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Glycine betaine is an osmoprotectant compound which enhances cell tolerance in plant species in response to environmental stresses. This study aimed to investigate the effect of exogenous application of glycine betaine on some antioxidant activities of tobacco plants overexpressing P5CS gene. Sterile tobacco seedlings with four to six leaves were transferred to MS medium containing 0, 100, and 200 mM NaCl, after which glycine betaine (20 and 40 mg l-1) were foliar sprayed on the surface of the plants. After four weeks, glycine betaine treatment enhanced the antioxidant capacity of the plant through activation of catalase (CAT), superoxide dismutase (SOD), and ascorbate peroxidase (APX). In contrast, H2O2 content and MDA level were reduced by glycine betaine under similar conditions. Therefore, application of exogenous glycine betaine under salt stress improved stress tolerance in T and non-T plants. Meanwhile, our results indicated the positive effect of glycine betaine in T plants was greater than in non-T plants. On the other hand, this result suggested that the synergistic effects of glycine betaine and proline in plants enhanced the antioxidant defense system in T plants overexpressing P5CS gene.

Soil salinity is a major constraint to wheat production; it causes a severe reduction in wheat growth and yield. Alleviation of salinity effects on physiological, biochemical, and yield of wheat cultivars; Sids 14 and Misr 3 using some soil additions (control, Molasses and Humic acid), compatible solutes, and growth regulators (water as control, Naphthalene acetic acid, and Glycine betaine) were investigated in salt-affected soils. Results indicated that Misr 3 was superior to Sids 14 in all studied characteristics except flag leaf area, relative water content, plant height and recorded lower and desirable value of leaf temperature. The addition of Molasses (24 L ha−1) or Humic acid (12 L ha−1) significantly increased physiological and biochemical characteristics. At the same time, flag leaf temperature, proline, and malondialdehyde (MDA) content were decreased, yield and its attributes also increased except No. kernel spike-1. Foliar spray of Naphthalene acetic acid (NAA) at 30 mg L−1. or glycine betaine (GB 100 mM) also positively affected the studied characteristics, where Glycine betaine recorded the highest Relative water content and Fv/Fm. In contrast, NAA recorded the most increased Catalase (CAT) activity, and the Number of spikes m−2 and insignificant differences were observed between them in grain yield. It could be recommended the cultivation of Misr 3 with Molasses and GB under saline soils.

Bacteria have evolved mechanisms that allow them to adapt to changes in osmolarity and some species have adapted to live optimally in high salinity environments such as in the marine ecosystem. Most bacteria that live in high salinity do so by the biosynthesis and/or uptake of compatible solutes, small organic molecules that maintain the turgor pressure of the cell. Osmotic stress response mechanisms and their regulation among marine heterotrophic bacteria are poorly understood. In this review, we discuss what is known about compatible solute metabolism and transport and new insights gained from studying marine bacteria belonging to the family Vibrionaceae.

Background Application of exogenous glycine betaine (GB) and exogenous salicylic acid (SA) mitigates the adverse effects of salinity. Foliar spraying with exogenous GB or SA alleviates salt stress in plants by increasing leaf gas exchange and stimulating antioxidant enzyme activity. The effects of foliar application of exogenous GB and SA on the physiology and biochemistry of cotton seedlings subjected to salt stress remain unclear. Results Results showed that salt stress of 150 mM NaCl significantly reduced leaf gas exchange and chlorophyll fluorescence and decreased photosynthetic pigment quantities and leaf relative water content. Foliar spray concentrations of 5.0 mM exogenous GB and 1.0 mM exogenous SA promoted gas exchange and fluorescence in cotton seedlings, increased quantities of chlorophyll pigments, and stimulated the antioxidant enzyme activity. The foliar spray also increased leaf relative water content and endogenous GB and SA content in comparison with the salt-stressed only control. Despite the salt-induced increase in antioxidant enzyme content, exogenous GB and SA in experimental concentrations significantly increased the activity of glutathione reductase, ascorbate peroxidase, superoxide dismutase, catalase and peroxidase, and decreased malondialdehyde content under salt stress. Across all experimental foliar spray GB and SA concentrations, the photochemical efficiency of photosystem II (FV/FM) reached a peak at a concentration of 5.0 mM GB. The net photosynthetic rate (Pn) and FV/FM were positively correlated with chlorophyll a and chlorophyll b content in response to foliar spraying of exogenous GB and SA under salt stress. Conclusions We concluded, from our results, that concentrations of 5.0 mM GB or 1.0 mM SA are optimal choices for mitigating NaCl-induced damage in cotton seedlings because they promote leaf photosynthesis, increase quantities of photosynthetic pigments, and stimulate antioxidant enzyme activity. Among, 5.0 mM GB and 1.0 mM SA, the best performance in enhancing endogenous GB and SA concentrations was obtained with the foliar application of 1.0 mM SA under salt stress.

Among compatible solutes, glycine betaine has various applications in the fields of nutrition, pharmaceuticals, and cosmetics. Currently, this compound can be extracted from sugar beet plants or obtained by chemical synthesis, resulting in low yields or high carbon footprint, respectively. Hence, in this work we aimed at exploring the production of glycine betaine using the unicellular cyanobacterium Synechocystis sp. PCC 6803 as a photoautotrophic chassis. Synechocystis mutants lacking the native compatible solutes sucrose or/and glucosylglycerol—∆sps, ∆ggpS, and ∆sps∆ggpS—were generated and characterized. Under salt stress conditions, the growth was impaired and accumulation of glycogen decreased by ∼50% whereas the production of compatible solutes and extracellular polymeric substances (capsular and released ones) increased with salinity. These mutants were used as chassis for the implementation of a synthetic device based on the metabolic pathway described for the halophilic cyanobacterium Aphanothece halophytica for the production of the compatible solute glycine betaine. Transcription of ORFs comprising the device was shown to be stable and insulated from Synechocystis’ native regulatory network. Production of glycine betaine was achieved in all chassis tested, and was shown to increase with salinity. The introduction of the glycine betaine synthetic device into the ∆ggpS background improved its growth and enabled survival under 5% NaCl, which was not observed in the absence of the device. The maximum glycine betaine production [64.29 µmol/gDW (1.89 µmol/mg protein)] was reached in the ∆ggpS chassis grown under 3% NaCl. Taking into consideration this production under seawater-like salinity, and the identification of main key players involved in the carbon fluxes, this work paves the way for a feasible production of this, or other compatible solutes, using optimized Synechocystis chassis in a pilot-scale.

Two field experiments were carried out during 2018–2019 at the Hamedan and Qom sites, which are different ecological locations of Iran. Hamedan was selected as a temperate climate and Qom as semi-arid to study some of the morphological and physiological traits of fodder beet in various climatic conditions. The experimental map was designed as a split–split-plot in the form of a randomized complete block with three replications. The main plots were assigned three levels of methanol (i.e., control, 15, and 30% by volume). The sub-plots were restricted to two glycine betaine (GB) concentrations (i.e., control and four grams per liter of methanol consumed, and the sub-sub-plots consisted of six different genotypes of fodder beet). The results showed that root length, root and foliage yield/ha, as well as sugar content increased with the incremental level of methanol and/or GB concentration. Foliar application of methanol and GB also significantly changed the qualitative parameters including sodium content, catalase value, crude protein percentage, and dry matter digestibility. It should be noted that two sites differed in most of the investigated traits, so that Hamedan treatment had a more active role on fodder beet growth, yield, and quality compared with Qom. Meanwhile, there was a significant difference between fodder beet genotypes. Generally, spraying fodder beet with 15%by volume methanol or GB with concentration of four grams per liter of methanol is the recommended treatment for raising forage yield under the ecological circumstances of this research.

BackgroundToday, stress moderators are employed to mitigate crop loss due to the adverse effects of environmental stress. The current research aimed to investigate the impacts of time and stress moderator types on agro-physiological responses of cotton on conventional and double-cropping systems during 2017 and 2018 under saline conditions. A split-plot factorial experiment was carried out in a randomized complete block design with three replications. Cultivation system [conventional (recommended planting date) and double-cropping systems (sowing after harvest of wheat)] were considered as the main plots, and stress moderator type at four levels [water control, 2 mmol·L–1Salicylic acid (SA), 100 mmol·L–1Glycine betaine (GB), and 100 μmol·L–1sodium nitroprusside (SNP)] and application time (flowering and flowering + bolling stages) were regarded as subplots.ResultsPlant height, reproductive branch number, the number of bolls, 10-boll weight, 1 000-seed weight, biological yield, seed cotton yield, lint yield, chlorophyll a, chlorophyll b, carotenoids, total chlorophyll, sodium, potassium, and proline content were less in double-cropping system comparing with conventional system. Spraying with the stress moderators alleviated soil salinity effects on yield, yield components, and biochemical traits of cotton. SNP spraying led to maximum plant height, branch number, the number of bolls per plant, 10-boll weight and seed cotton yield. SA spraying yielded the highest 1 000-seed weight, biological yield, lint percentage and lint yield. The highest chlorophyll a, chlorophyll b, carotenoids, and total chlorophyll content resulted from SNP spraying. Yield, yield components, and biochemical traits did not respond to the stress moderator types in double-cropping system. However, the highest chlorophyll a, carotenoids, proline content, the number of bolls per plant, and seed cotton yield resulted from SNP spraying in conventional system. No statistically significant differences were observed between spraying with SNP and SA in most studied traits.ConclusionsThe results suggest that the optimum cotton planting time and SNP spraying could be recommended for producing the most suitable yield under saline conditions.HighlightsExternal application of stress modulators increases salinity stress tolerance.Spraying with sodium nitroprusside has more moderating effect.Agro-physiological response of cotton to moderators is stronger in early sowing.Maximum seed cotton yield was achieved at early sowing and spraying sodium nitroprusside.Delayed cultivation reduces cotton yield.

