ZnSO4 Treatment Research Articles

Cloning and expression analysis of metallothionein family genes in response to heavy metal stress in sugarcane (<italic>Saccharum officinarum</italic> L.)

Metallothioneins (MTs) are cysteine-rich, low-molecular-weight proteins. Plant MTs play important roles in detoxification and cellular redox regulation. In this study, hydroponic experiments of sugarcane ( Saccharum officinarum L. cv. Badila) were carried out to study the effects of CdCl2, ZnSO4, and CuCl2 treatments on plantlet growth. And then three kinds of heavy metal content in both shoots and roots of sugarcane were detected, showing an enrichment and tolerance ability to Cd2+, Zn2+, and Cu2+ in Badila seedlings. Three metallothionein genes, termed as ScMT 1 (accession number: KJ504373), ScMT 2-1-5 (accession number: MH191346), and ScMT 3 (accession number: KJ5043704), were isolated from Badila. ScMT 1 contained an open reading frame (ORF) of 228 bp encoding 75 amino acids residues. The first (52 bp) and second (176 bp) extrons of ScMT 1 were separated by an intron (283 bp). ScMT 2-1-5 contained an ORF of 246 bp encoding 81 amino acids residues. The first (64 bp), second (87 bp), and third (101 bp) extrons of ScMT2-1-5 were separated by two introns (483 bp and 853 bp). ScMT 3 contained an ORF of 198 bp encoding 65 amino acids residues. The first (50 bp) and second (148 bp) extrons of ScMT 3 were separated by an intron (259 bp). The deduce protein ScMT1, ScMT2-1-5, and ScMT3 were categorized into the subfamily of plant type1, type2, and type3 MTs, respectively. The expression profiles of the three genes under different heavy-metal stresses were investigated by real-time quantitative PCR (qRT-PCR) analysis. When treated with Cd2+, the expression of ScMT 2-1-5 was continuously and significantly up-regulated in both shoots and roots, while that of ScMT 1 showed a delayed up-regulation pattern. The expression of ScMT 3 showed a delayed up-regulation pattern in shoots and a trend of first raising and then suppressing in roots. The results suggested that ScMT 2-1-5 might play a more active role in response to Cd2+ in sugarcane while ScMT 3 might not, and ScMT 1 is involved in the molecular responses of Cd2+ stress at the later stage. When treated with Cu2+, the expression of ScMT 1 was continuously and significantly up-regulated, and that of ScMT 2-1-5 and ScMT 3 general up-regulated in shoots. As in the roots, the expression of both ScMT 1 and ScMT 2-1-5 showed a delayed up-regulation pattern which was significantly up-regulated at the later stage, while that of ScMT 3 significantly up-regulated at the early stage. It revealed that ScMT 1 cooperated with ScMT 2-1-5 and ScMT 3 is involved in the positive response to Cu2+ stress in roots, and might play a more positive role than ScMT 2-1-5 and ScMT 3, and the three genes are successively involved in the molecular responses of Cu2+ stress in roots. When treated with Zn2+, the expression of ScMT 1 and ScMT 3 increased only in shoots and roots, respectively. The expression of ScMT 2-1-5 was up-regulated at the early stage and then suppressed in both shoots and roots. It suggests that ScMT 1 and ScMT 3 mainly function in shoots and roots, respectively, when exposed to Zn2+ stress. ScMT 2-1-5 was involved in the molecular responses of Cd2+ stress at the early stage. ScMT 1, ScMT 2-1-5, and ScMT 3 showed similar or complementary expression patterns in different tissues of sugarcane when exposed to heavy metals, which revealed the functional diversity of sugarcane MTs in detoxification and cellular redox regulation, and their spatiotemporal coordination in mitigating or even preventing tissue injury caused by excessive heavy metals. The results provide a basic information for further research on mechanisms in the synergistic enhancement of tolerance to heavy metals for MTs family genes in the highly polyploid sugarcane.

Foliar Application of Zinc Oxide Nanoparticles and Zinc Sulfate Boosts the Content of Bioactive Compounds in Habanero Peppers.

The physiological responses of habanero pepper plants (Capsicum chinense Jacq.) to foliar applications of zinc sulphate and zinc nano-fertilizer were evaluated in greenhouse trials. The effect of the supplement on fruit quality of habanero pepper was particularly observed. Habanero pepper plants were grown to maturity, and during the main stages of phenological development, they were treated with foliar applications of Zn at concentrations of 1000 and 2000 mg L−1 in the form of zinc sulfate (ZnSO4) and zinc oxide nanoparticles (ZnO NPs). Additional Zn was not supplied to the control treatment plants. ZnO NPs at a concentration of 1000 mg L−1 positively affected plant height, stem diameter, and chlorophyll content, and increased fruit yield and biomass accumulation compared to control and ZnSO4 treatments. ZnO NPs at 2000 mg L−1 negatively affected plant growth but significantly increased fruit quality, capsaicin content by 19.3%, dihydrocapsaicin by 10.9%, and Scoville Heat Units by 16.4%. In addition, at 2000 ZnO NPs mg L−1 also increased content of total phenols and total flavonoids (soluble + bound) in fruits (14.50% and 26.9%, respectively), which resulted in higher antioxidant capacity in ABTS (2,2′azino-bis(3-ethylbenzothiazoline-6-sulfonic acid)), DPPH (2,2-diphenyl-1-picrylhydrazyl), and FRAP (ferric reducing antioxidant power) (15.4%, 31.8%, and 20.5%, respectively). These results indicate that application of ZnO NPs could be employed in habanero pepper production to improve yield, quality, and nutraceutical properties of fruits.

