The role of LTR retrotransposons in plant genetic engineering: how to control their transposition in the genome.

KEY MESSAGE: We briefly discuss that the similarity of LTR retrotransposons to retroviruses is a great opportunity for the development of a genetic engineering tool that exploits intragenic elements in the plant genome for plant genetic improvement. Long terminal repeat (LTR) retrotransposons are very similar to retroviruses but do not have the property of being infectious. While spreading between its host cells, a retrovirus inserts a DNA copy of its genome into the cells. The ability of retroviruses to cause infection with genome integration allows genes to be delivered to cells and tissues. Retrovirus vectors are, however, only specific to animals and insects, and, thus, are not relevant to plant genetic engineering. However, the similarity of LTR retrotransposons to retroviruses is an opportunity to explore the former as a tool for genetic engineering. Although recent long-read sequencing technologies have advanced the knowledge about transposable elements (TEs), the integration of TEs is still unable either to control them or to direct them to specific genomic locations. The use of existing intragenic elements to achieve the desired genome composition is better than using artificial constructs like vectors, but it is not yet clear how to control the process. Moreover, most LTR retrotransposons are inactive and unable to produce complete proteins. They are also highly mutable. In addition, it is impossible to find a full active copy of a LTR retrotransposon out of thousands of its own copies. Theoretically, if these elements were directly controlled and turned on or off using certain epigenetic mechanisms (inducing by stress or infection), LTR retrotransposons could be a great opportunity to develop a genetic engineering tool using intragenic elements in the plant genome. In this review, the recent developments in uncovering the nature of LTR retrotransposons and the possibility of using these intragenic elements as a tool for plant genetic engineering are briefly discussed.

Epigenetic modifications and miRNAs determine the transition of somatic cells into somatic embryos.

KEY MESSAGE: This review discusses the epigenetic changes during somatic embryo (SE) development, highlights the genes and miRNAs involved in the transition of somatic cells into SEs as a result of epigenetic changes, and draws insights on biotechnological opportunities to study SE development. Somatic embryogenesis from somatic cells occurs in a series of steps. The transition of somatic cells into somatic embryos (SEs) is the most critical step under genetic and epigenetic regulations. Major regulatory genes such as SERK, WUS, BBM, FUS3/FUSA3, AGL15, and PKL, control SE steps and development by turning on and off other regulatory genes. Gene transcription profiles of somatic cells during SE development is the result of epigenetic changes, such as DNA and histone protein modifications, that control and decide the fate of SE formation. Depending on the type of somatic cells and the treatment with plant growth regulators, epigenetic changes take place dynamically. Either hypermethylation or hypomethylation of SE-related genes promotes the transition of somatic cells. For example, the reduced levels of DNA methylation of SERK and WUS promotes SE initiation. Histone modifications also promote SE induction by regulating SE-related genes in somatic cells. In addition, miRNAs contribute to the various stages of SE by regulating the expression of auxin signaling pathway genes (TIR1, AFB2, ARF6, and ARF8), transcription factors (CUC1 and CUC2), and growth-regulating factors (GRFs) involved in SE formation. These epigenetic and miRNA functions are unique and have the potential to regenerate bipolar structures from somatic cells when a pluripotent state is induced. However, an integrated overview of the key regulators involved in SE development and downstream processes is lacking. Therefore, this review discusses epigenetic modifications involved in SE development, SE-related genes and miRNAs associated with epigenetics, and common cis-regulatory elements in the promoters of SE-related genes. Finally, we highlight future biotechnological opportunities to alter epigenetic pathways using the genome editing tool and to study the transition mechanism of somatic cells.

The plant epitranscriptome: revisiting pseudouridine and 2'-O-methyl RNA modifications.