Osmotic stress is a major physiologic dysfunction that alters the water movement across the cell membrane. Soil salinity and water stress are major causal factors of osmotic stress that severely affect agricultural productivity and sustainability. Herein, we suggested and evaluated the impact of integrated biochar-based soil amendment and exogenous glycine betaine application on the growth, physiology, productivity, grain quality, and osmotic stress tolerance of rice (Oryza sativa L., cv. Sakha 105) grown in salt-affected soil under three irrigation intervals (6, 9, or 12 days), as well as soil properties and nutrient uptake under field conditions during the 2019 and 2020 seasons. Our findings showed that dual application of biochar and glycine betaine (biochar + glycine betaine) reduced the soil pH, electrical conductivity, and exchangeable sodium percentage. However, it enhanced the K+ uptake which increased in the leaves of treated-rice plants. Additionally, biochar and glycine betaine supplementation enhanced the photosynthetic pigments (chlorophyll a, b, and carotenoids) and physiological attributes (net photosynthetic rate, stomatal conductance, relative water content, and electrolyte leakage) of osmotic-stressed rice plants. Biochar + glycine betaine altered the activity of antioxidant-related enzymes (catalase, ascorbate peroxide, and peroxidase). Moreover, it improved the yield components, biological yield, and harvest index, as well as the nutrient value of rice grains of osmotic-stressed rice plants. Collectively, these findings underline the potential application of biochar and glycine betaine as a sustainable eco-friendly strategy to improve plant resilience, not only rice, but other plant species in general and other cereal crops in particular, to abiotic stress, particularly those growing in salt-affected soil.

Cold seeps are globally widespread seafloor ecosystems that feature abundant methane production and flourishing chemotrophic benthic communities. Chemical evidence indicates that cold seep methane is largely biogenic; however, the primary methane-producing organisms and associated pathways involved in methanogenesis remain elusive. This work detected methane production when glycine betaine (GBT) or trimethylamine (TMA) was added to the sediment microcosms of the Formosa cold seep, South China Sea. The methane production was suppressed by antibiotic inhibition of bacteria, while GBT was accumulated. This suggests that the widely used osmoprotectant GBT could be converted to cold seep biogenic methane via the synergistic activity of bacteria and methanogenic archaea because archaea are not sensitive to antibiotics and no bacteria are known to produce ample methane (mM). 16S rRNA gene diversity analyses revealed that the predominant bacterial and archaeal genera in the GBT-amended methanogenic microcosms included Oceanirhabdus and Methanococcoides. Moreover, metagenomic analyses detected the presence of grdH and mtgB genes that are involved in GBT reduction and demethylation, respectively. Two novel species were obtained, including bacterium Oceanirhabdus seepicola, which reduces GBT to TMA, and a methanogenic archaeon, Methanococcoides seepicolus, which produces methane from TMA and GBT. The two strains reconstituted coculture efficiently converted GBT to methane at 18°C; however, at 4°C addition of dimethylglycine (DMG), the GBT demethylation product, was necessary. Therefore, this work demonstrated that GBT is the precursor not only of the biogenic methane but also of the cryoprotectant DMG to the microorganisms at the Formosa cold seep. IMPORTANCE Numerous cold seeps have been found in global continental margins where methane is enriched in pore waters that are forced upward from sediments. Therefore, high concerns have been focused on the methane-producing organisms and the metabolic pathways in these environments because methane is a potent greenhouse gas. In this study, GBT was identified as the main precursor for methane in the Formosa cold seep of the South China Sea. Further, synergism of bacteria and methanogenic archaea was identified in GBT conversion to methane via the GBT reduction pathway, while methanogen-mediated GBT demethylation to methane was also observed. In addition, GBT-demethylated product dimethyl glycine acted as a cryoprotectant that promoted the cold seep microorganisms at cold temperatures. GBT is an osmoprotectant that is widely used by marine organisms, and therefore, the GBT-derived methanogenic pathway reported here could be widely distributed among global cold seep environments.

Staphylococcus aureus is an opportunistic pathogen that can cause soft tissue infections but is also a frequent cause of foodborne illnesses. One contributing factor for this food association is its high salt tolerance allowing this organism to survive commonly used food preservation methods. How this resistance is mediated is poorly understood, particularly during long‐term exposure. In this study, we used transposon sequencing (TN‐seq) to understand how the responses to osmotic stressors differ. Our results revealed distinctly different long‐term responses to NaCl, KCl and sucrose stresses. In addition, we identified the DUF2538 domain containing gene SAUSA300_0957 (gene 957) as essential under salt stress. Interestingly, a 957 mutant was less susceptible to oxacillin and showed increased peptidoglycan crosslinking. The salt sensitivity phenotype could be suppressed by amino acid substitutions in the transglycosylase domain of the penicillin‐binding protein Pbp2, and these changes restored the peptidoglycan crosslinking to WT levels. These results indicate that increased crosslinking of the peptidoglycan polymer can be detrimental and highlight a critical role of the bacterial cell wall for osmotic stress resistance. This study will serve as a starting point for future research on osmotic stress response and help develop better strategies to tackle foodborne staphylococcal infections., Staphylococcus aureus is able to grow in the presence of high concentrations of NaCl but the exact genetic factors contributing to this are unknown. Using a high‐throughput TN‐seq approach, we identified gene 957 as an important factor for the salt stress resistance in S. aureus. A 957‐mutant was not only salt sensitive but also showed increased peptidoglycan crosslinking, altogether highlighting the cell wall as an important barrier against osmotic stress.

Made available in DSpace on 2019-10-04T12:37:51Z (GMT). No. of bitstreams: 0 Previous issue date: 2019-06-01 Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES) CSIRO, Sandwich PhD scholarship Australian Research Council Centre of Excellence in Plant Energy Biology Sugarcane is an important crop in tropical regions of the world, often being exposed to environments with high salinity, but little is known of genetic variation in salt tolerance. The aim of this work was to compare the performance of two genetically diverse cultivars of sugarcane under different concentrations of salinity (0, 40, 80 and 160 mM NaCl) over a period of 30 days. SP 81-3250 was more salt-tolerant and maintained its rate of biomass production, photosynthesis and leaf area up to 160 mM NaCl, whereas IAC 87-3396 was sensitive to 80 mM NaCl. SP 81-3250 maintained very low concentrations of Na+ in both leaves and roots with increasing time and salinity, whereas in IAC 87-3396 the Na+ concentrations were 2-5 times higher. SP 81-3250 had a greater accumulation of proline, and lower lipid peroxidation, whereas glycine betaine and sucrose concentrations were similar in the two cultivars. This suggests that the tolerance of SP 81-3250 to high salinity was due to its ability to exclude Na+ while taking up water from the soil, and that measurements of Na+ concentration in leaves could be used to select salt-tolerant genotypes for saline areas. Univ Estadual Paulista, Dept Biol Aplicada Agr, BR-14884900 Jaboticabal, SP, Brazil CSIRO Agr & Food, GPO Box 1700, Canberra, ACT 2601, Australia Univ Western Australia, Sch Agr & Environm, Crawley, WA 6009, Australia Univ Western Australia, ARC Ctr Excellence Plant Energy Biol, Crawley, WA 6009, Australia Univ Estadual Paulista, Dept Biol Aplicada Agr, BR-14884900 Jaboticabal, SP, Brazil CSIRO, Sandwich PhD scholarship: BEX12786/13-6 Australian Research Council Centre of Excellence in Plant Energy Biology: CE140100008

Despite being protected, the number of plants that belong toSideritisgenus has decreased in the last years due to the urbanization of its natural habitat.Sideritis raeseriBoiss and Heldr. subsp.raeseriknown as mountain tea of Parnassus or Velouchi is a valuable genetic resource of the Greek flora because of its special characteristics, so, the development of a micropropagation protocol is crucial to its conservation and use in breeding programs. The objectives of the study were to evaluate the effects of glycine betaine (GB), polyvinylpyrrolidone (PVP-10) and D-mannitol applied in different concentrations in combination with auxins [α-naphthaleneacetic acid (NAA), indole-3-butyric acid (IBA)] on shoot proliferation and rooting of shoot-tip explants of the studied species. Better shoot proliferation results were obtained in the following two combination treatments: (1) 100 mg/l GB + 0.5 mg/l NAA (5.5 shoots/explant 2.34 cm long, 75% shoot multiplication), and (2) 2.5 g/l D-mannitol + 3 mg/l IBA (2.92 shoot/explant 83.33% shoot multiplication percentage). In relation to rooting, better results were performed only with PVP at 0.5 g/l applied simultaneously with 0.5 mg/l NAA (12 roots/rooted microcutting, 1.47 cm root length, 90% rooting). Theex vitrosurvival percentage of rooted microcuttings derived from all 5 experiments was ranged between 68% and 93%.

The production and consumption of organic matter by marine organisms plays a central role in the marine carbon cycle. Labile organic compounds (metabolites) are the major currency of energetic demands and organismal interaction, but these compounds remain elusive because of their rapid turnover and concomitant minuscule concentrations in the dissolved organic matter pool. Organic osmolytes are a group of small metabolites synthesized at high intracellular concentrations (mM) to regulate cellular osmolarity and have the potential to be released as abundant dissolved substrates. Osmolytes may represent an essential currency of exchange among heterotrophic prokaryotes and primary and secondary producers in marine food webs. For example, the well-known metabolite dimethylsulfoniopropionate (DMSP) is used as an osmolyte by some phytoplankton and can be subsequently metabolized by 60% of the marine bacterial community, supplying up to 13% of the bacterial carbon demand and 100% of the bacterial sulfur demand. While marine osmolytes have been studied for decades, our understanding of their cycling and significance within microbial communities is still far from comprehensive. Here, we surveyed the genes responsible for synthesis, breakdown, and transport of 14 key osmolytes. We systematically searched for these genes across marine bacterial genomes (n = 897) and protistan transcriptomes (n = 652) using homologous protein profiles to investigate the potential for osmolyte metabolisms. Using the pattern of gene presence and absence, we infer the metabolic potential of surveyed microbes to interact with each osmolyte. Specifically, we identify: (1) complete pathways for osmolyte synthesis in both prokaryotic and eukaryotic marine microbes, (2) microbes capable of transporting osmolytes but lacking complete synthesis and/or breakdown pathways, and (3) osmolytes whose synthesis and/or breakdown appears to be specialized and is limited to a subset of organisms. The analysis clearly demonstrates that the marine microbial loop has the genetic potential to actively recycle osmolytes and that this abundant group of small metabolites may function as a significant source of nutrients through exchange among diverse microbial groups that significantly contribute to the cycling of labile carbon.

