pH‐dependent Interaction of Rhodopsin with Cyanidin‐3‐glucoside. 2. Functional Aspects†

Anthocyanins are a class of phytochemicals that confer color to flowers, fruits, vegetables and leaves. They are part of our regular diet and serve as dietary supplements because of numerous health benefits, including improved vision. Recent studies have shown that the anthocyanin cyanidin‐3‐O‐glucoside (C3G) increased regeneration of the dim‐light photoreceptor rhodopsin (Matsumoto et al. [2003] J. Agric. Food Chem., 51 , 3560–3563). In an accompanying study (Yanamala et al. [2009] Photochem. Photobiol.), we show that C3G directly binds to rhodopsin in a pH‐dependent manner. In this study, we investigated the functional consequences of C3G binding to rhodopsin. As observed previously in rod outer segments, regeneration of purified rhodopsin in detergent micelles is also accelerated in the presence of C3G. Thermal denaturation and stability studies using circular dichroism, fluorescence and UV/visible absorbance spectroscopy show that C3G exerts a destabilizing effect on rhodopsin structure while it only modestly alters G‐protein activation and the rates at which the light‐activated Metarhodopsin II state decays to opsin and free retinal. These results indicate that the mechanism of C3G‐enhanced regeneration may be based on changes in opsin structure promoting access to the retinal binding pocket.     

Possible Involvement of a Tetrahydrobiopterin in Photoreception for UV‐B‐induced Anthocyanin Synthesis in Carrot

Our previous studies of action spectra for UV‐B‐induced anthocyanin accumulation in cultured carrot cells indicated that a reduced form of pterin, possibly tetrahydrobiopterin, contributes to UV‐B photoreception. In this report, we provide additional evidence for the involvement of pterin in UV‐B light sensing. UV‐B‐induced phenylalanine ammonia‐lyase (PAL) activity was considerably suppressed by N‐acetylserotonin (an inhibitor of tetrahydrobiopterin biosynthesis), and this suppression was partially recovered by adding biopterin or tetrahydrobiobiopterin. In addition, protein(s) specifically bound to biopterin were detected by radiolabeling experiments in N‐acetylserotonin‐treated cells. Furthermore, diphenyleneiodonium, a potent inhibitor of electron transfer, completely suppressed UV‐B‐induced PAL activity. These results suggest the occurrence of an unidentified UV‐B photoreceptor (other than UVR8, the tryptophan‐based UV‐B sensor originally identified in Arabidopsis) with reduced pterin in carrot cells. After reexamining published action spectra, we suggest that anthocyanin synthesis is coordinately regulated by these two UV‐B sensors.     

Reflectance and Cyclic Electron Flow as an Indicator of Drought Stress in Cotton (Gossypium hirsutum)

The response and the functioning of the photosynthetic machinery of cotton, Gossypium hirsutum during water stress was studied by leaf optical properties, linear (ETRII) and cyclic electron flow (CEF) and chlorophyll a fluorescence. We observed that in G. hirsutum, during water limitation, Chlorophyll b showed the best correlation with reflectance at 731 nm and is a better indicator of drought. F_v/F_m was observed to be very insensitive to mild water stress. However, during severe water stress the leaves exhibit considerable inhibition in F_v/F_m and an increase in anthocyanin levels by about 20‐fold. CEF was very responsive to mild water stress. The mild drought stress caused large decrease in the ability of the leaves to utilize the light energy. Photosystem I and photosystem II is protected from photoinhibition by high CEF and nonphotochemical quenching under mild water stress. While during severe drought stress, linear electron flow showed a sharp decrease in comparison to CEF. CEF play a major role in G. hirsutum leaves during mild as well as under severe water stress condition and is thus a good indicator of water stress.     