Influence of Zinc sulphate and Zinc EDTA on Grain zinc, Growth and Yield Parameters, of Rice Genotypes

A field experiment was conducted at the Department of Rice, Tamil Nadu Agricultural University, Coimbatore-3 during Kharif season, 2017. The objective of the experiment was to study the influence of Zinc nutrition on growth, physiology, ZUE, yield and grain Zn content of rice genotypes at two stages of crop. The experiment was laid out in a factorial randomized block design with three replications. The results of the field study revealed the maximum total tillers per plant were observed in the rice genotype IR93354:19-B-12-21-9-1RGA-2RGA-1-B under 0.5% ZnSO4 condition at the grain filling stage. However the higher total dry matter production recorded in the genotype IR15M1266 under 0.5% ZnSO4 treatment. Regarding zinc use efficiency indices, the maximum zinc use efficiency (42.2%) was observed in the foliar application of 0.5% ZnSO4 in rice genotype IR14M117 followed by BPT5204 (39.0%). The higher spikelet fertility was observed in BPT5204 (98.2%) under foliar application of 0.5% ZnSO4 was followed by Kalanamak (97.3%) on par with Chittmutyalu (97.2%) was under 0.5% ZnSO4 application. The treatment 0.5% ZnSO4 had recorded higher grain yield (8870.0 kg/ha) in IR95097:3-B-16-11-4-GBS which was on par with IR14M124 (8800 kg/ ha). To conclude, when foliar sprayed with zinc sources such as 0.5% ZnSO4 and 0.5 % Zn EDTA at boot leaf and grain filling stage in rice genotypes like IR15M1003, IR95097:3-B-16-11-4-GBS, IR95040:12-B-3-10-GBS, IR15M1341 and IR95097:4-B-2-18-8-GBS showed significant changes in growth, physiology and zinc grain content..

Toxicogenomic responses of Caenorhabditis elegans to pristine and transformed zinc oxide nanoparticles

Manufactured nanoparticles (MNPs) undergo transformation immediately after they enter wastewater treatment streams and during their partitioning to sewage sludge, which is applied to agricultural soils in form of biosolids. We examined toxicogenomic responses of the model nematode Caenorhabditis elegans to pristine and transformed ZnO-MNPs (phosphatized pZnO- and sulfidized sZnO-MNPs). To account for the toxicity due to dissolved Zn, a ZnSO4 treatment was included. Transformation of ZnO-MNPs reduced their toxicity by nearly ten-fold, while there was almost no difference in the toxicity of pristine ZnO-MNPs and ZnSO4. This combined with the fact that far more dissolved Zn was released from ZnO- compared to pZnO- or sZnO-MNPs, suggests that dissolution of pristine ZnO-MNPs is one of the main drivers of their toxicity. Transcriptomic responses at the EC30 for reproduction resulted in a total of 1161 differentially expressed genes. Fifty percent of the genes differentially expressed in the ZnSO4 treatment, including the three metal responsive genes (mtl-1, mtl-2 and numr-1), were shared among all treatments, suggesting that responses to all forms of Zn could be partially attributed to dissolved Zn. However, the toxicity and transcriptomic responses in all MNP treatments cannot be fully explained by dissolved Zn. Two of the biological pathways identified, one essential for protein biosynthesis (Aminoacyl-tRNA biosynthesis) and another associated with detoxification (ABC transporters), were shared among pristine and one or both transformed ZnO-MNPs, but not ZnSO4. When comparing pristine and transformed ZnO-MNPs, 66% and 40% of genes were shared between ZnO-MNPs and sZnO-MNPs or pZnO-MNPs, respectively. This suggests greater similarity in transcriptomic responses between ZnO-MNPs and sZnO-MNPs, while toxicity mechanisms are more distinct for pZnO-MNPs, where 13 unique biological pathways were identified. Based on these pathways, the toxicity of pZnO-MNPs is likely to be associated with their adverse effect on digestion and metabolism.

Selective Inhibition of HIF1α Expression by ZnSO4 Has Antitumoral Effects in Human Melanoma.

