Meetu Gupta

Differential response of arsenic stress in two varieties of Brassica juncea L.

Present study showed the toxicity caused by Arsenite (As(III)) and its detoxification responses in two varieties (Varuna and Pusa Bold) of Brassica juncea. Comparisons were made in leaves and roots of both the varieties, which showed that the accumulation pattern in both the varieties were dose and duration dependent, being more in roots for two days and in leaves for four days. Increase/decrease of antioxidant enzymes activities (SOD, CAT, GPX) showed not much changes at the given concentrations except that the enzyme activities showed significant increase at the lower concentrations. Semi quantitative RT-PCR analysis of PCS showed more expression of its transcript in P. Bold as compared to Varuna variety due to As(III) stress. The analysis of isoenzyme pattern in leaves of P. Bold showed five and two major bands of SOD and GPX, respectively. As(III) treatment leads to the activation of MAPK activity indicating role of this important cascade in transducing As(III) mediated signals. The data presented indicates the differential responses in both the varieties and also that the increased tolerance in P. Bold may be due to the defensive role of antioxidant enzymes, induction of MAPK and up regulation of PCS transcript which is responsible for the production of metal binding peptides.

Arsenic stress activates MAP kinase in rice roots and leaves

The toxic metalloid arsenite has become a potential threat to rice growing regions leading to serious contamination in food chain. In the present study effect of different physiological concentration of arsenite that is toxic and triggers the molecular events were evaluated in rice seedlings. Along with severe effect on the growth of rice seedling, production of reactive oxygen species (ROS) and nitric oxide (NO) in arsenite treated rice roots was also observed. Activation of a 42kDa mitogen activated protein kinase (MAPK/MPK) by arsenite was observed in rice leaves and 42 and 44kDa in roots in dose dependent manner. The activated MAPK could be immunoprecipitated with anti-phospho-tyrosine antibody, 4G10. The kinetic of MAPK activation by arsenite was found to be dose dependent. Transcript analysis of MAPK family and immunokinase assay in arsenite treated rice seedling revealed significant level of induction in OsMPK3 transcripts in leaves and OsMPK3, OsMPK4 transcripts in roots. Among MAPK kinase (MKKs) gene family, OsMKK4 transcripts were found to be induced in arsenite treated rice leaves and roots. In-silico homology modeling and docking analysis supported OsMPK3-OsMKK4 interaction. The data indicates that arsenite stress is transduced through MAPK signaling cascade in rice.

Comparative biochemical and RAPD analysis in two varieties of rice (Oryza sativa) under arsenic stress by using various biomarkers

Multiple biomarker systems have been frequently used to measure the genotoxic effects of environmental pollutants (including heavy metals) on living organisms. In this study, we used leaves of hydroponically grown 14 days old seedlings of rice (Oryza sativa) varieties (PB1 and IR64) treated with 50, 150 and 300 μM arsenite (As(III)) for 24 and 96 h duration. Reduction in seed germination, root-shoot length, chlorophyll and protein were observed with increasing As(III) concentration and duration in both varieties, being more in IR64. Increase/decrease of antioxidant enzymes and stress related parameters showed much changes at higher concentration for 24 and 96 h duration in both varieties. Eleven primers were found in RAPD analysis to produce polymorphic band pattern and produced a total of 51 (control), 79 (treated) and 42 (control) and 29 (treated) bands in PB1 and IR64 varieties, respectively. These results indicated that genomic template stability (GTS, changes in RAPD profile) was significantly affected at all tested As(III) concentration, when compared with other parameters. Differential response was observed in both varieties with PB1 being more tolerant. We concluded that DNA polymorphism detected by RAPD analysis in conjunction with other biochemical parameters could be a powerful eco-toxicological tool in bio-monitoring arsenic pollution.

An Overview of Selenium Uptake, Metabolism, and Toxicity in Plants.

