An efficient protocol for the Agrobacterium-mediated genetic transformation of microalga Chlamydomonas reinhardtii.

Science.gov (United States)

Pratheesh, P T; Vineetha, M; Kurup, G Muraleedhara

2014-06-01

Algal-based recombinant protein production has gained immense interest in recent years. The development of algal expression system was earlier hindered due to the lack of efficient and cost-effective transformation techniques capable of heterologous gene integration and expression. The recent development of Agrobacterium-mediated genetic transformation method is expected to be the ideal solution for these problems. We have developed an efficient protocol for the Agrobacterium-mediated genetic transformation of microalga Chlamydomonas reinhardtii. Pre-treatment of Agrobacterium in TAP induction medium (pH 5.2) containing 100 μM acetosyringone and 1 mM glycine betaine and infection of Chlamydomonas with the induced Agrobacterium greatly improved transformation frequency. This protocol was found to double the number of transgenic events on selection media compared to that of previous reports. PCR was used successfully to amplify fragments of the hpt and GUS genes from transformed cells, while Southern blot confirmed the integration of GUS gene into the genome of C. reinhardtii. RT-PCR, Northern blot and GUS histochemical analyses confirm GUS gene expression in the transgenic cell lines of Chlamydomonas. This protocol provides a quick, efficient, economical and high-frequency transformation method for microalgae.

ChlamyCyc: an integrative systems biology database and web-portal for Chlamydomonas reinhardtii.

Science.gov (United States)

May, Patrick; Christian, Jan-Ole; Kempa, Stefan; Walther, Dirk

2009-05-04

The unicellular green alga Chlamydomonas reinhardtii is an important eukaryotic model organism for the study of photosynthesis and plant growth. In the era of modern high-throughput technologies there is an imperative need to integrate large-scale data sets from high-throughput experimental techniques using computational methods and database resources to provide comprehensive information about the molecular and cellular organization of a single organism. In the framework of the German Systems Biology initiative GoFORSYS, a pathway database and web-portal for Chlamydomonas (ChlamyCyc) was established, which currently features about 250 metabolic pathways with associated genes, enzymes, and compound information. ChlamyCyc was assembled using an integrative approach combining the recently published genome sequence, bioinformatics methods, and experimental data from metabolomics and proteomics experiments. We analyzed and integrated a combination of primary and secondary database resources, such as existing genome annotations from JGI, EST collections, orthology information, and MapMan classification. ChlamyCyc provides a curated and integrated systems biology repository that will enable and assist in systematic studies of fundamental cellular processes in Chlamydomonas. The ChlamyCyc database and web-portal is freely available under http://chlamycyc.mpimp-golm.mpg.de.

Expression of a Synthetic Gene for the Major Cytotoxin (Cyt1Aa of Bacillus thuringiensis subsp. israelensis in the Chloroplast of Wild-Type Chlamydomonas

Directory of Open Access Journals (Sweden)

Seongjoon Kang

2018-05-01

Full Text Available Chlamydomonas reinhardtii (Chlamydomonas strains that are toxic to mosquito larvae because they express chloroplast transgenes that are based on the mosquitocidal proteins of Bacillus thuringiensis subsp. israelensis (Bti could be very useful in mosquito control. Chlamydomonas has several advantages for this approach, including genetic controls not generally available with industrial algae. The Bti toxin is produced by sporulating bacteria and has been used for mosquito control for >30 years without creating highly resistant mosquito populations. The suite of toxins is four main proteins: three Cry proteins and the cytotoxic Cyt1Aa (27 kDa. Cyt1Aa is not very toxic to mosquitoes by itself, but it prevents the development of resistance. The production of Cyt1Aa in other microbes, however, has been challenging due to its affinity for certain membrane phospholipids. Here we report on the production of recombinant Cyt1Aa (rCyt1A in the chloroplast of photosynthetic Chlamydomonas at levels of at least 0.3% total protein. Live cell bioassays demonstrated toxicity of the rCyt1Aa Chlamydomonas to larvae of Aedes aegypti. We also expressed the chloroplast cyt1Aa gene in a wild-type Chlamydomonas strain (21 gr that can grow on nitrate. These results have implications for developing a Chlamydomonas strain that will be toxic to mosquito larvae but will not induce strongly resistant populations.

Metabolic studies of Hg-203 on chlamydomonas reinhardi

International Nuclear Information System (INIS)

Macka, W.; Stehlik, G.; Wihlidal, H.; Washuettl, J.; Bancher, E.

1977-09-01

Vegetative cultures of the green algae Chlamydomonas reinhardi WT + in the log-phase reduce mercury(II)-nitrate to elemental mercury which is removed from the cell suspension by the stream of gas bubbling through it. Monomethyl and dimethyl mercury as intermediate metabolic compounds are to be excluded, because none of them could be found in the algae, the nutrient medium or the gas phase. (author)

ChlamyCyc: an integrative systems biology database and web-portal for Chlamydomonas reinhardtii

Directory of Open Access Journals (Sweden)

Kempa Stefan

2009-05-01

Full Text Available Abstract Background The unicellular green alga Chlamydomonas reinhardtii is an important eukaryotic model organism for the study of photosynthesis and plant growth. In the era of modern high-throughput technologies there is an imperative need to integrate large-scale data sets from high-throughput experimental techniques using computational methods and database resources to provide comprehensive information about the molecular and cellular organization of a single organism. Results In the framework of the German Systems Biology initiative GoFORSYS, a pathway database and web-portal for Chlamydomonas (ChlamyCyc was established, which currently features about 250 metabolic pathways with associated genes, enzymes, and compound information. ChlamyCyc was assembled using an integrative approach combining the recently published genome sequence, bioinformatics methods, and experimental data from metabolomics and proteomics experiments. We analyzed and integrated a combination of primary and secondary database resources, such as existing genome annotations from JGI, EST collections, orthology information, and MapMan classification. Conclusion ChlamyCyc provides a curated and integrated systems biology repository that will enable and assist in systematic studies of fundamental cellular processes in Chlamydomonas. The ChlamyCyc database and web-portal is freely available under http://chlamycyc.mpimp-golm.mpg.de.

The Chlamydomonas genome reveals the evolution of key animal and plant functions

Czech Academy of Sciences Publication Activity Database

Merchant, S.S.; Prochnik, S. E.; Bišová, Kateřina

2007-01-01

Roč. 318, - (2007), s. 245-251 ISSN 0036-8075 Institutional research plan: CEZ:AV0Z50200510 Keywords : chlamydomonas reinhardtii * alga * eukaryotic cell Subject RIV: EE - Microbiology, Virology Impact factor: 26.372, year: 2007

Nonthermal effect of microwave irradiation on nitrite uptake in Chlamydomonas reinhardtii

International Nuclear Information System (INIS)

Pedrajas, C.; Cotrino, J.

1989-01-01

When cells of the unicellular green alga Chlamydomonas reinhardtii were subjected to microwave irradiation at 2.45 GHz, nitrite uptake kinetics still obeyed the Michaelis-Menten equation, the Km of the process remaining constant, whereas V max increased, which indicates an enhanced nonthermal permeability in irradiated cells. (author)

Development of a forward genetic screen to isolate oil mutants in the green microalga Chlamydomonas reinhardtii.

Science.gov (United States)

Cagnon, Caroline; Mirabella, Boris; Nguyen, Hoa Mai; Beyly-Adriano, Audrey; Bouvet, Séverine; Cuiné, Stéphan; Beisson, Fred; Peltier, Gilles; Li-Beisson, Yonghua

2013-12-02

Oils produced by microalgae are precursors to biodiesel. To achieve a profitable production of biodiesel from microalgae, identification of factors governing oil synthesis and turnover is desirable. The green microalga Chlamydomonas reinhardtii is amenable to genetic analyses and has recently emerged as a model to study oil metabolism. However, a detailed method to isolate various types of oil mutants that is adapted to Chlamydomonas has not been reported. We describe here a forward genetic approach to isolate mutants altered in oil synthesis and turnover from C. reinhardtii. It consists of a three-step screening procedure: a primary screen by flow cytometry of Nile red stained transformants grown in 96-deep-well plates under three sequential conditions (presence of nitrogen, then absence of nitrogen, followed by oil remobilization); a confirmation step using Nile red stained biological triplicates; and a validation step consisting of the quantification by thin layer chromatography of oil content of selected strains. Thirty-one mutants were isolated by screening 1,800 transformants generated by random insertional mutagenesis (1.7%). Five showed increased oil accumulation under the nitrogen-replete condition and 13 had altered oil content under nitrogen-depletion. All mutants were affected in oil remobilization. This study demonstrates that various types of oil mutants can be isolated in Chlamydomonas based on the method set-up here, including mutants accumulating oil under optimal biomass growth. The strategy conceived and the protocol set-up should be applicable to other microalgal species such as Nannochloropsis and Chlorella, thus serving as a useful tool in Chlamydomonas oil research and algal biotechnology.

