Nitric oxide-mediated alleviation of arsenic stress involving metalloid detoxification and physiological responses in rice (Oryza sativa L.).

Rice is a staple crop, and food chain contamination of arsenic in rice grain possesses a serious health risk to billions of population. Arsenic stress negatively affects the rice growth, yield and quality of the grains. Nitric oxide (NO) is a major signaling molecule that may trigger various cellular responses in plants. The protective role of NO during arsenite (AsIII) stress and its relationship with plant physiological and metabolic responses is not explored in detail. Exogenous NO, supplemented through the roots in the form of sodium nitroprusside, has been shown to provide protection vis-à-vis AsIII toxicity. The NO-mediated variation in physiological traits such as stomatal density, size, chlorophyll content and photosynthetic rate maintained the growth of the rice plant during AsIII stress. Besides, NO exposure also enhanced the lignin content in the root, decreased total arsenic content and maintained the activities of antioxidant isoenzymes to reduce the ROS level essential for protecting from AsIII mediated oxidative damage in rice plants. Further, NO supplementation enhanced the GSH/GSSG ratio and PC/As molar ratio by modulating PC content to reduce arsenic toxicity. Further, NO-mediated modulation of the level of GA, IAA, SA, JA, amino acids and phenolic metabolites during AsIII stress appears to play a central role to cope up with AsIII toxicity. The study highlighted the role of NO in AsIII stress tolerance involving modulation of metalloid detoxification and physiological pathways in rice plants.

Root system architecture, physiological analysis and dynamic transcriptomics unravel the drought-responsive traits in rice genotypes.

Drought is the major abiotic factors that limit crop productivity worldwide. To withstand stress conditions, plants alter numerous mechanisms for adaption and tolerance. Therefore, in the present study, 106 rice varieties were screened for drought tolerance phenotype via exposing different concentrations of polyethylene glycol 6000 (PEG) in the hydroponic nutrient medium at the time interval of 1, 3, and 7 days to evaluate the changes in their root system architecture. Further, based on root phenotype obtained after PEG-induced drought, two contrasting varieties drought-tolerant Heena and -sensitive Kiran were selected to study transcriptional and physiological alterations at the same stress durations. Physiological parameters (photosynthesis rate, stomatal conductance, transpiration), and non-enzymatic antioxidants (carotenoids, anthocyanins, total phenol content) production indicated better performance of Heena than Kiran. Comparatively higher accumulation of carotenoid and anthocyanin content and the increased photosynthetic rate was also observed in Heena. Root morphology (length, numbers of root hairs, seminal roots and adventitious roots) and anatomical data (lignin deposition, xylem area) enable tolerant variety Heena to better maintain membrane integrity and relative water content, which also contribute to comparatively higher biomass accumulation in Heena under drought. In transcriptome profiling, significant drought stress-associated differentially expressed genes (DEGs) were identified in both the varieties. A total of 1033 and 936 uniquely upregulated DEGs were found in Heena and Kiran respectively. The significant modulation of DEGs that were mainly associated with phytohormone signaling, stress-responsive genes (LEA, DREB), transcription factors (TFs) (AP2/ERF, MYB, WRKY, bHLH), and genes involved in photosynthesis and antioxidative mechanisms indicate better adaptive nature of Heena in stress tolerance. Additionally, the QTL-mapping analysis showed a very high number of DEGs associated with drought stress at AQHP069 QTL in Heena in comparison to Kiran which further distinguishes the drought-responsive traits at the chromosomal level in both the contrasting varieties. Overall, results support the higher capability of Heena over Kiran variety to induce numerous genes along with the development of better root architecture to endure drought stress.

Auxin-salicylic acid cross-talk ameliorates OsMYB-R1 mediated defense towards heavy metal, drought and fungal stress.

The MYB TF family is an immensely large and functionally diverse class of proteins involved in the regulation of cell cycle, cell morphogenesis to stress signaling mechanism. The present study deciphered the hormonal cross-talk of wound inducible and stress-responsive OsMYB-R1 transcription factor in combating abiotic [Cr(VI) and drought/PEG] as well as biotic (Rhizoctonia solani) stress. OsMYB-R1 over-expressing rice transgenics exhibit a significant increase in lateral roots, which may be associated with increased tolerance under Cr(VI) and drought exposure. In contrast, its loss-of-function reduces stress tolerance. Higher auxin accumulation in the OsMYB-R1 over-expressed lines further strengthens the protective role of lateral roots under stress conditions. RNA-seq. data reveals over-representation of salicylic acid signaling molecule calcium-dependent protein kinases, which probably activate the stress-responsive downstream genes (Peroxidases, Glutathione S-transferases, Osmotins, Heat Shock Proteins, Pathogenesis Related-Proteins). Enzymatic studies further confirm OsMYB-R1 mediated robust antioxidant system as catalase, guaiacol peroxidase and superoxide dismutase activities were found to be increased in the over-expressed lines. Our results suggest that OsMYB-R1 is part of a complex network of transcription factors controlling the cross-talk of auxin and salicylic acid signaling and other genes in response to multiple stresses by modifying molecular signaling, internal cellular homeostasis and root morphology.

