A simple method to visualize the mechanism why Alnus glutinosa remains green during autumn colouration of Sorbus aucuparia

The autumn colouration of two deciduous trees (Sorbus aucuparia and Alnus glutinosa) from northern Finland (65°N) was studied during leaf senescence in autumn 2003. Their leaves were harvested from the end of August until leaf abscission in Sorbus trees at the end of September. The leaves were extracted in ethanol and measured with a spectrophotometer at 663 nm (chlorophyll a), 644 nm (chlorophyll b), 536 (anthocyanins) and 470 nm (carotenoids). The ratios A663/644, A663/470 and A663/536 were calculated to demonstrate the degradation of chlorophyll a in relation to the other studied pigments. The most important results of the present study were as follows: (1) Rapid (within 4 days) visually observed change in Sorbus colour from green to red. (2) Simultaneously occurred significant change in the calculated absorbance ratios in the same species. (3) The distinct difference between the tree species in both variables: Alnus remained green and maintained its absorbance ratios above the threshold ratios of Sorbus throughout the experiment. Sorbus exhibited the reddish colour in a ratio of 0.4 for A663/470 and 4 for A663/536 around the middle of September. The lowest ratios for Alnus remained 0.5 and 6 for A663/470 and for A663/536 (respectively), i.e. the ratios in Alnus never reached the level in Sorbus. Alnus maintained its green colour throughout the studied period, but the Sorbus trees started to turn red in mid-September. Alnus remained green for two reasons: It maintained higher chlorophyll concentrations, and higher ratios of chlorophyll to other pigments.     

Thallus morphology and optical characteristics affect growth and DNA damage by UV radiation in juvenile Arctic Laminaria sporophytes

Growth of young sporophytes of the brown algae Laminaria digitata, L. saccharina and L. solidungula from Spitsbergen were measured in the laboratory after being exposed for 21 days to either photosynthetically active radiation (PAR=P) or to full light spectrum (PAR + UV-A + UV-B=PAB) using of cutoff glass filters. The plants were grown at 8±2°C and 16 h light : 8 h dark cycles with 6 h additional ultraviolet radiation (UVR) exposure in the middle of the light period. Growth was measured every 10 min using growth chambers with online video measuring technique. Tissue morphology and absorption spectra were measured in untreated young sporophytes while chlorophyll (Chl) a content and DNA damage were measured in treated thalli at the end of the experiment. In all species, growth rates were significantly higher in sporophytes exposed to P alone compared to sporophytes exposed to PAB. Tissue DNA damage is dependent on thallus thickness and absorption spectra characteristics of pigments and UV-absorbing compounds. In sporophytes exposed to UVR, energy demands for repair of DNA damage and synthesis of UV-absorbing compounds for protection effectively diverts photosynthate at the expense of growth. Photosynthetic pigment was not significantly different between treatments suggesting a capacity for acclimation to moderate UVR fluence. The general growth pattern in sporophytes exposed to P alone showed an increasing growth rate from the onset of light (0500–0900 hours) to a peak at the middle of the light phase (0900–1500 hours), a decline towards the end of the light phase (1500–2100 hours) and a minimum “low” growth in the dark (2100–0500 hours) relative to growth during the entire light phase. Under PAB, different growth patterns were observed such as growth compensation at night in L. digitata, delayed growth recovery in L. saccharina and minimal but continuous growth in L. solidungula. Growth as an integrative parameter of all physiological processes showed that the effect of UVR is correlated to the depth distribution of these species.     

Isolation and characterization of homodimeric type-I reaction center complex from Candidatus Chloracidobacterium thermophilum, an aerobic chlorophototroph

The recently discovered thermophilic acidobacterium Candidatus Chloracidobacterium thermophilum is the first aerobic chlorophototroph that has a type-I, homodimeric reaction center (RC). This organism and its type-I RCs were initially detected by the occurrence of pscA gene sequences, which encode the core subunit of the RC complex, in metagenomic sequence data derived from hot spring microbial mats. Here, we report the isolation and initial biochemical characterization of the type-I RC from Ca. C. thermophilum. After removal of chlorosomes, crude membranes were solubilized with 0.1% (w/v) n-dodecyl β-D-maltoside, and the RC complex was purified by ion-exchange chromatography. The RC complex comprised only two polypeptides: the reaction center core protein PscA and a 22-kDa carotenoid-binding protein denoted CbpC. The absorption spectrum showed a large, broad absorbance band centered at ～483 nm from carotenoids as well as smaller Q(y) absorption bands at 672 and 812 nm from chlorophyll a and bacteriochlorophyll a, respectively. The light-induced difference spectra of whole cells, membranes, and the isolated RC showed maximal bleaching at 840 nm, which is attributed to the special pair and which we denote as P840. Making it unique among homodimeric type-I RCs, the isolated RC was photoactive in the presence of oxygen. Analyses by optical spectroscopy, chromatography, and mass spectrometry revealed that the RC complex contained 10.3 bacteriochlorophyll a(P), 6.4 chlorophyll a(PD), and 1.6 Zn-bacteriochlorophyll a(P)' molecules per P840 (12.8:8.0:2.0). The possible functions of the Zn-bacteriochlorophyll a(P)' molecules and the carotenoid-binding protein are discussed.     