Selenium (Se) is an essential micronutrient for humans and animals, but lead to toxicity when taken in excessive amounts. Plants are the main source of dietary Se, but essentiality of Se for plants is still controversial. However, Se at low doses protects the plants from variety of abiotic stresses such as cold, drought, desiccation, and metal stress. In animals, Se acts as an antioxidant and helps in reproduction, immune responses, thyroid hormone metabolism. Selenium is chemically similar to sulfur, hence taken up inside the plants via sulfur transporters present inside root plasma membrane, metabolized via sulfur assimilatory pathway, and volatilized into atmosphere. Selenium induced oxidative stress, distorted protein structure and function, are the main causes of Se toxicity in plants at high doses. Plants can play vital role in overcoming Se deficiency and Se toxicity in different regions of the world, hence, detailed mechanism of Se metabolism inside the plants is necessary for designing effective Se phytoremediation and biofortification strategies.

Selenium and auxin mitigates arsenic stress in rice (Oryza sativa L.) by combining the role of stress indicators, modulators and genotoxicity assay.

Arsenic (As) is known to disrupt the biological function in plants by inhibiting their growth and developmental process, while selenium (Se) is an essential micronutrient within the appropriate amount. Phytohormone auxin on the other hand is an established growth regulator and plays a significant role in stress management. Present study is designed to see the effect of Se and auxin on morphological and biochemical characteristics and, on the genotoxicity in rice plants under As stress. The observations indicated that seedlings supplemented only with As showed inhibition in the growth parameters, however, co-application of Se and auxin improved growth of rice seedlings, level of stress indicators, (chlorophyll, protein, MDA content) and modulators (cysteine, proline) as compared the individual treatment of As. Genomic template stability calculated through changes in RAPD profile showed consistent results when compared with the indicator and modulator parameters. Altered DNA profile showed varying degrees of polymorphism, highest in roots of As treated seedlings and lowest in roots of Se+auxin and As+Se treated seedlings. Altogether, this study conclude that application of Se and auxin alone or in combination were more effective in lowering the As induced stress in rice.

Heavy metal induced DNA changes in aquatic macrophytes: Random amplified polymorphic DNA analysis and identification of sequence characterized amplified region marker

Plants have been used as good bio-indicators and genetic toxicity of environmental pollution in recent years. In this study, aquatic plants Hydrilla verticillata and Ceratophyllum demersum treated with 10 micromol/L Cd, 5 micromol/L Hg, and 20 micromol/L Cu for 96 h, showed changes in chlorophyll, protein content, and in DNA profiles. The changes in DNA profiles included variation in band intensity, presence or absence of certain bands and even appearance of new bands. Genomic template stability test performed for the qualitative measurement of changes in randomly amplified polymorphic DNA (RAPD) profiles, showed significant effect at the given concentration of metals. Cloning and sequencing of bands suggested that these markers although may not be homologous to any known gene but its conversion as a sequence characterized amplified region (SCAR) marke is useful in detecting the effects of genotoxin agents.

Exposure of Brassica juncea (L) to arsenic species in hydroponic medium: comparative analysis in accumulation and biochemical and transcriptional alterations

Arsenic (As) contamination in the environment has attracted considerable attention worldwide. The objective of the present study was to see the comparative effect of As species As(III) and As(V) on accumulation, biochemical responses, and gene expression analysis in Brassica juncea var. Pusa Jaganath (PJn). Hydroponically grown 14-day-old seedlings of B. juncea were treated with different concentrations of As(III) and As(V). Accumulation of total As increased with increasing concentration of both As species and exposure time, mainly in roots. Reduction in seed germination, root–shoot length, chlorophyll, and protein content were observed with increasing concentration and exposure time of both As species, being more in As(III)-treated leaves. PJn variety showed that antioxidant enzymes (superoxide dismutase (SOD), catalase (CAT), and ascorbate peroxidase (APX)) and stress-related parameters (cysteine, proline, and malondialdehyde (MDA)) were stimulated and allows plant to tolerate both As species. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) analysis in leaves showed significant changes in protein profile with more stringent effect with As(III) stress. Semiquantitative RT-PCR analysis showed regulation in expression of phytochelatin synthase (PCS), metallothionine-2 (MT-2), glutathione reductase (GR), and glutathione synthetase (GS) genes under both As(III) and As(V) stresses. Results suggested that accumulation and inhibition on physiological parameters differ according to the As species, while molecular and biochemical parameters showed a combinatorial type of tolerance mechanism against As(III) and As(V) stresses.

