Photosynthesis and apparent affinity for dissolved inorganic carbon by cells and chloroplasts of Chlamydomonas reinhardtii grown at high and low CO2 concentrations

Planta ◽  
1988 ◽  
Vol 176 (2) ◽  
pp. 256-260 ◽  
Author(s):  
Dieter F. S�ltemeyer ◽  
Gerd Kl�ck ◽  
Karlheinz Kreuzberg ◽  
Heinrich P. Fock
2010 ◽  
Vol 37 (8) ◽  
pp. 737 ◽  
Author(s):  
Signe Koch Klavsen ◽  
Stephen C. Maberly

Crassula helmsii (T. Kirk) Cockayne is an invasive aquatic plant in Europe that can suppress many native species because it can grow at a large range of dissolved inorganic carbon concentrations and light levels. One reason for its ecological success may be the possession of a regulated Crassulacean Acid Metabolism (CAM), which allows aquatic macrophytes to take up CO2 in the night in addition to the daytime. The effect of light and CO2 on the regulation of CAM and photosynthesis in C. helmsii was investigated to characterise how physiological acclimation may confer this ecological flexibility. After 3 weeks of growth at high light (230 µmol photon m–2 s–1), C. helmsii displayed 2.8 times higher CAM at low compared with high CO2 (22 v. 230 mmol m–3). CAM was absent in plants grown at low light (23 µmol photon m–2 s–1) at both CO2 concentrations. The observed regulation patterns are consistent with CAM acting as a carbon conserving mechanism. For C. helmsii grown at high light and low CO2, mean photosynthetic rates were relatively high at low concentrations of CO2 and were on average 80 and 102 µmol O2 g–1 DW h–1 at CO2 concentrations of 3 and 22 mmol m–3 CO2, which, together with mean final pH values of 9.01 in the pH drift, indicate a low CO2 compensation point (<3 mmol m–3) but do not indicate use of bicarbonate as an additional source of exogenous inorganic carbon. The relatively high photosynthetic rates during the entire daytime were caused by internally derived CAM-CO2 and uptake from the external medium. During decarboxylation, CO2 generated from CAM contributed up to 29% to photosynthesis, whereas over a day the contribution to the carbon balance was ≤13%. The flexible adjustment of CAM and the ability to maintain photosynthesis at very low external CO2 concentrations, partly by making use of internally generated CO2 via CAM, may contribute to the broad ecological niche of C. helmsii.


2004 ◽  
Vol 31 (10) ◽  
pp. 1027 ◽  
Author(s):  
Xiongwen Chen ◽  
Kunshan Gao

Photosynthetic uptake of inorganic carbon and regulation of photosynthetic CO2 affinity were investigated in Skeletonema costatum (Grev.) Cleve. The pH independence of K1/2(CO2) values indicated that algae grown at either ambient (12 μmol L–1) or low (3 μmol L–1) CO2 predominantly took up CO2 from the medium. The lower pH compensation point (9.12) and insensitivity of photosynthetic rate to di-isothiocyanatostilbene disulfonic acid (DIDS) indicated that the alga had poor capacity for direct HCO3– utilisation. Photosynthetic CO2 affinity is regulated by the concentration of CO2 rather than HCO3–, CO32– or total dissolved inorganic carbon (DIC) in the medium. The response of photosynthetic CO2 affinity to changes in CO2 concentration was most sensitive within the range 3–48 μmol L–1 CO2. Light was required for the induction of photosynthetic CO2 affinity, but not for its repression, when cells were shifted between high (126 μmol L–1) and ambient (12 μmol L–1) CO2. The time needed for cells grown at high CO2 (126 μmol L–1) to fully develop photosynthetic CO2 affinity at ambient CO2 was approximately 2 h, but acclimation to low or very low CO2 levels (3 and 1.3 μmol L–1, respectively) took more than 10 h. Cells grown at low CO2 (3 μmol L–1) required approximately 10 h for repression of all photosynthetic CO2 affinity when transferred to ambient or high CO2 (12 or 126 μmol L–1, respectively), and more than 10 h at very high CO2 (392 μmol L–1).


