In vivo and in vitro protective role of proline

Accumulation of proline in response to environmental stresses seems tobe widespread among plants. To elucidate the role of proline in plantresponses,in vivo and in vitro, we studied theeffect of proline on catalase (CAT; EC 1.11.1.6), peroxidase (POD; EC 1.11.1.7)and polyphenol oxidase (PPO; EC 1.14.18.1). In vivo, thesethree enzymes were activated by proline, while CAT and POD were activated andPPO was inactivated by NaCl. In vitro, CAT and POD wereactivated and PPO was inactivated by proline. Proline appeared to protect thesethree enzyme activities. The significance of these findings with regard toenvironmental stress-induced proline accumulation in vivois discussed. The ability of proline to activate the enzymes may suggest alimited conformational change. These results are important for characterisationof metabolic responses to environmental stresses and can be used as a stressindicator.     

Exogenous proline mitigates the detrimental effects of salt stress more than exogenous betaine by increasing antioxidant enzyme activities

Proline and betaine accumulate in plant cells under environmental stresses including salt stress. Here, we investigated effects of proline and betaine on the growth and activities of antioxidant enzymes in tobacco Bright Yellow-2 (BY-2) culture cells in suspension under salt stress. Both proline and betaine mitigated the inhibition of growth of BY-2 cells under salt stress and the mitigating effect of proline was more than that of betaine. Salt stress significantly decreased the activities of superoxide dismutase (SOD), catalase and peroxidase in BY-2 cells. Exogenous application of proline or betaine alleviated the reduction in catalase and peroxidase activities but not SOD activity under salt stress. In addition, proline was found to be effective in alleviating the inhibition of salt stress-induced catalase and peroxidase activities in BY-2 cells. Neither proline nor betaine directly scavenged superoxide (O(2)(-)) or hydrogen peroxide (H(2)O(2)). It is concluded that exogenous proline mitigates the detrimental effects of salt stress more than exogenous betaine because of its superior ability to increase the activities of antioxidant enzymes.     

玉米幼苗抗氧化酶活性、脯氨酸含量变化及与其耐寒性的关系

研究了黄C（耐寒型）和Mo17（低温敏感型）两种耐寒性不同的玉米自交系幼苗在低温逆境（5 ℃,3 d）下,根、中胚轴和胚芽鞘3个部位过氧化氢酶（CAT）、过氧化物酶（POD）活性和脯氨酸（Pro）含量变化及与其耐寒性的关系.结果表明,低温期间,黄C的中胚轴、胚芽鞘相对生长率显著高于Mo17;黄C的根、中胚轴、胚芽鞘3个部位的CAT、POD活性和Pro含量变化率（其中,中胚轴分别为39.02%、-31.35%和86.86%）均高于Mo17（中胚轴变化率分别为1.86%、-48.67%和12.01%）;黄C 3个部位的质膜透性（中胚轴为22.05%）均低于Mo17（中胚轴为29.11%）.逐步回归分析表明黄C低温期间积累的干物质主要与Pro含量有关,而Mo17主要与POD活性有关,玉米幼苗中胚轴对低温反应最敏感,中胚轴Pro含量变化率与玉米耐寒性关系最密切.     

铅胁迫对黄瓜幼苗抗氧化酶活性及同工酶的影响

采用水培法和聚丙烯酰胺凝胶电泳法,研究铅胁迫对黄瓜幼苗过氧化物酶(POD)、超氧化物歧化酶(SOD)、过氧化氢酶(CAT)活性及同工酶的影响.结果表明,铅胁迫下黄瓜幼苗地上部POD活性除第5天外均随铅浓度的增加而逐渐降低,POD同工酶谱带和表达量减少.在0～500 mg·L^-1铅浓度范围内,SOD活性随铅浓度的增加而增加,第7天达到最大值后急剧下降,低于同期对照值,900 mg·L^-1铅处理SOD活性随时间的延长逐步降低,SOD酶谱带和表达量与铅浓度呈负相关.CAT酶谱带无明显变化,而表达量存在差异.     

