Solutes Glycine Betaine Research Articles

Does habitat of Atriplex halimus L. affect plant strategy for osmotic adjustment?

The impact of water stress was analysed in the xero-halophyte Mediterranean shrub Atriplex halimus using two Tunisian populations originating from a sub-humid coastal site (Monastir) or from a semi-arid area (Kairouan). Seedlings were exposed for 10 days to nutrient solution containing either 0 or 15% polyethylene glycol. Water potential (Ψw), osmotic potential (Ψs), osmotic potential at full turgor [Ψs(100)], relative water content (RWC), shoot dry weight (DW) and changes in solute concentrations were quantified every 2 days throughout the stress period and inorganic solutes contents were determined at the end of the treatment. The water deficit induced a decrease in Ψw, Ψs and RWC in both populations, recorded changes being higher in plants of Monastir than those of Kairouan while the shoot dry weight was reduced in a similar extent in stressed plants from both populations. Water deficit induced an increase in proline, glycinebetaine and sugar concentrations. Proline accumulated as early as after the 24-h stress treatment while, glycinebetaine required more than 6 days of stress to accumulate. At the end of the stress period, the plants of Kairouan population accumulated higher amounts of proline than those of Monastir, while an opposite trend was reported for glycinebetaine. Both populations specifically accumulated Na+ in response to drought stress, suggesting that this element could play a physiological role in the stress response of this xero-halophyte species. Presented results suggest that the non-recyclable osmotic solute glycinebetaine does not necessarily preferentially accumulates in population facing permanent water stress and that other strategy than osmotic adjustment might be involved in drought tolerance of A. halimus.

A comparative study of the efficiency of several sources of induced resistance to powdery mildew ( Blumeria graminis f. sp. tritici) in wheat under field conditions

In comparison with untreated controls, both synthetic and biologically sourced inducers of resistance significantly decreased the severity of powdery mildew in a 3-year, small-plot experiment which included other cultivars with varying resistance to powdery mildew as reference, on the susceptible winter wheat cultivar, Kanzler. The most prominent effects were observed following treatments with benzothiadiazole, extracts made of ginger ( Zingiber officinale Roscoe) and curcuma ( Curcuma longa L.) rhizomes, and giant knotweed ( Reynoutria sacchaliensis L.) leaves, which were highly efficient in all 3 years of the experiment. Other inducers such as oak bark ( Quercus robur L.) extract, salicylic acid as well as a solution of glycine betaine were also effective. All inducers under study had halted disease progress by 27 d after application. The induced resistance was long-lasting and the severity of the disease, when compared with the untreated control, was decreased to between 2% and 53%, depending upon the year.

Physiological Role of Exogenously Applied Glycinebetaine to Improve Drought Tolerance in Fine Grain Aromatic Rice (Oryza sativa L.)

AbstractRice performance under drought stress is mainly impeded by oxidative damage and hampered plant water status, which may be improved by exogenous use of osmoprotectants. In this study, the role of glycinebetaine (GB) to improve drought tolerance in rice (Oryza sativa L.) cultivar Super‐basmati was evaluated. GB was used both as seed and foliar application. For priming, seeds were soaked in 50, 100 and 150 mg l−1 aerated solution of GB for 48 h. At four‐leaf stage, one set of plants was subjected to drought stress, while the other set kept at full field capacity. Drought was maintained at 50 % of field capacity by watering when needed. For exogenous application, 50, 100 and 150 mg l−1 GB levels were applied at five‐leaf stage. Drought stress greatly reduced the rice growth while GB application improved it both under well‐watered and drought conditions. Drought tolerance in rice was strongly related to the maintenance of tissue water potential and antioxidant system, which improved the integrity of cellular membranes and enabled the plant to maintain high photosynthesis. Foliar treatments were more effective than the seed treatments, while among the GB treatment, foliar application with 100 mg l−1 was the most effective.

