Structurally Defined Water-Soluble Metallofullerene Derivatives towards Biomedical Applications.

Endohedral metallofullerenes (EMFs) are excellent carriers of rare-earth element (REE) ions in biomedical applications because they preclude the release of toxic metal ions. However, existing approaches to synthesize water-soluble EMF derivatives yield mixtures that inhibit precise drug design. Here we report the synthesis of metallobuckytrio (MBT), a three-buckyball system, as a modular platform to develop structurally defined water-soluble EMF derivatives with ligands by choice. Demonstrated with PEG ligands, the resulting water-soluble MBTs show superb biocompatibility. The Gd MBTs exhibit superior T1 relaxivity than typical Gd complexes, potentially superseding current clinical MRI contrast agents in both safety and efficiency. The Lu MBTs generated reactive oxygen species upon light irradiation, showing promise as photosensitizers. With their modular nature to incorporate other ligands, we anticipate the MBT platform to open new paths towards bio-specific REE drugs.

A novel naturally Phe206Tyr mutation confers tolerance to ALS-inhibiting herbicides in Alopecurus myosuroides.

Herbicide-resistant weeds pose a serious threat to world food production. The rapid and widespread development of target-site based resistance limits the application of herbicides. Alopecurus myosuroides Huds. (blackgrass) has spread rapidly in winter wheat regions in China, and the field recommended dose of ALS herbicides no longer controls blackgrass populations in recent years. A highly resistant population TW18(R) was collected in 2018 from Shandong Province. Dose-response assays showed that the TW18 was resistant to mesosulfuron-methyl, flucarbazone-sodium, and imazethapyr, with resistance index values of 5.96, 6.1, and 4.09, respectively. DNA sequencing of the TW18 population revealed a Phe206Tyr (F206Y) mutation in the ALS, which was not yet reported. Blackgrass ALS gene with the F206Y mutation (R gene) was expressed in Arabidopsis and rice. Transgenic studies have shown that both Arabidopsis and rice expressing this R gene have resistance to imazethapyr. However, it did not confer resistance to tribenuron methyl and florasulam in transgenic Arabidopsis. This study showed that the F206Y substitution caused herbicide resistance in blackgrass. To our knowledge, this is the first-reported F206Y mutation of a weed species in the natural environment. Transgenic plants showed this functional site could be utilized to generate imazethapyr-resistant rice to control herbicide-resistant weed damage.

Transcriptome Profiling of Cu Stressed Petunia Petals Reveals Candidate Genes Involved in Fe and Cu Crosstalk.

Copper (Cu) is an essential element for most living plants, but it is toxic for plants when present in excess. To better understand the response mechanism under excess Cu in plants, especially in flowers, transcriptome sequencing on petunia buds and opened flowers under excess Cu was performed. Interestingly, the transcript level of FIT-independent Fe deficiency response genes was significantly affected in Cu stressed petals, probably regulated by basic-helix-loop-helix 121 (bHLH121), while no difference was found in Fe content. Notably, the expression level of bHLH121 was significantly down-regulated in petals under excess Cu. In addition, the expression level of genes related to photosystem II (PSII), photosystem I (PSI), cytochrome b6/f complex, the light-harvesting chlorophyll II complex and electron carriers showed disordered expression profiles in petals under excess Cu, thus photosynthesis parameters, including the maximum PSII efficiency (FV/FM), nonphotochemical quenching (NPQ), quantum yield of the PSII (ΦPS(II)) and photochemical quenching coefficient (qP), were reduced in Cu stressed petals. Moreover, the chlorophyll a content was significantly reduced, while the chlorophyll b content was not affected, probably caused by the increased expression of chlorophyllide a oxygenase (CAO). Together, we provide new insight into excess Cu response and the Cu-Fe crosstalk in flowers.

Unexpected Formation of Metallofulleroids from Multicomponent Reactions, with Crystallographic and Computational Studies of the Cluster Motion.