Robust Transgene Expression from Bicistronic mRNA in the Green Alga Chlamydomonas reinhardtii

Directory of Open Access Journals (Sweden)

Masayuki Onishi

2016-12-01

Full Text Available The unicellular green alga Chlamydomonas reinhardtii is a model organism that provides an opportunity to understand the evolution and functional biology of the lineage that includes the land plants, as well as aspects of the fundamental core biology conserved throughout the eukaryotic phylogeny. Although many tools are available to facilitate genetic, molecular biological, biochemical, and cell biological studies in Chlamydomonas, expression of unselected transgenes of interest (GOIs has been challenging. In most methods used previously, the GOI and a selectable marker are expressed from two separate mRNAs, so that their concomitant expression is not guaranteed. In this study, we developed constructs that allow expression of an upstream GOI and downstream selectable marker from a single bicistronic mRNA. Although this approach in other systems has typically required a translation-enhancing element such as an internal ribosome entry site for the downstream marker, we found that a short stretch of unstructured junction sequence was sufficient to obtain adequate expression of the downstream gene, presumably through post-termination reinitiation. With this system, we obtained robust expression of both endogenous and heterologous GOIs, including fluorescent proteins and tagged fusion proteins, in the vast majority of transformants, thus eliminating the need for tedious secondary screening for GOI-expressing transformants. This improved efficiency should greatly facilitate a variety of genetic and cell-biological studies in Chlamydomonas and also enable new applications such as expression-based screens and large-scale production of foreign proteins.

Engineering the chloroplast targeted malarial vaccine antigens in Chlamydomonas starch granules.

Directory of Open Access Journals (Sweden)

David Dauvillée

2010-12-01

Full Text Available Malaria, an Anopheles-borne parasitic disease, remains a major global health problem causing illness and death that disproportionately affects developing countries. Despite the incidence of malaria, which remains one of the most severe infections of human populations, there is no licensed vaccine against this life-threatening disease. In this context, we decided to explore the expression of Plasmodium vaccine antigens fused to the granule bound starch synthase (GBSS, the major protein associated to the starch matrix in all starch-accumulating plants and algae such as Chlamydomonas reinhardtii.We describe the development of genetically engineered starch granules containing plasmodial vaccine candidate antigens produced in the unicellular green algae Chlamydomonas reinhardtii. We show that the C-terminal domains of proteins from the rodent Plasmodium species, Plasmodium berghei Apical Major Antigen AMA1, or Major Surface Protein MSP1 fused to the algal granule bound starch synthase (GBSS are efficiently expressed and bound to the polysaccharide matrix. Mice were either immunized intraperitoneally with the engineered starch particles and Freund adjuvant, or fed with the engineered particles co-delivered with the mucosal adjuvant, and challenged intraperitoneally with a lethal inoculum of P. Berghei. Both experimental strategies led to a significantly reduced parasitemia with an extension of life span including complete cure for intraperitoneal delivery as assessed by negative blood thin smears. In the case of the starch bound P. falciparum GBSS-MSP1 fusion protein, the immune sera or purified immunoglobulin G of mice immunized with the corresponding starch strongly inhibited in vitro the intra-erythrocytic asexual development of the most human deadly plasmodial species.This novel system paves the way for the production of clinically relevant plasmodial antigens as algal starch-based particles designated herein as amylosomes, demonstrating that

Engineering the chloroplast targeted malarial vaccine antigens in Chlamydomonas starch granules.

Science.gov (United States)

Dauvillée, David; Delhaye, Stéphane; Gruyer, Sébastien; Slomianny, Christian; Moretz, Samuel E; d'Hulst, Christophe; Long, Carole A; Ball, Steven G; Tomavo, Stanislas

2010-12-15

Malaria, an Anopheles-borne parasitic disease, remains a major global health problem causing illness and death that disproportionately affects developing countries. Despite the incidence of malaria, which remains one of the most severe infections of human populations, there is no licensed vaccine against this life-threatening disease. In this context, we decided to explore the expression of Plasmodium vaccine antigens fused to the granule bound starch synthase (GBSS), the major protein associated to the starch matrix in all starch-accumulating plants and algae such as Chlamydomonas reinhardtii. We describe the development of genetically engineered starch granules containing plasmodial vaccine candidate antigens produced in the unicellular green algae Chlamydomonas reinhardtii. We show that the C-terminal domains of proteins from the rodent Plasmodium species, Plasmodium berghei Apical Major Antigen AMA1, or Major Surface Protein MSP1 fused to the algal granule bound starch synthase (GBSS) are efficiently expressed and bound to the polysaccharide matrix. Mice were either immunized intraperitoneally with the engineered starch particles and Freund adjuvant, or fed with the engineered particles co-delivered with the mucosal adjuvant, and challenged intraperitoneally with a lethal inoculum of P. Berghei. Both experimental strategies led to a significantly reduced parasitemia with an extension of life span including complete cure for intraperitoneal delivery as assessed by negative blood thin smears. In the case of the starch bound P. falciparum GBSS-MSP1 fusion protein, the immune sera or purified immunoglobulin G of mice immunized with the corresponding starch strongly inhibited in vitro the intra-erythrocytic asexual development of the most human deadly plasmodial species. This novel system paves the way for the production of clinically relevant plasmodial antigens as algal starch-based particles designated herein as amylosomes, demonstrating that efficient production

Evidences of oxidative stress during hydrogen photoproduction in sulfur-deprived cultures of Chlamydomonas reinhardtii

Czech Academy of Sciences Publication Activity Database

Sáens, M. E.; Bišová, Kateřina; Touloupakis, E.; Faraloni, C.; Dario Di Marzio, W.; Torzillo, G.

2015-01-01

Roč. 40, č. 30 (2015), s. 10410-10417 ISSN 0360-3199 Institutional support: RVO:61388971 Keywords : Oxidative stress * Chlamydomonas reinhardtii * H-2 production Subject RIV: EE - Microbiology, Virology Impact factor: 3.205, year: 2015

Toxicity of PAMAM dendrimers to Chlamydomonas reinhardtii

International Nuclear Information System (INIS)

Petit, Anne-Noelle; Eullaffroy, Philippe; Debenest, Timothee; Gagne, Francois

2010-01-01

In recent decades, a new class of polymeric materials, PAMAM dendrimers, has attracted marked interest owing to their unique nanoscopic architecture and their hopeful perspectives in nanomedicine and therapeutics. However, the potential release of dendrimers into the aquatic environment raises the issue about their toxicity on aquatic organisms. Our investigation sought to estimate the toxicity of cationic PAMAM dendrimers on the green alga, Chlamydomonas reinhardtii. Algal cultures were exposed to different concentrations (0.3-10 mg L -1 ) of low dendrimer generations (G2, G4 and G5) for 72 h. Potential adverse effects on Chlamydomonas were assessed using esterase activity (cell viability), photosynthetic O 2 evolution, pigments content and chlorophyll a fluorescence transient. According to the median inhibitory concentration (IC 50 ) appraised from esterase activity, toxicity on cell viability decreased with dendrimer generation number (2, 3 and 5 mg L -1 for G2, G4 and G5 dendrimers, respectively). Moreover, the three generations of dendrimers did not induce the same changes in the photosynthetic metabolism of the green alga. O 2 evolution was stimulated in cultures exposed to the lowest generations tested (i.e. G2 and G4) whereas no significant effects were observed with G5. In addition, total chlorophyll content was increased after G2 treatment at 2.5 mg L -1 . Finally, G2 and G4 had positive effects on photosystem II (PSII): the amount of active PSII reaction centers, the primary charge separation and the electron transport between Q A and Q B were all increased inducing activation of the photosynthetic electron transport chain. These changes resulted in stimulation of full photosynthetic performance.

