Thermal Reversion Research Articles

Divergence in red light responses associated with thermal reversion of phytochrome B between high- and low-latitude species.

Phytochromes play a central role in mediating adaptive responses to light and temperature throughout plant life cycles. Despite evidence for adaptive importance of natural variation in phytochromes, little information is known about molecular mechanisms that modulate physiological responses of phytochromes in nature. We show evolutionary divergence in physiological responses relevant to thermal stability of a physiologically active form of phytochrome (Pfr) between two sister species of Brassicaceae growing at different latitudes. The higher latitude species (Cardamine bellidifolia; Cb) responded more strongly to light-limited conditions compared with its lower latitude sister (C.nipponica; Cn). Moreover, CbPHYB conferred stronger responses to both light-limited and warm conditions in the phyB-deficient mutant of Arabidopsis thaliana than CnPHYB: that is Pfr CbphyB was more stable in nuclei than CnphyB. Our findings suggest that fine tuning Pfr stability is a fundamental mechanism for plants to optimise phytochrome-related traits in their evolution and adapt to spatially varying environments, and open a new avenue to understand molecular mechanisms that fine tune phytochrome responses in nature.

Synthesis of Anthracene‐Based Cyclic π‐Clusters and Elucidation of their Properties Originating from Congested Aromatic Planes

AbstractSynthesis and properties of anthracene‐based cyclic π‐clusters which possess two and four anthracene units are discussed. The optimal cyclization conditions were determined based on a nickel(0)‐mediated reaction that afforded a cyclic anthracene dimer as the major product. Bringing two anthracene planes in close proximity in a face‐to‐face manner resulted in red‐shifted absorption owing to the narrowing of the HOMO–LUMO gap. The cyclic anthracene dimer exhibits multi‐stimuli responsiveness due to high π‐congestion. For example, photoirradiation on the anthracene dimer affords its photoisomer having C−C bonds that are longer than 1.65 Å, which can undergo thermal reversion under gentle heating. This enabled mechanochromism of the photoisomer (colorless) to the original anthracene dimer (red). Photoisomerization was also observed in the crystalline state, accompanied by crystal jumping or collapsing, that is, the photosalient effect.

Synthesis of Anthracene-Based Cyclic π-Clusters and Elucidation of their Properties Originating from Congested Aromatic Planes.

Synthesis and properties of anthracene-based cyclic π-clusters which possess two and four anthracene units are discussed. The optimal cyclization conditions were determined based on a nickel(0)-mediated reaction that afforded a cyclic anthracene dimer as the major product. Bringing two anthracene planes in close proximity in a face-to-face manner resulted in red-shifted absorption owing to the narrowing of the HOMO-LUMO gap. The cyclic anthracene dimer exhibits multi-stimuli responsiveness due to high π-congestion. For example, photoirradiation on the anthracene dimer affords its photoisomer having C-C bonds that are longer than 1.65 Å, which can undergo thermal reversion under gentle heating. This enabled mechanochromism of the photoisomer (colorless) to the original anthracene dimer (red). Photoisomerization was also observed in the crystalline state, accompanied by crystal jumping or collapsing, that is, the photosalient effect.

Effects of mechanical constraint on thermally induced reverse martensitic transformation in granular shape memory ceramic packings

Zirconia-based ceramics exhibit shape memory and superelastic effects based on the reversible martensitic transformation between tetragonal and monoclinic crystal structures. In the form of granular packings, these shape memory ceramics can be scaled up for bulk applications despite their intrinsic brittleness, while displaying drastically different transformation characteristics than the monolithic counterparts. Here, we present a comparative study to understand the thermally induced reverse martensitic transformation in granular packings and the influence of mechanical constraints. This study employs ZrO2–CeO2 shape memory ceramics of the same composition but with different degrees of mechanical constraints. The explored material forms include loose and jammed granular packings, themselves consisting of polycrystalline or single crystal particles, as well as sintered bulk polycrystals. Except for the latter, no endothermic peak is observed in the heat flow measurement of the reverse transformation process. This unusual behavior is shown to stem from the weak inter-particle mechanical constraint and the transformation heterogeneity among individual particles, rather than stress relaxation or particle rearrangement. To compare, conspicuous endothermic peaks only appear in bursting-type transformations under a strong mechanical constraint. For granular packings, the intra-particle mechanical constraint does not affect the presence of any endothermic peaks in thermal reversion but can influence the austenite start temperature.

