Copper Oxide Nanoparticles Induced Growth and Physio-Biochemical Changes in Maize (Zea mays L.) in Saline Soil.

Research on nanoparticles (NPs) is gaining great attention in modulating abiotic stress tolerance and improving crop productivity. Therefore, this investigation was carried out to evaluate the effects of copper oxide nanoparticles (CuO-NPs) on growth and biochemical characteristics in two maize hybrids (YH-5427 and FH-1046) grown under normal conditions or subjected to saline stress. A pot-culture experiment was carried out in the Botanical Research Area of "the University of Lahore", Lahore, Pakistan, in a completely randomized design. At two phenological stages, both maize hybrids were irrigated with the same amount of distilled water or NaCl solution (EC = 5 dS m-1) and subjected or not to foliar treatment with a suspension of CuO-NPs. The salt stress significantly reduced the photosynthetic parameters (photosynthetic rate, transpiration, stomatal conductance), while the sodium content in the shoot and root increased. The foliar spray with CuO-NPs improved the growth and photosynthetic attributes, along with the N, P, K, Ca, and Mg content in the roots and shoots. However, the maize hybrid YH-5427 responded better than the other hybrid to the saline stress when sprayed with CuO-NPs. Overall, the findings of the current investigation demonstrated that CuO-NPs can help to reduce the adverse effects of salinity stress on maize plants by improving growth and physio-biochemical attributes.

Lead Toxicity-Mediated Growth and Metabolic Alterations at Early Seedling Stages of Maize (Zea mays L.).

To investigate the toxic effects of lead (Pb) on key metabolic activities essential for proper germination and seedling growth of maize seeds, experiments were carried out with different levels of Pb (0 to 120 mg of Pb L-1 as PbCl2) applied through growth medium to two maize hybrids H-3310S and H-6724. The research findings indicated that growth and metabolic activities were adversely affected by increased Pb contamination in growth medium; however, a slow increase in these parameters was recorded with increasing time from 0 to 120 h. Protease activity decreased with an increase in the level of Pb contamination but increased with time; consequently, a reduction in seed proteins and an increase in total free amino acids were observed with time. Similarly, α-amylase activity decreased with an increase in Pb concentration in growth medium while it increased with increasing time from 0 to 120 h; consequently, reducing and non-reducing sugars increased with time but decreased with exposure to lead. The roots of both maize hybrids had higher Pb contents than those of the shoot, which decreased the uptake of nitrogen, phosphorus, and potassium. All these nutrients are essential for optimal plant growth; therefore, the reduction in growth and biomass of maize seedlings could be due to Pb toxicity that altered metabolic processes, as sugar and amino acids are necessary for the synthesis of metabolic compounds, rapid cell division, and proper functioning of enzymes in the growing embryo, but all were dramatically reduced due to suppression of protease and α-amylase by toxicity of Pb. In general, hybrid H-3310S performed better in Pb-contaminated growth medium than H-6724.

Potential effect of luteolin, epiafzelechin, and albigenin on rats under cadmium-induced inflammatory insult: In silico and in vivo approach.

Introduction: Cadmium(Cd) an industrial poison present abundantly in the environment, causes human toxicity by an inflammatory process. Chronic exposure of cadmium can cause a number of molecular lesions that could be relevant to oncogenesis, through indirect or epigenetic mechanisms, potentially including abnormal activation of oncogenes and suppression of apoptosis by depletion of antioxidants. As induction of cyclooxygenase (COX)-2 is linked to inflammatory processes, use of luteolin, epiafzelechin, and albigenin alone or in different combinations may be used as anti-inflammatory therapeutic agents. Methods: We, herein, performed in silico experiments to check the binding affinity of phytochemicals and their therapeutic effect against COX-2 in cadmium administered rats. Wistar albino rats were given phytochemicals in different combinations to check their anti-inflammatory activities against cadmium intoxication. The level of alanine aminotransferases (ALT), 4-hydroxynonenal (4HNE), 8-hydroxy-2-deoxyguanosine (8-OHdG), tumor necrosis factor-alpha (TNF-α), isoprostanes (IsoP-2α), COX-2, and malondialdehyde (MDA) were estimated with their respective ELISA and spectrophotometric methods. Results: The generated results show that phytocompounds possessed good binding energy potential against COX-2, and common interactive behavior was observed in all docking studies. Moreover, the level of ALT, 4HNE, 8-OHdG, TNF-α, IsoP-2α, malondialdehyde, and COX-2 were significantly increased in rats with induced toxicity compared to the control group, whereas in combinational therapy of phytocompounds, the levels were significantly decreased in the group. Discussion: Taken together, luteolin, epiafzelechin, and albigenin can be used as anti-inflammatory therapeutic agents for future novel drug design, and thus it may have therapeutic importance against cadmium toxicity.

