Exogenous Pro Research Articles

A Soybean Pyrroline-5-Carboxylate Dehydrogenase GmP5CDH1 Modulates Plant Growth and Proline Sensitivity

Soybean [Glycine max (L.) Merr.], as a globally commercialized crop, is an important source of protein and oil for both humans and livestock. With more frequent extreme weather disasters, abiotic stress has become one of the critical factors restricting soybean production. Proline (Pro) is a well-known substance in plants that responds to abiotic stress. To identify potential effector genes involved in soybean resistance to abiotic stress, we focused on the pyrroline-5-carboxylate dehydrogenase (P5CDH) which is a key enzyme in the degradation process of Pro. Through homologous sequence alignment, phylogenetic tree, and predicted expression, we chose GmP5CDH1 (Glyma.05G029200) for further research. Tissue-specific expression assay showed that GmP5CDH1 had higher expression levels in soybean seed and cotyledon development. Subcellular localization assay revealed that GmP5CDH1 was a nuclear-membrane-localized protein. As the result of the predicted cis-acting regulatory element indicates, the expression level of GmP5CDH1 was induced by low temperature, drought, salt stress, and ABA in soybean. Next, we constructed transgenic Arabidopsis overexpressing GmP5CDH1. The results showed that GmP5CDH1 also strongly responded to exogenous Pro, and overcame the toxicity of abiotic stress on plants by regulating the endogenous concentration of Pro. The interaction between GmP5CDH1 and GmSAM1 was validated through yeast two-hybrid, LUC fluorescence complementary, and BIFC. In conclusion, overexpression of a soybean pyrroline-5-carboxylate dehydrogenase GmP5CDH1 regulates the development of Arabidopsis thaliana by altering proline content dynamically under salt stress, especially improving the growth of plants under exogenous Pro.

Identification of the Key Genes Involved in Proline-Mediated Modification of Cell Wall Components in Rice Seedlings under Trivalent Chromium Exposure.

Chromium (Cr) toxicity exerts a detrimental effect on various physiological, biochemical, and molecular attributes of plants including the structure and functions of cell walls. On the other hand, the exogenous application of proline (Pro) is a beneficial strategy to overcome Cr toxicity. Therefore, it is a novel strategy to find the key genes associated with cell wall composition in rice under trivalent Cr with/without Pro application. A total of 203 genes were activated in the four cell wall biosynthesis pathways under chromium stress, namely cellulose (60), hemicellulose (57), lignin (35), and pectin (51). Based on the expression abundance of microarrays, the number of differentially expressed genes, and the expression level of genes, the lignin pathway was a crucial pathway in response to Cr treatments, followed by the cellulose pathway. Through the estimation of gene expression variation factors between 'Cr' and 'Cr+Pro' treatments, OsUGP1, OsBGLU24, OsBGLU29, OsBGLU33, OsBMY1, and OsBMY2 in the cellulose pathway; OsXTH9, OsXTH10, OsXTH16, OsGAUT3, OsGAUT19, OsGAUT28, OsXTH1, OsGAUT12, and OsGAUT21 in the hemicellulose pathway; OsPAL3, OsPAL3, OsPOX1, and OsPRX77 in the lignin pathway; and OsPME25, OsPGL27, OsPME26, OsPGL9, and OsPLL12 in the pectin pathway are the key genes involved in cell wall modification during Cr exposure with exogenous Pro application. The Pro-mediated activation of these genes could be crucial players in modifying the cell wall structure and composition of rice plants under Cr stress, which needs to be further clarified.

Proline-mediated activation of glyoxalase II improve methylglyoxal detoxification in Oryza sativa L. under chromium injury: Clarification via vector analysis of enzymatic activities and gene expression

Environmental factors affect plants in several ways including the excessive accumulation of methylglyoxal (MG), resulting in dysfunctions of many biological processes. Exogenous proline (Pro) application is one of the successful strategies to increase plant tolerance against various environmental stresses, including chromium (Cr). This study highlights the alleviation role of exogenous Pro on MG detoxification in rice plants induced by Cr(Vl) through modifying the expression of glyoxalase I (Gly I)- and glyoxalase II (Gly II)-related genes. The MG content in rice roots was significantly reduced by Pro application under Cr(VI) stress, however, there was little effect on the MG content in shoots. In this connection, the vector analysis was used to compare the involvement of Gly l and Gly II on MG detoxification in ‘Cr(VI)’ and ‘Pro+Cr(VI)’ treatments. Results exhibited that vector strength in rice roots increased with an increase in Cr concentrations, while there was a negligible difference in the shoots. The comparative analysis demonstrated that the vector strengths in roots under ‘Pro+Cr(VI)’ treatments were higher than ‘Cr(VI)’ treatments, suggesting that Pro improved Gly II activity more efficiently to reduce MG content in roots. Calculation of the gene expression variation factors (GEFs) indicated a positive effect of Pro application on the expression of Gly I and Gly ll-related genes, wherein a stronger impact was in roots than the shoots. Together, the vector analysis and gene expression data reveal that exogenous Pro chiefly improved Gly ll activity in rice roots which subsequently enhanced MG detoxification under Cr(VI) stress.

