Cd induced generation of free radical species in Brassica juncea is regulated by supplementation of earthworms in the drilosphere.

The antioxidant defense system of Brassica juncea under Cd stress was examined on supplementation of earthworms in the rhizosphere at different concentrations of Cd (0.50, 0.75, 1.00 and 1.25 mM i.e. 56, 84, 112 and 140 mg kg-1 respectively). Seedlings were raised in small pots containing soil spiked with Cd and earthworms under controlled conditions for 15 days. Improved Cd accumulation, as well as enhanced plant dry weight and metal tolerance were observed following the addition of earthworms. Earthworm supplementation reduced reactive oxygen species (ROS) generation by 7.3% for hydrogen peroxide (H2O2), 7.1% for superoxide anion (O2-), and 8.4% for malondialdehyde (MDA) in plants treated with 1.25 mM (140 mg kg-1) Cd. Confocal microscopy revealed improved cell viability and reduced H2O2 content due to enhanced antioxidative activity. Activity and expression levels of genes coding for antioxidative enzymes (superoxide dismutase; SOD, catalase; CAT, guaicol peroxidase; POD, glutathione reductase; GR, and glutathione-S-transferase; GST) were higher in plants raised in soils inoculated with earthworms, with expression of SOD increasing by 58.8%, CAT by 75%, POD by 183%, GR by 106.6%, and GST by 11.8%. Moreover, plant pigment (chlorophyll a, chlorophyll b, total chlorophyll, and carotenoids) concentrations increased by 8%, 9.1%, 9.1%, and 7.7% respectively, in plants grown in soils supplemented with earthworms. The results of our study suggest that the addition of earthworms to soil increases antioxidative enzyme activities, gene expression in plants, and ROS inhibition, which enhances tolerance to Cd during the phytoextraction process.

Kaur P ，Bali S ，Sharma A ，Kohli SK ，Vig AP ，Bhardwaj R ，Thukral AK ，Abd Allah EF ，Wijaya L ，Alyemeni MN ，Ahmad P ... -  《-》

Insights into citric acid-induced cadmium tolerance and phytoremediation in Brassica juncea L.: Coordinated functions of metal chelation, antioxidant defense and glyoxalase systems.

Cadmium (Cd) is a serious environmental threat because it accumulates in plants from soil and is subsequently transported into the food cycle. Increased Cd uptake in plants disrupts plant metabolism and hampers crop growth and development. Therefore, remediation of Cd from soil and enhancing plant tolerance to metal toxicity is vital. In the present study, we investigated the function of different doses of citric acid (CA) on Cd toxicity in terms of metal accumulation and stress tolerance in mustard (Brassica juncea L.). Brassica juncea seedlings (12-day-old) were treated with Cd (0.5mMCd and 1.0mM CdCl2) alone and in combination with CA (0.5mM and 1.0mM) in a semi-hydroponic medium for three days. Cadmium accumulation in the roots and shoots of the mustard seedlings increased in a dose-dependent manner and was higher in the roots. Increasing the Cd concentration led to reduced growth, biomass, water status, and chlorophyll (chl) content resulting from increased oxidative damage (elevated malondialdehyde, MDA content; hydrogen peroxide, H2O2 level; superoxide, O2•- generation; lipoxygenase, LOX activity; and methylglyoxal, MG content) and downregulating of the major enzymes of the antioxidant defense and glyoxalase systems. Under Cd stress, both doses of CA improved the growth of the plants by enhancing leaf relative water content (RWC) and chl content; reducing oxidative damage; enhancing the pool of ascorbate (AsA) and glutathione (GSH) and the activities of the antioxidant enzymes (ascorbate peroxidase, APX; monodehydroascorbate reductase, MDHAR; dehydroascorbate reductase, DHAR; glutathione reductase, GR; glutathione peroxidase, GPX; superoxide dismutase, SOD; catalase, CAT); improving the performance of the glyoxalase system (glyoxalase I, Gly I and glyoxalase II, Gly II activity); and increasing the phytochelatin (PC) content. Exogenous CA also increased the root and shoot Cd content and Cd translocation from the roots to the shoots in a dose-dependent manner. Our findings suggest that CA plays a dual role in mustard seedlings by increasing phytoremediation and enhancing stress tolerance through upregulating the antioxidant defense and glyoxalase systems.

Mahmud JA ，Hasanuzzaman M ，Nahar K ，Bhuyan MHMB ，Fujita M ... -  《-》

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.

Singh R ，Kesavan AK ，Landi M ，Kaur S ，Thakur S ，Zheng B ，Bhardwaj R ，Sharma A ... -  《-》

Interaction of 24-epibrassinolide and salicylic acid regulates pigment contents, antioxidative defense responses, and gene expression in Brassica juncea L. seedlings under Pb stress.

Kohli SK ，Handa N ，Sharma A ，Gautam V ，Arora S ，Bhardwaj R ，Wijaya L ，Alyemeni MN ，Ahmad P ... -  《-》

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.

Yadav P ，Kaur R ，Kanwar MK ，Sharma A ，Verma V ，Sirhindi G ，Bhardwaj R ... -  《-》