Proline In Response Research Articles

Application of ATR-FTIR Spectroscopy for Analysis of Salt Stress in Brussels Sprouts.

Salt stress is one of the environmental stresses that significantly reduces crop productivity and quality worldwide. Methods to overcome salt stress include developing salt-resistant crops by inserting various resistance genes or to diagnosing and responding to the effects of salt stress at an early stage. In this study, we investigate the effects of salinity stress on growth, photosynthetic efficiency, and metabolic changes in Brussels sprouts (Brassica oleracea var. gemmifera). Fresh weight and leaf area decreased significantly with increasing NaCl concentration, indicating that salinity stress has a detrimental effect on plant growth. However, chlorophyll fluorescence parameters did not show significant changes, suggesting that photosynthetic efficiency was not significantly affected over 10 days. Fourier transform infrared (FTIR) spectroscopy revealed notable metabolic adjustments, especially in lipids, plastids, proteins, and carbohydrates, indicating biosynthesis of protective compounds such as anthocyanins and proline in response to salinity stress. Pearson correlation analysis confirmed a strong relationship between NaCl concentration and the observed physiological and metabolic changes. The findings highlight the potential of FTIR spectroscopy as a non-destructive tool for early detection of salinity stress and timely intervention to improve crop resilience and yield. This study highlights the widespread application of FTIR spectroscopy in agricultural research to manage abiotic stresses in crops.

Proline metabolism and its osmotic regulatory role in enhancing hypersalinity tolerance of the razor clam (Sinonovacula constricta)

Proline serves as an important osmolyte in maintaining osmotic pressure and stabilizing cellular structure in aquatic organisms, and is essential for adaptation to salinity stress. This study focuses on the razor clam (Sinonovacula constricta), and investigated the metabolism and synthesis of proline in response to changing salinity. The proline content in gill tissues of razor clams significantly increased over time and with different levels of salinity exposure. The key genes involved in proline metabolism, including (Aminotransferase (ScOAT), Δ1-pyrroline-5-carboxylic acid synthase (ScP5CS), and proline dehydrogenase (ScProDH) were investigated, as well as enzyme activities associated with these genes in razor clams. The expression and activity of P5CS increased significantly under hypersalinity conditions, whereas that of OAT enzyme activity decreased. This suggested that the glutamate pathway had an important role in the proline synthesis during hypersalinity. ProDH activity was negatively correlated with salinity and regulated by the proline content, indicating a significant role of ProDH in proline accumulation under hypersalinity conditions. ScP5CS was heterologously expressed in yeast Saccharomyces cerevisiae, followed by exogenous proline supplementation studies. Recombinant S. cerevisiae incubated with ScP5CS exhibited elevated levels of proline, with the growth performance of recombinant strain surpassing that of the control group after salinity stress. In addition, razor clams exposed to exogenous proline under high salinity conditions showed expedite osmotic regulation, whereby the hemolymph pressure reached equilibrium more quickly. Together, these results indicated that the proline content increased under high salinity, which could improve the tolerance of razor clams to hypersalinity, offering novel perspectives into molecular mechanism of salinity adaptation in shellfish.

Unraveling the evolutionary origin of the P5CS gene: a story of gene fusion and horizontal transfer.

The accumulation of proline in response to the most diverse types of stress is a widespread defense mechanism. In prokaryotes, fungi, and certain unicellular eukaryotes (green algae), the first two reactions of proline biosynthesis occur through two distinct enzymes, γ-glutamyl kinase (GK E.C. 2.7.2.11) and γ-glutamyl phosphate reductase (GPR E.C. 1.2.1.41), encoded by two different genes, ProB and ProA, respectively. Plants, animals, and a few unicellular eukaryotes carry out these reactions through a single bifunctional enzyme, the Δ1-pyrroline-5-carboxylate synthase (P5CS), which has the GK and GPR domains fused. To better understand the origin and diversification of the P5CS gene, we use a robust phylogenetic approach with a broad sampling of the P5CS, ProB and ProA genes, including species from all three domains of life. Our results suggest that the collected P5CS genes have arisen from a single fusion event between the ProA and ProB gene paralogs. A peculiar fusion event occurred in an ancestral eukaryotic lineage and was spread to other lineages through horizontal gene transfer. As for the diversification of this gene family, the phylogeny of the P5CS gene in plants shows that there have been multiple independent processes of duplication and loss of this gene, with the duplications being related to old polyploidy events.