New multicomponent reactions involving an isocyanide, terminal or internal alkynes, and endohedral metallofullerene (EMF) Lu3 N@C80 yield metallofulleroids which are characterized by mass-spectrometry, HPLC, and multiple 1D and 2D NMR techniques. Single crystal studies revealed one ketenimine metallofulleroid has ordered Lu3 N cluster which is unusual for EMF monoadducts. Computational analysis, based on crystallographic data, confirm that the endohedral cluster motion is controlled by the position of the exohedral organic appendants. Our findings provide a new functionalization reaction for EMFs, and a potential facile approach to freeze the endohedral cluster motion at relatively high temperatures.

Controlled Synthesis of Nitrogen-Doped Graphene on Ruthenium from Azafullerene.

The controlled synthesis of high-quality nitrogen (N) doped single layer graphene on the Ru(0001) surface has been achieved using the N-containing sole precursor azafullerence (C59NH). The synthesis process and doping properties have been investigated on the atomic scale by combining scanning tunneling microscopy and X-ray photoelectron spectroscopy measurements. We find for the first time that the concentration of N-related defects on the N-doped graphene/Ru(0001) surface is tunable by adjusting the dosage of sole precursor and the number of growth cycles. Two primary types of N-related defects have been observed. The predominant bonding configuration of N atoms in the obtained graphene layer is pyridinic N. Our findings indicate that the synthesis from heteroatom-containing sole precursors is a very promising approach for the preparation of doped graphene materials with controlled doping properties.

Oxygen-Delivery Materials: Synthesis of an Open-Cage Fullerene Derivative Suitable for Encapsulation of H2 O2 and O2.

An open-cage [60]fullerene was prepared through a multiple-step sequence based on peroxide-mediated cage-opening reactions. Key steps include repeated C60 -sensitized singlet-oxygen oxidation of electron-rich amino enol double bonds to form two lactone and two lactam moieties on the rim of the orifice. Single-crystal X-ray analysis shows that the 22-membered orifice has an ellipsoid shape with the major axis at 6.7 Šand the minor axis at 3.5 Å. Encapsulation of H2 O2 was observed under atmospheric pressure at room temperature. Oxygen is also effectively trapped during the process.

Two Silene vulgaris copper transporters residing in different cellular compartments confer copper hypertolerance by distinct mechanisms when expressed in Arabidopsis thaliana.

Silene vulgaris is a metallophyte of calamine, cupriferous and serpentine soils all over Europe. Its metallicolous populations are hypertolerant to zinc (Zn), cadmium (Cd), copper (Cu) or nickel (Ni), compared with conspecific nonmetallicolous populations. These hypertolerances are metal-specific, but the underlying mechanisms are poorly understood. We investigated the role of HMA5 copper transporters in Cu-hypertolerance of a S. vulgaris copper mine population. Cu-hypertolerance in Silene is correlated and genetically linked with enhanced expression of two HMA5 paralogs, SvHMA5I and SvHMA5II, each of which increases Cu tolerance when expressed in Arabidopsis thaliana. Most Spermatophytes, except Brassicaceae, possess homologs of SvHMA5I and SvHMA5II, which originate from an ancient duplication predating the appearance of spermatophytes. SvHMA5II and the A. thaliana homolog AtHMA5 localize in the endoplasmic reticulum and upon Cu exposure move to the plasma membrane, from where they are internalized and degraded in the vacuole. This resembles trafficking of mammalian homologs and is apparently an extremely ancient mechanism. SvHMA5I, instead, neofunctionalized and always resides on the tonoplast, likely sequestering Cu in the vacuole. Adaption of Silene to a Cu-polluted soil is at least in part due to upregulation of two distinct HMA5 transporters, which contribute to Cu hypertolerance by distinct mechanisms.

[60]Fullerene-Based Macrocycle Ligands.