Determination and detoxification of aflatoxin and ochratoxin in maize from different regions of Pakistan.

The contamination of food commodities with mycotoxins could be a serious health threat to humans and animals. Therefore, identification, quantification and reduction of mycotoxins in food commodities, particularly of aflatoxins (AFs) and ochratoxin A (OTA) in grain foods, is essentially required to guarantee safe food. This study determined the levels of AFs and OTA in 135 maize grains samples belonging to eight salient maize varieties cultivated in Pakistan, and evaluated the usefulness of radiations and adsorbents to reduce their levels. High performance liquid chromatography (HPLC)-based method was validated for the determination of AFs and OTA in maize grains. The results showed that 69 and 61% samples were positive for AFs and OTA, respectively and 54 and 22% of the respective samples had AFs and OTA above the permissible limits set by Pakistan Standards and Quality Control Authority. The concentration of AFs, AFB1and OTA in grains ranged from 14.5 to 92.4, 1.02 to 2.46 and 1.41 to 53.9 µg kg-1, respectively. Among the varieties, Pearl had the highest level of total AFs and OTA, whereas YH-5427 had the highest AFB1 level. The lowest concentration of AFs and OTA was found in Malaka and 30Y87, respectively. The use of 15 kGy gamma irradiation for 24 h, sunlight-drying for 20 h and UV irradiation for 12 h almost completely degraded the mycotoxins. The microwave heating for 120 s resulted in 9-33% degradation of mycotoxins. Moreover, the treatment of grains' extract with activated charcoal (5% w/w) removed > 96% of total AFs and AFB1, and up to 43% of OTA. The use of bentonite at the same rate removed OTA, total AFs and AFB1 by 93, 73 and 92%, respectively. Thus, it is concluded that contamination of maize grains with mycotoxins was fairly high in the collected maize grain samples in Pakistan, and treatment with radiations and adsorbents can effectively reduce mycotoxins contamination level in maize grains.

Simultaneous Biofortification of Rice With Zinc, Iodine, Iron and Selenium Through Foliar Treatment of a Micronutrient Cocktail in Five Countries.

Widespread malnutrition of zinc (Zn), iodine (I), iron (Fe) and selenium (Se), known as hidden hunger, represents a predominant cause of several health complications in human populations where rice (Oryza sativa L.) is the major staple food. Therefore, increasing concentrations of these micronutrients in rice grain represents a sustainable solution to hidden hunger. This study aimed at enhancing concentration of Zn, I, Fe and Se in rice grains by agronomic biofortification. We evaluated effects of foliar application of Zn, I, Fe and Se on grain yield and grain concentration of these micronutrients in rice grown at 21 field sites during 2015 to 2017 in Brazil, China, India, Pakistan and Thailand. Experimental treatments were: (i) local control (LC); (ii) foliar Zn; (iii) foliar I; and (iv) foliar micronutrient cocktail (i.e., Zn + I + Fe + Se). Foliar-applied Zn, I, Fe or Se did not affect rice grain yield. However, brown rice Zn increased with foliar Zn and micronutrient cocktail treatments at all except three field sites. On average, brown rice Zn increased from 21.4 mg kg-1 to 28.1 mg kg-1 with the application of Zn alone and to 26.8 mg kg-1 with the micronutrient cocktail solution. Brown rice I showed particular enhancements and increased from 11 µg kg-1 to 204 µg kg-1 with the application of I alone and to 181 µg kg-1 with the cocktail. Grain Se also responded very positively to foliar spray of micronutrients and increased from 95 to 380 µg kg-1. By contrast, grain Fe was increased by the same cocktail spray at only two sites. There was no relationship between soil extractable concentrations of these micronutrients with their grain concentrations. The results demonstrate that irrespective of the rice cultivars used and the diverse soil conditions existing in five major rice-producing countries, the foliar application of the micronutrient cocktail solution was highly effective in increasing grain Zn, I and Se. Adoption of this agronomic practice in the target countries would contribute significantly to the daily micronutrient intake and alleviation of micronutrient malnutrition in human populations.