Effects of salicylic acid and cold treatments on protein levels and on the activities of antioxidant enzymes in the apoplast of winter wheat leaves

The effects of salicylic acid (SA) and cold on apoplastic protein levels and activities of apoplastic catalase (CAT), peroxidase (POX) and polyphenol oxidase (PPO) were investigated in winter wheat (Triticum aestivum cv. Dogu-88) leaves. The plants were grown with and without 10 microM SA treatment at both control (20/18 degrees C for 30 and 45-day) and cold (10/5 degrees C for 30-day and 5/3 degrees C for 45-day) acclimatisations. Molecular masses of the apoplastic polypeptides were shown ranging in size from 20 to 66 kDa on SDS-PAGE. Accumulation and pattern of the polypeptides were changed by both SA and cold. It is observed that CAT, POX and PPO activities at 45-day control leaves were higher than at 30-day. When the activities with SA and cold treatments are compared to their controls, CAT activities were decreased while POX and PPO activities were increased by both the treatments. When the activities with cold+SA treatment are compared to their cold treatments, CAT and POX activities were decreased while PPO activity was increased by SA. It is concluded that exogenous SA can be involved in cold tolerance by regulating apoplastic proteins and antioxidant enzyme activities.     

Effect of nitric oxide and putrescine on antioxidative responses under NaCl stress in chickpea plants

Chickpea plants were subjected to salt stress for 48 h with 100 mM NaCl, after 50 days of growth. Other batches of plants were simultaneously treated with 0.2 mM sodium nitroprusside (NO donor) or 0.5 mM putrescine (polyamine) to examine their antioxidant effects. Sodium chloride stress adversely affected the relative water content (RWC), electrolyte leakage and lipid peroxidation in leaves. Sodium nitroprusside and putrescine could completely ameliorate the toxic effects of salt stress on electrolyte leakage and lipid peroxidation and partially on RWC. No significant decline in chlorophyll content under salt stress as well as with other treatments was observed. Sodium chloride stress activated the antioxidant defense system by increasing the activities of peroxidase (POX), catalase (CAT) superoxide dismutase (SOD) and ascorbate peroxidase (APX). However no significant effect was observed on glutathione reductase (GR) and dehydro ascorbate reductase (DHAR) activities. Both putrescine and NO had a positive effect on antioxidant enzymes under salt stress. Putrescine was more effective in scavenging superoxide radical as it increased the SOD activity under salt stress whereas nitric oxide was effective in hydrolyzing H₂O₂ by increasing the activities of CAT, POX and APX under salt stress.     

Cell infrastructure and some enzyme activities in rice coleoptiles grown in air and in anoxia

The rice ( Oryza saliva L. cv. S-6) cells in anaerobic coleoptiles maintained their ultra-structure. Most of the organelles did not show significant changes as compared to those from aerobic tissues. However, the number of mitochondria was reduced by 34% and they showed enlarged cristae. Most affected were unspecialized micro-bodies: Their number was reduced by 80% under anaerobiosis and both matrix and membrane structure appeared altered. The activities of the unspecialized microbody enzymes, glycolate oxidase (EC 1.1.3.1), urate oxidase (EC 1.7.3.3) and catalase (EC 1.11.1.6) were alt reduced by anoxia. Catalase decreased to the same extent as the number of microbodies.     

Changes in gas exchange, proline accumulation and antioxidative enzyme activities in three olive cultivars under contrasting water availability regimes

