HEAT-RESPONSIVE PROTEIN regulates heat stress via fine-tuning ethylene/auxin signaling pathways in cotton.

Plants sense and respond to fluctuating temperature and light conditions during the circadian cycle; however, the molecular mechanism underlying plant adaptability during daytime warm conditions remains poorly understood. In this study, we reveal that the ectopic regulation of a HEAT RESPONSIVE PROTEIN (GhHRP) controls the adaptation and survival of cotton (Gossypium hirsutum) plants in response to warm conditions via modulating phytohormone signaling. Increased ambient temperature promptly enhanced the binding of the phytochrome interacting factor 4 (GhPIF4)/ethylene-insensitive 3 (GhEIN3) complex to the GhHRP promoter to increase its mRNA level. The ectopic expression of GhHRP promoted the temperature-dependent accumulation of GhPIF4 transcripts and hypocotyl elongation by triggering thermoresponsive growth-related genes. Notably, the upregulation of the GhHRP/GhPIF4 complex improved plant growth via modulating the abundance of Arabidopsis thaliana auxin biosynthetic gene YUCCA8 (AtYUC8)/1-aminocyclopropane-1-carboxylate synthase 8 (AtACS8) for fine-tuning the auxin/ethylene interplay, ultimately resulting in decreased ethylene biosynthesis. GhHRP thus protects chloroplasts from photo-oxidative bursts via repressing AtACS8 and AtACS7 and upregulating AtYUC8 and the heat shock transcription factors (HSFA2), heat shock proteins (HSP70 and HSP20). Strikingly, the Δhrp disruption mutant exhibited compromised production of HSP/YUC8 that resulted in an opposite phenotype with the loss of the ability to respond to warm conditions. Our results show that GhHRP is a heat-responsive signaling component that assists plants in confronting the dark phase and modulates auxin signaling to rescue growth under temperature fluctuations.

Twelve tips for Natural Language Processing in medical education program evaluation.

With the increasing application of Natural Language Processing (NLP) in Medicine at large, medical educators are urged to gain an understanding and implement NLP techniques within their own education programs to improve the workflow and make significant and rapid improvements in their programs. This paper aims to provide twelve essential tips inclusive of both conceptual and technical factors to facilitate the successful integration of NLP in medical education program evaluation. These twelve tips range from advising on various stages of planning the evaluation process, considerations for data collection, and reflections on preprocessing of data in preparation for analysis and interpretation of results. Using these twelve tips as a framework, medical researchers, educators, and administrators will have an understanding and reference to navigating applications of NLP and be able to unlock its potential for enhancing the evaluation of their own medical education programs.

Biotite: new tools for a versatile Python bioinformatics library.

BACKGROUND: Biotite is a program library for sequence and structural bioinformatics written for the Python programming language. It implements widely used computational methods into a consistent and accessible package. This allows for easy combination of various data analysis, modeling and simulation methods. RESULTS: This article presents major functionalities introduced into Biotite since its original publication. The fields of application are shown using concrete examples. We show that the computational performance of Biotite for bioinformatics tasks is comparable to individual, special purpose software systems specifically developed for the respective single task. CONCLUSIONS: The results show that Biotite can be used as program library to either answer specific bioinformatics questions and simultaneously allow the user to write entire, self-contained software applications with sufficient performance for general application.

The Interplay between Hydrogen Sulfide and Phytohormone Signaling Pathways under Challenging Environments.

Hydrogen sulfide (H2S) serves as an important gaseous signaling molecule that is involved in intra- and intercellular signal transduction in plant-environment interactions. In plants, H2S is formed in sulfate/cysteine reduction pathways. The activation of endogenous H2S and its exogenous application has been found to be highly effective in ameliorating a wide variety of stress conditions in plants. The H2S interferes with the cellular redox regulatory network and prevents the degradation of proteins from oxidative stress via post-translational modifications (PTMs). H2S-mediated persulfidation allows the rapid response of proteins in signaling networks to environmental stimuli. In addition, regulatory crosstalk of H2S with other gaseous signals and plant growth regulators enable the activation of multiple signaling cascades that drive cellular adaptation. In this review, we summarize and discuss the current understanding of the molecular mechanisms of H2S-induced cellular adjustments and the interactions between H2S and various signaling pathways in plants, emphasizing the recent progress in our understanding of the effects of H2S on the PTMs of proteins. We also discuss future directions that would advance our understanding of H2S interactions to ultimately mitigate the impacts of environmental stresses in the plants.