Glycinebetaine Improves Chilling Tolerance in Hybrid Maize

AbstractPlant growth and development is hampered by various environmental stresses including chilling. We investigated the possibility of improving chilling tolerance in hybrid maize by glycinebetaine (GB) seed treatments. Maize hybrid (Hycorn 8288) seeds were soaked in 50, 100 and 150 mg l−1(p.p.m.) aerated solution of GB for 24 h and were dried back. Treated and untreated seeds were sown at 27 °C (optimal temperature) and at 15 °C (chilling stress) under controlled conditions. Germination and seedling growth was significantly hindered under chilling stress. Moreover, chilling stress significantly reduced the starch metabolism and relative water contents (RWC), and increased the membrane electrolyte leakage. However, activities of antioxidants (catalase, superoxide dismutase and ascorbate peroxidase) were increased under stress conditions. Seed treatments with GB improved the germination rate, root and shoot length, seedling fresh and dry weights, leaf and root scores, RWC, soluble sugars, α‐amylase activity and antioxidants significantly compared with untreated seeds under optimal and stress conditions. Induction of chilling tolerance was attributed to maintenance of high tissue water contents, reduced membrane electrolyte leakage, and higher antioxidant activities and carbohydrate metabolism. Seed treatment with 100 mg l−1GB was the best treatment for improving the performance of hybrid maize under normal and stress conditions compared with control and other levels used.

Comparative Study of Urea and Betaine Solutions by Dielectric Spectroscopy: Liquid Structures of a Protein Denaturant and Stabilizer

We performed dielectric spectroscopy measurements on aqueous solutions of glycine betaine (N,N,N-trimethylglycine), which is known to be a strong stabilizer of globular proteins, over a wide concentration range (3-62 wt %) and compared the results with our previously published data for aqueous solutions of urea, a representative protein denaturant. The hydration number of betaine (9), calculated on the basis of the reduction in the dielectric relaxation strength of bulk water with addition of betaine, is significantly larger than that of urea (2). Furthermore, the dielectric relaxation time increased with betaine concentration, while that remained nearly constant for the urea-water system over a wide concentration range. This difference between urea and betaine is probably related to their opposite effects on the protein stabilization.

High concentrations of the compatible solute glycinebetaine destabilize model membranes under stress conditions

Compatible solutes are accumulated by diverse organisms in response to environmental stresses such as drought, salt, or cold. Glycinebetaine (Bet) is such a solute that is accumulated by many plants and microorganisms to high concentrations under stress conditions. It is an osmoprotectant in bacteria and stabilizes both soluble and peripherally membrane-bound proteins in vitro. Here, the effects of Bet on the stability of model lipid membranes are compared to the effects of two other compatible solutes, sucrose and trehalose. Both in the presence of 1 M NaCl and during freezing to −20 °C, Bet is highly destabilizing to liposomes containing nonbilayer lipids, while the disaccharides are either protective or, in some cases, much less destabilizing. The destabilizing effect of Bet is more pronounced in membranes containing the nonbilayer galactolipid monogalactosyldiacylglycerol from plant chloroplasts than in membranes containing the nonbilayer phospholipid phosphatidylethanolamine. The most dramatic differences between the sugars and Bet were observed in liposomes made from a combination of lipids resembling plant chloroplast thylakoid membranes. Measurements with the dye merocyanine 540 indicate that the water-membrane interface was affected in opposite directions by the presence of high concentrations of sucrose or Bet. The dynamics of the lipids, however, were not differentially affected by the solutes, making direct solute-lipid interactions an unlikely explanation for the different effects on stability. The data offer an explanation, why Bet at high concentrations achieved during exogenous feeding of leaf tissues can be detrimental to cellular stability and survival under stress, while bacterial membranes that contain phosphatidylethanolamine instead of monogalactosyldiacylglycerol, or cyanobacteria that contain highly saturated monogalactosyldiacylglycerol are less susceptible.

Accumulation of the compatible solutes, glycine-betaine and ectoine, in osmotic stress adaptation and heat shock cross-protection in the biocontrol agent Pantoea agglomerans CPA-2

The effect of modifying the water activity (a(w)) of Pantoea agglomerans growth medium with the ionic solute NaCl on water stress resistance, heat-shock survival and intracellular accumulation of the compatible solutes glycine-betaine and ectoine were determined. The bacterium was cultured in an unmodified liquid medium or that modified with NaCl to 0.98 and 0.97 a(w), and viability of cells evaluated on a 0.96 a(w)-modified solid media to check water stress tolerance. Cells grown under ionic stress had better water stress tolerance than control cells. These cells also had cross-protection to heat stress (30 min, 45 degrees C). The modified cells accumulated substantial amounts of the compatible solutes glycine-betaine and ectoine in contrast to the control cells, which contained little or none of these two compounds. Improvement in osmotic and thermal tolerance of cells of the biocontrol agent P. agglomerans by modifying growth media with the ionic solute NaCl was achieved. The compatible solutes glycine-betaine and ectoine play a critical role in environmental stress tolerance improvement. This approach provides a method for improving the physiological quality of inocula and could have implications for formulation and shelf-life of biocontrol agents.

