Zinc oxide nanoparticles and Klebsiella sp. SBP-8 alleviates chromium toxicity in Brassica juncea by regulation of antioxidant capacity, osmolyte production, nutritional content and reduction in chromium adsorption.

Heavy metals are one of the most damaging environmental toxins that hamper growth of plants. These noxious chemicals include lead (Pb), arsenic (As), nickel (Ni), cadmium (Cd) and chromium (Cr). Chromium is one of the toxic metal which induces various oxidative processes in plants. The emerging role of nanoparticles as pesticides, fertilizers and growth regulators have attracted the attention of various scientists. Current study was conducted to explore the potential of zinc oxide nanoparticles (ZnONPs) alone and in combination with plant growth promoting rhizobacteria (PGPR) Klebsiella sp. SBP-8 in Cr stress alleviation in Brassica juncea (L.). Chromium stress reduced shoot fresh weight (40%), root fresh weight (28%), shoot dry weight (28%) and root dry weight (34%) in B. juncea seedlings. Chromium stressed B. juncea plants showed enhanced levels of malondialdehyde (MDA), electrolyte leakage (EL), hydrogen peroxide (H2O2) and superoxide ion (O2• -). However, co-supplementation of ZnONPs and Klebsiella sp. SBP-8 escalated the activity of antioxidant enzymes i.e., superoxide dismutase (SOD), catalase (CAT), ascorbate peroxidase (APX) in B. juncea grown in normal and Cr-toxic soil. It is further proposed that combined treatment of ZnONPs and Klebsiella sp. SBP-8 may be useful for alleviation of other abiotic stresses in plants.

The elemental defense effect of cadmium on Alternaria brassicicola in Brassica juncea.

BACKGROUND: The elemental defense hypothesis states a new defensive strategy that hyperaccumulators defense against herbivores or pathogens attacks by accumulating heavy metals. Brassica juncea has an excellent ability of cadmium (Cd) accumulation. However, the elemental defense effect and its regulation mechanism in B. juncea remain unclear. RESULTS: In this study, we profiled the elemental defense effect and the molecular regulatory mechanism in Cd-accumulated B. juncea after Alternaria brassicicola infection. B. juncea treated with 180 mg Kg- 1 DW CdCl2 2.5H2O exhibited obvious elemental defense effect after 72 h of infection with A. brassicicola. The expression of some defense-related genes including BjNPR1, BjPR12, BjPR2, and stress-related miRNAs (miR156, miR397, miR398a, miR398b/c, miR408, miR395a, miR395b, miR396a, and miR396b) were remarkably elevated during elemental defense in B. juncea. CONCLUSIONS: The results indicate that Cd-accumulated B. juncea may defend against pathogens by coordinating salicylic acid (SA) and jasmonic acid (JA) mediated systemic acquired resistance (SAR) and elemental defense in a synergistic joint effect. Furthermore, the expression of miRNAs related to heavy metal stress response and disease resistance may regulate the balance between pathogen defense and heavy metal stress-responsive in B. juncea. The findings provide experimental evidence for the elemental defense hypothesis in plants from the perspectives of phytohormones, defense-related genes, and miRNAs.

Leaf nodule endosymbiotic Burkholderia confer targeted allelopathy to their Psychotria hosts.

After a century of investigations, the function of the obligate betaproteobacterial endosymbionts accommodated in leaf nodules of tropical Rubiaceae remained enigmatic. We report that the α-D-glucose analogue (+)-streptol, systemically supplied by mature Ca. Burkholderia kirkii nodules to their Psychotria hosts, exhibits potent and selective root growth inhibiting activity. We provide compelling evidence that (+)-streptol specifically affects meristematic root cells transitioning to anisotropic elongation by disrupting cell wall organization in a mechanism of action that is distinct from canonical cellulose biosynthesis inhibitors. We observed no inhibitory or cytotoxic effects on organisms other than seed plants, further suggesting (+)-streptol as a bona fide allelochemical. We propose that the suppression of growth of plant competitors is a major driver of the formation and maintenance of the Psychotria-Burkholderia association. In addition to potential agricultural applications as a herbicidal agent, (+)-streptol might also prove useful to dissect plant cell and organ growth processes.

Effect of potassium fertilizer on the growth, physiological parameters, and water status of Brassica juncea cultivars under different irrigation regimes.

