistance. Comparisons were made between a susceptible Brassica juncea variety RH30 and a Brassica carinata variety HC1, which is known to be resistant. An oxidative burst was detected as in situ accumulation of H₂O₂, in both the Brassica spp. after pathogen application. However, H₂O₂ generation was extracellular in the resistant variety and both extra- and intracellular in the susceptible variety. Endogenous levels of SA increased over 2.5-fold in the resistant variety HC1 in response to pathogen application and this increase was observed only in conjugated SA levels. Pathogen application also led to an increase in the antioxidant enzymes, guaiacol-dependent peroxidase (GDP) and superoxide dismutase (SOD) in HC1. Exogenous SA application to leaves led to over threefold increase in the free and conjugated SA levels in both varieties. Pathogen application to the SA pretreated plants led to over 10-fold increase in endogenous SA levels in both varieties as compared to the levels in controls and this correlated with a decrease in disease symptoms in both species. SA appeared to regulate defense responses in Brassica spp. in a concentration-dependent manner. While 2.7-fold increase in endogenous SA levels (as seen in HC1) led to an induction of antioxidant enzymes, over 10-fold increases in endogenous SA levels (as seen after exogenous SA application in both varieties) brought about no induction of the antioxidant enzymes, probably because SA itself served as an antioxidant.     

Effect of glutamate on Pyricularia oryzae infection of rice monitored by changes in photosynthetic parameters and antioxidant metabolism

Considering the importance of blast caused by Pyricularia oryzae in the decrease of rice yield worldwide, this study aimed to assess the photosynthetic performance [leaf gas exchange and chlorophyll (Chl) a fluorescence parameters as well as the photosynthetic pigments concentration], the activities of antioxidant enzymes [ascorbate peroxidase, catalase (CAT), peroxidase (POX), superoxide dismutase (SOD), glutathione peroxidase (GPX), glutathione reductase (GR) and glutathione-S-transferase] and concentrations of hydrogen peroxide (H₂O₂) and malondialdehyde (MDA) in the leaves of rice plants non-supplied (−Glu) or supplied (+Glu) with glutamate (Glu) and non-infected or infected by P. oryzae. Blast severity was reduced in the leaves of +Glu plants. On the infected leaves of +Glu plants, the values for internal CO₂ concentration were lower while the values for net carbon assimilation rate, stomatal conductance as well as for the concentrations of Chl a, Chl b and carotenoids were higher in comparison to infected leaves of −Glu plants. The functionality of the photosynthetic apparatus was preserved in the infected leaves of +Glu plants. The activities of CAT, GPX, GR, POX and SOD increased in the infected leaves of both −Glu and +Glu plants compared to their non-inoculated counterparts, but their activities were lower for +Glu plants. The lower activity of these antioxidative enzymes was triggered by the reduced hydrogen peroxide concentration in the infected leaves of +Glu plants resulting in lower MDA concentration. It can be concluded that photosynthesis was less impaired in infected plants supplied with glutamate due to the lower biochemical constraints for CO₂ fixation. Moreover, there was a need for lower activity of reactive oxygen species scavenging enzymes in infected leaves of plants supplied with glutamate due to the lower oxidative stress imposed by P. oryzae infection.     

Priming of Nicotiana benthamiana antioxidant defences using brown seaweed extracts

Extracts from the brown seaweeds Cystoseira myriophylloides, Laminaria digitata and Fucus spiralis were evaluated as plant defence inducers against the wild fire disease of Nicotiana benthamiana. Seeds’ imbibition in aqueous seaweed extracts (SE) results in plants with reduced disease severity. In addition, bacterial populations were significantly reduced in these plants when compared to those germinated in sterile distilled water. They primed N. benthamiana for H₂O₂ accumulation and for enhanced activity of several antioxidant enzymes including catalase, ascorbate peroxidase, guaiacol peroxidase and polyphenol oxidase. These results revealed that soaking of seeds in SE before sowing allows N. benthamiana to reduce pathogen attack and can be easily applied in practice.     