Abiotic stresses, mainly salinity and drought, are the most important environmental threats that constrain worldwide food security by hampering plant growth and productivity. Plants cope with the adverse effects of these stresses by implementing a series of morpho-physio-biochemical adaptation mechanisms. Accumulating effective osmo-protectants, such as proline and glycine betaine (GB), is one of the important plant stress tolerance strategies. These osmolytes can trigger plant stress tolerance mechanisms, which include stress signal transduction, activating resistance genes, increasing levels of enzymatic and non-enzymatic antioxidants, protecting cell osmotic pressure, enhancing cell membrane integrity, as well as protecting their photosynthetic apparatus, especially the photosystem II (PSII) complex. Genetic engineering, as one of the most important plant biotechnology methods, helps to expedite the development of stress-tolerant plants by introducing the key tolerance genes involved in the biosynthetic pathways of osmolytes into plants. Betaine aldehyde dehydrogenase (BADH) is one of the important genes involved in the biosynthetic pathway of GB, and its introduction has led to an increased tolerance to a variety of abiotic stresses in different plant species. Replacing down-regulated ferredoxin at the acceptor side of photosystem I (PSI) with its isofunctional counterpart electron carrier (flavodoxin) is another applicable strategy to strengthen the photosynthetic apparatus of plants under stressful conditions. Heterologous expression of microbially-sourced flavodoxin (Fld) in higher plants compensates for the deficiency of ferredoxin expression and enhances their stress tolerance. BADH and Fld are multifunctional transgenes that increase the stress tolerance of different plant species and maintain their production under stressful situations by protecting and enhancing their photosynthetic apparatus. In addition to increasing stress tolerance, both BADH and Fld genes can improve the productivity, symbiotic performance, and longevity of plants. Because of the multigenic and complex nature of abiotic stresses, the concomitant delivery of BADH and Fld transgenes can lead to more satisfying results in desired plants, as these two genes enhance plant stress tolerance through different mechanisms, and their cumulative effect can be much more beneficial than their individual ones. The importance of BADH and Fld genes in enhancing plant productivity under stress conditions has been discussed in detail in the present review.

Background Glycine betaine (GB) plays a crucial role in plants responding to abiotic stresses. Studying the physiological response of cotton seedlings to exogenous GB under salt stress provides a reference for the application of GB to improve the resistance of cotton seedlings under salt stress. The purpose of this research is to examine the impacts of foliar-applied GB on leaf stomatal structure and characteristics, gas exchange and chlorophyll fluorescence characteristics and plant growth indicators of Gossypium hirsutum L. under NaCl stress conditions. Results Under the salinity of 150 mM, the four concentrations of GB are 0, 2.5, 5, and 7.5 mM, and the control (CK) was GB-untreated non-saline. Salt stress negatively affected leaf stomata as well as gas exchange and chlorophyll fluorescence and decreased plant growth parameters of cotton seedlings. The treatment with 5 mM GB significantly increased the evolution of photosynthetic rate (Pn), transpiration rate (Tr), intracellular CO2 concentration (Ci) and stomatal conductance (gs) compared to the GB-untreated saline treatment. The Exogenous foliar-applied GB has sustainably decreased the carboxylation efficiency (Pn/Ci) and water use efficiency (WUE). The concentration of 5 mM GB leads to a significant improvement of leaf stomatal characteristics. The leaf gas exchange attributes correlated positively with stomatal density (SD), stomatal length (SL) and stomatal with (SW). Conclusion The overall results suggested that exogenous foliar supplementation with GB can effectively alleviate the damage of salt stress to cotton seedlings. The effect of applying 5 mM GB could be an optional choice for protecting cotton seedlings from NaCl stress through promoting the stomatal functions, photosynthetic activities and growth characteristics.

Salinity is one of the most serious factors limiting plant growth which can provoke significant losses in agricultural crop production, particularly in arid and semi-arid areas. This study aimed to investigate whether melatonin (MT; 0.05 and 0.1 mM), which has pleiotropic roles, has a better effect than glycine betaine (GB; 10 and 50 mM) on providing salt tolerance in a woody plant Dalbergia odorifera T. Chen. Also, the alternative oxidase activity (AOX) in plant subjected to MT or GB under salinity (150 and 250 mM) was evaluated given that the effect of exogenous MT or GB on AOX has not been reported yet. The results showed that the exogenous application of GB on the seedlings of D. odorifera increased the plant growth parameters, relative water content, total of chlorophyll content, and carotenoid content compared with well-watered and MT treatments. Under severe salinity, the seedlings subjected to GB showed, a significant enhancement in water use efficiency, transpiration, and net photosynthetic rate regardless to MT-treated seedlings. The levels of proline and soluble sugar in the seedlings treated with MT or GB decreased significantly under mild and severe salinity correlated with those in salt-stressed seedlings. Furthermore, GB-treated plants exhibited a significant inhibition of malondialdehyde content compared with MT-treated plants. The concentration of thiols and phenolic compounds were significantly enhanced in the leaves of seedlings treated with MT compared with those treated with GB. Under salt stress condition, GB scavenged significantly higher levels of hydrogen peroxide than MT; while under severe salinity, plants subjected to MT showed better scavenging ability for hydroxyl radicals compared with GB-treated seedlings. The results demonstrated also an enhancement of the levels of superoxide dismutase (SOD), guaiacol peroxidase, and AOX activities in seedlings treated with GB or MT compared with salt-stressed plants. The catalase activity (CAT) was increased by 0.05 mM MT and 0.1 mM GB under mild salinity. Meanwhile, the AOX activity under severe salinity was enhanced only by GB 50 mM. The findings of this study suggested that GB-treated seedlings possessed a better salt tolerance in comparison with MT-treated seedlings.

Soil salinization adversely affects agricultural productivity. Mitigating the adverse effects of salinity represents a current major challenge for agricultural researchers worldwide. The effects of exogenously applied glycine betaine (GB) and salicylic acid (SA) on mitigating sodium toxicity and improving the growth of cotton seedlings subjected to salt stress remain unclear. The treatments in a phytotron included a control (CK, exogenously untreated, non-saline), two NaCl conditions (0 and 150 mM), four exogenous GB concentrations (0, 2.5, 5.0, and 7.5 mM), and four exogenous SA concentrations (0, 1.0, 1.5, and 2.0 mM). The shoot and roots exposed to 150 mM NaCl without supplementation had significantly higher Na+ and reduced K+, Ca2+, and Mg2+ contents, along with lowered biomass, compared with those of CK. Under NaCl stress, exogenous GB and SA at all concentrations substantially inversed these trends by improving ion uptake regulation and biomass accumulation compared with NaCl stress alone. Supplementation with 5.0 mM GB and with 1.0 mM SA under NaCl stress were the most effective conditions for mitigating Na+ toxicity and enhancing biomass accumulation. NaCl stress had a negative effect on plant growth parameters, including plant height, leaf area, leaf water potential, and total nitrogen (N) in the shoot and roots, which were improved by supplementation with 5.0 mM GB or 1.0 mM SA. Supplementation with 5.0 mM exogenous GB was more effective in controlling the percentage loss of conductivity (PLC) under NaCl stress.

In addition to their haloalkaline conditions, soda lakes can also harbor a variety of other extreme parameters, to which their microbial communities need to adapt. However, for most of these supplementary stressors, it is not well known yet how haloalkaliphiles adapt and resist., The genus Thioalkalivibrio comprises sulfur-oxidizing bacteria thriving in soda lakes at high pH and salinity. Depending on the geographical location and the season, these lakes can strongly vary in temperature. To obtain a comprehensive understanding of the molecular and physiological adaptations to low temperature, we compared the responses of two Thioalkalivibrio strains to low (10°C) and high (30°C) temperatures. For this, the strains were grown under controlled conditions in chemostats and analyzed for their gene expression (RNA sequencing [RNA-Seq]), membrane lipid composition, and glycine betaine content. The strain Thioalkalivibrio versutus AL2T originated from a soda lake in southeast Siberia that is exposed to strong seasonal temperature differences, including freezing winters, whereas Thioalkalivibrio nitratis ALJ2 was isolated from an East African Rift Valley soda lake with a constant warm temperature the year round. The strain AL2T grew faster than ALJ2 at 10°C, likely due to its 3-fold-higher concentration of the osmolyte glycine betaine. Moreover, significant changes in the membrane lipid composition were observed for both strains, leading to an increase in their unsaturated fatty acid content via the Fab pathway to avoid membrane stiffness. Genes for the transcriptional and translational machinery, as well as for counteracting cold-induced hampering of nucleotides and proteins, were upregulated. Oxidative stress was reduced by induction of vitamin B12 biosynthesis genes, and growth at 10°C provoked downregulation of genes involved in the second half of the sulfur oxidation pathway. Genes for intracellular signal transduction were differentially expressed, and interestingly, AL2T upregulated flagellin expression, whereas ALJ2 downregulated it. IMPORTANCE In addition to their haloalkaline conditions, soda lakes can also harbor a variety of other extreme parameters, to which their microbial communities need to adapt. However, for most of these supplementary stressors, it is not well known yet how haloalkaliphiles adapt and resist. Here, we studied the strategy for adaptation to low temperature in the haloalkaliphilic genus Thioalkalivibrio by using two strains isolated from soda lakes with different temperature regimes. Even though the strains showed a strong difference in growth rate at 10°C, they exhibited similar molecular and physiological adaptation responses. We hypothesize that they take advantage of resistance mechanisms against other stressors commonly found in soda lakes, which are therefore maintained in the bacteria living in the absence of low-temperature pressure. A major difference, however, was detected for their glycine betaine content at 10°C, highlighting the power of this osmolyte to also act as a key compound in cryoprotection.