There is growing evidence that post-transcriptional RNA modifications are highly dynamic and can be used to improve crop production. Although more than 172 unique types of RNA modifications have been identified throughout the kingdom of life, we are yet to leverage upon the understanding to optimize RNA modifications in crops to improve productivity. The contributions of internal mRNA modifications such as N6-methyladenosine (m6 A) and 5-methylcytosine (m5 C) methylations to embryonic development, root development, leaf morphogenesis, flowering, fruit ripening and stress response are sufficiently known, but the roles of the two most abundant RNA modifications, pseudouridine (Ψ) and 2'-O-methylation (Nm), in the cell remain unclear due to insufficient advances in high-throughput technologies in plant development. Therefore, in this review, we discuss the latest methods and insights gained in mapping internal Ψ and Nm and their unique properties in plants and other organisms. In addition, we discuss the limitations that remain in high-throughput technologies for qualitative and quantitative mapping of these RNA modifications and highlight future challenges in regulating the plant epitranscriptome.

Heavy metal induced regulation of plant biology: Recent insights.

The presence of different forms of heavy metals in the earth crust is very primitive and probably associated with the origin of plant life. However, since the beginning of human civilisation, heavy metal use and its contamination to all living systems on earth have significantly increased due to human anthropogenic activities. Heavy metals are nonbiodegradable, which directly or indirectly impact photosynthesis, antioxidant system, mineral nutrition status, phytohormones and amino acid-derived molecules. Due to the toxic behaviour of some heavy metals, the endogenous status of chemical messengers like phytohormones may get significantly influenced, leading to harmful impacts on plant growth, development and overall yield of the plants. It has been noticed that exogenous application of phytohormones, that is, abscisic acid, salicylic acid, auxins, brassinosteroids, cytokinins, ethylene and gibberellins can positively regulate the heavy metal toxicity in plants through the regulation of the ascorbate-glutathione cycle, nitrogen metabolism, proline metabolisms, transpiration rate, and cell division. Furthermore, it may also restrict the entry of heavy metals into the plant cells, which aids in the recovery of plant growth and productivity. Besides these, some defence molecules also assist the plant in dealing with heavy metal toxicity. Therefore, the present review aims to bridge the knowledge gap in this context and present outstanding discoveries related to plant life supportive processes during stressful conditions including phytohormones and heavy metal crosstalk along with suggestions for future research in this field.

Correction: Ramakrishnan et al. The Dynamism of Transposon Methylation for Plant Development and Stress Adaptation. Int. J. Mol. Sci. 2021, 22, 11387.

The authors would like to make the following corrections to the original publication [...].

Physiological and biochemical responses of soybean plants inoculated with Arbuscular mycorrhizal fungi and Bradyrhizobium under drought stress.

BACKGROUND: The present study aims to study the effects of biofertilizers potential of Arbuscular Mycorrhizal Fungi (AMF) and Bradyrhizobium japonicum (B. japonicum) strains on yield and growth of drought stressed soybean (Giza 111) plants at early pod stage (50 days from sowing, R3) and seed development stage (90 days from sowing, R5). RESULTS: Highest plant biomass, leaf chlorophyll content, nodulation, and grain yield were observed in the unstressed plants as compared with water stressed-plants at R3 and R5 stages. At soil rhizosphere level, AMF and B. japonicum treatments improved bacterial counts and the activities of the enzymes (dehydrogenase and phosphatase) under well-watered and drought stress conditions. Irrespective of the drought effects, AMF and B. japonicum treatments improved the growth and yield of soybean under both drought (restrained irrigation) and adequately-watered conditions as compared with untreated plants. The current study revealed that AMF and B. japonicum improved catalase (CAT) and peroxidase (POD) in the seeds, and a reverse trend was observed in case of malonaldehyde (MDA) and proline under drought stress. The relative expression of the CAT and POD genes was up-regulated by the application of biofertilizers treatments under drought stress condition. Interestingly a reverse trend was observed in the case of the relative expression of the genes involved in the proline metabolism such as P5CS, P5CR, PDH, and P5CDH under the same conditions. The present study suggests that biofertilizers diminished the inhibitory effect of drought stress on cell development and resulted in a shorter time for DNA accumulation and the cycle of cell division. There were notable changes in the activities of enzymes involved in the secondary metabolism and expression levels of GmSPS1, GmSuSy, and GmC-INV in the plants treated with biofertilizers and exposed to the drought stress at both R3 and R5 stages. These changes in the activities of secondary metabolism and their transcriptional levels caused by biofertilizers may contribute to increasing soybean tolerance to drought stress. CONCLUSIONS: The results of this study suggest that application of biofertilizers to soybean plants is a promising approach to alleviate drought stress effects on growth performance of soybean plants. The integrated application of biofertilizers may help to obtain improved resilience of the agro ecosystems to adverse impacts of climate change and help to improve soil fertility and plant growth under drought stress.

