Ameliorative effects of potassium on drought-induced decreases in fiber length of cotton(Gossypium hirsutum L.) are associated with osmolyte dynamics during fiber development

Fiber length of cotton(Gossypium hirsutum L.)decreases under drought stress,potassium(K)could diminish the decreased caused by drought,but the mechanism associated with this alleviation effect is not clear.We evaluated the effect of K on fiber elongation using two cotton cultivars,Simian 3 and Siza 3,grown in well-watered and drought-stressed conditions.Potassium fertilizer(K2O)was applied 0,150,or 300 kg ha?1 in each growing condition.Drought stress reduced the final fiber length due to a decline in the maximum rate of rapid elongation(Vmax,mmday?1).The application of K alleviated the droughtinduced fiber length reduction by increasing Vmax.At 10 and 15 days post-anthesis(DPA),drought significantly reduced osmotic potential(OP)and increased K+and malate contents at all K rates,relative to well-watered conditions,which was associated with increased activities of phosphoenolpyruvate carboxylase(PEPC),V-ATPase,PPase,and PM H+-ATPase in cotton fiber.However,the relative contribution of K+and malate to OP declined under drought in comparison with well-watered condition.Compared with control without K,K application decreased OP and increased the accumulation of osmolytes(K+,malate and soluble sugar)as well as the activities of related enzymes in fiber irrespective of water treatments.Moreover,K application increased osmotic adjustment during drought,and improved the contribution of K+and malate to OP,especially under drought stress.This study showed that drought decreased fiber length by reducing Vmax,and K application ameliorates the decline in fiber elongation due to drought by enhancing osmolytes accumulation and their contribution to OP in fiber cells.

The osmolyte-producing endophyte Streptomyces albidoflavus OsiLf-2 induces drought and salt tolerance in rice via a multi-level mechanism

Drought and salinity are major environmental stresses that impair crop growth and productivity worldwide. Improving drought and salt tolerance of crops with microbial mutualists is an effective and environmentally sound strategy to meet the demands of the ever-growing world population. In the present study, we found that the Streptomyces albidoflavus Osi Lf-2, a moderately salt-tolerant endophytic actinomycete, produced abundant osmolytes, including proline, polysaccharides, and ectoine. Inoculation with Osi Lf-2 increased the osmotic-adjustment ability of the rice host by increasing the proline content(by250.3% and 49.4%) and soluble sugar(by 20.9% and 49.4%) in rice under drought and salt conditions, relative to the uninoculated control. Osi Lf-2 increased stress responses in the rice host at the physiological and biochemical levels(photosynthesis efficiency, osmolytes and antioxidant content), and the gene level(osmolytes synthesis, stress-responsive and ion-transport related genes), raising rice yields under both greenhouse and saline–alkaline soil conditions. The use of endophytic actinomycetes offers a promising biotechnological approach to developing stress-tolerant plants.

TaNF-YB11,a gene of NF-Y transcription factor family in Triticum aestivum,confers drought tolerance on plants via modulating osmolyte accumulation and reactive oxygen species homeostasis