Abiotic stress, especially a lack of water, can significantly reduce crop yields. In this study, we evaluated the physiological and biochemical effects of potassium sulfate (K2SO4) fertilizer and varied irrigation regimes on the economically significant oilseed crop, Brassica juncea L, under open field conditions. Two cultivars (RH-725 and RH-749) of B. juncea were used in a randomized complete block design experiment with three replicates. Irrigation regimes consisted of a control (double irrigation: once at the 50% flowering and another at 50% fruiting stages), early irrigation (at 50% flowering only), late irrigation (at 50% fruiting only) and stress (no irrigation). The K2SO4 applications were: control (K0, no fertilization); K1, 10 kg ha-1; and K2, 20 kg ha-1. We measured growth via fresh and dry plant weight, plant height, root length, and leaf area. All the growth parameters were higher in RH-749. The physiological attributes, including the membrane stability index and relative water content, were higher at the 50% flowering stage in RH-749. The amount of antioxidant enzymes (catalase (CAT), guaiacol peroxidase (POX), ascorbate peroxidase (APX), and superoxide dismutase (SOD)) was enhanced when both plants were fertilized during water stress. All of these enzymes had higher activity in RH-749. The total chlorophyll content and photosynthesis rate were considerably higher in RH-749, which leaked fewer electrolytes and maintained a less destructive osmotic potential under limited water conditions. The results indicated that it is water-stress tolerant when given a high concentration of K2SO4, which alleviated the adverse effects of water stress on growth and physiology.

Root NRT, NiR, AMT, GS, GOGAT and GDH expression levels reveal NO and ABA mediated drought tolerance in Brassica juncea L.

Little is known about the interactive effects of exogenous nitric oxide (NO) and abscisic acid (ABA) on nitrogen (N) metabolism and related changes at molecular and biochemical levels under drought stress. The present study highlights the independent and combined effect of NO and ABA (grouped as "nitrate agonists") on expression profiles of representative key genes known to be involved in N-uptake and assimilation, together with proline metabolism, N-NO metabolism enzyme's activity and nutrient content in polyethylene glycol (PEG) treated roots of Indian mustard (B. juncea cv. Varuna). Here we report that PEG mediated drought stress negatively inhibited growth performance, as manifested by reduced biomass (fresh and dry weight) production. Total N content and other nitrogenous compounds (NO3-, NO2-) were decreased; however, NH4+, NH4+/ NO3- ratio and total free amino acids content were increased. These results were positively correlated with the PEG induced changes in expression of genes and enzymes involved in N-uptake and assimilation. Also, PEG supply lowered the content of macro- and micro-nutrients but proline level and the activity of ∆1-pyrroline-5-carboxylate synthetase increased indicating increased oxidative stress. However, all these responses were reversed upon the exogenous application of nitrate agonists (PEG + NO, PEG + NO + ABA, and PEG + ABA) where NO containing nitrate agonist treatment i.e. PEG + NO was significantly more effective than PEG + ABA in alleviating drought stress. Further, increases in activities of L-arginine dependent NOS-like enzyme and S-nitrosoglutathione reductase were observed under nitrate agonist treatments. This indicates that the balanced endogenous change in NO and ABA levels together during synthesis and degradation of NO mitigated the oxidative stress in Indian mustard seedlings. Overall, our results reveal that NO independently or together with ABA may contribute to improved crop growth and productivity under drought stress.

Nitric oxide induced Cd tolerance and phytoremediation potential of B. juncea by the modulation of antioxidant defense system and ROS detoxification.

The present study designed to illustrate correlation between cadmium induced stress and plant growth, photosynthetic pigments, morphological and physiological attributes. To study these parameters 2 weeks old seedling of B. juncea were subjected to 50 µM Cd, 100 µM Cd and 100 µM SNP separately and in combination with SNP. After 96 h, the treated plant were harvested to analyze the cellular homeostasis and metal tolerance mechanism via examining growth, stress parameters, enzymatic and non enzymatic antioxidants and expression level of NR. Higher level of Cd (100 µM) significantly increased accumulation of reactive oxygen species and malonaldehyde content in comparison to 50 µM Cd. Exogenous supplementation of SNP (100 µM) to 50 µM Cd treated plant had an additive effect on plant growth by improving the level of proline, photosynthetic pigments and activities of enzymatic antioxidants which was confirmed by histochemical staining for NADPH-d and NO fluorescence from DAF-DA staining in roots of B. juncea. Applying SNP to 50 µM Cd exposed B. juncea roots enhanced NR activity by 1.36 folds and increased NO production by 1.12 folds than individual Cd treated roots. In addition, semi quantitative RT-PCR study revealed the induction of BjNR was more pronounced in 50 µM Cd treated roots in comparison to 100 µM Cd treated roots. The present finding revealed NO confers increased B. juncea tolerance to Cd stress by stimulation of antioxidants and reestablishment of cellular redox status. Different biochemical analysis showed that plant growth, photosynthetic pigment and antioxidants were positively correlated with NO and it's negatively correlated with oxidative stress biomarkers. Therefore, NO is gaseous signalling molecule with potential role in Cd detoxification mechanism in B. juncea.