Several inorganic and organic compounds including glycine betaine (GB) are presently being used as an exogenous application to enhance tolerance in plants to different environmental stresses. The current study assessed to what extent exogenously applied GB could improve the gaseous exchange capacity and primary and secondary metabolites in two accessions (16178 and 16180) of safflower (Carthamus tinctorius L.) plants under drought stress. Three-week-old plants of both safflower accessions were subjected to well-watered (control) or water-deficit conditions (60% field capacity (FC)). Three levels of GB (control, 50 mM and 100 mM) were sprayed to the foliage of the control and stressed plants after one month of drought application. After two weeks of foliar application of GB, gas exchange characteristics and other biochemical parameters were determined. The results showed that water deficiency markedly suppressed plant biomass, chlorophyll contents, photosynthesis rate (A), water use efficiency (A/E), stomatal conductance (gs) and relative water contents (RWC) of both accessions of safflower, while it enhanced the levels of osmolytes (GB and proline), hydrogen peroxide (H2O2) and total phenolics. Foliar application of GB was effective in enhancing the plant biomass, chlorophyll contents, gs, sub-stomatal CO2 concentration (Ci), Ci/Ca ratio, osmolytes, H2O2, ascorbic acid (AsA), total phenolics and RWC in safflower plants under water shortage. Thus, exogenous application of GB could be used as an effective strategy to improve plant growth, photosynthetic attributes and secondary metabolites in safflower plants under water deficit conditions.

Alexandrium minutum and Alexandrium pacificum are representatives of the dinoflagellate genus that regularly proliferate on the French coasts and other global coastlines. These harmful species may threaten shellfish harvest and human health due to their ability to synthesize neurotoxic alkaloids of the saxitoxin group. However, some dinoflagellates such as A. minutum, and as reported here A. pacificum as well, may also have a beneficial impact on the environment by producing dimethylsulfoniopropionate-DMSP, the precursor of dimethylsulfur-DMS and sulfate aerosols involved in climate balance. However, environmental conditions might influence Alexandrium physiology towards the production of harmful or environmentally friendly compounds. After assessing the influence of two salinity regimes (33 and 38) relative to each species origin (Atlantic French coast and Mediterranean Lagoon respectively), it appears that DMSP and toxin content was variable between the three experimented strains and that higher salinity disadvantages toxin production and tends to favor the production of the osmolytes DMSP and glycine betaine. Hence, this key metabolite production is strain and species-dependent and is influenced by environmental conditions of salinity which in turn, can diversely affect the environment. Widespread coastal blooms of A. minutum and A. pacificum, although being a risk for seafood contamination with toxins, are also a DMSP and DMS source that potentially contribute to the ecosystem structuration and climate. Regarding recent advances in DMSP biosynthesis pathway, 3 dsyB homologs were found in A. minutum but no homolog of the diatom sequence TpMMT.

Background Chromium is the most toxic pollutant that negatively affects a plant’s metabolic activities and yield. It reduces plant growth by influencing the antioxidant defence system’s activities. In the present study, a completely randomized block design experiment with three plants/pot in three replication was conducted on three varieties of sorghum viz. SSG 59–3, HJ 513 (multi-cut) and HJ 541 (single-cut) for amelioration of chromium toxicity (2 & 4 ppm) by exogenous application of GB (50 & 100 mM) with and without AMF in soil. The ameliorative effects were tested at two growth stages viz. vegetative (35 DAS) and grain filling (95 DAS), in terms of Cr uptake, grain yield, antioxidative defence system parameters (viz. enzymes – SOD, APX, CAT, GR, POX and metabolites – proline, glutathione, ascorbate, β-carotene) and indices of oxidative stress parameters (viz. PPO, H2O2, and MDA). Results The results delineated that Cr uptake and indices of oxidative stress were increased with increasing concentration of Cr stress in all the varieties (HJ 541, HJ513 & SSG 59–3) at both the growth stages (35 & 95 DAS). At higher concentration (4 ppm), Cr stress decreased the grain yield (45–50%) as compared with controls. Polyphenoloxidase activity, MDA and H2O2 content increased at both growth stages in all the varieties. However, antioxidative enzymes and metabolite activities increased due to Cr stress but this increase was not sufficient to counteract with ROS generated under Cr stress which was enhanced on the application of AMF and GB either individually or in combination (spiked in soil). It decreased the indices of oxidative stress and ameliorated the Cr toxicity and increased grain yield (65–70%) in all the varieties. Conclusions Both GB and AMF improved the antioxidative activities and stress tolerance capacity of the plant. Glycine betaine at both 50 and 100 mM level, significantly ameliorated Cr toxicity. However, AMF concomitantly with GB further boosts up the amelioration behaviour of the plant against Cr toxicity, at both growth stages in all the varieties. The combination of 100 mM GB with 10 g AMF was observed most effective among all the treatments. Among the varieties, SSG 59–3 had the lowest chromium uptake, indices of oxidative stress, and highest antioxidative system’s activity as compared to HJ 513 followed by HJ 541 variety. Thus AMF and GB either individually or in combination may be used to maintain plant yield attributes under Cr toxicity.

International audience; The solubility of hydrophobic betainium bis(trifluoromethylsulfonyl)imide ionic liquid in an aqueous phase with controlled acidity has been investigated by a standard liquid-liquid extraction technique. The effect of zwitterionic betaine added into the aqueous phase has also been studied. Quantitative NMR was employed to characterize the distribution behavior of the ionic liquid constituents between the two phases. A mathematical model to describe the ionic liquid solubility in the aqueous phase was developed. This approach was utilized to estimate the conditional solubility product constant of the ionic liquid. The possibility of the transfer of some dissociation products back into the organic phase was investigated. (C) 2018 Elsevier B.V. All rights reserved.

Made available in DSpace on 2021-06-25T12:23:36Z (GMT). No. of bitstreams: 0 Previous issue date: 2020-01-01. Added 1 bitstream(s) on 2021-07-15T15:24:26Z : No. of bitstreams: 1 S1807-86212020000105019.pdf: 2036779 bytes, checksum: 670a5d5ff8fdb66581eea2117111bc94 (MD5) Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq) Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES) The present study aims to determine whether exogenous salicylic acid (SA) or spermidine (Spd) has any protective effect against salt stress. Seeds were subjected to 0, 20, 40, and 60 mM NaCl with or without salicylic acid or spermidine (0.5 mM) for 10 days. The evaluated variables were germination rate, shoot and root dry masses, glycine betaine content, lipid peroxidation, and the activities of superoxide dismutase (SOD), catalase (CAT), and ascorbate peroxidase (APX). The data were subjected to Tukey's test (p

A study was carried out to evaluate the effectiveness of sugar beet extract (SBE) and glycine betaine (GB) in mitigating the adverse effects of drought stress on two maize cultivars. Seeds (caryopses) of two maize cultivars, Sadaf (drought-tolerant) and Sultan (drought-sensitive) were sown in plastic pots. Plants were subjected to different (100%, 75% and 60% field capacity (FC)) water regimes. Then, different levels of SBE (3% and 4%) and GB (3.65 and 3.84 g/L) were applied as a foliar spray after 30 days of water deficit stress. Drought stress significantly decreased plant growth and yield attributes, chlorophyll pigments, while it increased relative membrane permeability (RMP), levels of osmolytes (GB and proline), malondialdehyde (MDA), total phenolics and ascorbic acid as well as the activities of superoxide dismutase (SOD) and peroxidase (POD) enzymes in both maize cultivars. Exogenous application via foliar spray with SBR or GB improved plant growth and yield attributes, chlorophyll pigments, osmolyte concentration, total phenolics, ascorbic acid and the activities of reactive oxygen species (ROS) scavenging enzymes (SOD, POD and catalase; CAT), but reduced leaf RMP and MDA concentration. The results obtained in this study exhibit the role of foliar-applied biostimulants (natural and synthetic compounds) in enhancing the growth and yield of maize cultivars by upregulating the oxidative defense system and osmoprotectant accumulation under water deficit conditions.

The aim of the research was to test the effect of foliar-applied solutions of ascorbic acid (AsA), glycine betaine (GB), salicylic acid (SA) and water as a second control on the yield and seed germination of the soybean (Glycine hispida (Moench) Maxim.) cultivar 'Sava' as well as their influence on the content of N, P, K and Ca in soybean seeds, and to estimate the intensity of lipid peroxidation and free proline content in soybean seedlings. The plants were treated with the solutions twice. The 1st spray treatment was done at growth stage 6: flowering (main shoot), when first flowers opened (sporadically in population), and the 2nd spray treatment was done at growth stage 7: development of fruits and seeds, when the first pod had reached final length of 15-20 mm. Foliar application of the solutions had a significant impact on seed yield and seed germination in both years of the experiment. In a favourable year 2016, treatments with ascorbic acid, glycine betaine and salicylic acid solution significantly increased seed yield by 6.21-9.67%. However, in an unfavourable year 2017, the effect was significantly greater with the application of water (yield increase 6.48%), while the application of glycine betaine solution even reduced the seed yield. In a favourable year 2016, treatments with ascorbic acid and salicylic acid solution significantly increased seed germination by 7% and 8%, respectively. In an unfavourable year 2017, also only ascorbic acid and salicylic acid significantly increased seed germination by 11% and 9%, respectively. However, when using cold test, when the seed was exposed to a low temperature of 10 degrees C for 7 days and then placed under optimal seed germination conditions for 4 days, ascorbic acid and glycine betaine solution reduced seed germination in both experimental years. Therefore, further research is needed on the methods designed to increase seed germination of soybean, both in favourable and adverse conditions in Serbia.

