Biotransformation of copper oxide nanoparticles by the pathogenic fungus Botrytis cinerea

The effects of copper (Cu) on microorganisms have been studied for decades due to its strong antimicrobial activity. Nowadays, emerging technologies are developing new antimicrobial compounds such as CuO and Cu nanoparticles (NPs), or products with their inclusions. In this study two plant pathogenic fungi, Alternaria alternata and Botrytis cinerea, were exposed to Cu in either ionic (Cu2+) or microparticle (MP, CuO) or nanoparticle (NP, Cu or CuO) form, in solid and liquid culturing media. B. cinerea proved to be resistant to CuO and Cu NPs and CuO MPs in comparison to A. alternata as shown by pronounced growth and lower levels of lipid peroxidation. B. cinerea grown in the presence of CuO and Cu NPs and CuO MPs on solid medium formed a blue compound at the fungal/ culturing medium interface, followed by a Cu depletion zone. The blue compound was characterized as Cu-oxalate by Cu-K EXAFS. In B. cinerea, pronounced activity of catechol-type siderophores and/or organic acid secretion apparently induces leaching and mobilization of Cu ions from the CuO MPs, CuO and Cu NPs and their further complexation with extracellularly secreted oxalic acid. As such, the pathogenic fungus B. cinerea may be used for copper extraction and/or purification and synthesis of different materials

Linking microbiome and hyperaccumulation in plants

Hyperaccumulating plants can take up extraordinarily large concentrations of one or more metal(loid)s from the soil and accumulate it/them in the aboveground tissues without exibiting any visible toxicity symptoms. Among more than 700 plant taxa reported to have evolved this unique phenotype, the most common is the hyperaccumulation of nickel (Ni), and less common is the hyperaccumulation of arsenic (As), cadmium (Cd), cobalt (Co), chromium (Cr), copper (Cu), manganese (Mn), lead (Pb), antimony (Sb), selenium (Se), thallium (Tl) or zinc (Zn). Metal(loid) hyperaccumulation is a result of several independent evolutionary events and despite considerable efforts, none of the proposed hypotheses on the environmental constraints driving these events has been supported fully to date. Among several tolerance strategies enabling hyperaccumulation is the allocation of metal(loid)s to competent cell types, typically away from photosynthetic apparatus, to limit damage to plant metabolism. Recently, the involvement of microorganisms colonizing roots in hyperacumulation phenomenon has achieved increased attention due to their role in the mobilization of metal(loid)s in the soil. The complex interactions between hyperaccumulation and belowground microbiome are of primary interest for phytoremediation, a promising green technology for removing or immobilisation of metal(loid)s in the soil with the help of plants. In this review, we discuss and complement current reports on the contribution of microorganisms to metal(loid) hyperaccumulation

Arbuscular mycorrhizal fungi (AMF) root colonization dynamics of Molinia caerulea (L.) Moench. in grasslands and post-industrial sites

This is an accepted manuscript of an article published by Elsevier in Ecological Engineering on 05/08/2016, available online: https://doi-org.ezproxy.wlv.ac.uk/10.1016/j.ecoleng.2016.06.029 The accepted version of the publication may differ from the final published version.The aims of this studies were: (i) to examine the influence of heavy metal content (Zn, Cd, Pb, Fe, Cu) and other physico-chemical soil parameters on the level of root colonization of Molinia caerulea and (ii) to relate root colonisation parameters and soil variables to Molinia caerulea abundance in two contrasting habitats (grasslands and heavy metal contaminated sites). The sites differ significantly in terms of bio-available heavy metal contents, particularly Zn (34 times more than grasslands), soil texture, CaCO3, organic matter (LOI%), Mg and nitrate content. Principal Component Analysis showed the strong negative correlations between frequency of mycorrhization (F), arbuscular abundance (A%) and intensity of root cortex colonisation (M%) and concentration of bio-available Zn and Cd. Moreover, no positive correlation between root colonization of Molinia and its abundance was found. The frequency of mycorrhization of root fragments (F%) was only slightly different between these two habitats, whereas the intensity of root cortex colonisation (M%) and relative arbuscular abundance (A%) were significantly lower (3 and 4 times respectively) on the post-industrial sites. The bioavailable Zn content in the substratum of post-industrial sites was strongly negatively correlated with species richness, Shannon diversity index and Evenness. In contrast, these relationships were not statistically significant in grasslands. Based on obtained results we could draw a model of possible relationships between root colonization of Molinia, HM content and Molinia abundance on grasslands and post-industrial sites. Bioavailable Zn content in the soil is a one of main factors influencing the Molinia community diversity. In the grasslands, lower amounts of bioavailable Zn, resulted in higher species richness (R) and species diversity (H) which in turn lead to higher root colonization. On the other hand, on the post-industrial sites, the elevated bioavailable Zn content strongly decreases the plant species richness (R) and species diversity (H) and this caused the decline in root colonization parameters. The low species richness on Zn-polluted sites allowed Molinia to reach higher abundance since the competition with other species is reduced