Silicon-induced postponement of leaf senescence is accompanied by modulation of antioxidative defense and ion homeostasis in mustard (Brassica juncea) seedlings exposed to salinity and drought stress.

Soil salinity and drought stress (DS) are the massive problem for worldwide agriculture. Both stresses together become more toxic to the plant growth and development. Silicon (Si) being the second most abundant element in the earth's crust, exerts beneficial effects on plants under both stress and non-stress conditions. However, limited information is available to substantiate the beneficial role of Si in delaying the premature leaf senescence and imparting tolerance of mustard (Brassica juncea L.) plants to salinity and DS. Therefore, the present study aimed to explore the role of Si (source K2SiO3) in chlorophyll (Chl) biosynthesis, nutrients uptake, relative water content (RWC), proline (Pro) metabolism, antioxidant system and delaying of premature leaf senescence in mustard plants under sodium chloride (NaCl) and DS conditions. Results of this study show that exogenous Si (1.7 mM) significantly delayed the salt plus DS-induced premature leaf senescence. This was further accompanied by the enhanced nutrients accumulation and activity of chlorophyll metabolizing enzymes [δ-aminolevulinic acid (δ-ALA) dehydratase and porphobilinogen deaminase] and levels of δ-ALA, and Chls a and b and also by decreased the Chl degradation and Chl degrading enzymes (Chlorophyllase, Chl-degrading peroxidase, pheophytinase) activity. Exogenous Si treatment induced redox homoeostasis in B. juncea L. plants, which is evident by a reduced generation of reactive oxygen species (ROS) resulting due to suppressed activity of their generating enzymes (glycolate oxidase and NADPH oxidase) and enhanced defence system. Furthermore, application of Si inhibited the activity of protease and triggered the activity of antioxidant enzymes (superoxide dismutase, catalase, ascorbate peroxidase and glutathione reductase) and plasma membrane H+-ATPase activity. In conclusion, all these results reveal that Si could help in the modulation of Chl metabolism, redox hemostasis, and the regulation of nutrients (nitrogen, phosphorus, Si and potassium) uptake in the mustard plants that lead to the postponement of premature leaf senescence under salinity plus DS.

5-aminolevulinic acid regulates Krebs cycle, antioxidative system and gene expression in Brassica juncea L. to confer tolerance against lead toxicity.

Heavy metal pollution seriously impairs crop production and poses serious concerns for human health. Exogenous application of biomolecules has been efficiently tested for enhancing plant resistance to metal toxicity. Current study evaluates the possible effect of 5-aminolevulinic acid (ALA) in Brassica juncea L. seedlings subjected to lead (Pb) stress. Our results showed that shoot length, root length and chlorophyll contents were significantly recovered in Pb stressed seedlings after ALA application, accompanied by reduction in the Pb accumulation. Significant reduction in the contents of reactive oxygen species (ROS) like superoxide anion, hydrogen peroxide and malondialdehyde were also observed in ALA treated seedlings under Pb stress. Furthermore, we also noticed enhancement in the activities of antioxidative enzymes like superoxide dismutase (SOD), catalase (CAT), guaiacol peroxidase (POD), glutathione reductase (GR), glutathione-S-transferase (GST) and dehydroascorbate reductase (DHAR). We further noticed that ALA upregulated the expression of SOD (7.30 folds), POD (6.11 folds), CAT (3.52 folds), DHAR (6.42 folds), GR (6.04 folds), and GST (5.58 folds) under the Pb stress. However, RBOH1 (gene involved in ROS generation) and CHLASE (chlorophyllase) expressions were reduced in ALA treated seedlings grown under Pb stress (RBOH1 expression decreased to 3.44 from 6.50 fold and CHLASE expression decreased to 2.97 from 5.58 fold). Phenolic contents were increased in the presence of ALA and expression of genes like CHS (chalcone synthase; 7.50 fold) and PAL (phenylalanine ammonia lyase; 4.77 fold) was also stimulated by ALA under Pb stress. Furthermore, contents of the Krebs cycle metabolites (fumarate, succinate, malate and citrate) were also enhanced accompanied by upregulated expression of genes like CS (citrate Synthase; 8.13 fold), SUCLG1 (succinyl CoA ligase 1; 7.40 fold), SDH (succinate dehydrogenase; 5.10 fold) and FH (fumarate hydratase; 5.65 fold). In conclusion, current investigation revealed that ALA attenuated Pb toxicity by modulating the transcription patterns of key enzymes involved in plant defense system.