Zinc as an essential trace metal is a ubiquitous component of various molecules of the cell. Studies indicated that it may modulate functions of various cancer cell types, and can even inhibit metastasis formation in experimental models. In melanoma, zinc was shown to affect melanin production and to induce apoptosis. Using human melanoma cell lines, we have tested the effects of ZnSO4 on cell proliferation, survival, migration as well as in vivo on experimental liver colony formation. We have found that ZnSO4 has antiproliferative and proapoptotic effects in vitro. In SCID mice intraperitoneal administration of ZnSO4 specifically inhibited liver colony formation without affecting primary tumor growth. To reveal the molecular mechanisms of action of zinc in human melanoma, we have tested mRNA expression of zinc finger transcription factors and found a strong inhibitory effect on HIF1α, as compared to WT1 whereas HIF2α and MTF1 expression was unaffected. Immunohistochemical detection of HIF1α protein in liver metastases confirmed its decreased nuclear expression after in vivo ZnSO4 treatment. These data indicate that in human melanoma zinc administration may have an antimetastatic effect due to a selective downregulation of HIF1α.

Effect of ZnSO4 fertilizer on soil chemical properties, performance and proximate quality of sweet potato in a derived savanna ecology of Nigeria

Abstract Micro-nutrients especially zinc can not only increase the yield of sweet potato but can also improve the quality of tubers. Hence, experiments were carried out in 2015 and 2016 cropping seasons to determine the impact of various levels of ZnSO4 fertilizer on soil chemical properties, foliage and storage root yields and proximate qualities of sweet potato (Ipomoea batatas L.). The experiments consisted of 5 levels (0, 5, 10, 15 and 20 kg ha-1) of ZnSO4 fertilizer. These were arranged in a randomized complete block design and replicated three times. ZnSO4 increased (with the exception of P) soil chemical properties compared with the control. N, K, Ca, Mg and Zn were increased up to the 20 kg ha-1 ZnSO4 level in both years. ZnSO4 reduced P concentrations in soil as the level increased. For sweet potato performance, 5 kg ha-1 ZnSO4 fertilizer had the highest values of foliage yield (vine length and vine weight) and storage root yield. Using the mean of the two years and compared with the control, ZnSO4 fertilizer at 5 kg ha-1 increased storage root yield of sweet potato by 17.4%. On fitting the mean storage root yield data of the two years with a cubic equation, the optimum rate of Zn for sweet potato was found to be 3.9 kg ha-1 to achieve the maximum sweet potato yield. In this study, relative to the control, ZnSO4 fertilizer increased moisture and decreased the fibre contents of sweet potato. There were no consistent patterns of variation between the 5, 10, 15 and 20 kg ha-1 ZnSO4 treatments for proximate qualities except that the highest values of fat, protein, carbohydrate and ash was at 5 kg ha-1 ZnSO4.

Heterologous expression of a Glyoxalase I gene from sugarcane confers tolerance to several environmental stresses in bacteria.

Glyoxalase I belongs to the glyoxalase system that detoxifies methylglyoxal (MG), a cytotoxic by-product produced mainly from triose phosphates. The concentration of MG increases rapidly under stress conditions. In this study, a novel glyoxalase I gene, designated as SoGloI was identified from sugarcane. SoGloI had a size of 1,091 bp with one open reading frame (ORF) of 885 bp encoding a protein of 294 amino acids. SoGloI was predicted as a Ni2+-dependent GLOI protein with two typical glyoxalase domains at positions 28–149 and 159–283, respectively. SoGloI was cloned into an expression plasmid vector, and the Trx-His-S-tag SoGloI protein produced in Escherichia coli was about 51 kDa. The recombinant E. coli cells expressing SoGloI compared to the control grew faster and tolerated higher concentrations of NaCl, CuCl2, CdCl2, or ZnSO4. SoGloI ubiquitously expressed in various sugarcane tissues. The expression was up-regulated under the treatments of NaCl, CuCl2, CdCl2, ZnSO4 and abscisic acid (ABA), or under simulated biotic stress conditions upon exposure to salicylic acid (SA) and methyl jasmonate (MeJA). SoGloI activity steadily increased when sugarcane was subjected to NaCl, CuCl2, CdCl2, or ZnSO4 treatments. Sub-cellular observations indicated that the SoGloI protein was located in both cytosol and nucleus. These results suggest that the SoGloI gene may play an important role in sugarcane’s response to various biotic and abiotic stresses.