2008 ◽  
Vol 35 (5) ◽  
pp. 373 ◽  
Author(s):  
Ruby A. Ynalvez ◽  
James V. Moroney

Chlamydomonas reinhardtii is a unicellular eukaryotic alga which possesses a CO2-concentrating mechanism (CCM) that enables it to grow at low CO2 concentrations. Previously, insertional mutants were generated to enable isolation of inorganic carbon transporters and other proteins that might be essential for a functional CCM. These mutants have an antibiotic resistance gene that encodes a protein that binds to Zeocin inhibiting Zeocin’s DNA strand cleavage activity. The DNA flanking the BleR insert of one of the high CO2 requiring strains, named cia7, was cloned with inverse-PCR and sequenced. Sequence analysis showed homology to conserved bacterial proteins of unknown function, but there were no ESTs in this region of the genome. However, the presence of a gene was established by PCR and RLM-RACE. CIA7 was found to have four exons and the BleR insert was in the fourth exon. CIA7 encodes a protein of 104 amino acids with a calculated molecular mass of 11.3 kDa. Based on the ChloroP prediction program, the protein is predicted to have a chloroplast targeting signal. Complementation analyses results showed possible partially rescued mutants, and RNAi showed several transformants with a sick on low CO2 phenotype with reduced expression of CIA7. These results suggest that CIA7 is a gene that facilitates growth in C. reinhardtii under low CO2 conditions. One possible role of CIA7 would be in the delivery or storage of a metal ion. It may play a potential role as either a domain of a metal transporter or as a metallochaperone.


2005 ◽  
Vol 32 (1) ◽  
pp. 55 ◽  
Author(s):  
James E. Adams ◽  
Sergio L. Colombo ◽  
Catherine B. Mason ◽  
Ruby A. Ynalvez ◽  
Baran Tural ◽  
...  

Photosynthetic microorganisms must acclimate to environmental conditions, such as low CO2 environments or high light intensities, which may lead to photo-oxidative stress. In an effort to understand how photosynthetic microorganisms acclimate to these conditions, Chlamydomonas reinhardtii was transformed using the BleR cassette, selected for Zeocin resistance and screened for colonies that showed poor growth at low CO2 levels. One of the insertional mutants obtained, named slc-230, was shown to have a BleR insert in the first exon of Hdh1, a novel, single copy gene. The predicted Hdh1 gene product has similarity to bacterial haloacid dehalogenase-like proteins, a protein family that includes phosphatases and epoxide hydrolases. In addition, Hdh1 is predicted to be localised to the chloroplast or mitochondria in C. reinhardtii. It was found that a genomic copy of wild type Hdh1 can complement slc-230. Physiological studies were conducted to determine the effects of the altered expression of Hdh1 in slc-230. slc-230 grows slowly autotrophically in low CO2, exhibits a lower affinity for inorganic carbon, a decreasing photosynthetic rate over time and a lower content of chlorophylls and quenching xanthophylls than wild type cells. Some possible roles of Hdh1 in the acclimation to low CO2 conditions are discussed.


2010 ◽  
Vol 22 (9) ◽  
pp. 3105-3117 ◽  
Author(s):  
Norikazu Ohnishi ◽  
Bratati Mukherjee ◽  
Tomoki Tsujikawa ◽  
Mari Yanase ◽  
Hirobumi Nakano ◽  
...  

2020 ◽  
Vol 367 (13) ◽  
Author(s):  
Kathleen M Scott ◽  
Tara L Harmer ◽  
Bradford J Gemmell ◽  
Andrew M Kramer ◽  
Markus Sutter ◽  
...  

