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At least 487 records · Page 27Linked to original sources
PubMed1993

Microamperometric estimation of photosynthesis inhibition in a single algal protoplast.

The single-cell level inhibition of photosynthesis by some chemicals was investigated from the light-irradiation induced response of intracellular oxygen reduction current at an ultramicroelectrode inserted into a single algal protoplast. The addition of 3-(3,4-dichlorophenyl)-1,1-dimethylurea (DCMU), an electron-transport inhibitor, decreased a current peak appeared immediately after the light irradiation. The pI50 value (the negative decadic logarithm of the concentration needed for 50% inhibition) was determined from the change in response patterns and found to be 6.9 for DCMU. 2,4-Dichlorophenoxyacetic acid (2,4-D), an auxin herbicide, showed no effect on photosynthesis in the effective concentration range. 2,4-Dinitrophenol (DNP), a proton carrier, accelerated the oxygen production upon light irradiation.

2,4-Dichlorophenoxyacetic Acid↗
PubMed1997

Chaperonin genes of the Synechocystis PCC 6803 are differentially regulated under light-dark transition during heat stress.

Transcriptional startpoints of the two heat inducible chaperonin genes of Synechocystis PCC 6803 were mapped within the conservative CIRCE element and proved to be identical irrespective of the temperature treatment. Finding of an ORF encoding for a potential CIRCE binding repressor (HrcA) further suggests that both groEL-analogs are regulated in a CIRCE-dependent manner. In contrast to the expectations, the chaperonin twins are differentially expressed under light-dark transition during heat stress. Not the light per se, but rather the photosynthetic electron transport appears to be accountable for the regulatory differences. Our findings support the hypothesis that multiple chaperonins play different physiological roles under stress conditions.

Amino Acid Sequence↗
PubMed1995

Many combinations of amino acid sequences in a conserved region of the D1 protein satisfy photosystem II function.

The putative de helix of the D1 protein is located at the acceptor side of photosystem II (PS II) and serves as an indispensable part of a niche that binds the secondary plastoquinone QB. Combinatorial mutagenesis was applied to a stretch of four residues in a highly conserved region of this putative helix in order to reveal amino acid combinations that are able to support PS II function. An obligate photoheterotrophic mutant of the cyanobacterium Synechocystis sp. PCC 6803, missing four residues (delta YFGR254-7) in the de helix, was transformed with a D1-coding sequence carrying fully degenerate combinations of codons at the site of the deletion. Upon selection for photoautotrophy, 25 mutants with functional PS II were isolated. All mutants showed different codon combinations at positions 254 to 257; none was identical to the wild-type sequence, and none of the conserved residues was found to be mandatory for PS II function. However, 24 of the mutants contained Tyr of Phe at position 254 while at the other three positions many different amino acid combinations could be functionally accommodated. Most sequences maintained an amphiphilic arrangement of the helix that may align Tyr254 facing the QB binding pocket. This residue is proposed to be functionally analogous to Phe216 of the L subunit in purple bacteria which contributes to binding of QB. Most of the PS II properties were similar in the mutants compared to wild-type. Noticeable modifications in the mutants concerned the semiquinone equilibrium of electron transfer between QA and QB, and the affinity of PS II inhibitors. Differential effects on the semiquinone equilibrium were observed between two distinct quinones occupying the QB site (plastoquinone versus 2,5-dichloro-p-benzo- quinone), implying that residues in this domain are involved, directly or indirectly, with different binding determinants of the quinones. Even though many different combinations of amino acids in positions 254 to 257 of the D1 protein may satisfy the primary function of PS II, complex requirements need to be combined for optimized performance of the QB binding niche.

Amino Acid Sequence↗
PubMed2002

Silencing of a gene encoding a protein component of the oxygen-evolving complex of photosystem II enhances virus replication in plants.

It has been suggested that, in addition to viral proteins, host proteins are involved in RNA virus replication. In this study the RNA helicase domain of the Tobacco mosaic virus (TMV) replicase proteins was used as bait in the yeast two-hybrid system to identify tobacco proteins with a putative role in TMV replication. Two host proteins were characterized. One protein (designated #3) belongs to a protein family of ATPases associated with various activities (AAA), while the second host protein (designated #13) is the 33K subunit of the oxygen-evolving complex of photosystem II. Using Tobacco rattle virus vectors, genes #3 and #13 were silenced in Nicotiana benthamiana, after which the plants were challenged by TMV infection. Silencing of gene #13 resulted in a 10-fold increase of TMV accumulation, whereas silencing of gene #3 caused a twofold reduction of TMV accumulation. Additionally, silencing of genes #3 and #13 decreased and increased, respectively, the accumulation of two other viruses. Similar to silencing of gene #13, inhibition of photosystem II by application of an herbicide increased TMV accumulation several fold. Infection of N. benthamiana with TMV resulted in a decrease of #13 mRNA levels. Silencing of gene #13 may reflect a novel strategy of TMV to suppress basal host defense mechanisms. The two-hybrid screenings did not identify tobacco proteins involved in helicase domain-induced N-mediated resistance.