Proline metabolism regulation in Spartina alterniflora and SaP5CS2 gene positively regulates salt stress tolerance in transgenic Arabidopsis thaliana

ABSTRACT It has been discovered that many plants accumulate proline in response to osmotic stress. However, the interaction between salinity stress and proline metabolism-related genes has not been unequivocally identified in Spartina alterniflora. In our research, we successfully cloned key genes: SaP5CS1 and SaP5CS2, SaOAT, SaProT and correlated with expression profiles including SaPDH were also investigated. As the key enzyme genes for proline metabolism, the up-regulated expression of SaP5CS2 played a leading part under salt stress. Chlorophyll contents decreased gradually degree in leaves with prolonged salt stress time. Additionally, transgenic Arabidopsis overexpressing SaP5CS2 showed high tolerance to salt stress and accumulated more proline and H2O2 content was also reduced in salt stress condition. This study provided a foundation for deciphering the molecular osmoregulation underlying proline metabolism in S. alterniflora and SaP5CS2 may play an important role in conferred salinity tolerance by the regulation of proline.

Heterografted chrysanthemums enhance salt stress tolerance by integrating reactive oxygen species, soluble sugar, and proline.

Chrysanthemum, one of the most important commercial ornamental crops, is susceptible to salinity, which limits its cultivation and application in coastal and inland saline areas. Grafting is widely used to improve the salt tolerance of horticultural crops, but the mechanisms of grafted chrysanthemum responses to salt stress remain unclear. In this study, we showed that heterografted chrysanthemums with Artemisia annua as rootstock exhibited increased salt tolerance compared with self-grafted and self-rooted chrysanthemums. Under high salt stress, the roots of heterografted chrysanthemums enrich Na+, resulting in a reduction of Na+ toxicity in the scion, with only a small amount of Na+ being transported to the leaves. On the other hand, the roots of heterografted chrysanthemums alleviated high Na+ stress via enhanced catalase enzyme activity, downregulation of the expression of reactive oxygen species (ROS) accumulation-related genes, massive accumulation of soluble sugars and proline, and upregulation of the expression of heat shock protein-related genes to enhance salt tolerance. In addition, the leaves of heterografted chrysanthemums respond to low Na+ stress by increasing peroxidase enzyme activity and soluble sugar and proline contents, to maintain a healthy state. However, self-grafted and self-rooted plants could not integrate ROS, soluble sugars, and proline in response to salt stress, and thus exhibited a salt-sensitive phenotype. Our research reveals the mechanisms underlying the increased salt tolerance of heterografted chrysanthemums and makes it possible to have large-scale cultivation of chrysanthemums in saline areas.

Differential Impact of Salinity Stress on Seeds Minerals, Storage Proteins, Fatty Acids, and Squalene Composition of New Quinoa Genotype, Grown in Hyper-Arid Desert Environments.

The effects of climate change and soil salinization on dryland ecosystems are already widespread, and ensuring food security is a crucial challenge. In this article, we demonstrate changes in growth performance and seed quality of a new high-yielding quinoa genotype (Q5) exposed to sodium chloride (NaCl), sodium sulfate (Na2SO4), and mixed salts (NaCl + Na2SO4). Differential responses to salt stress in growth performance, seed yield, and seed quality were identified. High salinity (mixed Na2SO4 + NaCl) reduces plant height by ∼30%, shoot and root dry weights by ∼29%, head panicle length and panicle weight by 36–43%, and seed yield by 37%, compared with control conditions. However, the 1,000-seed weight changes insignificantly under salinity. High content of essential minerals, such as Fe, Zn, and Ca in quinoa Q5 seeds produced under salinity, gives the Q5 genotype a remarkable advantage for human consumption. Biomarkers detected in our studies show that the content of most essential amino acids is unchanged under salinity. The content of amino acids Pro, Gly, and Ile positively correlates with Na+ concentration in soil and seeds, whereas the content of squalene and most fatty acids negatively correlates. Variation in squalene content under increasing salinity is most likely due to toxic effects of sodium and chlorine ions as a result of the decrease in membrane permeability for ion movement as a protective reaction to an increase in the sodium ion concentration. Low squalene accumulation might also occur to redirect the NADPH cofactor to enhance the biosynthesis of proline in response to salinity, as both syntheses (squalene and proline) require NADPH. This evidence can potentially be used by the food and pharmaceutical industries in the development of new food and health products.