Transcription factors(TFs)regulate diverse stress defensive-associated physiological processes and plant stress responses.We characterized TaNF-YB11,a gene of the NF-YB TF family in Triticum aestivum,in mediating plant drought tolerance.TaNF-YB11 harbors the conserved domains specified by its NF-YB partners and targets the nucleus after the endoplasmic reticulum(ER)assortment.Yeast two-hybrid assay indicated the interactions of TaNF-YB11 with TaNF-YA2 and TaNF-YC3,two proteins encoded by genes in the NF-YA and NF-YC families,respectively.These results suggested that the heterotrimer established among them further regulated downstream genes at the transcriptional level.The transcripts of TaNF-YB11 were promoted in roots and leaves under a 27-h drought regime.Moreover,its upregulated expression levels under drought were gradually restored following a recovery treatment,suggesting its involvement in plant drought response.TaNF-YB11 conferred improved drought tolerance on plants;the lines overexpressing target gene displayed improved phenotype and biomass compared with wild type(WT)under drought treatments due to enhancement of stomata closing,osmolyte accumulation,and cellular reactive oxygen species(ROS)homeostasis.Knockdown expression of TaP5CS2,a P5CS family gene modulating proline biosynthesis that showed upregulated expression in drought-challenged TaNF-YB11 lines,alleviated proline accumulation of plants treated by drought.Likewise,TaSOD2 and TaCAT3,two genes encoding superoxide dismutase(SOD)and catalase(CAT)that were upregulated underlying TaNF-YB11 regulation,played critical roles in ROS homeostasis via regulating SOD and CAT activities.RNA-seq analysis revealed that numerous genes associated with processes of‘cellular processes',‘environmental information processing',‘genetic information processing',‘metabolism',and‘organismal systems'modified transcription under drought underlying control of TaNF-YB11.These results suggested that the TaNF-YB11-mediated drought response is possibly accomplished through the target gene in modifying gene transcription at the global level,which modulates complicated biological processes related to drought response.TaNF-YB11 is essential in plant drought adaptation and a valuable target for molecular breeding of drought-tolerant cultivars in T.aestivum.

Osmolyte modulated enhanced rice leaf catalase activity under salt-stress

Change in catalase activity was examined in leaves of rice plant exposed to salinity. Depending on the method of preparation of crude protein extract from leaf and the constituents of the assay medium, a significant difference in enzyme activity was recorded. Inclusion of sorbitol or mannitol or sucrose in the extraction and enzyme assay medium enhanced the enzyme activity in salt-stressed samples by nearly 1.5-1.8 fold, compared to the activity found in un- stressed plants, which otherwise showed a 50% declined activity in leaf extract prepared in buffer solution and assayed in a medium depleted of these sugars. In view of the accumulation of osmolytes under saline condition, these observations suggest that the catalase activity is modulated by the osmolytes and maintains a high rate of hydrogen peroxide scavenging property in vivo and serves as the major antioxidant enzyme to scavenge the salt-induced formation of peroxide. Therefore, the salt-stress induced appearance of low activity of the enzyme under normal buffer extraction and assay conditions, as reported in literature may represent an apparent than for its real in vivo activity.

Accumulation of Polyphenolic Compounds and Osmolytes under Dehydration Stress and Their Implication in Redox Regulation in Four Indigenous Aromatic Rice Cultivars

Present work was undertaken to screen some drought tolerant indigenous aromatic rice cultivars(IARCs),commonly cultivated in West Bengal,India,based on their capacity to produce osmolytes,redox-sensitive phenolic acids and flavonoids,as contrivances for redox-regulation under drought stress.Polyethylene glycol induced post imbibitional dehydration stress mediated changes in redox regulatory properties of the germinating seeds of the four IARCs(Jamainadu,Tulaipanji,Sitabhog,Badshabhog),which were assessed in terms of changes in prooxidant accumulation(in-situ localization of reactive oxygen species(ROS)by confocal microscopy,DCFDA(2′,7′-dichlorofluorescin diacetate)oxidation,O2-and H2O2 accumulation),cumulative antioxidative defense(radical scavenging property and total thiol content),ROS scavenging phenolic acids(gallic acid,protocatechuic acid,gentisic acid,para-hydroxy benzoic acid,chlorogenic acid,caffeic acid,syringic acid,salicylic acid,sinapic acid and p-coumaric acid)and flavonoids(catechin,naringin,rutin,quercetin,kaempferol,myricetin and apigenin).The capability of germinating seeds to accumulate osmolytes(like glycinebetaine,proline,soluble carbohydrates and K+ion)and polyphenolic compounds was also correlated with their corresponding redox status and redox biomarkers(conjugated diene,hydroperoxide,thiobarbituric acid reactive substances and free carbonyl content)produced under the same conditions.The results in general showed that accumulation of osmolytes along with the redox-sensitive phenolics and flavonoids conferred the ability to maintain the redox homeostasis under drought stress for the tolerant IARCs(Badshabhog and Tulaipanji).