Influence of late season foliar application of urea, boric acid and zinc sulfate on nitrogenous compounds concentration in the bud and flower of Hayward kiwifruit

Abstract Proper management of late season mineral nutrition and improving nitrogenous reserves in kiwifruit vines can result in efficient growth and yield in the following season. This experiment investigated the influence of late season foliar application type and time on the content of total nitrogen, total protein, total free amino acid, and individual amino acids of bud and flower and consequently vegetative and reproductive growth of Hayward kiwifruit in next spring. The orchard was located in Vajargah, Guilan province, Iran, with an average annual rainfall and temperature 1147 mm and 15.7 °C, respectively. Foliar application of mineral nutrients included urea (0.25%, 0.5% and 1%), zinc sulfate (1000, 1500 and 2000 mg l−1) and boric acid (500, 1000 and 1500 mg l−1) alone and in combined treatments as urea (0.25%) + H3BO3 (500 mg l−1) + ZnSO4 (1000 mg l−1); urea (0. 5%) + H3BO3 (1000 mg l−1) + ZnSO4 (1500 mg l−1); urea (1%) + H3BO3 (1500 mg l−1) + ZnSO4 (2000 mg l−1). The experimental treatments were conducted in 8 years old kiwifruit vines in late growing season. To determine appropriate time for late season foliar application, fertilization was performed in middle September, early and late October, 96, 75 and 54 days before the beginning of the winter dormancy, respectively. Samples were taken from buds at two stages: before leaf abscission and again during dormancy. Flower sampling was taken at the full bloom stage. Results indicated that 33.65 and 35.75 percentages of increases in dormant bud and flower nitrogen concentrations were achieved by combined foliar application of urea (1%), zinc sulfate (2000 mg l-1) and boric acid (1500 mg l-1) compared to control vines. Moreover, urea (1%) + H3BO3 (1500 mg l−1) + ZnSO4 (2000 mg l−1) treatment increased total protein, free amino acid and most individual amino acid concentration in analyzed tissue, however this significantly consistent elevation in almost all measured parameters was not found in other fertilization treatments. Interestingly, this treatment improved 14.51% leaf area and 22.21% fruit seed number in next growing season compared to control vines. Considering all measured parameters, late October was the best late season foliar application time. Based on our findings, late season mineral foliar application, especially urea (1%) + H3BO3 (1500 mg l−1) + ZnSO4 (2000 mg l−1), in late October can improve all nitrogenous compounds concentration in bud and flower tissues, as well as increase seed number and consequently fruit weight and grower income in the next growing season.

Foliar nutrient applications to ‘Wonderful’ pomegranate (Punica granatum L.). II. Effects on leaf nutrient status and fruit split, yield and size

Abstract Though pomegranates are beginning to be farmed more intensively, little is known about pomegranate leaf nutrient concentrations or the effects of foliar nutrient applications on those concentrations, including for one of the industry’s standard cultivars, Wonderful. Foliar nutrient applications could potentially be used in commercial pomegranate production, not only to improve plant nutrient status, but also to prevent physiological disorders, such as pomegranate fruit split, the most important physiological disorder in pomegranate production. Pomegranate trees at two ‘Wonderful’ commercial orchards were treated with foliar applications of zinc sulfate (ZnSO4; 0.3%, 0.4%, and 0.5%), magnesium sulfate (MgSO4; 1%, 2%, and 3%), potassium nitrate (KNO3; 1%, 2%, and 3%), or deionized (DI) water (control). Leaf mineral nutrient concentrations were determined and fruit were analyzed for fruit split incidence, yield, fruit number per tree, fruit diameter, fruit mass, aril mass, and mass of 100 arils. Leaf Zn concentrations were significantly higher in response to all foliar ZnSO4 treatments. Foliar applications of KNO3 resulted in significant increases in leaf N and K concentrations and foliar MgSO4 resulted in a significant increase in leaf S concentration. All foliar fertilizer types resulted in significantly lower fruit split incidence at one of the study sites. The foliar fertilizer treatments did not have a significant effect on fruit number per tree, fruit diameter and mass, mass of all arils in fruit, or mass of 100 arils. The results suggest that foliar ZnSO4, MgSO4, and KNO3 could be used to improve pomegranate nutrient availability and to decrease pomegranate fruit split incidence without negatively impacting fruit yield or size of ‘Wonderful’ pomegranate.

Effect of Gibberellic acid and Zinc sulphate on Vegetative, Flowering, Seed yield and Chemical consistent of jojoba plant (Simmondsia chinensis)

This experiment was conducted to study jojoba response (Simmondsia chinensis) evergreen dioecious shrub to zinc sulphate plus gibberellic acid foliar application on vegetative, flowering, seed yield and seed chemical composition at private farm during two successive seasons of 2015 and 2016 in the Egypt. An experiment was laid out in randomized complete block design with three replications. In this study, jojoba plants were sprayed with all combination treatments of Zinc sulphate (0, 25, 50 and 75ppm) plus gibberellic acid (0, 50, 100 and 150ppm) thrice in the beginning of December, March and May. There results revealed that all combination treatments showed a significant improvement in all examined parameters with an increase in ZnSO4 / GA3 levels in comparison with untreated trees. Therefore, the maximum significant branch length (99.36 and 103.46 cm), secondary branches length (55.82 and 58.36 cm) obtained by application of 75ppm ZnSO4 plus 150ppm GA3 treatment, so this combination recorded the highest percentage of flowering %, final fruit set (95.01, 95.24%), total chlorophyll, mineral% content, seed yield per feedan (2200, 2145 kg) and seed lipid content(57.6%, 58.55%) at first and second terms respectively. The application of 75ppm ZnSO4 plus 150ppm GA3 treatment is recommended to improve jojoba traits which lead to raise its economic value as a promising tree which potentially useful as a biofuel with multi chemical and pharmaceuticals industries uses.