ABSTRACT Autotrophic microorganisms catalyze the entry of dissolved inorganic carbon (DIC; = CO2 + HCO3− + CO32−) into the biological component of the global carbon cycle, despite dramatic differences in DIC abundance and composition in their sometimes extreme environments. “Cyanobacteria” are known to have CO2 concentrating mechanisms (CCMs) to facilitate growth under low CO2 conditions. These CCMs consist of carboxysomes, containing enzymes ribulose 1,5-bisphosphate oxygenase and carbonic anhydrase, partnered to DIC transporters. CCMs and their DIC transporters have been studied in a handful of other prokaryotes, but it was not known how common CCMs were beyond “Cyanobacteria”. Since it had previously been noted that genes encoding potential transporters were found neighboring carboxysome loci, α-carboxysome loci were gathered from bacterial genomes, and potential transporter genes neighboring these loci are described here. Members of transporter families whose members all transport DIC (CHC, MDT and Sbt) were common in these neighborhoods, as were members of the SulP transporter family, many of which transport DIC. 109 of 115 taxa with carboxysome loci have some form of DIC transporter encoded in their genomes, suggesting that CCMs consisting of carboxysomes and DIC transporters are widespread not only among “Cyanobacteria”, but also among members of “Proteobacteria” and “Actinobacteria”.


1991 ◽  
Vol 69 (5) ◽  
pp. 995-1002 ◽  
Author(s):  
Dieter F. Sültemeyer ◽  
Heinrich P. Fock ◽  
David T. Canvin

Washed protoplasts of low CO2 grown cells of Chlamydomonas reinhardtii were used to further characterize the ability for active CO2 transport. The CO2 transport mechanism and the high affinity for dissolved inorganic carbon were completely induced within 4 h after transferring 5% CO2 grown cells to ambient air (0.035% CO2). Net O2 evolution and CO2 uptake were saturable processes showing saturation between 100 and 200 μM DIC (1.6–3.2 μM CO2) at pH 8.0. For both O2 evolution in whole cells and CO2 uptake in the protoplasts the concentration of dissolved inorganic carbon required for 50% of the maximal rates was about 12 μM (= 0.20 μM CO2). Studies with 3-(3,4-dichloro-phenyl)-1,1 dimethylurea, dibromo-thymoquinone, tetramethyl phenylenediamine and protoplasts of a cytochrome c oxidase deficient mutant of C. reinhardtii indicated the CO2 transport was driven by cyclic or pseudocyclic ATP formation and oxidative phosphorylation was not involved. These studies also show that CO2 transport and CO2 fixation are distinct mechanisms and that active CO2 uptake may occur in the absence of CO2 fixation. Key words: Chlamydomonas reinhardtii; CO2–HCO3− concentrating mechanism, CO2 transport, cyclic photophosphorylation, pseudocyclic photophosphorylation.


2002 ◽  
Vol 29 (3) ◽  
pp. 231 ◽  
Author(s):  
Sergio L. Colombo ◽  
Steve V. Pollock ◽  
Karla A. Eger ◽  
Ashley C. Godfrey ◽  
James E. Adams ◽  
...  

Chlamydomonas reinhardtii Dangeard possesses a CO2 concentrating mechanism (CCM) that enables it to grow at very low CO2 concentrations. In previous studies, insertional mutagenesis was successfully used to identify genes required for growth at low CO2 in C. reinhardtii. These earlier studies used the C. reinhardtii genes, Nit1 and Arg7 to complement nit1– or arg7– strains, thereby randomly inserting a second copy of Nit1 or Arg7 into the genome. Because these genes are already present in the C. reinhardtii genome, it was often difficult to identify the location of the inserted DNA and the gene disrupted by the insertion. We have developed a transformation protocol using the BleR gene, which confers resistance to the antibiotic Zeocin. The insertion of this gene allows one to use a variety of existing polymerase chain reaction (PCR) methodologies to identify the disrupted gene. In this study the D66 strain (nit2–, cw15, mt+) was transformed by electroporation using a plasmid containing the BleR gene. Primary transformants (42 000) were obtained after growth in the dark on acetate plus Zeocin medium. Colonies were then tested for their ability to grow photosynthetically on elevated CO2 or low levels of CO2 (100 ppm). About 120 mutants were identified which grew on elevated CO2 but were unable to grow well at low CO2 concentrations. About 50% of these mutants had low affinities for inorganic carbon as assessed by K0.5(CO2), indicating a potential defect in the CCM. The location of the inserted DNA is being determined using inverse PCR (iPCR) and thermal asymmetric interlaced (TAIL) PCR. Using these methods, one can rapidly locate the inserted DNA in the genome and identify the gene that has been disrupted by the insertion.


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