Main Molecular Pathways Associated with Copper Tolerance Response in Imperata cylindrica by de novo Transcriptome Assembly

The metallophyte Imperata cylindrica inhabits copper (Cu) polluted soils in large areas from Central Chile. Here, we subjected clonal vegetative plantlets to 300 mg Cu kg−1 of substrate for 21 days to identify the main molecular pathways involved in the response to Cu stress. Transcriptomic analyses were performed for shoots and roots, with and without Cu supply. RNA-Seq and de novo transcriptome assembly were performed to identify the gene response associated with molecular mechanisms of Cu tolerance in I. cylindrica. De novo transcriptome revealed a total of 200,521 transcripts (1777 bp) comprising ~91% complete ultra-conserved genes in the eukaryote and Plantae database. The differentially expressed genes (DEGs) in roots were 7386, with 3558 of them being up-regulated and the other 3828 down-regulated. The transcriptome response in shoots was significantly less, showing only 13 up-regulated and 23 down-regulated genes. Interestingly, DEGs mainly related with actin and cytoskeleton formation, and to a minor degree, some DEGs associated with metal transporters and superoxide dismutase activity in root tissues were found. These transcriptomic results suggest that cytoskeleton could be acting as a mechanism of Cu-binding in the root, resulting in a high Cu tolerance response in this metallophyte, which deserve to be analyzed ultra-structurally. Our study contributes to reinforcing the potential of I. cylindrica as a candidate plant species to be used as a phytoremediation agent in Cu-contaminated environments.

Excess Copper-Induced Alterations of Protein Profiles and Related Physiological Parameters in Citrus Leaves

This present study examined excess copper (Cu) effects on seedling growth, leaf Cu concentration, gas exchange, and protein profiles identified by a two-dimensional electrophoresis (2-DE) based mass spectrometry (MS) approach after Citrus sinensis and Citrus grandis seedlings were treated for six months with 0.5 (control), 200, 300, or 400 μM CuCl2. Forty-one and 37 differentially abundant protein (DAP) spots were identified in Cu-treated C. grandis and C. sinensis leaves, respectively, including some novel DAPs that were not reported in leaves and/or roots. Most of these DAPs were identified only in C. grandis or C. sinensis leaves. More DAPs increased in abundances than DAPs decreased in abundances were observed in Cu-treated C. grandis leaves, but the opposite was true in Cu-treated C. sinensis leaves. Over 50% of DAPs were associated with photosynthesis, carbohydrate, and energy metabolism. Cu-toxicity-induced reduction in leaf CO2 assimilation might be caused by decreased abundances of proteins related to photosynthetic electron transport chain (PETC) and CO2 assimilation. Cu-effects on PETC were more pronounced in C. sinensis leaves than in C. grandis leaves. DAPs related to antioxidation and detoxification, protein folding and assembly (viz., chaperones and folding catalysts), and signal transduction might be involved in Citrus Cu-toxicity and Cu-tolerance.

Exogenous of Indole-3-Acetic Acid Application Alleviates Copper Toxicity in Spinach Seedlings by Enhancing Antioxidant Systems and Nitrogen Metabolism

Indole-3-acetic acid (IAA) is a potential mediator in the protection of plants from copper (Cu) toxicity and the enhancement of Cu tolerance. In this paper, spinach (Spinacia oleracea L.) seedlings were cultivated in soil containing 700 mg kg−1 Cu and the leaves of seedlings were sprayed with different concentrations of IAA. Exogenous IAA treatment reduced the malondialdehyde (MDA) concentrations in Cu-stressed seedlings and increased biomass, proline content, and the activities of antioxidant enzymes. Exogenous IAA treatment also increased the levels of nitrogen (N) assimilation compounds and the activities of N-metabolizing enzymes, but reduced NH4+ content. Notably, lower concentrations of IAA (10–40 mg L−1) increased the Cu concentrations in roots and reduced the Cu concentrations in leaves, while higher concentrations of IAA (50 mg L−1) reduced the Cu concentrations in both roots and leaves to the lowest levels. The findings indicated that the application of IAA reduced Cu accumulation, alleviated Cu toxicity, and enhanced Cu tolerance in spinach seedlings. IAA application could be used as an alternative strategy for reducing Cu accumulation in vegetable crops and for remediating Cu-contaminated soil, in turn reducing the hazardous effects of heavy metal contamination on human health and the environment.