Exogenous proline activated an integrated response of NER and HR pathways to reduce DNA damage in rice seedlings under chromium stress.

The DNA damage induced by hexavalent chromium [Cr(VI)] pollutant causes a genotoxic effect on rice seedlings. Hereby, we examined the effects of exogenous proline (Pro) on the alleviation of DNA damage in rice seedlings under different effective concentrations of Cr(VI). Our results revealed that Cr(VI) stress induced reactive oxygen species (ROS), i.e., H2O2 and O2·- accumulation in rice seedlings, repressed genes expression activated in the homologous recombination (HR) and nucleotide excision repair (NER) pathways, and caused DNA damage. Exogenous application of Pro increased Cr accumulation in rice roots, but decreased Cr accumulation in rice shoots, wherein Pro application decreased ROS accumulation in both tissues of rice seedlings. The comet assays suggested that exogenous application of Pro significantly alleviated the DNA damage in rice seedlings during Cr(VI) treatments, judged by the Olive tail moment and tail DNA. Transcriptional assays revealed that exogenous Pro upregulated the expression level of genes associated with the HR and NER pathways and triggered coordinated actions of both repairing pathways to modulate DNA lesion in rice plants during exposure to Cr(VI). Calculations from gene expression variation factors showed that regulative effect of exogenous application of Pro on DNA repair pathways was highly activated at 2.0 mg Cr/L. The current study revealed that Cr(VI) affect rice plants and exogenous Pro rescue these effects by the activation of HR and NER pathways.

Proline-mediated modulation on DNA repair pathway in rice seedlings under chromium stress by integrating gene chip and co-expression network analysis.

Chromium (Cr) stress can cause oxidative burst to plants. Application of exogenous proline (Pro) is one of the most effective approaches to improve the tolerance of plants to Cr stress. In this study, we integrated the data of gene chip with co-expression network analysis to identify the key pathways involved in the DNA repair processes in rice seedlings under Cr(VI) stress. Based on KEGG pathway analysis, 158 genes identified are activated in five different types of DNA repair pathways, namely base excision repair (BER, 20 genes), mismatch repair (MMR, 30 genes), nonhomologous end joining (NHEJ, 8 genes), nucleotide excision repair (NER, 56 genes) and homologous recombination (HR, 44 genes). Co-expression network analysis showed that genes activated in DNA repair pathways were categorized into six different modules, wherein Module 1 (45.36%), Module 2 (27.84%) and Module 3 (19.59%) carried more weight than others. Integrating the data of gene chip and co-expression network analysis indicated that coordinated actions of HR and NER pathways are mainly associated with DNA repair processes in Cr(VI)-treated rice seedlings supplied with exogenous Pro. OsCSB, OsXPG, OsBRIP1, OsRAD51C, OsRAD51A2, OsRPA, OsTOPBP1C, OsTOP3, and OsXRCC3 activated in the HR pathway had a stronger impact on repairing DNA damage induced by Cr(VI) stress in rice seedlings supplied with exogenous Pro, while OsXPB1, OsTTDA2, OsTFIIH1, OsXPC, OsRAD23, OsDSS1, and OsRPA located at the NER pathway showed more contribution to repairing DNA damage than others.