Effects of water deficit on shoot, root and some physiological characteristics in some greenhouse grown potato cultivars

A greenhouse experiment was conducted to evaluate the response of ten potato cultivars to two levels of soil water in terms of contents of total chlorophyll (Chltot), chlorophyll florescence (Fv/Fm), shoot fresh weight (SFW), shoot dry weight (SDW), root fresh weight (RFW), root dry weight (RDW), mean root length (MRL), root length density (RLD), special root length (SRL), root volume (RV), ascorbate peroxidase (APX), peroxidase (POX) and proline content. Plants were grown under two irrigation levels (30 and 80% depletion of available soil water). Greenhouse test plots were arranged in a randomized complete block design with four replications. Under control level of irrigation, the maximum and the minimum shoot dry weight were achieved for cultivars Satina (12.7 g/plant) and Sante (4.7 g/plant) and root dry weight for Marfona (2.95 g/plant) and Milva (1.17 g/plant), respectively. SDW and RDW were decreased in all cultivars under 80% depletion of available soil water. Based on the biplot analysis cultivars Agria, Satina and Marfona were hence identified as preferable cultivars for 80% depletion of available soil water conditions. However, this investigate has been able to gather evidence that water deficit increased the activities of antioxidant enzymes and proline content in all tested potato cultivars. Also, our results revealed the pivotal role of proline in response of potato plants to water deficit and biomass production.

Proline oxidation fuels mitochondrial respiration during dark-induced leaf senescence in Arabidopsis thaliana.

Leaf senescence is a form of developmentally programmed cell death that allows the remobilization of nutrients and cellular materials from leaves to sink tissues and organs. Among the catabolic reactions that occur upon senescence, little is known about the role of proline catabolism. In this study, the involvement in dark-induced senescence of proline dehydrogenases (ProDHs), which catalyse the first and rate-limiting step of proline oxidation in mitochondria, was investigated using prodh single- and double-mutants with the help of biochemical, proteomic, and metabolomic approaches. The presence of ProDH2 in mitochondria was confirmed by mass spectrometry and immunogold labelling in dark-induced leaves of Arabidopsis. The prodh1 prodh2 mutant exhibited enhanced levels of most tricarboxylic acid cycle intermediates and free amino acids, demonstrating a role of ProDH in mitochondrial metabolism. We also found evidence of the involvement and the importance of ProDH in respiration, with proline as an alternative substrate, and in remobilization of proline during senescence to generate glutamate and energy that can then be exported to sink tissues and organs.

Effect of Arbuscular Mycorrhizal Fungi, Selenium and Biochar on Photosynthetic Pigments and Antioxidant Enzyme Activity Under Arsenic Stress in Mung Bean (Vigna radiata).

Environmental perturbations alter biochemical compounds in food crops. Arsenic (As), a toxic metalloid, is known to affect the cultivation of food crops in many regions of the world; however, the changes in chlorophyll, catalase (CAT), and proline in response to As stress and the role of stress relief substances remain largely unknown in mung bean (Vigna radiate L.). In this study, biochar (BC), arbuscular mycorrhizal fungi (AMF), and selenium (Se) were applied to soils as stress relief substances (under 30 mg kg-1 As stress), and the effects of BC, AMF, and Se on chlorophyll a, chlorophyll b, total chlorophyll, CAT activity, and proline content were studied in different mung bean genotypes. Under As stress, the chlorophyll a, chlorophyll b, and total chlorophyll contents in BARI mung 3, BARI mung 5, and BARI mung 8 were found statistically similar. Meanwhile, CAT activity increased in comparison to the control due to the application of BC, AMF, and Se in mung bean crops. However, proline was found significantly lower in AMF, BC, and Se-treated mung bean. This indicates that oxidative stress was potentially minimized in As-stressed mung bean crops due to the application of these stress relief substances. Notably, AMF was relatively effective against As stress in comparison to BC and Se. It is concluded that BC, AMF, and Se are all highly effective in enhancing antioxidant defenses as well as the nutritional quality of mung bean crops under As stress.