Measurement of glass-transition temperature of thermoreversible photochromic materials based on mechanochemical amorphization

Photochromic diarylethene (DAE) materials are expected to be applied to various organic devices, and it is important to measure the glass transition temperature (Tg) in order to evaluate their stability. Although DAEs have excellent thermal stability, their colored state has a high melting point, and either a thermal cyclo-reversion reaction or decomposition occurs during the differential scanning calorimetry (DSC) measurement. Tgs in the colored state, therefore, are hard to measure. In this study, we succeeded in measuring Tgs of the colored state with DSC by preparing the colored amorphous state based on a mechanochemical effect for various DAEs. In addition, the relationship between the isomerization ratio and Tg was determined by combining the amorphization and the thermal reversion reaction near the melting point. It was found that Tg changed monotonically according to the isomerization ratio.

Phytochrome B and PCH1 protein dynamics store night temperature information.

Plants experience temperature fluctuations during the course of the daily cycle, and although stem growth responds rapidly to these changes we largely ignore whether there is a short-term memory of previous conditions. Here we show that nighttime temperatures affect the growth of the hypocotyl of Arabidopsis thaliana seedlings not only during the night but also during the subsequent photoperiod. Active phytochrome B (phyB) represses nighttime growth and warm temperatures reduce active phyB via thermal reversion. The function of PHOTOPERIODIC CONTROL OF HYPOCOTYL1 (PCH1) is to stabilise active phyB in nuclear bodies but, surprisingly, warmth reduces PCH1 gene expression and PCH1 stability. When phyB was active at the beginning of the night, warm night temperatures enhanced the levels of nuclear phyB and reduced hypocotyl growth rate during the following day. However, when end-of-day far-red light minimised phyB activity, warm night temperatures reduced the levels of nuclear phyB and enhanced the hypocotyl growth rate during the following day. This complex growth pattern was absent in the phyB mutant. We propose that temperature-induced changes in the levels of PCH1 and in the size of the physiologically relevant nuclear pool of phyB amplify the impact of phyB-mediated temperature sensing.

SYNTHESIS, STRUCTURAL CHARACTERIZATION, cis–trans ISOMERISM, AND DFT STUDIES ON cis-BIS(N,N-DI-METHYL-N′-BENZOYLTHIOUREATO) PALLADIUM(II)

The cis-[Pd(L-κS,O)2] complex is synthesized by the addition of N,N-dimethyl-N′-benzoylthiourea (HL) to $$\text{PdCl}_{4}^{2-}$$ in an acetonitrile:water mixture. The complex is characterized by the elemental analysis and NMR spectroscopy. The single crystal X-ray diffraction analysis shows that the complex crystallizes in the monoclinic P21/c space group, with Pd(II) being bonded in a six-membered chelate. Irradiation of an acetonitrile solution of cis-[Pd(L-κS,O)2] with visible light produces a geometric trans-[Pd(L-κS,O)2] isomer that undergoes a thermal reversion in the absence of light. The relative energies, electronic absorption spectra, and electron density distribution for the cis-[Pd(L-κS,O)2] and trans-[Pd(L-κS,O)2] complexes are calculated using the density functional calculations (DFT) at the B3LYP/CEP31-G level of theory.