Comparison of the resistance of two Chlamydomonas reinhardii strains with different β- and carotene content

International Nuclear Information System (INIS)

Gikoshvili, T.I.; Vilenchik, M.M.; Ladygin, V.G.; Kuzin, A.M.

1989-01-01

Radiosensitivity of Chlamydomonas reinhardii strain containing considerable amount of ξ-carotene is lower than that of the wild strain. This indicates that ξ-caotene is oneof the natural radioresistance factors

Toxicity of PAMAM dendrimers to Chlamydomonas reinhardtii

Energy Technology Data Exchange (ETDEWEB)

Petit, Anne-Noelle, E-mail: anne-noelle.petit@ec.gc.ca [Environment Canada, 105 McGill Street, Montreal, Quebec H2Y 2E7 (Canada); Eullaffroy, Philippe [Laboratoire Plantes, Pesticides et Developpement Durable, EA 2069, URVVC, BP 1039, Universite de Reims Champagne-Ardenne, 51687 Reims Cedex 2 (France); Debenest, Timothee; Gagne, Francois [Environment Canada, 105 McGill Street, Montreal, Quebec H2Y 2E7 (Canada)

2010-10-15

In recent decades, a new class of polymeric materials, PAMAM dendrimers, has attracted marked interest owing to their unique nanoscopic architecture and their hopeful perspectives in nanomedicine and therapeutics. However, the potential release of dendrimers into the aquatic environment raises the issue about their toxicity on aquatic organisms. Our investigation sought to estimate the toxicity of cationic PAMAM dendrimers on the green alga, Chlamydomonas reinhardtii. Algal cultures were exposed to different concentrations (0.3-10 mg L{sup -1}) of low dendrimer generations (G2, G4 and G5) for 72 h. Potential adverse effects on Chlamydomonas were assessed using esterase activity (cell viability), photosynthetic O{sub 2} evolution, pigments content and chlorophyll a fluorescence transient. According to the median inhibitory concentration (IC{sub 50}) appraised from esterase activity, toxicity on cell viability decreased with dendrimer generation number (2, 3 and 5 mg L{sup -1} for G2, G4 and G5 dendrimers, respectively). Moreover, the three generations of dendrimers did not induce the same changes in the photosynthetic metabolism of the green alga. O{sub 2} evolution was stimulated in cultures exposed to the lowest generations tested (i.e. G2 and G4) whereas no significant effects were observed with G5. In addition, total chlorophyll content was increased after G2 treatment at 2.5 mg L{sup -1}. Finally, G2 and G4 had positive effects on photosystem II (PSII): the amount of active PSII reaction centers, the primary charge separation and the electron transport between Q{sub A} and Q{sub B} were all increased inducing activation of the photosynthetic electron transport chain. These changes resulted in stimulation of full photosynthetic performance.

Biofiksasi CO2 Oleh Mikroalga Chlamydomonas sp dalam Photobioreaktor Tubular

Directory of Open Access Journals (Sweden)

Hadiyanto Hadiyanto

2014-05-01

Full Text Available Mikroalga memiliki potensi dalam membiofiksasi CO2 dan dapat dimanfaatkan untuk mengurangi kadar CO2 dalam gas pencemar. Pertumbuhan mikroalga sangat dipengaruhi oleh konsentrasi gas CO2 di dalam gas pencemar. Tujuan penelitian ini adalah untuk mengeetahui kemampuan mikroalga Chlamydomonas sp yang dikultivasi dalam photobioreaktor tubular dalam penyerapan gas CO2 serta untuk mengetahui konsentrasi maksimum gas CO2 dalam umpan untuk memproduksi biomasa mikroalga yang optimal. Percobaan dilakukan dnegan memvariasi laju alir dari 0.03 -0.071 L/menit dan konsentrasi CO2 dalam umpan 10-30%. Hasil penelitian menunjukkan bahwa biomasa mikroalga dapat diproduksi dengan maksimal dengan konsentrasi gas CO2 20% dengan laju alir 0.07 L/min. Semakin tinggi laju alir maka produksi biomasa alga semakin besar. Kecepatan pertumbuhan alga maksimum terjadi pada 0.31 /hari. Pada konsentrasi gas CO2 30%, terjadi substrate inhibition yang disebabkan carbon dalam bentuk ion bicarbonate tidak dapat dikonsumsi lagi di dalam kultur alga. Kata kunci : Mikroalga, chlamydomonas sp, biofiksasi CO2, biogas Abstract Microalgae have a potential for CO2 biofixation and therefore can be used to reduce the CO2 concentration in the gas pollutants. Moreover, microalgae growth is strongly affected by the concentration of CO2 in the exhaust gas pollutants. The objective of this research was to investigate the ability of microalgae Chlamydomonas sp which was cultivated in a tubular photobioreactor for CO2 absorption as well as to determine the maximum concentration of CO2 in the feed gas to obtain optimum microalgae biomass. The experiments were performed by varying the gas flow rate of 0.03 -0.071 L / min and the concentration of CO2 in the feed of 10-30%. The results showed that the maximum biomass of microalgae can be produced with CO2 concentration of 20% vol with a flow rate of 0.07 L / min. The result also showed that increasing the gas flow rate, the greater of the production of

Bioenergetics of growth and lipid production in Chlamydomonas reinhardtii

International Nuclear Information System (INIS)

Küçük, Kübra; Tevatia, Rahul; Sorgüven, Esra; Demirel, Yaşar; Özilgen, Mustafa

2015-01-01

The study of thermodynamic aspects of the lipid, e.g., raw material for biodiesel, production in microalgae is important, as the non-lipid producing biological activities of the algal cultivation consume part of the solar energy captured during photosynthesis in expense of the exergetic efficiency of the lipid production process. The cultivation of Chlamydomonas reinhardtii (a unicellular biflagellate fresh-water microalga) is modeled as a three-step chemical mechanism representing growth, respiration, and lipid production. Further, the comprehensive thermodynamic analysis of these mechanisms is presented. The cumulative degree of perfection of the cellular proliferation, after excluding the lipid synthesis, fluctuates with no trend around 0.52 ± 0.19. The exergy analysis has indicated that C. reinhardtii prefers to maximize the lipid production when it is difficult to generate new cells. Under batch production of algal biomass, the highest heat and exergy loss per unit biomass production are accountable under the most favorable biological growth conditions, whereas the highest exergetic efficiency of the lipid production accounted under the least favorable growth conditions, which is in line with the previous studies. - Highlights: • Biomass, lipid production and respiration modeled as three-step chemical reaction. • CDP (cumulative degree of perfection) is calculated based on the model. • The CDP of the algae, after excluding the lipids, is about 0.52 ± 0.19. • Chlamydomonas reinhardtii maximized lipid production when it was difficult to grow

Triclosan-induced transcriptional and biochemical alterations in the freshwater green algae Chlamydomonas reinhardtii

NARCIS (Netherlands)

Pan, Chang Gui; Peng, Feng-Jiao; Shi, Wen Jun; Hu, Li Xin; Wei, Xiao Dong; Ying, Guang Guo

2018-01-01

Triclosan (TCS) is an antibacterial and antifungal agent widely used in personal care products (PCPs). We investigated the effects of TCS (20 μg/L, 100 μg/L and 500 μg/L) on Chlamydomonas reinhardtii by measuring the algal growth, chlorophyll content, lipid peroxidation, and transcription of the

Trophic transfer of gold nanoparticles from Euglena gracilis or Chlamydomonas reinhardtii to Daphnia magna

International Nuclear Information System (INIS)

