Populus euphratica R2R3-MYB transcription factor RAX2 binds ANN1 promoter to increase cadmium enrichment in Arabidopsis.

The expression of R2R3-MYB transcription factor PeRAX2 increased transiently upon CdCl2 exposure (100 µM, 48 h) in leaves and roots of Populus euphratica. We observed that overexpression of PeRAX2 increased Cd2+ concentration in Arabidopsis root cells and Cd2+ amount in whole plant, which was due to the increased Cd2+ influx into root tips. However, the Cd2+ influx facilitated by PeRAX2 overexpression was substantially reduced by LaCl3 (an inhibitor of Ca2+-channels), suggesting that PeRAX2 could promote the Cd2+ entering through PM Ca2+-permeable channels (CaPCs) in the roots. It is noting that the expression of annexin1 (AtANN1), which mediates the influx of divalent cations through the PM calcium channels, was upregulated by Cd2+ in PeRAX2-transgenic Arabidopsis. Bioinformatic analysis revealed that the AtANN1 promoter (AtANN1-pro) contains four cis-elements for MYB binding. The PeRAX2 interaction with AtANN1-pro was validated by LUC reporter assay, EMSA, and Y1H assay. Our data showed that PeRAX2 binds to the AtANN1 promoter region to regulate gene transcription and that AtANN1 mediates the Cd2+ entry through CaPCs in the PM, leading to a Cd2+ enrichment in transgenic plants. The PeRAX2-stimulated Cd2+ enrichment consequently resulted in high H2O2 production in root cells of transgenic plants. The expression of AtSOD and AtPOD and activities of CAT, SOD, POD increased in the transgenic lines under Cd2+ stress. However, the Cd2+-upregulated expression and activity of antioxidative enzymes were less pronounced in the PeRAX2-overexpressed lines, compared to the wildtype and vector controls. As a result, root length and plant growth were more suppressed by Cd2+ in the transgenic lines. Our data suggest that transcriptional regulation of AtANN1 by PeRAX2 can be utilized to improve Cd2+ enrichment and phytoremediation, although the enriched Cd2+ affected antioxidant defense system and plant growth in the model species.

Vascular endothelial dysfunction induced by 3-bromofluoranthene via MAPK-mediated-NFκB pro-inflammatory pathway and intracellular ROS generation.

3-Bromofluoranthene (3-BrFlu) is the secondary metabolite of fluoranthene, which is classified as a polycyclic aromatic hydrocarbon, through bromination and exists in the fine particulate matter of air pollutants. Endothelial dysfunction plays a critical role in the pathogenesis of cardiovascular and vascular diseases. Little is known about the molecular mechanism of 3-BrFlu on endothelial dysfunction in vivo and in vitro assay. In the present study, 3-BrFlu included concentration-dependent changes in ectopic angiogenesis of the sub-intestinal vein and dilation of the dorsal aorta in zebrafish. Disruption of vascular endothelial integrity and up-regulation of vascular endothelial permeability were also induced by 3-BrFlu in a concentration-dependent manner through pro-inflammatory responses in vascular endothelial cells, namely, SVEC4-10 cells. Generation of pro-inflammatory mediator PGE2 was induced by 3-BrFlu through COX2 expression. Expression of COX2 and generation of pro-inflammatory cytokines, including TNFα and IL-6, were induced by 3-BrFlu through phosphorylation of NF-κB p65, which was mediated by phosphorylation of MAPK, including p38 MAPK, ERK and JNK. Furthermore, generation of intracellular ROS was induced by 3-BrFlu, which is associated with the down-regulated activities of the antioxidant enzyme (AOE), including SOD and catalase. We also found that 3-BrFlu up-regulated expression of the AOE and HO-1 induced by 3-BrFlu through Nrf-2 expression. However, the 3-BrFlu-induced upregulation of AOE and HO-1 expression could not be revised the responses of vascular endothelial dysfunction. In conclusion, 3-BrFlu is a hazardous substance that results in vascular endothelial dysfunction through the MAPK-mediated-NFκB pro-inflammatory pathway and intracellular ROS generation.

Selenium increases antimony uptake in ramie (Boehmeria nivea L.) by enhancing the physiological, antioxidative, and ionomic mechanisms.

