Citric acid assisted phytoextraction of nickle from soil helps to tolerate oxidative stress and expression profile of NRAMP genes in sunflower at different growth stages.

Introduction: Soil polluted with Nickel (Ni) adversely affects sunflower growth resulting in reduced yield. Counterbalancing Ni toxicity requires complex molecular, biochemical, and physiological mechanisms at the cellular, tissue, and whole plant levels, which might improve crop productivity. One of the primary adaptations to tolerate Ni toxicity is the enhanced production of antioxidant enzymes and the elevated expression of Ni responsive genes. Methods: In this study, biochemical parameters, production of ROS, antioxidants regulation, and expression of NRAMP metal transporter genes were studied under Ni stress in sunflower. There were four soil Ni treatments (0, 50, 100, and 200 mg kg-1 soil), while citric acid (CA, 5 mM kg-1 soil) was applied on the 28th and 58th days of plant growth. The samples for all analyses were obtained on the 30th and 60th day of plant growth, respectively. Results and discussion: The results indicated that the concentrations of Ni in roots and shoots were increased with increasing concentrations of Ni at both time intervals. Proline contents, ascorbic acid, protein, and total phenolics were reduced under Ni-stress, but with the application of CA, improvement was witnessed in their contents. The levels of malondialdehyde and hydrogen peroxide were enhanced with the increasing concentration of Ni, and after applying CA, they were reduced. The contents of antioxidants, i.e., catalase, peroxidase, superoxide dismutase, ascorbate peroxidase, dehydroascorbate reductase, and glutathione reductase, were increased at 50 ppm Ni concentration and decreased at higher concentrations of Ni. The application of CA significantly improved antioxidants at all concentrations of Ni. The enhanced expression of NRAMP1 (4, 51 and 81 folds) and NRAMP3 (1.05, 4 and 6 folds) was found at 50, 100 and 200ppm Ni-stress, respectively in 30 days old plants and the same pattern of expression was recorded in 60 days old plants. CA further enhanced the expression at both developmental stages. Conclusion: In conclusion, CA enhances Ni phytoextraction efficiency as well as protect plant against oxidative stress caused by Ni in sunflower.

Associative effects of lignin-derived biochar and arbuscular mycorrhizal fungi applied to soil polluted from Pb-acid batteries effluents on barley grain safety.

While disobeying environmental regulations of Pakistan, several Pb-acid batteries recycling and repairing units discharge their effluents into water canals that irrigate arable fields. Resultantly, serious ecological risks, as well as human health hazards through consumption of edible crops grown on such Pb-polluted soils have been reported. In this experiment, we observed associative effects of amending a soil polluted from Pb-acid batteries effluents (SPB) with arbuscular mycorrhizal fungi (AMF) and lignin-derived biochar (LBC) on barley grain safety to human health. The SPB was treated with AMF inoculum (a consortium of four AMF species), lignin (LN), and LBC, as sole treatments and AMF inoculum with LN and LBC. Barley parameters involving Pb distribution in grain and other parts, grain biochemistry, and nutrition were assessed. Likewise, Pb bioavailability in SPB, AMF root colonization, soil enzymes, microbial biomass carbon (MBC), and AMF produced total glomalin related soil protein (TGSP) were also scoped. Additionally, human renal cells (HEK 293) cytotoxicity test was performed by opting barley grain-related Pb concentrations. Results show that LBC + AMF significantly reduced grain Pb concentrations below the critical limit [4.67 mg kg-1 (WHO/FAO standard)], AMF colonization, MBC, soil enzymology, and TGSP, compared to control. Likewise, rest barley parameters were also improved in this treatment. Contrary to other treatments, grain produced on LBC + AMF did not result in (a) cell apoptosis, (b) cell distortion and (c) cohesion loss. Immobilization of Pb in SPB was due to the dilution effect of Pb adsorption on LBC, AMF mycelium and TGSP which resulted in a significant drop of grain Pb concentrations below the critical limit and ultimately no harm to HEK 293 cells. Our findings endorse that grain produced at LBC + AMF treatment are safer for human consumption and will not pose health risks. The LBC + AMF application can remediate SPB for safer cereal production.

Promoting the productivity and quality of brinjal aligned with heavy metals immobilization in a wastewater irrigated heavy metal polluted soil with biochar and chitosan.