Challenges and prospects for a potential allohexaploid Brassica crop.

The production of a new allohexaploid Brassica crop (2n = AABBCC) is increasingly attracting international interest: a new allohexaploid crop could benefit from several major advantages over the existing Brassica diploid and allotetraploid species, combining genetic diversity and traits from all six crop species with additional allelic heterosis from the extra genome. Although early attempts to produce allohexaploids showed mixed results, recent technological and conceptual advances have provided promising leads to follow. However, there are still major challenges which exist before this new crop type can be realized: (1) incorporation of sufficient genetic diversity to form a basis for breeding and improvement of this potential crop species; (2) restoration of regular meiosis, as most allohexaploids are genetically unstable after formation; and (3) improvement of agronomic traits to the level of "elite" breeding material in the diploid and allotetraploid crop species. In this review, we outline these major prospects and challenges and propose possible plans to produce a stable, diverse and agronomically viable allohexaploid Brassica crop.

Drought tolerance in Brassica napus is accompanied with enhanced antioxidative protection, photosynthetic and hormonal regulation at seedling stage.

Climate change, food insecurity, water scarcity, and population growth are some of today's world's frightening problems. Drought stress exerts a constant threat to field crops and is often seen as a major constraint on global agricultural productivity; its intensity and frequency are expected to increase in the near future. The present study investigated the effects of drought stress (15% w/v polyethylene glycol PEG-6000) on physiological and biochemical changes in five Brassica napus cultivars (ZD630, ZD622, ZD619, GY605, and ZS11). For drought stress induction, 3-week-old rapeseed oil seedlings were treated with PEG-6000 in full strength Hoagland nutrient solution for 7 days. PEG treatment significantly decreased the plant growth and photosynthetic efficiency, including primary photochemistry (Fv/Fm) of PSII, intercellular CO2 , net photosynthesis, chlorophyll contents, and water-use efficiency of all studied B. napus cultivars; however, pronounced growth retardations were observed in cultivar GY605. Drought-stressed B. napus cultivars also experienced a sharp rise in H2 O2 generation and malondialdehyde (MDA) content. Additionally, the accumulation of ROS was accompanied by increased activity of enzymatic antioxidants (superoxide dismutase, peroxidase, catalase, ascorbate peroxidase, glutathione reductase, and monodehydroascorbate reductase), although the increase was more obvious in ZD622 and ZS11. Drought stress also caused an increased endogenous hormonal biosynthesis (abscisic acid, jasmonic acid, salicylic acid) and accumulation of total soluble proteins and proline content, but the extent varies in B. napus cultivars. These results suggest that B. napus cultivars have an efficient drought stress tolerance mechanism, as shown by improved antioxidant enzyme activities, photosynthetic and hormonal regulation.

Comprehensive proteomic analysis of arsenic induced toxicity reveals the mechanism of multilevel coordination of efficient defense and energy metabolism in two Brassica napus cultivars.

Arsenic (As) a non-essential element is of particular concern with respect to harmful effects on plant metabolism. While extensive studies have been conducted on the physiological responses of plants to increase As concentrations, however, molecular differences elucidating species-specific changes remain largely unknown. In the present experiment, two oilseed Brassica napus (B. napus) cultivars, ZS758 and ZD622, were treated by elevated As concentration. Their responses to the As stress have been investigated through pulse amplitude modulated fluorometer and isobaric tags based proteomic (iTRAQ) analysis. The chlorophyll fluorescence attributes showed that As stress significantly decrease the photochemical efficiency of photosystem II (PSII) and photosystem I (PSI) as well as the comparatively closed stomata observed under scanning electron microscopy (SEM). In this study, 65 proteins displayed increased abundance and 52 down-regulated were found in the control vs As comparison in cultivar ZS758, while 44 up and 67 down-regulated proteins were found in the control vs As comparison in ZD622. Metabolic pathways, followed by ribosome and biosynthesis of secondary metabolites were the dominant functional annotation categories among the differentially expressed protein (DEPs). Many genes involved in primary metabolism, stress and defense were found to be As-responsive DEPs and/or DEPs between these two cultivars. Based on these results, a schematic description of key processes involved in As tolerance in ZS758 and ZD622 is proposed, which suggests that higher tolerance in ZS758 depends on a multilevel coordination of efficient defense and energy metabolism. Real-time quantitative PCR supported the expression patterns of several genes encoding a protein similar to their corresponding DEPs. In addition, these findings could shed light in unraveling the molecular mechanisms of B. napus exposed to As stress and provide or improve essential understandings in the development of advanced B. napus cultivars against As resistance.