Ramie (Boehmeria nivea L.) is a promising phytoremediation candidate due to its high tolerance and enrichment capacity for antimony (Sb). However, challenges arise as Sb accumulated mainly in roots, complicating soil extraction. Under severe Sb contamination, the growth of ramie may be inhibited. Strategies are needed to enhance Sb accumulation in ramie's aboveground parts and improve tolerance to Sb stress. Considering the beneficial effects of selenium (Se) on plant growth and enhancing resistance to abiotic stresses, this study aimed to investigate the potential use of Se in enhancing Sb uptake by ramie. We investigated the effects of Se (0.5, 1, 2, 5, or 10 µM) on ramie growth, Sb uptake and speciation, antioxidant responses, and ionomic profiling in ramie under 10 mg/L of SbIII or antimonate (SbV) stresses. Results revealed that the addition of 0.5 µM Se significantly increased shoot biomass by 75.73% under SbIII stress but showed minimal effects on shoot and root length in both SbIII and SbV treatments. Under SbIII stress, 2 µM Se significantly enhanced Sb concentrations by 48.42% in roots and 62.88% in leaves. In the case of SbV exposure, 10 µM Se increased Sb content in roots by 42.57%, and 1 µM Se led to a 91.74% increase in leaves. The speciation analysis suggested that Se promoted the oxidation of SbIII to less toxic SbV to mitigate Sb toxicity. Additionally, Se addition effectively minimized the excess reactive oxygen species produced by Sb exposure, with the lowest malondialdehyde (MDA) content at 0.5 µM Se under SbIII and 2 µM Se under SbV, by activating antioxidant enzymes including superoxide dismutase, catalase, peroxidase, and glutathione peroxidase. Ionomic analysis revealed that Se helped in maintaining the homeostasis of certain nutrient elements, including magnesium, potassium (K), calcium (Ca), iron (Fe), and copper (Cu) in the SbIII-treated roots and K and manganese (Mg) in the SbV-treated roots. The results suggest that low concentrations of Se can be employed to enhance the phytoremediation of Sb-contaminated soils using ramie.

Toxicity and physiological interruptions of a proteinaceous toxin from Metarhizium anisopliae against the olive fruit pest, Bacterocera oleae (Diptera: Tephritidae).

The use of natural bioactive compounds mainly proteinaceous secondary metabolites of fungi is one of the promising pest control methods because of their lethal effects on insects in low concentration, limited persistence in environment and easily decomposition into environmentally safe compounds. The olive fruit fly, Bactrocera oleae (Rossi) (Diptera: Tephritidae), is a destructive pest of olive fruits around the world. In the current study, the proteinaceous compounds were extracted from the two isolates (MASA and MAAI) of Metarhizium anisopliae and their effects were evaluated on toxicity, feeding performance and antioxidant system of the adult's olive flies. Both extracts from MASA and MAAI showed entomotoxicity against the adults by 2.47 and 2.38 mg/mL as LC50 concentrations. Also, LT50 values were recorded 1.15 and 1.31 days for MASA and MAAI, respectively. No statistical differences were recorded in the consumption rate of the adults on control and secondary metabolite contained protein hydrolysate. In contrast, the adults fed on LC30 and LC50 concentrations of MASA and MAAI demonstrated significant reduction in the activities of digestive alpha-amylase, glucosidases, lipase, trypsin, chymotrypsin, elastase, amino- and carboxypeptidases. Activity of antioxidant enzymes changed in the adults of B. oleae fed on the fungal secondary metabolites. Catalase, Peroxidase and Superoxide dismutase elevated in the treated adults with the highest amounts of MAAI. Similar results were found in activity of ascorbate peroxidase and glucose-6-phosphate dehydrogenase except for malondialdehyde amount in which no statistical differences were recorded between treatments and control. Relative gene expression of caspase enzymes revealed the higher expression in the treated B. oleae compared to control with the highest level of caspase 8 for MASA and caspases 1 and 8 for MAAI. Results of our study showed that the secondary metabolites extracted from the two isolates of M. anisopliae caused mortality, interrupted digestion and induced oxidative stress in the adults of B. oleae.

Impact of Beauveria bassiana on antioxidant enzyme activities and metabolomic profiles of Spodoptera frugiperda.

Spodoptera frugiperda is a pest that poses a serious threat to the production of food and crops. Entomopathogenic fungi, such as Beauveria bassiana, have shown potential for S. frugiperda control. However, the mechanism of this biological control of pathogens is not fully understood, such as how antioxidant enzyme activities and metabolic profiles in S. frugiperda larvae are affected when infected by entomopathogenic fungi. This study assessed the antioxidant enzyme activities and shift in metabolomic profile in the S. frugiperda larvae infected with B.bassiana. The results indicate a pattern of initial increase and subsequent decrease in the activities of superoxide dismutase, catalase, and peroxidase in the B.bassiana-infected larvae. And the enzyme activities at 60 h of infection ended significantly lower than those of the uninfected larvae. A total of 93 differential metabolites were identified in the B.bassiana-infected larvae, of which 41 metabolites were up-regulated and 52 were down-regulated. These metabolites mainly included amino acids, nucleotides, lipids, carbohydrates, and their derivatives. Among the changed metabolites, cystathionine, l-tyrosine, l-dopa, arginine, alpha-ketoglutaric acid, d-sedoheptulose-7-phosphate and citric acid were significantly decreased in B. bassiana-infected larvae. This indicated that the fungal infection might impair the ability of S. frugiperda larvae to cope with oxidative stress, leading to a negative impact of organism fitness. Further analyses of key metabolic pathways reveal that B. bassiana infection might affect purine metabolism, arginine biosynthesis, butanoate metabolism, and phenylalanine metabolism of S. frugiperda larvae. The findings from this study will contribute to our understanding of oxidative stress on immune defense in insects, and offer fundamental support for the biological control of S. frugiperda.