Functional Comparison of Rod and Cone Gαt on the Regulation of Light Sensitivity

The signaling cascades mediated by G protein-coupled receptors (GPCRs) exhibit a wide spectrum of spatial and temporal response properties to fulfill diverse physiological demands. However, the mechanisms that shape the signaling response of the GPCR are not well understood. In this study, we replaced cone transducin α (cTα) for rod transducin α (rTα) in rod photoreceptors of transgenic mice, which also express S opsin, to evaluate the role of Gα subtype on signal amplification from different GPCRs in the same cell; such analysis may explain functional differences between retinal rod and cone photoreceptors. We showed that ectopically expressed cTα 1) forms a heterotrimeric complex with rod Gβ₁γ₁, 2) substitutes equally for rTα in generating photoresponses initiated by either rhodopsin or S-cone opsin, and 3) exhibited similar light-activated translocation as endogenous rTα in rods and endogenous cTα in cones. Thus, rTα and cTα appear functionally interchangeable. Interestingly, light sensitivity appeared to correlate with the concentration of cTα when expression is reduced below 35% of normal. However, quantification of endogenous cTα concentration in cones showed a higher level to rTα in rods. Thus, reduced sensitivity in cones cannot be explained by reduced coupling efficiency between the GPCR and G protein or a lower concentration of G protein in cones versus rods.     

Photoprotection in plants involves a change in lutein 1 binding domain in the major light-harvesting complex of photosystem II

Nonphotochemical quenching (NPQ) is the fundamental process by which plants exposed to high light intensities dissipate the potentially harmful excess energy as heat. Recently, it has been shown that efficient energy dissipation can be induced in the major light-harvesting complexes of photosystem II (LHCII) in the absence of protein-protein interactions. Spectroscopic measurements on these samples (LHCII gels) in the quenched state revealed specific alterations in the absorption and circular dichroism bands assigned to neoxanthin and lutein 1 molecules. In this work, we investigate the changes in conformation of the pigments involved in NPQ using resonance Raman spectroscopy. By selective excitation we show that, as well as the twisting of neoxanthin that has been reported previously, the lutein 1 pigment also undergoes a significant change in conformation when LHCII switches to the energy dissipative state. Selective two-photon excitation of carotenoid (Car) dark states (Car S(1)) performed on LHCII gels shows that the extent of electronic interactions between Car S(1) and chlorophyll states correlates linearly with chlorophyll fluorescence quenching, as observed previously for isolated LHCII (aggregated versus trimeric) and whole plants (with versus without NPQ).     

Origin of absorption changes associated with photoprotective energy dissipation in the absence of zeaxanthin

To prevent photo-oxidative damage to the photosynthetic membrane in strong light, plants dissipate excess absorbed light energy as heat in a mechanism known as non-photochemical quenching (NPQ). NPQ is triggered by the trans-membrane proton gradient (ΔpH), which causes the protonation of the photosystem II light-harvesting antenna (LHCII) and the PsbS protein, as well as the de-epoxidation of the xanthophyll violaxanthin to zeaxanthin. The combination of these factors brings about formation of dissipative pigment interactions that quench the excess energy. The formation of NPQ is associated with certain absorption changes that have been suggested to reflect a conformational change in LHCII brought about by its protonation. The light-minus-dark recovery absorption difference spectrum is characterized by a series of positive and negative bands, the best known of which is ΔA(535). Light-minus-dark recovery resonance Raman difference spectra performed at the wavelength of the absorption change of interest allows identification of the pigment responsible from its unique vibrational signature. Using this technique, the origin of ΔA(535) was previously shown to be a subpopulation of red-shifted zeaxanthin molecules. In the absence of zeaxanthin (and antheraxanthin), a proportion of NPQ remains, and the ΔA(535) change is blue-shifted to 525 nm (ΔA(525)). Using resonance Raman spectroscopy, it is shown that the ΔA(525) absorption change in Arabidopsis leaves lacking zeaxanthin belongs to a red-shifted subpopulation of violaxanthin molecules formed during NPQ. The presence of the same ΔA(535) and ΔA(525) Raman signatures in vitro in aggregated LHCII, containing zeaxanthin and violaxanthin, respectively, leads to a new proposal for the origin of the xanthophyll red shifts associated with NPQ.     

Chlorophyll Catabolites in Senescent Leaves of the Lime Tree (Tilia cordata)

In cold extracts of senescent leaves of the Lime tree (Tilia cordata), two colorless nonfluorescent chlorophyll catabolites (NCCs) were identified, named Tc‐NCC‐1 and Tc‐NCC‐2, as well as a polar yellow chlorophyll catabolite (YCC), named Tc‐YCC. The constitution of the two NCCs was determined by spectroscopic means. In addition, a tentative structure was derived for Tc‐YCC. The three chlorophyll degradation products exhibited tetrapyrrolic structures, as are typical of NCCs or YCCs, and turned out to be rather polar, due to a glucopyranosyl group at their 8²‐position. At their 3‐positions, the more polar Tc‐NCC‐1 carried a 1,2‐dihydroxyethyl group and the less polar Tc‐NCC‐2 a vinyl group. Tc‐YCC was identified as the product of an oxidation of Tc‐NCC‐1.     