Organic solutes in coconut palm seedlings under water and salt stresses

ABSTRACT The objective of this study was to investigate the biochemical mechanisms associated with isolated and/or concurrent actions of drought and soil salinity in seedlings of coconut tree, through the accumulation of organic solutes (soluble carbohydrates, soluble amino N and free proline) in leaves and roots. The experiment, conducted in a protected environment, in Fortaleza, Brazil, in a randomized block design, in a split-plot arrangement, evaluated the effects of different levels of water stress (plots) by imposing distinct percentages of replacement of water losses through crop potential evapotranspiration - ETpc (20, 40, 60, 80 and 100%), associated with subplots consisting of increasing levels of soil salinity in saturation extract (1.72, 6.25, 25.80 and 40.70 dS m-1) provided by the soils collected in the Irrigated Perimeter of Morada Nova. Salinity did not change the concentration of organic solutes; however, there were increases in leaf and root levels of free proline in response to water stress, which contributes to the osmoregulation and/or osmoprotection of the species under adverse conditions of water supply.

Effects of Salt Stress on Three Ecologically Distinct Plantago Species.

Comparative studies on the responses to salt stress of taxonomically related taxa should help to elucidate relevant mechanisms of stress tolerance in plants. We have applied this strategy to three Plantago species adapted to different natural habitats, P. crassifolia and P. coronopus–both halophytes–and P. major, considered as salt-sensitive since it is never found in natural saline habitats. Growth inhibition measurements in controlled salt treatments indicated, however, that P. major is quite resistant to salt stress, although less than its halophytic congeners. The contents of monovalent ions and specific osmolytes were determined in plant leaves after four-week salt treatments. Salt-treated plants of the three taxa accumulated Na+ and Cl- in response to increasing external NaCl concentrations, to a lesser extent in P. major than in the halophytes; the latter species also showed higher ion contents in the non-stressed plants. In the halophytes, K+ concentration decreased at moderate salinity levels, to increase again under high salt conditions, whereas in P. major K+ contents were reduced only above 400 mM NaCl. Sorbitol contents augmented in all plants, roughly in parallel with increasing salinity, but the relative increments and the absolute values reached did not differ much in the three taxa. On the contrary, a strong (relative) accumulation of proline in response to high salt concentrations (600–800 mM NaCl) was observed in the halophytes, but not in P. major. These results indicate that the responses to salt stress triggered specifically in the halophytes, and therefore the most relevant for tolerance in the genus Plantago are: a higher efficiency in the transport of toxic ions to the leaves, the capacity to use inorganic ions as osmotica, even under low salinity conditions, and the activation, in response to very high salt concentrations, of proline accumulation and K+ transport to the leaves of the plants.

Overexpression of GmProT1 and GmProT2 increases tolerance to drought and salt stresses in transgenic Arabidopsis

The proline transporter protein (ProT) plays an important role in protective stress responses in various plants. However, its function in abiotic stress responses in soybean (Glycine max) remains obscure. In the present study, two soybean ProT genes, namely GmProT1 and GmProT2, were isolated by homologous cloning. GmProT1 and GmProT2 encode polypeptides of 435 and 433 amino acids, respectively. The GmProT1 and GmProT2 proteins showed high similarity to other ProT proteins. GmProT1 and GmProT2 transcripts were detected in different soybean tissues including roots, stems, leaves, flowers, and developmental seeds, and during diverse developmental stages. GmProT1 was strongly expressed in seeds 35 days after flowering. Quantitative real-time PCR analysis showed that the two genes were highly expressed in leaves and could be strongly induced in response to salt and drought conditions and ABA treatment. Transgenic Arabidopsis thaliana plants overexpressing the two genes were generated, which showed that GmProT genes attenuate damage from salt and drought stress. In addition, transgenic Arabidopsis plants accumulated proline in response to salt and osmotic stress. Transcription levels of salinity-responsive gene (RD29B and S0S3) and drought-induced gene (CDPK1) were higher in the transgenic lines than that of wild type plants. Our work provides evidence that GmProT genes function in the response to abiotic stresses and may affect the synthesis and response system of proline.

Differential growth and yield responses of salt-tolerant and susceptible rice cultivars to individual (Na+ and Cl−) and additive stress effects of NaCl