Adenosine Triphosphatases↗
PubMed2001

Characterization and application of plant hairy roots endowed with photosynthetic functions.

The scope of this review includes the physiological properties of the hairy roots induced by light irradiation and the kinetic analysis considering the effects of light intensity on hairy root cultures. The cell lines of photomixotrophic and photoautotrophic hairy roots of pak-bung are established from heterotrophic ones by improving the photosynthetic ability of hairy roots through acclimation cultures under light irradiation. Comparisons of physiological properties of derived photoautotrophic cell line with photomixotrophic and heterotrophic ones are also made through histological examination. Moreover, the effect of photosynthesis inhibitor on the photoautotrophic growth of the hairy roots is described. By elucidating the influences of light intensity on growth and chlorophyll formation of photoautotrophic and photomixotrophic hairy roots, a kinetic model was applied to describe the hairy root growth and chlorophyll formation of these cell lines of hairy roots.

Cell Culture Techniques↗
PubMed1991

Photosynthetic electron transport controls degradation but not production of psbA transcripts in the cyanobacterium Synechocystis 6803.

Accumulation and stability of psbA and rbcL-S transcripts in Synechocystis 6803 was followed in the presence and absence of the photosynthesis inhibitors DCMU and methylviologen. Our results demonstrate that both transcript production and transcript stability are important regulatory elements for psbA gene expression in Synechocystis 6803. The production of psbA transcripts was stimulated by light in a process that operated independently of the photosynthetic electron transport. However, stability of the psbA transcript increased in the dark and was controlled by photosynthetic electron transport. The psbA transcript was remarkably stable in the dark, with a half-life of approximately 7 hours. By contrast, the regulatory pattern for the rbcL-S genes was quite different. The light-stimulated production of rbcL-S transcripts was dependent on an intact photosynthetic electron transport, and rbcL-S transcript stability was higher under illuminated conditions than in darkness.

Blotting, Northern↗
PubMed1996

The Chlamydomonas reinhardtii LI818 gene represents a distant relative of the cabI/II genes that is regulated during the cell cycle and in response to illumination.

In the green unicellular alga Chlamydomonas reinhardtii, as in higher plants, the expression of the genes encoding the chlorophyll a/b-binding (CAB) polypeptides associated with photosystem I (PSI) and photosystem II (PSII) is regulated by endogenous (circadian clock) and exogenous signals (light and temperature). The circadian clock ensures that the oscillation in the levels of the different cab mRNAs is continuously kept in phase with light/dark (LD) cycles and is maximal by the middle of the day. On the other hand, light controls the amplitude of the oscillations. We report here the cloning and characterization of the C. reinhardtii LI818 gene, which identifies a CAB-related polypeptide and whose expression is regulated quite differently from the cab I/II genes. We show: (1) that in LD synchronized Chlamydomonas cells LI818 mRNA accumulation is subject to dual regulation that involves separable regulation by light and an endogenous oscillator; (2) that LI818 mRNA is fully expressed several hours before the cab I/II mRNAs and that the latter accumulate concomitantly; (3) that blocking the electron flow through PSII using DCMU prevents cells from accumulating cab I/II mRNAs but not LI818 mRNA and (4) that the accumulation of LI818 mRNA is abolished by blocking cytoplasmic protein synthesis, suggesting that these regulatory mechanisms are mediated by labile proteins.

Algal Proteins↗
PubMed1996

Site-directed mutagenesis of the CP 47 protein of photosystem II: 167W in the lumenally exposed loop C is required for photosystem II assembly and stability.

The intrinsic chlorophyll-protein CP 47 is a component of photosystem II which functions in both light-harvesting and oxygen evolution. Using site-directed mutagenesis we have produced the mutant W167S which lies in loop C of CP 47. This strain exhibited a 75% loss in oxygen evolution activity and grew extremely slowly in the absence of glucose. Examination of normalized oxygen evolution traces indicated that the mutant was susceptible to photoinactivation. Analysis of the variable fluorescence yield indicated that the mutant accumulated very few functional PS II reaction centers. This was confirmed by immunoblotting experiments. Interestingly, when W167S was grown in the presence of 20 microM DCMU, the mutant continued to exhibit these defects. These results indicate that tryptophan 167 in loop C of CP 47 is important for the assembly and stability of the PS II reaction center.