Macrocycle ligands have three or more donor sites. Selective replacement of skeleton carbon atoms by heteroatoms and vacancies in C60 could lead to various macrocycle ligands with a cage-shaped backbone. Theoretical calculations indicate that such C60 -based macrocycle ligands are as stable as C60 thermodynamically according to their similar HOMO-LUMO gaps. The synthesis of these ligands is a challenging task. Nevertheless important progresses have been reported. This concept article focuses on the structures of possible C60 -based macrocycle ligands and related synthetic results.

Synthesis of C70 -Based Fluorophores through Sequential Functionalization to Form Isomerically Pure Multiadducts.

Selective addition to the C70 cage divides its π-conjugated system into various smaller π-conjugated systems with enhanced fluorescent properties. Key reactions include chlorination, methoxylation, ozonation, and Bingel or Bingel-Hirsch reactions. The maximum emission wavelength of the C70 multiadducts ranges from 450 to 655 nm. Among the C70 multiadducts, C70 (OMe)8 (C(COOEt)2 )3 showed the highest quantum yield (ΦF =0.18) and the largest Δ[λmax (emission)- λmax (absorption)] (402 nm), with maximum emission at 655 nm.

Evolution of tonoplast P-ATPase transporters involved in vacuolar acidification.

Petunia mutants (Petunia hybrida) with blue flowers defined a novel vacuolar proton pump consisting of two interacting P-ATPases, PH1 and PH5, that hyper-acidify the vacuoles of petal cells. PH5 is similar to plasma membrane H(+) P3A -ATPase, whereas PH1 is the only known eukaryoticP3B -ATPase. As there were no indications that this tonoplast pump is widespread in plants, we investigated the distribution and evolution of PH1 and PH5. We combined database mining and phylogenetic and synteny analyses of PH1- and PH5-like proteins from all kingdoms with functional analyses (mutant complementation and intracellular localization) of homologs from diverse angiosperms. We identified functional PH1 and PH5 homologs in divergent angiosperms. PH5 homologs evolved from plasma membrane P3A -ATPases, acquiring an N-terminal tonoplast-sorting sequence and new cellular function before angiosperms appeared. PH1 is widespread among seed plants and related proteins are found in some groups of bacteria and fungi and in one moss, but is absent in most algae, suggesting that its evolution involved several cases of gene loss and possibly horizontal transfer events. The distribution of PH1 and PH5 in the plant kingdom suggests that vacuolar acidification by P-ATPases appeared in gymnosperms before flowers. This implies that, next to flower color determination, vacuolar hyper-acidification is required for yet unknown processes.

Selective Multiamination of C70 Leading to Curved π†Systems with 60, 58, 56, and 50 π†Electrons.

Secondary aliphatic amines add to a pole pentagon of [70]fullerene in the presence of N-fluorobenzenesulfonimide to form cyclopentadienyl-type adducts, C70(NSO2 Ph)(NR(1)R(2))4 (1), which can be converted into analogous C70 derivatives such as C70 (NHSO2 Ph)(NHTol)5 (2). Further addition reactions of either 1 or 2 take place selectively at the opposite pole pentagon of the C70 cage, thus forming curved π systems with a reduced number of π electrons, and the products include a dodecakis-adduct with a Vögtle belt motif.

Fullerene-Based Macro-Heterocycle Prepared through Selective Incorporation of Three†N and Two O†Atoms into C60.

A 14-membered heterocycle is created on the C60 cage skeleton through a multistep procedure. Key steps involve repeated PCl5 -induced hydroxylamino N-O bond cleavage leading to insertion of nitrogen atoms, and also piperidine-induced peroxo O-O bond cleavage leading to insertion of oxygen atoms. The hetero atoms form one pyrrole, two pyran, and one diazepine rings in conjunction with the C60 skeleton carbon atoms. The fullerene-based macrocycle showed unique reactivities towards fluoride ion and copper salts.

Selective addition of secondary amines to C60: formation of penta- and hexaamino[60]fullerenes.