Co-application of proline or calcium and humic acid enhances productivity of salt stressed pomegranate by improving nutritional status and osmoregulation mechanisms

Maximizing food production through integrated management of vegetative and root growth is a major challenge to food security and sustainability in the face of population growth, salinity stress conditions and climatic changes specially in arid and semi-arid regions. This study was conducted to evaluate the effect of foliar application with proline (Pro) at 5 mM, calcium (Ca) at 1.5% or control supplemented with soil application of humic acid (Hc) at 0, 15 g/tree on the nutrition status, osmoregulatory mechanisms and productivity of ‘Wonderful’ pomegranate trees growing under salt stress conditions. Soil and foliar treatments were applied three times: at flowering stage (April), 2 months after fruit set (June) and at fruit maturity (August). Individual application of either Hc or Pro or Ca alleviated the adverse effects of salt stress. Moreover, supplemented soil application of Hc with Pro or Ca as foliar application increased significantly leaf Pro, total carbohydrates, N, P, Ca and K contents, as well as K/Na and Ca/Na ratio. While it significantly decreased leaf Na and Cl concentration. Furthermore, supplemented application of Hc resulted in the highest decrease in leaf Na and Cl concentrations by 94.59%, 44.79% when combined with Pro and by 51.35%, 31.28%, when combined with Ca. In addition, Hc treatment led to the highest mean fruit yield by 139.56% and 90.73%, respectively as mean of both seasons for Pro and Ca treatments, respectively. The results suggest that, exogenous Pro and Ca supplemented with Hc can mitigate salt stress in ‘Wonderful’ pomegranate through enhancing osmoprotectants accumulaton.

Proline-mediated regulation on jasmonate signals repressed anthocyanin accumulation through the MYB-bHLH-WDR complex in rice under chromium exposure.

Toxic metal-induced overaccumulation of anthocyanin (ATH) in plants can oxidize proteins and break DNA. Herein, the role of exogenous proline (Pro) on the repression of ATH accumulation in rice seedlings during hexavalent chromium [Cr(VI)] exposure was studied. Results indicated that exogenous Pro-mediated regulation of jasmonate signals activated the MYB-bHLH-WDR complex to repress ATH accumulation in rice tissues under Cr(VI) stress. Biochemical and transcript analysis indicated that exogenous Pro promoted the synthesis of jasmonic acid (JA) and its molecularly active metabolite jasmonic acid isoleucine (JA-Ile) in rice tissues under Cr(VI) stress. Increment in the endogenous level of jasmonates positively triggered the expression of genes responsible for the JA signaling pathway and activated the MYB-bHLH-WDR complex, eventually repressing the glycosylation of anthocyanidin to form ATH in rice tissues. In conclusion, exogenous proline-mediated regulation on jasmonate signals was tissue-specific under Cr(VI) stress and a more positive effect was detected in shoots rather than roots.

Mathematical quantification of interactive complexity of transcription factors involved in proline-mediated regulative strategies in Oryza sativa under chromium stress

Involvement of transcription factor (TFs) in governing genes at transcription or post transcription level is known to have affirmative impact on plant physiological and morphological development, especially during environmental abuse. Application of exogenous proline (Pro) is one among the effective approaches to strengthen plant resistance against stresses. However, Pro-mediated regulative strategies of TFs in responses to the chromium (Cr) in rice plants through the gene interaction network are still not clear. In the current study, Pro-mediated interactive complexity of various TFs (i.e., MYB, NAC, WRKY, bHLH, and bZIP) under hexavalent chromium [Cr(VI)] was investigated using Agilent 4 × 44 K rice gene chip and gene interactive probability model (GIPM). Results showed that exogenous Pro had a negligible effect on Cr uptake in rice plants, while a small positive response in biomass accretion of rice seedlings was observed under Cr(VI)+Pro treatments which was to certain extend greater than single Cr(VI) treatments. Rice microarray analysis showed that Cr(VI) significantly (p < 0.05) repressed the expression of TFs in the rice roots and shoots, while the application of exogenous Pro significantly (p < 0.05) up-regulated the expression levels of some TFs in rice tissues. Mathematical modularization indicated that Pro-mediated interaction between MYB and NAC carried more weightage than other TFs in rice roots and shoots under Cr(VI) stress. Overall, our study provides convincing evidence to confirm a positive role of exogenous Pro on reducing the negative impact exerted by Cr(VI) on rice plants through regulating expression and interaction of different TFs.