Self-cleansing properties of Ganga during mass ritualistic bathing on Maha-Kumbh.

The deterioration of water quality of river Ganga is a huge concern for Govt. of India. Apart from various pollution sources, the religious and ritualistic activities also have a good share in deteriorating Ganga water quality. Thus, the aim of the present study was to evaluate the changes in physico-chemical properties, microbial diversity and role of bacteriophages in controlling bacterial population of Ganga water during mass ritualistic bathing on the occasion of Maha-Kumbh in 2013. The BOD, COD, hardness, TDS and level of various ions significantly increased, while DO decreased in Ganga water during Maha-Kumbh. Ganga water was more affluent in trace elements than Yamuna and their levels further increased during Maha-Kumbh, which was correlated with decreased level of trace elements in the sediment. The bacterial diversity and evenness were increased and correlated with the number of devotees taking a dip at various events. Despite enormous increase in bacterial diversity during mass ritualistic bathing, the core bacterial species found in pre-Kumbh Ganga water were present in all the samples taken during Kumbh and post-Kumbh. In addition, the alteration in bacterial population during mass bathing was well under 2 log units which can be considered negligible. The study of bacteriophages at different bathing events revealed that Ganga was richer with the presence of bacteriophages in comparison with Yamuna against seven common bacteria found during the Maha-Kumbh. These bacteriophages have played a role in controlling bacterial growth and thus preventing putrefaction of Ganga water. Further, the abundance of trace elements in Ganga water might also be a reason for suppression of bacterial growth. Thus, the current study showed that Ganga has characteristic water quality in terms of physico-chemical property and microbial diversity that might have a role in the reported self-cleansing property of Ganga; however, the increased pollution load has surpassed its self-cleansing properties. Since water has been celebrated in all cultures, the outcome of the current study will not only be useful for the policy maker of cleaning and conservation of Ganga but also for restoration of other polluted rivers all over the world.

Structural characterization and in-silico analysis of Momordica charantia 7S globulin for stability and ACE inhibition.

Momordica charantia (Mc) seeds are widely used edible crop with high nutritional quality. The food and pharmaceutical industries use it as a natural anti-oxygenic agent. Herein, a ~52 kDa protein, which is a major part of seed proteome has been purified, biochemically characterized and structure has been determined. MALDI-ESI-MS identified peptide fragments and contig-deduced sequence suggested the protein to be homologous to 7S globulins. The crystal structure shows that protein has a bicupin fold similar to 7S globulins and the electron density for a copper and acetate ligand were observed in the C-terminal barrel domain. In silico study reveals that a tripeptide (VFK) from Mc7S possess a higher binding affinity for angiotensin converting enzyme (ACE) than already reported drug Lisinopril (LPR). The protein is a glycoprotein and highly stable under varying thermal and pH conditions due to its secondary structures. The DPPH (2,2-diphenyl-1-picryl-hydrazyl-hydrate) assay showed the protein to have an anti-oxygenic nature and can aid in scavenging free radical from sample. The protein can assist to enhance the nutritional and functional value of food by acting as a food antioxidant. Further, characterization of Mc7S required which might add in importance of Mc7S as antioxidant, anti-diabetic and anti-hypertensive.

Recent advances in arsenic metabolism in plants: current status, challenges and highlighted biotechnological intervention to reduce grain arsenic in rice.