ZnO nanoparticles induce cell wall remodeling and modify ROS/ RNS signalling in roots of Brassica seedlings.

Cell wall-associated defence against zinc oxide nanoparticles (ZnO NPs) as well as nitro-oxidative signalling and its consequences in plants are poorly examined. Therefore, this study compares the effect of chemically synthetized ZnO NPs (~45 nm, 25 or 100 mg/L) on Brassica napus and Brassica juncea seedlings. The effects on root biomass and viability suggest that B. napus is more tolerant to ZnO NP exposure relative to B. juncea. This may be due to the lack of Zn ion accumulation in the roots, which is related to the increase in the amount of lignin, suberin, pectin and in peroxidase activity in the roots of B. napus. TEM results indicate that root cell walls of 25 mg/L ZnO NP-treated B. napus may bind Zn ions. Additionally, callose accumulation possibly contribute to root shortening in both Brassica species as the effect of 100 mg/L ZnO NPs. Further results suggest that in the roots of the relatively sensitive B. juncea the levels of superoxide radical, hydrogen peroxide, hydrogen sulfide, nitric oxide, peroxinitrite and S-nitrosoglutathione increased as the effect of high ZnO NP concentration meaning that ZnO NP intensifies nitro-oxidative signalling. In B. napus; however, reactive oxygen species signalling was intensified, but reactive nitrogen species signalling wasn't activated by ZnO NPs. Collectively, these results indicate that ZnO NPs induce cell wall remodeling which may be associated with ZnO NP tolerance. Furthermore, plant tolerance against ZnO NPs is associated rather with nitrosative signalling than oxidative modifications.

Melatonin modulates photosynthesis, redox status, and elemental composition to promote growth of Brassica juncea-a dose-dependent effect.

Melatonin (MEL) is an antioxidant molecule, present throughout plant kingdom, animals, and microbes. It is a well-known free radical scavenger and modulates growth and development in plants against various abiotic and biotic stresses. The present study was done to investigate the role of MEL as a foliar spray on the morphological, physiological, and biochemical parameters in Brassica juncea cv. Varuna. Five different doses (10, 20, 30, 40, or 50 µM) of MEL were applied as foliar spray to the leaf of plant at 25 days after sowing (DAS) and continued up to 30 DAS once in a day. The plants were sampled at 30, 45, and 60 DAS to assess various parameters. The present results indicate that most of the parameters, i.e., growth, photosynthetic, nutrients, and enzyme activities increased in a concentration dependent manner. MEL application reduced the accumulation of reactive oxygen species (ROS) by enhancing the antioxidant enzyme activities. Microscopic examinations further revealed a significant increase in the size of the stomatal aperture in the presence of MEL. Out of the various concentrations tested, 40 µM of MEL proved best and can be used for further studies.

Phytoremediation of Cadmium Contaminated Soil Using Brassica juncea: Influence on PSII Activity, Leaf Gaseous Exchange, Carbohydrate Metabolism, Redox and Elemental Status.

Phytoremediation is an ecologically and economically feasible technique to remove heavy metal from soil. The aim of the study was to examine cadmium (Cd) toxicity and phytoremediation aptitude of Brassica juncea. In the present study, plants survived when exposed to different levels of Cd (0, 25, 50 and 100 mg/kg soil) and accumulated a large amount of Cd in its root and shoot. Translocation factor (TF) of Cd from root to shoot was > 1 at both 45 and 60-day stage of growth suggesting that B. juncea is a hyperaccumulator and strong candidate for phytoextraction of Cd. Alongside, Cd impaired photolysis of water, PSII activity, nutrient uptake, photosynthesis and sugar accumulation in the plant. Cd-generated oxidative stress restricts the growth of B. juncea. The toxic effect of Cd was more pronounced at 45-day stage of growth signifying the drifting of plant towards acquirement of exclusion strategy.