PCH1 and PCHL Directly Interact with PIF1, Promote Its Degradation, and Inhibit Its Transcriptional Function during Photomorphogenesis

PHOTOPERIODIC CONTROL OF HYPOCOTYL 1 (PCH1) and PCH1-LIKE (PCHL) were shown to directly bind to phytochrome B (phyB) and suppress phyB thermal reversion, resulting in plants with dramatically enhanced light sensitivity. Here, we show that PCH1 and PCHL also positively regulate various light responses, including seed germination, hypocotyl gravitropism, and chlorophyll biosynthesis, by physically interacting with PHYTOCHROME INTERACTING FACTOR 1 (PIF1) and CONSTITUTIVE PHOTOMORPHOGENIC 1 (COP1). PCH1 and PCHL interact with PIF1 both in the dark and light, and regulate PIF1 abundance. Moreover, PCH1 and PCHL facilitate the physical interaction between phyB and PIF1 in vivo to promote the light-induced degradation of PIF1. PCH1 and PCHL also inhibit the DNA-binding ability of PIF1 to negatively regulate the expressions of PIF1 target genes. In addition, PCH1 and PCHL interact with COP1 and undergo degradation through the 26S proteasome pathway in the dark. Consistently, pch1 suppresses cop1 phenotype in darkness. Collectively, our study reveals a novel mechanism by which PCH1 and PCHL regulate diverse light responses not only by stabilizing phyB Pfr form but also by directly interacting with PIF1 and COP1, providing a molecular understanding of the control of hypocotyl growth by these proteins.

Synthesis, structural characterization, cis—trans isomerism, and DFT studies on cis-bis(N,N-di-methyl-N'-benzoylthioureato)palladium(II)

The cis-[Pd(L-κS,O)2] complex is synthesized by the addition of N,N-dimethyl-N'-benzoyl­thiourea(HL) to PdCl42– in an acetonitrile:water mixture. The complex is characterized by the elemental analysis and NMR spectroscopy. The single crystal X-ray diffraction analysis shows that the complex crystallizes in the monoclinic P21/c space group, with Pd(II) being bonded in a six-membered chelate. Irradiation of an acetonitrile solution of cis-[Pd(L-κS,O)2] with visible light produces a geometric trans-[Pd(L-κS,O)2] isomer that undergoes a thermal reversion in the absence of light. The relative energies, electronic absorption spectra, and electron density distribution for the cis-[Pd(L-κS,O)2] and trans-[Pd(L-κS,O)2] complexes are calculated using the density functional calculations (DFT) at the B3LYP/CEP31-G level of theory.

Synthesis and characterization of bis(4-amino-2-bromo-6-methoxy)azobenzene derivatives

Aminoazobenzene derivatives with four ortho substituents with respect to the N–N double bond are a relatively unexplored class of azo compounds that show promise for use as photoswitches in biology. Tetra-ortho-methoxy-substituted aminoazobenzene compounds in particular can form azonium ions under physiological conditions and exhibit red-light photoswitching. Here, we report the synthesis and characterization of two bis(4-amino-2-bromo-6-methoxy)azobenzene derivatives. These compounds form red-light-absorbing azonium ions, but only under very acidic conditions (pH < 1). While the low pKa makes the azonium form unsuitable, the neutral versions of these compounds undergo trans-to-cis photoisomerization with blue-green light and exhibit slow (τ1/2 ≈ 10 min) thermal reversion and so may find applications under physiological conditions.

Photoreversible interconversion of a phytochrome photosensory module in the crystalline state

A major barrier to defining the structural intermediates that arise during the reversible photointerconversion of phytochromes between their biologically inactive and active states has been the lack of crystals that faithfully undergo this transition within the crystal lattice. Here, we describe a crystalline form of the cyclic GMP phosphodiesterases/adenylyl cyclase/FhlA (GAF) domain from the cyanobacteriochrome PixJ in Thermosynechococcus elongatus assembled with phycocyanobilin that permits reversible photoconversion between the blue light-absorbing Pb and green light-absorbing Pg states, as well as thermal reversion of Pg back to Pb. The X-ray crystallographic structure of Pb matches previous models, including autocatalytic conversion of phycocyanobilin to phycoviolobilin upon binding and its tandem thioether linkage to the GAF domain. Cryocrystallography at 150 K, which compared diffraction data from a single crystal as Pb or after irradiation with blue light, detected photoconversion product(s) based on Fobs - Fobs difference maps that were consistent with rotation of the bonds connecting pyrrole rings C and D. Further spectroscopic analyses showed that phycoviolobilin is susceptible to X-ray radiation damage, especially as Pg, during single-crystal X-ray diffraction analyses, which could complicate fine mapping of the various intermediate states. Fortunately, we found that PixJ crystals are amenable to serial femtosecond crystallography (SFX) analyses using X-ray free-electron lasers (XFELs). As proof of principle, we solved by room temperature SFX the GAF domain structure of Pb to 1.55-Å resolution, which was strongly congruent with synchrotron-based models. Analysis of these crystals by SFX should now enable structural characterization of the early events that drive phytochrome photoconversion.