The Effect of Virulence and Resistance Mechanisms on the Interactions between Parasitic Plants and Their Hosts.

Parasitic plants have a unique heterotrophic lifestyle based on the extraction of water and nutrients from host plants. Some parasitic plant species, particularly those of the family Orobanchaceae, attack crops and cause substantial yield losses. The breeding of resistant crop varieties is an inexpensive way to control parasitic weeds, but often does not provide a long-lasting solution because the parasites rapidly evolve to overcome resistance. Understanding mechanisms underlying naturally occurring parasitic plant resistance is of great interest and could help to develop methods to control parasitic plants. In this review, we describe the virulence mechanisms of parasitic plants and resistance mechanisms in their hosts, focusing on obligate root parasites of the genera Orobanche and Striga. We noticed that the resistance (R) genes in the host genome often encode proteins with nucleotide-binding and leucine-rich repeat domains (NLR proteins), hence we proposed a mechanism by which host plants use NLR proteins to activate downstream resistance gene expression. We speculated how parasitic plants and their hosts co-evolved and discussed what drives the evolution of virulence effectors in parasitic plants by considering concepts from similar studies of plant-microbe interaction. Most previous studies have focused on the host rather than the parasite, so we also provided an updated summary of genomic resources for parasitic plants and parasitic genes for further research to test our hypotheses. Finally, we discussed new approaches such as CRISPR/Cas9-mediated genome editing and RNAi silencing that can provide deeper insight into the intriguing life cycle of parasitic plants and could potentially contribute to the development of novel strategies for controlling parasitic weeds, thereby enhancing crop productivity and food security globally.

Comparative Transcriptomic Analysis of Biological Process and Key Pathway in Three Cotton (Gossypium spp.) Species Under Drought Stress.

Drought is one of the most important abiotic stresses that seriously affects cotton growth, development, and production worldwide. However, the molecular mechanism, key pathway, and responsible genes for drought tolerance incotton have not been stated clearly. In this research, high-throughput next generation sequencing technique was utilized to investigate gene expression profiles of three cotton species (Gossypium hirsutum, Gossypium arboreum, and Gossypium barbadense L.) under drought stress. A total of 6968 differentially expressed genes (DEGs) were identified, where 2053, 742, and 4173 genes were tested as statistically significant; 648, 320, and 1998 genes were up-regulated, and 1405, 422, and 2175 were down-regulated in TM-1, Zhongmian-16, and Pima4-S, respectively. Total DEGs were annotated and classified into functional groups under gene ontology analysis. The biological process was present only in tolerant species(TM-1), indicating drought tolerance condition. The Kyoto encyclopedia of genes and genomes showed the involvement of plant hormone signal transduction and metabolic pathways enrichment under drought stress. Several transcription factors associated with ethylene-responsive genes (ICE1, MYB44, FAMA, etc.) were identified as playing key roles in acclimatizing to drought stress. Drought also caused significant changes in the expression of certain functional genes linked to abscisic acid (ABA) responses (NCED, PYL, PP2C, and SRK2E), reactive oxygen species (ROS) related in small heat shock protein and 18.1 kDa I heat shock protein, YLS3, and ODORANT1 genes. These results will provide deeper insights into the molecular mechanisms of drought stress adaptation in cotton.

Physiological and iTRAQ-Based Quantitative Proteomics Analysis of Methyl Jasmonate-Induced Tolerance in Brassica napus Under Arsenic Stress.