Silicon and Iron Differently Alleviate Copper Toxicity in Cucumber Leaves

Copper (Cu) toxicity in plants may lead to iron (Fe), zinc (Zn) and manganese (Mn) deficiencies. Here, we investigated the effect of Si and Fe supply on the concentrations of micronutrients and metal-chelating amino acids nicotianamine (NA) and histidine (His) in leaves of cucumber plants exposed to Cu in excess. Cucumber (Cucumis sativus L.) was treated with 10 µM Cu, and additional 100 µM Fe or/and 1.5 mM Si for five days. High Cu and decreased Zn, Fe and Mn concentrations were found in Cu treatment. Additional Fe supply had a more pronounced effect in decreasing Cu accumulation and improving the molar ratio between micronutrients as compared to the Si supply. However, the simultaneous supply of Fe and Si was the most effective treatment in alleviation of Cu-induced deficiency of Fe, Zn and Mn. Additional Fe supply increased the His but not NA concentration, while Si supply significantly increased both NA and His whereby the NA:Cu and His:Cu molar ratios exceeded the control values indicating that Si recruits Cu-chelation to achieve Cu tolerance. In conclusion, Si-mediated alleviation of Cu toxicity was directed toward Cu tolerance while Fe-alleviative effect was due to a dramatic decrease in Cu accumulation.

Copper accumulation, subcellular partitioning and physiological and molecular responses in relation to different copper tolerance in apple rootstocks

To unravel the physiological and molecular regulation mechanisms underlying the variation in copper (Cu)accumulation, translocation and tolerance among five apple rootstocks, seedlings were exposed to either basal or excess Cu. Excess Cu suppressed plant biomass and root architecture, which was less pronounced in Malus prunifolia Borkh., indicating its relatively higher Cu tolerance. Among the five apple rootstocks, M. prunifolia exhibited the highest Cu concentration and bio-concentration factor in roots but the lowest translocation factor, indicating its greater ability to immobilize Cu and restrict translocation to the aerial parts. Higher Cu concentration in cell wall fraction but lower Cu proportion in membrane-containing and organelle-rich fractions were found in M. prunifolia. Compared with the other four apple rootstocks under excess Cu conditions, M. prunifolia had a lower increment of hydrogen peroxide in roots and leaves and malondialdehyde in roots, but higher concentrations of carbohydrates and enhanced antioxidants. Transcript levels of genes involved in Cu uptake, transport and detoxification revealed species-specific differences that are probably related to alterations in Cu tolerance. M. prunifolia had relatively higher gene transcript levels including copper transporters 2 (COPT2), COPT6 and zinc/iron-regulated transporter-related protein 2 (ZIP2), which probably took part in Cu uptake, and C-type ATP-binding cassette transporter 2 (ABCC2), copper chaperone for Cu/Zn superoxide dismutase (CCS), Cu/Zn superoxide dismutase 1 (CSD1) and metallothionein 2 (MT2) probably implicated in Cu detoxification, and relatively lower mRNA levels of yellow stripe-like transporter 3 (YSL3) and heavy metal ATPase 5 (HMA5) involved in transport of Cu to aerial parts. These results suggest that M. prunifolia is more tolerant to excess Cu than the other four apple rootstocks under the current experimental conditions, which is probably attributed to more Cu retention in roots, subcellular partitioning, well-coordinated antioxidant defense mechanisms and transcriptional expression of genes involved in Cu uptake, translocation and detoxification.

Premature failure of utility poles in Switzerland and Germany related to wood decay basidiomycetes

In contact with soil, copper (Cu) formulations as preservatives are expected to inhibit wood decay by fungi and other soil-borne microorganisms. However, Cu-resistant brown-rot (BR) fungi lead to premature failures of utility poles at some sites. In this study, the service lives of 111 utility poles of Norway spruce (Picea abies (L.) H. Karst) (73 from Switzerland and 38 from Germany) impregnated with Cu-based wood preservatives were investigated. Three segments of each utility pole were analyzed. The severity of decay was dependent on the preservative formulation. BR fungi and in particular Antrodia species were predominantly isolated from utility poles that were not treated with a co-biocide, e.g. boron (B). Cu-sensitivity of several isolated BR fungi was confirmed in studies on Cu-amended medium and in Cu-treated wood. Isolates of Fibroporia vaillantii and Serpula himantioides showed a higher Cu-tolerance than the highly Cu-tolerant Empa isolate Rhodonia placenta (Empa 45) or Antrodia serialis.