Negative impacts exerted by sodium (Na+) and chloride (Cl−) ions individually as well their possible additive effects (under NaCl) were evaluated on growth and yield reductions in rice, besides investigating whether salt-tolerant genotypes respond differentially than their sensitive counterparts. Though both Na+ and Cl− ions get accumulated in plant tissues under NaCl stress, most research has historically been aimed to decipher harmful effects induced by Na+ ions. Accordingly, physiological and molecular mechanisms involved in Cl− toxicity are not clearly understood in crop plants. To address these issues, 65-day-old plants of two rice cultivars, Panvel-3 (tolerant) and Sahyadri-3 (sensitive) were subjected to Cl−, Na+ and NaCl (each with 100 mM concentration and electrical conductivity of ≈10 dS m−1) stress using soil-based systems. Stress conditions were maintained till harvesting of mature (128-day-old) plants. All three treatments induced substantial antagonistic effects on growth, dry mass, yield components (number of grains per panicle, length, width, thickness and weight of grain, along with the percentage of grains filled) and overall crop yield, with greater impacts under NaCl than its constituent ions. Salinity treatments caused an imbalance in reducing sugars, protein, starch and proline contents, with the greatest magnitude under NaCl. A negative correlation between Cl−/Na+ accumulation and crop yield was witnessed, with higher severity on the sensitive cultivar. The overall magnitude of toxicity was observed highest in NaCl followed by Na+ and Cl−, respectively, suggesting additive effects of constituent ions under NaCl. Both cultivars responded similarly; however, the tolerant cultivar, unlike the sensitive one, kept Na+:K+ ratio <1.0 and accumulated proline in response to salinity treatments used in this study.

Elucidation of salt-tolerance metabolic pathways in contrasting rice genotypes and their segregating progenies.

Differentially expressed antioxidant enzymes, amino acids and proteins in contrasting rice genotypes, and co-location of their genes in the QTLs mapped using bi-parental population, indicated their role in salt tolerance. Soil salinity is a major environmental constraint limiting rice productivity. Salt-tolerant 'CSR27', salt-sensitive 'MI48'and their extreme tolerant and sensitive recombinant inbred line (RIL) progenies were used for the elucidation of salt stress tolerance metabolic pathways. Salt stress-mediated biochemical and molecular changes were analyzed in the two parents along with bulked-tolerant (BT) and bulked-sensitive (BS) extreme RILs. The tolerant parent and BT RILs suffered much lower reduction in the chlorophyll as compared to their sensitive counterparts. Activities of antioxidant enzymes superoxide dismutase (SOD) and peroxidase (POD) and non-enzymatic antioxidant ascorbic acid were much higher in salt-stressed CSR27 and BT RILs than MI48 and BS RILs. Further, the tolerant lines showed significant enhancement in the levels of amino acids methionine and proline in response to salt stress in comparison to the sensitive lines. Similarly, the tolerant genotypes showed minimal reduction in cysteine content whereas sensitive genotypes showed a sharp reduction. Real time PCR analysis confirmed the induction of methionine biosynthetic pathway (MBP) enzymes cystathionine-β synthase (CbS), S-adenosyl methionine synthase (SAMS), S-adenosyl methionine decarboxylase (SAMDC) and serine hydroxymethyl transferase (SHMT) genes in tolerant lines, suggesting potential role of the MBP in conferring salt tolerance in rice variety CSR27. Proteome profiling also confirmed higher expression of SOD, POD and plastidic CbS and other proteins in the tolerant lines, whose genes were co-located in the QTL intervals for salt tolerance mapped in the RIL population. The study signifies integrated biochemical-molecular approach for identifying salt tolerance genes for genetic improvement for stress tolerant rice varieties.

Macrophage migration inhibitory factor (MIF) is rendered enzymatically inactive by myeloperoxidase-derived oxidants but retains its immunomodulatory function

Macrophage migration inhibitory factor (MIF) is an important player in the regulation of the inflammatory response. Elevated plasma MIF is found in sepsis, arthritis, cystic fibrosis and atherosclerosis. Immunomodulatory activities of MIF include the ability to promote survival and recruitment of inflammatory cells and to amplify pro-inflammatory cytokine production. MIF has an unusual nucleophilic N-terminal proline with catalytic tautomerase activity. It remains unclear whether tautomerase activity is required for MIF function, but small molecules that inhibit tautomerase activity also inhibit the pro-inflammatory activities of MIF. A prominent feature of the acute inflammatory response is neutrophil activation and production of reactive oxygen species, including myeloperoxidase (MPO)-derived hypochlorous acid and hypothiocyanous acid. We hypothesized that MPO-derived oxidants would oxidize the N-terminal proline of MIF and alter its biological activity. MIF was exposed to hypochlorous acid and hypothiocyanous acid and the oxidative modifications on MIF were examined by LC-MS/MS. Imine formation and carbamylation was observed on the N-terminal proline in response to MPO-dependent generation of hypochlorous and hypothiocyanous acid, respectively. These modifications led to a complete loss of tautomerase activity. However, modified MIF still increased CXCL-8/IL-8 production by peripheral blood mononuclear cells (PBMCs) and blocked neutrophil apoptosis, indicating that tautomerase activity is not essential for these biological functions. Pre-treatment of MIF with hypochlorous acid protected the protein from covalent modification by the MIF inhibitor 4-iodo-6-phenylpyrimidine (4-IPP). Therefore, oxidant generation at inflammatory sites may protect MIF from inactivation by more disruptive electrophiles, including drugs designed to target the tautomerase activity of MIF.