Brassica napus plants exposed to 200 µM arsenic (As) exhibited high-level of stress condition, which led to inhibited growth, enhanced lipid peroxidation, and disrupted cellular ultrastructures. Exogenous application of methyl jasmonate (MeJA) alleviated the As-induced oxidative stress and improved the plant growth and photosynthesis. In this study, changes in the B. napus leaf proteome are investigated in order to identify molecular mechanisms involved in MeJA-induced As tolerance. The study identifies 177 proteins that are differentially expressed in cultivar ZS 758; while 200 differentially expressed proteins are accumulated in Zheda 622, when exposed to As alone and MeJA+As treatments, respectively. The main objective was to identify the MeJA-regulated protein under As stress. Consistent with this, iTRAQ detected 61 proteins which are significantly accumulated in ZS 758 leaves treated with MeJA under As stress. While in Zheda 622, iTRAQ detected 49 MeJA-induced proteins under As stress. These significantly expressed proteins are further divided into five groups on the base of their function, that is, stress and defense, photosynthesis, carbohydrates and energy production, protein metabolism, and secondary metabolites. Taken together, this study sheds light on the molecular mechanisms involved in MeJA-induced As tolerance in B. napus leaves and suggests a more active involvement of MeJA in plant physiological processes.

iTRAQ-based proteomics of sunflower cultivars differing in resistance to parasitic weed Orobanche cumana.

Orobanche cumana is an obligate root parasite causing severe damage to many economically important crops, including sunflowers worldwide. For efficient control measures, it is necessary to understand the resistant mechanism during interaction at molecular level. The present study emphasizes on comparative proteomics to investigate the mechanistic basis of compatible and incompatible interaction of O. cumana with resistant (JY207) and susceptible (TK0409) sunflowers. More than 3500 proteins were identified from two cultivars by iTRAQ analysis. Identified proteins associated with general functions, posttranslational modification, energy production and conversion, carbohydrate transport and metabolism, and signal transduction mechanisms were the most represented category of induced proteins in both cultivars. The resistant interaction was characterized by alteration of defense-related proteins involved in recognition of parasites, accumulation of pathogenesis-related proteins, biosynthesis of lignin, and detoxification of toxic metabolites in JY207 after inoculation. The susceptible interaction was characterized by decreased abundance of proteins involved in biosynthesis and signaling of plant growth regulators including auxin, gibberellin, brassinosteroid, and ethylene in TK0409 after inoculation. The present study provides comprehensive details of proteins and differential modulation of pathways regulated under compatible and incompatible interaction, allowing the identification of important molecular components for development of sustainable resistance against this parasite.

Biochemical responses and ultrastructural changes in ethylene insensitive mutants of Arabidopsis thialiana subjected to bisphenol A exposure.

Bisphenol A (BPA), an important raw material in plastic industry, has become a serious environmental contaminant due to its wide spread use in different products and increasing release into the environment. BPA is known to cause adverse effects in living organisms including plants. Several studies reported that BPA affects growth and development in plants, mainly through oxidative stress. Plants are known to generally cope with stress mainly through hormonal regulation and adaptation, but little is known about the role of plant hormones in plants under BPA stress. The present study was conducted to investigate the role of ethylene in BPA induced oxidative stress in plants using Arabidopsis thaliana as a test plant. The response of ethylene insensitive mutants of Arabidopsis (ein2-1 and etr1-3) to BPA exposure was studied in comparison to the wild type Arabidopsis (WT). In all three genotypes, exposure to BPA adversely affected cellular structures, stomata and light-harvesting pigments. An increase in reactive oxygen species (ROS) lipid peroxidation and other oxidative stress markers indicated that BPA induced toxicity through oxidative stress. However, the overall results revealed that WT Arabidopsis had more pronounced BPA induced damages while ein2-1 and etr1-3 mutants withstood the BPA induced stress more efficiently. The activity of antioxidant enzymes and expression of antioxidants related genes revealed that the antioxidant defense system in both mutants was more efficiently activated than in WT against BPA induced oxidative stress, which further evidenced the involvement of ethylene in regulating BPA induced oxidative stress. It is concluded that ethylene perception and signaling may be involved in BPA induced oxidative stress responses in plants.