Nitric oxide regulates water status and associated enzymatic pathways to inhibit nutrients imbalance in maize (Zea mays L.) under drought stress.

Nitric oxide (NO) is a key signaling molecule that instigates significant changes in plant metabolic processes and promotes tolerance against various environmental stresses including drought. In this study, we focused on NO-mediated physiological mechanisms and enzymatic activities that influence the nutrient concentrations and yield in maize under drought stress. The drought-tolerant (NK-8711) and sensitive (P-1574) maize hybrids were sown in lysimeter tanks and two levels of water stress (well-watered at100% field capacity and drought stress at 60% field capacity) were applied at three-leaves stage of maize. Foliar treatment of sodium nitroprusside (SNP), the donor of NO was applied at the cob development stage. The results showed that the foliar spray of NO regulated water relations by increasing proline content and improved drought tolerance in water stressed maize plants. In addition, it stimulated the activity of antioxidative enzymes which reduced the production of free radicals and lipid peroxidation. The activities of nitrate assimilation enzymes were considerably increased by NO spray which, in turn, increased nutrient accumulation and yield in maize under water deficit conditions. These results acknowledge the importance of NO as a stress-signaling molecule that positively regulates defense mechanisms in maize to withstand water-limited conditions.

Unusual pattern of aflatoxins and ochratoxin in commercially grown maize varieties of Pakistan.

The contamination of food with mycotoxins could be a major health risk. Identification and quantification of mycotoxins in maize are essentially required to guarantee food safety. Seven commonly grown maize cultivars were sown at semi-arid conditions in Pakistan. After harvesting plants, total aflatoxin (AFs), aflatoxins B1 (AFB1), B2 (AFB2), G1 (AFG1), G2 (AFG2), and ochratoxin-A (OTA) contents were determined in maize flour using high performance liquid chromatography (HPLC). Results showed an unusual pattern of AFs in maize with a higher concentration of AFG1 in all maize varieties. The AFB1 was not found in most of the samples, however, all maize grains contained AFs above the maximum tolerable limit (20 µg/kg). The highest concentration of AFs was recorded in Ayub-1/26 maize (362.8 µg/kg). Ochratoxin-A (OTA) was detected in 71% maize samples ranged from 2.14 to 214 µg/kg. The highest OTA and microbial load were recorded in Ayub-2/27 grains that exceeded the FDA limit (50 µg/kg). These results indicated the higher prevalence of total AFs, AFG1, and OTA at the harvest stage of maize that could pose significant health hazards to humans and livestock.

Assessment of Cucumber Genotypes for Salt Tolerance Based on Germination and Physiological Indices.

Soil salinity is one of the primary problem for agricultural crops which causes a great loss in crop production in Pakistan and worldwide. Various approaches have been implemented to overcome salinity problem. Assembly of crops for the enhancement of salt tolerance is a good strategy to achieve cost-effective yields. Cucumber is considered as one of the leading vegetable crop around the world for the nourishment of human being as source of nutrients, minerals, and vitamins. Screening of 12 cucumber genotypes using some physiological indices, that is, seedling germination stress tolerance index, plant height stress tolerance index, root length stress tolerance index, shoot and root dry weight stress tolerance index, and shoot and root fresh weight stress tolerance index were performed for the identification of salt tolerance. Using the above characteristics genotypes, Valley and HC-999 were categorized as tolerant, Safaa and Debra as medium tolerant, while Thamin-II identified as medium sensitive and NSC-CM1 and Akbar are classified as sensitive genotypes of cucumber. According to the current study findings, the screened cucumber genotypes for salinity tolerance can also be suggested to farmers for the improved production and yield of crop at saline soil.

Ni tolerance and its distinguished amelioration by chelating agents is reflected in root radius of B. napus cultivars.