Changes in photosynthetic performance, osmolyte accumulation and the activities of superoxide dismutase (SOD), ascorbate peroxidase (APX), catalase (CAT) and polyphenol oxidase (PPO) were investigated in one-year-old olive cultivars (Chemlali, Meski and Picholine) subjected to contrasting water availability regimes under arid climatic conditions in Tunisia. Shoot elongation rates (SER) and photosynthetic performance were markedly reduced by the water deficit regime (WD) in all cultivars except for Chemlali, which proved to be superior to the other two cultivars with respect to drought tolerance. Higher photosynthetic performance (net photosynthesis (P_n), stomatal conductance (g_s) and transpiration rates (E)) in the Chemlali and Meski cvs. compared to Picholine olive allowed them to maintain better plant water status and shoot elongation rates. Under WD conditions, Chemlali showed a greater capability for proline accumulation. Leaves grown under WD conditions showed signs of oxidative stress such as reduced chlorophyll and carotenoid concentrations. Nevertheless, different cultivars developed certain antioxidative defense mechanisms, including elevated SOD, APX and CAT activities. In contrast, PPO activity decreased under WD circumstances. Comparatively, Chemlali olive displayed better antioxidative enzyme activity, and thus better protection against oxidative stress. These results show that the ability of olive trees to up-regulate the enzymatic antioxidative system might be an important attribute linked to drought tolerance. These findings demonstrate that the association of higher P_n, proline accumulation and antioxidative defenses could be effective in a water-limited environment and may be useful selection criteria in breeding programs with the objective of improving drought tolerance and growth of olive trees, at least under the described environmental conditions.     

Effects of NaCl stress on proline and cation accumulation in salt sensitive and tolerant turfgrasses

Summary Concentrations of proline, sodium and potassium in shoot tissues of five turfgrass species were measured following exposure to 170 mM NaCl salinity stress. Salt tolerant ‘Fults’ alkaligrass and ‘Dawson’ red fescue restricted the accumulation of Na-ions to significatnly low levels compared to the salt sensitive Kentucky bluegrasses (‘Adelphi’ and ‘Ram I’) and ‘Jamestown’ red fescue. Accumulation of proline began in all species within 24 h of initiation of salt stress but at a more rapid rate and higher overall concentration for ‘Fults’ alkaligrass. Proline levels were variable and too low in relation to sodium accumulations to have any significant osmoregulatory role in salt tolerance among all cultivars tested with the possible exception of alkaligrass.     

供氮水平对菠菜营养品质和体内抗氧化酶活性的影响

通过水培实验,研究了供氮水平对菠菜营养品质和抗氧化酶活性的影响.结果表明,供氮水平由4mmol·L^-1增加到8mmol·L^-1,菠菜产量显著增加,叶片中的维生素C(Vc)含量随着供氮浓度由4mmol·L^-1提高到8mmol·L^-1,再提高供氮水平,Vc含量则明显下降.叶片硝酸盐含量随着氮浓度的提高而增加.供氮浓度从4mmol·L^-1增加到8mmol·L^-1,叶片可溶态草酸含量略有下降,再提高供氮水平则明显上升,而草酸总量随供氮水平提高,先显著升高然后略有降低.SOD和POD酶的活性随供氮水平由4mmol·L^-1提高到8mmol·L^-1而增加,再提高供氮水平,酶活性显著下降;CAT活性随供氮水平的增加而降低,叶片MDA含量先降低后显著升高,而游离脯氨酸含量随氮水平的升高而增加.可见供氮水平为8mmol·L^-1时,菠菜叶片具有较高的生物量、Vc含量和抗氧化酶活性,较低的硝酸盐和草酸含量以及较低的MDA和游离脯氨酸含量,表明供氮浓度8mmol·L^-1有利于提高菠菜的产量、营养品质和抗逆能力,是菠菜生长较适宜的供氮水平.     

Salicylic acid mitigates salt induced toxicity through the modifications of biochemical attributes and some key antioxidants in capsicum annuum

Abiotic stress causes extensive loss to agricultural yield production worldwide. Salt stress is one of them crucial factor which leads to decreased the agricultural production through detrimental effect on growth and development of crops. In our study, we examined the effect of a defense growth substance, salicylic acid (SA 1 mM) on mature vegetative (60 Days after sowing) and flowering (80 DAS) stage of Pusa Sadabahar (PS) variety of Capsicum annuum L. plants gown under different concentrations of NaCl (25, 50, 75, 100 and 150 mM) and maintained in identical sets in pots during the whole experiment. Physiological studies indicated that increase in root & shoot length, fresh & dry weight, number of branches per plant, and yield (number of fruits per plant) under salt + SA treatment. Biochemical studies, enzymatic antioxidants like CAT, POX, and non-enzymatic antioxidant such as ascorbic acid (AsA content), carotenoids, phenolics, besides other defense compounds like proline, protein, chlorophyll contents were studied at 10 days after treatment at the mature vegetative and flowering stage. The addition of SA led to lowering of in general, all studied parameters in the mature vegetative stage but increased the same during the flowering stage, especially in the presence of NaCl; although the control I (without SA and NaCl) remained lower in value than control II (with SA, without NaCl). Interestingly, total phenolics were higher in control I (without SA or NaCl) whereas chlorophylls were higher in treatments with SA and NaCl. Thus, physiological concentration of SA (1 mM) appears to be significantly effective against salt stress during the flowering stage. In addition, during the mature vegetative stage, however, proline accumulates in SA treated sets, to help in developing NaCl-induced drought stress tolerance.     