Postponement growth and antioxidative response of Brassica nigra on CuO and ZnO nanoparticles exposure under soil conditions.

Due to unique physiochemical properties, nanoparticles (NPs) have acquired substantial attention in the field of research. However, threats of ecotoxicity and phytotoxicity have limited their biological applications. In this study in vivo experiments were performed to determine the effect of CuO (12.5, 25 and 50 mg/kg) and ZnO (200, 400 and 600 mg/kg) NPs on growth, and antioxidant activities of Brassica nigra. The results showed that CuO NPs did not affect the seed germination while presence of ZnO NPs in the soil generated an inhibitory effect. Both CuO and ZnO NPs positively influenced the growth of stem and other physiological parameters i.e. stem height increased (23%) at 50 mg/kg CuO while root length decreased (up to 44%) with an increase in the concentration of NPs. Phytochemical screening of apical, middle and basal leaves showed elevated phenolic and flavonoid contents in the range of 15.3-59 µg Gallic Acid Equivalent (GAE)/mg Dry Weight (DW) and 10-35 µg Querceitin Equivalent (QE)/mg DW, respectively, in NPs-treated plants. Antioxidant activity was higher in CuO NPs-treated plants as compared to ZnO and control plants. Results conclude that CuO and ZnO NPs at low concentrations can be exploited as nanofertilisers in agriculture fields.

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.

Genome-wide identification and functional analysis of ARF transcription factors in Brassica juncea var. tumida.

Auxin signalling is vital for plant growth and development, from embryogenesis to senescence. Recent studies have shown that auxin regulates biological processes by mediating gene expression through a family of functionally original DNA-binding auxin response factors, which exist in a large multi-gene family in plants. However, to date, no information has been available about characteristics of the ARF gene family in Brassica juncea var. tumida. In this study, 65 B. juncea genes that encode ARF proteins were identified in the B. juncea whole-genome, classified into three phylogenetical groups and found to be widely and randomly distributed in the A-and B-genome. Highly conserved proteins were also found within each ortholog based on gene structure and conserved motifs, as well as clustering level. Furthermore, promoter cis-element analysis of BjARFs demonstrated that these genes affect the levels of plant hormones, such as auxin, salicylic, gibberellin acid, MeJA, abscisic acid, and ethylene. Expression analysis showed that differentially expressed BjARF genes were detected during the seedling stage, tumor stem development and the flowering period of B. juncea. Interestingly, we found that BjARF2b_A, BjARF3b_A, BjARF6b_A, and BjARF17a_B were significantly expressed in tumor stem, and an exogenous auxin assay indicated that these genes were sensitive to auxin and IAA signaling. Moreover, eight of the nine BjARF10/16/17 genes and all of the BjARF6/8 genes were involved in post-transcriptional regulation, targeted by Bj-miR160 and Bj-miR167c, respectively. This analysis provides deeper insight of diversification for ARFs and will facilitate further dissection of ARF gene function in B. juncea.

Interplay of Calcium and Nitric Oxide in improvement of Growth and Arsenic-induced Toxicity in Mustard Seedlings.

In this study, Ca2+ mediated NO signalling was studied in response to metalloid (As) stress in Brassica seedlings. Arsenic toxicity strongly suppressed the growth (fresh weight, root and shoot length), photosynthetic pigments, Chl a fluorescence indices (Kinetic traits: Fv, Fm, Fv/Fo, Fm/Fo, ФPo or Fv/Fm, Ψo, ФEo, PIABS, Area and N and redox status (AsA/DHA and GSH/GSSG ratios) of the cell; whereas energy flux traits: ABS/RC, TRo/RC, ETo/RC and DIo/RC along with Fo, Fo/Fv, Fo/Fm, ФDo and Sm) were enhanced. Further, addition of EGTA (Ca2+ scavenger) and LaCl3 (plasma membrane Ca2+ channel blocker) to As + Ca; while c‒PTIO (NO scavenger) and L‒NAME (NO synthase inhibitor) to As + SNP treated seedlings, siezed recovery on above parameters caused due to Ca2+ and NO supplementation, respectively to As stressed seedlings thereby indicating their signalling behaviour. Further, to investigate the link between Ca2+ and NO, when c‒PTIO and L‒NAME individually as well as in combination were supplemented to As + Ca treated seedlings; a sharp inhibition in above mentioned traits was observed even in presence of Ca2+, thereby signifying that NO plays crucial role in Ca2+ mediated signalling. In addition, As accumulation, ROS and their indices, antioxidant system, NO accumulation and thiol compounds were also studied that showed varied results.