Osmolyte transport in Staphylococcus aureus and the role in pathogenesis

Osmolyte transport is a pivotal part of bacterial life, particularly in high salt environments. Several low and high affinity osmolyte transport systems have been identified in various bacterial species. A lot of research has centered on characterizing the osmolyte transport systems of Gram-negative bacteria, but less has been done to characterize the same transport systems in Gram-positive bacteria. This review will focus on the previous work that has been done to understand the osmolyte transport systems in the species Staphylococcus aureus and how these transporters may serve dual functions in allowing the bacteria to survive and grow in a variety of environments, including on the surface or within humans or other animals.

Trehalose:A sugar molecule involved in temperature stress management in plants

Trehalose(Tre)is a non-reducing disaccharide found in many species,including bacteria,fungi,invertebrates,yeast,and even plants,where it acts as an osmoprotectant,energy source,or protein/membrane protector.Despite relatively small amounts in plants,Tre concentrations increase following exposure to abiotic stressors.Trehalose-6-phosphate,a precursor of Tre,has regulatory functions in sugar metabolism,crop production,and stress tolerance.Among the various abiotic stresses,temperature extremes(heat or cold stress)are anticipated to impact crop production worldwide due to ongoing climate changes.Applying small amounts of Tre can mitigate negative physiological,metabolic,and molecular responses triggered by temperature stress.Trehalose also interacts with other sugars,osmoprotectants,amino acids,and phytohormones to regulate metabolic reprogramming that underpins temperature stress adaptation.Transformed plants expressing Tre-synthesis genes accumulate Tre and show improved stress tolerance.Genome-wide studies of Tre-encoding genes suggest roles in plant growth,development,and stress tolerance.This review discusses the functions of Tre in mitigating temperature stress—highlighting genetic engineering approaches to modify Tre metabolism,crosstalk,and interactions with other molecules—and in-silico approaches for identifying novel Tre-encoding genes in diverse plant species.We consider how this knowledge can be used to develop temperature-resilient crops essential for sustainable agriculture.

Transcriptomic and physiological analyses identifying Lanzhou lily(Lilium davidii var.unicolor)drought adaptation strategies

Drought stress is the main limiting plant growth factor in arid and semiarid regions.The Lanzhou lily(Lilium davidii var.unicolor)is the only sweet-tasting lily grown in these regions of China that offers highly edible,medicinal,health,and ornamental value.The Tresor lily is an ornamental flower known for its strong resistance.Plants were grown under three different drought intensity treatments,namely,being watered at intervals of 5,15,and 25 d(either throughout the study or during specific growth stages).We measured the biomass,leaf area,photosynthetic response,chlorophyll content(SPAD value),and osmoregulation of both the Lanzhou lily and the Tresor lily(Lilium‘Tresor’).Additionally,we employed RNA sequencing(RNA-Seq)and qRT-PCR to investigate transcriptomic changes of the Lanzhou lily in response to drought stress.Results showed that under drought stress,the decreasing rate in the Lanzhou lily bulb weight was lower than the corresponding Tresor lily bulb rate;the net photosynthetic rate,transpiration rate,and stomatal conductance of the Lanzhou lily were all higher compared to the Tresor lily;osmoregulation constituents,such as glucose,fructose,sucrose,trehalose,and soluble sugar,in the Lanzhou lily were comparatively higher;PYL,NCED,and ERS genes were significantly expressed in the Lanzhou lily.Under moderate drought,the biosynthesis of flavonoids,circadian rhythms,and the tryptophan metabolism pathway of the Lanzhou lily were all significant.Under severe drought stress,fatty acid elongation,photosynthetic antenna protein,plant hormone signal transduction,flavone and flavonol biosynthesis,and the carotenoid biosynthesis pathway were all significant.The Lanzhou lily adapted to drought stress by coordinating its organs and the unique role of its bulb,regulating photosynthesis,increasing osmolyte content,activating circadian rhythms,signal transduction,fatty acid elongation metabolism,and phenylalanine and flavonoid metabolic pathways,which may collectively be the main adaptation strategy and mechanisms used by the Lanzhou lily under drought stress.