Streptomyces spp. alleviate Rhizoctonia solani‐mediated oxidative stress in Solanum lycopersicon

The ability of Streptomyces species to act as biocontrol agents for plant pathogens via induced systemic resistance has been demonstrated and considerable efforts have been made in elucidating the underlying mechanisms of Streptomyces–host plant–phytopathogen interactions. Here, we have assessed the ability of Streptomyces coelicolor, Streptomyces griseus, Streptomyces albus, Streptomyces antibioticus and Streptomyces champavatii to provide disease protection against Rhizoctonia solani in Solanum lycopersicon and have also examined associated changes in hydrogen peroxide (H₂O₂) production, lipid peroxidation (LPO) and antioxidant enzymes. The production of H₂O₂ at the second day after pathogen inoculation (dapi) was observed to be 1.1‐fold higher in Streptomyces‐treated plants, when compared to untreated inoculated control plants. A similar increase in catalase and ascorbate peroxidase activity was observed at fourth dapi whereas increased activities of guaiacol reductase and glutathione peroxidase were observed at fifth dapi. Likewise, LPO reached a maximum at sixth dapi in untreated inoculated plants while in Streptomyces‐treated plants it was observed to be 1.3–1.5‐fold less when compared to untreated inoculated control plants. This study offers novel insights into the mechanisms of priming by Streptomyces and highlights their capacity to activate plant defence responses generated by biotic stress through induction of antioxidant enzymes along with improved reactive oxygen species management.     

Drought Induces Oxidative Stress and Enhances the Activities of Antioxidant Enzymes in Growing Rice Seedlings

When rice seedlings grown for 10 and 20 days were subjected to in vitro drought stress of −0.5 and −2.0 MPa for 24 h, an increase in the concentration of superoxide anion (O₂^.−), increased level of lipid peroxidation and a decrease in the concentration of total soluble protein and thiols was observed in stressed seedlings compared to controls. The concentration of H₂O₂ as well as ascorbic acid declined with imposition of drought stress, however glutathione (GSH) concentration declined only under severe drought stress. The activities of total superoxide dismutases (SODs) as well as ascorbate peroxidase (APX) showed consistent increases with increasing levels of drought stress, however catalase activity declined. Mild drought stressed plants had higher guaiacol peroxidase (GPX) and chloroplastic ascorbate peroxidase (c-APX) activity than control grown plants but the activity declined at the higher level of drought stress. The activities of enzymes involved in regeneration of ascorbate i.e. monodehydroascorbate reductase (MDHAR), dehydroascorbate reductase (DHAR) and glutathione reductase (GR) were higher in drought stressed plants compared to controls. Results suggest that drought stress induces oxidative stress in rice plants and that besides SOD, the enzymes of ascorbate-glutathione cycle, which have not been studied in detail earlier under stressful conditions, appear to function as important component of antioxidative defense system under drought stress.     

Effects of salicylic acid on paraquat tolerance in<Emphasis Type="Italic">Arabidopsis thaliana</Emphasis> plants

Foliar spraying ofArabidopsis thaliana (Columbia ecotype) plants with a 1.0-mM salicylic acid (SA) solution significantly improved their tolerance to subsequent paraquat (PQ)-induced oxidative damage. Leaf injuries, including losses of chlorophyll, protein, and fresh weight, were reduced. Our analysis of antioxidant enzymes in the leaves showed that SA pre-treatment effectively retarded rapid decreases in the activities of Superoxide dismutase (SOD), catalase, and ascorbate peroxidase that are normally associated with PQ exposure. In addition, guaiacol peroxidase activity was remarkably increased. In a native gel assay of peroxidase (POD) isozymes, staining activity of the POD1 isozyme, which disappeared in plants exposed only to 10 μM PQ, was significantly recovered by the 1.0-mM SA pre-treatment POD2 isozyme activity was also pronounced in all SA-treated plants compared with the control. A 12-h SA pre-treatment, without subsequent PQ stress, also caused a small increase in the endogenous H₂O₂ content that accompanies the symptoms of mild leaf injuries. This enhanced level occurred in parallel with a slight SOD increase and a catalase decrease. From our results, it can be assumed that, due to the small increase in SOD as well as catalase inactivation via SA pre-treatment, a moderate increase in H₂O₂ levels may occur. In turn, a large induction of guaiacol peroxidase leads to enhanced PQ tolerance inA. thaliana plants.     

The beneficial role of potassium in Cd-induced stress alleviation and growth improvement in Gladiolus grandiflora L.

