Metabolomic profiling reveals key factors and associated pathways regulating the differential behavior of rice (Oryza sativa L.) genotypes exposed to geogenic arsenic.

Arsenic (As) toxicity is an escalating problem; however, information about the metabolic events controlling the varied pattern of As accumulation in rice genotypes within their natural environment is still lacking. The present study is thus an advancement in unravelling the response of such rice genotypes. Soil-water-rice samples were analyzed for As accumulation using ICP-MS. Furthermore, we implemented metabolomics through LC-MS/MS and UHPLC to identify metabolic signatures regulating As content by observing the metalloid's composition in rice agrosystem. Results showed that rice genotypes differed significantly in their levels of metabolites, with Mini mansoori and Pioneer having the highest levels. Mini mansoori contained least As which might have been regulated by Ala, Ser, Glu, Phe, Asn, His, Ile, Lys, Gln, Trp, Tyr, chlorogenic, p-coumaric, trans-ferulic, rutin, morin, naringenin, kampferol, and myricetin, while Asp, Arg, Met, syringic, epigalocatechin, and apigenin contributed to the greater As acclimatization ability of Pioneer. Multivariate tools separated the rice genotypes into two major clusters: Pioneer-Mini mansoori and Damini-Sampoorna-Chintu. KEGG identified three major metabolic pathways (aminoacyl-tRNA, phenylpropanoid, and secondary metabolites biosynthesis route) linked with As tolerance and adaptation mechanisms in rice. Overall, these two genotypes symbolize their As hostile and accommodating attitudes probably due to the accumulated metabolites and the physicochemical attributes of the soil-water. Thus, thorough understanding of the metabolic reactions to As may facilitate the emergence of As tolerant/resilient genotypes. This will aid in the selection of molecular markers to cultivate healthier rice genotypes in As-contaminated areas.

Exposure of biosynthesized nanoscale ZnO to Brassica juncea crop plant: morphological, biochemical and molecular aspects.

The present work describes the in vitro synthesis and characterization of Zinc oxide nanoparticles (ZnO NPs) using an enzyme alpha amylase, the synthesized nanoparticles were used to study their beneficial effect in the growth and development of Brassica juncea. Transmission Electron Microscope (TEM) image reveals the average size of ZnO NPs was 11 nm and X-ray powder diffraction (XRD) suggests nanoparticles were crystalline in nature. In-silico study confirmed lysine, glutamine and tyrosine present in alpha amylase enzyme, plays a crucial role in the reduction of Zinc acetate dihydrate to ZnO NPs. The biochemical parameters and oxidative enzymes of Brassica juncea were compared with ZnO NPs treated plants. The effect of ZnO NPs on the cellular expression of metal tolerant protein (BjMTP) and cation efflux transporter gene (BjCET2) was also studied. The results indicate that nanoparticles can be used as a replacement for traditional harmful chemical fertilizers.

Sulphur potentiates selenium to alleviate arsenic-induced stress by modulating oxidative stress, accumulation and thiol-ascorbate metabolism in Brassica juncea L.

The present study was designed to see the influence of selenium (Se) and sulphur (S) in the alleviation of arsenic (As)-induced stress in Brassica juncea plant. Se-induced alterations in physiological and biochemical responses due to deficient S (DS), normal S (NS) and additional S (AS) conditions were evaluated in 14-day-old seedlings of B. juncea variety Varuna. During the last 7 days of the 14-day-old seedlings, supplementation with arsenite (AsIII, 300 µM) alone and its combination with selenite (SeIV, 50 µM) along with different S treatments was done which are as follows: (i) control; (ii) As; (iii) As+Se+DS; (iv) As+Se + NS; (v) As+Se + AS. Experimental results showed that the application of AS in spite of NS supplied with Se influenced plant growth, oxidative stress and thiol-ascorbate-related parameters more prominently under As stress. The plants with As+Se+AS treatment exhibited lower ROS (superoxide and hydrogen peroxide ion), malondialdehyde (MDA) accumulation and lipoxygenase activity with increased activities of superoxide dismutase, catalase and ascorbate peroxidase compared with As+Se+NS condition. These plants also exhibited an increase in cysteine, non-protein thiols and phytochelatins, along with reduced, oxidised and redox content of glutathione and ascorbate. Furthermore, the application of S along with Se increased the activities of glutathione reductase, glutathione S-transferase, glutathione peroxidase, monodehydroascorbate and dehydroascorbate to minimise As stress. However, we observed that these responses were reversed under As+Se+DS condition and induced oxidative stress, which was almost similar to As only treatment. It indicated that AS nutrition potentiated Se to alleviate As-inhibited plant growth by modulating antioxidants including thiol-ascorbate-based mechanism and reducing As accumulation in B. juncea plants.