Depleting aquifers, lack of planning and low socioeconomic status of Pakistani farmers have led them to use wastewater (WW) for irrigating their crops causing food contamination with heavy metals and ultimately negative effects on human health. This study evaluates the effects of chitosan (CH) and biochar (BC) on growth and nutritional quality of brinjal plant together with in situ immobilization of heavy metals in a soil polluted with heavy metals due to irrigation with wastewater (SPHIW) and further irrigated with the same WW. Both CH and BC were applied at three different rates i.e. low rate [(LR), BC0.5%, CH0.5% and BC0.25%+CH0.25%], medium rate [(MR), BC1%, CH1% and BC0.5%+CH0.5%] and high rate [(HR), BC1.5%, CH1.5% and BC0.75%+CH0.75%]. Result revealed that brinjal growth, antioxidant enzymes, and fruit nutritional quality significantly improved from LR to HR for each amendment, relative to control. However, these results were more prominent with BC alone and BC+CH, compared with CH alone at each rate. Similarly, with few exceptions, significant reduction in Ni, Cd, Co, Cr and Pb concentrations in the root, shoot and fruit were found in sole CH treatment both at LR and MR but in both CH and BC+CH treatments at HR, relative to control. Interestingly, the concentrations of Fe in the roots, shoots and fruit were more pronounced at BC treatments relative to CH and BC+CH treatments at each rate, compared to control. Overall, the BC+CH treatment at HR was the most effective treatment for in situ immobilization of heavy metals in SPHIW and further irrigated with the same WW, compared to rest of the treatments. This study indicates that BC0.75%+CH0.75% treatment can be used to reduce mobility and bioavailability of heavy metals in SPHIW and facilitates plant growth by improving the antioxidant system. However, the feasibility of BC0.75%+CH0.75% treatment should also be tested at the field scale.

Improvement in productivity, nutritional quality, and antioxidative defense mechanisms of sunflower (Helianthus annuus L.) and maize (Zea mays L.) in nickel contaminated soil amended with different biochar and zeolite ratios.

Nickel (Ni) contaminated soils pose a potential ecological risk to the environment, soil health, and quality of food produced on them. We hypothesized that application of miscanthus biochar (BC) and cationic zeolite (ZE) at various proportions into a Ni contaminated soil can efficiently immobilize Ni and reduce its bioavailability to sunflower (Helianthus annuus L.) and maize (Zea mays L.). An electroplating effluent contaminated soil was amended with BC and ZE, as sole treatments (2% w/w) and their combinations of various ratios (BC, ZE, BC25%ZE75%, BC50%ZE50% and BC75%ZE25%) for immobilization of Ni in the soil. Furthermore, the associated effects of these treatments on residual and DTPA-extractable Ni from the soil; concentrations of Ni in shoots, roots, and grain; growth, physiology, biochemistry and the antioxidant defence mechanisms of sunflower and maize were investigated. Results revealed that BC50%ZE50% treatment efficiently reduced DTPA-extractable Ni in the soil, Ni concentrations in shoots, roots, and grain, while improved selective parameters of both plants. Interestingly, the BC75%ZE25% treatment significantly improved the biomass, grain yield, physiology, biochemistry and antioxidant defense machinery, while decreased Ni oxidative stress in both sunflower and maize, compared to rest of the treatments. The results demonstrate that the BC50%ZE50% treatment can efficiently reduce Ni concentrations in the roots, shoots and grain of both sunflower and maize whereas, an improvement in biomass, grain yield, physiological, biochemical, and antioxidant defense machinery of both crops can only be achieved with the application of BC75%ZE25% treatment in a Ni contaminated soil.

Potential of miscanthus biochar to improve sandy soil health, in situ nickel immobilization in soil and nutritional quality of spinach.