Heavy metal contaminated agricultural soils are one of the most important constraints for successful cultivation of crops. The current research was conducted to evaluate the role of potassium (K) on plant growth and amelioration of cadmium (Cd) stress in Gladiolus grandiflora under greenhouse conditions. G. grandiflora corms were sown in media contaminated with 0 (C), 50 (Cd50) and 100 (Cd100) mg Cd kg^?1 soil. The plants growing in Cd-contaminated media exhibited reduced gas exchange attributes, chlorophyll (Chl) contents, vegetative and reproductive growth as compared to control. The plants raised in Cd contaminated media showed reduced nutrition yet higher Cd contents. However, supplementation of 60 mg Kg^?1 K in treated plants (C+K, Cd50+K and Cd100+K) improved quantity of total soluble protein and proline (Pro) along with activity of antioxidant enzymes including superoxide dismutase (SOD), catalase (CAT), peroxidase (POD) and ascorbate peroxidase (APX) under Cd stress. Similarly, K supplementation reduced the level of malondialdehyde (MDA) and hydrogen peroxide (H₂O₂) in treated plants. Potassium supplemented plants exhibited better vegetative and reproductive growth. The improved stress tolerance in K supplemented plants was attributed to the reduced quantity of MDA and H₂O₂, enhanced synthesis of protein, proline, phenols, flavonides and improved activity of antioxidant enzymes. The present research supports the application of K for alleviation of Cd stress in G. grandiflora.     

Efficacy of Foliar Application of Phosphates in Controlling Powdery Mildew Fungus on Field-Grown Winegrapes: Effects on Cluster Yield and Peroxidase Activity in Berries

Seven foliar applications of 0.025M K₂HPO₄ and KH₂PO₄ (both plus Tween 20) and the commercial systemic fungicides, Dorado (Pyrifenox) 480 EC, Penconazole EC and Benomyl, were applied at 14-day intervals starting at 10-cm shoot length on field-grown Chardonnay winegrapes. Both phosphates and systemic fungicides inhibited development of powdery mildew fungus (Uncinula necator, Schw., Burr.) on fruit clusters, as compared with untreated control vines. Diseases, everity on clusters of plants treated with K₂HPO₄ and fungicides was 0.3 and 0.2, respectively, as compared with 1.3 on control clusters (on a 0–4 scale), for the first rating, conducted 10 days after the fifth application of fungicides and phosphates, Five days after the last application, disease severity was 3.5 on non-treated control clusters and 0.3 and 0.8 on clusters treated with Dorado and K₂HPO₄, respectively. Powdery mildew infection remarkably reduced the weight of non-treated control clusters as compared with Dorado and phosphate treated clusters. Phosphate treatment caused an increase (3-fold) of peroxidase activity in the soluble fraction of non-infected control berries. A remarkable peroxidase enhancement was detected in the soluble (8-fold) and ionically bound (2-fold) fractions from the phosphate-treated and infected berries. Results indicate that phosphates can be used as foliar fertilizers for disease control in the field and that peroxidase might, be involved in the defense process.     

Hydrogen peroxide detoxifying enzymes show different activity patterns in host and non-host plant interactions with Magnaporthe oryzae Triticum pathotype

Wheat blast caused by the hemibiotroph fungal pathogen Magnaporthe oryzae Triticum (MoT) pathotype is a destructive disease of wheat in South America, Bangladesh and Zambia. This study aimed to determine and compare the activities of antioxidant enzymes in susceptible (wheat, maize, barley and swamp rice grass) and resistant (rice) plants when interacting with MoT. The activities of reactive oxygen species-detoxifying enzymes; catalase (CAT), ascorbate peroxidase (APX), glutathione peroxidase (GPX), glutathione S-transferase (GST), peroxidase (POX) were increased in all plants in response to MoT inoculation with a few exceptions. Interestingly, an early and very high activity of CAT was observed within 24 h after inoculation in wheat, barley, maize and swamp rice grass with lower H₂O₂ concentration. In contrast, an early and high accumulation of H₂O₂ was observed in rice at 48 hai with little CAT activity only at a later stage of MoT inoculation. The activities of APX, GST and POD were also high at an early stage of infection in rice. However, these enzymes activities were very high at a later stage in wheat, barley, maize and swamp rice grass. The activity of GPX gradually decreased with the increase of time in rice. Taken together, our results suggest that late and early inductions of most of the antioxidant enzyme activities occurs in susceptible and resistant plants, respectively. This study demonstrates some insights into physiological responses of host and non-host plants when interacting with the devastating wheat blast fungus MoT, which could be useful for developing blast resistant wheat.     