Secondary amines are well-known to add to [60]fullerene to form the tetraamino epoxy adduct C60(O)(NR1R2)4 under both photolysis and thermal conditions in the presence of oxygen. We have now found that pentaamino hydroxyl adduct C60(OH)(NR1R2)5 and hexaamino adduct C60(NR1R2)6 can be formed as the major products in the dark in the presence of oxygen. Key steps of the reaction mechanism probably involve repeated oxygen oxidation of the radical ion pair between fullerene and amines.

Waste-green infrastructure nexus: Green roof promotion by digestate and digestate biochar from food waste.

Waste-Green Infrastructure Nexus is crucial to mitigate carbon emissions in waste disposal and promote eco-functions of green infrastructure in a circular bio-economy. Our purpose is to verify the feasibility of the nexus via "food waste anaerobic digestion - digestate/digestate biochar - green roof promotion". The results found that food waste digestate and digestate biochar significantly promoted green roof plant growth, evapotranspiration, rainwater retention, runoff reduction, and prevention of nutrient leaching. Digestate treatments were better than digestate biochar for the green roof promotion. The promotion ranked consistently with 20 % digestate > 10 % digestate > 20 % digestate biochar > 10 % digestate biochar > control in stolon growth, leaf emergence, branching of Paspalum vaginatum, green roof establishment, rainwater retention, runoff reduction, and the leaching of nitrogen, phosphorus, potassium. This study demonstrated that food waste could be regenerated to promote urban green infrastructure to form a circular bio-economy by the Waste-Green Infrastructure Nexus.

Bio-pump cadmium phytoextraction efficiency promoted by phytohormones in Festuca arundinacea.

The leaves of Festuca arundinacea can excrete cadmium (Cd) out onto the leaf surface, leading to a bio-pump phytoremediation strategy based on "root uptake-root-to-leaf translocation-leaf excretion". However, the bio-bump efficiency of soil Cd is a limiting factor for the implementation of this novel technology. Bio-bump remediation involves the bioprocess of plant root uptake from soil, root-to-leaf translocation, and leaf hydathode excretion. Here we show the significant effects of phytohormones in regulating the bio-pump phytoextraction efficiency. The results showed that salicylic acid and ethylene enhanced the whole process of Cd root uptake, root-to-leaf translocation, and leaf excretion, promoting the bio-pump phytoextraction efficiency by 63.6% and 73.8%, respectively. Gibberellin also greatly promoted Cd translocation, leaf excretion, and phytoextraction, but did not significantly impact Cd root uptake. Our results indicate that salicylic acid and ethylene could be recommended to promote bio-pump phytoextraction efficiency in F. arundinacea. Gibberellin might be used for a short-term promotion of the leaf Cd excretion.

A WRKY gene from Tamarix hispida, ThWRKY4, mediates abiotic stress responses by modulating reactive oxygen species and expression of stress-responsive genes.

WRKY transcription factors are involved in various biological processes, such as development, metabolism and responses to stress. However, their exact roles in abiotic stress tolerance are largely unknown. Here, we demonstrated a working model for the function of a WRKY gene (ThWRKY4) from Tamarix hispida in the stress response. ThWRKY4 is highly induced by abscisic acid (ABA), salt and drought in the early period of stress (stress for 3, 6, or 9 h), which can be regulated by ABF (ABRE binding factors) and Dof (DNA binding with one finger), and also can be crossregulated by other WRKYs and autoregulated as well. Overexpression of ThWRKY4 conferred tolerance to salt, oxidative and ABA treatment in transgenic plants. ThWRKY4 can improve the tolerance to salt and ABA treatment by improving activities of superoxide dismutase and peroxidase, decreasing levels of O2 (-) and H2O2, reducing electrolyte leakage, keeping the loss of chlorophyll, and protecting cells from death. Microarray analyses showed that overexpression of ThWRKY4 in Arabidopsis leads to 165 and 100 genes significantly up- and downregulated, respectively. Promoter scanning analysis revealed that ThWRKY4 regulates the gene expression via binding to W-box motifs present in their promoter regions. This study shows that ThWRKY4 functions as a transcription factor to positively modulate abiotic stress tolerances, and is involved in modulating reactive oxygen species.