Different responses of two maize cultivars to Spodoptera frugiperda (Lepidoptera: Noctuidae) larvae infestation provide insights into their differences in resistance.

Spodoptera frugiperda (Lepidoptera: Noctuidae), a pest with an amazing appetite, damages many crops and causes great losses, especially maize. Understanding the differences in different maize cultivars' responses to S. frugiperda infestation is very important for revealing the mechanisms involved in the resistance of maize plants to S. frugiperda. In this study, a comparative analysis of two maize cultivars, the common cultivar 'ZD958' and the sweet cultivar 'JG218', was used to investigate their physico-biochemical responses to S. frugiperda infestation by a pot experiment. The results showed that the enzymatic and non-enzymatic defense responses of maize seedlings were rapidly induced by S. frugiperda. Frist, the hydrogen peroxide (H2O2) and malondialdehyde (MDA) contents of infested maize leaves were significantly increased and then decreased to the level of the control. Furthermore, compared with the control leaves, the puncture force values and the total phenolics, total flavonoids, and 2,4-dihydroxy-7-methoxy-1,4-benzoxazin-3-one contents of infested leaves were significantly increased within a certain time. The superoxide dismutase and peroxidase activities of infested leaves were significantly increased in a certain period of time, while the catalase activities decreased significantly and then increased to the control level. The jasmonic acid (JA) levels of infested leaves were significantly improved, whereas the salicylic acid and abscisic acid levels changed less. Signaling genes associated with phytohormones and defensive substances including PAL4, CHS6, BX12, LOX1, and NCED9 were significantly induced at certain time points, especially LOX1. Most of these parameters changed greater in JG218 than in ZD958. Moreover, the larvae bioassay showed that S. frugiperda larvae weighed more on JG218 leaves than those on ZD958 leaves. These results suggested that JG218 was more susceptible to S. frugiperda than ZD958. Our findings will make it easier to develop strategies for controlling S. frugiperda for sustainable maize production and breeding of new maize cultivars with increased resistance to herbivores.

Physiological and Biochemical Responses in Microalgae Dunaliella salina, Cylindrotheca closterium and Phormidium versicolor NCC466 Exposed to High Salinity and Irradiation.

Dunaliella salina (Chlorophyceae), Phormidium versicolor (Cyanophyceae), and Cylindrotheca closterium (Bacillariophyceae) were isolated from three ponds in the solar saltern of Sfax (Tunisia). Growth, pigment contents, and photosynthetic and antioxidant enzyme activities were measured under controlled conditions of three light levels (300, 500, and 1000 µmol photons m-2 s-1) and three NaCl concentrations (40, 80, and 140 g L-1). The highest salinity reduced the growth of D. salina and P. versicolor NCC466 and strongly inhibited that of C. closterium. According to ΦPSII values, the photosynthetic apparatus of P. versicolor was stimulated by increasing salinity, whereas that of D. salina and C. closterium was decreased by irradiance rise. The production of carotenoids in D. salina and P. versicolor was stimulated when salinity and irradiance increased, whereas it decreased in the diatom. Catalase (CAT), Superoxide dismutase (SOD), and Ascorbate peroxidase (APX) activities were only detected when the three species were cultivated under E1000. The antioxidant activity of carotenoids could compensate for the low antioxidant enzyme activity measured in D. salina. Salinity and irradiation levels interact with the physiology of three species that have mechanisms of more or less effective stress resistance, hence different resistance to environmental stresses according to the species. Under these stress-controlled conditions, P. versicolor and C. closterium strains could provide promising sources of extremolyte for several purposes.

Phytotoxic Action of Silver Nanoparticles on Lemna minor: Multi-Parameter Analysis of Different Physiological Processes.

Considering the widespread use of silver nanoparticles (AgNPs) and their consequent build-up in waterways, there is a concern about the hazardous effect of AgNPs for aquatic ecosystems. The aim of this study was to clarify the mechanism of the action of AgNPs on duckweed (Lemna minor L.) by evaluating multiple parameters in different physiological processes. Duckweed was treated with AgNPs in a concentration range of 0.5 to 5 mg/L over a 7-day period. The analysis revealed that the AgNP-treated duckweed accumulated Ag in accordance with increasing AgNP concentrations. Furthermore, higher concentrations (2 and 5 mg/L) of AgNPs negatively affected N, P and especially K and Mg levels in the plant tissue. Accordingly, the plant growth and photosynthetic parameters were more inhibited in response to higher concentrations of AgNPs. Nanosilver significantly increased the generation of ROS at higher concentrations, although lipid peroxidation was significant even at the lowest concentration of AgNPs. However, defense mechanisms were able to counteract AgNP-induced oxidative stress and balance the intracellular redox status, as evidenced by increased activities of the main detoxification enzymes. With this experimental setting, AgNPs exhibited a relatively weak phytotoxicity at 0.5 and 1 mg/L; nevertheless, silver in a nano form poses a hazard for plants, considering its continuous release into aquatic environments.