Differential phosphorylation of the N-terminal extension regulates phytochrome B signaling.

Phytochrome B (phyB) is an excellent light quality and quantity sensor that can detect subtle changes in the light environment. The relative amounts of the biologically active photoreceptor (phyB Pfr) are determined by the light conditions and light independent thermal relaxation of Pfr into the inactive phyB Pr, termed thermal reversion. Little is known about the regulation of thermal reversion and how it affects plants' light sensitivity. In this study we identified several serine/threonine residues on the N-terminal extension (NTE) of Arabidopsis thaliana phyB that are differentially phosphorylated in response to light and temperature, and examined transgenic plants expressing nonphosphorylatable and phosphomimic phyB mutants. The NTE of phyB is essential for thermal stability of the Pfr form, and phosphorylation of S86 particularly enhances the thermal reversion rate of the phyB Pfr-Pr heterodimer in vivo. We demonstrate that S86 phosphorylation is especially critical for phyB signaling compared with phosphorylation of the more N-terminal residues. Interestingly, S86 phosphorylation is reduced in light, paralleled by a progressive Pfr stabilization under prolonged irradiation. By investigating other phytochromes (phyD and phyE) we provide evidence that acceleration of thermal reversion by phosphorylation represents a general mechanism for attenuating phytochrome signaling.

Molecular shape under far‐red light and red light‐induced association of Arabidopsis phytochrome B

Phytochrome B (phyB) is a plant photoreceptor protein that regulates various photomorphogenic responses to optimize plant growth and development. PhyB exists in two photoconvertible forms: a red light-absorbing (Pr) and a far-red light-absorbing (Pfr) form. Therefore, to understand the mechanism of phototransformation, the structural characterization of full-length phyB in these two forms is necessary. Here, we report the molecular structure of Arabidopsisthaliana phyB in Pr form and the molecular properties of the Pfr form determined by small-angle X-ray scattering coupled with size-exclusion chromatography. In solution, the Pr form associated as a dimer with a radius of gyration of 50Å. The molecular shape was a crossed shape, in which the orientation of the photosensory modules differed from that in the crystal structure of dimeric photosensory module. PhyB exhibited structural reversibility in the Pfr-to-Pr phototransformation and thermal reversion from Pfr to Pr in the dark. In addition, Pfr only exhibited nonspecific association, which distinguished molecular properties of Pfr form from those of the inactive Pr form.

Kraft pulp bleaching with a P-stage catalyzed by both bicarbonate and TAED

Peroxide bleaching of softwood and hardwood (eucalypt) kraft pulps was performed in solutions of sodium bicarbonate (NaHCO3), sodium carbonate (Na2CO3), and sodium hydroxide (NaOH). The conventional P stage (hydrogen peroxide + sodium hydroxide; H2O2 + NaOH) was the most effective brightening system without an additional activator. However, peroxide activation by bicarbonate anion (HCO3–) was obvious in all cases where NaHCO3 or Na2CO3 was used. When N,N,N’,N’-tetraacetylethylenediamine (TAED) was added to the bleaching system, Na2CO3 as the alkali source afforded equal or slightly higher bleached brightness compared to NaOH usage for both the softwood and hardwood pulps. This outcome is attributed to simultaneous peroxide activation by HCO3 and TAED. When applied to the eucalypt pulp, the H2O2/Na2CO3/TAED bleaching system also decreased the brightness loss due to thermal reversion.