中华蚊母树在干旱-水淹交叉胁迫下形态和活性氧代谢的适应机制

PDF HTML阅读 XML下载 导出引用 引用提醒 中华蚊母树在干旱-水淹交叉胁迫下形态和活性氧代谢的适应机制 DOI: 10.5846/stxb202008062050 作者: 作者单位: 作者简介: 通讯作者: 中图分类号: 基金项目: 国家自然科学基金项目（51779127）；国家111引智基地项目（D20015）；中国科学院水生植物与流域生态重点实验室开放课题基金（E0520204）；三峡库区珍稀植物种子保存技术与设施研究科研项目-耐水淹种质资源调查及收集（SDHZ2021346） Adaptive mechanism of morphology and reactive oxygen species metabolism of Distylium chinense seedlings to alternate drought and submergence stresses Author: Affiliation: Fund Project: 摘要 | 图/表 | 访问统计 | 参考文献 | 相似文献 | 引证文献 | 资源附件 | 文章评论 摘要:为阐明中华蚊母树（Distylium chinense）在消落带干旱-水淹交叉胁迫下的形态和活性氧（ROS）代谢适应机制，通过控制实验模拟了三峡水库消落带的水文节律，研究了干旱-水淹交叉胁迫及恢复过程施加不同外源物质对中华蚊母树形态学和ROS清除的变化。结果表明：（1）前期干旱胁迫增强了中华蚊母树对后期水淹胁迫的适应，主要表现在叶片脱落、大量不定根的形成及茎基部膨大等形态学的变化；（2）干旱或水淹单一胁迫下，中华蚊母树·OH、O2·等ROS水平明显高于对照，表现出氧化应激反应，其超氧化物歧化酶（SOD）、过氧化氢酶（CAT）、抗坏血酸过氧化物酶（APX）等抗氧化系统酶活性及脯氨酸（Pro）等抗氧化系统小分子含量也均显著高于对照，表现出一定的抗氧化防御作用机制，且在复合胁迫下，SOD、CAT、APX酶活性及Pro含量显著高于单一胁迫；（3）恢复阶段，相关性分析表明，中华蚊母树清除ROS （·OH、O2·）的酶促（SOD、CAT、APX）及非酶促（Pro）系统具有一定的协同性。同时，恢复阶段施加脱落酸（ABA），内源Pro显著高于正常水平；施加Pro，SOD、CAT等抗氧化酶活性显著高于对照；施加可溶性糖（Glu），APX活性显著升高。表明外源添加ABA、Pro、Glu，均对ROS的清除产生显著影响，同时，SOD活性显著高于CAT和APX活性，因此SOD可能是中华蚊母树清除冗余ROS的主要因子。因此，中华蚊母树形态学的变化、较高的脯氨酸积累和对ROS有效的清除防御，可能是其适应干旱-水淹交叉胁迫的主要机制，ABA、Pro、Glu等外源物质的添加对中华蚊母树的胁迫恢复过程具有显著的促进作用，前期干旱可提高中华蚊母树植株对后期水淹的耐受能力。 Abstract:Distylium chinense, the genus Distylium of the Hamamelidacea, is an evergreen perennial shrub between 0.8 and 1.2 m in height, native to the riparian areas and wetlands and a dominant species in the riparian areas of the Three Gorges Reservoir (TGR) region of the Yangtze River and its branches. Our previous studies showed that D. chinense had a certain survival rate in the middle and upper part of the water level fluctuation zone (WLFZ) of the TGR to alternate drought and submergence stresses, but the adaptive mechanisms of its morphology and reactive oxygen species (ROS) metabolism is still unknown. To evaluate its adaptation mechanisms of D. chinense to alternate drought and submergence stresses, a simulation alternate drought and submergence experiment was conducted, and morphology and ROS metabolism of D. chinense seedlings and their recovery growth with different exogenous substances, i.e., abscisic acid(ABA), proline(Pro) and glucose (Glu), were analyzed, including adventitious roots, plant height, base stem diameter, leaf numbers, ROS content, enzymatic and non-enzymatic antioxidant parameters. The results were as follows. (1) The early drought stress stimulated the adaptation of D. chinense seedlings to the later flooding stress.The adaptation mainly refered as more morphological changes of stems and roots i.e., stem base hypertrophy, formation and development of adventitious roots. (2) Comparing to the control group, there were significant increases in the ·OH and the O2· in D. chinense seedlings under either drought or flooding stress, which showed oxidative stress responses occurred and their superoxide dismutase (SOD), ascorbate peroxidase (APX), and catalase (CAT) activities and the proline (Pro) contents were also significantly higher than those of controls, which showed that the enzymatic and non-enzymatic antioxidants were induced as a defense mechanism. Under the alternate drought and submergence stresses, the activities of SOD, APX and CAT and Pro contents were higher than those under single drought or flooding stress. (3) Correlation analysis showed that to a certain extent there were coordinated effects between the enzymatic (SOD, CAT, APX) and non-enzymatic (Pro) systems in ROS-scavenging in D. chinense. Exogenous ABA significantly triggered an increase of endogenous Pro contents compared to the controls during the recovery growth and the activity of SOD, CAT and APX was significantly higher than those of the control group under exogenous Pro treatment. Of these, the activities of SOD were significantly higher than those of CAT and APX. The activities of APX increased significantly when soluble sugar (Glu) was added. The ABA, Pro and Glu all had significant influences on the removal of ROS and the ability of D. chinense to up-regulate the SOD activity might be an important attribute for the removal of redundant ROS linked to drought and flooding tolerance. Thus the higher Pro accumulation and effective defense to ROS could be the main physiological adaptation mechanisms for D. chinense under the alternate drought and submergence stresses. The combination of morphological changes and the up-regulating of enzymatic and non-enzymatic anti-oxidative system might be the main mechanisms of D. chinense adapting to alternate drought and flooding stresses. The ABA, Pro and Glu treatment significantly promoted the recovery growth of D. chinense.. The drought in the early stage could improve the tolerance to submergence in the later stage. 参考文献 相似文献 引证文献