Cloning and Function Analysis of ω-3-Fatty Acid Desaturase 8 Gene from Perilla frutescens / 中国生物化学与分子生物学报

ω-3-Fatty acid desaturase 8 (FAD8), as a dehydrogenase enzyme, plays a key role in the transformation of saturated fatty acids into unsaturated fatty acids, which is helpful to enhance the freezing tolerance of plants. However, it remains unclear whether the expression level of FAD8 in Perilla frutescens is regulated by low temperature. Based on transcriptome data, the FAD8 gene was cloned, characterized and then successfully expressed in tobacco Nicotiana tabacum. The gene was designated as PfFAD8 and has a full-length coding sequence of 1 317 bp coding for 438 amino acids with a predicted molecular weight of 50 kD and a theoretical isoelectric point of 9. 13. Our research indicated that the expression of PfFAD8 in Perilla frutescens was increased under the freezing stress. To further confirm this result, a 35S::PfFAD8 vector were constructed and transformed into N. tabacum by Agrobacterium tumefaciens-mediated transformation. Transgenic tobacco leaves that over-expressed the PfFAD8 gene exhibited significantly higher unsaturated fatty acids (UFA) such as linoleic (C18:2) and palmitic acid (C16:0) content and advanced freezing tolerance. Moreover, PfFAD8 overexpression in transgenic tobacco leaves increases malondialdehyde (MDA) and proline (PRO) content, and enhances defense enzymes activities of superoxide dismutase (SOD) and catalase (CAT) to some extent under the cold condition, which might prevent the decline of UFA. Taken together, PfFAD8 overexpression in Perilla frutescens might be involved in the desaturation process of lipids leading to increased membrane stability and/ or induction of other genes related to freezing tolerance by octadecanoid pathway or lipid peroxidation products. Thus, PfFAD8 overexpression could be useful in the production of freeze-tolerant varieties of N. tabacum.

Comparative transcriptomics reveals the immune dynamics during the molting cycle of swimming crab Portunus trituberculatus.

Molting is one of the most important biological processes of crustacean species, and a number of molecular mechanisms facilitate this complex procedure. However, the understanding of the immune mechanisms underlying crustacean molting cycle remains very limited. This study performed transcriptome sequencing in hemolymph and hepatopancreas of the swimming crab (Portunus trituberculatus) during the four molting stages: post-molt (AB), inter-molt (C), pre-molt (D), and ecdysis (E). The results showed that there were 78,572 unigenes that were obtained in the hemolymph and hepatopancreas of P. trituberculatus. Further analysis showed that 98 DEGs were involved in immunity response of hemolymph and hepatopancreas, and most of the DEGs participated in the process of signal transduction, pattern recognition proteins/receptors, and antioxidative enzymes system. Specifically, the key genes and pathway involved in signal transduction including the GPCR126, beta-integrin, integrin, three genes in mitogen-activated protein kinase (MAPK) signaling cascade (MAPKKK10, MAPKK4, and p38 MAPK), and four genes in Toll pathway (Toll-like receptor, cactus, pelle-like kinase, and NFIL3). For the pattern recognition proteins/receptors, the lowest expression level of 11 genes was found in the E stage, including C-type lectin receptor, C-type lectin domain family 6 member A and SRB3/C in the hemolymph, and hepatopancreatic lectin 4, C-type lectin, SRB, Down syndrome cell adhesion molecule homolog, Down syndrome cell adhesion molecule isoform, and A2M. Moreover, the expression level of copper/zinc superoxide dismutase isoform 4, glutathione peroxidase, glutathione S-transferase, peroxiredoxin, peroxiredoxin 6, and dual oxidase 2 in stage C or stage D significantly higher than that of stage E or stage AB. These results fill in the gap of the continuous transcriptional changes that are evident during the molting cycle of crab and further provided valuable information for elucidating the molecular mechanisms of immune regulation during the molting cycle of crab.

Exogenous Cysteine Improves Mercury Uptake and Tolerance in Arabidopsis by Regulating the Expression of Heavy Metal Chelators and Antioxidative Enzymes.

