Free proline accumulation in drought-stressed plants

Summary Free-proline accumulation was measured in leaves of intact wheat (Triticum vulgare L. cv. Kalyan Sona), plantago (Plantago ovata Forsk-Isabgool), papavar (Papaver somnifera L. Opium poppy) and mustard (Brassica juncea L. var. Varuna) grown in the field with low to high field water content and thus they were subjected to water stress. Leaf water deficit in percentage was used to determine the degree of stress at the time of proline anlysis.Free proline content was higher in mustard leaves as compared to wheat, plantago and papavar leaves. Water stress enhances the proline content but at same water deficit level the content differ in the leaves of the plants studied.     

Metabolic implications of stress-induced proline accumulation in plants

In many plants, free proline accumulates in response to the imposition of a wide range of biotic and abiotic stresses. Controversy has surrounded the extent to which this shift in nitrogen metabolism benefits plants under adverse environmental conditions. Most attempts to account for the phenomenon have focused on the ability of proline to mediate osmotic adjustment, stabilise subcellular structures and scavenge free radicals. However, often the cytoplasmic pool of free proline even after the imposition of stress is insufficient size to account for pronounced biophysical effects.Alternatively, selective preservation of this stress-induced response may relate to endpoints other than simply augmenting the cellular pool of free proline. Proline accumulation may reduce stress-induced cellular acidification or prime oxidative respiration to provide energy needed for recovery. High levels of proline synthesis during stress may maintain NAD(P)+/NAD(P)H ratios at values compatible with metabolism under normal conditions. Consideration of the cofactor preference of plant 1-pyrroline-5-carboxylate (P5C) reductase as well as the in vivo concentrations of the two pyridine nucleotide cofactors and their respective redox ratios suggests that even a small increase in proline biosynthesis might have a large impact on the level of reduction of the cellular NADP pool. The increased NADP+/NADPH ratio mediated by proline biosynthesis is likely to enhance activity of the oxidative pentose phosphate pathway. This would provide precursors to support the demand for increased secondary metabolite production during stress as well as nucleotide synthesis accompanying the accelerated rate of cell division upon relief from stress, when oxidation of proline is likely to provide an important energy source for ADP phosphorylation. Thus, the extreme sensitivity of the metabolic processes of proline synthesis and degradation themselves may be of benefit by regulating metabolic processes adversely affected by stress. This viewpoint is supported by consideration of other physiological phenomena not directly related to stress responses, but in which proline metabolism may also play a regulatory role.A mechanism is proposed whereby the interconversions of proline and P5C in different cell types and the associated transfer of redox potential between tissues may constitute a form of metabolic signalling within higher plants. Stress-related alterations in proline metabolism may impinge on systems of redox control of plant gene expression.     

Free proline accumulation in drought-stressed plants under laboratory conditions

Summary Free proline accumulation was measured in leaves of intact sorghum (Sorghum bicolor L. cv. Pioneer 846) and soybean (Glycine max. L. cv. Calland) grown in growth chambers and subjected to ‘normal’ drought stress. Stomatal diffusive resistance and leaf water potential were used to determine the degree of stress at the time of proline analysis. Free proline did not accumulate markedly in either species until each was severely stressed, indicating that proline is not a sensitive indicator of drought stress. Free proline accumulated under less stress in soybean than in sorghum. Since soybean is less drought resistant than sorghum, proline accumulation may be an indicator of drought resistance or susceptibility. re]19731003     

Arsenic accumulation of common plants from contaminated soils

A pot experiment was conducted to investigate the relationship between soluble concentrations of arsenic (As) in soil and its accumulation by maize (Zea mays), English ryegrass (Lolium perenne), rape (Brassica napus) and sunflower (Helianthus annuus) on two different soils: a calcareous Regosol (silty loam) and a non-calcareous Regosol (sandy loam). Arsenic (Na₂HAsO₄·7H₂O) was applied to obtain comparable soluble As concentrations in the two soils. In both soils, soluble As concentrations, extracted with 0.1 M NaNO₃, were found to correlate better with As concentrations in plants after 4 month of growth than total soil concentrations, extracted with 2 M HNO₃. With all four plant species, the relationship between the soluble As concentration in the soil and As that in the plants was non- linear, following Michaelis-Menten kinetics. Similar soluble As concentrations in the two soils did not result in a similar As concentration in the plants. Except for maize, arsenic transport from roots to shoots was significant, resulting in As concentrations in the leaves and grains above the Swiss tolerance limits for fodder and food crops (4 and 0.2 mg As kg^–1, respectively). Based on these results we suggest that beside As solubility, P availability and P demand, which are plant specific, have to be taken into account to predict the uptake of As by crop plants from As contaminated soils and to predict the risk of arsenic entering into the food chain.     