Role of jasmonic acid in plants: the molecular point of view.

KEY MESSAGE: Recent updates in JA biosynthesis, signaling pathways and the crosstalk between JA and others phytohormones in relation with plant responses to different stresses. In plants, the roles of phytohormone jasmonic acid (JA), amino acid conjugate (e.g., JA-Ile) and their derivative emerged in last decades as crucial signaling compounds implicated in stress defense and development in plants. JA has raised a great interest, and the number of researches on JA has increased rapidly highlighting the importance of this phytohormone in plant life. First, JA was considered as a stress hormone implicated in plant response to biotic stress (pathogens and herbivores) which confers resistance to biotrophic and hemibiotrophic pathogens contrarily to salicylic acid (SA) which is implicated in plant response to necrotrophic pathogens. JA is also implicated in plant responses to abiotic stress (such as soil salinity, wounding and UV). Moreover, some researchers have recently revealed that JA controls several physiological processes like root growth, growth of reproductive organs and, finally, plant senescence. JA is also involved in the biosynthesis of various metabolites (e.g., phytoalexins and terpenoids). In plants, JA signaling pathways are well studied in few plants essentially Arabidopsis thaliana, Nicotiana benthamiana, and Oryza sativa L. confirming the crucial role of this hormone in plants. In this review, we highlight the last foundlings about JA biosynthesis, JA signaling pathways and its implication in plant maturation and response to environmental constraints.

The Dynamism of Transposon Methylation for Plant Development and Stress Adaptation.

Plant development processes are regulated by epigenetic alterations that shape nuclear structure, gene expression, and phenotypic plasticity; these alterations can provide the plant with protection from environmental stresses. During plant growth and development, these processes play a significant role in regulating gene expression to remodel chromatin structure. These epigenetic alterations are mainly regulated by transposable elements (TEs) whose abundance in plant genomes results in their interaction with genomes. Thus, TEs are the main source of epigenetic changes and form a substantial part of the plant genome. Furthermore, TEs can be activated under stress conditions, and activated elements cause mutagenic effects and substantial genetic variability. This introduces novel gene functions and structural variation in the insertion sites and primarily contributes to epigenetic modifications. Altogether, these modifications indirectly or directly provide the ability to withstand environmental stresses. In recent years, many studies have shown that TE methylation plays a major role in the evolution of the plant genome through epigenetic process that regulate gene imprinting, thereby upholding genome stability. The induced genetic rearrangements and insertions of mobile genetic elements in regions of active euchromatin contribute to genome alteration, leading to genomic stress. These TE-mediated epigenetic modifications lead to phenotypic diversity, genetic variation, and environmental stress tolerance. Thus, TE methylation is essential for plant evolution and stress adaptation, and TEs hold a relevant military position in the plant genome. High-throughput techniques have greatly advanced the understanding of TE-mediated gene expression and its associations with genome methylation and suggest that controlled mobilization of TEs could be used for crop breeding. However, development application in this area has been limited, and an integrated view of TE function and subsequent processes is lacking. In this review, we explore the enormous diversity and likely functions of the TE repertoire in adaptive evolution and discuss some recent examples of how TEs impact gene expression in plant development and stress adaptation.

Seed priming and foliar application with jasmonic acid enhance salinity stress tolerance of soybean (Glycine max L.) seedlings.

