Iron Deficiency in the Blue-Green Alga Anacystis nidulans: Changes in Pigmentation and Photosynthesis

Phycocyanin-free photosynthetic lamellae (PSI-particles) were prepared from Anacystis nidulans, grown in complete and iron-deficient media. French press treatment and fractionated centrifugation were used. Absorption studies of the particles revealed an iron deficiency-induced shift of the main red chlorophyll a absorption peak from 679 to 673 nm as reported before for whole cells. The shift may reflect a changed distribution between different chlorophyll a forms. Action spectra for photo-oxidation of mammalian cytochrome c with photosynthetic lamellae revealed an iron deficiency-induced shift, corresponding to that found in the absorption spectra. As photo-oxidation of cytochrome c is mediated by PSI, it is believed that chlorophyll a also after the shift towards shorter wavelengths, is active in PSI. A decreased photosynthetic capacity of PSI, due to iron deficiency, was shown by time course studies of photosynthetic oxygen evolution, by photo-oxidation studies of P700 and mammalian cytochrome c, by photo-reduction studies of NADP and by combined studies of light-induced and chemical oxidation of P700. The ration chlorophyll a/700 was also determined for whole cells, lyophilized cells and PSI-particles. Iron deficiency caused an increased ratio in all studied fractions. The results of this work imply that energy is transferred with less efficiency within the photosynthetic units of PSI in iron-deficient A. nidulans than in iron-supplied algae.     

Changes in Pigment Composition and Photosynthesis Induced by Iron-Deficiency in the Blue-Green Alga Anacystis nidulans

Absorption spectra and photosynthetic action spectra have been determined for living Anacystis grown in complete and iron-deficient inorganic media. The absorption studies have shown a spectral shift from 679 nm to 673 nm in the chlorophyll a absorption peak when the algae had to grow without iron. The shift is believed to reflect a changed ratio between at least two chlorophyll a forms denoted Ca670 and Ca680 in this work. Action spectra determinations have revealed a similar shift from 677 nm to 672 nm in the photosynthetic activity peak of chlorophyll a when Anacystis was transferred to a medium without iron. It is proposed that both Ca670 and Ca680 participate in light absorption for photo-system I.     

Ribosomal Ribonucleic Acid Synthesis and Maturation in the Blue-Green Alga Anacystis nidulans

Methods are described for preparation of pulse-labeled ribonucleic acid (RNA) from the blue-green alga Anacystis nidulans. Synthesis of labeled RNA was found to be in part dependent on concurrent photosynthesis and was inhibited by the antibiotic streptolydigin. Mature 23S ribosomal RNA (rRNA) appeared before mature 16S rRNA. Formation of either molecule was inhibited by chloramphenicol, and RNA species of lesser mobility accumulated. These species may be precursors of the mature forms. Maturation of 16S rRNA was also inhibited by streptolydigin. (The effect of this antibiotic on 23S rRNA maturation was not examined). In many respects, ribosomal RNA synthesis and maturation in this blue-green alga appear to follow the pattern already established for bacteria.     

Precursor of 5S Ribosomal Ribonucleic Acid in the Blue-Green Alga Anacystis nidulans

The maturation of 5S ribosomal ribonucleic acid (rRNA) in the obligately photoautotrophic unicellular blue-green alga Anacystis nidulans has been studied by using polyacrylamide gel electrophoresis and T1 ribonuclease oligonucleotide analysis. A. nidulans mature 5S rRNA (m5) is of approximately the same molecular weight as the 5S rRNA of Escherichia coli, and is derived by cleavage of a precursor (p5) containing a few (three to six) additional nucleotides. Some of these additional nucleotides occur at the 5' end of the precursor molecule; others may occur at the 3' end. Kinetic experiments indicate that precursors of mature 5S rRNA larger than p5 either do not exist or are very transient in A. nidulans. These results are discussed in relation to those obtained with other prokaryotes.     