Germination characteristics of Salicornia patula Duval-Jouve, S. emerici Duval-Jouve, and S. veneta Pign. et Lausi and their occurrence in Croatia

According to recent molecular analyses of Salicornia, we revised the annual glassworts from the Croatian coast, classified until now only as Salicornia europaea. Two species, a diploid Salicornia patula and a tetraploid S. emerici were recognized. They can be easily distinguished by floral characters, but not only by their habitus, which varies extremely according to environmental factors. Both species differ also in seed morphology. Salicornia patula has dimorphic seeds, with larger central seeds reaching high germination rates. Germination patterns helped to explain the habitat preferences. The species rarely co-occur, however. The rare S. patula occupies drier habitats, on coastal mudflats or sands that are irregularly inundated. It occurs within the assoc. Suaedo maritimae-Salicornietum patulae. Salicornia emerici occupies the lowest coastal mudflats, regularly inundated, where nutrient-rich conditions prevail, and forms an almost monotypical assoc. Salicornietum emerici. Due to the synonymy of S. veneta with S. emerici, we exclude the occurrence of S. veneta in Croatia as an independent taxon

Environmental metabarcoding reveals contrasting belowground and aboveground fungal communities from poplar at a Hg phytomanagement site

Characterization of microbial communities in stressful conditions at a field level is rather scarce, especially when considering fungal communities from aboveground habitats. We aimed at characterizing fungal communities from different poplar habitats at a Hg-contaminated phytomanagement site by using Illumina-based sequencing, network analysis approach, and direct isolation of Hg-resistant fungal strains. The highest diversity estimated by the Shannon index was found for soil communities, which was negatively affected by soil Hg concentration. Among the significant correlations between soil operational taxonomic units (OTUs) in the co-occurrence network, 80% were negatively correlated revealing dominance of a pattern of mutual exclusion. The fungal communities associated with Populus roots mostly consisted of OTUs from the symbiotic guild, such as members of the Thelephoraceae, thus explaining the lowest diversity found for root communities. Additionally, root communities showed the highest network connectivity index, while rarely detected OTUs from the Glomeromycetes may have a central role in the root network. Unexpectedly high richness and diversity were found for aboveground habitats, compared to the root habitat. The aboveground habitats were dominated by yeasts from the Lalaria, Davidiella, and Bensingtonia genera, not detected in belowground habitats. Leaf and stem habitats were characterized by few dominant OTUs such as those from the Dothideomycete class producing mutual exclusion with other OTUs. Aureobasidium pullulans, one of the dominating OTUs, was further isolated from the leaf habitat, in addition to Nakazawaea populi species, which were found to be Hg resistant. Altogether, these findings will provide an improved point of reference for microbial research on inoculation-based programs of tailings dumps

New insights into globoids of protein storage vacuoles in wheat aleurone using synchrotron soft X-ray microscopy