Induction of defence response against blister blight by calcium chloride in tea

Tea, the second most consumed beverage after water in the world, is produced from the processed tender leaves of tea plants (Camellia sinensis). Production of tea is hindered by various biotic and abiotic stresses. Among the biotic factors, blister blight disease caused by an obligate fungal pathogen, Exobasidium vexans Massee, is a serious problem to the tea industry. The present study was to evaluate the efficacy of elicitor calcium chloride (CaCl₂) in inducing resistance in tea plants against blister blight disease. During the pick time of blister incidence at Darjeeling tea garden, the application of CaCl₂ at a concentration of 1% resulted in disease inhibition around 71% over the control set. Treatment also resulted in the induction of defence enzymes like peroxidase, polyphenol oxidase, phenylalanine ammonia lyase, β-1,3-glucanase and higher phenol accumulation. Furthermore, the increase in defence molecules also correlated with increase in nitric oxide (NO) generation, a potent defence molecule in plant defence. The result suggests that CaCl₂ can used as a potential elicitor in the integrated disease management in organic tea cultivation.     

Involvement of G6PDH in heat stress tolerance in the calli from Przewalskia tangutica and Nicotiana tabacum

Glucose-6-phosphate dehydrogenase (G6PDH) has been implicated in supplying reduced nicotine amide cofactors for biochemical reactions and in modulating the redox state of cells. In this study, the role of G6PDH in thermotolerance of the calli from Przewalskia tangutica and tobacco (Nicotiana tabacum L.) was investigated. Results showed that Przewalskia tangutica callus was more sensitive to heat stress than tobacco callus. The activity of G6PDH and antioxidant enzymes (ascorbate peroxidase, catalase, peroxidase and superoxide dismutase) in calli from Przewalskia tangutica and tobacco increased after 40 °C treatment, although two calli exhibited a difference in the degree and timing of response to heat stress. When G6PDH was partially inhibited by glucosamine pretreatment, the antioxidant enzyme activities and thermotolerance in both calli significantly decreased. Simultaneously, the heat-induced H₂O₂ content and the plasma membrane NADPH oxidase activity were also reduced. Application of H₂O₂ increased the activity of G6PDH and antioxidant enzymes in both calli. Diphenylene iodonium, a NADPH oxidase inhibitor, counteracted heatinduced H₂O₂ accumulation and reduced the heat-induced activity of G6PDH and antioxidant enzymes. Moreover, exogenous H₂O₂ was effective in restoring the activity of G6PDH and antioxidant enzymes after glucosamine pretreatment. Western blot analysis showed that G6PDH gene expression in both calli was also stimulated by heat and H₂O₂, and blocked by DPI and glucosamine under heat stress. Taken together, under heat stress G6PDH promoted H₂O₂ accumulation via NADPH oxidase and the elevated H₂O₂ was involved in regulating the activity of antioxidant enzymes, which in turn facilitate to maintain the steady-state H₂O₂ level and protect plants from the oxidative damage.     

Effect of Arbuscular Mycorrhizal Inoculation on Salt-induced Nodule Senescence in <Emphasis Type="Italic">Cajanus cajan</Emphasis> (Pigeonpea)

Many physiological and biochemical plant processes affected by salt stress trigger premature nodule senescence and decrease their ability to fix nitrogen. The objective of this study was to evaluate the role of arbuscular mycorrhiza (AM) in moderating salt-induced premature nodule senescence in Cajanus cajan (L.) Millsp. Greenhouse experiments were conducted in which the plants were exposed to salinity stress of 4, 6, and 8 dSm⁻¹. Various parameters linked to nodule senescence were assessed at 80 days after sowing. Nodulation, leghemoglobin content, and nitrogenase enzyme activity measured as acetylene-reducing activity (ARA) were evaluated. Two groups of antioxidant enzymes were studied: (1) enzymes involved in the detoxification of O₂⁻ radicals and H₂O₂, namely, superoxide dismutase (SOD), catalase (CAT) and peroxidase (POX), and (2) enzymes that are important components of the ascorbate glutathione pathway responsible for the removal of H₂O₂, namely, glutathione reductase (GR) and ascorbate peroxidase (APOX). Exposure of plants to salinity stress enhanced nodule formation; however, nodule growth suffered remarkably and a marked decline in nodule biomass, relative permeability, and lipid peroxidation was observed. Leghemoglobin content and ARA were reduced under saline conditions. AM significantly improved nodulation, leghemoglobin content, and nitrogenase activity under salt stress. Activities of SOD, CAT, APOX, POX, and GR increased markedly in mycorrhizal-stressed plants. A synthesis of the evidence obtained in this study suggests a correlation between enhanced levels of antioxidant enzyme activities, reduced membrane permeability, reduced lipid peroxidation, and improved nitrogen-fixing efficiency of AM plants under stressed and unstressed conditions. These factors could be responsible for the protective effects of mycorrhiza against stress-induced premature nodule senescence.     