Cysteine (Cys) is an essential amino acid component of the major heavy metal chelators, such as glutathione (GSH), metallothioneins (MTs), and phytochelatins (PCs), which are involved in the pathways of mercury (Hg) tolerance in plants. However, the mechanism through which Cys facilitates Hg tolerance in plants remains largely unclear. In this study, we investigated the effects of exogenous Cys on Hg uptake in the seedlings, roots, and shoots of Arabidopsis throughout 6 and 36 h of Hg exposure and on the regulation of Hg detoxification by heavy metal chelators and antioxidative enzymes. The results showed that exogenous Cys significantly improved Hg tolerance during the germination and seedling growth stages in Arabidopsis. Exogenous Cys significantly promoted Hg uptake in Arabidopsis roots by upregulating the expression of the Cys transporter gene AtLHT1, resulting in increased Hg accumulation in the roots and seedlings. In Arabidopsis seedlings, exogenous Cys further increased the Hg-induced glutathione synthase (GS1 and GS2) transcript levels, and the Hg and Hg + Cys treatments greatly upregulated MT3 expression after 36 h exposure. In the roots, MT3 was also significantly upregulated by treatment of 36 h of Hg or Hg + Cys. Notably, in the shoots, MT2a expression was rapidly induced (10-fold) in Hg presence and further markedly increased (20-fold) by exogenous Cys. Moreover, in the seedlings, exogenous Cys upregulated the transcripts of all superoxide dismutase (CuSOD1, CuSOD2, MnSOD1, FeSOD1, FeSOD2, and FeSOD3) within 6 h and subsequently increased the Hg-induced GR1 and GR2 transcript levels at 36 h, all of which could eliminate the promotion of reactive oxygen species production and cell damage caused by Hg. Additionally, exogenous Cys upregulated all the antioxidative genes rapidly in the roots and subsequently increased the expression of CuSOD1, CuSOD2, and MnSOD1 in the shoots. These results indicate that exogenous Cys regulates the transcript levels of heavy metal chelators and antioxidative enzymes differently in a time- and organ-specific manner under Hg stress. Taken together, our study elucidates the positive functional roles of exogenous Cys in the Hg uptake and tolerance mechanisms of Arabidopsis.

Spraying alginate oligosaccharide improves photosynthetic performance and sugar accumulation in citrus by regulating antioxidant system and related gene expression.

Alginate oligosaccharides (AOS) are functional substances in seaweed extracts that regulate crop quality and stress tolerance. In this paper, the effects of AOS spray application on the antioxidant system, photosynthesis and fruit sugar accumulation in citrus was investigated through a two-year field experiment. The results showed that 8-10 spray cycles of 300-500 mg L-1 AOS (once per 15 days) increased soluble sugar and soluble solid contents by 7.74-15.79% and 9.98-15.35%, respectively, from citrus fruit expansion to harvesting. Compared with the control, the antioxidant enzyme activity and the expression of some related genes in citrus leaves started to increase significantly after the 1st AOS spray application, while the net photosynthetic rate of leaves increased obviously only after the 3rd AOS spray cycle, and the soluble sugar content of AOS-treated leaves increased by 8.43-12.96% at harvest. This suggests that AOS may enhance photosynthesis and sugar accumulation in leaves by antioxidant system regulation. Moreover, analysis of fruit sugar metabolism showed that during the 3rd to 8th AOS spray cycles, AOS treatment increased the activity of enzymes related to sucrose synthesis (SPS, SSs), upregulated the expression of sucrose metabolism (CitSPS1, CitSPS2, SUS) and transport (SUC3, SUC4) genes, and promoted the accumulation of sucrose, glucose and fructose in fruits. Notably, the concentration of soluble sugars in citrus fruits was significantly reduced at all treatments with 40% reduction in leaves of the same branch, but the loss of soluble sugars in AOS-treated fruits (18.18%) was higher than that in the control treatment (14.10%). It showed that there was a positive effect of AOS application on leaf assimilation product transport and fruit sugar accumulation. In summary, AOS application may improve fruit sugar accumulation and quality by regulating the leaf antioxidant system, increasing the photosynthetic rate and assimilate product accumulation, and promoting sugar transfer from leaves to fruits. This study shows the potential application of AOS in the production of citrus fruits for sugar enhancement.

