Prakash M. Gopalakrishnan Nair

Study on the correlation between copper oxide nanoparticles induced growth suppression and enhanced lignification in Indian mustard (Brassica juncea L.).

In this study, the morphological, physiological and molecular level effects of copper oxide nanoparticles (CuONPs) were studied in an economically important oil seed crop Brassica juncea L. The possible involvement of lignification on shoot-root growth retardation was also studied. The seedlings were exposed to 0, 20, 50, 100, 200, 400 and 500mg/L of CuONPs in semi-solid half strength Murashige and Skoog medium under controlled growth chamber conditions for 14 days. Exposure to CuONPs resulted in suppression of shoot-growth, reduction in total chlorophyll and carotenoids contents as well modification of root system architecture such as shortening of primary and lateral roots. Significant increases in hydrogen peroxide formation, peroxidase enzyme activity and lignification of shoots and roots were observed. The lipid peroxidation levels increased significantly in shoots and roots of B. juncea seedlings. Phloroglucinol-HCl staining revealed enhanced lignification of shoot and roots. Gene expression studies revealed significant activation of CuZn superoxide dismutase (CuZnSOD) in roots at all concentrations of CuONPs exposure. In shoots significant up-regulation of CuZnSOD gene was observed upon exposure to 100, 200 and 400 mg/L of CuONPs exposure. However no change in the expression levels of MnSOD gene was observed in both stem and roots. The expression of catalase (CAT) and ascorbate peroxidase (APX) genes were also not changed in shoots. However, significant inhibition of CAT and APX genes were observed in roots of B. juncea plants under exposure to 100, 200, 400 and 500 mg/L of CuONPs exposure. The SOD enzyme activity significantly increased in roots under exposure to 50-500 mg/L of CuONPs and in shoots as a result of exposure to 100-500 mg/L of CuONPs. The APX activity significantly decreased in roots upon exposure to 50-500 mg/L of CuONPs. In shoots, the APX activity significantly decreased upon exposure to 200-500 mg/L of CuONPs.

Regulation of morphological, molecular and nutrient status in Arabidopsis thaliana seedlings in response to ZnO nanoparticles and Zn ion exposure

This study examined the mechanism of toxicity in Arabidopsis thaliana seedlings to zinc oxide nanoparticles (ZnO NPs) and zinc (Zn) ions. We subjected plants to different ZnO NPs and Zn ion concentrations (0, 20, 50, 100 and 200mg/L) and analyzed resulting morphological changes, transcriptional regulation of genes involved in Zn-homeostasis, macro- and microelement homeostasis, as well as auxin regulation. Except for 20mg/L, the fresh weight and primary root length was reduced after exposure to all other concentrations of Zn ion and ZnO NP concentrations. An increase in lateral root formation (19 and 32%) was observed after exposure to 20 and 50mg/L of Zn ions respectively; whereas 20mg/L ZnO NPs treatment triggered a 9% increase in lateral root formation. Both qualitative, using Zynpyr-1 fluorescent probe and quantitative analysis revealed Zn uptake and translocation from roots to shoots after Zn ion exposure. However, ZnO NPs-treated seedlings resulted in no root to shoot translocation and Zn accumulation was mainly located in root tips, primary-lateral root junctions and root- shoot junctions. The macronutrients viz. P (1.34mg/kg DW), K (13.29mg/kg DW), S (1.29mg/kg DW) and micronutrients Cu (0.004mg/kg DW) and Fe (0.345mg/kg DW) contents were highly decreased as a result of exposure to 200mg/L of Zn ions. Similarly, the highest reduction of P (2.30mg/kg DW), K (6.36mg/kg DW), S (2.63mg/kg DW) and Cu (0.004mg/kg DW) was observed after exposure to 200mg/L of ZnO NPs. Gene regulation studies indicated the transcriptional modulation of various genes involved in Zn, macro- and micro nutrient homeostasis as well as hormone regulation. Taken together, it was observed that the mechanism of toxicity of Zn ions and ZnO NPs were different. These findings will help to design safer strategies for the application of ZnO NPs as plant fertilizer without compromising the morphological and nutritional qualities as well as for the future phytoremediation purposes.

Alteration in the expression of antioxidant and detoxification genes in Chironomus riparius exposed to zinc oxide nanoparticles.

Zinc oxide nanoparticles (ZnONPs) are widely used in several commercial products due to their unique physicochemical properties. However, their release into the aquatic environments through various anthropogenic activities will lead to toxic effect in aquatic organisms. Although several investigations have been reported on the effect of ZnONPs in aquatic organisms using traditional end points such as survival, growth, and reproduction, the molecular level end points are faster and sensitive. In this study, the expression of different genes involved in oxidative stress response, detoxification, and cellular defense was studied in an ecotoxicologically important bio-monitoring organism Chironomus riparius in order to understand the subcellular effects of ZnONPs. The fourth instar larvae were exposed to 0, 0.2, 2, 10, and 20 mg/L of ZnONPs and Zn ions (in the form of ZnSO4.7H2O) for 24 and 48 h period. The expression of CuZn superoxide dismutase, manganese superoxide dismutase, catalase, phospholipid hydroperoxide glutathione peroxidase, thioredoxin reductase 1 and delta-3, sigma-4 and epsilon-1 classes of glutathione S-transferases, cytochrome p4509AT2, and heat shock protein 70 were studied using real-time polymerase chain reaction method. Gene expression results showed that the expression of genes related to oxidative stress response was more pronounced as a result of ZnONPs exposure as compared to Zn ions. The mRNA expression of genes involved in detoxification and cellular protection was also modulated. Significantly higher expression levels of oxidative stress-related genes shows that oxidative stress is an important mechanism of toxicity as a result of ZnONPs exposure in C. riparius.

Biochemical, anatomical and molecular level changes in cucumber (Cucumis sativus) seedlings exposed to copper oxide nanoparticles

Abstract The effect of copper oxide nanoparticles (CuONPs) at the biochemical, anatomical and molecular level was investigated in cucumber (Cucumis sativus L.) seedlings. The seedlings were grown in semi-solid half strength Murashige and Skoog medium supplemented with 0, 50, 100, 200, 400 and 500 mg/L of CuONPs for fifteen days under controlled growth chamber conditions. The results showed that exposure to all concentrations of CuONPs resulted in significant reduction in shoot and root growth and biomass. A concentration-dependant increase in reactive oxygen species (ROS) generation, malondialdehyde production, lignin content and decline in mitochondrial membrane potential were observed. Cross-sections of stem showed anatomical changes, viz. an increase in xylogenesis in CuONPs exposed plants. Significant modulation in the expression of genes related to oxidative stress and lignin biosynthesis, i.e. catalase, ascorbate peroxidase, phenylalanine ammonia lyase, cinnamate 4-hydroxylase, anionic and cationic peroxidases were observed in shoots and roots as a result of CuONPs exposure. Taken together, exposure to CuONPs has resulted in excess ROS generation, lignification and growth suppression in Cucumis sativus seedlings.

Toxicological Impact of Carbon Nanomaterials on Plants

The fast growth of nanotechnology has resulted in the production and use of engineered nanoparticles with unique physical and chemical properties in various fields. The increased utilization of engineered nanoparticles enhances the risks associated with their release into the environment. The smaller size and modified physico-chemical properties raise concerns about their entry and adverse effects in plants. For instance, studies have shown that nanomaterials can be absorbed and translocated within plants. Since plants represent a major component of the ecosystem, the accumulation of engineered nanoparticles in plants is a threat to plants and the food chain.