Lee, Woo-Mi; Yoon, Sung-Ji; Shin, Yu-Jin; An, Youn-Joo

2015-01-01

Understanding the trophic transfer of nanoparticles (NPs) is important because NPs are small enough to easily penetrate into organisms. In this study, we evaluated the trophic transfer of gold NPs (AuNPs) within the aquatic food chain. We observed AuNPs transfer from 2 species of primary producers (Chlamydomonas reinhardtii or Euglena gracilis) to the primary consumer (Daphnia magna). Also, bioaccumulation of AuNPs in E. gracilis was higher than that in C. reinhardtii. The reasons for the difference in Au accumulation may be the physical structure of these organisms, and the surface area that is available for interaction with NPs. C. reinhardtii has a cell wall that may act as a barrier to the penetration of NPs. The size of E. gracilis is larger than that of C. reinhardtii. This study demonstrates the trophic transfer of AuNPs from a general producer to a consumer in an aquatic environment. - Highlights: • This study evaluated the trophic transfer of AuNPs in an aquatic food chain. • Chlamydomonas reinhardtii and Euglena gracilis were selected as the primary producers. • Daphnia magna was used as the primary consumer. • The bioaccumulation of AuNPs in E. gracilis was higher than that in C. reinhardtii. • AuNPs were transferred from C. reinhardtii and E. gracilis to D. magna. - Gold nanoparticles can transfer from primary producers (Chlamydomonas reinhardtii or Euglena gracilis) to the primary consumer (Daphnia magna) in an aquatic environment

Gene silencing of stearoyl-ACP desaturase enhances the stearic acid content in Chlamydomonas reinhardtii

NARCIS (Netherlands)

Jaeger, de L.; Springer, J.; Wolbert, E.J.H.; Martens, D.E.; Eggink, G.; Wijffels, R.H.

2017-01-01

In this study, stearoyl-ACP desaturase (SAD), the enzyme that converts stearic acid into oleic acid, is silenced by artificial microRNA in the green microalga Chlamydomonas reinhardtii. Two different constructs, which target different positions on the mRNA of stearoyl-ACP desaturase, were tested.

Protocol: methodology for chromatin immunoprecipitation (ChIP in Chlamydomonas reinhardtii

Directory of Open Access Journals (Sweden)

Strenkert Daniela

2011-11-01

Full Text Available Abstract We report on a detailed chromatin immunoprecipitation (ChIP protocol for the unicellular green alga Chlamydomonas reinhardtii. The protocol is suitable for the analysis of nucleosome occupancy, histone modifications and transcription factor binding sites at the level of mononucleosomes for targeted and genome-wide studies. We describe the optimization of conditions for crosslinking, chromatin fragmentation and antibody titer determination and provide recommendations and an example for the normalization of ChIP results as determined by real-time PCR.

Biofixation of Carbon dioxide by Chlamydomonas sp. in a Tubular Photobioreactor

Directory of Open Access Journals (Sweden)

H Hadiyanto

2012-02-01

Full Text Available The biogas production from anaerobic digestion is a potential fuel for power generators application, if biogas can be upgraded to the same standards as fossil natural gas by CO2, H2S, and other non-combustible component removal. Microalgae Chlamydomonas sp. has potency to biofix the carbon dioxide and can be used as an additional food ingredient. The variations of flow rate and carbon dioxide concentration in the process resulting different value of biomass production and carbon dioxide biofixation. Biomass production at 40% carbon dioxide concentration obtained 5.685 gr/dm3 at 10% carbon dioxide concentration obtained 4.892 gr/dm3. The greatest value of carbon dioxide absorption occurs at a 40% concentration amounting to 12.09%. The rate of growth and productivity of microalgae tend to rise in 10% and 20% (%v carbon dioxide concentration, but began started a constant at 30% and 40% (%v carbon dioxide concentration. Biomass production tends to increase in light conditions while a constant in dark conditions. This study used Chlamydomonas sp. as media culture and performed on bubble column and tubular reactor with 6 litres of culture medium at a temperature of 28oC and atmospheric pressure.

Comparative genomics in Chlamydomonas and Plasmodium identifies an ancient nuclear envelope protein family essential for sexual reproduction in protists, fungi, plants, and vertebrates.

Science.gov (United States)

Ning, Jue; Otto, Thomas D; Pfander, Claudia; Schwach, Frank; Brochet, Mathieu; Bushell, Ellen; Goulding, David; Sanders, Mandy; Lefebvre, Paul A; Pei, Jimin; Grishin, Nick V; Vanderlaan, Gary; Billker, Oliver; Snell, William J

2013-05-15

Fertilization is a crucial yet poorly characterized event in eukaryotes. Our previous discovery that the broadly conserved protein HAP2 (GCS1) functioned in gamete membrane fusion in the unicellular green alga Chlamydomonas and the malaria pathogen Plasmodium led us to exploit the rare biological phenomenon of isogamy in Chlamydomonas in a comparative transcriptomics strategy to uncover additional conserved sexual reproduction genes. All previously identified Chlamydomonas fertilization-essential genes fell into related clusters based on their expression patterns. Out of several conserved genes in a minus gamete cluster, we focused on Cre06.g280600, an ortholog of the fertilization-related Arabidopsis GEX1. Gene disruption, cell biological, and immunolocalization studies show that CrGEX1 functions in nuclear fusion in Chlamydomonas. Moreover, CrGEX1 and its Plasmodium ortholog, PBANKA_113980, are essential for production of viable meiotic progeny in both organisms and thus for mosquito transmission of malaria. Remarkably, we discovered that the genes are members of a large, previously unrecognized family whose first-characterized member, KAR5, is essential for nuclear fusion during yeast sexual reproduction. Our comparative transcriptomics approach provides a new resource for studying sexual development and demonstrates that exploiting the data can lead to the discovery of novel biology that is conserved across distant taxa.

ChlamyCyc - a comprehensive database and web-portal centered on _Chlamydomonas reinhardtii_

OpenAIRE

Jan-Ole Christian; Patrick May; Stefan Kempa; Dirk Walther

2009-01-01

*Background* - The unicellular green alga _Chlamydomonas reinhardtii_ is an important eukaryotic model organism for the study of photosynthesis and growth, as well as flagella development and other cellular processes. In the era of high-throughput technologies there is an imperative need to integrate large-scale data sets from high-throughput experimental techniques using computational methods and database resources to provide comprehensive information about the whole cellular system of a sin...

Tracking the elusive 5' exonuclease activity of Chlamydomonas reinhardtii RNase J.

Science.gov (United States)

Liponska, Anna; Jamalli, Ailar; Kuras, Richard; Suay, Loreto; Garbe, Enrico; Wollman, Francis-André; Laalami, Soumaya; Putzer, Harald

2018-04-01

Chlamydomonas RNase J is the first member of this enzyme family that has endo- but no intrinsic 5' exoribonucleolytic activity. This questions its proposed role in chloroplast mRNA maturation. RNA maturation and stability in the chloroplast are controlled by nuclear-encoded ribonucleases and RNA binding proteins. Notably, mRNA 5' end maturation is thought to be achieved by the combined action of a 5' exoribonuclease and specific pentatricopeptide repeat proteins (PPR) that block the progression of the nuclease. In Arabidopsis the 5' exo- and endoribonuclease RNase J has been implicated in this process. Here, we verified the chloroplast localization of the orthologous Chlamydomonas (Cr) RNase J and studied its activity, both in vitro and in vivo in a heterologous B. subtilis system. Our data show that Cr RNase J has endo- but no significant intrinsic 5' exonuclease activity that would be compatible with its proposed role in mRNA maturation. This is the first example of an RNase J ortholog that does not possess a 5' exonuclease activity. A yeast two-hybrid screen revealed a number of potential interaction partners but three of the most promising candidates tested, failed to induce the latent exonuclease activity of Cr RNase J. We still favor the hypothesis that Cr RNase J plays an important role in RNA metabolism, but our findings suggest that it rather acts as an endoribonuclease in the chloroplast.

Relation between hydrogen production and photosynthesis in the green algae Chlamydomonas reinhardtii

OpenAIRE

Basu, Alex

2015-01-01

The modernized world is over-consuming low-cost energy sources that strongly contributes to pollution and environmental stress. As a consequence, the interest for environmentally friendly alternatives has increased immensely. One such alternative is the use of solar energy and water as a raw material to produce biohydrogen through the process of photosynthetic water splitting. In this work, the relation between H2-production and photosynthesis in the green algae Chlamydomonas reinhardtii was ...