Short-term effects of acute fructose intake on blood antioxidant capacity and liver enzymes in healthy young adults / 中国医师进修杂志

Objective:To investigate the short-term effects of acute fructose intake on serum antioxidant capacity and liver enzymes in healthy young adults.Methods:From January to June 2019, 64 healthy young subjects were recruited, and divided into 75 g glucose group, 25 g fructose group, 50 g fructose group and 75 g fructose group by random digits table method with 16 cases each. The subjects took corresponding amounts of glucose or fructose according to grouping. The levels of alanine aminotransferase (ALT), aspartate aminotransferase (AST), lactate dehydrogenase (LDH), glutathione peroxidase (GPX), superoxide dismutase (SOD), C-Jun amino terminal kinase (JNK), malondialdehyde (MDA) and 8-OH deoxyguanine (8-OHdG) before taking sugar and 30, 60, 120, 180 min after taking sugar, and the changes of ALT, AST and LDH at 30, 60, 120 and 180 min after taking sugar compared with that before taking sugar.Results:One case in 50 g fructose group, 2 cases in 75 g fructose group and 1 case in 75 g glucose group dropped out due to adverse reaction; finally, 15 cases in 75 g glucose group, 16 cases in 25 g fructose group, 15 cases in 50 g fructose group and 14 cases in 75 g fructose group completed the study. The increase of ALT and AST after taking sugar in 25 g fructose group, 50 g fructose group and 75 g fructose group was significantly higher than that in 75 g glucose group, and there were statistical differences ( P<0.05); there was no statistical difference in the change of LDH after taking sugar among 4 groups ( P>0.05). One hundred and eighty min after taking sugar, the receiver operating characteristic (ROC) curve analysis result showed that there were no statistical differences in the areas under curve of ALT, AST and LDH among 4 groups ( P>0.05). There was no statistical difference in SOD before taking sugar among 4 groups ( P>0.05); the SOD 60 min after taking sugar in 50 g fructose group and 75 g fructose group, and SOD 180 min after taking sugar in 25 g fructose group, 50 g fructose group and 75 g fructose group were significantly lower than those in 75 g glucose group: (4.84 ± 1.88) and (4.38 ± 1.12) μg/L vs. (6.25 ± 1.65) μg/L, (4.46 ± 1.66), (5.22 ± 1.66) and (3.99 ± 0.96) μg/L vs. (6.55 ± 1.78) μg/L, and there were statistical differences ( P<0.05). There were no statistical differences in the changes of JNK, GPX, MDA and 8-OHdG before and after taking sugar among 4 groups ( P>0.05). The ROC curve 180 min after taking sugar analysis result showed that the area under curve of SOD in 75 g fructose group was significantly lower than that in 75 g glucose group (9.06 ± 1.88 vs. 12.74 ± 3.15), and there was statistical difference ( P<0.05); there were no statistical differences in the areas under curve of GPX, JNK, MDA and 8-OHdG among 4 groups ( P>0.05). Conclusions:Acute fructose intake can lead to the decrease of antioxidant capacity, and the increasing of oxidative damage and liver enzymes in healthy adults.

Understanding potentially toxic metal (PTM) induced biotic response in two riparian mangrove species Sonneratia caseolaris and Avicennia officinalis along river Hooghly, India: Implications for sustainable sediment quality management.

Elevated human-induced activities have prompted significant uncontrolled release of potentially toxic metals (PTM) to the undisturbed ecosystem throughout the globe. Riparian mangrove vegetations act as a natural purifier of wastewaters and assist in maintaining a healthy ecosystem. We have investigated the elevated PTM-induced stress and biotic response of two riparian mangrove species e.g. Sonneratia caseolaris and Avicennia officinalis by river Hooghly. The increased PTM concentrations were observed throughout the river bank; with the maximum pollution load at Chemaguri (S9). Except Co, Cr and Pb, higher enrichment factor (1.97-8.89) and contamination factor (0.64-2.88) values were observed for Cd, Cu, Fe, Zn. Mn, and Ni. Geo-accumulation index (-2.2 - 0.92) values indicates natural geogenic accumulation of Cu in the riparian mangrove sediment. Thus, sediment quality indices suggest except Cu, enrichment of all studied PTMs was sourced from anthropogenic activities. The sediment of the region when compared with consensus-based sediment quality guidelines shows considerable ecotoxicological risks and threat towards human health considering Ni accumulation. The highest potential ecological risk index value was observed in Chemaguri (S9). The biotic response of riparian mangroves was characterized by reduced photosyhthetic pigments (Chlorophyll a and Chlorophyll b) and increased activity of antioxidative stress enzymes (POD, CAT and SOD). Significant statistical relationship between antioxidative enzyme activity, photosynthetic pigments and bioaccumulated PTMs reflects active functioning of detoxification mechanism in the riparian mangrove species.

The Rapid Cytological Process of Grain Determines Early Maturity in Weedy Rice.

Shorter grain-filling period and rapid endosperm development endow weedy rice (WR) with early maturity compared to cultivated rice (CR). However, the role of the cytological features and antioxidative enzyme system during grain development are largely unexplored. We selected four biotypes of WR and their associated cultivated rice (ACR) types from different latitudes to conduct a common garden experiment. The difference in the cytological features of endosperm between WR and ACR was compared by chemical staining, and the cell viability and nuclear morphometry of endosperm cells were observed by optical microscopy. Furthermore, antioxidative enzyme activity was measured during grain filling. Anatomic observation of endosperm shows that the development process of endosperm cell in WR was more rapid and earlier than that in ACR. The percentage of degraded nuclei of WR was 2-83% more than that of ACR. Endosperm cells in WR lost viability 2-6 days earlier than those in ACR. The antioxidant enzyme activity of WR was lower than that of ACR during grain filling. The ability of WR to scavenge reactive oxygen species (ROS) was weaker than that of ACR, which may contribute to the rapid cytological process in the endosperm cells of WR. The rapid cytological process and weaker ability to scavenge ROS in endosperm cells may contribute to early maturity in WR.