Using NMR-detected hydrogen-deuterium exchange to quantify protein stability in cosolutes,under crowded conditions in vitro and in cells

We review the use of nuclear magnetic resonance(NMR)spectroscopy to assess the exchange of amide protons for deuterons(HDX)in efforts to understand how high concentration of cosolutes,especially macromolecules,affect the equilibrium thermodynamics of protein stability.HDX NMR is the only method that can routinely provide such data at the level of individual amino acids.We begin by discussing the properties of the protein systems required to yield equilibrium thermodynamic data and then review publications using osmolytes,sugars,denaturants,synthetic polymers,proteins,cytoplasm and in cells.

植物盐诱导基因的研究进展简述

本文介绍了有关盐诱导基因包括跨膜通道蛋白基因、Ｏｓｍｏｌｙｔｅ生物合成基因等进展；随着功能基因组学的开展，利用基因表达的系统分析、ｃＤＮＡ微阵列、ＤＮＡ芯片、蛋白组技术等基于转座子标签和Ｔ－ＤＮＡ标签的反求遗传学技术等将为寻找及验证耐盐基因的功能提供技术保证。

溶剂特性对三臂星型类弹性蛋白多肽相变温度的影响

本文利用Spy Tag/Spy Catcher特性构建了三臂星型结构的类弹性蛋白多肽（elastin-like polypeptides,ELPs）,考察其在不同溶剂,如分子拥挤试剂、osmolytes及深共熔溶剂（deep eutectic solvents,DESs）中的相变温度及行为,并与含有相同ELPs重复数的线性ELPs120作对比。结果表明：在不同浓度拥挤试剂PEG2000作用下,两种结构的ELPs相变温度均降低,当其各自浓度均为25μmol/L时,三臂星型ELPs相变温度降低3℃-13℃,而ELPs120相变温度仅降低1.5℃-10.8℃。此外,在添加PEG2000后,三臂星型ELPs相变缓慢;在不同类型和浓度的osmolytes溶液中,25μmol/L三臂星型ELPs相变温度明显要比线性ELPs高8℃左右;在DESs体系中,三臂星型ELPs有类似与水溶液中的相变行为,且其相变温度受到抑制,另外三臂星型ELPs和ELPs120在DESs/PBS体系中,与在（氯化胆碱＋尿素）/PBS体系中的相变行为一致,其中当DESs体积含量为50%,ELPs120相变温度是最低的。由于ELPs在非单一缓冲液体系中的相变行为不同,这丰富了ELPs作为纯化标签的应用,且在非单一缓冲液体系中因降低了相变温度,节约了纯化融合蛋白的经济成本,同时也为研究ELPs拓扑结构与其相变行为之间的关系奠定理论基础。

Screening for Osmotic Stress Responses in Rice Varieties under Drought Condition

Drought is the major abiotic stress factor that limits rice production worldwide.To evaluate the osmotic stress responses in rice varieties under drought condition,a total of 42 high-yielding rice varieties were collected from various research stations of Kerala Agricultural University in India.The experimental setup comprises of initial hydroponic treatments at different osmotic potentials,artificially induced by desired strengths of polyethylene glycol(PEG6000),and followed by the pot planted experiments in the rain-out-zone.The activities of antioxidant enzymes,relative water content,cell membrane stability,photosynthetic pigments,proline content,along with plant growth parameters of the varieties under drought condition were evaluated.Moreover,the standard scores of these rice varieties were assessed under stress and recovery conditions based on the scoring scale of the Standard Evaluation System for rice.Among the 42 rice varieties,we identified 2 rice varieties,Swarnaprabha and Kattamodan,with less leaf rolling,better drought recovery ability as well as relative water content,increased membrane stability index,osmolyte accumulation,and antioxidant enzyme activities pointed towards their degree of tolerance to drought stress.The positive adaptive responses of these rice varieties towards drought stress can be used in the genetic improvement of rice drought resistance breeding program.