Cyanobacteria↗
PubMed1993

Functional analysis of the two homologous psbA gene copies in Synechocystis PCC 6714 and PCC 6803.

The cyanobacteria Synechocystis 6803 and 6714 contain three genes (psbA) coding for the D1 protein. This protein is an essential subunit of photosystem II (PSII) and is the target for herbicides. We have used herbicide-resistant mutants to study the role of the two homologous copies of the psbA genes in both strains (the third copy is not expressed). Several herbicide resistance mutations map within the psbAI gene in Synechocystis 6714 (G. Ajlani et al., Plant Mol. Biol. 13 (1989): 469-479). We have looked for mutations in copy II. Results show that in Synechocystis 6714, only psbAI contains herbicide resistance mutations. Relative expression of psbAI and psbAII has been measured by analysing the proportions of resistant and sensitive D1 in the thylakoid membranes of the mutants. In normal growth conditions, 95% resistant D1 and 5% sensitive D1 were found. In high light conditions, expression of psbAII was enhanced, producing 15% sensitive D1. This enhancement is specifically due to high light and not to the decrease of D1 concentration caused by photoinhibition. Copy I of Synechocystis 6714 corresponds to copy 2 of Synechocystis 6803 since it was always psbA2 which was recombined in Synechocystis 6803 transformants. PSII of the transformant strains was found to be 95% resistant to herbicides as in resistant mutants of Synechocystis 6714.

Amino Acid Sequence↗
PubMed1995

Electron transport controls transcription of the glutamine synthetase gene (glnA) from the cyanobacterium Synechocystis sp. PCC 6803.

The glnA gene, encoding type I glutamine synthetase (GS) in Synechocystis sp. PCC 6803, showed a high sequence similarity with other cyanobacterial glnA genes. A dramatic decrease in the amount of glnA mRNA, a single transcript of about 1.6 kb, was observed after transfer to darkness, or after incubation with the electron transport inhibitors DCMU or DBMIB. The levels of glnA transcript were fully recovered after 5 min of reillumination. The glnA mRNA was found to be equally stable both in the light and the dark (half-life about 2.5 min). Unlike the glnA messenger, the amount of GS protein was not reduced in the dark. Synthesis of the glnA transcript in the dark required the presence of glucose. In addition, glnA transcription in a Synechocystis psbE-psbF mutant lacking photosystem II required the presence of glucose even when grown in the light. These observations indicate that glnA transcription is under the control of the redox state of the cell. Finally, nitrogen starvation provoked a delay in the decrease of glnA transcript in darkness, suggesting a connection between nitrogen and redox controls of glnA transcript levels.

Cyanobacteria↗
PubMed1995

Novel aspects of the regulation of a cDNA (Arf1) from Chlamydomonas with high sequence identity to animal ADP-ribosylation factor 1.

ADP-ribosylation factor (ARF) is a highly conserved, low molecular mass (ca. 21 kDa) GTP-binding protein that has been implicated in vesicle trafficking and signal transduction in yeast and mammalian cells. However, little is known of ARF in plant systems. A putative ARF polypeptide was identified in subcellular fractions of the green alga Chlamydomonas reinhardtii, based on [32P]GTP binding and immunoblot assays. A cDNA clone was isolated from Chlamydomonas (Arf1), which encodes a 20.7 kDa protein with 90% identity to human ARF1. Northern blot analyses showed that levels of Arf1 mRNA are highly regulated during 12 h/12 h light/dark (LD) cycles. A biphasic pattern of expression was observed: a transient peak of Arf1 mRNA occurred at the onset of the light period, which was followed ca. 12 h later by a more prominent peak in the early to mid-dark period. When LD-synchronized cells were shifted to continuous darkness, the dark-specific peak of Arf1 mRNA persisted, indicative of a circadian rhythm. The increase in Arf1 mRNA at the beginning of the light period, however, was shown to be light-dependent, and, moreover, dependent on photosynthesis, since it was prevented by DCMU. We conclude that the biphasic pattern of Arf1 mRNA accumulation during LD cycles is due to regulation by two different factors, light (which requires photosynthesis) and the circadian clock. Thus, these studies identify a novel pattern of expression for a GTP-binding protein gene.