Physiological and Biochemical Response of Wild Olive (Olea europaea Subsp. europaea var. sylvestris) to Salinity.

In the face of climate change, water deficit and increasing soil salinity pose an even greater challenge to olive cultivation in the Mediterranean basin. Due to its tolerance to abiotic stresses, wild olive (Olea europaea subsp. europaea var. sylvestris) presents a good candidate in breeding climate-resilient olive varieties. In this study, the early response of the native Croatian wild olive genotype (WOG) to salinity was evaluated and compared with that of well-known cultivars (cv.) Leccino and Koroneiki. Potted olive plants were exposed either to 150 mM NaCl or 300 mM mannitol for 3 weeks to distinguish between the osmotic and ionic components of salt stress. To determine the impact of the plant age on salinity, 1-, 2-, and 3-year-old WOG plants were used in the study. The growth parameters of both the cultivars and WOG of different ages decreased in response to the mannitol treatment. In contrast to cv. Leccino, the NaCl treatment did not significantly affect the growth of cv. Koroneiki or WOG of any age. The contents of Na+ and Cl- were considerably higher in the salt-treated WOG, regardless of age, compared with the cultivars. However, while both treatments significantly reduced the K+ content of cv. Koroneiki, that nutrient was not significantly affected in either cv. Leccino or WOG. Unlike the cultivars and older WOG, the NaCl treatment caused a significant decline of photosynthetic pigments in the 1-year-old WOG. The cultivars and WOG of different ages experienced a similar drop in the chlorophyll a content under the isotonic mannitol treatment. The absence of lipid peroxidation, modulation of superoxide dismutase, and guaiacol peroxidase activity were noted in all WOG ages under both stressors. These data suggest that WOG resilience to salinity is associated with its large leaf capacity for Na+ and Cl- accumulation, K+ retention, and its adaptable antioxidative mechanisms. The results are promising with regard to obtaining a new olive cultivar with better resilience to soil salinity.

Effect of drought on photosynthesis, total antioxidant capacity, bioactive component accumulation, and the transcriptome of Atractylodes lancea.

BACKGROUND: Atractylodes lancea (Thunb.) DC, a medicinal herb belonging to the Asteraceae family, often faces severe drought stress during its growth. Until now, there has been no research on the effect of drought stress on the quality formation of A. lancea. Therefore, the present study aimed to study the effects of drought stress on A. lancea through physical and chemical analysis, and to reveal the related molecular mechanisms via transcriptome analysis. RESULTS: The photosynthesis was markedly inhibited under drought stress. There were alterations to photosynthetic parameters (Pn, Gs, Ci) and chlorophyll fluorescence (Fv/Fm, NPQ), and the chlorophyll content decreased. Twenty genes encoding important regulatory enzymes in light and dark reactions, including the Rubisco gene of the Calvin cycle, were significantly downregulated. After exposure to drought stress for more than 4 days, the activities of four antioxidative enzymes (SOD, POD CAT and APX) began to decrease and continued to decrease with longer stress exposure. Meanwhile, most of the genes encoding antioxidative enzymes were downregulated significantly. The downregulation of 21 genes related to the respiratory electron transport chain indicated that the blocked electron transfer accelerated excessive ROS. The MDA content was significantly elevated. The above data showed that 15 days of drought stress caused serious oxidative damage to A. lancea. Drought stress not only reduced the size and dry weight of A. lancea, but also lowered the amount of total volatile oil and the content of the main bioactive components. The total volatile oil and atractylodin content decreased slightly, whereas the content of atractylon and ß-eudesmol decreased significantly. Moreover, ten significantly downregulated genes encoding sesquiterpene synthase were mainly expressed in rhizomes. CONCLUSIONS: After exposed to drought stress, the process of assimilation was affected by the destruction of photosynthesis; stress tolerance was impaired because of the inhibition of the antioxidative enzyme system; and bioactive component biosynthesis was hindered by the downregulation of sesquiterpene synthase-related gene expression. All these had negative impacts on the quality formation of A. lancea under drought stress.

Intense Pulsed Light Attenuates UV-Induced Hyperimmune Response and Pigmentation in Human Skin Cells.