BACKGROUND: Jasmonic acid (JA) is an important molecule that has a regulatory effect on many physiological processes in plant growth and development under abiotic stress. This study investigated the effect of 60 µmol L-1 of JA in seed priming (P) at 15 °C in darkness for 24 h, foliar application (F), and/or their combination effect (P + F) on two soybean cultivars - 'Nannong 99-6' (salt tolerant) and 'Lee 68' (salt sensitive) - under salinity stress (100 mmol L-1 sodium chloride (NaCl)). RESULTS: Salinity stress reduced seedling growth and biomass compared with that in the control condition. Priming and foliar application with JA and/or their combination significantly improved water potential, osmotic potential, water use efficiency, and relative water content of both cultivars under salinity stress. Similarly, seed priming with JA, foliar application of JA, and/or their combination significantly improved the following properties under salinity stress compared with the untreated seedlings: net photosynthetic rate by 68.03%, 59.85%, and 76.67% respectively; transpiration rate by 74.85%, 55.10%, and 80.26% respectively; stomatal conductance by 69.88%, 78.25%, and 26.24% respectively; intercellular carbon dioxide concentration by 61.64%, 40.06%, and 65.79% respectively; and total chlorophyll content by 47.41%, 41.02%, and 55.73% respectively. Soybean plants primed, sprayed with JA, or treated with their combination enhanced the chlorophyll fluorescence, which was damaged by salinity stress. JA treatments improved abscisic acid, gibberellic acid, and JA levels by 60.57%, 62.50% and 52.25% respectively under salt stress compared with those in the control condition. The transcriptional levels of the FeSOD, POD, CAT, and APX genes increased significantly in the NaCl-stressed seedlings irrespective of JA treatments. Moreover, JA treatment resulted in a reduction of sodium ion concentration and an increase of potassium ion concentrations in the leaf and root of both cultivars regardless of salinity stress. Monodehydroascorbate reductase, dehydroascorbate reductase, and proline contents decreased in the seedlings treated with JA under salinity stress, whereas the ascorbate content increased with JA treatment combined with NaCl stress. CONCLUSION: The application of 60 µmol L-1 JA improved plant growth by regulating the interaction between plant hormones and hydrogen peroxide, which may be involved in auxin signaling and stomatal closure under salt stress. These methods could efficiently protect early seedlings and alleviate salt stress damage and provide possibilities for use in improving soybean growth and inducing tolerance against excessive soil salinity. © 2020 Society of Chemical Industry.

Nitric oxide-mediated regulation of oxidative stress in plants under metal stress: a review on molecular and biochemical aspects.

Given their sessile nature, plants continuously face unfavorable conditions throughout their life cycle, including water scarcity, extreme temperatures and soil pollution. Among all, metal(loid)s are one of the main classes of contaminants worldwide, posing a serious threat to plant growth and development. When in excess, metals which include both essential and non-essential elements, quickly become phytotoxic, inducing the occurrence of oxidative stress. In this way, in order to ensure food production and safety, attempts to enhance plant tolerance to metal(loid)s are urgently needed. Nitric oxide (NO) is recognized as a signaling molecule, highly involved in multiple physiological events, like the response of plants to abiotic stress. Thus, substantial efforts have been made to assess NO potential in alleviating metal-induced oxidative stress in plants. In this review, an updated overview of NO-mediated protection against metal toxicity is provided. After carefully reviewing NO biosynthetic pathways, focus was given to the interaction between NO and the redox homeostasis followed by photosynthetic performance of plants under metal excess.

Current Scenario of Pb Toxicity in Plants: Unraveling Plethora of Physiological Responses.

Lead (Pb) is an extremely toxic metal for all living forms including plants. It enters plants through roots from soil or soil solution. It is considered as one of the most eminent examples of anthropogenic environmental pollutant added in environment through mining and smelting of lead ores, coal burning, waste from battery industries, leaded paints, metal plating, and automobile exhaust. Uptake of Pb in plants is a nonselective process and is driven by H+/ATPases. Translocation of Pb metal ions occurs by apoplastic movement resulting in deposition of metal ions in the endodermis and is further transported by symplastic movement. Plants exposed to high concentration of Pb show toxic symptoms due to the overproduction of reactive oxygen species (ROS) through Fenton-Haber-Weiss reaction. ROS include superoxide anion, hydroxyl radical, and hydrogen peroxide, which reach to macro- and micro-cellular levels in the plant cells and cause oxidative damage. Plant growth and plethora of biochemical and physiological attributes including plant growth, water status, photosynthetic efficiency, antioxidative defense system, phenolic compounds, metal chelators, osmolytes, and redox status are adversely influenced by Pb toxicity. Plants respond to toxic levels of Pb in varied ways such as restricted uptake of metal, chelation of metal ions to the root endodermis, enhancement in activity of antioxidative defense, alteration in metal transporters expression, and involvement of plant growth regulators.