Photosynthetic activity and RAPD profile of polyethylene glycol treated B. juncea L. under nitric oxide and abscisic acid application.

The involvement of two extremely important signalling molecules, nitric oxide (NO) and abscisic acid (ABA) has been employed by plants to facilitate the adaptive/tolerate response during stressful conditions. However, the interactive role of exogenously applied NO and ABA is very less studied at physiological, biochemical and molecular levels. The present study therefore, evaluated the effects of individual and simultaneous addition of exogenous NO donor SNP (100µM) and ABA (10µM) on photosynthesis, Calvin-Benson cycle enzymes, S-assimilation enzymes, oxidative stress components, and genotoxicity in Brassica juncea cv. Varuna, exposed to polyethylene glycol (PEG)-induced drought stress. Results showed that a loss induced by PEG was significantly surpassed by the application of NO or/and ABA with PEG for chlorophyll content, net photosynthestic rate (Pn), internal CO2 concentration (Ci), stomatal conductance (gs), transpiration rate (Tr), maximum photosystem II (PSII) efficiency (Fv/Fm), actual PSII efficiency (ΦPSII), intrinsic PSII efficiency (Fv´/ Fm´), photochemical quenching (qP), non-photochemical quenching (NPQ), electron transport chain (ETC), ribulose 1,5-bisphosphate carboxylase/oxygenase (RuBisCo), glyceraldehyde-3-phosphate dehydrogenase (GapDH), phosphoribulokinase (PRK), ATP-sulfurylase (ATP-S), and serine acetyltransferase (SAT) activities. The genomic template stability (GTS) (measured as changes in RAPD profiles) was significantly affected and showed varying degrees of DNA polymorphism, highest in PEG and lowest in PEG + NO and PEG + NO + ABA. Furthermore, the changes in RAPD profiles showed consistent results when compared with various photosynthetic and oxidative parameters. Altogether, this study concluded that supplementation of individual NO and together with ABA was more effective than individual ABA in alleviating PEG-induced drought stress in B. juncea L. seedlings.

Exogenous brassinosteroids increase lead stress tolerance in seed germination and seedling growth of Brassica juncea L.

Lead (Pb) is a highly toxic heavy metal to plants, animals, and human beings. The use of growth regulators has reversed the effects of heavy metal stress on germination and early plant development. The aim of this study was to evaluate the effect of brassinosteroids on seed germination and seedling growth of Brassica juncea (L.) Czern. & Coss. under Pb stress conditions. Two forms of application of 24-epibrassinolide (EBL) were evaluated, application on seeds in pre-soaking and on germination paper, using EBL concentrations of 0, 10-10, 10-8, and 10-6 M. Germination and seedling growth parameters were evaluated during the germination test. The activity of the enzymes superoxide dismutase, catalase, peroxidase, and ascorbate peroxidase were determined, as well as the lead content in the seeds and seedlings. The EBL applied at the 10-8 M concentration was the most effective in overcoming Pb stress in both forms of application. The antioxidant enzyme defense system was compromised by Pb exposure. However, 10-8 M EBL increased the activity of antioxidant enzymes such as catalase and peroxidase to overcome the toxic effects caused by Pb. In addition, EBL at the concentration of 10-8 M increased Pb content in seedlings without affecting seedling growth.

The endophytic bacterium Sphingomonas SaMR12 alleviates Cd stress in oilseed rape through regulation of the GSH-AsA cycle and antioxidative enzymes.