Heavy metal accumulation in plants grown in heavily polluted soils

In a polluted area, samples of soils and plants were collected and analyzed in order to assess the mobility and accumulation of Cd, Pb and Zn in plants as influenced by the of different metal forms in soil. Metals were accumulated in roots. Metal mobility in plants decreased in the order Zn>Cd>Pb; it was generally lower in heavily polluted areas. The content of all the three metals in plant roots correlated with topsoil concentration of their 2 mol/L HNO₃-extractable forms. Besides, Cd and Zn correlated with exchangeable and Zn also with the organic form content. The concentration factor as a measure of metal mobility from soil to plants decreased with increasing level of contamination. Presented at the 4th Mini-Symposium on Biosorption and Microbial Degradation, Prague, Czech Republic, November 26–29, 1996.     

The proline biosynthesis in living organisms

Summary In this article we review recent work on the physiology of proline and ¹-pyrroline-5-carboxylate (P5C) in living organisms and consider recent progress in our understanding of the role of P5C synthetase in collagen metabolism and the regulation of urea cycle in vertebrates. Much of this recent progress has been made possible by advances in our knowledge of the enzymes and genes involved in proline biosynthesis in man. The availability of well characterized P5C synthetase deficiency in man has been an impetus for the cloning of the cDNA encoding for this enzyme from man and facilitated the establishment of the phenotype-genotype relationships in P5C synthetase deficiency in higher vertebrates.Abbreviations GK -glutamyl kinase - GPR -glutamyl phosphate reductase - P5CR ¹-pyrroline-5-carboxylate reductase - GSA glutamic--semialdehyde - P5C ¹-pyrroline-5-carboxylate - P₁ Inorganic phosphate - AMP, ADP, ATP Adenosine 5-mono-, di-, triphosphate - NAD⁺, NADH nicotinamide adenine dinucleotide, and its reduced form - NADP⁺, NADPH nicotinamide adenine dinucleotide phosphate, and its reduced form; DEAF: diethylaminoethyle - OAT ornithine amino transferase; CHO: Chinese hamster ovary - IGF-1 insulin-like growth factor-1 - P5CDH pyrroline 5carboxylate dehydrogenase - IMP inosine 5-monophosphate     

Unravelling the mechanisms for plant survival on gypsum soils: an analysis of the chemical composition of gypsum plants from Turkey

Depending on their specificity to gypsum, plants can be classified as gypsophiles (gypsum exclusive) and gypsovags (non‐exclusive). The former may further be segregated into wide and narrow gypsophiles, depending on the breadth of their distribution area. Narrow gypsum endemics have a putative similar chemical composition to plants non‐exclusive to gypsum (i.e. gypsovags), which may indicate their similar ecological strategy as stress‐tolerant plant refugees on gypsum. However, this hypothesis awaits testing in different regions of the world. We compared the chemical composition of four narrow gypsum endemics, one widely distributed gypsophile and six gypsovags from Turkey. Further, we explored the plasticity in chemical composition of Turkish gypsovags growing on high‐ and low‐gypsum content soils. Differences were explored with multivariate analyses (RDA) and mixed models (REML). Narrow gypsum endemics segregated from gypsovags in their chemical composition according to RDAs (mainly due to higher K and ash content in the former). Nevertheless, differences were small and disappeared when different nutrients were analysed individually. All the gypsovags studied accumulated more S and ash when growing on high‐gypsum than on low‐gypsum soils. Similar to narrow gypsum endemics from other regions of the world, most local gypsum endemics from Turkey show a similar chemical composition to gypsovags. This may indicate a shared ecological strategy as stress‐tolerant plants not specifically adapted to gypsum. Nevertheless, the narrow gypsum endemic Gypsophila parva showed a chemical composition typical of gypsum specialists, indicating that various strategies are feasible within narrowly distributed gypsophiles.     