Arsenic (As), classified as a "metalloid" element, is well known for its carcinogenicity and other toxic effects to humans. Arsenic exposure in plants results in the alteration of the physiochemical and biological properties and consequently, loss of crop yield. Being a staple food for half of the world's population, the consumption of As-contaminated rice grain by humans may pose serious health issues and risks for food security. In this study, we have described the principal understanding of the molecular basis of arsenic toxicity and accumulation in plant parts. We described the measures for decreasing As accumulation in rice and understanding the mechanism and transport of As uptake, its transport from root to shoot to rice grain, its metabolism, detoxification, as well as the mechanisms lying behind its accumulation in rice grains. There are various checkpoints, such as the tuning of AsV/Pi specific Pi transporters, arsenate reductase, transporters that are involved in the efflux of As to either the vacuole or outside the cell, xylem loading, loading and unloading to the phloem, and transporters involved in the loading of As to grain, that can be targeted to reduce As accumulation in rice grain. Genes/proteins involved in As detoxification, particularly the glutathione (GSH) biosynthesis pathway, phytochelatin (PC) synthesis, and arsenic methyltransferase, also provide a great pool of pathways that can also be castellated for the low As in rice grains. Paddy rice is also used as fodder for animals, enhancing vacuolar sequestration and using constitutive promoters, which may be of concern for animal health. Therefore, using a root-specific promoter and/or converting inorganic arsenic into volatile organic arsenic might be a better strategy for low As in grain. Furthermore, in this review, the other specific approaches, such as bio-remediation, bio-augmentation practices, and molecular breeding, which have great potential to reduce As uptake from soil to rice grains, have also been highlighted.

Unilateral renal artery stenosis presenting as acute flaccid paralysis: a rare presentation.

Renovascular hypertension is one of the common causes of secondary hypertension. Here we report a case of patient of renal artery stenosis presenting to the emergency department as a case of acute flaccid paralysis. Renal artery stenosis has been associated with hypokalaemia, but rarely reported to be symptomatic. Initial correction of hypokalaemia leads to improvement of weakness and aetiological work up for hypokalaemia with hypertension revealed hypokalaemia due to hyperaldosteronism secondary to unilateral renal artery stenosis. The patient was managed medically with aldosterone antagonist in the anti hypertensive therapy and weakness did not recur despite withdrawal of potassium supplements. On follow-up, the patient was ambulatory with no signs of weakness, controlled blood pressure and normal potassium level.

Nitric oxide mediated transcriptional modulation enhances plant adaptive responses to arsenic stress.

Arsenic (As) contamination in rice leads to yield decline and causes carcinogenic risk to human health. Although the role of nitric oxide (NO) in reducing As toxicity is known, NO-mediated genetic modulation in the plant during arsenic toxicity has not yet been established. We analyzed the key components of NO metabolism and the correlations between NO interaction and arsenic stress using rice as a relevant model plant. Illumina sequencing was used to investigate the NO-mediated genome-wide temporal transcriptomic modulation in rice root upon AsIII exposure during 12 days (d) of the growth period. Sodium nitroprusside (SNP) was used as NO donor. SNP supplementation resulted in marked decrease in ROS, cell death and As accumulation during AsIII stress. NO was found to modulate metal transporters particularly NIP, NRAMP, ABC and iron transporters, stress related genes such as CytP450, GSTs, GRXs, TFs, amino acid, hormone(s), signaling and secondary metabolism genes involved in As detoxification. We detected NO-mediated change in jasmonic acid (JA) content during AsIII stress. The study infers that NO reduces AsIII toxicity through modulating regulatory networks involved in As detoxification and JA biosynthesis.

A protective role for nitric oxide and salicylic acid for arsenite phytotoxicity in rice (Oryza sativa L.).

Nitric oxide (NO) and salicylic acid (SA) are important signaling molecules in plant system. In the present study both NO and SA showed a protective role against arsenite (AsIII) stress in rice plants when supplied exogenously. The application of NO and SA alleviated the negative impact of AsIII on plant growth. Nitric oxide supplementation to AsIII treated plants greatly decreased arsenic (As) accumulation in the roots as well as shoots/roots translocation factor. Arsenite exposure in plants decreased the endogenous levels of NO and SA. Exogenous supplementation of SA not only enhanced endogenous level of SA but also the level of NO through enhanced nitrate reductase (NR) activity, whether AsIII was present or not. Exogenously supplied NO decreased the NR activity and level of endogenous NO. Arsenic accumulation was positively correlated with the expression level of OsLsi1, a transporter responsible for AsIII uptake. The endogenous level of NO and SA were positively correlated to each other either when AsIII was present or not. This close relationship indicates that NO and SA work in harmony to modulate the signaling response in AsIII stressed plants.

Sulfur mediated reduction of arsenic toxicity involves efficient thiol metabolism and the antioxidant defense system in rice.