Identification and expression analysis of auxin-responsive GH3 family genes in Chinese hickory (Carya cathayensis) during grafting.

The GH3 genes play vital roles in auxin homeostasis by conjugating excess auxin to amino acids. However, how GH3 genes function during grafting in Chinese hickory (Carya cathayensis) is largely unknown. Here, based on the transcriptome database, a comprehensive identification and expression profiling analysis of 12 GH3 genes in Chinese hickory were performed. Phylogenetic analysis indicated that CcGH3-x exists in a specific subfamily. To understand the roles of CcGH3 genes, tissue-specific expression and the response to different phytohormones were determined. Expression profiles of GH3 genes of Chinese hickory during grafting were analysed. The data suggested that 10 CcGH3 genes were down-regulated at an early stage of grafting, indicating that auxin homeostasis regulated by the CcGH3 family might be inhibited at initial stages. At the completion of grafting, expression levels of members of the CcGH3 family were restored to normal levels. Endogenous auxin levels were also measured, and the data showed that free auxin decreased to the lowest level at an early stage of grafting, and then increased during grafting. Auxin amino acid conjugation increased at an early stage of grafting in rootstock, and then decreased with progression of the graft union. Our results demonstrate that the reduced expression of CcGH3 family genes during grafting might contribute to the release of free auxin, making an important contribution to the recovery of auxin levels after grafting.

The Role of Salicylic Acid in Plants Exposed to Heavy Metals.

Salicylic acid (SA) is a very simple phenolic compound (a C7H6O3 compound composed of an aromatic ring, one carboxylic and a hydroxyl group) and this simplicity contrasts with its high versatility and the involvement of SA in several plant processes either in optimal conditions or in plants facing environmental cues, including heavy metal (HM) stress. Nowadays, a huge body of evidence has unveiled that SA plays a pivotal role as plant growth regulator and influences intra- and inter-plant communication attributable to its methyl ester form, methyl salicylate, which is highly volatile. Under stress, including HM stress, SA interacts with other plant hormones (e.g., auxins, abscisic acid, gibberellin) and promotes the stimulation of antioxidant compounds and enzymes thereby alerting HM-treated plants and helping in counteracting HM stress. The present literature survey reviews recent literature concerning the roles of SA in plants suffering from HM stress with the aim of providing a comprehensive picture about SA and HM, in order to orientate the direction of future research on this topic.

Pollution assessment and spatial distribution of roadside agricultural soils: a case study from India.

The present work studied the pH, organic carbon, phosphorus (P), calcium (Ca), magnesium (Mg), and heavy metals Cu, Cr, Co and Pb in roadside agricultural soils of Jalandhar environs of Punjab, India. A total of 120 samples in triplicates were collected from different sites for assessment of heavy metal pollution. The mean values of Cu, Cr, Co and Pb were found below the permissible limits of Indian and Swedish soil limits. Principal component analysis (PCA) showed that heavy metals have different sources of origin. The results of contamination factor (CF), geoaccumulation index (Igeo), degree of contamination (Cd) and potential ecological risk index (RI) showed low contamination and ecological risks of heavy metals in roadside agricultural soils, respectively. The maps of spatial analysis indicated that northern region of the study area is more polluted.

Castasterone attenuates insecticide induced phytotoxicity in mustard.

In the current investigation, we studied role of castasterone (CS), (a bioactive brassinosteroid) in Brassica juncea grown under imidacloprid (IMI) stress. We observed that CS-seed treatment resulted in the recovery of seedling growth under IMI toxicity. Seed treatment with CS, significantly enhanced the contents of pigments like chlorophylls, carotenoids, anthocyanins and xanthophylls under stress. Oxidative stress generated by the production of reactive oxygen species (ROS) like hydrogen peroxide and superoxide anion, was reduced after CS treatment under IMI toxicity. Antioxidative defense system got activated after CS-seed treatment, resulting in the increased activities of enzymes. Moreover, CS-seed treatment under IMI stress also stimulated the biosynthesis of organic acids of Krebs cycle (citrate, succinate, fumarate and malate) and phenolics. We also noticed that CS is also involved in the regulation of the gene expression of some key enzymes involved in pigment metabolism (CHLASE, PSY, CHS), carbon fixation (RUBISCO), Krebs cycle (CS, SUCLG1, SDH, FH), ROS generation (RBO), antioxidative enzymes (SOD, CAT, POD, DHAR, GR, GST), phenolic biosynthesis (PAL) and pesticide detoxification system (CXE, P450, NADH). This modulated gene expression after CS-treatment activated the insecticide detoxification, leading to the reduction of IMI residues. Data analysis using multivariate statistical technique i.e. multiple linear regression, also supported the fact that CS can efficiently reduce IMI induced phytotoxicity in B. juncea.