Role of the Propionic Side Chains for the Photoconversion of Bacterial Phytochromes.

Bacteriophytochromes harboring a biliverdin IXα (BV) chromophore undergo photoinduced reaction cascades to switch between physiologically inactive and active states. Employing vibrational spectroscopic and computational methods, we analyzed the role of propionic substituents of BV in the transformations between parent states Pr and Pfr in prototypical (Agp1) and bathy (Agp2) phytochromes from Agrobacterium fabrum. Both proteins form adducts with BV monoesters (BVM), esterified at propionic side chain B (PsB) or C (PsC), but in each case, only one monoester adduct is reactive. In the reactive Agp2-BVM-B complex (esterified at ring B), the Pfr dark state displays the structural properties characteristic of bathy phytochromes, including a protonated PsC. As in native Agp2, PsC is deprotonated in the final step of the Pfr phototransformation. However, the concomitant α-helix/β-sheet secondary structure change of the tongue is blocked at the stage of unfolding of the coiled loop region. This finding and the shift of the tautomeric equilibrium of BVM toward the enol form are attributed to the drastic changes in the electrostatic potential. The calculations further suggest that deprotonation of PsC and the protonation state of His278 control the reactivity of the enol tautomer, thereby accounting for the extraordinarily slow thermal reversion. Although strong perturbations of the electrostatic potential are also found for Agp1-BVM, the consequences for the Pr-to-Pfr phototransformation are less severe. Specifically, the structural transition of the tongue is not impaired and thermal reversion is even accelerated. The different response of Agp1 and Agp2 to monoesterification of BV points to different photoconversion mechanisms.

Controlled photoisomerization in acrylic copolymer nanoparticles based on spironaphthoxazine for reduced thermal reversion

Stimuli-responsive materials based on spirooxazines are new class of smart materials that have received great deal of attentions in the recent decades. Spirooxazine derivatives, as photochromic compounds, represent many advantages like high photofatigue resistance. But, they usually suffer from very short life time of the colored form and quick reversion to its colorless isomer which limits their application at ambient conditions. Here, spironaphthoxazine (SNO) and spironaphthoxazine acrylate (SNO-Ac, as the monomer) were synthesized and characterized by FTIR and 1H NMR spectroscopy. Then, temperature-dependent photochromic behavior of SNO solution in different solvents was studied by UV–Vis spectroscopy at room and low temperatures. The absorption band relating to the obtained meronaphthoxazine (MNO) in 500–600 nm was not clearly observed after UV irradiation (365 nm) at room temperature, while an intensive absorption was appeared at −30 and −20 °C. SNO-Ac was then copolymerized with methyl methacrylate through solution and miniemulsion polymerizations. The obtained nanoparticles (ranging from 64 to 94 nm in size) represented reduced thermal reversion in comparison with the solution-based copolymers, but still not enough for room-temperature observations. Thereafter, an evident coloration was gained at normal conditions by introducing butyl acrylate to the above-mentioned copolymeric latex. The obtained flexible copolymer chains were found to facilitate SNO to MNO isomerization and the kinetic of photoisomerization in such systems were studied comprehensively. This resulted in photothermal stability enhancement of MNO at ambient conditions by their embedding in the prepared acrylic terpolymer nanoparticle to decrease its decoloration rate as a novel approach in such systems.