Plants living on gypsum: beyond the specialist model

BACKGROUND AND AIMS: Plants from gypsum habitats are classified as gypsophiles and gypsovags. The former include both narrow endemics limited to small gypsum areas and regionally dominant gypsophiles growing in gypsum areas of large regions, whereas gypsovags are plants that can grow both in gypsum and non-gypsum soils. Factors controlling the distribution of gypsum plants are still not fully understood. METHODS: To assess how the different types of gypsum plants deal with the stressful conditions of gypsum substrates, comparisons were made of the leaf chemical composition of four gypsovags, five regionally dominant gypsophiles and four narrow gypsum endemics growing in two massive gypsum areas of the Iberian Peninsula. KEY RESULTS: The chemical composition of gypsovags was clearly different from regionally dominant gypsophiles, while the chemical composition of narrow-gypsophile endemics was more similar to the chemical composition of gypsovags than to that of regionally dominant gypsophiles. Regionally dominant gypsophiles showed higher concentrations of ash, Ca, S, N, Mg P and Na, whereas gypsovags and local gypsophile endemics displayed higher concentrations of C and greater C : N ratios. CONCLUSIONS: Such differences suggest that the three groups of gypsum plants follow diverse ecological strategies. It is suggested that regionally dominant gypsophiles might fit the 'specialist' model, being species specifically adapted to gypsum, whereas both gypsovags and narrow-gypsophile endemics might fit the 'refuge' model, being stress-tolerant species that find refuge on gypsum soils from competition. The analysis of the leaf chemical composition could be a good predictor of the degree of plants specialization to gypsum soils.     

The effect of NaCl on proline accumulation in rice leaves

The regulation of proline accumulation in detached leaves of rice(Oryza sativa cv. Taichung Native 1) was investigated.Increasing concentrations of NaCl from 50 to 200 mM progressivelyincreased proline content in detached rice leaves. NaCl induced prolineaccumulation was mainly due to the effect of both Na⁺ andCl^– ions. Proline accumulation caused by NaCl was related toprotein proteolysis, an increase in ornithine--aminotransferaseactivity,a decrease in proline dehydrogenase activity, a decrease in prolineutilisation,and an increase in the content of the precursors of proline biosynthesis,ornithine and arginine. Results also show that proline accumulation caused byNaCl was associated with ammonium ion accumulation.     

Thioproline-induced accumulation of proline in barley leaf sections

Abstract Treatment of barley leaf sections with 0.1 mol m^?3 thioproline (L-thiazolidine-4-carboxylic acid) was found to induce a marked increase in proline together with some decrease in glutamate, whereas the levels of other andno acids were not influenced. This result is discussed in relation to the significance of the increase in proline in tissues treated with abscisic acid or subjected to water stress.     

Abscisic Acid accumulation is not required for proline accumulation in wilted leaves

Leaves from dark-grown barley (Hordeum vulgare L. var Larker) seedlings grown in the presence and absence of fluridone were used to determine whether or not abscisic acid (ABA) accumulation was necessary for proline to accumulate in wilted tissue. Wilted tissue (polyethylene glycol-treated) leaves from fluridone-grown seedlings did not accumulate ABA but did accumulate proline at a rate that was not different from the non-fluridone-treated leaves. Thus ABA accumulation is not required for wilting-induced proline accumulation in barley leaves. Proline accumulation in wilted leaves from the wilty tomato (Lycopersicon esculentum) mutant, flacca, was compared to that in the wild type, Rheinlands Ruhm. Proline accumulated in wilted leaves from flacca. The rate of accumulation was faster in flacca compared to the rate in the wild type because the wilty mutant wilted faster. ABA accumulated in wilted leaves from the wild type but not in the wilty mutant. This result is a further confirmation that ABA accumulation is not required for wilting-induced proline accumulation. These results are significant in that proline accumulation in barley leaves can be induced independently by any one of three treatments: wilting, ABA, or salt.     

Toxicity and accumulation of copper and nickel in maize plants cropped on calcareous and acidic field soils

Field experiments in calcareous and acidic field soils were conducted to study the effects of copper (Cu) and nickel (Ni) added to soils on maize growth and metal accumulation in maize plants. The results revealed that the critical concentrations of Cu added to soils that decreased maize grain yield by 10% (EC₁₀) were 711 mg kg^?1 for calcareous soil with a pH of 8.9, and 23 mg kg^?1 for acidic soil with a pH of 5.3. The toxicity thresholds of EC₁₀ did not differ significantly for Cu and Ni. A different pattern of Cu and Ni accumulation in maize plants was also found. The accumulation of Cu in above-ground parts of the plants increased initially as the concentrations of Cu added to soils increased, after which they decreased to a constant level. As the concentrations of Ni added to soils increased, the accumulation of Ni in stems and leaves increased linearly, but the accumulation of Ni in the grains was nonlinear. Additionally, the results revealed that Ni was transported to grains more easily than Cu. The results also showed that the concentrations of Cu and Ni in soil solutions as toxicity predictors and the critical concentrations of Cu and Ni in maize were all soil-dependent.     