Isolation and Characterization of Ultraviolet Light-Sensitive Mutants of the Blue-Green Alga Anacystis nidulans

Three independently isolated ultraviolet light-sensitive (uvs) mutants of Anacystis nidulans were characterized. Strain uvs-1 was most sensitive to UV in the absence of photoreactivation. Pretreatment with caffeine suppressed the dark-survival curve of strain uvs-1, indicating the presence of excision enzymes involved in dark repair. Under "black" and "white" illumination, strain uvs-1 displays photoreactivation properties nearly comparable to wild-type culture. Mutants uvs-35 and uvs-88 appeared to have partial photorecovery capacities. Upon pretreatment with chloramphenicol, photoreactivation properties of strains uvs-1 and uvs-88 were not evident although the partial photoreactivation characteristics of strain uvs-35 remained the same. Data indicate that strains uvs-1, uvs-35, and uvs-88 are probably genetically distinct UV-sensitive mutants.     

Novel Ribonucleic Acid Species Accumulated in the Dark in the Blue-Green Alga Anacystis nidulans

In the dark, the obligately photoautotrophic blue-green alga Anacystis nidulans accumulates large relative amounts of two novel stable ribonucleic acid species (RNAs). These species are also made in illuminated cells but are unstable in them. When darkened cells are reilluminated, these RNAs are rapidly degraded; degradation is inhibited by chloramphenicol. Upon denaturation with heat or urea, one novel species (0.33 x 10(6) daltons) dissociates into two fragments that comigrate with the second novel species (0.16 x 10(6) daltons) on polyacrylamide gels. Both RNAs are associated with particles sedimenting between 30S and 50S through sucrose gradients and are removed from these particles at low magnesium concentration. The function(s) of these RNAs remains unknown.     

Observations on Replication and Cell Division in Synchronous Cultures of the Blue-Green Alga Anacystis nidulans

A second synchrony induction treatment, half-way through the deoxyribonucleic acid replication process of an already synchronized culture, resulted in defective subsequent cell growth. This supports a previous explanation for the partial loss of viability after synchrony induction.     

Excitation of System I and System II in Photosynthesis as a Possible Control Mechanism of Glycolate Metabolism in the Blue-Green Alga Anacystis nidulans

Photosynthetic enhancement studies performed at 619 nm (excitation of Systems I and II) and at 446 nm (mainly excitation of System I) revealed an 18% photosynthetic enhancement simultaneously with a 31% reduction in glycolate excretion. This observation supports the hypothesis that some glycolate may be consumed in an oxidation process associated with System I when System II is poorly excited and the supply of electrons from the water splitting process of photosynthesis is low.     

Chilling-Susceptibility of the Blue-Green Alga Anacystis nidulans: III. LIPID PHASE OF CYTOPLASMIC MEMBRANE

The lipid phase of cytoplasmic membrane was studied by freeze-fracture electron microscopy in the chilling-susceptible blue-green alga, Anacystis nidulans. At growth temperatures, intramembrane particles were distributed at random in the fracture faces of cytoplasmic membrane, whereas, at chilling temperatures, the fracture faces were composed of particle-free and particle-containing regions. These findings indicate that lipids of the cytoplasmic membrane were in the liquid-crystalline state at the growth temperatures and in the phase-separation state at the chilling temperatures. Temperatures for the onset of phase separation were 5 and 16°C in cells grown at 28 and 38°C, respectively.     

Isolation and Characterization of the Cytoplasmic Membranes from the Blue-Green Alga (Cyanobacterium) Anacystis nidulans

Cells of the blue-green alga (cyanobacterium) Anacyslis nidulanswere disintegrated, and their thylakoid membranes and cytoplasmicmembranes were isolated by floatation centrifugation on a sucrosedensity gradient. Electron micrographs revealed that the cytoplasmicmembranes formed single closed vesicles having diameters of200–400 nm. These membranes contained xanthophylls asthe major constituent pigments and rß-carotene andchlorophyll a as very minor ones. The major peaks in their absorptionspectra were due to carotenoids at 435, 455 and 487 nm, witha minor one due to chlorophyll a at 673 nm. These findings areconsistent with the yellow color of the cytoplasmic membranes.The absorption spectrum of the membranes in the carotenoid regionwas markedly affected by temperature: with a decrease in temperature,the peaks at 455 and 487 nm diminished and a new peak appearedat 390 nm. (Received February 12, 1983; Accepted June 20, 1983)     