Water Stress Effects on Leaf Growth and Chlorophyll Content but Not the Grain Yield in Traditional Rice (<i>Oryza sativa</i>Linn.) Genotypes of Assam, India II. Protein and Proline Status in Seedlings under PEG Induced Water Stress

Abiotic stresses can directly or indirectly affect the physiological status of an organism by altering its metabolism, growth, and development. The leaf growth and Chlorophyll content has significantly shown to vary from the control ones while the grain yield was not affected. While many plant species naturally accumulate proline and protein as major organic osmolytes when subjected to different abiotic stresses. These compounds are thought to play adaptive roles in mediating osmotic adjustment and protecting sub cellular structures in stressed plants. Different approaches have been contemplated to increase the concentrations of proline like compounds in plants grown under stress conditions to increase their stress tolerance. Seven different traditional rice varieties of Assam were evaluated for their response to osmolyte production under physiological drought condition through simulation at three levels of osmotic stress of 0.15 bar, 0.25 bar and 0.56 bar of physiological drought initiated by polyethylene glycol (PEG 6000). Along with the evaluation for osmolyte response the different components of genotypic variation for six different drought-sustaining characters in the seven rice varieties were also substantiated. The results indicated that plant height and seed number have significant genotypic coefficient of variability (GCV) and heritability. Verities like Laodubi, Leserihali, Beriabhanga and Borah were screened out as the best drought sustaining variety.

Time course of physiological,biochemical,and gene expression changes under short-term salt stress in Brassica juncea L.

Salinity-imposed limitations on plant growth are manifested through osmotic and ionic imbalances. However, because salinity-induced responses vary considerably among crop plants, monitoring of such responses at an early stage has relevance. In this study,physiological(seed germination, seed vigor index, root length, shoot length, fresh weight,dry weight) and biochemical attributes(osmoprotectants, K^+/Na^+ ratio) were analyzed for a time-course assessment of salt responses in Indian mustard(Brassica juncea L.) with an emphasis on early monitoring. The results showed strong correlations for total soluble sugars at germination phase(24 h), proline content in the seedling establishment phase(48 h) and various physiological parameters including seed vigor index(R^2= 0.901), shoot length(R^2= 0.982), and fresh weight(R^2= 0.980) at 72 h(adaptation under stress). In addition, transcriptional changes were observed under NaCl treatment for key genes belonging to the family of selective ion transporters(NHX, HKT) and abscisic acid synthesis(AAO-3). The status of mitochondrial respiration was also examined as a probe for salinity tolerance at an early stage. The results suggested that although all the analyzed parameters showed correlations(negative or positive) with salt stress magnitude, their critical response times differed, with most of the studied biochemical, physiological, or molecular markers providing valuable information only after radicle emergence, whereas mitochondrial respiration via alternative oxidase was useful for the early detection of salt responses.

Targeting Glycinebetaine for Abiotic Stress Tolerance in Crop Plants:Physiological Mechanism,Molecular Interaction and Signaling

In the era of climate change,abiotic stresses(e.g.,salinity,drought,extreme temperature,flooding,metal/metalloid(s),UV radiation,ozone,etc.)are considered as one of the most complex environmental constraints that restricts crop production worldwide.Introduction of stress-tolerant crop cultivars is the most auspicious way of surviving this constraint,and to produce these types of tolerant crops.Several bioengineering mechanisms involved in stress signaling are being adopted in this regard.One example of this kind of manipulation is the osmotic adjustment.The quarternary ammonium compound glycinebetaine(GB),also originally referred to as betaine is a methylated glycine derivative.Among the betaines,GB is the most abundant one in plants,which is mostly produced in response to dehydration caused by different abiotic stresses like drought,salinity,and extreme temperature.Glycinebetaine helps in decreased accumulation and detoxification of ROS,thereby restoring photosynthesis and reducing oxidative stress.It takes part in stabilizing membranes and macromolecules.It is also involved in the stabilization and protection of photosynthetic components,such as ribulose-1,5-bisphosphate carboxylase/oxygenase,photosystem II and quarternary enzyme and protein complex structures under environmental stresses.Glycinebetaine was found to perform in chaperone-induced protein disaggregation.In addition,GB can confer stress tolerance in very low concentrations,and it acts in activating defense responsive genes with stress protection.Recently,field application of GB has also shown protective effects against environmental adversities increasing crop yield and quality.In this review,we will focus on the role of GB in conferring abiotic stress tolerance and the possible ways to engineer GB biosynthesis in plants.