Salinity induced physiological and biochemical changes in chickpea (Cicer arietinum L.) genotypes

Plant growth and development are adversely affected by salinity- a major environmental stress that limits agricultural production. Chickpea (Cicer arietinum L.) is sensitive to salinity that affects its yield and there is need to identify the tolerant genotypes. The present study was conducted to evaluate the effect of salinity on chickpea genotypes with specific physiological and biochemical attributes contributing to their adaptability to salinity stress. Seven chickpea genotypes both desi (ICC8950, ICCV10, ICC15868, GL26054) and kabuli (BG1053, L550, L552) were evaluated for salinity tolerance. Maximum decrease in relative leaf water content and chlorophyll content was observed with ICC15868 and GL26054 among the desi and L552 from the kabuli genotypes. The photosynthetic pigments, activity of nitrate reductase and relative leaf water content was also reduced in response to salt application with effect being more pronounced in ICC15868, GL26054 and L552 as compared to ICC8950, ICCV10, BG1053 and L550. Lipid peroxidation increases with the increase in NaCl concentration, maximum increment was observed in genotypes ICC15868, GL26054 and L552. Accumulation of proline in response to environmental stresses seems to be widespread among plants. Higher protein fractions were observed with tolerant genotypes in contrast to sensitive genotypes. Salt imposed stress finally caused a higher decline in number of filled pods. On the basis of physiological and biochemical parameters genotypes ICC8950 and ICCV10 from the desi genotypes and BG1053 and L550 from kabuli were identified as the tolerant while ICC15868, GL26054 as the sensitive ones and L552 as the moderately tolerant genotypes. These genotypes could be used as a source of tolerance in breeding programme to develop salt tolerant genotypes.

Biochemical indicators of salt stress in Plantago maritima: Implications for environmental stress assessment

Our study is focused on native spontaneous species of saline ecosystems Plantago maritima. Plants were cultivated at several salt concentrations (0, 50, 100, 200, 300, 400 and 500mM NaCl) in a glass greenhouse under semi-controlled conditions. Growth parameters, water parameters and ionic status were determined and they were used as criteria to assess the response of P. maritima under a salinity gradient. Catalase, guaiacaol and ascobate peroxidase activities, total protein and proline were also determined. Our results show that P. maritima is a facultative halophyte capable of expressing its maximum growth potential at relatively low concentrations of salt (less than 3gl−1 NaCl). At high doses of salt (concentrations>200mM), the decrease in the growth of P. maritima is associated to a decrease in the uptake of K+. There is a disruption of the water intake of their organs and therefore results an invasion of the cytoplasm by Na+ toxic ion. However, stressed plants use K+ more sparingly. They invest especially in the production of biomass expressed by the dry weight of the shoots, and they use Na+ and proline for osmotic adjustment. The halophyte studied is able to accumulate high levels of proline in response to increasing salt concentration. The accumulation of the amino compound, mainly in roots, is interpreted as an indicator of salt tolerance. Additionally, a significant correlation between the tolerance of the plants to salinity and the activity of several antioxidant enzymes has been observed. Hence, we suggest the possibility of using these activities as a biochemical indicator for salt tolerance in P. maritima. Our study points out two types of biomarkers of salt exposure: enzymatic biomarkers in the leaves and proline content in the roots. Both did show very good correlation with salt exposure, and thus may be considered good biomarkers of exposure with a very good dose–response relationship.