Iron Deficiency in the Blue-Green Alga Anacystis nidulans: Fluorescence and Absorption Spectra Recorded at 77°K

Low-temperature (77°K) fluorescence and absorption spectra have been determined for whole cells and photosystem I particles of Anacystis nidulans grown in iron-supplied or iron-deficient inorganic media. Iron deficiency induces a decrease of F720 relative to F685 and F695 in the fluorescence spectra of both whole cells and photosystem I particles. This change is correlated to a reduction of preferentially the long wavelength absorbing fraction of chlorophyll a. The relative fluorescence intensity at 755 nm is increased by iron deficiency. No significant effects of culture-age are found in the ratio between the three fluorescence bands (F685: F695: F720) of iron-supplied A. nidulans.     

Light-Induced Changes in Pigment Composition of Photosynthetic Lamellae and Cell-Free Extracts from the Blue-Green Alga Anacystis nidulans

Anacystis nidulans was grown at two different levels of white light, 7 and50 W^.m^?2. The cells were disrupted through French press treatment, and phycocyanin-free photosynthetic lamellae were obtained from the homogenate by fractionated centrifugation. Comparative absorption studies of the lamellae revealed that high intensity gave an increased carotenoid content relative to chlorophyll a. The spectral characteristics of the cell-free supernatants were also analysed. The high light intensity gave increased contents of both pteridines (410 nm) and allophycocyanin (655 nm) compared with the contents in algae grown at the low light level.     

Changes in Net O(2) Exchange Induced by Inorganic Nitrogen in the Blue-Green Alga Anacystis nidulans

The response of net O₂ exchange to light intensity by intact Anacystis nidulans cells in the presence of saturating NaHCO₃ concentrations followed a curve with an inflection near the light-compensation point. Addition of either KNO₃ or NH₄Cl stimulated O₂ uptake in the dark and at light intensities below the light-compensation point. This resulted in steeper slopes of the curve calculated below and above the light-compensation point. At O₂ concentrations limiting dark respiration, addition of inorganic nitrogen had no effect on either dark respiration or O₂ exchange in the light. The apparent changes in photosynthetic yield observed under normal O₂ concentration disappeared when respiration was limited by O₂ availability, indicating that the effects of inorganic nitrogen on O₂ exchange at low light intensities are due to stimulation of respiration rather than to increases in photosynthetic yield.     

夜间低温对不同光强下生长的两种热带树苗光合作用的影响

于雾凉季测定了叶片叶绿素荧光和气体交换参数、H2 O2 和丙二醛 (MDA)含量 ,探讨了 4～ 6℃夜间低温对 3种相对光强 (太阳光的 8%、2 5 %和 5 0 % )下生长的 2种山地雨林树苗光合作用的影响。夜间低温处理导致 3种光强下生长的 2种植物叶片日间和胁迫诱导的光抑制加剧 ,净光合速率 (Pn)和光呼吸速率降低 ,H2 O2 和MDA含量升高。生长环境光强升高可加剧夜间低温的效应 ,弱光下 2种植物受夜间低温影响较小 ,间接表明雾使光强减弱可缓解自然夜温降低对西双版纳热带植物的危害。处理期间滇南插柚紫Pn 主要受气孔限制 ,滇南红厚壳Pn 降低与活性氧增加引起的羧化效率降低有关 ,两种植物Pn 的降低均与光抑制关系不大。夜间低温后滇南红厚壳气孔导度降低较少 ,H2 O2 和MDA含量较高 ,光抑制较重 ,5 0 %相对光强下其未做夜间低温处理的对照植株发生了胁迫诱导的光抑制 ,表明它对夜间低温较敏感。     

Postmaturational Cleavage of 23s Ribosomal Ribonucleic Acid and Its Metabolic Control in the Blue-Green Alga Anacystis nidulans