Effect of Foliar Zinc Application on Bioaccessibility of Cadmium and Zinc in Pakchoi

Two cultivars of pakchoi with different cadmium (Cd) accumulation were grown in nutrient solutions containing low and sufficient zinc (Zn) levels. ZnSO4 and ZnNa2 EDTA were applied as foliar fertilizers. The bioaccessibilities of Cd and Zn in pakchoi were assessed by the in vitro digestion method, and the bioaccessible established daily intakes (BEDI) of Cd and Zn from pakchoi were calculated. The effect of foliar zinc application on concentrations, bioaccessibilities, and BEDIs of Cd and Zn in pakchoi was evaluated. Results show that the Cd concentrations in shoots of the tested pakchoi cultivars under sufficient Zn condition were significantly lower than those under low Zn condition, and foliar application of ZnNa2 EDTA significantly decreased the Cd concentrations of pakchoi. The tested pakchoi cultivars with sufficient Zn had a significantly higher mean shoot Zn concentration than those with low Zn. Foliar Zn treatments significantly increased shoot Zn concentrations of pakchoi, with the highest in the ZnSO4 treatment. Cd bioaccessibility in the tested pakchoi cultivars with sufficient Zn was significantly lower than that with low Zn. Foliar applied Zn could significantly reduce Cd bioaccessibility in the gastric phase, with a maximal reduction of 35.81% compared to the control. Foliar treatment with ZnSO4 could significantly decrease Cd bioaccessibility in the small intestinal phase, with a maximal reduction of 59.24% compared to the control. Foliar Zn treatments reduced significantly the Zn bioaccessibility of pakchoi in the gastric and small intestinal phases, with a maximal reduction of 68.90% compared to the control. The reduction of Zn bioaccessibility was higher in the ZnSO4 treatment than in the ZnNa2 EDTA treatment. Via the consumption of the Cd-contaminated common cultivar Hanlv, the BEDI values of Cd were higher than that of the provisional tolerable daily intake (PTDI) standard of WHO [0.83 μg·(kg·d)-1]. However, via the consumption of the low-Cd accumulating cultivar Huajun or pakchoi with foliar application of ZnSO4, the BEDI values of Cd decreased significantly and were below the PTDI value. Via the consumption of pakchoi in the control or low-level ZnNa2 EDTA treatments, the BEDI values of Zn were below that of the recommended nutrient intake for Zn. However, via the consumption of pakchoi with foliar application of ZnSO4 or high-level ZnNa2 EDTA, the BEDI values of Zn were higher than that of the recommended nutrient intake and met the human needs for Zn from vegetables. Under the ZnSO4 treatment, the BEDI values of Zn from pakchoi were the highest. In conclusion, foliar zinc application could significantly reduce the bioaccessibilities of Cd and Zn in pakchoi and the BEDI values of Cd and increase the BEDI values of Zn. Foliar application with ZnSO4 was the most suitable treatment to reduce Cd intake and increase Zn intake from pakchoi.

ZnSO4 rescued vimentin from collapse in DBP-exposed Sertoli cells by attenuating ER stress and apoptosis

Sertoli cells (SCs) provide physical and nutritional support for spermatogenesis. Dibutyl phthalate (DBP) is a plasticizer that has male reproductive toxicity. The collapse of vimentin in DBP-exposed SCs is thought to induce the sloughing of spermatocytes from seminiferous tubules. In this study, we explored methods to rescue vimentin from collapse in DBP-exposed SCs. DBP not only induced the hyperphosphorylation of vimentin but also triggered endoplasmic reticulum (ER) stress and apoptosis in SCs. Treatment with BAPTA-AM, an antagonist of Ca2+, significantly decreased the level of phosphorylated vimentin, while LY294002, an inhibitor of Akt1, did not. ER stress and apoptosis remained at high levels, and the distribution of vimentin was not improved. ZnSO4 treatment did not decrease the level of phosphorylated vimentin. However, after treatment, ER stress and apoptosis were obviously inhibited, and the distribution of vimentin was reconverted. These results indicated that ZnSO4 could alleviate the collapse of vimentin by attenuating ER stress and apoptosis. This study suggested that an appropriate zinc supply might be a choice to alleviate DBP-induced adverse reproductive effects.