Recent studies indicate that environmentally abundant quaternary amines (QAs) are a primary source for methanogenesis, yet the catabolic enzymes are unknown. We hypothesized that the methanogenic archaeon Methanolobus vulcani B1d metabolizes glycine betaine (GB) through a corrinoid-dependent GB:coenzyme M (CoM) methyl transfer pathway. The draft genome sequence of M. vulcani B1d revealed a gene encoding a predicted non-pyrrolysine MttB homolog (MV8460) with high sequence similarity to the GB methyltransferase encoded by Desulfitobacterium hafniense Y51. MV8460 catalyzes GB-dependent methylation of free cob(I)alamin indicating it is an authentic MtgB enzyme. Proteomic analysis revealed that MV8460 and a corrinoid binding protein (MV8465) were highly abundant when M. vulcani B1d was grown on GB relative to growth on trimethylamine. The abundance of a corrinoid reductive activation enzyme (MV10335) and a methylcorrinoid:CoM methyltransferase (MV10360) were significantly higher in GB-grown B1d lysates compared to other homologs. The GB:CoM pathway was fully reconstituted in vitro using recombinant MV8460, MV8465, MV10335, and MV10360. Demonstration of the complete GB:CoM pathway expands the knowledge of direct QA-dependent methylotrophy and establishes a model to identify additional ecologically relevant anaerobic quaternary amine pathways.

International audience; Dinophysis is the main dinoflagellate genus responsible for diarrheic shellfish poisoning (DSP) in human consumers of filter feeding bivalves contaminated with lipophilic diarrheic toxins. Species of this genus have a worldwide distribution driven by environmental conditions (temperature, irradiance, salinity, nutrients etc.), and these factors are sensitive to climate change. The D. acuminata-complex may contain several species, including D. sacculus. The latter has been found in estuaries and semi-enclosed areas, water bodies subjected to quick salinity variations and its natural repartition suggests some tolerance to salinity changes. However, the response of strains of D. acuminata-complex (D. cf. sacculus) subjected to salinity stress and the underlying mechanisms have never been studied in the laboratory. Here, a 24 h hypoosmotic (25) and hyperosmotic (42) stress was performed in vitro in a metabolomic study carried out with three cultivated strains of D. cf. sacculus isolated from the French Atlantic and Mediterranean coasts. Growth rate, biovolume and osmolyte (proline, glycine betaine and dimethylsulfoniopropionate (DMSP)) and toxin contents were measured. Osmolyte contents were higher at the highest salinity, but only a significant increase in glycine betaine was observed between the control (35) and the hyperosmotic treatment. Metabolomics revealed significant and strain-dependent differences in metabolite profiles for different salinities. These results, as well as the absence of effects on growth rate, biovolume, okadaic acid (OA) and pectenotoxin (PTXs) cellular contents, suggest that the D. cf. sacculus strains studied are highly tolerant to salinity variations.

Safflower ( Carthamus tinctorius L.) is an important oilseed crop, usually grown on a small scale and in salt-affected soils. Salinity stress can cause oxidative damage to plants. Upregulation of the antioxidant defense system induced by glycine betaine (GlyBet) alleviates the damaging effects of oxidative stress in plants. In the present investigation, seeds were treated with 0, 10, 30 and 60 mM of GlyBet solutions. Germination and the primary growth of the seedling were examined using sodium chloride salt (NaCl) at 0 (non-stress), 50, 100 and 150 mM concentrations. The obtained results indicate that at 50 and 100 mM NaCl, priming with 30 and 60 mM GlyBet increased root and shoot lengths compared to the control (0 mM). In addition, at all stress levels, priming with 60 mM GlyBet led to lower malondialdehyde, total soluble sugars and proline contents than in control seedlings. Priming with GlyBet increased catalase (CAT), superoxide dismutase (SOD) enzyme activities and protein content, while it reduced the activity of peroxidase (POD) under salinity stress. In addition, priming with GlyBet reduced the Na + /K + ratio of seedlings and increased K + under all salinity stress levels. Priming with 60 mM GlyBet also reduced the Na + content under 150 mM NaCl. Together, these results show that 60 mM GlyBet had the most pronounced effect on tolerance to salinity stress in safflower seedling. The GlyBet-increased tolerance to salt in safflower was mainly related to increased CAT and SOD activities, and the prevention of cell membrane damage as a result of reduced lipid peroxidation and improved ion homeostasis under salinity stress condition. DOI: 10.2298/ABS160216089A Received: February 16, 2016; Revised: April 25, 2016; Accepted: May 23, 2016; Published online: October 21, 2016 How to cite this article: Alasvandyari F, Mahdavi B, Hosseini Shahab M. Glycine betaine affects the antioxidant system and ion accumulation and reduces salinity-induced damage in safflower seedlings. Arch Biol Sci. 2017;69(1):139-47.

Pot experiments in sandy soil were carried out to investigate foliar applications of [selenium (5 & 10 μM Se), glycine betaine (5 & 10 mM GB) and seaweed extract (2 & 4% SWD)] on cowpea plant, under salt stress (0 & 50 mM NaCl). Growth, yield parameters and biochemical constituents were determined. The obtained results indicated that all foliar treatments under both salinity concentrations achieved an increment in most of the growth, yield parameters and biochemical constituents. As for the effect of single treatment on growth and yield parameters, 5 μM Se without salinity revealed higher significant values than all treatments followed by 10 μM Se under 50 mM NaCl. Meanwhile, 4% SWE under 50 mM NaCl showed the highest values of photosynthetic pigments, proline and Phenylalanine ammonia lyase (PAL). On the other hand, 10 mM GB 50 mM under NaCl recorded the highest value of total soluble sugars (TSS), while a significant increase in protein concentrations was observed by 5 μM Se without salinity.

Snap bean is a salt sensitive plant and suffers from losses in yield and pod quality with any little increase of salt concentration in irrigation water. In order to study the effect of salicylic acid (SA), spermidine (Spd), and glycine betaine (GB) as phytoprotectants on enhancing growth, yield and pod quality of snap bean under different levels of NaCl salinity, an outdoor pot experiment was conducted in 2012 and 2013 seasons. Salinity was applied as NaCl form at 0 and 2000 ppm. The concentrations of foliar treatments were two levels for each treatment; the first level was 0 mM which served as control, and the second level was 1 mM SA, 0.5 mM Spd and 5 mM GB, in addition to their combinations. NaCl salinity at 2000 ppm reduced most of vegetative growth parameters such as plant f.w., leaf area ratio and leaf area index, which in turn reflected on the reduction of pods no./plant and yield f.w./plant and an increase in the fruit abscission percentage. Pod moisture % decreased under 2000 ppm NaCl which reduced pod f.w., and increased pod curvature %. Under 2000 ppm NaCl, GB at 5 mM and all its combinations increased membrane stability index, total soluble sugars and total soluble proteins concentration, while reducing free amino acids concentration, which were concomitant with decreasing pod curvature %. Meanwhile, application of SA at 1 mM, GB at 5 mM, and GB5 + SA1 + Spd0.5 increased leaves and pods no./plant, pod moisture %, and pod f.w., which reflected on increasing green pod yield.

8 pages, 3 tables, 1 figure, Glycine betaine (GB) is a natural compound with demonstrated metabolic functions in mammals and beneficial effects on the human body: it protects cells under osmotic stress, as an organic osmolyte, and acts as a methylating agent in several vital biochemical pathways. The main dietary sources of GB are vegetables, such as cereals, spinach, chard and beetroot, above all; shellfish is one of the richest animal sources. A simple method for the quantification of GB in the edible portion of Manila clams (Tapes philippinarum) was setup, using ultra-performance liquid chromatography coupled to tandem mass spectrometry (UHPLC-MS/MS). Isotope-labelled internal standards were used during sample extraction, in order to ensure an accurate quantification. The method was successfully validated following the Commission Decision 657/2002/EC as a guideline; since GB is an endogenous compound, present in very high concentrations in the target matrix, appropriate adjustments were taken. The method developed reported very satisfying performances in terms of linearity, trueness and precision; moreover, its applicability was demonstrated on commercial pools of Manila clams.

“Zhongbai” papaya fruit were treated with 15 mmol/L glycine betaine (GB) and then refrigerated at 6°C for 40 days to study the influence of GB on chilling injury (CI) and possible mechanism associated with energy metabolism. The results exhibited that GB treatment remarkably reduced the CI severity as indicated by lower CI index during storage. GB treatment lowered electrolyte leakage and malondialdehyde content, which accounted for maintenance of membrane integrity and reduced lipid peroxidation. Moreover, GB treatment improved the energy status as revealed by increased adenosine triphosphate (ATP) level, energy charge, and activities of energy metabolism‐related enzymes including mitochondrial membrane H+‐adenosine triphosphatase (H+‐ATPase) and Ca2+‐adenosine triphosphatase (Ca2+‐ATPase), succinate dehydrogenase (SDH), and cytochrome C oxidase (CCO). The results indicate that enhanced chilling tolerance in papaya fruit by GB treatment during cold storage might be ascribed to improved energy status in association with increased activities of energy metabolism‐related enzymes.