The complex interaction of biochar (BC) with soil health reflecting properties, the feedstock used to prepare BC and application rate of BC in sandy soil is still a question for the researchers. An incubation study was conducted where nine different sorts of BC, each prepared from the different feedstock, were applied at 2% rate to evaluate their relative suitability to improve sandy soil health. Results revealed that BC prepared from miscanthus (MIB) significantly increased soil medium and fine pores, available water content (AWC), electrical conductivity (EC), and cation exchange capacity (CEC) while decreased soil wide pores, pH, bulk density (BD) and particle density (PD) compared to the rest sorts of BC. Later, spinach was grown in pots containing same soil but spiked with 50 ppm nickel (Ni) and amended with 1, 2, 3, 4 and 5% rates of MIB. The results showed a significant increment in spinach biomass, reduction in the concentrations of Ni in spinach tissues and DTPA-extractable Ni with the increasing rate of MIB till 3% and later, no significant changes with 4 and 5% rates thereafter. However, significant improvement in the activities of antioxidant enzymes, chemical and biochemical attributes of spinach were observed at 5% MIB when compared to lower rates. Similarly, post-harvest soil physicochemical and enzymatic parameters were also significantly (P < 0.05) improved with increasing rates of MIB. This study implies that application of MIB at 5% rate can improve the nutritional quality of spinach, sandy soil health and can reduce Ni concentrations in spinach tissues.

In situ immobilization of Cd by organic amendments and their effect on antioxidant enzyme defense mechanism in mung bean (Vigna radiata L.) seedlings.

In situ immobilization of Cd is desirable due to the damaging effects of ex situ remediation techniques on soil. In this greenhouse study, the role of biochar (BC), chitosan (CH), and green waste (GW) was studied for in-situ Cd immobilization and alleviating Cd toxicity in mung bean seedlings. Amendments were applied at rates of 0.5% and 1% (w/w). The minimum mean value of Cd, in root, shoot, and soil (DTPA-Cd) (12.2, 4.7, and 0.7 mg kg-1, respectively), occurred in the Cd + 1% CH treatment compared to all Cd amended treatments. Shoot dry weight (21%) increased significantly in Cd + 1% BC amended soil compared to the control. Reactive oxygen species were affected significantly, with the lowest increased value of hydrogen peroxide (4%) in the Cd + 1% CH treatment while the minimum increase in the value of superoxide (O2•-) occurred in the Cd + 1% BC soil compared to the control. Malondialdehyde (20%) increased lowest with Cd + 1% CH treatment. Protein, ascorbate (AsA) contents, and catalase (CAT) activity increased the most (3, 2, and 15%, respectively) in the Cd + 1% BC treatment while dehydroascorbate reductase (DHAR) and superoxide dismutase (SOD) activity increased the most (9 and 234%, respectively) in the Cd + 1% CH soil compared to the control. Glutathione reductase (GR), ascorbate peroxidase (APX), and glutathione peroxidase (GPX), activity were reduced the most in the Cd + 1.0% BC treatment while dehydroascorbate (DHA) and glutathione S-transferase (GST) activity decreased the most in the Cd + 1% CH soil. Overall, in situ immobilization by amendments improved growth and antioxidant defense mechanisms of mung bean seedlings and was reflected by tolerance to Cd-toxicity.

Improved quinoa growth, physiological response, and seed nutritional quality in three soils having different stresses by the application of acidified biochar and compost.

Quinoa (Chenopodium quinoa Willd.) is a traditional Andean agronomical resilient seed crop having immense significance in terms of high nutritional qualities and its tolerance against various abiotic stresses. However, finite work has been executed to evaluate the growth, physiological, chemical, biochemical, antioxidant properties, and mineral nutrients bioavailability of quinoa under abiotic stresses. Depending on the consistency in the stability of pH, intended rate of S was selected from four rates (0.1, 0.2, 0.3, 0.4 and 0.5% S) for the acidification of biochar and compost in the presence of Thiobacillus thiooxidans by pH value of 4. All three soils were amended with 1% (w/w) acidified biochar (BCA) and compost (COA). Results revealed that selective plant growth, yield, physiological, chemical and biochemical improved significantly by the application of BCA in all stressed soils. Antioxidants in quinoa fresh leaves increased in the order of control > COA > BCA, while reactive oxygen species decreased in the order of control < COA < BCA. A significant reduction in anti-nutrients (phytate and polyphenols) was observed in all stressed soils with the application of BCA. Moreover, incorporation of COA and BCA reduced the pH of rhizosphere soil by 0.4-1.6 units in all stressed soils, while only BCA in bulk soil decreased pH significantly by 0.3 units. These results demonstrate that BCA was more effective than COA to enhance the bioavailability, translocation of essential nutrients from the soil to plant and their enhanced bioavailability in the seed.