Refactoring the six-gene photosystem II core in the chloroplast of the green algae Chlamydomonas reinhardtii

DEFF Research Database (Denmark)

Gimpel, Javier A.; Nour-Eldin, Hussam Hassan; Scranton, Melissa A.

2016-01-01

production, particularly under specific environmental conditions. PSII is a complex multisubunit enzyme with strong interdependence among its components. In this work, we have deleted the six core genes of PSII in the eukaryotic alga Chlamydomonas reinhardtii and refactored them in a single DNA construct...

Reverse genetics in Chlamydomonas: a platform for isolating insertional mutants

Directory of Open Access Journals (Sweden)

de Montaigu Amaury

2011-07-01

Full Text Available Abstract A method was developed to identify insertional mutants of Chlamydomonas reinhardtii disrupted for selected target genes. The approach relies on the generation of thousands of transformants followed by PCR-based screenings that allow for identification of strains harboring the introduced marker gene within specific genes of interest. Our results highlight the strengths and limitations of two independent screens that differed in the nature of the marker DNA used (PCR-amplified fragment containing the plasmid-free marker versus entire linearized plasmid with the marker and in the strategies used to maintain and store transformants.

ChlamyCyc: an integrative systems biology database and web-portal for Chlamydomonas reinhardtii

OpenAIRE

May, P.; Christian, J.O.; Kempa, S.; Walther, D.

2009-01-01

Abstract Background The unicellular green alga Chlamydomonas reinhardtii is an important eukaryotic model organism for the study of photosynthesis and plant growth. In the era of modern high-throughput technologies there is an imperative need to integrate large-scale data sets from high-throughput experimental techniques using computational methods and database resources to provide comprehensive information about the molecular and cellular organization of a single organism. Results In the fra...

Characterization of Chlamydomonas reinhardtii Core Histones by Top-Down Mass Spectrometry Reveals Unique Algae-Specific Variants and Post-Translational Modifications.

Science.gov (United States)

Khan, Aliyya; Eikani, Carlo K; Khan, Hana; Iavarone, Anthony T; Pesavento, James J

2018-01-05

The unicellular microalga Chlamydomonas reinhardtii has played an instrumental role in the development of many new fields (bioproducts, biofuels, etc.) as well as the advancement of basic science (photosynthetic apparati, flagellar function, etc.). Chlamydomonas' versatility ultimately derives from the genes encoded in its genome and the way that the expression of these genes is regulated, which is largely influenced by a family of DNA binding proteins called histones. We characterize C. reinhardtii core histones, both variants and their post-translational modifications, by chromatographic separation, followed by top-down mass spectrometry (TDMS). Because TDMS has not been previously used to study Chlamydomonas proteins, we show rampant artifactual protein oxidation using established nuclei purification and histone extraction methods. After addressing oxidation, both histones H3 and H4 are found to each have a single polypeptide sequence that is minimally acetylated and methylated. Surprisingly, we uncover a novel monomethylation at lysine 79 on histone H4 present on all observed molecules. Histone H2B and H2A are found to have two and three variants, respectively, and both are minimally modified. This study provides an updated assessment of the core histone proteins in the green alga C. reinhardtii by top-down mass spectrometry and lays the foundation for further investigation of these essential proteins.

Oil accumulation in the model green alga Chlamydomonas reinhardtii: characterization, variability between common laboratory strains and relationship with starch reserves

Directory of Open Access Journals (Sweden)

Carrier Patrick

2011-01-01

Full Text Available Abstract Background When cultivated under stress conditions, many microalgae species accumulate both starch and oil (triacylglycerols. The model green microalga Chlamydomonas reinhardtii has recently emerged as a model to test genetic engineering or cultivation strategies aiming at increasing lipid yields for biodiesel production. Blocking starch synthesis has been suggested as a way to boost oil accumulation. Here, we characterize the triacylglycerol (TAG accumulation process in Chlamydomonas and quantify TAGs in various wild-type and starchless strains. Results In response to nitrogen deficiency, Chlamydomonas reinhardtii produced TAGs enriched in palmitic, oleic and linoleic acids that accumulated in oil-bodies. Oil synthesis was maximal between 2 and 3 days following nitrogen depletion and reached a plateau around day 5. In the first 48 hours of oil deposition, a ~80% reduction in the major plastidial membrane lipids occurred. Upon nitrogen re-supply, mobilization of TAGs started after starch degradation but was completed within 24 hours. Comparison of oil content in five common laboratory strains (CC124, CC125, cw15, CC1690 and 11-32A revealed a high variability, from 2 μg TAG per million cell in CC124 to 11 μg in 11-32A. Quantification of TAGs on a cell basis in three mutants affected in starch synthesis (cw15sta1-2, cw15sta6 and cw15sta7-1 showed that blocking starch synthesis did not result in TAG over-accumulation compared to their direct progenitor, the arginine auxotroph strain 330. Moreover, no significant correlation was found between cellular oil and starch levels among the twenty wild-type, mutants and complemented strains tested. By contrast, cellular oil content was found to increase steeply with salt concentration in the growth medium. At 100 mM NaCl, oil level similar to nitrogen depletion conditions could be reached in CC124 strain. Conclusion A reference basis for future genetic studies of oil metabolism in Chlamydomonas

Chlamydomonas Outer Arm Dynein Alters Conformation in Response to Ca2+

OpenAIRE

Sakato, Miho; Sakakibara, Hitoshi; King, Stephen M.

2007-01-01

We have previously shown that Ca2+ directly activates ATP-sensitive microtubule binding by a Chlamydomonas outer arm dynein subparticle containing the β and γ heavy chains (HCs). The γ HC–associated LC4 light chain is a member of the calmodulin family and binds 1-2 Ca2+ with KCa = 3 × 10−5 M in vitro, suggesting it may act as a Ca2+ sensor for outer arm dynein. Here we investigate interactions between the LC4 light chain and γ HC. Two IQ consensus motifs for binding calmodulin-like proteins a...

Enhanced methane production of Chlorella vulgaris and Chlamydomonas reinhardtii by hydrolytic enzymes addition

International Nuclear Information System (INIS)

Mahdy, Ahmed; Mendez, Lara; Ballesteros, Mercedes; González-Fernández, Cristina

2014-01-01

Highlights: • Methane production of microalgae biomass is hampered by their cell wall. • Pretreatment should be designed in accordance to the microalgae specie. • Fresh Chlamydomonas reinhardtii exhibited high anaerobic biodegradability. • Chlorella vulgaris anaerobic biodegradability was enhanced by 50% using protease pretreatment. - Abstract: The effect of enzymatic hydrolysis on microalgae organic matter solubilisation and methane production was investigated in this study. Even though both biomasses, Chlamydomonas reinhardtii and Chlorella vulgaris, exhibited similar macromolecular distribution, their cell wall composition provided different behaviors. The addition of carbohydrolase (Viscozyme) and protease (Alcalase) resulted in high carbohydrates and protein solubilisation on both biomasses (86–96%). Despite the high carbohydrate solubilisation with the carbohydrolase, methane production was enhanced by 14% for C. vulgaris, while hydrolyzed C. reinhardtii did not show any improvement. The addition of protease to C. reinhardtii increased methane production by 1.17-fold. The low enhancement achieved together with the inherent high biodegradability of this biomass would not justify the cost associated to the enzyme addition. On the other hand, C. vulgaris hydrolyzed with the protease resulted in 86% anaerobic biodegradability compared to 54% of the raw biomass. Therefore, the application of protease prior anaerobic digestion of C. vulgaris could be a promising approach to decrease the energetic input required for cell wall disruption

Filling Knowledge Gaps in Biological Networks: integrating global approaches to understand H2 metabolism in Chlamydomonas reinhardtii - Final Report

Energy Technology Data Exchange (ETDEWEB)