Alleviation of nickel toxicity and an improvement in zinc bioavailability in sunflower seed with chitosan and biochar application in pH adjusted nickel contaminated soil

ABSTRACTThis study demonstrates the reduction of nickel (Ni) toxicity and enhancing zinc (Zn) bioavailability from sunflower seed with biochar (BC) and chitosan (CH) application to Ni contaminated soil. The pH of experiment soil was adjusted to value 6.5 prior to its use in a greenhouse experiment. Acidified Ni contaminated soil was amended with Zn (4 mg kg−1 soil), while BC and CH, both as alone [1% each (w/w)] and in combination [0.5% each (w/w)], were used as immobilizing agents for Ni. Significantly higher concentrations of Zn were found in seed receiving ZnSO4 treatments. However, these results were more pronounced in the treatment (Zn+CH+BC) where Zn, CH, and BC were together applied. The Zn+CH+BC treatment improved seed quality parameters, phytate/minerals molar ratios, increased seed weight (per pot) up to 75%, Fe 41%, Zn 42% in seed, while reduced Ni concentration up to 17% in seed, over control. Similarly, Zn+CH+BC treatment significantly decreased oxidative stress by improving antioxidant defense system in sunflower. The results of this preliminary study suggest that further studies are obligatory to corroborate the extent of Ni immobilization in soil and enhancing Zn bioavailability from sunflower seed under field conditions with realistic and applicable rates of BC and CH.

Zinc Sulphate Mediates the Stimulation of Cell Proliferation of Rat Adipose Tissue-Derived Mesenchymal Stem Cells Under High Intensity of EMF Exposure.

Unlike the role of mesenchymal stem cells (MSCs) in regenerative medicine, their application in cell therapy can be complicated by factors such as a reduction in the proliferation potential, senescent tendency of MSCs to expand, and an age-dependent decline in their number and functions. It was shown that there is an association between exposure to electromagnetic fields (EMFs) and response to stress, cell proliferation, aging, and cell death. Furthermore, the zinc ion, as an essential trace element, was reported to be involved in the regulation of the growth and cell proliferation. In this report, ratadipose tissue-derived mesenchymal stem cells (rADSCs) were randomly divided into four groups-group I (control without any ZnSO4 and EMF); group II (ZnSO4 treatment without EMF exposed); group III (EMF exposed without ZnSO4); and group ІV (EMF exposed with ZnSO4)-to evaluate whether 0.14μg/ml Zinc sulfate (ZnSO4) could affect cell proliferation of rADSCs under extremely low frequency-electromagnetic field (ELF-EMF). The methyl thiazoltetrazolium (MTT) method was used to determine the cell proliferation of rADSCs following exposure to ELF-EMF in the presence of ZnSO4. The immunocytochemistry method as well as flow-cytometry was used to identify the cell surface markers. Next, oil red O, alizarin red, toluidine blue, and cresyl violet staining was done to evaluate the adipogenic, osteogenic, chondrogenic, and neurogenic differentiation of rADSCs as the pluripotent capacity of rADSCs, respectively. The results showed that an exposure to ELF-EMF caused a decrease in the proliferation of rADSCs. However, the ZnSO4 supplementation significantly increased the cell proliferation of ELF-EMF-exposed rADSCs. In addition, in the presence of 0.14μg/ml ZnSO4, rADSCs appeared to be growing faster than the control group and ZnSO4 significantly decreased the doubling time of ELF-EMF-exposed rADSCs. It seems that ZnSO4 would be a good element to induce the cell proliferation of ELF-EMF-exposed rADSCs.

Seed priming improves seedling emergence and reduces oxidative stress in Nigella sativa under soil moisture stress

ABSTRACTNigella sativa seeds have multiple industrial and pharmaceutical uses for the diseases treatment such as bronchitis, rheumatism, high blood pressure, cough, eczema, inflammation, and influenza. Although semi-arid areas are suitable for production of annual medicinal plants because they receive favorable amounts of solar radiations, the establishment of these plants' seedling requires frequent irrigations because their seeds are small and the climate is hot at the time of planting. When the irrigation sources are limited, poor establishment and inadequate production will result. Therefore, in order to improve nitrogen (N) in N. sativa seedling establishment, different effects of seed priming treatments were evaluated under drought stress. Different levels of soil moisture (irrigation after 11, 22, 33, and 44 mm of evaporation as no water stress, mild water stress, moderate water stress, and severe water stress, respectively) and seed priming treatments [no priming, potassium nitrate (KNO3), zinc sulfate (ZnSO4), polyethylene glycol 6000, and gibberellic acid (GA)] were evaluated as the main plot and subplot factors, respectively. The results showed that under mild water stress condition, seed priming significantly increased proline content and the highest percentage of emergence was obtained with ZnSO4. Furthermore, the highest amount of soluble protein was observed in the KNO3 treatment and the catalase enzyme content increased in all priming treatments, except polyethylene glycol treatment. Under moderate water stress condition, the KNO3 treatment raised the rate and percentage of emergence and the seedling length. However, under severe water stress condition, the highest percentage of emergence was achieved by GA and ZnSO4 treatments. Moreover, hydropriming and GA produced the highest proline content and seedling dry weight. Under moderate and severe water stress conditions, the hydropriming and ZnSO4 treatments showed a significant increase in catalase activity. It can be concluded that priming of N. sativa seeds with ZnSO4 and GA can alleviate drought stress in the early stages of seedling growth by increasing the antioxidants in the seedlings. Farmers in semi-arid regions can prime N. sativa seeds with these compounds before planting to increase production of this valuable medicinal plant and reduce water consumption.