Cost-effective enhanced iron bioavailability in rice grain grown on calcareous soil by sulfur mediation and its effect on heavy metals mineralization.

Calcareous soil, high pH, and low organic matter are the major factors that limit iron (Fe) availability to rice crop. The present study was planned with the aim to biofortified rice grain with Fe, by integrated use of chemical and organic amendments in pH-manipulated calcareous soil. The soil pH was reduced (pHL2) by using elemental sulfur (S) at the rate of 0.25 % (w/w). The organic amendments, biochar (BC) and poultry manure (PM) [1 % (w/w)], along with ferrous sulfate at the rate of 7.5 mg kg-1 soil were used. The incorporation of Fe with BC in soil at pHL2 significantly improved plant biomass, photosynthetic rate, and paddy yield up to 99, 97, and 36 %, respectively, compared to control. A significant increase in grain Fe (190 %), protein (58 %), and ferritin (400 %) contents was observed while anti-nutrients, i.e., polyphenols (37 %) and phytate (21 %) were significantly decreased by the addition of Fe and BC in soil at pHL2 relative to control. Among the organic amendments, PM significantly increased Cd, Pb, Ni, and Cr concentrations in rice grain relative to control but their concentration values were below as compared to the toxic limits of hazard quotients and hazard index (HQ and HI). Hence, this study implies that Fe applied with BC in the soil at pHL2 can be considered as an effective strategy to augment Fe bioavailability and to reduce non-essential heavy metal accumulation in rice grain.

Effect of different amendments on rice (Oryza sativa L.) growth, yield, nutrient uptake and grain quality in Ni-contaminated soil.

Rice ( Oryza sativa L.) is one of the main staple food crops which is inherently low in micronutrients, especially iron (Fe), and can lead to severe Fe deficiency in populations having higher consumption of rice. Soils polluted with nickel (Ni) can cause toxicity to rice and decreased Fe uptake by rice plants. We investigated the potential role of biochar (BC) and gravel sludge (GS), alone and in combination, for in situ immobilization of Ni in an industrially Ni-contaminated soil at original and sulfur-amended altered soil pH. Our further aim was to increase Fe bioavailability to rice plants by the exogenous application of ferrous sulfate to the Ni-immobilized soil. Application of the mixture of both amendments reduced grain Ni concentration, phytate, Phytate/Fe, Phyt/Zn molar ratios, and soil DTPA-extractable Ni. In addition, the amendment mixture increased 70 % Fe and 229 % ferritin concentrations in rice grains grown in the soil at original pH. The Fe and ferritin concentrations in S-treated soil was increased up to 113 and 383 % relative to control respectively. This enhanced Fe concentration and corresponding ferritin in rice grains can be attributed to Ni/Fe antagonism where Ni has been immobilized by GS and BC mixture. This proposed technique can be used to enhance growth, yield, and Fe biofortification in rice by reducing soil pH while in parallel in situ immobilizing Ni in polluted soil.

Iron biofortification of wheat grains through integrated use of organic and chemical fertilizers in pH affected calcareous soil.

Incidence of iron (Fe) deficiency in human populations is an emerging global challenge. This study was conducted to evaluate the potential of iron sulphate combined with biochar and poultry manure for Fe biofortification of wheat grains in pH affected calcareous soil. In first two incubation studies, rates of sulfur (S) and Fe combined with various organic amendments for lowering pH and Fe availability in calcareous soil were optimized. In pot experiment, best rate of Fe along with biochar (BC) and poultry manure (PM) was evaluated for Fe biofortification of wheat in normal and S treated low pH calcareous soil. Fe applied with BC provided fair increase in root-shoot biomass and photosynthesis up to 79, 53 and 67%, respectively in S treated low pH soil than control. Grain Fe and ferritin concentration was increased up to 1.4 and 1.2 fold, respectively while phytate and polyphenol was decreased 35 and 44%, respectively than control in treatment where Fe was applied with BC and S. In conclusion, combined use of Fe and BC could be an effective approach to improve growth and grain Fe biofortification of wheat in pH affected calcareous soil.