Accumulation of compatible solutes is a common stress response of microorganisms challenged by high osmolarity; it can be achieved either through synthesis or import. These processes have been intensively studied in Bacillus subtilis, where systems for the production of the compatible solutes proline and glycine betaine have been identified, and in which five transporters for osmostress protectants (Opu) have been characterized. Glycine betaine synthesis relies on the import of choline via the substrate-restricted OpuB system and the promiscuous OpuC transporter and its subsequent oxidation by the GbsAB enzymes. Transcription of the opuB and gbsAB operons is under control of the MarR-type regulator GbsR, which acts as an intracellular choline-responsive repressor. Modeling studies using the X-ray structure of the Mj223 protein from Methanocaldococcus jannaschii as the template suggest that GbsR is a homo-dimer with an N-terminal DNA-reading head and C-terminal dimerization domain; a flexible linker connects these two domains. In the vicinity of the linker region, an aromatic cage is predicted as the inducer-binding site, whose envisioned architecture resembles that present in choline and glycine betaine substrate-binding proteins of ABC transporters. We used bioinformatics to assess the phylogenomics of GbsR-type proteins and found that they are widely distributed among Bacteria and Archaea. Alignments of GbsR proteins and analysis of the genetic context of the corresponding structural genes allowed their assignment into four sub-groups. In one of these sub-groups of GbsR-type proteins, gbsR-type genes are associated either with OpuA-, OpuB-, or OpuC-type osmostress protectants uptake systems. We focus here on GbsR-type proteins, named OpuAR by us, that control the expression of opuA-type gene clusters. Using such a system from the marine bacterium Bacillus infantis, we show that OpuAR acts as a repressor of opuA transcription, where several compatible solutes (e.g., choline, glycine betaine, proline betaine) serve as its inducers. Site-directed mutagenesis studies allowed a rational improvement of the putative inducer-binding site in OpuAR with respect to the affinity of choline and glycine betaine binding. Collectively, our data characterize GbsR-/OpuAR-type proteins as an extended sub-group within the MarR-superfamily of transcriptional regulators and identify a novel type of substrate-inducible import system for osmostress protectants.

Pod filling stage considers as a receiver (sink), which reflects plant performance during previous growth stages. In order to study, the influence of drought imposed at different growth stages, and the impact of foliar applied glycine betaine (GB) and proline on the status of osmolytes and antioxidant defense system of pea plant during pod filling stage, a field experiment was conducted in 2012/2013 and 2013/2014 on clay loam soil. Four different irrigation regimes were applied to provide drought at different growth stages: (1) vegetative stage, (2) flowering stage, as short-term drought stress, (3) throughout the stages of vegetative + flowering growth (long-term drought stress), and (4) control (without stress). Foliar applications of GB and proline at 4 mM for each, in addition to distilled water as control, were conducted. Generally, drought applications reduced the growth and yield of pea plant. Long-term drought was more effective to reduce growth and yield than drought at flowering stage. GB increased the yield and its soluble protein concentration more than proline. Proline recorded the maximum increase in non-enzymatic antioxidant defense system under drought. Application of GB or proline enhanced the activity of SOD, APX and catalase in leaves under drought, while in seeds they increased SOD activity under long-term drought stress. APX activity in seeds under drought decreased by GB application. The maximum positive effect was for GB under unstressed condition and drought at vegetative stage, by maximizing APX activity, in addition to enhancing the production and translocation of assimilates from source to sink.

The effects of salt and water stress on growth and several stress markers were investigated in cherry tomato plants. Some growth parameters (stem length and number of leaves) and chlorophyll contents were determined every third day during plant growth, and leaf material was collected after 25 and 33 days of treatment. Both stresses inhibited plant growth; chlorophyll levels, however, decreased only in response to high NaCl concentrations. Proline contents largely increased in leaves of stressed plants, reaching levels high enough to play a major role in cellular osmotic adjustment. Despite reports indicating that tomato does not synthesize glycine betaine, the stress-induced accumulation of this osmolyte was detected in cherry tomato, albeit at lower concentration than that of proline. Therefore, it appears that the plants are able to synthesise glycine betaine as a secondary osmolyte under strong stress conditions. Total sugars levels, on the contrary, decreased in stress-treated plants. Both stress treatments caused secondary oxidative stress in the plants, as indicated by a significant increase in malondialdehyde (MDA) contents. Water stress led to an increase in total phenolics and flavonoid contents and a reduction of carotenoid levels in the leaves; flavonoids also increased under high salinity conditions.

[EN] Gypsum areas are stressful environments inhabited by gypsophytes, plants that are exclusive for such habitats, and by plants that grow on gypsum but also on other soil types, the so-called gypsovags. To investigate possible differences between gypsovags and gypsophytes with respect to basic stress response mechanisms, two common osmolytes, glycine betaine and total soluble sugars, as well as monovalent (Na+ and K+) and bivalent (Ca2+ and Mg2+) cations, were quantified, under field conditions, in two Iberian endemic gypsophytes (Gypsophila struthium subsp. hispanica and Ononis tridentata) and two common Mediterranean gypsovags (Rosmarinus officinalis and Helianthemum syriacum). Their spatial variation according to a topographic gradient and their temporal variation over a period of three successive seasons were correlated with climatic data and soil characteristics. This analysis confirmed that water stress is the main environmental stress factor in gypsum habitats, whereas the percentage of gypsum in the soil does not seem to play any relevant role in the activation of stress responses in plants. Glycine betaine may contribute to stress tolerance in the gypsophytes, but not in the gypsovags, according to the close correlation found between the level of this osmolyte and the gypsophily of the investigated taxa. Cation contents in the plants did not correlate with those present in the soil, but the gypsophytes have higher levels of Ca2+ and Mg2+ than the gypsovags, under all environmental conditions, which may represent an adaptation mechanism to their specific habitat, This work has been supported by the Spanish Ministry of Science and Innovation (Project CGL2008-00438/BOS), with contribution from the European Regional Development Fund.

Plant responses to salinity are complex, especially when combined with other stresses, and involve many changes in gene expression and metabolic fluxes. Until now, plant stress studies have been mainly dealt only with a single stress approach. However, plants exposed to multiple stresses at the same time, a combinatorial approach reflecting real-world scenarios, show tailored responses completely different from the response to the individual stresses, due to the stress-related plasticity of plant genome and to specific metabolic modifications. In this view, recently it has been found that γ-aminobutyric acid (GABA) but not glycine betaine (GB) is accumulated in durum wheat plants under salinity only when it is combined with high nitrate and high light. In these conditions, plants show lower reactive oxygen species levels and higher photosynthetic efficiency than plants under salinity at low light. This is certainly relevant because the most of drought or salinity studies performed on cereal seedlings have been done in growth chambers under controlled culture conditions and artificial lighting set at low light. However, it is very difficult to interpret these data. To unravel the reason of GABA accumulation and its possible mode of action, in this review, all possible roles for GABA shunt under stress are considered, and an additional mechanism of action triggered by salinity and high light suggested. Plant responses to salinity are complex, especially when combined with other stresses, and involve many changes in gene expression and metabolic fluxes. Until now, plant stress studies have been mainly dealt only with a single stress approach. However, plants exposed to multiple stresses at the same time, a combinatorial approach reflecting real-world scenarios, show tailored responses completely different from the response to the individual stresses, due to the stress-related plasticity of plant genome and to specific metabolic modifications. In this view, recently it has been found that gamma-aminobutyric acid (GABA) but not glycine betaine (GB) is accumulated in durum wheat plants under salinity only when it is combined with high nitrate and high light. In these conditions, plants show lower reactive oxygen species levels and higher photosynthetic efficiency than plants under salinity at low light. This is certainly relevant because the most of drought or salinity studies performed on cereal seedlings have been done in growth chambers under controlled culture conditions and artificial lighting set at low light. However, it is very difficult to interpret these data. To unravel the reason of GABA accumulation and its possible mode of action, in this review, all possible roles for GABA shunt under stress are considered, and an additional mechanism of action triggered by salinity and high light suggested.

Members of the genus Spiribacter are found worldwide and are abundant in ecosystems possessing intermediate salinities between seawater and saturated salt concentrations. Spiribacter salinus M19-40 is the type species of this genus and its first cultivated representative. In the habitats of S. salinus M19-40, high salinity is a key determinant for growth and we therefore focused on the cellular adjustment strategy to this persistent environmental challenge. We coupled these experimental studies to the in silico mining of the genome sequence of this moderate halophile with respect to systems allowing this bacterium to control its potassium and sodium pools, and its ability to import and synthesize compatible solutes. S. salinus M19-40 produces enhanced levels of the compatible solute ectoine, both under optimal and growth-challenging salt concentrations, but the genes encoding the corresponding biosynthetic enzymes are not organized in a canonical ectABC operon. Instead, they are scrambled (ectAC; ectB) and are physically separated from each other on the S. salinus M19-40 genome. Genomes of many phylogenetically related bacteria also exhibit a non-canonical organization of the ect genes. S. salinus M19-40 also synthesizes trehalose, but this compatible solute seems to make only a minor contribution to the cytoplasmic solute pool under osmotic stress conditions. However, its cellular levels increase substantially in stationary phase cells grown under optimal salt concentrations. In silico genome mining revealed that S. salinus M19-40 possesses different types of uptake systems for compatible solutes. Among the set of compatible solutes tested in an osmostress protection growth assay, glycine betaine and arsenobetaine were the most effective. Transport studies with radiolabeled glycine betaine showed that S. salinus M19-40 increases the pool size of this osmolyte in a fashion that is sensitively tied to the prevalent salinity of the growth medium. It was amassed in salt-stressed cells in unmodified form and suppressed the synthesis of ectoine. In conclusion, the data presented here allow us to derive a genome-scale picture of the cellular adjustment strategy of a species that represents an environmentally abundant group of ecophysiologically important halophilic microorganisms. España Ministerio de Economía y Competitividad CGL2013-46941-P and CGL2017-83385-p

Durum wheat is one of the oldest and most important edible cereal crops and its cultivation has considerable economic importance in many countries. However, adverse conditions, such as high irradiance and increasing salinity of soils, could lead to a decrease in productivity over the next few decades. Durum wheat plants under salinityare able toaccumulate glycine betaine to osmotically balance the cytosol and reduce oxidative stress, especially in young tissues. However, the synthesis of this fundamental osmolyte is inhibited by high light in T. durum even under salinity. Choline monooxygenase is the first enzyme involved in the glycine betaine biosynthetic pathway. Thus, to explain the glycine betaine inhibition, we analyzed the effect of both salinity and high light on the putative TdCMO gene expression. Thirty-eight TdCMO different transcripts were isolated in the young leaves of durum wheat grown in different stress conditions. All translated amino acid sequences, except for the TdCMO1a6 clone, showed a frame shift caused by insertions or deletions. The presence of different transcripts could depend on the presence of duplicated genes, different allelic forms, and alternative splicing events. TdCMO1a6 computational modeling of the 3D structure showed that in durum wheat, a putative CMO-like enzyme with a different Rieske type motif, is present and could be responsible for the glycine betaine synthesis.