Zinc Sulfate Foliar Application Effects on Some Physiological Characteristics and Phenolic and Essential Oil Contents of Lavandula stoechas L. Under Sodium Chloride (NaCl) Salinity Conditions

ABSTRACTLavandula stoechas L. plant is a perennial evergreen used as a fragrant ornamental and medicinal plant. In order to study the effect of foliar spray of zinc sulfate (ZnSO4) (0, 1000, and 2000 mgL−1) and sodium chloride (NaCl) salinity (0, 75, and 150 mM) on some physiological characteristics of Lavandula stoechas L. plants, a pot experiment was carried out at the Research Greenhouse of Azarbaijan Shahid Madani University, Iran. Physiological characteristics [root and leaf dry weights, total soluble solids (TSSs), total anthocyanins, chlorophyll b, Zn2+, potassium (K+) contents, and K+/Na+ ratio] were significantly affected by the interaction effects of ZnSO4 foliar application and salinity levels. The highest root dry weight, chlorophyll b, anthocyanin, and Zn contents as well as TSS were found in the plants with NaCl0 × ZnSO4 2000 mgL−1. For Na+, the greatest value was recorded with NaCl 150 × ZnSO40. The highest K+/Na+ ratio was found in the control plants. Foliar application of ZnSO4 promoted the total phenolic content, especially at 2000 mgL–1. The highest amounts of flower dry weights were recorded at NaCl0. Chlorophyll a, total flavonoids, stem dry weight, and essential oil content were affected with ZnSO4 treatment and salinity levels. The highest values for the essential oil content, chlorophyll a content, and stem dry weight were attained by the nonsaline treatment. Both foliar application levels positively influenced the essential oil and flavonoid contents of the plants. The results reveal that zinc application had marked effects on the physiological characteristics of Lavandula stoechas L. plants growing under salinity stress conditions.

Zinc oxide nano particles increases Zn uptake, translocation in rice with positive effect on growth, yield and moisture stress tolerance

Zinc (Zn) is one of the important micronutrients required for plant growth and development. Soil Zn deficiency is a major problem and hence adequate Zn supply during crop developmental stages are recommended to improve the nutrient content in the edible parts and also improve productivity. However, excess use of traditional Zn fertilizers has detrimental effect on soil and plant health. Hence, there is a need to have formulations of Zn with improved use efficiency for the better crop performance with less input. In this context, we investigated the impact of Zn Oxide (ZnO) nano particles on Zn uptake, translocation and growth performance in rice in comparison to conventional Zn fertilizer (ZnSO4). Seed priming with 1000 ppm of ZnO was found to be optimal for increasing seedling vigour index. Significant improvement in growth of pot grown rice plants was observed with ZnO treatment compared to ZnSO4, particularly with seed priming followed by foliar application. Plants treated with nano ZnO showed improved plant height, chlorophyll content, biomass, tiller number and yield. The Zn content in leaf and seed is also higher in ZnO nano treated plant samples compared to ZnSO4 treatment. With increased Zn levels in leaves, substantial up regulation of Cu/Zn SOD was observed. Under moisture stress conditions, the ZnO nano particles application improved the tolerance by maintaining the membrane stability and showed higher expression of Cu/Zn SOD. This signifies that optimum levels of Zn enhances cell metabolism leading to improved growth and stress tolerance. Overall the study demonstrates the positive effects of ZnO nano particle in improving the plant growth.