Data are presented consistent with the notion that the 23s ribosomal ribonucleic acid (rRNA) of Anacystis nidulans undergoes specific endonucleolytic cleavage in vivo, to produce two fragments with molecular weights of 0.88 x 10(6) and 0.17 x 10(6) daltons. Cleavage occurred at random after 23s rRNA formation and was stimulated by light in this organism, an obligately photoautotrophic unicellular blue-green alga. The half-life of intact 23s rRNA was about 5 h in illuminated cultures and 10 h in unilluminated cultures. 3-(p-Chlorophenyl)-1, 1-dimethylurea, an inhibitor of photosystem II, retarded 23s rRNA cleavage in the light. The results are discussed in the context of recent reports of rRNA instability in a variety of eukaryotic and prokaryotic organisms.     

Primary Role of the Cytoplasmic Membrane in Thermal Acclimation Evidenced in Nitrate-Starved Cells of the Blue-Green Alga, Anacystis nidulans

The lipid phase transition of the cytoplasmic membrane and the chilling susceptibility were studied in nitrate-starved Anacystis nidulans cells. Nitrate starvation resulted in the disappearance of the thylakoid membrane system, without any effect on chilling susceptibility. The chilling susceptibility of the algal cells depended on the growth temperature. Temperatures of lipid phase transitions of the cytoplasmic membranes were detected by chilling-induced spectral changes in the carotenoid region, in vivo. These values were identical to those of cultures containing intact thylakoid systems. Our results suggest that cytoplasmic membrane plays a determinative role in the thermal acclimation of the alga cells.     

Polar Lipids of Chromatium Strain D Grown at Different Light Intensities

The total amount and the composition of polar lipids of the photosynthetic bacterium Chromatium strain D grown in high (7,500 ft/c) and low (100 ft/c) incident white light were measured. Cells grown with low incident light as compared with cells grown in high incident light contained approximately 3.5 times more chlorophyll and contained significantly more internal membranes (chromatophores) as determined by electron microscopy. Nevertheless, there was approximately 40% more phospholipid in cells grown at 7,500 ft/c than in cells grown with 100 ft/c; the phospholipid composition of the cells was similar under both conditions of growth. These observations lead us to suggest that the organization and perhaps localization of the phospholipids in Chromatium may be dependent on the light intensity supplied during growth.     

The Susceptibility of Photosynthesis to Photoinhibition and the Capacity of Recovery in High and Low Light Grown Cyanobacteria, Anacystis nidulans

The susceptibility of photosynthesis to photoinhibition and the rate of its recovery were studied in the cyanobacterium Anacystis nidulans grown at a low (10 micromoles per square meter per second) and a high (120 micromoles per square meter per second) photosynthetically active radiation. The rate of light limited photosynthetic O₂ evolution was measured to determine levels of photoinhibition and rates of recovery. Studies of photoinhibition and recovery with and without the translation inhibitor streptomycin demonstrated the importance of a recovery process for the susceptibility of photosynthesis to photoinhibition. We concluded that the approximately 3 times lower susceptibility to photoinhibition of high light than of low light grown cells, significantly depended on high light grown cells having an approximately 3 times higher recovery capacity than low light grown cells. It is suggested that these differences in susceptibility to photoinhibition and recovery depends on high light grown cells having a higher turnover rate of photosystem II protein(s) that is(are) the primary site(s) of photodamage, than have low light grown cells. Furthermore, we demonstrated that photoinhibition of A. nidulans may occur under physiological light conditions without visible harm to the growth of the cell culture. The results give support for the hypotheses that the net photoinhibitory damage of photosystem II results from the balance between the photoinhibitory process and the operation of a recovery process; the capacity of the latter determining significant differences in the susceptibility of photosynthesis to photoinhibition of high and low light grown A. nidulans.     

pH Changes in the Cytoplasm of the Blue-Green Alga Anacystis nidulans Caused by Light-dependent Proton Flux into the Thylakoid Space

The pH in the cytoplasmic and thylakoid spaces of the blue-green alga, Anacystis nidulans, has been determined in the light and in the dark by uptake of 5,5-dimethyloxazolidine-2,4-dione and methylamine into the sucrose-impermeable ³H-H₂O space, as measured by silicon layer filtering centrifugation.