Differences in photoprotective pigment production between Japanese and Australian strains of Chattonella marina (Raphidophyceae)

Previous studies have shown that isolates of Chattonella marina from Australia and Japan exhibit differences in tolerance to high intensities of visible light. Here we show that the Australian strain of C. marina produces around five times more UV-absorbing mycosporine amino acids (MAAs) than the Japanese strain. This corresponds with 66% increased growth by the Australian strain under UVB exposure compared to no UV exposure. The MAA mycosporine-glycine, which reportedly acts as an antioxidant, was found in high quantity (110 fg cell^-1) in the Australian but was absent in the Japanese strain. In contrast, changes in the concentration of violaxanthin and zeaxanthin per cell were 4.7-4.8 times greater in the Japanese relative to the Australian strain suggesting that the Japanese strain uses a xanthophyll cycle to moderate inhibition by high photosynthetically active radiation (PAR) irradiance. Increased MAA production under high irradiance was also observed in other Australian strains of Chattonella, but not noted in other Japanese strains suggesting ecophenotypic adaptation due to differing environmental conditions.     

Butterfly wing pattern mutants: developmental heterochrony and co-ordinately regulated phenotypes

Butterfly wings are colored late in development, when pigments are synthesized in specialized wing scale cells in a fixed developmental succession. In this succession, colored pigments are deposited first and the remaining areas are later melanized black or brown. Here we studied the developmental changes underlying two wing pattern mutants, firstly melanic mutants of the swallowtail Papilio glaucus, in which the yellow background is turned black, and secondly a Spotty mutant of the satyrid Bicyclus anynana, which carries two additional eyespots. Despite the very different pattern changes in these two mutants, they are both associated with changes in rates of scale development and correspondingly, the final color pattern. In the melanic swallowtail, background scales originally destined to become yellow (normally developing early and synthesizing papiliochrome) show delayed development, fail to make papiliochrome, and subsequently melanize at the same time as scales in the wild-type black pattern. In the B. anynana eyespot, scale maturation begins with the central white focus, then progresses to the surrounding gold ring and later finishes with melanization of the black center. Mutants showing additional eyespots display accelerated rates of scale development (corresponding to new eyespots) in wing cells not normally occupied by eyespots. Thus by either delaying or accelerating rates of scale development, the final color, or position, of a wing pattern element can be changed. We propose that this heterochrony of scale development is a basic mechanism of color pattern formation on which developmental mutants act to change lepidopteran color patterns. Received: 20 April 2000 / Accepted: 19 July 2000     

Effects of iron concentration on pigment composition in Phaeocystis antarctica grown at low irradiance

Interpretation of photosynthetic pigment data using iterative programs such as CHEMTAX are widely used to examine algal community structure in the surface ocean. The accuracy of such programs relies on understanding the effects of environmental parameters on the pigment composition of taxonomically diverse algal groups. Phaeocystis antarctica is an important contributor to total autotrophic production and the biogeochemical cycling of carbon and sulfur in the Southern Ocean. Here we report the results of a laboratory culture experiment in which we examined the effects of ambient dissolved iron concentration on the pigment composition of colonial P. antarctica, using a new P. antarctica strain isolated from the southern Ross Sea in December 2003. Low-iron (<0.2 nM dissolved Fe) filtered Ross Sea seawater was used to prepare the growth media, thus allowing sub-nanomolar iron additions without the use of EDTA to control dissolved iron concentrations. The experiment was conducted at relatively low irradiance (∼20 μE m⁻² s⁻¹), with P. antarctica primarily present in the colonial form—conditions that are typical of the southern Ross Sea during austral spring. Relative to the iron-limited control treatments (0.22 nM dissolved Fe), iron addition mediated a decrease in the ratio of 19′-hexanoyloxyfucoxanthin to chlorophyll a, and an increase in the ratio of fucoxanthin to chlorophyll a. Our results also suggest that the ratio of 19′-hexanoyloxyfucoxanthin to chlorophyll c₃ (Hex:Chl c₃ ratio) may be a characteristic physiological indicator for the iron-nutritional status of colonial P. antarctica, with higher Hex:Chl c₃ ratios (>3) indicative of Fe stress. We also observed that the ratio of fucoxanthin to 19′-hexanoyloxyfucoxanthin (Fuco:Hex ratio) was highly correlated (r ² = 0.82) with initial dissolved Fe concentration, with Fuco:Hex ratios <0.05 measured under iron-limited conditions (dissolved Fe <0.45 nM). Our results corroborate and extend the results of previous experimental studies, and, combined with pigment measurements from the southern Ross Sea, are consistent with the hypothesis that the interconversion of fucoxanthin and 19′-hexanoyloxyfucoxanthin by colonial P. antarctica is used as a photo-protective or light-harvesting mechanism, according to the availability of dissolved iron.