Cyclic di-adenosine monophosphate (c-di-AMP) is a recently discovered signaling molecule important for the survival of Firmicutes, a large bacterial group that includes notable pathogens such asStaphylococcus aureus. However, the exact role of this molecule has not been identified.dacA, theS. aureusgene encoding the diadenylate cyclase enzyme required for c-di-AMP production, cannot be deleted when bacterial cells are grown in rich medium, indicating that c-di-AMP is required for growth in this condition. Here, we report that anS. aureus dacAmutant can be generated in chemically defined medium. Consistent with previous findings, this mutant had a severe growth defect when cultured in rich medium. Using this growth defect in rich medium, we selected for suppressor strains with improved growth to identify c-di-AMP-requiring pathways. Mutations bypassing the essentiality ofdacAwere identified inalsTandopuD, encoding a predicted amino acid and osmolyte transporter, the latter of which we show here to be the main glycine betaine-uptake system inS. aureus. Inactivation of these transporters likely prevents the excessive osmolyte and amino acid accumulation in the cell, providing further evidence for a key role of c-di-AMP in osmotic regulation. Suppressor mutations were also obtained inhepS, hemB, ctaAandqoxB, coding for proteins required for respiration. Furthermore, we show thatdacAis dispensable for growth in anaerobic conditions. Together, these finding reveal an essential role for the c-di-AMP signaling network in aerobic, but not anaerobic, respiration inS. aureus.

We here characterized the stress-tolerant alfalfa microsymbiont Sinorhizobium meliloti B401. B401-treated plants showed high nitrogen fixation rates under humid and semiarid environments. The production of glycine betaine in isolated bacteroids positively correlated with low precipitation levels, suggesting that this compound acts as a critical osmoprotectant under field conditions. Genome analysis revealed that strain B401 contains alternative pathways for the biosynthesis and uptake of glycine betaine and its precursors. Such genomic information will offer substantial insight into the environmental physiology of this biotechnologically valuable nitrogen-fixingbacterium. Fil: Jozefkowicz, Cintia. Instituto Nacional de Tecnología Agropecuaria. Centro de Investigación en Ciencias Veterinarias y Agronómicas. Instituto de Genética; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina Fil: Brambilla, Silvina Maricel. Instituto Nacional de Tecnología Agropecuaria. Centro de Investigación en Ciencias Veterinarias y Agronómicas. Instituto de Genética; Argentina Fil: Frare, Romina Alejandra. Instituto Nacional de Tecnología Agropecuaria. Centro de Investigación en Ciencias Veterinarias y Agronómicas. Instituto de Genética; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina Fil: Stritzler, Margarita. Instituto Nacional de Tecnología Agropecuaria. Centro de Investigación en Ciencias Veterinarias y Agronómicas. Instituto de Genética; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina Fil: Piccinetti, Carlos. Instituto Nacional de Tecnología Agropecuaria. Centro de Investigación en Ciencias Veterinarias y Agronómicas. Instituto de Microbiología y Zoología Agrícola; Argentina Fil: Puente, Mariana L. Instituto Nacional de Tecnología Agropecuaria. Centro de Investigación en Ciencias Veterinarias y Agronómicas. Instituto de Microbiología y Zoología Agrícola; Argentina Fil: Berini, Carolina Andrea. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay. Instituto de Investigaciones Biomédicas en Retrovirus y Sida. Universidad de Buenos Aires. Facultad de Medicina. Instituto de Investigaciones Biomédicas en Retrovirus y Sida; Argentina Fil: Pérez, Pedro Reyes. Instituto Nacional de Tecnología Agropecuaria. Centro de Investigación en Ciencias Veterinarias y Agronómicas. Instituto de Genética; Argentina Fil: Soto, Gabriela Cynthia. Instituto Nacional de Tecnología Agropecuaria. Centro de Investigación en Ciencias Veterinarias y Agronómicas. Instituto de Genética; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina Fil: Ayub, Nicolás Daniel. Instituto Nacional de Tecnología Agropecuaria. Centro de Investigación en Ciencias Veterinarias y Agronómicas. Instituto de Genética; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina

This study was conducted to evaluate enzymatic and non-enzymatic antioxidant response of ‘Selva’ strawberry plants on exogenous nitric oxide under saline conditions with respect to time of application. Sodium nitroprusside (SNP), as nitric oxide (NO) source, was applied on the leaves by spray before, simultaneously, or after the initiation of saline stress. Results indicated that salinity and/or SNP at concentrations of 50 and 75 μM caused increase in activity of antioxidant enzymes, such as catalase, superoxide dismutase, glutathione reductase, ascorbate peroxidase and peroxidases as well as leaf content of proline, glycine betaine and total phenolics in comparison to control. Time of NO application was important because the highest levels of catalase and ascorbic peroxidase were in plants pre-treated with SNP one week before the initiation of salinity stress. Plants from these combinations had the highest fruit yield among all saline stressed plants. So, it seems that earlier application of SNP is more effective for an optimised protection against deleterious influence of salinity stress, because pre-treated plants had a sufficient time to develop an appropriate antioxidant response. The application of SNP simultaneously or after exposure of plants to stress conditions, was also helpful in increasing plant tolerance but to a lesser extent.

Barley (Hordeum vulgare L.) plants accumulate glycine betaine (GB), a major compatible solute, in response to salt stress. In barley, GB is produced by a two-step oxidation of choline in a cooperative way in the cytosol and peroxisomes. In this study, we investigated the localization of two GB biosynthetic enzymes, choline monooxygenase (CMO) and betaine aldehyde dehydrogenase (BADH), in the tissues of barley plants (cv. Haruna-nijyo) grown under normal and saline conditions. Three-week-old barley plants grown hydroponically were treated with a hydroponic culture solution containing 200 mM NaCl for 72 h. Salt treatment resulted in increased expression of CMO and BADH proteins mainly in the leaves of barley but not in the roots. The expression of CMO protein was increased by the presence of NaCl in younger leaves but decreased in older leaves. The tissue localization of CMO and BADH proteins was analyzed by immunofluorescent labeling method using their primary antibodies and a fluorescein-conjugated secondary antibody. CMO and BADH proteins were constitutively co-localized in mesophyll and bundle sheath cells under both normal and saline conditions. A possible physiological function of GB in the salt tolerance of barley plants is discussed.

Topoisomerases are unique enzymes having an ability to remove or add DNA supercoils and untangle the snarled DNA. They can cut, shuffle, and religate DNA strands and remove the torsional stress during DNA replication, transcription or recombination events. In the present study, we over-expressed topoisomerase II (TopoII) in tobacco (Nicotiana tabaccum) and examined its role in growth and development as well as salt (NaCl) stress tolerance. Several putative transgenic plants were generated and the transgene integration and expression was confirmed by PCR and Southern blot analyses, and RT-PCR analysis respectively. Percent seed germination, shoot growth, and chlorophyll content revealed that transgenic lines over-expressing the NtTopoIIα-1 gene exhibited enhanced tolerance to salt (150 and 200 mM NaCl) stress. Moreover, over-expression of TopoII lead to the elevation in proline and glycine betaine levels in response to both concentrations of NaCl as compared to wild-type. In response to NaCl stress, TopoII over-expressing lines showed reduced lipid peroxidation derived malondialdehyde (MDA) generation. These results suggest that TopoII plays a pivotal role in salt stress tolerance in plants.

The present experiment was aimed at examining the ameliorative effect of foliar-applied glycine betaine (50 mM GB) and glycine betaine containing sugarbeet extract (50 mM GB) on various physiological and biochemical attributes of okra plants under salt stress. The experiment comprised of two okra cultivars (Arka-anamika and Sabaz-pari), two salt levels (0 and 150 mM NaCl), and two GB sources (synthetic pure GB and sugarbeet extract) arranged in four replicates. Salt stress significantly suppressed the biomass production, yield, and different gas exchange attributes (A, E, Ci, and gs). Glycine betaine and proline contents in leaves, and Na+ and Cl− contents in both leaves and roots increased, while K+ and Ca2 + contents and K+/Na+ ratios decreased significantly. Foliar application of both pure GB and sugarbeet extract significantly reduced the adverse effects of salt stress on plant biomass production, plant yield, various gas exchange characteristics and leaf K+, Ca2 +, Cl− and Na+ contents. However, GB and sugarbeet extract showed differential effects on A, gs, E, Ci, Ci/Ca ratio, leaf K+, Ca2 +, and Cl− contents, and K+/Na+ ratio. Pure GB proved better than the sugarbeet extract in improving growth, while the reverse was true for plant yield under salt stress. However, with respect to different gas exchange attributes both GB and sugarbeet extract were found to be equally effective in reducing the adverse effects of salt stress on these photosynthetic attributes. Foliar-applied sugarbeet extract was found to be more effective as compared to pure GB in reducing the adverse effects of salt stress on K+ and Ca2 + uptake and K+/Na+ ratio in shoot and root of both okra cultivars. Thus, sugarbeet extract could be used to induce salt tolerance in economically important crop plants.