The skin of an organism is affected by various environmental factors and fights against aging stress via mechanical and biochemical responses. Photoaging induced by ultraviolet B (UVB) irradiation is common and is the most vital factor in the senescence phenotype of skin, and so, suppression of UVB stress-induced damage is critical. To lessen the UVB-induced hyperimmune response and hyperpigmentation, we investigated the ameliorative effects of intense pulsed light (IPL) treatment on the photoaged phenotype of skin cells. Normal human epidermal keratinocytes and human epidermal melanocytes were exposed to 20 mJ/cm2 of UVB. After UVB irradiation, the cells were treated with green (525-530 nm) and yellow (585-592 nm) IPL at various time points prior to the harvest step. Subsequently, various signs of excessive immune response, including expression of proinflammatory and melanogenic genes and proteins, cellular oxidative stress level, and antioxidative enzyme activity, were examined. We found that IPL treatment reduced excessive cutaneous immune reactions by suppressing UVB-induced proinflammatory cytokine expression. IPL treatment prevented hyperpigmentation, and combined treatment with green and yellow IPL synergistically attenuated both processes. IPL treatment may exert protective effects against UVB injury in skin cells by attenuating inflammatory cytokine and melanogenic gene overexpression, possibly by reducing intracellular oxidative stress. IPL treatment also preserves antioxidative enzyme activity under UVB irradiation. This study suggests that IPL treatment is a useful strategy against photoaging, and provides evidence supporting clinical approaches with non-invasive light therapy.

Comparative response of Spodoptera litura challenged per os with Serratia marcescens strains differing in virulence.

Host plays an important role in influencing virulence of a pathogen and efficacy of a biopesticide. The present study was aimed to characterize the possible factors present in Spodoptera litura that influenced pathogenecity of orally ingested S. marcescens strains, differing in their virulence. Fifth instar larvae of S. litura responded differently as challenged by two Serratia marcescens strains, SEN (virulent strain, LC50 7.02 103 cfu/ml) and ICC-4 (non-virulent strain, LC50 1.19 1012 cfu/ml). Considerable increase in activity of lytic enzymes protease and phospholipase was recorded in the gut and hemolymph of larvae fed on diet supplemented with S. marcescens strain ICC-4 as compared to the larvae treated with S. marcescens strain SEN. However, a significant up-regulation of antioxidative enzymes SOD (in foregut and midgut), CAT (in the midgut) and GST (in the foregut and hemolymph) was recorded in larvae fed on diet treated with the virulent S. marcescens strain SEN in comparison to larvae fed on diet treated with the non-virulent S. marcescens strain ICC-4. Activity of defense related enzymes lysozyme and phenoloxidase activity were also higher in the hemolymph of larvae fed with diet treated with S. marcescens strain SEN as compared to hemolymph of S. marcescens strain ICC-4 treated larvae. More number of over-expressed proteins was observed in the gut and hemolymph of S. marcescens strains ICC-4 and SEN treated larvae, respectively. Identification of the selected differentially expressed proteins indicated induction of proteins involved in insect innate immune response (Immunoglobulin I-set domain, Apolipophorin III, leucine rich repeat and Titin) in S. marcescens strain SEN treated larvae. Over-expression of two proteins, actin related protein and mt DNA helicase, were noted in S. marcescens treated larvae with very high levels observed in the non-virulent strain. Up-regulation of homeobox protein was noted only in S. marcescens strain ICC-4 challenged larvae. This study indicated that ingestion of non-virulent S. marcescens strain ICC-4 induced strong immune response in insect gut while there was weak response to the virulent S. marcescens strain SEN which probably resulted in difference in their virulence.

The Influence of Soil Acidity on the Physiological Responses of Two Bread Wheat Cultivars.

The recent study was conducted to examine the influence of acidic soil on the activities of ascorbate (APX) and guaiacol peroxidase (POD), proline, protein as well as malon-dialdehyde (MDA) content, in two commercial spring wheat cultivars (PAN3497 and SST806) at different growth stages (tillering and grain filling). A cultivar effect was significant only for MDA content, while the treatment effect was highly significant for proline, protein, and MDA. The sampling time effect was significant for most characteristics. MDA, antioxidative capacity, as well as protein content increased with maturity. At grain filling, MDA and proline contents were significantly higher at pH 5 than pH 6 and 7 for both cultivars, with the highest content in SST806. Similarly, SST806 had significantly higher APX and POD when growing at pH 5. There were no significant differences in protein content at grain filling between either genotype or treatments affected by low pH. This study showed that growth stage and soil pH influence the rate of lipid peroxidation as well as the antioxidative capacity of wheat, with a larger effect at grain filling, at pH 5. Although SST806 had higher proline, POD, and APX content than PAN3497 at this growth stage, this coincided with a very high MDA content. This shows that the high antioxidative capacity observed here, was not associated with a reduction of lipid peroxidation under low soil pH. Further research should, therefore, be done to establish the role of the induced antioxidant system in association with growth and yield in wheat.