Response of Phenylpropanoid Pathway and the Role of Polyphenols in Plants under Abiotic Stress.

Phenolic compounds are an important class of plant secondary metabolites which play crucial physiological roles throughout the plant life cycle. Phenolics are produced under optimal and suboptimal conditions in plants and play key roles in developmental processes like cell division, hormonal regulation, photosynthetic activity, nutrient mineralization, and reproduction. Plants exhibit increased synthesis of polyphenols such as phenolic acids and flavonoids under abiotic stress conditions, which help the plant to cope with environmental constraints. Phenylpropanoid biosynthetic pathway is activated under abiotic stress conditions (drought, heavy metal, salinity, high/low temperature, and ultraviolet radiations) resulting in accumulation of various phenolic compounds which, among other roles, have the potential to scavenge harmful reactive oxygen species. Deepening the research focuses on the phenolic responses to abiotic stress is of great interest for the scientific community. In the present article, we discuss the biochemical and molecular mechanisms related to the activation of phenylpropanoid metabolism and we describe phenolic-mediated stress tolerance in plants. An attempt has been made to provide updated and brand-new information about the response of phenolics under a challenging environment.

Modulation of antioxidative defense expression and osmolyte content by co-application of 24-epibrassinolide and salicylic acid in Pb exposed Indian mustard plants.

The study focuses on potential of combined pre-soaking treatment of 24-Epibrassinolide (EBL) and Salicylic acid (SA) in alleviating Pb phytotoxicity in Brassica juncea L. plants. The seeds after treatment with combination of both the hormones were sown in mixture of soil, sand and manure (3:1:1) and were exposed to Pb concentrations (0.25mM, 0.50mM and 0.75mM). After 30 days of growth, the plants were harvested and processed, for quantification of various metabolites. It was found that pre-sowing of seeds in combination of EBL and SA, mitigated the adverse effects of metal stress by modulating antioxidative defense response and enhanced osmolyte contents. Dry matter content and heavy metal tolerance index were enhanced in response to co-application of EBL and SA. The levels of superoxide anions, hydrogen peroxide and malondialdehyde were lowered by the combined treatment of hormones. Enhancement in activities of guaiacol peroxidase, catalase, glutathione reductase and glutathione-s-transferase were recorded. Contents of glutathione, tocopherol and ascorbic acid were also enhanced in response to co-application of both hormones. Expression of POD, CAT, GR and GST1 genes were up-regulated whereas SOD gene was observed to be down-regulated. Contents of proline, trehalose and glycine betaine were also reported to be elevated as a result of treatment with EBL+SA. The results suggest that co-application of EBL+SA may play an imperative role in improving the antioxidative defense expression of B. juncea plants to combat the oxidative stress generated by Pb toxicity.

Role of 24-epibrassinolide (EBL) in mediating heavy metal and pesticide induced oxidative stress in plants: A review.

Industrialization and urbanization have posed serious threats to the environment. Excessive release of heavy metals from industrial effluents and overuse of pesticides in modern agriculture are limiting crop production by polluting environment and deteriorating food quality. Sustaining food quality under heavy metals and pesticide stress is crucial to meet the increasing demands for food. 24-Epibrassinolide (EBL), a ubiquitously occurring plant growth hormone shows great potential to alleviate heavy metals and pesticide stress in plants. This review sums up the potential role of EBL in ameliorating heavy metals and pesticide toxicity in plants extensively. EBL application increases plant's overall growth, biomass accumulation and photosynthetic efficiency by the modulation of numerous biochemical and physiological processes under heavy metals and pesticide stress. In addition, EBL scavenges reactive oxygen species (ROS) by triggering the production of antioxidant enzymes such as SOD, CAT, POX etc. EBL also induces the production of proline and soluble proteins that helps in maintaining osmotic potential and osmo-protection under both heavy metals and pesticide stress. At the end, future needs of research about the application of 24-epibrassinolide have also been discussed.