Importance of N source on heat stress tolerance due to the accumulation of proline and quaternary ammonium compounds in tomato plants

Proline and quaternary ammonium compounds (QAC), in addition to being N-rich, are known to accumulate in plants under different environmental stress conditions. The accumulation of N-rich compounds in plants has been shown to confer stress resistance. The aim of our work is two-fold: first, to study the influence of temperature on proline, QAC, and choline metabolism in tomato leaves; and second, to investigate the relationship between N source applied (NO3- or NH4+) and thermal stress resistance in these plants. To do this, experiments were conducted at three different temperatures (10 degrees C, 25 degrees C, 35 degrees C); at each temperature half of the plants received NO3-, and the other half received NH4+. At 35 degrees C the plants had the lowest biomass production with respect to 25 degrees C (optimal temperature) and 10 degrees C (cold stress), suggesting that tomato plants were most affected by heat stress. At 35 degrees C, there were also high levels of choline and proline due to the activation of Delta1-pyrroline-5-carboxylate synthetase (P5CS) and ornithine aminotransferase (OAT), and simultaneous inhibition of proline dehydrogenase (PDH) and proline oxidase (PO). However, plants with NH4+ as the N source exhibited reduced growth with respect to the plants fed with NO3-. This is interesting because, under heat stress (35 degrees C), biomass production, as well as proline and choline accumulation, in NH4+ fed plants was higher than in NO3- fed plants. From this, we concluded that tomato plants fed with NH4+ as the N source show higher tolerance to heat stress (35 degrees C) than plants fed with NO3-.     

Diurnal changes in proline content of desert plants

Summary The diurnal changes in proline and water contents of Zygophyllum quatarense and Francoeuria crispa were investigated under desert conditions. In both plants, the proline content was low before sunrise amounting to 10.42 moles/g fresh weight in Zygophyllum and 6.3 moles/g fresh weight in Francoeuria. By the progress of the day and the intensification of the evaporative power of the atmosphere, the proline content increased considerably in the two plants. The proline content reached maximal values at 5 p.m., reaching 23.36 moles/g in Zygophyllum and 11.16 moles/g in Francoeuria. The proline content of the artificially watered plants was kept at a low level throughout the day. The conditions and role of proline accumulation were discussed.     

Regulation of ammonium-induced proline accumulation in detached rice leaves

Accumulation of proline in response to NH₄Cl was studied indetached leaves of rice (Oryza sativa cv. Taichung Native1). Increasing concentrations of NH₄Cl from 50 to 200mMprogressively increased proline content and this was correlated with theincrease in ammonium content. Proline accumulation induced by NH₄Clwas related to proteolysis, an increase in ornithine--aminotransferaseactivity, a decrease in proline dehydrogenase activity, and a decrease inproline utilisation and could not be explained by NH₄Cl-inducedmodification in ¹-pyrroline-5-carboxylate reductase activity.The content of glutamic acid was decreased by NH₄Cl, whereas theincrease in arginine and ornithine contents was found to be associated with theincrease in proline content in NH₄Cl-treated detached rice leaves.     

Role of carbohydrates in proline accumulation in wilted barley leaves

The effect of wilting on proline synthesis, proline oxidation, and protein synthesis—all of which contribute to proline accumulation—was determined in nonstarved barley (Hordeum vulgare L.) leaves. Nonstarved leaves were from plants previously in the light for 24 hours and starved leaves were from plants previously in the dark for 48 hours. Wilted leaves from nonstarved plants accumulated proline at the rate of about 1 μmole per hour per gram of fresh weight whereas wilted leaves from starved plants accumulated very little proline. Wilting caused a 40-fold stimulation of proline synthesis from glutamate in nonstarved leaves but had very little effect in starved leaves. Proline oxidation and protein synthesis, on the other hand, were inhibited by wilting in both nonstarved and starved leaves. Thus, the role of carbohydrates in proline accumulation is to supply precursors for the stimulated proline synthesis. These results further indicate that the main metabolic response causing proline to accumulate in wilted barley leaves is the stimulation of proline synthesis from glutamate. The difference between these results and those obtained with beans is discussed.

Wilting caused an increased conversion of glutamate to other products. In nonstarved leaves, conversion to organic acids as well as to proline was increased. In starved leaves, wilting caused an increase in the conversion of glutamate to glutamine, aspartate, asparagine, and organic acids.