PCH1 regulates light, temperature, and circadian signaling as a structural component of phytochrome B-photobodies in Arabidopsis

The members of the phytochrome (phy) family of bilin-containing photoreceptors are major regulators of plant photomorphogenesis through their unique ability to photointerconvert between a biologically inactive red light-absorbing Pr state and an active far-red light-absorbing Pfr state. While the initial steps in Pfr signaling are unclear, an early event for the phyB isoform after photoconversion is its redistribution from the cytoplasm into subnuclear foci known as photobodies (PBs), which dissipate after Pfr reverts back to Pr by far-red irradiation or by temperature-dependent nonphotochemical reversion. Here we present evidence that PHOTOPERIODIC CONTROL OF HYPOCOTYL 1 (PCH1) functions both as an essential structural component of phyB-containing PBs and as a direct regulator of thermal reversion that is sufficient to stabilize phyB as Pfr in vitro. By examining the genetic interaction between a constitutively active phyBY276H-YFP allele (YHB-YFP) and PCH1, we show that the loss of PCH1 prevents YHB from coalescing into PBs without affecting its nuclear localization, whereas overexpression of PCH1 dramatically increases PB levels. Loss of PCH1, presumably by impacting phyB-PB assembly, compromises a number of events elicited in YHB-YFP plants, including their constitutive photomorphogenic phenotype, red light-regulated thermomorphogenesis, and input of phyB into the circadian clock. Conversely, elevated levels of both phyB and PCH1 generate stable, yet far-red light-reversible PBs that persisted for days. Collectively, our data demonstrate that the assembly of PCH1-containing PBs is critical for phyB signaling to multiple outputs and suggest that altering PB dynamics could be exploited to modulate plant responses to light and temperature.

Nucleophilic addition of phosphorus(iii) derivatives to squaraines: colorimetric detection of transition metal-mediated or thermal reversion.

Nucleophilic addition of phosphorus(iii) agents to the electrophilic core of intensely colored squaraine dyes gives a bleached zwitterionic adduct in good to excellent yields (up to 99%) at room temperature. The process can be reversed by adding specific transition metal complexes with high phosphorous(iii) affinity.

Phytochrome B dynamics departs from photoequilibrium in the field.

Vegetation shade is characterized by marked decreases in the red/far-red ratio and photosynthetic irradiance. The activity of phytochrome in the field has typically been described by its photoequilibrium, defined by the photochemical properties of the pigment in combination with the spectral distribution of the light. This approach represents an oversimplification because phytochrome B (phyB) activity depends not only on its photochemical reactions but also on its rates of synthesis, degradation, translocation to the nucleus, and thermal reversion. To account for these complex cellular reactions, we used a model to simulate phyB activity under a range of field conditions. The model provided values of phyB activity that in turn predicted hypocotyl growth in the field with reasonable accuracy. On the basis of these observations, we define two scenarios, one is under shade, in cloudy weather, at the extremes of the photoperiod or in the presence of rapid fluctuations of the light environment caused by wind-induced movements of the foliage, where phyB activity departs from photoequilibrium and becomes affected by irradiance and temperature in addition to the spectral distribution. The other scenario is under full sunlight, where phyB activity responds mainly to the spectral distribution of the light.

The Structure of Cis-2,2′-Azopyridine in the Solid State

Descriptions of azoheteroaromatic compounds in their cis-isomeric form in the solid state are almost non-existent. Cis-2,2′-azopyridine is a known compound which structure differentiates from the trans-isomer based on spectroscopic evidence (UV–Vis absorption, 1H, and 13C NMR spectroscopy) and with studies regarding its stability, thermal reversion to the trans-isomer, and dipole moment in solution. However, the structure of this compound in the solid state has been unresolved. Herein, we present the crystal structure of cis-2,2′-azopyridine that forms red prismatic crystals in the monoclinic space group C2/c. The unit cell parameters are: a = 23.485 (4), b = 5.9241 (12), c = 21.656 (4) A, and V = 2717.1 (9) A3 with 12 molecules per unit cell. Molecules are arranged as two helical conformers (P and M) with C–N=N–C torsion angles and aromatic ring planes’ angles comparable to those reported for cis-azobenzene. The rare structural determination of an azoheteroaromatic cis-isomer of 2-azopyridine that was synthesized, isolated and crystallized is presented. X-ray structure analysis reveals that the molecule forms dimeric complexes containing both M- and P-helical conformers as well as infinite stacked columns in the solid state through a combination of C–H⋯N hydrogen bonds, C–H⋯π interactions and lone pair⋯π contacts.