Discovery of Prolyl-tRNA Synthetase As a Novel Target in Multiple Myeloma

Discovery of Prolyl-tRNA Synthetase As a Novel Target in Multiple Myeloma

Regulation of Reactive Oxygen Species Metabolism and Glyoxalase Systems by Exogenous Osmolytes Confers Thermotolerance in Brassica napus

In the current study, the beneficial role of proline (Pro) and glycinebetaine (GB) in alleviating high temperature (HT) stress was investigated in Brassica napus L. (rapeseed) seedlings. Ten-d-old rapeseed seedlings were treated with and/or without 2 mM Pro and 2 mM GB and exposed to 38/28 °C day/night (16 and 8 h) temperature for 24 and 48 h. Heat stress induced high amount of hydrogen peroxide (H2O2), lipoxygenase (LOX) and malondialdehyde (MDA) level. Exogenous Pro and GB addition in HT-affected plants increased the levels of ascorbate (AsA) and reduced glutathione (GSH) and their redox pool. Exogenous Pro and GB increased functions of AsA-GSH pathways enzymes along with catalase (CAT), glutathione S-transferase (GST) and glutathione peroxidase (GPX) in HT exposed plants. The enhanced antioxidant defense system by supplementation of Pro and GB helped to reduce oxidative stress and photosynthetic pigments damage caused by HT-induced stress. High methylglyoxal (MG) content decreased upon exogenous Pro and GB application along with enhanced activities of glyoxalase I (Gly I) and Gly II enzymes in HT-stressed plants. Applied Pro and GB under HT stress enhanced endogenous Pro level further to prevent excess water loss and also improved relative water content. Thus, Pro- and/or GB-induced regulatory interactions between ROS and MG detoxification systems may be a useful approach for the reversal of HT-induced oxidative stress.

Olive trees response to lead stress: Exogenous proline provided better tolerance than glycine betaine

Glycine betaine (GB) and proline (Pro) function as compatible solutes and are upregulated in plants under abiotic stresses. The objective of this study was to investigate whether exogenous GB and Pro could improve lead (Pb) tolerance in young olive trees (Olea europaea L.). Comparison between the effect of GB and Pro on Pb-stressed olive trees was realized. Two-year-old olive trees were subjected for five months to two lead-stress levels (150 and 450 mg Pb (NO3)2 kg−1 soil). GB and Pro were supplied through the irrigation water at 20 mM concentration. In both root and leaf tissues, an increase in electrolyte leakage and in oxidative stress markers, such as hydrogen peroxide and thiobarbituric acid reactive substances was observed despite the elevation of antioxidant enzymes activities as well as non-enzymatic antioxidants. Interestingly, GB and Pro supplementation mitigated the adverse Pb effects on O. europaea trees. Indeed, they reduced Pb content and increased the enzymatic and non-enzymatic antioxidants parameters. Thus, oxidative damage was reduced and better levels of plant biomass were obtained. The exogenous Pro appeared to be a better ameliorator than the GB in protecting young O. europaea trees against Pb toxicity.