EXTENSION GROWTH AND ANTHOCYANIN RESPONSES OF PHOTOMORPHOGENIC TOMATO MUTANTS TO CHANGES IN THE PHYTOCHROME: PHOTOEQUILIBRIUM DURING THE DAILY PHOTOPERIOD

Four genotypes of tomato (Lycopersicon esculentum Mill.) in the genetic background ‘Aha Craig’ were used: an aurea (au) mutant, deficient in the bulk light-labile phytochrome pool; a high pigment (hp) mutant, showing exaggerated phytochrome responses at the time of de-etiolation; the au,hp double mutant and the isogenic wild type (WT). A dramatic increase in plant height resulting from an increase in the length of all internodes for each of the genotypes studied was observed upon reduction of the red light: far-red light photon ratio (R:FR) from 6.90 to 0.13 by addition of FR for the whole photoperiod. A concomitant increase in leaf length was also observed. Since au and au,hp mutants, deficient in the bulk light-labile phytochrome pool, respond to this reduction in the R:FR these data demonstrate that the phytochrome pool that mediates this response is present and fully functional. Anthocyanin was detectable in the comparably developed young growing leaves of the WT and hp mutant under the high R:FR, but not in the au and au,hp mutants, suggesting that the potential for anthocyanin synthesis is correlated with the presence of the bulk light-labile phytochrome pool. The kinetics of anthocyanin decrease in the young growing leaves were investigated in the hp mutant and the results suggest a very rapid cessation of flavonoid biosynthesis upon reduction of the R:FR. The functions of different phytochrome types are discussed.     

Effects of Suplementary Ultraviolet‐B Irradiance on Maize Yield and Qualities: A Field Experiment¶

Stratospheric ozone depletion has caused an increase in the amount of ultraviolet‐B (UV‐B) radiation reaching the earth's surface. Numerous investigations have demonstrated that the effect of UV‐B enhancements on plants includes reduction in grain yield, alteration in species competition, susceptibility to disease and changes in plant structure and pigmentation. Many experiments examining UV‐B radiation effects on plants have been conducted in growth chambers or greenhouses. It has been questioned whether the effect of UV‐B radiation on plants can be extrapolated to field responses from indoor studies because of the unnaturally high ratios of UV‐B/ ultraviolet‐A radiation (320–400 nm) and UV‐B/photosynthetically active radiation (PAR) in many indoor studies. Field studies on UV‐B radiation effect on plants have been recommended to use the UV and PAR irradiance provided by natural light. This study reports the growth and yield responses of a maize crop exposed to enhanced UV‐B radiation and the UV‐B effects on aize seed qualities under field conditions. Enhanced UV‐B radiation caused a significant reduction in the dry matter accumulation and the maize yield in turn was affected. With increased UV‐B radiation the flavonoid accumulation in maize leaves increased and the contents of chlorophyll a, b and (a+b) of maize leaves were reduced. The levels of protein, sugar and starch of maize seed decreased with enhanced UV‐B radiation, whereas the level of lysine increased with enhanced UV‐B radiation.     

Photocatalytic Self‐Doped SnO2−x Nanocrystals Drive Visible‐Light‐Responsive Color Switching

Visible‐light‐responsive reversible color‐switching systems are attractive to many applications because visible light has superior penetration and causes far less damage to organic molecules than UV. Herein, we report that self‐doping of SnO_2−x nanocrystals with Sn²⁺ red‐shifts their absorption to the visible region and simultaneously produces oxygen vacancies, which can effectively scavenge photogenerated holes and thus enable the color switching of redox dyes using visible light. Wavelength‐selective switching can also be achieved by coupling the photocatalytic activity of the SnO_2−x NCs with the color‐switching kinetics of different redox dyes. The fast light response enables the further fabrication of a solid film that can be repeatedly written on using a visible laser pen or projection printing through a photomask. This discovery represents a big step forward towards practical applications, especially in areas in which safety issues and photodamage by UV light are of concern.     