Complementary RNA-Sequencing Based Transcriptomics and iTRAQ Proteomics Reveal the Mechanism of the Alleviation of Quinclorac Stress by Salicylic Acid in Oryza sativa ssp. japonica.

To uncover the alleviation mechanism of quinclorac stress by salicylic acid (SA), leaf samples of Oryza sativa ssp. Japonica under quinclorac stress with and without SA pre-treatment were analyzed for transcriptional and proteomic profiling to determine the differentially expressed genes (DEGs) and proteins (DEPs), respectively. Results showed that quinclorac stress altered the expression of 2207 DEGs (1427 up-regulated, 780 down-regulated) and 147 DEPs (98 down-regulated, 49 up-regulated). These genes and proteins were enriched in glutathione (GSH) metabolism, porphyrin and chlorophyll metabolism, the biosynthesis of secondary metabolites, glyoxylate and dicarboxylate metabolism, and so on. It also influenced apetala2- ethylene-responsive element binding protein (AP2-EREBP) family, myeloblastosis (MYB) family and WRKY family transcription factors. After SA pre-treatment, 697 genes and 124 proteins were differentially expressed. Pathway analysis showed similar enrichments in GSH, glyoxylate and dicarboxylate metabolism. Transcription factors were distributed in basic helix-loop-helix (bHLH), MYB, Tify and WRKY families. Quantitative real-time PCR results revealed that quinclorac stress induced the expression of glutathion reductase (GR) genes (OsGR2, OsGR3), which was further pronounced by SA pre-treatment. Quinclorac stress further mediated the accumulation of acetaldehyde in rice, while SA enhanced the expression of OsALDH2B5 and OsALDH7 to accelerate the metabolism of herbicide quinclorac for the protection of rice. Correlation analysis between transcriptome and proteomics demonstrated that, under quinclorac stress, correlated proteins/genes were mainly involved in the inhibition of intermediate steps in the biosynthesis of chlorophyll. Other interesting proteins/genes and pathways regulated by herbicide quinclorac and modulated by SA pre-treatment were also discussed, based on the transcriptome and proteomics results.

Comparative transcriptome profiling of two Brassica napus cultivars under chromium toxicity and its alleviation by reduced glutathione.

BACKGROUND: Chromium (Cr) being multifarious industrial used element, is considered a potential environmental threat. Cr found to be a prospective water and soil pollutant, and thus it is a current area of concern. Oilseed rape (Brassica napus L.) is well known as a major source of edible oil around the globe. Due to its higher growth, larger biomass and capability to uptake toxic materials B. napus is considered a potential candidate plant against unfavorable conditions. To date, no study has been done that described the Cr and GSH mechanism at RNA-Seq level. RESULTS: Both digital gene expression (DGE) and transcriptome profile analysis (TPA) approaches had opened new insights to uncover the several number of genes related to Cr stress and GSH alleviating mechanism in two leading cultivars (ZS 758 and Zheda 622) of B. napus plants. Data showed that Cr inhibited KEGG pathways i.e. stilbenoid, diarlyheptanoid and gingerol biosynthesis; limonene and pentose degradation and glutathione metabolism in ZS 758; and ribosome and glucosinolate biosynthesis in Zheda-622. On the other hand, vitamin B6, tryptophan, sulfur, nitrogen and fructose and manose metabolisms were induced in ZS 758, and zeatin biosynthesis, linoleic acid metabolism, arginine and proline metabolism, and alanine, asparate and glutamate metabolism pathways in Zheda 622. Cr increased the TFs that were related to hydralase activity, antioxidant activity, catalytic activity phosphatase and pyrophosphatase activity in ZS 758, and vitamin binding and oxidoreductase activity in Zheda 622. Cr also up-regulated the promising proteins related to intracellular membrane bounded organelles, nitrile hyrdatase activity, cytoskeleton protein binding and stress response. It also uncovered, a novel Cr-responsive protein (CL2535.Contig1_All) that was statistically increased as compared to control and GSH treated plants. Exogenously applied GSH successfully not only recovered the changes in metabolic pathways but also induced cysteine and methionine metabolism in ZS 758 and ubiquinone and other terpenoid-quinone biosynthesis pathways in Zheda 622. Furthermore, GSH increased the level of TFs i.e. the gene expression of antioxidant and catalytic activities, iron ion binding and hydrolase activity as compared with Cr. Moreover, results pointed out a novel GSH responsive protein (CL827.Contig3_All) whose expression was found to be significantly increased when compared than Cr stress. Results further delineated that GSH induced TFs such as glutathione disulphide oxidoreducatse and aminoacyl-tRNA ligase activity, and beta glucosidase activity in ZS 758. Similarly in Zheda 622, GSH induced the TFs for instance DNA binding and protein dimerization activity. GSH also highlighted the proteins that were involved in transportation, photosynthesis process, RNA polymerase activity, and against the metal toxicity. These results indicated that cultivar ZS 758 had better metabolism and showed higher tolerance against Cr toxicity. CONCLUSION: The responses of ZS 758 and Zheda 622 differed considerably at both physiological and transcriptional level. Moreover, RNA-Seq method explored the hazardous behavior of Cr as well as GSH up-regulating mechanism by activating plant metabolism, stress responsive genes, TFs and protein encyclopedia.

