Effects of proteinase K on the energy transfer between phycobiliproteins in phycobilisomes

Spectral changes in fluorescence of phycobilisomes (PBS) of A. variabilis treated with proteinase K were studied at room and liquid nitrogen temperature. In control PBS, the relative yield of 77 K fluorescence of F686 was very high, and those of F655 and F666 were low. In PBS treated with proteinase K for less than 1 h, F686 decreased, and F655 and F666 increased. In PBS treated with proteinase K for 2 h, F655 was the main peak of fluorescence emission, F686 was the second peak, the fluorescence emission peak of F666 disappeared. In PBS treated with proteinase K for more than 8 h, F655 showed only one fluorescence emission peak.We suggested that phycobiliporteins in the PBS of A. variabilis constitute an energy transfer chain, shown as follows:{fx91-1}The linkages between APC and APC-B, C-PC and APC, and C-PC and APC-B had different sensitivity towards proteinase K.     

IMMUNOCYTOCHEMICAL CHARACTERIZATION OF THE INTRAPYRENOID THYLAKOIDS OF CRYPTOMONADS1

Thylakoid lamellae extend into the pyrenoids of only two genera of cryptomonad algae, Chroomonas and Hemiselmis, We used immunoelectron microscopy to assess the photosynthetic competency of cryptomonad intrapyrenoid thylakoids. Intrapyrenoid thylakoids possess phycobiliproteins and the chlorophyll a/c₂ light-harvesting complex, both of which are associated with photosystem (PS) II in a light-harvesting capacity. In addition, thylakoids that extend into the pyrenoid of Hemiselmis brunnescens were immunolabelled by anti-PSI. These results indicate that cryptomonad intrapyrenoid thylakoids likely function in a manner analogous to thylakoids of the chloroplast stroma. Moreover, our observation that the Calvin cycle enzyme ribulose 1,5-bisphosphate carboxylase/oxygenase (Rubisco) is pyrenoid-localized in these two cryptophytes indicates that the processes of photosynthetic O₂-evolution and ribulose 1,5-bisphosphate (RuBP) carboxylation/oxygenation are not spatially separated in these algae.     

藻胆蛋白复合物的合成及其分子内能量传递

通过偶联剂3-(2-吡啶联巯基)丙酸N-羟基琥珀亚胺酯(SPDP)及改变配料比, 合成了两个R-藻红蛋白(R-PE)与C-藻蓝蛋白(C-PC)的复合物A和B.利用吸收光谱确定了分子内R-PE与C-PC的摩尔比为6∶1和2∶1. 通过荧光光谱, 观察到能量传递现象, 并计算出能量传递效率为63%和88%.证明分子内能量传递效率很高. 二硫苏糖醇(DTT)还原连接R-PE与C-PC的二硫桥键后, 能量传递被阻断. 这一现象进一步证明复合物中存在分子内能量传递.     

Nitrogen-fixing cyanobacteria as source of phycobiliprotein pigments. Composition and growth performance of ten filamentous heterocystous strains

Ten strains of filamentous, heterocystous nitrogen-fixing blue-green algae (cyanobacteria) were screened for growth performance and tolerance to temperature, pH, irradiance and salinity, together with their potential as producers of phycobiliprotein pigments. Phycobiliproteins typically accounted for about 50% total cell protein, the prevalent type being C-phycocyanin, followed by alloppycocyanin, with levels of 17 and 11% d.wt, respectively, in some strains of Anabaena and Nostoc. C-phycoerythrin was the major pigment in several Nostoc strains, reaching 10% d.wt. Some strains represent, therefore, excellent sources of one or more phycobiliproteins. All strains tolerated an irradiance of ca 2000 μmol photon m^-2 s^-1. Anabaena sp. ATCC 33047 and Nostoc sp. (Albufera) exhibited the widest optimum range of both temperature (30–45 and 25–40 °C) and pH (6.5–9.5 and 6.0–9.0) for growth, the former also showing significant salt tolerance. In an outdoor open system, productivity of cultures of two phycoerythrin-rich strains of Nostoc was over 20 g (d.wt) m^-2 d^-1 during summer. The growth performance of the allophycocyanin-rich Anabaena sp. ATCC 33047 in outdoor semi-continuous culture has been assessed throughout the year. Productivity values under optimized conditions ranged from 9 (winter) to 24 (summer) g (d.wt) m^-2 d^-1.     

Light capture and pigment diversity in marine and freshwater cryptophytes

Phenotypic traits associated with light capture and phylogenetic relationships were characterized in 34 strains of diversely pigmented marine and freshwater cryptophytes. Nuclear SSU and partial LSU rDNA sequence data from 33 of these strains plus an additional 66 strains produced a concatenated rooted maximum likelihood tree that classified the strains into 7 distinct clades. Molecular and phenotypic data together support: (i) the reclassification of Cryptomonas irregularis NIES 698 to the genus Rhodomonas, (ii) revision of phycobiliprotein (PBP) diversity within the genus Hemiselmis to include cryptophyte phycocyanin (Cr‐PC) 569, (iii) the inclusion of previously unidentified strain CCMP 2293 into the genus Falcomonas, even though it contains cryptophyte phycoerythrin 545 (Cr‐PE 545), and (iv) the inclusion of previously unidentified strain CCMP 3175, which contains Cr‐PE 545, in a clade with PC‐containing Chroomonas species. A discriminant analysis‐based model of group membership correctly predicted 70.6% of the clades using three traits: PBP concentration · cell^?1, the wavelength of PBP maximal absorption, and habitat. Non‐PBP pigments (alloxanthin, chl‐a, chl‐c₂, α‐carotene) did not contribute significantly to group classification, indicating the potential plasticity of these pigments and the evolutionary conservation of the PBPs. Pigment data showed evidence of trade‐offs in investments in PBPs vs. chlorophylls (a +c₂).     