High red,green, and blue color purity electroluminescence from MEH‐PPV and polyalkyfluorenes‐based bright white polymer light emitting displays

A series of white polymer light emitting displays (PLEDs) based on a polymer blend of polyalkylfluorenes and poly(2‐methoxy‐5,2′‐ethyl‐hexyloxy‐1,4‐phenylene vinylene) (MEH‐PPV) was developed. MEH‐PPV or red light emitting alkyfluorene copolymer (PFR) was blended with blue light emitting alkyfluorene copolymer (PFB), and MEH‐PPV was blended with both green light emitting alkyfluorene copolymer (PFG) and PFB to generate white light emission PLEDs. Low turn on voltage (2.7 V), high brightness (12,149 nits), high efficiency (4.0 cd/A, 4.0 lm/W), and good color purity (Commission Internationale de L'Eclairage (CIE_x,y) co‐ordinates (0.32, 0.34)) were obtained for the white PLEDs based on the PFB and MEH‐PPV polymer blend. Exciplex formation in the interface between PFR and PFB induced a new green emission peak for these two components based white PLEDs. As a result, strong white emission (4078 nits) was obtained by mixing the red, green, and blue (RGB) three primary colors. High color purity of blue (CIE, x = 0.14, y = 0.08), green (CIE, x = 0.32, y = 0.64) and red (CIE, x = 0.67, y = 0.33) emissions was achieved for white PLEDs combining with dielectric interference color‐filters. © 2006 Wiley Periodicals, Inc. J Polym Sci Part B: Polym Phys 45: 330–341, 2007     

The Effects of Ambient Solar UV Radiation on Alkaloid Production by Erythroxylum novogranatense var. novogranatense†

Truxillines are alkaloids produced by Erythroxylum species and are thought to be derived from the UV‐driven dimerization of cinnamoylcocaines. This study was conducted to determine the effects of ambient UV radiation on the production of truxillines in Erythroxylum novogranatense var. novogranatense. Field plants were grown under shelters covered with plastic filters that were transparent to UV radiation, filtered UV‐B, or both filtered UV‐B and UV‐A radiation. The treatments had no significant effect on plant biomass or specific leaf weight. Absorption values in the UV‐C and UV‐A region of acidified‐methanol leaf extracts were higher for plants exposed to UV radiation compared to the no UV radiation treatment. There was a trend in decreasing levels of trans‐cinnamoylcocaine and a statistically significant decrease in levels of cis‐cinnamoylcocaine in the leaves of plants exposed to UV radiation compared to the no UV radiation treatment. Truxilline levels increased in leaves from plants exposed to UV radiation compared to the no UV radiation treatment. Most significantly, the ratio of truxillines to total cinnamoylcocaines in the leaves was affected by UV, increasing with increased UV exposure. The results support the hypothesis that UV radiation is involved in the formation of truxillines from cinnamoylcocaines.     

NMR-based characterization of a novel yellow chlorophyll catabolite, Ed-YCC,isolated from Egeria densa

A novel yellow chlorophyll catabolite, Ed-YCC, was isolated from leaves detached from Egeria densa shoots, in which chlorophyll degradation and anthocyanin synthesis were induced in 0.1 M fructose solution under light illumination as a plant senescence process, a model of autumnal leaf coloration. Structure elucidation was accomplished by various NMR techniques including 2D-INADEQUATE.     

Large‐scale isolation of high‐purity anthocyanin monomers from mulberry fruits by combined chromatographic techniques

Anthocyanins have attracted attention over the past several decades because of their beneficial health effects. In this research, a strategy combining column chromatography and high‐speed countercurrent chromatography was developed for the separation of high‐purity anthocyanin monomers from mulberry fruits. After purification using Amberlite XAD‐7HP column with 80% ethanol (0.1% HCl), a fraction of anthocyanins mixtures with a purity of 68.6% was obtained. High‐speed countercurrent chromatography with a biphasic solvent system of n‐butanol/methyl tert‐butyl ether/acetonitrile/water/trifluoroacetic acid (30:10:10:50:0.05, v/v) was used to separate the anthocyanin monomers. Three monomers of delphinidin‐3‐O‐ rutinoside, cyanidin‐3‐O‐ rutinoside, and cyanidin‐3‐O‐ glucoside were obtained, and identified by ¹H and ¹³C NMR spectroscopy and high‐performance liquid chromatography with electrospray ionization‐mass spectrometry. The method developed in this work can be used to conduct large‐scale separations of anthocyanin monomers from mulberry fruits and other plants.     