Comparative Response of Red and Green Algae to the Quality of Coastal Water of Red Sea, Haql, Saudi Arabia

Aquatic plants are always exposed to various types of stresses and the marine algae have been considered as useful bioindicators for detecting different kinds of environmental alterations. Present investigation was carried out to test the comparative response of red algae (Gracilaria salicornia and Digenea simplex) and green algae (Enteromorpha compressa and Sargassum muticum) to the contaminants present in their surrounding growth medium. The results of the study show that the studied macroalgae responded differently in terms of physiological and biochemical parameters. Green algae exhibited higher concentration of Chl a, b, total chlorophyll content and Chl a:b ratio and carbohydrates content. Whereas, red algae showed higher content of carotenoids, phycocyanin and phycoerythrin and protein content. Moreover, red algae G. salicornia and D. simplex showed lower level of hydrogen peroxide content and TBARS and higher values of proline and glycine betaine content and activities of antioxidant enzymes viz. superoxide dismutase, peroxidase, and catalase than the green algae. Taking all these together, it can be concluded that red algae possess strong defense system than the green algae. Among the studied species, red algae G. salicornia was found best in having a stronger defense system.

Evaluation of subtropical ornamental trees for reclaiming salinity affected lands

This study was conducted to evaluate the tolerance of 1-year-old seedlings of ten subtropical ornamental tree species against a range of salinity levels of Na Cl from May 2015 to October 2015. The levels were further enhanced from November to April 2017 as 100% survival was observed in the initial concentrations for all species.The seedlings were grown during the first week of April2015 in 1000 earthen pots containing soil: farmyard manure(2:1), irrigated with tap water for 1 month followed by saline irrigation in May by maintaining electrical conductivity at 0.75, 1.00, 1.25, 1.50, 2.25, and 3.00 d S/m for 30,40, 50, 60, 90, and 120 m M Na Cl. Every 3 months,heights, relative leaf water content, and percent survival were determined;total soluble sugars of the upper leaves of each tree were quantified. All species exhibited consistent early growth and survival when supplied with 30, 40, 50 and 60 m M of Na Cl. Koelreutaria paniculata, Ficus benjamina, Putranjiva roxburghii, Bauhinia purpurea and Millettia ovalifolia were sensitive to elevated salinity levels and did not survive at the highest salt concentrations of 90 and 120 m M.

Clustering of halophytic species from Cyprus based on ionic contents

This paper presents the work conducted on the chemical constituents of some common and widely distributed halophyte taxa from Cyprus with the aim that these studies will help in the evaluation of halophytes for different economical purposes.The plant species of Crithmum maritimum L.,Limbarda crithmoides(L.)Dumort,Atriplex portulacoides L.,Salsola kali L.,Atriplex halimus L.,Limonium oleifolium Mill.,L.meyeri(Boiss.)Kuntze;and Tetraena alba(L.f.)Beier&Thulin were collected in the middle of July.The shoot tissue and leaf samples were collected from the natural habitats and left for drying under air circulation followed by placing them in oven at 60°C for 96 hours.The material was crushed using mortar and pestle and subjected to an analysis of macro-and micro-nutrients and biochemical compounds.K+/Na+in the leaf tissues of the dicot species showed relatively high values depicting their behavior as Na+includes but very low Cl-levels were recorded.Out of the species investigated here in 4 TFAA content was rather high.Values ranging from 0.5%to 1%dry weight were exhibited in one species.However,only 3 species showed very low TFAA values.Later may be due to low nitrogen availability in their environment.The phenetic analyses of eight halophyte species performed on the data matrix using Ntsys-pc program version 2.1 revealed that,cluster analysis of the overall results obtained here leads to 2 clusters.This discrimination appears to be as a result of their different abilities to accumulate either proline or glycine betaine.