Arsenic (As) contamination is a global issue, with South Asia and South East Asia being worst affected. Rice is major crop in these regions and can potentially pose serious health risks due to its known As accumulation potential. Sulfur (S) is an essential macronutrient and a vital element to combat As toxicity. The aim of this study was to investigate the role of S with regards to As toxicity in rice under different S regimes. To achieve this aim, plants were stressed with AsIII and AsV under three different S conditions (low sulfur (0.5mM), normal sulfur (3.5mM) and high sulfur (5.0mM)). High S treatment resulted in increased root As accumulation, likely due to As complexation through enhanced synthesis of thiolic ligands, such as non-protein thiols and phytochelatins, which restricted As translocation to the shoots. Enzymes of S assimilatory pathways and downstream thiolic metabolites were up-regulated with increased S supplementation; however, to maintain optimum concentrations of S, transcript levels of sulfate transporters were down-regulated at high S concentration. Oxidative stress generated due to As was counterbalanced in the high S treatment by reducing hydrogen peroxide concentration and enhancing antioxidant enzyme activities. The high S concentration resulted in reduced transcript levels of Lsi2 (a known transporter of As). This reduction in Lsi2 expression level is a probable reason for low shoot As accumulation, which has potential implications in reducing the risk of As in the food chain.

Selenium ameliorates arsenic induced oxidative stress through modulation of antioxidant enzymes and thiols in rice (Oryza sativa L.).

Arsenic (As) contamination of rice is a major problem for South-East Asia. In the present study, the effect of selenium (Se) on rice (Oryza sativa L.) plants exposed to As was studied in hydroponic culture. Arsenic accumulation, plant growth, thiolic ligands and antioxidative enzyme activities were assayed after single (As and Se) and simultaneous supplementations (As + Se). The results indicated that the presence of Se (25 µM) decreased As accumulation by threefold in roots and twofold in shoots as compared to single As (25 µM) exposed plants. Arsenic induced oxidative stress in roots and shoots was significantly ameliorated by Se supplementation. The observed positive response was found associated with the increased activities of ascorbate peroxidase (APX; EC 1.11.1.11), catalase (CAT; EC 1.11.1.6) and glutathione peroxidase (GPx; EC 1.11.1.9) and induced levels of non-protein thiols (NPTs), glutathione (GSH) and phytochelatins (PCs) in As + Se exposed plants as compared to single As treatment. Selenium supplementation modulated the thiol metabolism enzymes viz., γ-glutamylcysteine synthetase (γ-ECS; EC 6.3.2.2), glutathione-S-transferase (GST; EC 2.5.1.18) and phytochelatin synthase (PCS; EC 2.3.2.15). Gene expression analysis of several metalloid responsive genes (LOX, SOD and MATE) showed upregulation during As stress, however, significant downregulation during As + Se exposure as compared to single As treatment. Gene expressions of enzymes of antioxidant and GSH and PC biosynthetic systems, such as APX, CAT, GPx, γ-ECS and PCS were found to be significantly positively correlated with their enzyme activities. The findings suggested that Se supplementation could be an effective strategy to reduce As accumulation and toxicity in rice plants.

Arsenite tolerance is related to proportional thiolic metabolite synthesis in rice (Oryza sativa L.).

Thiol metabolism is the primary detoxification strategy by which rice plants tolerate arsenic (As) stress. In light of this, it is important to understand the importance of harmonised thiol metabolism with As accumulation and tolerance in rice plant. For this aim, tolerant (T) and sensitive (S) genotypes were screened from 303 rice (Oryza sativa) genotypes on exposure to 10 and 25 µM arsenite (As(III)) in hydroponic culture. On further As accumulation estimation, contrasting (13-fold difference) T (IC-340072) and S (IC-115730) genotypes were selected. This difference was further evaluated using biochemical and molecular approaches to understand involvement of thiolic metabolism vis-a-vis As accumulation in these two genotypes. Various phytochelatin (PC) species (PC(2), PC(3) and PC(4)) were detected in both the genotypes with a dominance of PC(3). However, PC concentrations were greater in the S genotype, and it was noticed that the total PC (PC(2) + PC(3 )+ PC(4))-to-As(III) molar ratio (PC-SH:As(III)) was greater in T (2.35 and 1.36 in shoots and roots, respectively) than in the S genotype (0.90 and 0.15 in shoots and roots, respectively). Expression analysis of several metal(loid) stress-related genes showed significant upregulation of glutaredoxin, sulphate transporter, and ascorbate peroxidase in the S genotype. Furthermore, enzyme activity of phytochelatin synthase and cysteine synthase was greater on As accumulation in the S compared with the T genotype. It was concluded that the T genotype synthesizes adequate thiols to detoxify metalloid load, whereas the S genotype synthesizes greater but inadequate levels of thiols to tolerate an exceedingly greater load of metalloids, as evidenced by thiol-to-metalloid molar ratios, and therefore shows a phytotoxicity response.