Purification and antioxidant capacity analysis of anthocyanin glucoside cinnamic ester isomers from Solanum nigrum fruits

In a recent study, anthocyanins, which have a strong free radical‐scavenging activity, were examined for their potential to effectively prevent cancer. However, clinical trials are limited by the purity of the anthocyanin. Multiple methods are used to extract and purify anthocyanins. Based on previous work on Solanum nigrum, which is a widely distributed plant, in this study, DM130 macroporous resin, Sephadex LH20, and a C18 column were used to separate cis–trans anthocyanin isomers. These anthocyanins constitute the majority of total S. nigrum anthocyanins. The results showed that this “DM130‐LH20‐C18 system” can be used to obtain a cinnamic acid‐derived cis–trans anthocyanin, petunidin‐3‐(p‐coumaroyl)‐rutinoside‐5‐glucoside, with a purity of 98.5%, for effective quantitation. In order to determine the antioxidant ability of the petunidin‐3‐(p‐coumaroyl)‐rutinoside‐5‐glucoside cis–trans isomers, three ordinary methods were adopted. The maximum antioxidant ability of the cis–trans anthocyanin was dozens of times higher than that of vitamin C.     

Cloning of the Orange Light‐Producing Luciferase from Photinus scintillans—A New Proposal on how Bioluminescence Color is Determined

Unlike the enchanting yellow‐green flashes of light produced on warm summer evenings by Photinus pyralis, the most common firefly species in North America, the orange lights of Photinus scintillans are infrequently observed. These Photinus species, and likely all bioluminescent beetles, use the same substrates beetle luciferin, ATP and oxygen to produce light. It is the structure of the particular luciferase enzyme that is the key to determining the color of the emitted light. We report here the molecular cloning of the P. scintillans luc gene and the expression and characterization of the corresponding novel recombinant luciferase enzyme. A comparison of the amino acid sequence with that of the highly similar P. pyralis enzyme and subsequent mutagenesis studies revealed that the single conservative amino acid change tyrosine to phenylalanine at position 255 accounted for the entire emission color difference. Additional mutagenesis and crystallographic studies were performed on a H‐bond network, which includes the position 255 residue and five other stringently conserved beetle luciferase residues, that is proximal to the substrate/emitter binding site. The results are interpreted in the context of a speculative proposal that this network is key to the understanding of bioluminescence color determination.     

Phytochrome Control of Anthocyanin Biosynthesis in Tomato Seedlings: Analysis Using Photomorphogenic Mutants

Anthocyanin biosynthesis has been studied in hypocotyls and whole seedlings of tomato (Lycoperskon esculentum Mill.) wild types (WTs) and photomorphogenic mutants. In white light (WL)/dark (D) cycles the fri¹ mutant, deficient in phytochrome A (phyA), shows an enhancement of anthocyanin accumulation, whereas the tri¹ mutant, deficient in phytochrome Bl (phyBl) has a WT level of anthocyanin. Under pulses of red light (R) or R followed by far-red light (FR) given every 4 h, phyA is responsible for the non-R/FR reversible response, whereas phyBl is partially responsible for the R/FR reversible response. From R and blue light (B) pretreatment studies, B is most effective in increasing phytochrome responsiveness, whereas under R itself it appears to be dependent on the presence of phyBl. Anthocyanin biosynthesis during a 24 h period of monochromatic irradiation at different flu-ence rates of 4 day-old D-grown seedlings has been studied. At 660 nm the fluence rate-response relationships for induction of anthocyanin in the WT are similar, yet complex, showing a low fluence rate response (LFRR) and a fluence rate-dependent high irradiance response (HIR). The high-pigment-1 (hp-1) mutant exhibits a strong amplification of both the LFRR and HIR. The fri¹ mutant lacks the LFRR while retaining a normal HIR. In contrast, a transgenic tomato line overexpressing the oat PHYA3 gene shows a dramatic amplification of the LFRR. The tri¹ mutant, retains the LFRR but lacks the HIR, whereas the fri¹, tri¹ double mutant lacks both components. Only an LFRR is seen at 729 nm in WT; however, an appreciable HIR is observed at 704 nm, which is retained in the tri¹ mutant and is absent in the fri¹ mutant, indicating the labile phyA pool regulates this response component.     