Dielectric Behavior of Aqueous Micellar Solutions of Betaine-Type Surfactants

Dielectric behavior was examined for aqueous solutions of the betaine-type surfactants dodecyldimethylcarbobetaine (C(12)DCB), tetradecyldimethylcarbobetaine (C(14)DCB), cetyldimethylcarbobetaine (C(16)DCB), and oleyldimethylcarbobetaine (OleyDCB) as a function of frequency from 1.00 x 10(6) to 2.00 x 10(10) Hz (6.28 x 10(6) to 1.26 x 10(11) rad s(-1)) with changing surfactant concentration (c(D)). Rotational relaxation times (tau) of the zwitterionic headgroups of the surfactants in aqueous solutions of C(12)DCB and C(14)DCB, which form spherical micelles, are determined to be 0.26 and 0.30 ns, respectively. Values of tau for aqueous solutions of C(16)DCB and OleyDCB, which form threadlike micelles, are identical at 0.44 ns. The tau values of all micellar solutions are constant irrespective of c(D). The increase in tau with increasing alkyl chain length is assigned to an increase of molecular density at the micellar surface. The magnitude of the relaxation strength for the surfactant solutions increases in proportion to c(D) and is not so different from that of an aqueous solution of glycine betaine (GB), which has the same chemical structure as betaine-type surfactants with zwitterionic headgroups but never forms micelles. This finding suggests that the zwitterionic headgroup rotating on the micellar surface possesses a dipole moment with a magnitude essentially the same as that of GB in aqueous solutions.

Role of the glycine betaine and carnitine transporters in adaptation of Listeria monocytogenes to chill stress in defined medium.

The food-borne pathogen Listeria monocytogenes proliferates at refrigeration temperatures, rendering refrigeration ineffective in the preservation of Listeria-contaminated foods. The uptake and intracellular accumulation of the potent compatible solutes glycine betaine and carnitine has been shown to be a key mediator of the pathogen's cold-tolerant phenotype. To date, three compatible solute systems are known to operate in L. monocytogenes: glycine betaine porter I (BetL), glycine betaine porter II (Gbu), and the carnitine transporter OpuC. We investigated the specificity of each transporter towards each compatible solute at 4 degrees C by examining mutant derivatives of L. monocytogenes 10403S that possess each of the transporters in isolation. Kinetic and steady-state compatible solute accumulation data together with growth rate experiments demonstrated that under cold stress glycine betaine transport is primarily mediated by Gbu and that Gbu-mediated betaine uptake results in significant growth stimulation of chill-stressed cells. BetL and OpuC can serve as minor porters for the uptake of betaine, and their action is capable of providing a small degree of cryotolerance. Under cold stress, carnitine transport occurs primarily through OpuC and results in a high level of cryoprotection. Weak carnitine transport occurs via Gbu and BetL, conferring correspondingly weak cryoprotection. No other transporter in L. monocytogenes 10403S appears to be involved in transport of either compatible solute at 4 degrees C, since a triple mutant strain yielded neither transport nor accumulation of glycine betaine or carnitine and could not be rescued by either osmolyte when grown at that temperature.

Molecular dynamics simulation of aqueous solutions of glycine betaine

Molecular dynamics simulation is used to investigate hydration properties of glycine betaine in a large range of solute concentrations. Statistical analyses of the system trajectories evidence microscopic details suggesting an interpretation of experimental results recently obtained for aqueous solutions of trimethylamine- N-oxide, a bioprotectant closely related to glycine betaine.