Application of Bacillus megaterium MCR-8 improved phytoextraction and stress alleviation of nickel in Vinca rosea

The current research was performed to evaluate the effect of Bacillus megaterium MCR-8 on mitigation of nickel (Ni) stress in Vinca rosea grown on Ni-contaminated soil (50, 100, and 200 mg Ni kg^?1 soil). The treated plants exhibited reduced growth, biomass, gas exchange capacity, and chlorophyll (Chl) content under Ni stress. The inoculated plants growing in Ni-contaminated media exhibited relatively higher growth, total soluble protein, and proline contents. Similarly, bacterial inoculation improved the activity of antioxidant enzymes including superoxide dismutase (SOD), catalase (CAT), peroxidase (POD), and ascorbate peroxidase (APX) under Ni stress. The Ni stress alleviation in inoculated plants was attributed to the reduced level of malondialdehyde (MDA) and hydrogen peroxide (H₂O₂), enhanced synthesis of protein, proline, phenols, and flavonides in conjunction with improved activity of antioxidant enzymes. The growth-promoting characteristics of microbe such as 1-aminocyclopropane-1-carboxylate deaminase (ACCD) and phosphate solubilization activity, siderophore, and auxin production capability also improved the growth and stress mitigation in inoculated plants. Furthermore, the inoculated plants exhibited higher value for bioconcentration factor (BCF), translocation factor (TF), and resulted in higher loss of Ni content from soil. The current results exhibited the beneficial role of B. megaterium MCR-8 regarding stress alleviation and Ni phytoextraction by V. rosea.     

Jasmonic Acid-Mediated-Induced Resistance in Groundnut (Arachis hypogaea L.) Against Helicoverpa armigera (Hubner) (Lepidoptera: Noctuidae)

Jasmonic acid (JA) acts as a signal molecule to induce resistance in plants against herbivores and its levels are elevated in plants after wounding or insect damage. Groundnut is an important crop in many tropical and subtropical regions worldwide, but there is surprisingly little knowledge on its induced defenses against herbivores. The effect of JA as a spray on induced resistance in three groundnut genotypes, namely, ICGV 86699 (resistant), NCAc 343 (resistant), and TMV 2 (susceptible), against Helicoverpa armigera was studied. The activity of oxidative enzymes [peroxidase (POD) and polyphenol oxidase (PPO)] and the amounts of other host plant defense components [total phenols, hydrogen peroxide (H₂O₂), malondialdehyde (MDA), and protein content] were recorded at 24, 48, 72, and 96 h after pretreatment (1 day) with JA followed by infestation with H. armigera (PJA + HIN) and H. armigera infestation with simultaneous JA application (HIN + JA) to understand the consequences of induced resistance in groundnut. The plant damage, larval survival, and larval weights were also recorded. There was a significant increase in POD and PPO activities and in the amounts of total phenols, H₂O₂, MDA, and proteins in PJA + HIN- and JA + HIN-treated plants as compared to the plants treated with JA and infested with H. armigera individually and to untreated control plants. Among all the genotypes, the strongest induction of defense was observed in the ICGV 86699 genotype. It is concluded that pretreatment with JA and its application during low levels of insect infestation can increase the levels of host plant resistance against herbivorous insects and reduce the pest-associated losses in groundnut.     

Study of antioxidant enzymes activity and isozymes pattern in hairy roots and regenerated plants in <Emphasis Type="Italic">Nicotiana tabacum</Emphasis>

Hairy root disease is caused by the infection of wounded higher plants with Agrobacterium rhizogenes. Transformation of tissues or plants with A. rhizogenes, and with rol genes, as well as hairy roots may produce alterations in the plant secondary metabolism. H₂O₂ and other ROS are involved as a signal in secondary metabolite production pathway and play a key role in plant defensive reactions. In this work, the effect of A. rhizogenes T-DNA on nicotine content, antioxidant enzymes activity, H₂O₂ production, pattern of peroxidase (POX) and superoxide dismutase (SOD) isozymes in hairy roots and regenerated plants were studied. Rise in SOD and POX activities in the transformed lines of TRa and TRb and in the resultant regenerated plants, also the decreased level of H₂O₂ in them, compared with the untransformed lines indicates that, the T-DNA genes expression of A. rhizogenes probably decreases H₂O₂ level by increasing the production of antioxidant enzymes. Decrees the level of H₂O₂ content in TRc line in spite of the similarity of antioxidant enzyme activity in comparison with normal root, indicate that A. rhizogenes activate other mechanisms except SOD and POX enzyme for reducing H₂O₂ level.     