The negative effect of excess nickel (Ni) on plants is well investigated but there is only little information on its influence on root anatomy and a possible amelioration by chelating agents. In this study, we utilized light microscopy to observe anatomical changes in canola (Brassica napus) roots and investigated the element content by X-ray microanalysis. Ni-tolerant (Con-II) and Ni-sensitive cultivars (Oscar) were selected for this purpose. The plants were treated with 30 ppm NiSO4. Then, citric acid and ethylene-diamine-tetra-acetic acid (EDTA) (alone or in combination) were applied to observe the influence of chelating agents in metal stress amelioration. Ni treatment led to significant swelling of the roots in the Con-II variety as compared to the cultivar Oscar. Application of EDTA reduced the root radius of Con-II plants and this effect for Ni tolerance is discussed. According to X-ray microanalyses, Ni ions were more dispersed in the sensitive cultivar as indicated by metal adsorption to the cell wall. We investigate the hypothesis that an enhanced capacity of binding metals to the cell wall allows the plants to tolerate more heavy metals.

Exogenously applied zinc and copper mitigate salinity effect in maize (Zea mays L.) by improving key physiological and biochemical attributes.

Zinc or copper deficiency and salinity are known soil problems and often occur simultaneously in agriculture soils. Plants undergo various changes in physiological and biochemical processes to respond to high salt in the growing medium. There is lack of information on the relation of exogenous application of Zn and Cu with important salinity tolerance mechanisms in plants. Therefore, the present study was conducted to determine the effect of foliar Zn and Cu on two maize cultivars (salt-tolerant cv. Yousafwala Hybrid and salt-sensitive cv. Hybrid 1898). Salinity caused a significant reduction in water and turgor potentials, stomatal conductance, and transpiration and photosynthetic rate, while increase in glycine betaine, proline, total soluble sugars, and total free amino acids was evident in plants under saline regimes. Furthermore, there was significant decline in P, N, Ca, K, Mn, Fe, Zn, and Cu and increase in Na and Cl contents in plants fed with NaCl salinity. Nitrate reductase activity was lower in salt-stressed plants. However, foliar application of Zn and Cu circumvented salinity effect on water relations, photosynthesis, and nutrition and this was attributed to the better antioxidant system and enhanced accumulation of glycine betaine, proline, total free amino acids, and sugars. The results of the present study suggested that Zn application was superior to Cu for mediating plant defense responses under salinity.

Pb-induced phytotoxicity in para grass (Brachiaria mutica) and Castorbean (Ricinus communis L.): Antioxidant and ultrastructural studies.

Hydroponics experiment was conducted to investigate the effects of different levels of Pb on Para Grass (Brachiaria mutica) and Castorbean (Ricinus communis L). Generally, Para Grass exhibited higher tolerance to excessive concentrations of Pb in nutrient solution, whereas a consistent decline was observed in growth of Castorbean plants exposed to similar Pb levels. Malondialdehyde (MDA) and H2O2 contents exhibited contrasting results with a general decrease in Para Grass and a linear increase in case of Castorbean. In both species a decrease was noticed in the activities of superoxide dismutase (SOD) and guaiacol peroxidase (G-POD) while catalase (CAT) activity was significantly increased. Ultrastructural studies revealed increased starch grains and adversely affected thylakoid membranes in chloroplasts of leaf cells of plants treated with 500 µM Pb. Photosynthetic parameters such as CO2 assimilation rate, stomatal conductance (gs) and transpiration rate (E) decreased in both plant species under different levels of Pb. Maximum concentrations of Pb in shoots of Para Grass and Castorbean were 1.29 and 0.352 g kg-1, respectively while in roots maximum values were 8.88 and 49.86 g kg-1, respectively. The high concentrations of Pb (about 5%) in the roots of Castorbean plants suggest its possible role in the phytoremediation/rhizofiltration of Pb contaminated water.

Understanding brassinosteroid-regulated mechanisms to improve stress tolerance in plants: a critical review.