Salinity Tolerance Mechanisms of Grasses in the Subfamily Chloridoideae

Forage grasses and turfgrasses are increasingly being subjected to salinity stress, due to accelerated salinization of irrigated agricultural lands worldwide, and to increased use of reclaimed and other secondary water sources for irrigating turfgrass landscapes. The objective of this study was to examine salinity responses of a number of important forage and turfgrass genera in the subfamily Chloridoideae in attempt to gain understanding of salinity tolerance mechanisms operating in this subfamily. Grasses were exposed to salinities up to 600 mM NaCI in solution culture. Salinity tolerance decreased in the following order: Distichlis spicata vat. stricta (Tort.) Beetle > Sporobolus airoides (Tort.) Tort. > Cynodon dactylon (L.) Pets. = Zoysia japonica Steud. > Sporobolus cryptandrus (Tort.) A. Gray. Buchloë dactyloides (Nutt.) Engelm. > Bouteloua curtipendula (Michx.) Tort. Relative root length (RL) and relative root weight (RW) increased under saline conditions, relative to control, in salt tolerant grasses. Leaf sap osmolality, Na+, Cl−, and proline concentrations were negatively correlated and glycinebetaine was positively correlated with salinity tolerance. Bicellular salt glands were observed on leaves of all species. Salinity tolerance was positively correlated with Na+ and Cl− salt gland secretion rates. Within the subfamily Chloridoideae, salinity tolerance was associated with saline ion exclusion, facilitated by leaf salt gland ion secretion, and with accumulation of the compatible solute glycinebetaine.

The Storage Life of Packed Shredded Iceberg Lettuce Dipped in Glycine Betaine Solutions

The effects of glycine betaine dip, packaging method, and storage time on the sensory quality of shredded Iceberg lettuce were first modeled using a statistical experimental design, followed by a second storage test verifying the effect of the glycine betaine treatment. Shredded lettuce was dipped in 0 to 100 mg/liter active chlorine solution and then in 0 to 1.0 mol/liter glycine betaine solution, packed in 25μm oriented polypropylene film, 250 g per package, and stored at 5°C for 8 days. Models with good predictability were created suggesting that the glycine betaine dip helped retain sensory quality, especially appearance (P < 0.05). The models also suggested that washing periods over 60 s were not needed and that the microperforation of packages should not exceed 0.31 mm2 per package. The modeled positive effect on sensory quality was verified in the second storage test (P < 0.05). The optimum glycine betaine concentration was 0.2 mol/liter. Chlorination of the first dip particularly retained appearance of packed lettuce.

Isolation of the putP gene of Corynebacterium glutamicum and characterization of a low-affinity uptake system for compatible solutes

Corynebacterium glutamicum accumulates the compatible solutes proline, glycine betaine, and ectoine under conditions of high osmolality. Uptake of proline is mediated by both a high-affinity and a low-affinity secondary transport system. The low-affinity uptake system also accepts glycine betaine and ectoine as substrates. In the present study, the gene encoding the high-affinity proline uptake system PutP was isolated by heterologous complementation of Escherichia coli mutant strain WG389, which lacks the transport systems BetT, PutP, ProP, and ProU and is unable to synthesize proline and glycine betaine. This gene (putP) encodes a protein of 524 amino acids that shares identity with the proline transport systems PutP of E. coli, Staphylococcus aureus, Salmonella typhimurium, Haemophilus influenzae, and Klebsiella pneumoniae. Functional studies of PutP synthesized in E. coli mutant strain MKH13, which also lacks the transport systems for compatible solutes and is unable to synthesize glycine betaine, revealed that this carrier system is not regulated by the external osmolality on the level of activity. Km values of 7.6 mM for proline and 1.3 mM for sodium as cotransported ion were determined. Deletion of the putP gene allowed the functional characterization of another proline uptake system with low affinity.

Uptake and translocation of foliar-applied glycinebetaine in crop plants

Glycinebetaine is known as an osmoprotectant which is accumulated in certain plant species under salt and drought stresses. Exogenous applications of glycinebetaine on crop plants unable to synthesise glycinebetaine is a possible approach to overcome the environmental limitations of crop production. Information about the capability of plants to take up and translocate foliar-applied glycinebetaine is, however, limited. In this study, glycinebetaine solution, with and without surfactants, was exogenously applied to foliage of summer turnip rape ( Brassica rapa L. ssp. oleifera), soybean [ Glycine max (L.) Merr.], pea ( Pisum sativum L.), tomato ( Lycopersicon esculentum Mill.), and spring wheat ( Triticum aestivum L.) in greenhouse experiments. Uptake of glycinebetaine was monitored using high-performance liquid chromatography. [ 14C]glycinebetaine was applied to leaves of turnip rape plants and translocation to other plant organs was monitored autoradiographically. [ 14C]glycinebetaine was translocated to roots within two hours of application. One day after application labelled glycinebetaine was translocated to all plant parts of turnip rape plants. The results indicate that plants are able to translocate foliar-applied glycinebetaine from their leaves to other organs, and that the use of surfactants accelerates the penetration of foliar-applied glycinebetaine. According to our results glycinebetaine is quite inert end-product in plant cells being mainly phloem-mobile. Moreover, environmental conditions are shown to affect the uptake and translocation rates of foliar-applied glycinebetaine.