Rapid quantitative analysis of individual anthocyanin content based on high‐performance liquid chromatography with diode array detection with the pH differential method

A new quantitative technique for the simultaneous quantification of the individual anthocyanins based on the pH differential method and high‐performance liquid chromatography with diode array detection is proposed in this paper. The six individual anthocyanins (cyanidin 3‐glucoside, cyanidin 3‐rutinoside, petunidin 3‐glucoside, petunidin 3‐rutinoside, and malvidin 3‐rutinoside) from mulberry (Morus rubra) and Liriope platyphylla were used for demonstration and validation. The elution of anthocyanins was performed using a C₁₈ column with stepwise gradient elution and individual anthocyanins were identified by high‐performance liquid chromatography with tandem mass spectrometry. Based on the pH differential method, the high‐performance liquid chromatography peak areas of maximum and reference absorption wavelengths of anthocyanin extracts were conducted to quantify individual anthocyanins. The calibration curves for these anthocyanins were linear within the range of 10–5500 mg/L. The correlation coefficients (r²) all exceeded 0.9972, and the limits of detection were in the range of 1–4 mg/L at a signal‐to‐noise ratio ≥5 for these anthocyanins. The proposed quantitative analysis was reproducible with good accuracy of all individual anthocyanins ranging from 96.3 to 104.2% and relative recoveries were in the range 98.4–103.2%. The proposed technique is performed without anthocyanin standards and is a simple, rapid, accurate, and economical method to determine individual anthocyanin contents.     

Synthesis of halide anion‐doped bismuth terephthalate hybrids for organic pollutant removal

Halide anion‐doped bismuth terephthalate hybrids were synthesized using a facile solvothermal method. Four series of hybrids doped with halide anions X^? (F^?, Cl^?, Br^? and I^?) were produced by varying the molar ratios (n) of X^? relative to Bi(NO₃)₃ (n = 0.25, 0.5, 0.75 and 1) in dimethylformamide solution. The results indicated that 0.25 equiv. of different halide anion‐doped bismuth terephthalate hybrids, especially BiBDC‐Cl(0.25) and BiBDC‐Br(0.25), exhibited excellent photocatalytic activity under visible light and UV light irradiation. They also exhibited excellent adsorption performance for Rhodamine B which could be attributed to high surface areas and negative charge on the surface of the catalysts. Moreover, the degradation of Rhodamine B under visible light irradiation is a photosensitization process and ^?O₂^? is the most important active species. The halide anion‐doped bismuth terephthalate hybrids are promising photocatalysts for removal of organic pollutants. Copyright © 2016 John Wiley & Sons, Ltd.     

