A New Player in Jasmonate-Mediated Stomatal Closure: The Arabidopsis thaliana Copper Amine Oxidase β

Plant defence responses to adverse environmental conditions include different stress signalling, allowing plant acclimation and survival. Among these responses one of the most common, immediate, and effective is the modulation of the stomatal aperture, which integrates different transduction pathways involving hydrogen peroxide (H2O2), calcium (Ca2+), nitric oxide (NO), phytohormones and other signalling components. The Arabidopsis thaliana copper amine oxidases β (AtCuAOβ) encodes an apoplastic CuAO expressed in guard cells and root protoxylem tissues which oxidizes polyamines to aminoaldehydes with the production of H2O2 and ammonia. Here, its role in stomatal closure, signalled by the wound-associated phytohormone methyl-jasmonate (MeJA) was explored by pharmacological and genetic approaches. Obtained data show that AtCuAOβ tissue-specific expression is induced by MeJA, especially in stomata guard cells. Interestingly, two Atcuaoβ T-DNA insertional mutants are unresponsive to this hormone, showing a compromised MeJA-mediated stomatal closure compared to the wild-type (WT) plants. Coherently, Atcuaoβ mutants also show compromised H2O2-production in guard cells upon MeJA treatment. Furthermore, the H2O2 scavenger N,N1-dimethylthiourea (DMTU) and the CuAO-specific inhibitor 2-bromoethylamine (2-BrEtA) both reversed the MeJA-induced stomatal closure and the H2O2 production in WT plants. Our data suggest that AtCuAOβ is involved in the H2O2 production implicated in MeJA-induced stomatal closure.

Microcrystal preparation for serial femtosecond X-ray crystallography of bacterial copper amine oxidase

Recent advances in serial femtosecond X-ray crystallography (SFX) using X-ray free-electron lasers have paved the way for determining radiation-damage-free protein structures under nonfreezing conditions. However, the large-scale preparation of high-quality microcrystals of uniform size is a prerequisite for SFX, and this has been a barrier to its widespread application. Here, a convenient method for preparing high-quality microcrystals of a bacterial quinoprotein enzyme, copper amine oxidase from Arthrobacter globiformis, is reported. The method consists of the mechanical crushing of large crystals (5–15 mm3), seeding the crushed crystals into the enzyme solution and standing for 1 h at an ambient temperature of ∼26°C, leading to the rapid formation of microcrystals with a uniform size of 3–5 µm. The microcrystals diffracted X-rays to a resolution beyond 2.0 Å in SFX measurements at the SPring-8 Angstrom Compact Free Electron Laser facility. The damage-free structure determined at 2.2 Å resolution was essentially identical to that determined previously by cryogenic crystallography using synchrotron X-ray radiation.

Plant Copper Amine Oxidases: Key Players in Hormone Signaling Leading to Stress-Induced Phenotypic Plasticity

Polyamines are ubiquitous, low-molecular-weight aliphatic compounds, present in living organisms and essential for cell growth and differentiation. Copper amine oxidases (CuAOs) oxidize polyamines to aminoaldehydes releasing ammonium and hydrogen peroxide, which participates in the complex network of reactive oxygen species acting as signaling molecules involved in responses to biotic and abiotic stresses. CuAOs have been identified and characterized in different plant species, but the most extensive study on a CuAO gene family has been carried out in Arabidopsis thaliana. Growing attention has been devoted in the last years to the investigation of the CuAO expression pattern during development and in response to an array of stress and stress-related hormones, events in which recent studies have highlighted CuAOs to play a key role by modulation of a multilevel phenotypic plasticity expression. In this review, the attention will be focused on the involvement of different AtCuAOs in the IAA/JA/ABA signal transduction pathways which mediate stress-induced phenotypic plasticity events.

Resistance to Growth of Molds for Wood Modified with Hydrophobic Hybrid Silica Gel Containing Copper Amine Complexes

Complexation copper with amine provides an effective strategy for fixation copper in wood, while hydrophobic modification improves the dimensional stability of wood. Thus, a combination of complexation and hydrophobization is expected to enhance the efficiency of copper-based biocides. In this study, hydrophobic hybrid silica gel containing copper amine complexes (MACu) was prepared through an in situ sol-gel process in wood using methyltriethoxysilane (MTES), 3-amino-propyltriethoxysilane (APTES), and copper chloride. The resistance to growth of molds for MACu modified wood (Populus tomentosa) was measured according to ASTM D3273-16. A leaching resistance test was carried out in accordance with AWPA E11-16. The results showed that only Aspergillus niger covered the surface of untreated wood blocks and no mold grew on the MACu surface even after the leaching test. MACu xerogel and MACu wood were further characterized by SEM-EDS, FTIR, and XPS. A possible schematic diagram of the reaction mechanism was proposed to explain the high-efficiency anti-mold performance of MACu wood.

Proton conductive polyoxoniobate frameworks constructed from nanoscale {Nb68O200} cages

Nanoscale {Nb68O200} cages have been successfully employed as flexible and stable secondary building units to combine with bridging copper–amine complexes to construct two proton conductive polyoxoniobate frameworks, demonstrating a promising strategy for making new porous materials.

The effects of pH on copper leaching from wood treated with copper amine-based preservatives

As pH of leaching medium is an important factor in the leaching of wood preservative components, its effects on leaching should be quantified to ensure environmentally safe use of treated wood. In this study, the effects of pH on leaching of copper from wood treated with copper amine-based preservatives [alkaline copper quat (ACQ)-2, bis-(N-cyclohexyldiazeniumdioxy)-copper (CuHDO)-3, and copper azole (CUAZ)-3] were evaluated in comparison with wood treated with chromated copper arsenate (CCA)-3. Radiata pine sapwood blocks treated with these preservatives were leached at five pH levels (3.0, 3.5, 4.0, 4.5, and 6.5). The leached blocks were subjected to laboratory-scale decay tests using two brown-rot fungi. The blocks treated with copper amine-based preservatives leached significant amounts of copper at pH levels below 4.0. At all pH levels, the CuHDO-3-treated samples generally leached the most copper, followed by the samples treated with ACQ-2, CUAZ-3, and CCA-3. When the treated blocks were leached at pH 3.0, the degradation of hemicelluloses, which can chemically adsorb copper, was confirmed through Fourier transform infrared attenuated total reflectance (FTIR-ATR) analysis. Moreover, X-ray photoelectron spectroscopy (XPS) analysis indicated that the ratio of precipitates of the remaining copper in the treated wood severely decreased after leaching at pH levels below 4.0. Subsequent reduction in the biological effectiveness of wood treated with copper amine-based preservatives was not hardly observed after leaching at pH levels 4.0 or above. These results indicate that copper loss at pH levels 4.0 or above is not great enough to cause public concern about environmental problems and reduction of biological efficacy in practical applications.

Decarboxylative Propargylation/Hydroamination/Aromatization Enabled by Copper/ Amine Cooperative Catalysis: Construction of Cyclopenta[b]indole Derivatives

An efficient decarboxylative cycloaddition of 2,3-dioxopyrrolidines and ethynyl benzoxazinanones has been established by cooperative copper/amine catalysis. A copper– allenylidene complex and enolate intermediate, each catalytically generated from distinct substrates, underwent a cascade propargylation/hydroamination/aromatization process to construct a big library of cyclopenta[b]indole derivatives with good to excellent yields and excellent diastereoselectivity.