Nitric oxide and abscisic acid protects against PEG-induced drought stress differentially in Brassica genotypes by combining the role of stress modulators, markers and antioxidants.

The present study was designed to see the effect of exogenous nitric oxide (NO) and abscisic acid (ABA) and their interaction on physiological and biochemical activities in leaves and roots of two Indian mustard (Brassica juncea) cultivars [cv. Pusa Jagannath (PJN) and Varuna (VAR)] exposed to polyethylene glycol (PEG)-induced drought stress. Seven days old hydroponically grown seedlings were treated with PEG (10%), sodium nitroprusside, a NO donor [NO (100 µM)] and abscisic acid [ABA (10 µM)], using different combinations as: Control, ABA, NO, PEG, PEG + ABA, PEG + NO, and PEG + NO + ABA. Results revealed that in response to PEG-induced drought stress leaf relative water content, chlorophyll, carotenoid and protein content decreased with increased production of O2-●, MDA, H2O2, cysteine content and non-enzymatic antioxidants (including proline, flavonoid, phenolic, anthocyanin, and ascorbic acid), whereas, the enzymatic antioxidants (including SOD, CAT, APX, GR) showed the response range from no effect to increase or decrease in certain enzymes in both Brassica cultivars. The application of NO or/and ABA in PEG-stressed cultivars showed that both enzymatic and non-enzymatic antioxidants responded differently to attenuate oxidative stress in leaves and roots of both cultivars. Overall, PJN had the antioxidant protection mainly through the accumulation of non-enzymatic antioxidants, whereas VAR showed tolerance by the enhancement of both enzymatic and non-enzymatic antioxidant activities. Altogether, the study concluded that the independent NO and its interaction with ABA (PEG + NO and PEG + NO + ABA) were much effective than independent ABA (PEG + ABA) in lowering PEG-drought stress in Brassica cultivars.

Arsenic-silicon priming of rice (Oryza sativa L.) seeds influence mineral nutrient uptake and biochemical responses through modulation of Lsi-1, Lsi-2, Lsi-6 and nutrient transporter genes.

Silicon (Si) has attracted substantial attention because of its beneficial effect on plants during abiotic stress, including stress due to arsenic (As). We here report that priming rice seeds with As and Si together, helped the plant to sustain As stress for longer period. We examined Si induced tolerance against As in rice seedlings at short (7 d) and long (15 d) exposure periods under As(III) and Si treatments since their germinating stage. Results showed that the expression of As(III) transporter genes OsLsi1, OsLsi2 and OsLsi6 was more in As(III) + Si treatment as compared to control and Si treatment, but lower than As(III) alone treatments. The gene expression was maximum in shoot and root at 15 d over 7 d under both As(III) and As(III) + Si treatment, which ultimately leads to decreased accumulation of As in the presence of Si. Morphological characters, antioxidant capacity, oxidative stress marker (MDA), stress modulators (cysteine, proline), and enzymes related with ascorbate-glutathione cycle significantly altered during As(III) + Si treatment at both exposure periods. Further, macro and micronutrient contents also improved with Si, and differentially regulated 12 key genes (NR, NiR, AMT, NR, GS, GOGAT, PT, PHT1, PHT2, APase, KAT1 and HAK10) related with NPK transport and utilization. Results highlight that Si priming of seeds along with As(III) influences growth positively of As-stressed rice.

Impact of silicon on Indian mustard (Brassica juncea L.) root traits by regulating growth parameters, cellular antioxidants and stress modulators under arsenic stress.

Arsenic (As) is an emerging pollutant causing inhibition in growth and development of plants resulting into phytotoxicity. On the other hand, silicon (Si) has been suggested as a modulator in abiotic and biotic stresses that, enhances plant's physiological adaptations in response to several stresses including heavy metal stress. In this study, we used roots of hydroponically grown 14 day old seedlings of Brassica juncea var. Varuna treated with 150 µM As, 1.5 mM Si and both in combination for 96 h duration. Application of Si modulated the effect of As by improving morphological traits of root along with the development of both primary and lateral roots. Changes observed in root traits showed positive correlation with As induced cell death, accumulation of reactive oxygen species (ROS), nitric oxide (NO) and intracellular superoxide radicals (O2(-)). Addition of 1.5 mM Si during As stress increased accumulation of As in roots. Mineral nutrient analysis was done using energy-dispersive X-ray fluorescence (EDXRF) technique and positively correlated with increased cysteine, proline, MDA, H2O2 and activity of antioxidant enzymes (SOD, CAT and APX). The results obtained from the above biochemical approaches support the protective and active role of Si in the regulation of As stress through the changes in root developmental process.