Salicylic acid mediates antioxidant defense system and ABA pathway related gene expression in Oryza sativa against quinclorac toxicity.

The auxin herbicide quinclorac is widely used for controlling weeds in transplanted and direct-seeded rice fields. However, its phytotoxic responses on rice are still unknown. Therefore, in the present investigation we studied the effects of different concentrations (0, 0.1 and 0.5g/L) of quinclorac herbicide on the physiological and biochemical changes of two rice cultivars (XS 134 and ZJ 88) and further analyzed the ameliorating role of salicylic acid (SA) on quinclorac toxicity in rice plants. The results revealed that exogenous application of SA significantly increased plant biomass and total chlorophyll contents in herbicide stressed plants. The lipid peroxidation and ROS (H2O2, O2(-.), (-)OH) production were significantly increased in roots and leaves of both rice cultivars under quinclorac stress, demonstrating an oxidative burst in rice plants. Whereas, application of SA significantly lowered ROS contents under quinclorac stress. Further, exogenous SA treatment significantly modulated antioxidant enzymes and enhanced GSH concentration in stress plants. Anatomical observations of leaf and root revealed that herbicide affected internal structures, while SA played a vital role in protection from toxic effects. Expression analysis of stress hormone ABA genes (OsABA8oxs, OsNCEDs) revealed that quinclorac application enhanced stress condition in cultivar ZJ 88, while SA treatment downregulated ABA genes more in cultivar XS 134, which correlated with the enhanced tolerance to quinclorac induced oxidative stress in this cultivar. The present study delineated that SA played a critical role under quinclorac stress in both rice cultivars by regulating antioxidant defense system, reducing ROS formation and preventing the degradation of internal cell organelles.

Toxicological effects of bisphenol A on growth and antioxidant defense system in Oryza sativa as revealed by ultrastructure analysis.

The present study was conducted to evaluate the effect of bisphenol A (BPA) on rice seedlings grown in a hydroponic system. The obtained results demonstrated that at lower concentration (up to 10µM) BPA had some stimulatory effects on the growth of rice seedlings but at higher doses it significantly inhibited seedlings growth. The photosynthetic pigments were significantly decreased by high doses of BPA. Exposure to BPA caused increased membrane permeability in root cells and resulted in increased concentration of reactive oxygen species (ROS) and increased lipid peroxidation as revealed by thiobarbituric acid reactive substances (TBARS) assay. In leaves, superoxide dismutase (SOD) activity exhibited an increase at lower concentrations of BPA but was inhibited at the highest dose (200µM) of BPA. At 100µM of BPA, a significant increase in antioxidant activities in leaves was observed but at 200µM this activity was inhibited. In roots, a significant decrease in enzymes activity was recorded at the highest concentration of BPA (200µM); however, ascorbate peroxidase (APX) and catalase (CAT) activities were significantly increased at the concentrations of 10 and 50µM in comparison to the control. Moreover, the present results revealed that BPA severely affected cell organelles in rice seedlings. It can be concluded that the observed adverse effects in rice seedlings by BPA in the present study could be attributed to the oxidative stress caused by BPA.