An update on nitric oxide and its benign role in plant responses under metal stress

Pollution due to heavy metal(loid)s has become common menace across the globe. This is due to unprecedented frequent geological changes coupled with increasing anthropogenic activities, and population growth rate. Heavy metals (HMs) presence in the soil causes toxicity, and hampers plant growth and development. Plants being sessile are exposed to a variety of stress and/or a network of different kinds of stresses throughout their life cycle. To sense and transduce these stress signal, the signal reactive nitrogen species (RNS) particularly nitric oxide (NO) is an important secondary messenger next to only reactive oxygen species (ROS). Nitric oxide, a redox active molecule, colourless simple gas, and being a free radical (NO) has the potential in regulating multiple biological signaling responses in a variety of plants. Nitric oxide can counteract HMs-induced ROS, either by direct scavenging or by stimulating antioxidants defense team; therefore, it is also known as secondary antioxidant. The imbalance or cross talk of/between NO and ROS concentration along with antioxidant system leads to nitrosative and oxidative stress, or combination of both i.e., nitro-oxidative stress. Endogenous synthesis of NO also takes place in plants in the presence of heavy metals. During HM stress the different organelles of plant cells can biosynthesize NO in parallel to the ROS, such as in mitochondria, chloroplasts, peroxisomes, cytoplasm, endoplasmic reticulum and apoplasts. In view of the above, an effort has been made in the present review article to trace current knowledge and latest advances in chemical properties, biological roles, mechanism of NO action along with the physiological, biochemical, and molecular changes that occur in plants under different metal stress. A brief focus is also carried on ROS properties, roles, and their production.

Arsenate induced differential response in rice genotypes.

To study the differential response in two rice genotypes (PB1 and IR-64), hydroponically grown 14 days old plants were exposed to 50, 150 and 300µM As(V) for 24 and 96h. Accumulation of As was not significantly higher in PB1 variety except at higher concentration (300μM) and duration (96h), but up regulation of gene transcripts were higher as compared to IR-64. Inhibition in seed germination, root-shoot length, chlorophyll and protein content was observed in both varieties with increasing concentration and exposure time. PB1 variety was found more capable to detoxify As(V) through induction of antioxidant defense system and other stress related parameters (cysteine, proline content). SDS-PAGE and semi quantitative RT-PCR analysis showed significant changes in protein profile and gene expression analysis. The results suggests that various studied parameters and transcripts accumulation showed a combinatorial type of tolerance mechanism in PB1 variety to provide better protection against As(V) stress.

Mechanism of Arsenic Toxicity and Tolerance in Plants: Role of Silicon and Signalling Molecules

Arsenic (As) contamination is a major environmental problem. It has become a major limiting factor in the growth and yield of crop plants, affecting the sustainability of agriculture production. Arsenic taken up by the plant tissue causes severe damage to important cellular components, such as lipids, protein, DNA and RNA. Mostly, inorganic forms of As, arsenate and arsenite, are found to be more toxic. To mitigate and reduce the negative effects of As, various prospects have been evaluated. Silicon (Si) has been found to serve as a beneficial element for plant growth and development, and its accumulation is helpful in maintaining sustainable production. Studies have revealed the ability of Si to mitigate various biotic and abiotic stresses in crop plants. It is also known that phosphate transporter recognizes arsenate while arsenite is taken up as a Si transporter. There is a lack of information available on the interactive effects of As and Si, especially in terrestrial or crop plants. On the other hand, signalling molecules are also known to regulate plant metabolism, growth and development under various stresses. The signal pathways either operate independently or may positively or negatively modulate other pathways. This chapter examines the participation and interaction of As and Si in plants. Furthermore, role of signalling molecules is also discussed to mitigate As-induced damages.