Changes in the Activity of Antioxidant Enzymes in Wheat Leaves and Roots as a Function of Nitrogen Source and Supply

The activity of enzymes participating in the systems of antioxidant protection was assayed in the second leaf and roots of 21-day-old wheat seedlings (Triticum aestivum L.) grown in a medium with nitrate (NO^– ₃ treatment), ammonium (NH⁺ ₄ treatment), or without nitrogen added (N-deficiency treatment). The activities of superoxide dismutase (SOD), peroxidase, ascorbate peroxidase, glutathione reductase, and catalase in the leaves and roots of the NH⁺ ₄ plants was significantly higher than in the plants grown in the nitrate medium. The activity of SOD decreased and ascorbate peroxidase markedly increased in leaves, whereas the activity of ascorbate peroxidase increased in the roots of N-deficient plants, as compared to the plants grown in nitrate and ammonium. Low-temperature incubation (5°, 12 h) differentially affected the antioxidant activity of the studied plants. Whereas leaf enzyme activities did not change in the NH⁺ ₄ plants, the activities of SOD, peroxidase, ascorbate peroxidase, and catalase markedly increased in the NO^– ₃ plants. In leaves of the N-deficient plant, the activity of SOD decreased; however, the activity of other enzymes increased. In response to temperature decrease, catalase activity increased in the roots of NO^– ₃ and NH⁺ ₄-plants, whereas in the N-deficient plants, the activity of peroxidase increased. Thus, in wheat, both nitrogen form and nitrogen deficiency changed the time-course of antioxidant enzyme activities in response to low temperature.     

Purification and stability of peroxidase of African oil palm Elaies guineensis

In the previous work, after screening tropical plants (43 species) for peroxidase activity, high activity has been detected in leaves of some palms and especially African oil palm Elaeis guineensis. This palm is widely cultivated in Colombia and presents a promising source for the industrial production of peroxidase. The initial enzyme isolation included homogenization and extraction of pigments using aqueous two phase polymer system. Initially, traditional system, formed by polyethyleneglycol/K₂HPO₄, was used. The replacement of K₂HPO₄ with (NH₄)₂SO₄ allowed direct application of the salt phase with accumulated peroxidase on a Phenyl-Sepharose column. The final purification was carried out by liquid chromatography on Sephacryl S200 and DEAE-Toyopearl columns. The specific activity of the purified peroxidase measured toward guaiacol was 4300 units per mg of protein. The molecular weight and isoelectric point for palm peroxidase were 57.000 and 3.8, respectively. Palm peroxidase possesses uniquely high thermostability and is more stable in organic solvents than horseradish peroxidase is.     

Production of reactive oxygen species and antioxidant enzymes of pea and soybean plants under hypoxia and high CO<Subscript>2</Subscript> concentration in medium

Generation of reactive oxygen species (ROS) and activities of antioxidant enzymes (catalase, peroxidase, ascorbate peroxidase) in pea (Pisum sativum L.) and soybean (Glycine max L.) under hypoxia (3–24 h) and high CO₂ concentration in medium were studied. In sensitive to hypoxia pea seedlings, hypoxia enhanced markedly production of superoxide anion-radical, hydroperoxides, and especially hydrogen peroxide. In more tolerant soybean plants, these changes were less pronounced. During first hours of hypoxia, activity of lipoxygenase in plant cells increased. This allows a suggestion that this enzyme is involved in the processes of hydroperoxide accumulation in plant tissues under oxygen deficit. In pea and soybean plants, a correlation between tolerance to hypoxia, the rate of ROS generation, and antioxidant enzyme activities was established. During the first hours of hypoxia, the catalase activity in soybean plants increased stronger than in sensitive to hypoxia pea plants. At longer exposure to hypoxia (24 h), peroxidases started to play the higher role in cell defense against hypoxia, but only in soybean plants. The medium with the higher CO₂ content induced higher changes in the processes of ROS accumulation and activities of lipoxygenase and antioxidant enzymes. This permits us to refer CO₂, accumulated as a product of respiration in the cells, to low-molecular signal molecules switching on plant adaptation to hypoxic stress.