Castasterone confers copper stress tolerance by regulating antioxidant enzyme responses, antioxidants, and amino acid balance in B. juncea seedlings.

The aim of the present study was to explore the effect of exogenous application of castasterone (CS) on physiologic and biochemical responses in Brassica juncea seedlings under copper (Cu) stress. Seeds were pre-soaked in different concentrations of CS and grown for 7 days under various levels of Cu. The exposure of B. juncea to higher levels of Cu led to decrease of morphologic parameters, with partial recovery of length and fresh weight in the CS pre-treated seedlings. Metal content was high in both roots and shoots under Cu exposure while the CS pre-treatment reduced the metal uptake. Accumulation of hydrogen peroxide (H2O2) and superoxide anion radical (O2-) were chosen as stress biomarker and higher levels of H2O2 (88.89%) and O2- (62.11%) showed the oxidative stress in metal treated B. juncea seedlings, however, CS pre-treatment reduced ROS accumulation in Cu-exposed seedlings. The Cu exposures lead to enhance the plant's enzymatic and non-enzymatic antioxidant system. It was observed that enzymatic activities of ascorbate peroxidase (APOX), dehydroascorbate reductase (DHAR), and glutathione reductase (GR), glutathione perxoidase (GPOX) and gultrathione-s-transferase increased while activity of monodehydroascorbate reductase (MDHAR) decreased under Cu stress. The pre-treatment with CS positively affected the activities of enzymes. RT-PCR analysis showed that mRNA transcript levels were correlated with total enzymatic activity of DHAR, GR, GST and GSH. Increase in the gene expression of DHAR (1.85 folds), GR (3.24 folds), GST-1 (2.00 folds) and GSH-S (3.18 folds) was noticed with CS pre-treatment. Overall, the present study shows that Cu exposure induced severe oxidative stress in B. juncea plants and exogenous application of CS improved antioxidative defense system by modulating the ascorbate-glutathione cycle and amino acid metabolism.

24-epibrassinolide stimulates imidacloprid detoxification by modulating the gene expression of Brassica juncea L.

BACKGROUND: Pesticides cause oxidative stress to plants and their residues persist in plant parts, which are a major concern for the environment as well as human health. Brassinosteroids (BRs) are known to protect plants from abiotic stress conditions including pesticide toxicity. The present study demonstrated the effects of seed-soaking with 24-epibrassinolide (EBR) on physiological responses of 10-day old Brassica juncea seedlings grown under imidacloprid (IMI) toxicity. RESULTS: In the seedlings raised from EBR-treated seeds and grown under IMI toxicity, the contents of hydrogen peroxide (H2O2) and superoxide anion (O.2-) were decreased, accompanied by enhanced activities of superoxide dismutase (SOD), catalase (CAT), glutathione reductase (GR), glutathione-S-transferase (GST), guaiacol peroxidase (POD) and the content of glutathione (GSH). As compared to controls, the gene expressions of SOD, CAT, GR, POD, NADH (NADH-ubiquinone oxidoreductase), CXE (carboxylesterase), GSH-S (glutathione synthase), GSH-T (glutathione transporter-1), P450 (cytochrome P450 monooxygenase) and GST1-3,5-6 were enhanced in the seedlings raised from EBR-treated seeds and grown in IMI supplemented substratum. However, expression of RBO (respiratory burst oxidase, the gene responsible for H2O2 production) was decreased in seedlings raised from EBR treated seeds and grown under IMI toxicity. Further, the EBR seed treatment decreased IMI residues by more than 38% in B. juncea seedlings. CONCLUSIONS: The present study revealed that EBR seed soaking can efficiently reduce oxidative stress and IMI residues by modulating the gene expression of B. juncea under IMI stress. In conclusion, exogenous EBR application can protect plants from pesticide phytotoxicity.