Brassinosteroids (BRs) are steroidal plant hormones involved in regulation of physiological and molecular processes to ameliorate various biotic and abiotic stresses. Exogenous application of BRs to improve stress tolerance in plants has recently become a high research priority. Several studies have revealed the involvement of these steroidal hormones in upregulation of stress-related defense genes and their cross talk with other metabolic pathways. This is likely to stimulate research on many unanswered questions regarding their role in enhancing the ability of plants to tolerate adverse environmental conditions. Thus, this review appraises new insights on mechanisms mediating BR-regulated changes in plants, focused mainly on their involvement in regulation of physiological and molecular mechanisms under stress conditions. Herein, examples of BR-stimulated modulation of antioxidant defense system and upregulation of transcription factors in plants exposed to various biotic (bacterial, viral, and fungal attack) and abiotic stresses (drought, salinity, heat, low temperature, and heavy metal stress) are discussed. Based on these insights, future research in the current direction can be helpful to increase our understanding of BR-mediated complex and interrelated processes under stress conditions.

Organic chelants-mediated enhanced lead (Pb) uptake and accumulation is associated with higher activity of enzymatic antioxidants in spinach (Spinacea oleracea L.).

The spinach was tested in the present studies for its phytoextraction potential. Furthermore, the study assessed whether organic chelants could reduce oxidative stress, and thus enhance growth of spinach plants under 2.42 and 4.83mM Pb regimes. Different organic chelates viz. ethylenediamine tetra acetic acid, (EDTA), citric acid (CA), oxalic acid (OA), tartaric acid (TA) and malic acid (MA) were applied separately in addition to control (without chelating agents) under different Pb regimes. The low (2.42mM) Pb regime increased biological yield (kgha(-1)). All the chelates except OA increased biological yield under low Pb regime. In contrast, TA caused less decrease in biomass under high (4.83mM) Pb regime. The chelate-assisted rise in the antioxidant activities substantially contributed to reactive oxygen species (ROS) neutralization. Of the chelates, TA was the most effective in improving Pb uptake and its root to shoot translocation. Overall, the chelate-assisted buildup of Pb in the spinach did not exhibit inhibitory effects on the plant growth possibly due to their potential to decrease Pb-induced oxidative damage. The results elaborated the potential of TA in increasing root to shoot translocation of Pb, biomass, and thus suggested its use for phytoextraction of Pb using spinach in Pb contaminated environments.

Salinity and drought induced antioxidant responses in different cultivars of safflower (Carthamus tinctorius L.).

Six cultivars of safflower which were (PI-387820, PI-251978, PI-170274, PI-387821, PI-386174 and Thori-78) grown in net house of NIAB under salinity (10 ds m⁻¹) and drought (60% field capacity) conditions and compared to their oxidative damage and antioxidative responses. Plant samples (leaves) were collected for the determination of malonidialdehyde (MDA), antioxidative enzymes (catalase, ascorbate peroxidase, glutathione reductase, and peroxidase), proline, and photosynthetic pigments. Salinity and drought decreased the chlorophyll a and b contents but a decrease in chlorophyll a and b was less in safflower variety (THORI-78) which could be a useful marker for selecting a stress tolerant variety. Both stresses considerable increases the accumulation of proline in PI-251978, PI-170274, PI-387821, PI-386174 and THORI-78 varieties of safflower whereas the proline accumulation did not appear to be an essential part of the protection mechanism against salinity and drought in variety PI-387820. Enzyme activity measurements revealed that THORI-78 can tolerate salinity and drought stress well by increasing the activity of catalase and APX enzymes whereas variety PI- 386174 showed increased activity of glutathione reductase enzyme under salinity and drought and appear to be very crucial antioxidative defenses during intense stress conditions. The results indicate that the photosynthetic pigments, proline and activities of the enzymes are important mechanism for the stress tolerance in safflower plant and can be considered as genetic improvement for the plant in salinity and drought soil conditions.

Toxic effect of nickel (Ni) on growth and metabolism in germinating seeds of sunflower (Helianthus annuus L.).

To assess the toxic effect of nickel (Ni) on the growth and some key metabolic processes in sunflower, varying levels of Ni as Ni(NO(3))(2) up to 60 mg L(-1) were applied once to sunflower cultivars SF-187 and Hysun-33 at sowing time in sand culture. An increase in Ni in the growth medium adversely affected growth parameters, sugar concentration (both reducing and non-reducing), as well as the activities of α-amylase and protease. It also slowed down mobilization of stored proteins and amino acids in the germinating seeds. However, an increase in the activities of α-amylase and protease was observed over time from 24 to 120 h after sowing. Cultivar Hysun-33 showed better performance than SF-187 in the presence of excess Ni. Overall, Ni-induced reduction in germination of sunflower seed appeared to be due to disturbance in biochemical metabolism as the availability of sugars for the synthesis of metabolic energy as well as necessary amino acids for the synthesis of proteins and enzymes essential for the growing embryo are generally reduced due to suppression in α-amylase and protease activities.