Posewitz, Matthew C

2011-06-30

The green alga Chlamydomonas reinhardtii (Chlamydomonas) has numerous genes encoding enzymes that function in fermentative pathways. Among these genes, are the [FeFe]-hydrogenases, pyruvate formate lyase, pyruvate ferredoxin oxidoreductase, acetate kinase, and phosphotransacetylase. We have systematically undertaken a series of targeted mutagenesis approaches to disrupt each of these key genes and omics techniques to characterize alterations in metabolic flux. Funds from DE-FG02-07ER64423 were specifically leveraged to generate mutants with disruptions in the genes encoding the [FeFe]-hydrogenases HYDA1 and HYDA2, pyruvate formate lyase (PFL1), and in bifunctional alcohol/aldehyde alcohol dehydrogenase (ADH1). Additionally funds were used to conduct global transcript profiling experiments of wildtype Chlamydomonas cells, as well as of the hydEF-1 mutant, which is unable to make H2 due to a lesion in the [FeFe]-hydrogenase biosynthetic pathway. In the wildtype cells, formate, acetate and ethanol are the dominant fermentation products with traces of CO2 and H2 also being produced. In the hydEF-1 mutant, succinate production is increased to offset the loss of protons as a terminal electron acceptor. In the pfl-1 mutant, lactate offsets the loss of formate production, and in the adh1-1 mutant glycerol is made instead of ethanol. To further probe the system, we generated a double mutant (pfl1-1 adh1) that is unable to synthesize both formate and ethanol. This strain, like the pfl1 mutants, secreted lactate, but also exhibited a significant increase in the levels of extracellular glycerol, acetate, and intracellular reduced sugars, and a decline in dark, fermentative H2 production. Whereas wild-type Chlamydomonas fermentation primarily produces formate and ethanol, the double mutant performs a complete rerouting of the glycolytic carbon to lactate and glycerol. Lastly, transcriptome data have been analysed for both the wildtype and hydEF-1, that correlate with our

Modifying effect of caffeine on lethality and mutability of Chlamydomonas reinhardii cells following UV irradiation

International Nuclear Information System (INIS)

Podstavkova, S.; Vlcek, D.; Miadokova, E.

1983-01-01

The modifying effect of caffeine was studied using two standard and two UV-sensitive strains of Chlamydomonas reinhardii Dang. Cell survival and mutation frequency was microscopically evaluated on media without caffeine and on media with 1.5 mM of caffeine. The obtained results were indicative of the stimulating effect of caffeine upon survival in all strains. (author)

Carbon allocation and element composition in four Chlamydomonas mutants defective in genes related to the CO2 concentrating mechanism

Czech Academy of Sciences Publication Activity Database

Memmola, F.; Mukherjee, B.; Moroney, James V.; Giordano, Mario

2014-01-01

Roč. 121, 2-3 (2014), s. 201-211 ISSN 0166-8595 Institutional support: RVO:61388971 Keywords : Chlamydomonas mutants * carbon * carbon dioxide * elemental stoichiometry Subject RIV: EE - Microbiology, Virology Impact factor: 3.502, year: 2014

Sulphate, more than a nutrient, protects the microalga Chlamydomonas moewusii from cadmium toxicity

Energy Technology Data Exchange (ETDEWEB)

Mera, Roi; Torres, Enrique, E-mail: torres@udc.es; Abalde, Julio

2014-03-01

Highlights: • Sulphate effect on cadmium toxicity in the microalga Chlamydomonas moewusii Gerloff. • Cadmium increases the sulphur requirements in Chlamydomonas moewusii. • Kinetic coefficients for sulphate utilization and cadmium effect on them. • Sulphate and cadmium influence on the biosynthesis of low-molecular mass thiols. • Cadmium toxicity reduction by sulphate due to higher biosynthesis of thiols. - Abstract: Sulphur is an essential macroelement that plays important roles in living organisms. The thiol rich sulphur compounds, such as cysteine, γ-Glu–Cys, glutathione and phytochelatins participate in the tolerance mechanisms against cadmium toxicity. Plants, algae, yeasts and most prokaryotes cover their demand for reduced sulphur by reduction of inorganic sulphate. The aim of this study was to investigate, using a bifactorial experimental design, the effect of different sulphate concentrations in the nutrient solution on cadmium toxicity in the freshwater microalga Chlamydomonas moewusii. Cell growth, kinetic parameters of sulphate utilization and intracellular concentrations of low-molecular mass thiol compounds were determined. A mathematical model to describe the growth of this microalga based on the effects of sulphate and cadmium was obtained. An ANOVA revealed an interaction between them, 16% of the effect sizes was explained by this interaction. A higher amount of sulphate in the culture medium allowed a higher cadmium tolerance due to an increase in the thiol compound biosynthesis. The amount of low-molecular mass thiol compounds, mainly phytochelatins, synthesized by this microalga was significantly dependent on the sulphate and cadmium concentrations; the higher phytochelatin content was obtained in cultures with 4 mg Cd/L and 1 mM sulphate. The maximum EC{sub 50} value (based on nominal cadmium concentration) reached for this microalga was 4.46 ± 0.42 mg Cd/L when the sulphate concentration added to the culture medium was also 1 m

System-level network analysis of nitrogen starvation and recovery in Chlamydomonas reinhardtii reveals potential new targets for increased lipid accumulation

Czech Academy of Sciences Publication Activity Database

Valledor, Luis; Furuhashi, T.; Recuenco-Muňoz, L.; Wienkoop, S.; Weckwerth, W.

2014-01-01

Roč. 7, č. 171 (2014), s. 1-17 ISSN 1754-6834 Institutional support: RVO:67179843 Keywords : chlamydomonas reinhardtii * lipid accumulation * nitrogen Subject RIV: EI - Biotechnology ; Bionics Impact factor: 6.044, year: 2014

The small molecule fenpropimorph rapidly converts chloroplast membrane lipids to triacylglycerols in Chlamydomonas reinhardtii

Directory of Open Access Journals (Sweden)

Hanul eKim

2015-02-01

Full Text Available Concern about global warming has prompted an intense interest in developing economical methods of producing biofuels. Microalgae provide a promising platform for biofuel production, because they accumulate high levels of lipids, and do not compete with food or feed sources. However, current methods of producing algal oil involve subjecting the microalgae to stress conditions, such as nitrogen deprivation, and are prohibitively expensive. Here, we report that the fungicide fenpropimorph rapidly causes high levels of neutral lipids to accumulate in Chlamydomonas reinhardtii cells. When treated with fenpropimorph (10 μg mL–1 for 1 h, Chlamydomonas cells accumulated at least four-fold the amount of triacylglycerols (TAGs present in the untreated control cells. Furthermore, the quantity of TAGs present after 1 h of fenpropimorph treatment was over two-fold higher than that formed after 9 days of nitrogen starvation in medium with no acetate supplement. Biochemical analysis of lipids revealed that the accumulated TAGs were derived mainly from chloroplast polar membrane lipids. Such a conversion of chloroplast polar lipids to TAGs is desirable for biodiesel production, because polar lipids are usually removed during the biodiesel production process. Thus, our data exemplified that a cost and time effective method of producing TAGs is possible using fenpropimorph or similar drugs.

Dephosphorylation Pathway of D-myo-Inositol 1,4,5-trisphosphate in the Unicellular Green Alga Chlamydomonas eugametos

NARCIS (Netherlands)

Klerk, Hans; Himbergen, John A.J. van; Musgrave, Alan; Haastert, Peter J.M. van; Ende, Herman van den

In vitro dephosphorylation of D-myo-inositol 1,4,5-trisphosphate [Ins(l,4,5)P-3] by vegetative cells, gametes and zygotes of the green alga Chlamydomonas eugametos was studied using a soluble cell fraction as enzyme source and labelled Ins(1,4,5)P-3 as substrate. This compound was dephosphorylated

Chlamydomonas reinhardtii: duration of its cell cycle and phases at growth rates affected by light intensity

Czech Academy of Sciences Publication Activity Database

Vítová, Milada; Bišová, Kateřina; Umysová, Dáša; Hlavová, Monika; Kawano, S.; Zachleder, Vilém; Čížková, Mária

2011-01-01

Roč. 233, č. 1 (2011), s. 75-86 ISSN 0032-0935 R&D Projects: GA AV ČR IAA500200614; GA ČR GA525/09/0102; GA ČR GA204/09/0111 Institutional research plan: CEZ:AV0Z50200510 Keywords : Cell division timing * Cell cycle phases * Chlamydomonas Subject RIV: EE - Microbiology, Virology Impact factor: 3.000, year: 2011

The chloroplasts membrane phospholipids of Chlamydomonas reinhardii mutant not forming the Photosystem 2

International Nuclear Information System (INIS)

Trusova, V.M.; Ladygin, V.G.; Mezentsev, V.V.; Molchanov, M.I.