Exogenous proline stimulates type I collagen and HIF-1\u03b1 expression and the process is attenuated by glutamine in human skin fibroblasts

Abundance of proline (Pro) in collagen molecule led us to investigate whether Pro supply affects collagen biosynthesis in human skin fibroblasts. Treatment of the cells with milimolar concentrations (5 and 10 mM) of Pro for 24 and 48 h contributed to increase in α1 subunit of collagen type I (COL1A1) expression in both cells and culture medium. However, the effect was more pronounced in glutamine-free medium. In such condition, Pro induced collagen expression by about twofold in the cells, while in the medium only by about 30% during 24 h incubation, compared to control. In the presence of glutamine (Gln), exogenous Pro stimulated intracellular collagen expression only by about 30% during 24 h of fibroblasts incubation, and it was not accompanied by adequate increase of collagen secretion into medium. Gln alone stimulated the processes by about 2–3 fold during the course of the experiment. Pro-dependent increase in collagen expression in Gln-free medium was accompanied by increase in prolidase activity and expression of pAkt. In both Gln-free medium and Gln-supplemented medium, Pro induced expression of p53 and HIF-1α. The data suggest that availability of Gln, as a substrate for Pro biosynthesis, determine the utilization of exogenous Pro for the collagen biosynthesis.

Arabidopsis Proline Dehydrogenase Contributes to Flagellin-Mediated PAMP-Triggered Immunity by Affecting RBOHD

Plants activate different defense systems to counteract the attack of microbial pathogens. Among them, the recognition of conserved microbial- or pathogen-associated molecular patterns (MAMPs or PAMPs) by pattern-recognition receptors stimulates MAMP- or PAMP-triggered immunity (PTI). In recent years, the elicitors, receptors, and signaling pathways leading to PTI have been extensively studied. However, the contribution of organelles to this program deserves further characterization. Here, we studied how processes altering the mitochondrial electron transport chain (mETC) influence PTI establishment. With particular emphasis, we evaluated the effect of proline dehydrogenase (ProDH), an enzyme that can load electrons into the mETC and regulate the cellular redox state. We found that mETC uncouplers (antimycin or rotenone) and manganese superoxide dismutase deficiency impair flg22-induced responses such as accumulation of reactive oxygen species (ROS) and bacterial growth limitation. ProDH mutants also reduce these defenses, decreasing callose deposition as well. Using ProDH inhibitors and ProDH inducers (exogenous Pro treatment), we showed that this enzyme modulates the generation of ROS by the plasma membrane respiratory burst NADPH oxidase homolog D. In this way, we contribute to the understanding of mitochondrial activities influencing early and late PTI responses and the coordination of the redox-associated mitochondrial enzyme ProDH with defense events initiated at the plasma membrane.

P5CDH affects the pathways contributing to Pro synthesis after ProDH activation by biotic and abiotic stress conditions.

Plants facing adverse conditions usually alter proline (Pro) metabolism, generating changes that help restore the cellular homeostasis. These organisms synthesize Pro from glutamate (Glu) or ornithine (Orn) by two-step reactions that share Δ1 pyrroline-5-carboxylate (P5C) as intermediate. In the catabolic process, Pro is converted back to Glu using a different pathway that involves Pro dehydrogenase (ProDH), P5C dehydrogenase (P5CDH), and P5C as intermediate. Little is known about the coordination of the catabolic and biosynthetic routes under stress. To address this issue, we analyzed how P5CDH affects the activation of Pro synthesis, in Arabidopsis tissues that increase ProDH activity by transient exposure to exogenous Pro, or infection with Pseudomonas syringae pv. tomato. Wild-type (Col-0) and p5cdh mutant plants subjected to these treatments were used to monitor the Pro, Glu, and Orn levels, as well as the expression of genes from Pro metabolism. Col-0 and p5cdh tissues consecutively activated ProDH and Pro biosynthetic genes under both conditions. However, they manifested a different coordination between these routes. When external Pro supply was interrupted, wild-type leaves degraded Pro to basal levels at which point Pro synthesis, mainly via Glu, became activated. Under the same condition, p5cdh leaves sustained ProDH induction without reducing the Pro content but rather increasing it, apparently by stimulating the Orn pathway. In response to pathogen infection, both genotypes showed similar trends. While Col-0 plants seemed to induce both Pro biosynthetic routes, p5cdh mutant plants may primarily activate the Orn route. Our study contributes to the functional characterization of P5CDH in biotic and abiotic stress conditions, by revealing its capacity to modulate the fate of P5C, and prevalence of Orn or Glu as Pro precursors in tissues that initially consumed Pro.