Implication of Ions and Organic Solutes Accumulation in Amaranth (<i>Amaranthus cruentus</i>L.) Salinity Resistance

Salinity is one of the major environmental constraints limiting agricultural productivity in the world. The effects of salt stress on growth, ions and organic solutes accumulation were investigated in two amaranth (Amaranthus cruentus) cultivars: Rouge (salt-resistant) and Locale (salt-sensitive). Young plants of these cultivars were exposed, in hydroponic system, to three concentrations of NaCl: 0, 30 and 90 mM. Growth parameters, ions, free proline and soluble sugars concentrations were determined after 2 weeks of stress. NaCl effect resulted in plant growth reduction in both cultivars but plants of cultivar Rouge were less affected compared to that of cv. Locale. Na+, proline and soluble sugars concentrations increased significantly in leaves and roots under salinity while K+, Ca2+ and Mg2+ concentrations decreased in both cultivars. Proline and soluble sugars increased significantly in leaves and roots of cultivar Locale whereas in cultivar Rouge, proline increase was significant only in roots and soluble sugars increase was significant only in leaves. The highest increase of Na+ concentration occurred in leaves of cv. Rouge coupled with the lowest reduction in K+ concentration. The highest increase of proline occurred in leaves of cultivar Locale whereas the highest increase of soluble sugars was observed in leaves of cultivar Rouge. The reduction of the Ca2+ concentration under salt stress was more accentuated in both leaves and roots of cultivar Rouge than cultivar Locale while cv. Rouge maintained higher content in Mg2+ either in leaves or in roots in the presence of NaCl than cultivar Locale. These results suggest an implication of Na+, K+ and Mg2+ in salt resistance in these cultivars and that soluble sugars may play an important role in salt-resistance in Amaranthus cruentus. However, proline appears as a symptom of injury in stressed plants rather than an indicator of resistance.

Strategies to stabilize compact folding and minimize aggregation of antibody-based fragments

Monoclonal antibodies (mAbs) have proven to be useful for development of new therapeutic drugs and diagnostic techniques. To overcome the difficulties posed by their complex structure and folding, reduce undesired immunogenicity, and improve pharmacoki- netic properties, a plethora of different Ab fragments have been developed. These include recombinant Fab and Fv segments that can display improved properties over those of the original mAbs upon which they are based. Antibody (Ab) fragments such as Fabs, scFvs, diabodies, and nanobodies, all contain the variable Ig domains responsible for binding to specific antigenic epitopes, allowing for specific targeting of pathological cells and/or molecules. These fragments can be easier to produce, purify and refold than a full Ab, and due to their smaller size they can be well absorbed and distributed into target tissues. However, the physicochemical and structural properties of the immunoglobulin (Ig) domain, upon which the folding and conformation of all these Ab fragments is based, can limit the stability of Ab-based drugs. The Ig domain is fairly sensitive to unfolding and aggregation when produced out of the structural context of an intact Ab molecule. When unfolded, Ab fragments may lose their specificity as well as establish non-native interactions leading to protein aggregation. Aggregated antibody fragments display altered pharmacokinetic and immunogenic properties that can augment their toxicity. Therefore, much effort has been placed in understanding the factors impacting the stability of Ig folding at two different levels: 1) intrinsically, by studying the effects of the amino acid sequence on Ig folding;2) extrinsically, by determining the environmental conditions that may influence the stability of Ig folding. In this review we will describe the structure of the Ig domain, and the factors that impact its stability, to set the context for the different approaches currently used to achieve stable recombinant Ig domains when pursuing the development of Ab fragment-based biotechnologies.