1987-01-01

Study on a component composition and physical state of photosynthetic membranes of Chlamydomonas chloroplasts of the wild type and mutant A-110 with disturbance of electron transfer chain in the photosystem 2 region permitted to conclude that 170 A diameter particles localized on the internal hydrophobic surface of membrane chips are deleted with respect to phosphatidylglycerin. The results obtained permit to suggest that the formation of protein-lipid complexes containing phosphatidylglycerins is suppressed in mutant A-110 which is not capable of the lamellar system differentation in

Proteomic analysis of isolated chlamydomonas centrioles reveals orthologs of ciliary-disease genes.

Science.gov (United States)

Keller, Lani C; Romijn, Edwin P; Zamora, Ivan; Yates, John R; Marshall, Wallace F

2005-06-21

The centriole is one of the most enigmatic organelles in the cell. Centrioles are cylindrical, microtubule-based barrels found in the core of the centrosome. Centrioles also act as basal bodies during interphase to nucleate the assembly of cilia and flagella. There are currently only a handful of known centriole proteins. We used mass-spectrometry-based MudPIT (multidimensional protein identification technology) to identify the protein composition of basal bodies (centrioles) isolated from the green alga Chlamydomonas reinhardtii. This analysis detected the majority of known centriole proteins, including centrin, epsilon tubulin, and the cartwheel protein BLD10p. By combining proteomic data with information about gene expression and comparative genomics, we identified 45 cross-validated centriole candidate proteins in two classes. Members of the first class of proteins (BUG1-BUG27) are encoded by genes whose expression correlates with flagellar assembly and which therefore may play a role in ciliogenesis-related functions of basal bodies. Members of the second class (POC1-POC18) are implicated by comparative-genomics and -proteomics studies to be conserved components of the centriole. We confirmed centriolar localization for the human homologs of four candidate proteins. Three of the cross-validated centriole candidate proteins are encoded by orthologs of genes (OFD1, NPHP-4, and PACRG) implicated in mammalian ciliary function and disease, suggesting that oral-facial-digital syndrome and nephronophthisis may involve a dysfunction of centrioles and/or basal bodies. By analyzing isolated Chlamydomonas basal bodies, we have been able to obtain the first reported proteomic analysis of the centriole.

Chlamydomonas IFT25 is dispensable for flagellar assembly but required to export the BBSome from flagella

Directory of Open Access Journals (Sweden)

Bin Dong

2017-11-01

Full Text Available Intraflagellar transport (IFT particles are composed of polyprotein complexes IFT-A and IFT-B as well as cargo adaptors such as the BBSome. Two IFT-B subunits, IFT25 and IFT27 were found to form a heterodimer, which is essential in exporting the BBSome out of the cilium but not involved in flagellar assembly and cytokinesis in vertebrates. Controversial results were, however, recorded to show that defects in IFT, flagellar assembly and even cytokinesis were caused by IFT27 knockdown in Chlamydomonas reinhardtii. Using C. reinhardtii as a model organism, we report that depletion of IFT25 has no effect on flagellar assembly and does not affect the entry of the BBSome into the flagellum, but IFT25 depletion did impair BBSome movement out of the flagellum, clarifying the evolutionally conserved role of IFT25 in regulating the exit of the BBSome from the flagellum cross species. Interestingly, depletion of IFT25 causes dramatic reduction of IFT27 as expected, which does not cause defects in flagellar assembly and cytokinesis in C. reinhardtii. Our data thus support that Chlamydomonas IFT27, like its vertebrate homologues, is not involved in flagellar assembly and cytokinesis.

Isolation and proteomic analysis of Chlamydomonas centrioles.

Science.gov (United States)

Keller, Lani C; Marshall, Wallace F

2008-01-01

Centrioles are barrel-shaped cytoskeletal organelles composed of nine triplet microtubules blades arranged in a pinwheel-shaped array. Centrioles are required for recruitment of pericentriolar material (PCM) during centrosome formation, and they act as basal bodies, which are necessary for the outgrowth of cilia and flagella. Despite being described over a hundred years ago, centrioles are still among the most enigmatic organelles in all of cell biology. To gain molecular insights into the function and assembly of centrioles, we sought to determine the composition of the centriole proteome. Here, we describe a method that allows for the isolation of virtually "naked" centrioles, with little to no obscuring PCM, from the green alga, Chlamydomonas. Proteomic analysis of this material provided evidence that multiple human disease gene products encode protein components of the centriole, including genes involved in Meckel syndrome and Oral-Facial-Digital syndrome. Isolated centrioles can be used in combination with a wide variety of biochemical assays in addition to being utilized as a source for proteomic analysis.

Seasonal and diel changes in photosynthetic activity of the snow algae Chlamydomonas nivalis (Chlorophyceae) from Svalbard determined by PAM fluorometry

Czech Academy of Sciences Publication Activity Database

Stibal, Marek; Elster, Josef; Šabacká, Marie; Kaštovská, Klára

2007-01-01

Roč. 59, - (2007), s. 265-273 ISSN 0168-6496 R&D Projects: GA AV ČR KJB6005409 Institutional research plan: CEZ:AV0Z60050516 Keywords : Chlamydomonas nivalis * photosynthetic activity * PAM fluorometry Subject RIV: EF - Botanics Impact factor: 3.039, year: 2007

High-yield secretion of recombinant proteins from the microalga Chlamydomonas reinhardtii

DEFF Research Database (Denmark)

Ramos Martinez, Erick Miguel; Fimognari, Lorenzo; Sakuragi, Yumiko

2017-01-01

Microalga-based biomanufacturing of recombinant proteins is attracting growing attention due to its advantages in safety, metabolic diversity, scalability and sustainability. Secretion of recombinant proteins can accelerate the use of microalgal platforms by allowing post......-translational modifications and easy recovery of products from the culture media. However, currently, the yields of secreted recombinant proteins are low, which hampers the commercial application of this strategy. This study aimed at expanding the genetic tools for enhancing secretion of recombinant proteins in Chlamydomonas...... in the endoplasmic reticulum (ER). Taken together, the results demonstrate the utility of the gametolysin signal sequence and (SP)n glycomodule to promote a more efficient biomanufacturing of microalgae-based recombinant proteins....

Productivity and selective accumulation of carotenoids of the novel extremophile microalga Chlamydomonas acidophila grown with different carbon sources in batch systems

NARCIS (Netherlands)

Cuaresma, M.; Casal, C.; Forján, E.; Vílchez, C.

2011-01-01

Cultivation of extremophile microorganisms has attracted interest due to their ability to accumulate highvalue compounds. Chlamydomonas acidophila is an acidophile green microalga isolated by our group from Tinto River, an acidic river that flows down from the mining area in Huelva, Spain. This

Dissecting the molecular mechanisms of intraflagellar transport in Chlamydomonas

DEFF Research Database (Denmark)

Pedersen, L. B.; Geimer, S.; Rosenbaum, J. L.

2006-01-01

Background The assembly and maintenance of eukaryotic cilia and flagella are mediated by intraflagellar transport (IFT), a bidirectional microtubule (MT)-based transport system. The IFT system consists of anterograde (kinesin-2) and retrograde (cDynein1b) motor complexes and IFT particles...... comprising two complexes, A and B. In the current model for IFT, kinesin-2 carries cDynein1b, IFT particles, and axonemal precursors from the flagellar base to the tip, and cDynein1b transports kinesin-2, IFT particles, and axonemal turnover products from the tip back to the base. Most of the components...... of the IFT system have been identified and characterized, but the mechanisms by which these different components are coordinated and regulated at the flagellar base and tip are unclear. Results Using a variety of Chlamydomonas mutants, we confirm that cDynein1b requires kinesin-2 for transport toward the tip...