Differential subcellular distribution and chemical forms of cadmium and copper in Brassica napus.

Metal subcellular fractions and chemical profile highly reflect their level of toxicity to plants. Cadmium and Cu, two different but potentially toxic metals, were compared in the present study for their subcellular distribution and chemical forms in two Brassica napus cultivars (Zheda 622 and ZS 758). Five-week-old seedlings were hydroponically exposed to metal stress and analyzed after 15 days of treatment. In both cultivars, Cd was less retained at cell wall, thus major part of Cd accumulated in the soluble fraction. By contrast, handsome amount of Cu was sequestrated in both cell wall and vacuole containing fraction. Across sensitive organelles, Cu preferentially accumulated in chloroplasts, while Cd was equally distributed in chloroplasts and mitochondria; the two metals intruded nucleus at lesser degree. Further, Cd and Cu differentially interacted with various cellular ligands, and the extent of interaction was higher in the tolerant cultivar ZS 758. Copper was remarkably sequestrated by phosphates, and secondarily by peptide-ligands; inversely, the role of phosphates was secondary in Cd complexation, which was mainly achieved by peptide-ligands. Additional amount of Cu was aggregated with oxalates, but oxalate-bound Cd was scarcely detected. Current results have demonstrated varied toxicological and detoxification pathways of Cd and Cu in B. napus, suggesting that the efficiency of different alleviation strategies could vary against Cd and Cu toxicity to plants.

Subcellular distribution, modulation of antioxidant and stress-related genes response to arsenic in Brassica napus L.

Arsenic (As) is an environmental toxin pollutant that affects the numerous physiological processes of plants. In present study, two Brassica napus L. cultivars were subjected to various concentrations (0, 50, 100, and 200 µM) of As for 14 days, plants were examined for As subcellular distribution, photosynthesis parameters, oxidative stress, and ultrastructural changes under As-stress. Differential fraction analysis showed that significant amount of As was accumulated in the cell wall as compared to other organelles. Decline in photosynthetic efficiency under As stress was observed in term of reduced pigment contents and gas exchange parameters. Differential responses of antioxidants at both enzymatic and gene levels to higher As stress were more pronounced in cultivar ZS 758 as compared to Zheda 622. The qRT-PCR analysis showed that heat shock protein 90 (Hsp90) and metallothionein were over-expressed in As stressed B. napus plants. Disorganization of cell structure and the damages in different organelles were some of the obvious variations in cultivar Zheda 622 as compared to ZS 758.

Synergism of herbicide toxicity by 5-aminolevulinic acid is related to physiological and ultra-structural disorders in crickweed (Malachium aquaticum L.).

Selection of effective herbicides to control weeds has been one of the major objectives of scientists. This study determines the differential tolerance or susceptibility of crickweed (Malachium aquaticum L.) to various concentration combinations of 5-aminolevulinic acid (ALA) (1, 10 and 100mg/L) and propyl 4-(2-(4,6-dimethoxypyrimidin-2-yloxy)benzylamino)benzoate (ZJ0273) (100, 200, and 500mg/L). ALA was applied as pre- and post-treatment alone or in combination with ZJ0273. Results showed that ZJ0273 stress alone imposed negative effects on M. aquaticum seedling's growth, net photosynthetic rates and SPAD values, and the rate of decline was consistently increased with the increase in ZJ0273 concentration. The ZJ0273 treatment showed a gradual decrease in the activities of antioxidant enzymes peroxidase (POD), superoxide dismutase (SOD), and ascorbate peroxidase (APX), and increase in the accumulation of malondialdehyde (MDA). Changes in chloroplast swelling, increased number of plastoglobuli, disruption of thylakoid, disintegrated mitochondria and turbid nucleoplasm were noticed. Moreover, SDS-PAGE analysis of total proteins revealed that herbicide stress in the leaves was associated with the decrease or disappearance of some protein bands. Further, two-dimensional gel electrophoresis (2-DE) results showed that proteins in different spots were classified into three types for M. aquaticum. These results indicate that the combined treatment of ALA and ZJ0273 synergizes the herbicide toxicity which is different from its independent effects on M. aquaticum and thus, could improve weed control efficacy.