Influence of lead on growth and nutrient accumulation in canola (Brassica napus L.) cultivars.

Canola (Brassica napus L.) is commonly used as a hyper-accumulator for phytoextraction of heavy metals from soil and water. Like many other heavy metals, lead (Pb) contaminates soil, water and air and thus it is a great problem. This study was conducted to investigate toxic effects of Pb on growth and nutrient uptake in four canola cultivars. Each of four cultivars of canola (Con-II, Con-III, Legend and Shiralee) was subjected to four levels of Pb (0, 30, 60 and 90 mg Pb kg(-1) of soil) from lead chloride [PbCl2]. Due to Pb toxicity, plant growth was adversely affected and relatively a severe reduction in root biomass (45.7%) was recorded. The Pb accumulation increased both in shoot and root, the highest being in root. The uptake of different nutrients, i.e., N, P, K, Ca, Mg, Zn, Cu and Mn was reduced (38.4, 32.8, 33.1, 49.6, 7.78, 52.0, 42.6 and 45.9%, respectively) in the shoots and that of N, Fe, Zn, and Cu in the roots (48.5, 33.2, 24.3 and 44.8%, respectively) of all canola cultivars. The root K, P, Zn and Mn and shoot P, Mg and Fe contents were less affected, the concentration of Pb, Ca and Mg in roots of all cultivars. Among canola cultivars Con-II and Con-III performed better than Legend and Shiralee in terms of growth (26.03%) and nutrient accumulation. Overall, plant growth and nutrient accumulation in the canola cultivars was hampered due to the presence of Pb.

Therapeutic effects of ciprofloxacin on the pharmacokinetics of carbamazepine in healthy adult male volunteers.

Carbamazepine is a (CYP1A2 and CYP3A4 enzyme inducer) medicine which is used by epileptic patients for a long time. During the course of therapy, patients are generally caught by other diseases like urinary tract infections, upper respiratory tract infection, skin and soft tissue infection etc. To cure them, physicians commonly prescribe fluoroquinolones like Ciprofloxacin (CYP1A2 inhibitor) along with Carbamazepine (CBZ). Interactions may result without recognition which may lead to unforeseen toxicity, untoward effects or even therapeutic failure. Therefore, studies were conducted to investigate the effect of Ciprofloxacin on the pharmacokinetics of carbamazepine in healthy adult male volunteers. The main objective of this study was to generate new knowledge regarding CBZ and ciprofloxacin interaction for physicians and research workers dealing with these medicines. Eight healthy adult male volunteers were selected to assess the effect of ciprofloxacin on the pharmacokinetics of carbamazepine. After overnight fast the selected male volunteers were given CBZ orally. Blood samples were drawn at different time intervals after medication. Then the same volunteers were given CBZ along with ciprofloxacin. Blood samples were again drawn at the same time intervals as done previously. Plasma was separated from the blood samples. Concentration of CBZ in the plasma samples was determined by using HPLC technique. Results of the present study indicated that ciprofloxacin significantly increased the plasma concentration of CBZ when given concurrently to the healthy adult male volunteers. Ciprofloxacin increased Cmax, AUC and t½ while it decreased the CL and Vd of CBZ when administered concurrently to the adult volunteers. Change in pharmacokinetic parameters was due to slow metabolism or elimination of CBZ when given concurrently with ciprofloxacin to the adult volunteers. This is probably due to the inhibition of CYP3A4 isoenzyme by ciprofloxacin which is responsible for metabolism of CBZ. Ciprofloxacin increased the plasma concentration of CBZ so dose adjustment as well as drug monitoring of CBZ is required when both the drugs are given concurrently. The knowledge regarding interaction between ciprofloxacin and CBZ would be helpful for the pharmaceutical industries, physicians and a blessing for the patients.