Biosynthesis and fate of compatible solutes in extremely halophilic phototrophic eubacteria

Most halophilic and halotolerant eubacteria are able to accumulate compatible solutes from their environment during salt stress. They are usually able to synthesize sugars and amino acids. These compatible solutes, however, are of less importance in extremely halophilic eubacteria, where usually glycinebetaine or ectoine is required. Extremely halophilic phototrophic sulfur bacteria of the genus Ectothiorhodospira are able to synthesize the three compatible solutes glycinebetaine, trehalose and ectoine. While glycinebetaine is the major compatible solute under all conditions the percentage of trehalose and ectoine varies depending on the availability of nitrogen sources. Using acetate plus bicarbonate as simultaneous substrates the three compatible solutes were analysed by 13C-NMR spectroscopy. The label found indicated the following biosynthesis pathways: glycine, derived from glyoxylate out of the Kornberg cycle, undergoes a three-fold methylation with S-adenosylmethionine as methyl donor. The latter is derived from the tetrahydrofolate pathway. Several enzymes of this pathway have been found and are under investigation. The labelling of trehalose indicates that the Calvin cycle is blocked in the presence of acetate. The two glucose moieties of trehalose are linked by trehalose-6-phosphate synthase. The enzyme was characterized. Ectoine is synthesized from aspartate via aspartophosphate, aspartate semialdehyde and α, γ-diaminobutyrate. Dilution stress leads to rapid excretion of betaine and ectoine, followed by immediate uptake to balance overshoot excretion. Trehalose is not excreted under dilution stress but is degraded by trehalase and subsequently metabolized.

Biosynthesis and fate of compatible solutes in extremely halophilic phototrophic eubacteria

Most halophilic and halotolerant eubacteria are able to accumulate compatible solutes from their environment during salt stress. They are usually able to synthesize sugars and amino acids. These compatible solutes, however, are of less importance in extremely halophilic eubacteria, where usually glycinebetaine or ectoine is required. Extremely halophilic phototrophic sulfur bacteria of the genus Ectothiorhodospira are able to synthesize the three compatible solutes glycinebetaine, trehalose and ectoine. While glycinebetaine is the major compatible solute under all conditions the percentage of trehalose and ectoine varies depending on the availability of nitrogen sources. Using acetate plus bicarbonate as simultaneous substrates the three compatible solutes were analysed by 13C-NMR spectroscopy. The label found indicated the following biosynthesis pathways: glycine, derived from glyoxylate out of the Kornberg cycle, undergoes a three-fold methylation with S-adenosylmethionine as methyl donor. The latter is derived from the tetrahydrofolate pathway. Several enzymes of this pathway have been found and are under investigation. The labelling of trehalose indicates that the Calvin cycle is blocked in the presence of acetate. The two glucose moieties of trehalose are linked by trehalose-6-phosphate synthase. The enzyme was characterized. Ectoine is synthesized from aspartate via aspartophosphate, aspartate semialdehyde and α, β-diaminobutyrate. Dilution stress leads to rapid excretion of betaine and ectoine, followed by immediate uptake to balance overshoot excretion. Trehalose is not excreted under dilution stress but is degraded by trehalase and subsequently metabolized.

The effect of salinity on the yield and composition of Diplachne fusca (Kallar grass)

Abstract. Aspects of the response of Kallar grass (Diplachne fusca) to root salinity have been studied. Field observation on the high level of tolerance of this species have been mirrored in pot trials and a 50% yield (dry matter) was obtained at ECe 22.3. While salt stress led to Na and Cl uptake, most of these ions appeared to be secreted selectively from the leaves. The shoot K+ content on a tissue water basis remained unaffected by salt stress and the shoot tissue had a high K+ selectivity. Osmotic adaptation was mainly brought about by tissue dehydration and not net salt accumulation although the compatible solute glycinebetaine was accumulated in fairly high concentrations.The organic and inorganic chemical composition of the shoots, including the trace elements Zn, Fe, Cu and Mn, is such as to make this species a highly promising plant for the exploitation of saline soils which might otherwise not be amenable to agriculture use.