Controllable Mono-/Dialkenylation of Benzyl Thioethers through Rh-Catalyzed Aryl C-H Activation.

Under solvent control: Benzyl thioethers were alkenylated in excellent yields with broad substrate scope and the selectivity (mono- vs. disubstituted product) was controlled by the solvent and ratio of reactants. Sequential alkenylation with two different alkenes was also carried out in a one-pot process. In addition, the thioether directing group was removed in a one-pot process with simultaneous hydrogenation of the double bond to give the toluene derivatives.

A versatile Agrobacterium-mediated transient gene expression system for herbaceous plants and trees.

Plant transient expression is a powerful method used widely for the functional characterization of genes and protein production. In comparison with stable transformation, it has the advantages of being simple, quick, economical, and effective. In the present study, we developed a novel transient gene expression system based on Agrobacterium-mediated transformation. This system is simple and convenient and allows for high transient expression levels. Hyperosmotic pretreatment of plants significantly improved the transient expression in this system. Furthermore, other factors, including acetosyringone concentration, cocultivation time, and Agrobacterium cell density, significantly influenced transient expression efficiency. The results showed that this method is suitable for use with herbaceous plants (such as tobacco and Arabidopsis) and trees (such as birch, poplar, tamarisk, cork, willow, and aralia), suggesting that it may be applied widely in plant transient expression studies.

Synthesis of [60]- and [70]Fullerene-Fused Tetrahydroquinoxaline Derivatives by Oxidative [4 + 2] Cycloaddition with Unusual Reactivity and Regioselectivity.

The oxidative [4 + 2] reaction of o-phenylenediamine-derived disulfonamides with fullerene C60 and C70 is reported, in which electron-deficient reactants showed high reactivity. The reaction of C70 exhibited unusual regioselectivity, yielding a [5,6]-adduct as the major product, which was characterized by 1H, 13C NMR and single-crystal X-ray diffraction. DFT calculations revealed the reaction is an inverse-electron-demand Diels-Alder (IEDDA) reaction, and the [5,6]-adduct of C70 is a kinetic product.

Cadmium binding during leaf senescence in Festuca arundinacea: Promotion phytoextraction efficiency by harvesting dead leaves.

Phytoextraction by harvesting dead leaves is a novel cadmium (Cd) phytoremediation strategy in tall fescue (Festuca arundinacea), which provides feasibility for the phytoremediation of Cd-polluted soils and cleaner food production. The highest Cd in dead leaves is the result of Cd accumulation during the process of leaf senescence. However, it is not known the mechanism of Cd accumulation during the leaf senescence, which limits the phytoextraction efficiency of this technology. In this study, we found that the contents of phytochelatins (PC), glutathione (GSH), and non-protein thiols (NPT) were increased during the process of leaf senescence and Cd stress significantly promoted PC, GSH, and NPT. Transcriptome analysis showed that the pathway of glutathione metabolism was significantly enriched in the senescent leaf under Cd stress. 19 genes encoding GST, enzymes catalyzing GSH-Cd binding, were up-regulated in the senescent leaf. The increases of PC, GSH, and NPT in the senescent leaf for Cd-binding could be from the pathways of the protein degradation rather than their synthesis, because genes encoding cysteine protease (catalyzes protein degradation) were significantly promoted, but both GSH synthetase (GS) and PC synthetase (PCS) did not show the significant changes between the young and senescent leaves. Our results indicated that Cd accumulation during the leaf senescence could be the result of the promotion of Cd-binding by PC, GSH, and NPT, which provide insights into the regulatory mechanism and further genetic engineering to promote the phytoextraction efficiency by harvesting dead leaves in tall fescue.