Alterations in the activities of the enzymes of proline metabolism in Ragi (Eleusine coracana) leaves during water stress

Free proline content in Ragi (Eleusine coracana) leaves increased markedly (6 to 85 fold) as the degree of water stress, created by polyethylene gylcol treatment, was prolonged There was also a marginal increase in soluble proteins in the stressed leaves as compared to that in the controls. Water stress stimulated the activities of ornithine aminotransferase and pyrroline-5-carboxylate reductase, the enzymes of proline biosynthesis and markedly inhibited the enzymes involved in proline degradation viz., proline oxidase and pyrroline-5-carboxylate dehydrogenase. These results suggest that increase in free proline content of Ragi leaves could be due to enhanced activities of the enzymes synthesizing proline but more importantly due to severe inhibition of the enzymes degrading proline. These observations establish for the first time, the pathway of proline metabolism in plants by way of detection of the activities of all the enzymes involved and also highlight the role of these enzymes in proline accumulation during water stress.     

Osmotic stress and water stress have opposite effects on putrescine and proline production in excised rice leaves

The effects of water stress and osmotic stress (sorbitol treatment) on the production of putrescine and proline in excised rice leaves were compared. Osmotic stress and water stress were found to affect differentially the levels of putrescine and proline in excised rice leaves. Putrescine accumulation is induced by osmotic stress, whereas proline accumulation is induced by water stress. The effects of ABA on the levels of proline and putrescine are similar to those of water stress, whereas the effects of jasmonic acid methyl ester (JA-Me) are similar to those of osmotic stress. Water stress results in an increase of endogenous ABA is excised rice leaves. However, neither osmotic stress nor JA-Me has effect on endogenous ABA levels in excised rice leaves. Of particular interest is the finding that proline levels increase when putrescine levels induced by osmotic stress or JA-Me are reduced by D-arginine and -methylornithine. L-arginine and L-ornithine applied exogenously also cause an increase in proline levels. It seems that L-arginine and L-ornithine are preferentially utilized as precursors for putrescine accumulation in excised rice leaves treated with osmotic stress and JA-Me, and for proline accumulation in excised rice leaves exposed to water stress and ABA.Abbreviations ABA abscisic acid - BSA bovine serum albumin - ELISA enzyme-linked immunosorbent assay - HPLC high performance chromatography - JA-Me jasmonic acid methyl ester - PVP poly-vinylpyrrolidone     

Effects of water stress on glycolate metabolism in the leaves of rice seedlings (Oryza sativa)

To investigate the effects of water stress on glycolate metabolism, seedlings of a drought-tolerant cultivar (N-22) and a susceptible cultivar (Jaya) of Oryza sativa L. were subjected to water stress for 5, 8 or 10 days. Increasing the duration of water-deficit-stress produced a proportional decrease in relative water content and leaf water potential, reduced glycolate content and catalase (EC 1.11.1.6) activity, but increased glycolate oxidase (EC 1.1.3.1) activity, hydrogen peroxide and glyoxylate contents in the leaves of both cultivars. In a radiotracer experiment, with increasing duration of water stress, the proportion of label increased in 3-phosphoglycerate, glycolate, glycine and serine. The drought-tolerant cultivar (N-22) was affected less than the susceptible cultivar (Jaya). The glycolate pathway metabolism is discussed in relation to photorespiration and the effects of water stress.     