Exogenous proline application ameliorates toxic effects of arsenate in Solanum melongena L. seedlings

Hydroponic experiments were conducted to investigate an effect of exogenous application of proline (Pro; 25µM) in alleviating arsenate (AsV; 5 and 25µM) toxicity in Solanum melongena L. (eggplant) seedlings. Exposure of AsV declined growth of eggplant, which was coincided with an enhanced accumulation of As. However, exogenous Pro application alleviated AsV toxicity in eggplant seedlings by reducing the accumulation of As. The fluorescence characteristics (JIP-test): φP0, Ψ0, φE0, PIABS, ABS/RC, TR0/RC, ET0/RC, DI0/RC, NPQ and qP were also affected by AsV. However, the effects of AsV were more prominent on PIABS DI0/RC and NPQ. In Pro treated seedlings, following parameters viz. φP0, Ψ0, φE0 and PIABS were stimulated, while, energy flux parameters (ABS/RC, TR0/RC, ET0/RC and DI0/RC) were inhibited. Toxic effects of AsV on photochemistry of photosystem II (PS II) were ameliorated by an exogenous application of Pro. Oxidative stress markers: superoxide radical, hydrogen peroxide and malondialdehyde (lipid peroxidation) were enhanced by AsV exposure, however, their levels were significantly diminished by an exogenous application of Pro. Treatment of AsV stimulated the activities of superoxide dismutase, peroxidase and catalase except that of glutathione-S-transferase. Exogenous Pro application improved the activities of enzymatic antioxidants. The level of endogenous Pro was higher in AsV treated as well as in Pro fed seedlings. The activity of a key enzyme of Pro biosynthesis: Δ1-pyrroline-5-carboxylate synthetase was higher in Pro fed seedlings. The activity of Pro dehydrogenase was inhibited under AsV stress, and its activity was minimum in case of Pro+AsV combination. These results indicate that Pro metabolism could play a key role in regulating the accumulation of As and levels of antioxidants, which concomitantly result into a better growth of eggplant seedlings when compared to the AsV treatments alone.

Exogenous proline and glycine betaine mediated upregulation of antioxidant defense and glyoxalase systems provides better protection against salt-induced oxidative stress in two rice (Oryza sativa L.) varieties.

The present study investigates the roles of exogenous proline (Pro, 5 mM) and glycine betaine (GB, 5 mM) in improving salt stress tolerance in salt sensitive (BRRI dhan49) and salt tolerant (BRRI dhan54) rice (Oryza sativa L.) varieties. Salt stresses (150 and 300 mM NaCl for 48 h) significantly reduced leaf relative water (RWC) and chlorophyll (chl) content and increased endogenous Pro and increased lipid peroxidation and H2O2 levels. Ascorbate (AsA), glutathione (GSH) and GSH/GSSG, ascorbate peroxidae (APX), monodehydroascorbate reductase (MDHAR), dehydroascorbate reductase (DHAR), glutathione reductase (GR), glutathione peroxidase (GPX), catalase (CAT), and glyoxalase I (Gly I) activities were reduced in sensitive variety and these were increased in tolerant variety due to salt stress. The glyoxalase II (Gly II), glutathione S-transferase (GST), and superoxide dismutase (SOD) activities were increased in both cultivars by salt stress. Exogenous Pro and GB application with salt stress improved physiological parameters and reduced oxidative damage in both cultivars where BRRI dhan54 showed better tolerance. The result suggests that exogenous application of Pro and GB increased rice seedlings' tolerance to salt-induced oxidative damage by upregulating their antioxidant defense system where these protectants rendered better performance to BRRI dhan54 and Pro can be considered as better protectant than GB.

Effects of exogenous proline and trehalose on physiological responses in rice seedlings during salt-stress and after recovery

Two osmoprotectants (proline &amp;ndash; Pro; trehalose &amp;ndash; Tre) were exogenously supplied to seedlings of rice cvs. Pokkali (PK &amp;ndash; salt-tolerant) and Khao Dawk Mali 105 (KDML105 &amp;ndash; salt-sensitive) to investigate their effects on plants exposed to 200 mmol/L NaCl for 6 days and 5 days after recovery from stress. The reduction of growth, increase in Na&lt;sup&gt;+&lt;/sup&gt; to K&lt;sup&gt;+&lt;/sup&gt; ratio, high level of hydrogen peroxide (H&lt;sub&gt;2&lt;/sub&gt;O&lt;sub&gt;2&lt;/sub&gt;) content, enhanced activity of antioxidant enzymes (superoxide dismutase &amp;ndash; SOD, peroxidase &amp;ndash; POX, catalase &amp;ndash; CAT and ascorbate peroxidase &amp;ndash; APX) were observed in both rice cultivars under NaCl treatments. Exogenous Pro and Tre supplement to NaCl-stressed plants did not mitigate the reduction of growth during salt-stress. Nevertheless, during recovery plants previously supplied with Tre showed markedly higher percentage of growth recovery than those treated with NaCl alone or supplied with Pro. The beneficial effect of Tre on growth recovery was clearly demonstrated in KDML105 in which growth enhancement was related to reduction in Na&lt;sup&gt;+&lt;/sup&gt; to K&lt;sup&gt;+&lt;/sup&gt; ratio. Exogenous Pro was able to reduce H&lt;sub&gt;2&lt;/sub&gt;O&lt;sub&gt;2&lt;/sub&gt; in both cultivars during salt stress whereas Tre could reduce it only in KDML105. Exogenous Tre did not enhance any antioxidant enzymes during stress but enhanced APX activity in KDML105 during recovery. Exogenous Pro enhanced the activity of APX in PK, and POX, CAT and APX in KDML105 during both stress and recovery period. &amp;nbsp; &amp;nbsp;