Mature developed seeds are physiologically and biochemically committed to store nutrients, principally as starch, protein, oils, and minerals. The composition and distribution of elements inside the aleurone cell layer reflect their biogenesis, structural characteristics, and physiological functions. It is therefore of primary importance to understand the mechanisms underlying metal ion accumulation, distribution, storage, and bioavailability in aleurone subcellular organelles for seed fortification purposes. Synchrotron radiation soft X-ray full-field imaging mode (FFIM) and low-energy X-ray fluorescence (LEXRF) spectromicroscopy were applied to characterize major structural features and the subcellular distribution of physiologically important elements (Zn, Fe, Na, Mg, Al, Si, and P). These direct imaging methods reveal the accumulation patterns between the apoplast and symplast, and highlight the importance of globoids with phytic acid mineral salts and walls as preferential storage structures. C, N, and O chemical topographies are directly linked to the structural backbone of plant substructures. Zn, Fe, Na, Mg, Al, and P were linked to globoid structures within protein storage vacuoles with variable levels of co-localization. Si distribution was atypical, being contained in the aleurone apoplast and symplast, supporting a physiological role for Si in addition to its structural function. These results reveal that the immobilization of metals within the observed endomembrane structures presents a structural and functional barrier and affects bioavailability. The combination of high spatial and chemical X-ray microscopy techniques highlights how in situ analysis can yield new insights into the complexity of the wheat aleurone layer, whose precise biochemical composition, morphology, and structural characteristics are still not unequivocally resolved

Identification of conserved secondary structures and expansion segments in enod40 RNAs reveals new enod40 homologues in plants

enod40 is a plant gene that participates in the regulation of symbiotic interaction between leguminous plants and bacteria or fungi. Furthermore, it has been suggested to play a general role in non-symbiotic plant development. Although enod40 seems to have multiple functions, being present in many land plants, the molecular mechanisms of its activity are unclear; they may be determined though, by short peptides and/or RNA structures encoded in the enod40 genes. We utilized conserved RNA structures in enod40 sequences to search nucleotide sequence databases and identified a number of new enod40 homologues in plant species that belong to known, but also, to yet unknown enod40-containing plant families. RNA secondary structure predictions and comparative sequence analysis of enod40 RNAs allowed us to determine the most conserved structural features, present in all known enod40 genes. Remarkably, the topology and evolution of one of the conserved structural domains are similar to those of the expansion segments found in structural RNAs such as rRNAs, RNase P and SRP RNAs. Surprisingly, the enod40 RNA structural elements are much more stronger conserved than the encoded peptides. This finding suggests that some general functions of enod40 gene could be determined by the encoded RNA structure, whereas short peptides may be responsible for more diverse functions found only in certain plant families

Fungal root endophyte associations of plants endemic to the Pamir Alay Mountains of Central Asia

The fungal root endophyte associations of 16 species from 12 families of plants endemic to the Pamir Alay Mountains of Central Asia are presented. The plants and soil samples were collected in Zeravshan and Hissar ranges within the central Pamir Alay mountain system. Colonization by arbuscular mycorrhizal fungi (AMF) was found in 15 plant species; in 8 species it was of the Arum type and in 4 of the Paris type, while 3 taxa revealed intermediate arbuscular mycorrhiza (AM) morphology. AMF colonization was found to be absent only in Matthiola integrifolia, the representative of the Brassicaceae family. The AM status and morphology are reported for the first time for all the species analyzed and for the genera Asyneuma, Clementsia, and Eremostachys. Mycelia of dark septate endophytes (DSE) accompanied the AMF colonization in ten plant species. The frequency of DSE occurrence in the roots was low in all the plants, with the exception of Spiraea baldschuanica. However, in the case of both low and higher occurrence, the percentage of DSE root colonization was low. Moreover, the sporangia of Olpidium spp. were sporadically found inside the root epidermal cells of three plant species. Seven AMF species (Glomeromycota) found in the trap cultures established with soils surrounding roots of the plants being studied were reported for the first time from this region of Asia. Our results provide information that might well be of use to the conservation and restoration programmes of these valuable plant species. The potential application of beneficial root-inhabiting fungi in active plant protection projects of rare, endemic and endangered plants is discussed