氮钙对香石竹多酚氧化酶、过氧化物酶和叶斑病的影响

1　引　　言近几年安徽省香石竹鲜切花生产得到迅速发展 ,但每年6月至 10月间叶斑病均有较严重发生 ,药物也很难从根本上防止叶斑病的发生和蔓延 ,影响香石竹的生产 .由于多数香石竹生产基地长期只注重用销铵、尿素等Ｎ肥 ,而忽视施用含Ｃａ2 + 肥料和Ｃａ2 + 的生理作用 ,导致栽培基质Ｎ(ＮＯ-3 )、Ｃａ2 + 失去平衡 ,植株对病害抵抗能力降低 .本试验着重研究Ｎ(ＮＯ-3 )、Ｃａ2 + 营养浓度对与抗病性有关的多酚氧化酶、过氧化物酶活性和植株发生叶斑病程度的关系 ,为合理施肥减轻叶斑病对香石竹危害程度提供理论依据 .2　材料与方法2 1　…     

Accumulation of some nitrogen compounds in response to salt stress and their relationships with salt tolerance in rice (Oryza sativa L.) seedlings

The salt-induced accumulation of some nitrogen compounds (free amino acids, ammonium and urea) in shoots of eight rice cultivars differing in salt tolerance was investigated. Salt treatment (100 mM, 6 days) significantly increased the proline content of shoots but this appeared to be a reaction to stress damage and not associated with salt tolerance, because proline contents were higher in the more sensitive cultivars. Besides proline, some other free amino acids also accumulated leading to a significant increase in the total amino acid content of the stressed seedlings. High levels of free ammonium also accumulated under conditions of stress; this was highly correlated with the accumulation of Na⁺ in the shoots and negatively correlated with salt tolerance. The accumulation of ammonium was positively correlated with the accumulation of many free amino acids, and also associated with the production of urea in the stressed seedlings. Results from the present investigations suggest that an increase in the concentration of some free amino acids including proline, may be a result of the reassimilation of the stress-induced ammonium. A high capacity to assimilate ammonium may be an important factor in alleviating the consequence of stress because ammonium can be toxic at high concentrations.     

Silicon-mediated changes of some physiological and enzymatic parameters symptomatic for oxidative stress in spinach and tomato grown in sodic-B toxic soil

We investigated effect of silicon (Si) on the growth, uptake of sodium (Na), chloride (Cl), boron (B), stomatal resistance (SR), lipid peroxidation (MDA), membrane permeability (MP), lipoxygenase (LOX) activity, proline (PRO) accumulation, H₂O₂ accumulation, non-enzymatic antioxidant activity (AA) and the activities of major antioxidant enzymes (superoxide dismutase, SOD; catalase, CAT and ascorbate peroxidase, APX) of spinach and tomato grown in sodic-B toxic soil. Si applied to the sodic-B toxic soil at 2.5 and 5.0 mM concentrations significantly increased the Si concentration in the plant species and counteracted the deleterious effects of high concentrations of Na, Cl and B on root and shoot growth by lowering the accumulation of these elements in the plants. Stomatal resistance, MP, MDA and the concentrations of H₂O₂ and PRO were higher in the plants grown in sodic-B toxic soil without Si: LOX activity of excised leaves of both species was increased by Si. Antioxidant activities of both species were significantly affected by Si, with the activities of SOD, CAT and APX decreased and AA increased by applied Si. For most of the parameters measured, it was found that 5 mM Si was more effective than the 2.5 mM Si. Based on the present work, it can be concluded that Si alleviates sodicity and B toxicity of the plants grown in sodic-B toxic soil by preventing both oxidative membrane damage and also translocation of Na, Cl and B from root to shoots and/or soil to plant, and lowering the phytotoxic effects of Na, Cl and B within plant tissues. It was concluded that tomato was more responsive to Si than spinach since it was more salt sensitive than spinach. To our knowledge, this is the first report that Si improves the combined salt and B tolerance of spinach and tomato grown in naturally sodic-B toxic soil, and which describes membrane-related parameters and antioxidant responses.