Exogenous proline and trehalose promote recovery of rice seedlings from salt-stress and differentially modulate antioxidant enzymes and expression of related genes

Proline (Pro) and Trehalose (Tre) function as compatible solutes and are upregulated in plants under abiotic stress. They play an osmoprotective role in physiological responses, enabling the plants to better tolerate the adverse effects of abiotic stress. We investigated the effect of exogenous Pro and Tre (10mM) in seedlings of Thai aromatic rice (cv. KDML105; salt-sensitive) during salt stress and subsequent recovery. Salt stress (S, NaCl) resulted in growth reduction, increase in the Na+/K+ ratio, increase in Pro level and up-regulation of Pro synthesis genes (pyrroline-5-carboxylatesynthetase, P5CS; pyrroline-5-carboxylate reductase, P5CR) as well as accumulation of hydrogen peroxide (H2O2), increased activity of antioxidative enzymes (superoxide dismutase, SOD; peroxidase, POX; ascorbate peroxidase, APX; catalase, CAT) and transcript up-regulation of genes encoding antioxidant enzymes (Cu/ZnSOD, MnSOD, CytAPX, CatC). Under salt stress, exogenous Pro (PS; Pro+NaCl) reduced the Na+/K+ ratio, further increased endogenous Pro and transcript levels of P5CS and P5CR, but decreased the activity of the four antioxidant enzymes. The transcription of genes encoding several antioxidant enzymes was upregulated. Exogenous Tre (TS; Tre+NaCl) also reduced the Na+/K+ ratio and strongly decreased endogenous Pro. Transcription of P5CS and P5CR was upregulated, the activities of SOD and POX decreased, the activity of APX increased and the transcription of all antioxidant enzyme genes upregulated. Although exogenous osmoprotectants did not alleviate growth inhibition during salt stress, they exhibited a pronounced beneficial effect during recovery period showing higher percentage of growth recovery in PS (162.38%) and TS (98.43%) compared with S (3.68%). During recovery, plants treated with PS showed a much greater reduction in endogenous Pro than NaCl-treated (S) or Tre-treated plants (TS). Increase in CAT activity was most related to significant reduction in H2O2, particularly in the case of PS-treated plants. Advantageous effects of Pro were also associated with increase in APX activity during recovery.

Effects of Exogenous Pro on Resistance of Trapella sinensis Olive to Cd2+ Stress

以高等水生植物茶菱（Trapella sinensis Olive）为材料,研究了外施不同浓度的脯氨酸（Pro）对Cd^2＋毒害的缓解效应.结果表明,茶菱体内Pro含量在单一Cd^2＋毒害下明显增加,外施Pro后,显著降低;叶绿素和可溶性蛋白含量在单一Cd^2＋毒害下大幅下降,外施Pro后,失绿症状有所减轻;保护酶（sOD、CAT、POD）系统在单一Cd^2＋毒害下比例失调,平衡被打破,活性下降,外施Pro后,SOD、POD活性较对照升高,CAT持平,其比例维持在正常范围内;O-·2产生速率在单一Cd2＋毒害下大大加快,外施Pro后,其产生速率恢复正常水平;丙二醛（MDA）含量在单一Cd2＋毒害下积累加剧,外施Pro后,积累程度有所降低.因此,外源Pro可以通过减轻叶绿素和可溶性蛋白的降解,促进蛋白合成,减缓MDA积累,降低O-·2产生速率,增强保护酶活性来增强植物对重金属胁迫的抗性,增强植物对逆境的适应能力.本实验中Pro作用的最佳浓度范围为40～60mg·L-1.