ZINC PROMOTES FOCAL CEREBRAL ISCHEMIA-REPERFUSION INJURY THROUGH ACTIVATING CDK5 BY TYR15 PHOSPHORYLATION

Aberrant CDK5 activation plays a key role in mediating ischemic stroke injury, while the underlying mechanism for CDK5 activation remains incompletely clarified. Hippocampal CA1 neurons are highly sensitive to ischemic injury, it was discovered that a large number of ionic zinc accumulated in that area after ischemia-reperfusion. Whether zinc is involved in CDK5 activation in ischemia-reperfusion injury and the regulation pathway has not been elucidated. MCAO rat model was established, with or without intraperitoneal injection of zinc chelator CQ or intraventricular injection of tyrosine kinase SFK inhibitor PP2 before embolization. After 6 hours of reperfusion, rat were sacrificed and brains were separated. TTC staining was performed to detect the area of brain infarction. The protein levels of total CDK5, phosphorylated CDK5 at Y15, p25 and p35 were detected by immunoblotting. The activity of CDK5 was detected by using Histone H1 as substrate. At the cellular level, N2a cells were incubated with ZnSO4 for 3 hours, with or without pretreatment with CQ or PP2. The protein test wan same as before. GFP-tagged wild-type or Y15F-mutated CDK5 were overexpressed in N2a cells and then cells were incubated with ZnSO4. After the treatment, exogenously expressed CDK5 was pull-down by using anti-GFP antibody and CDK5 activities were measured. In the rat MCAO model, CDK5 was activated by ischemia-reperfusion injury, and the level of Y15 phosphorylated CDK5 was increased remarkably. CQ or PP2 pretreatment significantly decreased the brain infarction area, reversed neuron loss, CDK5 activation and CDK5-Y15 phosphorylation, but not p35 cleavage to p25. At the cellular level, ZnSO4 treatment resulted in CDK5 activation, and CDK5-Y15 phosphorylation, which could also be reversed by CQ or PP2. The unphosphorylated form of CDK5 – Y15F mutant could not be phosphorylated and activated by zinc treatment. Activation of CDK5 by zinc was not dependent on p35 cleaved into p25 pathway. During stroke, cellular released zinc activates the tyrosine protein kinase, which further leads to Y15 phosphorylation and activation of CDK5, thus mediates neuronal damage. The CDK5 activation does not depend on p35 cleavage. These results revealed a novel mechanism of abnormal activation of CDK5 during ischemia-reperfusion injury.

Effects of Maternal Zinc Glycine on Mortality, Zinc Concentration, and Antioxidant Status in a Developing Embryo and 1-Day-Old Chick.

This study was conducted to investigate the effects of maternal zinc glycine (Zn-Gly) supplementation as an alternative for zinc sulfate (ZnSO4) on mortality, zinc (Zn) concentration, and antioxidant status in a developing embryo and 1-day-old chick. Six hundred 39-week-old broiler breeders were randomly assigned to 6 treatments, each treatment including 5 replicates with 20 birds each. Six treatments received a basal diet (control, 24mg Zn/kg diet) or a basal diet supplemented with ZnSO4 (80mg Zn/kg) or Zn-Gly (20, 40, 60, or 80mg Zn/kg), respectively. The experiment lasted for 8weeks after a 4-week pre-experiment with a basal diet. At the last week, 100 eggs per replicate were randomly collected for incubation. Compared with the control treatment, Zn supplementation decreased (P<0.05) embryo mortalities of the late stage and the whole period, increased (P<0.05) liver Zn concentration in the embryo of d9, d19, and 1-day-old chick, and improved (P<0.05) antioxidant status in the embryo of d19 and 1-day-old chick. Compared with the ZnSO4 treatment, 80mg Zn/kg Zn-Gly treatment significantly decreased (P<0.05) the late stage embryo mortality and increased (P<0.05) liver Zn concentration in the embryo of d9, d19, and 1-day-old chick. The 80mg Zn/kg Zn-Gly treatment significantly increased (P<0.05) copper-zinc superoxide dismutase activity in d19 embryo and 1-day-old chick, total superoxide dismutase activity in 1-day-old chick, and copper-zinc superoxide dismutase messenger RNA (mRNA) abundance of d9 embryo and 1-day-old chick than that in ZnSO4 treatment. The liver metallothionein concentration of the developing embryo and 1-day-old chick and its mRNA abundance of d19 embryo were also significantly increased (P<0.05) in the 80mg Zn/kg Zn-Gly treatment in comparison with ZnSO4 treatment. In conclusion, maternal Zn supplementation decreased embryo mortalities of the late stage and the whole period by increasing liver Zn concentration and antioxidant status in d19 embryo and 1-day-old chick, and 80mg Zn/kg from Zn-Gly treatment was the optimum choice.

Newly developed method for mouse olfactory behavior tests using an automatic video tracking system

ObjectiveThe mouse is the most popular animal model in olfactory research. Behavior tests with odorants are essential for determining olfactory phenotype. To the best of our knowledge, the mouse olfactory behavior test has not been standardized, making the results vulnerable to inter-observer variation. We sought to develop a new mouse olfactory behavior test assessed by an automatic video tracking system with minimal inter-observer variation. MethodsA video-tracking system was used to automatically track mouse behavior in standard breeding cages with C57BL/6N mice. We tested two odorants (peanut butter for the preference test, 2MB acid for the avoidance test) and distilled water (for a control). Mouse behavior was recorded for 3min and analyzed. For the preference test, investigation time was measured. For the avoidance test, time spent in sectors away from the odorant zone was measured. To confirm our experimental settings, we also evaluated an anosmia mouse model prepared with intranasal administration of ZnSO4. ResultsAll strains of mice showed reproducible behavior patterns of preference or avoidance for the odorants. The anosmia mouse model, as expected, failed to show an olfactory ability for preference or avoidance, and this was well-matched by histologic changes caused by the ZnSO4 treatment. The automatic video tracking system successfully tracked and automatically calculated mouse behavior with good reproducibility. ConclusionOur olfactory behavior test offers a simple and accurate method to evaluate olfactory function in mice. This test can be utilized as a possible standard method to search for features of olfactory phenotypes in mice.