BACKGROUND: Microbes isolated from hyperaccumulating plants have been reported to be effective in achieving higher phytoextraction efficiency. The plant growth-promoting bacteria (PGPB) SaMR12 from the cadmium (Cd)/zinc hyperaccumulator Sedum alfredii Hance could promote the growth of a non-host plant, oilseed rape, under Cd stress. However, the effect of SaMR12 on Brasscia juncea antioxidative response under Cd exposure was still unclear. RESULTS: A hydroponic experiment was conducted to study the effects of Sphingomonas SaMR12 on its non-host plant Brassica juncea (L.) Czern. under four different Cd treatments. The results showed that SaMR12 could colonize and aggregate in the roots and then move to the shoots. SaMR12 inoculation promoted plant growth by up to 71% in aboveground biomass and 81% in root biomass over that of the non-inoculated plants. SaMR12-inoculated plants significantly enhanced root Cd accumulation in the 10 and 20 µM Cd treatments, with 1.72- and 0.86-fold increases, respectively, over that of the non-inoculated plants. SaMR12 inoculation not only decreased shoot hydrogen peroxide (H2O2) content by up to 38% and malondialdehyde (MDA) content by up to 60% but also reduced proline content by 7-30% in shoots and 17-32% in roots compared to the levels in non-inoculated plants. Additionally, SaMR12 inoculation promoted the activities of superoxide dismutase (SOD), peroxidase (POD), catalase (CAT), and ascorbate peroxidase (APX) and facilitated the relative gene expression levels of dehydroascorbate reductase (DHAR) and glutathione reductase (GR) involved in the glutathione (GSH)-ascorbic acid (AsA) cycle. CONCLUSIONS: The results demonstrated that, under Cd stress, SaMR12 inoculation could activate the antioxidative response of B. juncea by decreasing the concentrations of H2O2, MDA and proline, increasing the activities of antioxidative enzymes, and regulating the GSH-AsA cycle. These results provide a theoretical foundation for the potential application of hyperaccumulator endophytic bacteria as remediating agents to improve heavy metal tolerance within non-host plant species, which could further improve phytoextraction efficiency.

Silicon and plant growth promoting rhizobacteria differentially regulate AgNP-induced toxicity in Brassica juncea: Implication of nitric oxide.

An emerging stress of nanomaterials in soil and water is of great concern as it limits crop productivity and affects humans as well. Therefore, it is required to manage this problem. Silicon and plant growth promoting rhizobacteria has gained the engaging role in agriculture as (bio-)fertilizers. However, their role against silver nanoparticles (AgNPs) is still not known. Hence, present study was envisaged to investigate role of Si, PGPR and phytohormone indole acetic acid (IAA) in regulating AgNP stress in Brassica juncea seedlings. The study highlighted the impact of various treatments with respect to overproduction of reactive oxygen species, signaling molecule nitric oxide, oxidative markers like antioxidant enzymes and nonenzymatic components of ascorbate-glutathione pathway. Interestingly, silicon when present with AgNPs enhanced toxicity by reducing growth and mechanistic properties of B. juncea. Moreover, the results highlight the role of PGPR and IAA towards reduction in toxicity by promoting the plant growth under stressed conditions. Treatments AgNP + Si + PGPR/IAA were observed to significantly reduce the stress and enhance plant growth against treatment AgNPs alone. This reversal in toxicity by PGPR and IAA along with Si suggests the idea to formulate and utilize their combination as biofertilizers for eradicating the stress in near future.

Effects of endophytes inoculation on rhizosphere and endosphere microecology of Indian mustard (Brassica juncea) grown in vanadium-contaminated soil and its enhancement on phytoremediation.

We investigated the effects of endophytes inoculation on ecological factors such as root morphology, rhizosphere soil properties, heavy metal speciation, and rhizosphere and endophytic bacterial communities and their role on phytoremediation. Indian mustards were grown for two months in V-contaminated soil with three treatments (control, inoculation with Serratia PRE01 or Arthrobacter PRE05). Inoculation with PRE01 and PRE05 increased organic matter content by 6.94% and 4.6% respectively and significantly increased bioavailability of heavy metals in rhizosphere soils. Despite the endophyte inocula failed to flourish as stable endophytes, they significantly affected the specific composition and diversity of endophytic bacterial communities in roots, with no significant effect on rhizosphere bacterial communities. The test strains could greatly increase plant growth promotion-related biomarkers in the endosphere, especially those associated with Pseudomonas and Microbacterium genera. PICRUSt analysis predicted high relative abundances of functional genes related to environmental information processing especially in the endophytic microbiota. More biomass production (12.0%-17.4%) and total metals uptake (24.2%-32.0%) were acquired in inoculated treatments. We conclude that endophyte PRE01 or PRE05 inoculation could effectively enhance phytoremediation of V-contaminated soil by improving the rhizosphere and endosphere microecology without causing any ecological damage.