Effect of NaCl stress on dihaploid tobacco lines tolerant to Potato virus Y

Salinity is an important abiotic factor that limits plant growth and development. The influence of salt stress induced by sodium chloride on plant growth, proline content, level of lipid peroxidation and activities of antioxidative enzymes was studied in F1 hybrid DH10 and four dihaploid lines (207B, 238C, 239K, 244B) of tobacco (Nicotiana tabacum L.). Dihaploids were obtained from anther-derived haploids of hybrid DH10 and were previously proved to be tolerant to Potato virus Y (PVY). In our study, plants were grown in vitro and exposed to NaCl (100 and 200 mM) for 33 days. All dihaploids and hybrid DH10 showed reduced growth after NaCl treatment. They accumulated significant amounts of sodium and proline in response to salt stress as have already been observed in tobacco and other plant species. In tobacco exposed to NaCl the lipid peroxidation level did not increase and activities of superoxide dismutase (SOD), guaiacol peroxidase (POD), ascorbate peroxidase and catalase (CAT) mostly did not change significantly. The exception was line 239K where salt induced higher activities of SOD, CAT and POD. Two (238C and 244B) out of four dihaploids appeared more susceptible to salt stress as they showed weak growth in correlation with high proline and sodium content. Therefore, it seems that salt tolerance is not associated with tolerance to PVY. Variations in malondialdehyde and proline content as well as in enzymes activities observed among tobacco lines imply that dihaploids have different genetic properties which might result in different sensitivity to NaCl.

Biometric and biochemical attributes of alfalfa seedlings as indicators of stress induced by excessive cadmium

Abstract The biometric and biochemical attributes of alfalfa ( Medicago sativa L.) seedlings were studied after their exposure to 0-120 μM cadmium for 28 days using hydroponic culture. The growth, photosynthetic area and pigment contents (chlorophyll a, b and total) declined significantly ( p ≤ 0.05) in the presence of high cadmium concentrations (90 and 120 μM). A steady increase in lipid peroxidation assessed via MDA production was observed with increasing levels of cadmium. Cadmium uptake by the plant tissues was concentration dependent. The roots accumulated 1020 µg g-1 of cadmium which was two folds than leaves. An enhanced production up to 600 µg g -1 of proline was observed at higher levels of cadmium. Though, cadmium toxicity was expressed in terms of decline in growth variables, chlorophyll content and oxidative damage but restricted transfer of the metal to the aerial tissue and greater production of proline in response to higher metal content seems to alleviate cadmium toxicity. Thus, ability of alfalfa plants to tolerate high cadmium concentrations can be a manifestation of effective defensive mechanism derived from differential accumulation of metal in plant tissues in addition to enhanced production of proline.

Two P5CS genes from common bean exhibiting different tolerance to salt stress in transgenic Arabidopsis

Many plants accumulate proline in response to salt stress. Δ-pyrroline-5-carboxylate synthetase (P5CS) is the rate-limiting enzyme in proline biosynthesis in plants. Plasmid DNA (pCHF3-PvP5CS1 and pCHF3-PvP5CS2) containing the selectable neomycin phosphotransferase gene for kanamycin resistance and Phaseolus vulgaris P5CS (PvP5CS1 and PvP5CS2) cDNA was introduced into Arabidopsis plants using Agrobacterium-mediated gene transfer. Southern blot, northern blot and RT-PCR analyses demonstrated that the foreign genes were integrated into Arabidopsis chromosomal DNA and expressed. Single-gene transformants were analysed in this study. Transgenic plants expressed higher levels of PvP5CS1 and PvP5CS2 transcripts under salt stress conditions than under normal conditions. When treated with 0, 100 and 200 mM NaCl, the average proline content in leaves of transgenic plants was significantly higher (P < 0.01) than control plants. The average relative electrical conductivity (REC) of transgenic lines was significantly lower (P < 0.01) than control plants under salt stress condition. Biomass production of transgenic lines was significantly higher (P < 0.05) than control plants under 200 mM NaCl stress treatment. These results indicated that introducing PvP5CS1 and PvP5CS2 cDNA into transgenic Arabidopsis caused proline overproduction, increasing salt tolerance. Although the expression of PvP5CS1 in L4 lines and PvP5CS2 in S4 lines was the same under salt stress condition, the S4 lines accumulated 1.6 and 1.9 times more proline than the L4 lines under 100 and 200 mM NaCl treatments, respectively. The REC of S4 plants was 0.5 (100 mM NaCl) and 0.6 times (200 mM NaCl) that of L4 plants. The biomass production of S4 plants was 1.6 times (200 mM NaCl) more than in L4 plants. Total P5CS enzyme activity of S4 was significantly higher than that of L4. These results implied that the PvP5CS2 protein had stronger capacity to catalyze proline synthesis than PvP5CS1 under salt stress condition.