PHYSIOLOGICAL CHARACTERIZATION OF A HIGH-PIGMENT MUTANT OF TOMATO

Abstract— A high-pigment (hp) mutant, which shows exaggerated phytochrome responses and three other genotypes of Lycopersicon esculenrum Mill. cv. Ailsa Craig: the aurea (au) mutant deficient in the bulk light-labile phytochrome (PI) pool, the au, hp double mutant, and their isogenic wild type, were used in this study. Measurements of phytochrome destruction in red light (R) revealed that the exaggerated responses of the hp mutant are not caused by a higher absolute phytochrome level or a reduced rate of phytochrome destruction. Fluence-response relationships for anthocyanin synthesis after a blue-light pretreatment were studied to test if the hp mutant conveys hypersensitivity to the far-red light (FR)-absorbing form of phytochrome (Pfr), i.e. the threshold of Pfr required to initiate the response is lower. The response range for the hp mutant and wild type was identical, although the former exhibited a 6-fold larger response. Moreover, the kinetics of anthocyanin accumulation in continuous R were similar in the wild-type and hp-mutant seedlings, despite the latter accumulating 9-fold more anthocyanin. Since the properties of phytochrome are the same, the hp mutation appears to affect the state of responsiveness amplification, i.e. the same amount of Pfr leads to a higher response in the hp mutant. We therefore propose that the hp mutation is associated with an amplification step in the phytochrome transduction chain. Escape experiments showed that the anthocyanin synthesis after different light pretreatments terminated with a R pulse was still 50% FR reversible after 4–6 h darkness, indicating that the Pfr pool regulating this response must be relatively stable. However, fluence-rate response relationships for anthocyanin synthesis and hypocotyl growth induced by a 24-h irradiation with 451, 539, 649, 693, 704 and 729 nm light showed no or a severely reduced response in the au and au, hp mutants, suggesting the importance of PI in these responses. We therefore propose that the capacity for anthocyanin synthesis (state of responsiveness amplification) could be established by PI, while the anthocyanin synthesis is actually photoregulated via a stable Pfr pool. The Hp gene product is proposed to be an inhibitor of the state of responsiveness amplification for responses controlled by this relatively stable Pfr species.     

Increased Antioxidant Activity and Changes in Phenolic Profile of Kalanchoe pinnata (Lamarck) Persoon (Crassulaceae) Specimens Grown Under Supplemental Blue Light

Antioxidant compounds protect plants against oxidative stress caused by environmental conditions. Different light qualities, such as UV‐A radiation and blue light, have shown positive effects on the production of phenols in plants. Kalanchoe pinnata (Lamarck) Persoon (Crassulaceae) is used for treating wounds and inflammations. Some of these beneficial effects are attributed to the antioxidant activity of plant components. We investigated the effects of blue light and UV‐A radiation supplementation on the total phenol content, antioxidant activity and chromatographic profile of aqueous extracts from leaves of K. pinnata. Monoclonal plants were grown under white light, white plus blue light and white plus UV‐A radiation. Supplemental blue light improved the antioxidant activity and changed the phenolic profile of the extracts. Analysis by HPLC of supplemental blue‐light plant extracts revealed a higher proportion of the major flavonoid quercetin 3‐O‐α‐l ‐arabinopyranosyl (1→2) α‐l ‐rhamnopyranoside, as well as the presence of a wide variety of other phenolic substances. These findings may explain the higher antioxidant activity observed for this extract. Blue light is proposed as a supplemental light source in the cultivation of K. pinnata, to improve its antioxidant activity.     

Improving mechanical properties and chemical resistance of ink‐jet printer color filter by using diblock polymeric dispersants

The diblock copolymers of polystyrene and poly(tert‐butyl acrylate) (PSt‐b‐PtBA) with various molecular weights and hydrophobic/hydrophilic (styrene/acrylic acid) chain length were prepared by atom transfer radical polymerization (ATRP). Selective hydrolysis of the diblock copolymers (PSt‐b‐PtBA) resulted in amphiphilic block copolymers of polystyrene and poly(acrylic acid) (PSt‐b‐PAA). The amphiphilic block copolymers of PSt‐b‐PAA with average molecular weight (M_n) <7500 were proved to be critical in dispersing the pigments of UV curable ink‐jet inks for manufacturing the color filter. Incorporating DB2 diblock copolymer dispersants with styrene/acrylic acid ratio at 1.5 allowed more UV curable compositions in the red and blue inks without deteriorating pigment dispersing stability and jetting properties of the ink‐jet inks. The ink drops can be precisely ejected into the tiny color area. Better properties of the cured red stripe such as nanoindentation hardness and chemical resistance were found. The competing absorption of UV light by the blue pigment hindered the through cure of monomers near the interface between glass substrate and the blue stripe. This leads to lower hardness and poor chemical resistance of the UV cured blue stripe. © 2005 Wiley Periodicals, Inc. J Polym Sci Part B: Polym Phys 43: 3337–3353, 2005