Effects of salt stress on basic processes of photosynthesis

Salt stress causes decrease in plant growth and productivity by disrupting physiological processes, especially photosynthesis. The accumulation of intracellular sodium ions at salt stress changes the ratio of K : Na, which seems to affect the bioenergetic processes of photosynthesis. Both multiple inhibitory effects of salt stress on photosynthesis and possible salt stress tolerance mechanisms in cyanobacteria and plants are reviewed.This revised version was published online in March 2005 with corrections to the page numbers.     

Inhibition of Pigments and Phycocolloid in a Marine Red Alga Gracilaria Edulis by Ultraviolet-B Radiation

Vegetative fragments of the subtidal macroalga Gracilaria edulis (Gmel.) Silva cultured under field conditions at Thonithurai were subjected in the laboratory to UV-B radiation (280 – 320 nm). UV-B inhibited the accumulation of chlorophyll and phycobiliproteins, and lowered agar yield (23 to 43%) and its gel strength (22 to 36%) under 12 to 72 h exposure. The longer the exposure to UV-B radiation the more significant impact on pigments and phycocolloid properties of Gracilaria edulis was observed.     

Effects of indole‐3‐butyric acid on growth,pigments and UV‐screening compounds in Nostoc linckia

In the present research, the effect of indole‐3‐butyric acid (IBA) on the growth, and the production of some primary and secondary metabolites was studied in Nostoc linckia. In this respect, algae cultures were supplied with 0, 0.01, 0.1, 1, 10, and 100 μM IBA for 14 days. IBA at concentrations of 10 and 100 μM induced algal growth expressed as fresh weight in N. linckia. Treatment with IBA at all concentrations stimulated heterocyst formation. In addition, low concentrations of IBA (0.01, 0.1, and 1 μM) had a stimulatory effect on chlorophyll a and carotenoids accumulation. In contrast, higher concentrations of IBA induced the accumulation of phycocyanin, allophycocyanin, and phycoerythrin in the treated algae. In this case, IBA at the concentration of 10 μM was more effective. A significant decrease in protein content was observed in the algae treated by 0.01 μM IBA. All concentrations of IBA caused a decrease in sugar content, but lower concentrations were more effective. IBA application in all of the concentrations except 100 μM increased oligosaccharide‐linked mycosporine‐like amino acids (OS‐MAAs) content. Lower concentrations had a more significant effect on increasing OS‐MAAs content. However the concentrations of 10 and 100 μM IBA decreased scytonemin content. These results indicated the stimulatory impact of IBA on weight, heterocyst formation, and photosynthetic pigments in N. linckia.     

Optimization of Maintenance Conditions for Some Microforms of Red Algae in Collections

The effects of light and temperature conditions on the maintenance of some red microalgae (order Porphyridiales) in collections were studied. Periodical subculturing on solid and liquid media was used for maintenance. Halophilic species P. aeruginosa, P. cruentum, and P. sordidum were lost in 2.5 months if kept in darkness. The viability of algae in the dim light slightly declined in 4–5 months and was species-dependent. The results were compared with earlier obtained data on mesophilic Chlorococcales. For the green algae, darkness proved to be the optimal condition, while their viability markedly decreased at light. It was suggested that this discrepancy was caused by the presence of phycobiliproteins in the cells of red algae. Dim light conditions are optimal for the synthesis of phycobiliproteins, which are rapidly destroyed in darkness, thus leading to cell death.     

SYNTHESIS AND BINDING OF PHYCOERYTHRIN AND ITS ASSOCIATED LINKERS TO THE PHYCOBILISOME IN RHODELLA VIOLACEA (RHODOPHYTA): COMPARED EFFECTS OF HIGH LIGHT AND TRANSLATION INHIBITORS1

We studied the synthesis and binding of phycoerythrin and its associated linkers to the phycobilisome (PBS) in Rhodella violacea (Kornmann) Wehrmeyer and compared the effects of high light and translation inhibitors on these processes. Rhodella violacea has a simple hemidiscoidal PBS structure with a well-known composition. The number of PBSs per cell decreases when irradiance is increased, and at higher irradiances the rods are shortened with a specific loss of the terminal hexamer of phycoerythrin (PE) and its associated linker. To test whether or not the observed variations were due to a coordination between the expression of the chloroplast-encoded PE and the nuclear-encoded linkers, we inhibited the expression of the chloroplast genes by the translation inhibitor chloramphenicol. In the few PBSs synthesized, the linker associated to the terminal PE hexamer was missing while that associated with the intermediate PE hexamer was still present. The inhibition by cycloheximide of the translation of the nuclear-encoded linkers did not influence the synthesis of the chloroplast-encoded phycobiliproteins. The absence of linkers prevented the formation of PE hexamers and their binding to the PBSs. We therefore propose the existence of two levels of regulation for PE and associated linkers: the intermediate PE hexamer and associated linker are always present even though their amount is reduced when irradiance is increased. In contrast, the terminal hexamer of PE and its associated linker are no longer present under high light. Their absence can be due to a feedback control between the level of PE and the synthesis of the linker: when the level of PE is lowered below a given value by the action of light on the chloroplast, a signal coming from the chloroplast reaches the nucleus and the synthesis of the linker is repressed. There is no sign of nuclear regulation of the synthesis of PE, but the nuclear-encoded linkers have a structural role in the formation of PE hexamers.