ADP-Ribosylation Factor 1↗
PubMed1989

Molecular analysis of psbA mutations responsible for various herbicide resistance phenotypes in Synechocystis 6714.

Mutations conferring herbicide resistance in 3 mutant strains of the cyanobacterium Synechocystis 6714 have been characterized by gene cloning and sequencing. The mutants display very different phenotypes: DCMU-IIA is DCMU-resistant and atrazine-resistant, DCMU-IIB is DCMU-resistant and atrazine-sensitive, and Az-V is DCMU-sensitive, atrazine-resistant and presents particular photoinhibition properties. These mutants were originally obtained either by one-step selection (DCMU-IIA) or by two-step selection (DCMU-IIB and Az-V). psbA copies carrying herbicide resistance have been identified by transformation experiments as psb AI in all cases. Sequences of the psb AI copy of each mutant have been compared to the wild-type sequence. In the single mutant DCMU-IIA, a point mutation at codon 264 (Ser----Ala) results in resistance to both DCMU and atrazine. In the double mutants DCMU-IIB and Az-V, two point mutations were found. DCMU-IIB was derived from DCMU-IIA and had acquired a second mutation at codon 255 (Phe----Leu) resulting in a slight increase in DCMU resistance and complete abolition of atrazine resistance. Az-V contains two changes at codons 211 (Phe----Ser) and 251 (Ala----Val) resulting in high atrazine resistance but only slight DCMU resistance.

Amino Acid Sequence↗
PubMed1990

Light-stimulated cell expansion in bean (Phaseolus vulgaris L.) leaves. I. Growth can occur without photosynthesis.

Cell expansion in dicotyledonous leaves is strongly stimulated by bright white light (WL), at least in part as a result of light-induced acidification of the cell walls. It has been proposed that photosynthetic reactions are required for light-stimulated transport processes across plasma membranes of leaf cells, including proton excretion. The involvement of photosynthesis in growth and wall acidification of primary leaves of bean has been tested by inhibiting photosynthesis in two ways: by reducing chlorophyll content of intact plants with tentoxin (TX) and by treating leaf discs with 3-(3,4-dichlorophenyl)-1,1-dimethylurea (DCMU). Exposure to bright WL stimulated growth of intact leaves of TX-treated plants. Discs excised from green as well as from TX-or DCMU-treated leaves also responded by growing faster in WL, as long as exogenous sucrose was supplied to the photosynthetically inhibited tissues. The WL caused acidification of the epidermal surface of intact TX-leaves, but acidification of the incubation medium by mesophyll cells only occurred when photosynthesis was not inhibited. It is concluded that light-stimulated cell enlargement of bean leaves, and the necessary acidification of epidermal cell walls, are mediated by a pigment other than chlorophyll. Light-induced proton excretion by mesophyll cells, on the other hand, may require both a photosynthetic product (or exogenous sugars) and a non-photosynthetic light effect.

Cell Division↗
PubMed1978

Hydrogen metabolism in the facultative anoxygenic cyanobacteria (blue-green algae) Oscillatoria limnetica and Aphanothece halophytica.

Two facultative anoxygenic photoautotrophic cyanobacteria, Oscillatoria limnetica and Aphanothece halophytica were found capable of CO2 photoassimilation using molecular hydrogen as electron donor in a photosystem I driven reaction. A. halophytica was also capable of evolving hydrogen from Na-dithionite reduced methylviologen in a light independent reaction.

Anaerobiosis↗
PubMed1989

Anaerobic biosynthesis of unsaturated fatty acids in the cyanobacterium, Oscillatoria limnetica.

The mechanism for synthesis of monounsaturated fatty acids under aerobic and anaerobic conditions was studied in the facultative anaerobic cyanobacterium, Oscillatoria limnetica. The hexadecenoic acid (C16:1) of aerobically grown O. limnetica was shown to contain both the delta 7 (79%) and delta 9 (21%) isomers, while the octadecenoic (C18:1) acid was entirely the delta 9 acid. Incorporation of [2-14C] acetate into the fatty acids under aerobic conditions resulted in synthesis of the delta 7 and delta 9 C16:1 and the delta 9 C18:1. Synthesis of unsaturated fatty acids in the presence of DCMU required sulfide. Anaerobic incubations in the presence of DCMU and sulfide (less than 0.003% atmospheric oxygen) resulted in a two-fold increase in monounsaturated fatty acids of both delta 7 and delta 9 C16:1 and delta 9 and delta 11 C18:1. The synthesis of these is characteristic of a bacterial-type, anaerobic pathway.

Acetates↗