International audience; Among all of the phytomicronutrients, lipotropes which limit excess hepatic fat deposits, have been very little studied. And yet, liver steatosis is common to several chronic diseases. Among the lipotropes, betaine, choline, myo-inositol, methionine, magnesium, niacin, pantothenic acid and folates were the ones for which sufficient data have possibly been found to allow the selection of a significant number of plant-based foods (PBFs). Our objectives were to unravel the differences or similarities in the lipotrope density (LD) profile of raw PBFs and to define an index reflecting lipotrope contents. From databases for betaine and choline contents, we selected 56 raw PBFs (38 when inositol content was taken into consideration). Lipotropic capacity (LC) was defined as the means of the 8 LD profiles, each expressed as a percentage of raw asparagus LD, which has the highest mean ranking for the 8 LDs (LC = 100). LCs ranged from 7 (grapes) to 672% (spinach), relative to asparagus LC. Among cereal, fruit, legume and seed groups, quinoa, blackberry, common bean and sesame seed had the highest levels of LC (155%, 107%, 36% and 26%, respectively). On a 100 kcal-basis, vegetables are the best sources of lipotropes, followed by cereals, fruits and legumes, then nuts and seeds. PBF LD profiles were complementary but more diversified compared to animal-based food LD profiles. (C) 2011 Elsevier Inc. All rights reserved.

Submitted by Guilherme Lemeszenski (guilherme@nead.unesp.br) on 2013-08-22T18:47:05Z No. of bitstreams: 1 S0100-204X2009000900006.pdf: 1214882 bytes, checksum: fac04ec1488bb324f68f43410b23f9b4 (MD5) Made available in DSpace on 2013-08-22T18:47:05Z (GMT). No. of bitstreams: 1 S0100-204X2009000900006.pdf: 1214882 bytes, checksum: fac04ec1488bb324f68f43410b23f9b4 (MD5) Previous issue date: 2009-09-01 Made available in DSpace on 2013-09-30T17:49:58Z (GMT). No. of bitstreams: 2 S0100-204X2009000900006.pdf: 1214882 bytes, checksum: fac04ec1488bb324f68f43410b23f9b4 (MD5) S0100-204X2009000900006.pdf.txt: 31926 bytes, checksum: 261589ac7f82fc996572eee1c0133945 (MD5) Previous issue date: 2009-09-01 Submitted by Vitor Silverio Rodrigues (vitorsrodrigues@reitoria.unesp.br) on 2014-05-20T13:13:22Z No. of bitstreams: 2 S0100-204X2009000900006.pdf: 1214882 bytes, checksum: fac04ec1488bb324f68f43410b23f9b4 (MD5) S0100-204X2009000900006.pdf.txt: 31926 bytes, checksum: 261589ac7f82fc996572eee1c0133945 (MD5) Made available in DSpace on 2014-05-20T13:13:22Z (GMT). No. of bitstreams: 2 S0100-204X2009000900006.pdf: 1214882 bytes, checksum: fac04ec1488bb324f68f43410b23f9b4 (MD5) S0100-204X2009000900006.pdf.txt: 31926 bytes, checksum: 261589ac7f82fc996572eee1c0133945 (MD5) Previous issue date: 2009-09-01 O objetivo deste trabalho foi avaliar os indicadores fisiológicos da interação entre deficit hídrico e acidez do solo em plantas jovens de cana-de-açúcar. As plantas foram submetidas a três tratamentos de disponibilidade hídrica, medidos em percentagem de capacidade de campo (CC) - sem estresse (70% CC), estresse moderado (55% CC) e estresse severo (40% CC); e três tratamentos de acidez no solo, medidos em termos de saturação por bases (V) - baixa acidez (V = 55%), média acidez (V = 33%) e alta acidez (V = 23%). O experimento foi realizado em casa de vegetação a 29,7±4,3ºC e 75±10% UR. O delineamento experimental utilizado foi o de blocos ao acaso, em esquema fatorial 3x3, com quatro repetições. Após 60 dias, foram determinados os teores de solutos compatíveis - trealose, glicina betaína e prolina - na folha diagnóstico e o crescimento inicial da parte aérea. Os solutos compatíveis trealose, glicina betaína e prolina são indicadores do efeito da interação dos estresses hídrico e ácido no solo. O acúmulo dos solutos compatíveis nos tecidos foliares das plantas não é capaz de impedir a redução na produção de matéria seca da cana-de-açúcar, resultante do agravamento nas condições de disponibilidade hídrica e de acidez no solo. The aim of this work was to assess the physiological indicators of the interaction between water deficit and soil acidity, in sugarcane. The plants were submitted to three treatments of water availability - no stress (70% of field capacity, FC), moderate stress (55% FC), and extreme stress (40% FC); and three acidity treatments - no acidity [base saturation (V) = 55%], average acidity (V = 33%), and high acidity (V = 23%). The experiment was carried out in greenhouse, with 29.7±4.3ºC and 75±10% RH. The experimental design was in randomized blocks, in 3x3 factorial arrangement, with four replicates. After 60 days, the contents of compatible solutes - trehalose, glycine betaine and proline - in the diagnostic leaf and the initial growth of shoots were determined. The compatible solutes trehalose, glycine betaine, and proline are indicators of the interaction of water and acidity stresses in the soil. The accumulation of compatible solutes in plant foliar tissues can not prevent sugarcane losses in dry matter production, caused by increasing water deficit and soil acidity. Agência Paulista de Tecnologia dos Agronegócios Universidade Estadual Paulista Faculdade de Ciências Agrárias e Veterinárias Departamento de Biologia Aplicada à Agropecuária Universidade Estadual Paulista Faculdade de Ciências Agrárias e Veterinárias Departamento de Biologia Aplicada à Agropecuária

We have previously shown that the C-terminal cystathionine beta-synthase (CBS) domains of the nucleotide-binding domains of the ABC transporter OpuA, in conjunction with an anionic membrane surface function, act as sensor of internal ionic strength (I(in)). Here, we show that a surface-exposed cationic region in the CBS module domain is critical for ion sensing. The consecutive substitution of up to five cationic residues led to a gradual decrease of the ionic strength dependence of transport. In fact, a 5-fold mutant was essentially independent of salt in the range from 0 to 250 mM KCl (or NaCl), supplemented to medium of 30 mM potassium phosphate. Importantly, the threshold temperature for transport was lowered by 5-7 degrees C and the temperature coefficient Q(10) was lowered from 8 to similar to 1.5 in the 5-fold mutant, indicating that large conformational changes are accompanying the CBS-mediated regulation of transport. Furthermore, by replacing the anionic C-terminal tail residues that extend the CBS module with histidines, the transport of OpuA became pH-dependent, presumably by additional charge interactions of the histidine residues with the membrane. The pH dependence was not observed at high ionic strength. Altogether the analyses of the CBS mutants support the notion that the osmotic regulation of OpuA involves a simple biophysical switching mechanism, in which nonspecific electrostatic interactions of a protein module with the membrane are sufficient to lock the transporter in the inactive state.

Aims: To investigate the effect of glycine betaine (GB) on the survival of Listeria monocytogenes on leaf surfaces under low relative humidity (RH). Methods and Results: The addition of GB (≥25 mmol l−1) improved the survival of L. monocytogenes under low RH on parsley leaves, thus suggesting that GB can improve the tolerance of L. monocytogenes to desiccation. Ten times less GB was needed to improve L. monocytogenes survival under low RH on nonbiological surfaces compared with parsley leaves, suggesting that, on the leaf surface, L. monocytogenes may have to compete for the available GB with autochthonous bacteria and/or the plant itself. Wild type and mutants carrying deletions in the three GB uptake systems, BetL, Gbu and OpuC, behaved similarly with and without added GB on parsley leaves (P > 0·05). In addition, preaccumulation of GB, triggered by osmotic stress prior to inoculation, failed to improve survival under low RH compared with osmotic stress without GB accumulation. Conclusions: Exogenous GB had a protective effect on L. monocytogenes cells from desiccation during survival on parsley leaves. This effect was independent of intracellular GB accumulation by the known uptake systems. Significance and Impact of the Study: Presence of GB could improve the survival of L. monocytogenes to desiccation on leaf surfaces and nonbiological surfaces.

Zinc (Zn) pollution in the soil represents a major problem for crop production worldwide. In the present work, two horticultural plants exhibiting different tolerance to Zn, Lactuca sativa cv. Phillipus and Brassica oleracea cv. Bronco, were exposed to Zn to evaluate the contribution of compatible osmolytes such as proline (Pro), glycine betaine (GB) and c-aminobutyric acid (GABA) in the mechanism(s) of tolerance to Zn stress. This study confirms the higher susceptibility of L. sativa to Zn stress: lettuce plants experienced a strong reduction in biomass, while the levels of Pro and GB increased. These results suggest that in L. sativa, the increase of Pro and GB does not represent a mechanism of resistance to toxicity, but it is likely a symptom of Zn stress. Conversely, in B. oleracea, a slight decrease in Pro levels, mainly catalysed by degradation through proline dehydrogenase, was observed; a similar behaviour affected GB levels. On the other hand, GABA synthesis was slightly, but significantly, increased. The presence of high levels of GABA in Zn-stressed B. oleracea would suggest that reactive oxygen species detoxification could be essential to improve the resistance to toxicity under metal stress conditions.