Characterization of Hydrocortisone Biometabolites and 18S rRNA Gene in Chlamydomonas reinhardtii Cultures

Directory of Open Access Journals (Sweden)

Seyed Bagher Mosavi-Azam

2008-10-01

Full Text Available A unicellular microalga, Chlamydomonas reinhardtii, was isolated from rice paddy-field soil and water samples and used in the biotransformation of hydrocortisone (1. This strain has not been previously tested for steroid bioconversion. Fermentation was carried out in BG-11 medium supplemented with 0.05% substrate at 25ºC for 14 days of incubation. The products obtained were chromatographically purified and characterized using spectroscopic methods. 11b,17b-Dihydroxyandrost-4-en-3-one (2, 11b-hydroxyandrost-4-en-3,17-dione (3, 11b,17a,20b,21-tetrahydroxypregn-4-en-3-one (4 and prednisolone (5 were the main products of the bioconversion. The observed bioreaction features were the side chain degradation of the substrate to give compounds 2 and 3 and the 20-ketone reduction and 1,2-dehydrogenation affording compounds 4 and 5, respectively. A time course study showed the accumulation of product 2 from the second day of the fermentation and of compounds 3, 4 and 5 from the third day. All the metabolites reached their maximum concentration in seven days. Microalgal 18S rRNA gene was also amplified by PCR. PCR products were sequenced to confirm their authenticity as 18S rRNA gene of microalgae. The result of PCR blasted with other sequenced microalgae in NCBI showed 100% homology to the 18S small subunit rRNA of two Chlamydomonas reinhardtii spp.

Characterization of hydrocortisone biometabolites and 18S rRNA gene in Chlamydomonas reinhardtii cultures.

Science.gov (United States)

Ghasemi, Younes; Rasoul-Amini, Sara; Morowvat, Mohammad Hossein; Raee, Mohammad Javad; Ghoshoon, Mohammad Bagher; Nouri, Fatemeh; Negintaji, Narges; Parvizi, Rezvan; Mosavi-Azam, Seyed Bagher

2008-10-31

A unicellular microalga, Chlamydomonas reinhardtii, was isolated from rice paddy-field soil and water samples and used in the biotransformation of hydrocortisone (1). This strain has not been previously tested for steroid bioconversion. Fermentation was carried out in BG-11 medium supplemented with 0.05% substrate at 25 degrees C for 14 days of incubation. The products obtained were chromatographically purified and characterized using spectroscopic methods. 11b,17 beta-Dihydroxyandrost-4-en-3-one (2), 11 beta-hydroxyandrost-4-en-3,17-dione (3), 11 beta,17 alpha,20 beta,21-tetrahydroxypregn-4-en-3-one (4) and prednisolone (5) were the main products of the bioconversion. The observed bioreaction features were the side chain degradation of the substrate to give compounds 2 and 3 and the 20-ketone reduction and 1,2-dehydrogenation affording compounds 4 and 5, respectively. A time course study showed the accumulation of product 2 from the second day of the fermentation and of compounds 3, 4 and 5 from the third day. All the metabolites reached their maximum concentration in seven days. Microalgal 18S rRNA gene was also amplified by PCR. PCR products were sequenced to confirm their authenticity as 18S rRNA gene of microalgae. The result of PCR blasted with other sequenced microalgae in NCBI showed 100% homology to the 18S small subunit rRNA of two Chlamydomonas reinhardtii spp.

Chlamydomonas reinhardtii: the model of choice to study mitochondria from unicellular photosynthetic organisms.

Science.gov (United States)

Funes, Soledad; Franzén, Lars-Gunnar; González-Halphen, Diego

2007-01-01

Chlamydomonas reinhardtii is a model organism to study photosynthesis, cellular division, flagellar biogenesis, and, more recently, mitochondrial function. It has distinct advantages in comparison to higher plants because it is unicellular, haploid, and amenable to tetrad analysis, and its three genomes are subject to specific transformation. It also has the possibility to grow either photoautotrophically or heterotrophically on acetate, making the assembly of the photosynthetic machinery not essential for cell viability. Methods developed allow the isolation of C. reinhardtii mitochondria free of thylakoid contaminants. We review the general procedures used for the biochemical characterization of mitochondria from this green alga.

A cost-effective approach to produce 15N-labelled amino acids employing Chlamydomonas reinhardtii CC503.

Science.gov (United States)

Nicolás Carcelén, Jesús; Marchante-Gayón, Juan Manuel; González, Pablo Rodríguez; Valledor, Luis; Cañal, María Jesús; Alonso, José Ignacio García

2017-08-18

The use of enriched stable isotopes is of outstanding importance in chemical metrology as it allows the application of isotope dilution mass spectrometry (IDMS). Primary methods based on IDMS ensure the quality of the analytical measurements and traceability of the results to the international system of units. However, the synthesis of isotopically labelled molecules from enriched stable isotopes is an expensive and a difficult task. Either chemical and biochemical methods to produce labelled molecules have been proposed, but so far, few cost-effective methods have been described. The aim of this study was to use the microalgae Chlamydomonas reinhardtii to produce, at laboratory scale, 15 N-labelled amino acids with a high isotopic enrichment. To do that, a culture media containing 15 NH 4 Cl was used. No kinetic isotope effect (KIE) was observed. The labelled proteins biosynthesized by the microorganism were extracted from the biomass and the 15 N-labelled amino acids were obtained after a protein hydrolysis with HCl. The use of the wall deficient strain CC503 cw92 mt+ is fit for purpose, as it only assimilates ammonia as nitrogen source, avoiding isotope contamination with nitrogen from the atmosphere or the reagents used in the culture medium, and enhancing the protein extraction efficiency compared to cell-walled wild type Chlamydomonas. The isotopic enrichment of the labelled amino acids was calculated from their isotopic composition measured by gas chromatography mass spectrometry (GC-MS). The average isotopic enrichment for the 16 amino acids characterized was 99.56 ± 0.05% and the concentration of the amino acids in the hydrolysate ranged from 18 to 90 µg/mL. Previously reported biochemical methods to produce isotopically labelled proteins have been applied in the fields of proteomics and fluxomics. For these approaches, low amounts of products are required and the isotopic enrichment of the molecules has never been properly determined. So far, only 13

Effect of mutagen combined action on Chlamydomonas Reinhardtii cells. I

International Nuclear Information System (INIS)

Vlcek, D.; Podstavkova, S.; Dubovsky, J.

1978-01-01

The effect was investigated of single and combined actions of alkylnitrosourea derivatives (N-methyl-N-nitrosourea and N-ethyl-N-nitrosourea) and UV-radiation on the survival of cells of Chlamydomonas reinhardtii algae in dependence on the sequence of application of mutagens and on the given conditions of cultivation following mutagen activity. In particular, the single phases were investigated of the total lethal effect, i.e., the death of cells before division and their death after division. The most pronounced changes in dependence on the sequence of application of mutagens and on the given conditions of cultivation were noted in cell death before division. In dependence on the sequence of application of mutagens, the effect of the combined action on the survival of cells changed from an additive (alkylnitrosourea + UV-radiation) to a protective effect (UV-radiation + alkylnitrosourea). (author)

The Chlamydomonas cell wall and its constituent glycoproteins analyzed by the quick-freeze, deep-etch technique

OpenAIRE

1985-01-01

Using the quick-freeze, deep-etch technique, we have analyzed the structure of the intact cell wall of Chlamydomonas reinhardi, and have visualized its component glycoproteins after mechanical shearing and after depolymerization induced by perchlorate or by the wall-disrupting agent, autolysin. The intact wall has previously been shown in a thin- section study (Roberts, K., M. Gurney-Smith, and G. J. Hills, 1972, J. Ultrastruct. Res. 40:599-613) to consist of a discrete central triplet bisect...

Deletion of CGLD1 Impairs PSII and Increases Singlet Oxygen Tolerance of Green Alga Chlamydomonas reinhardtii

Directory of Open Access Journals (Sweden)

Jiale Xing

2017-12-01

Full Text Available The green alga Chlamydomonas reinhardtii is a key model organism for studying photosynthesis and oxidative stress in unicellular eukaryotes. Using a forward genetics approach, we have identified and characterized a mutant x32, which lacks a predicted protein named CGLD1 (Conserved in Green Lineage and Diatom 1 in GreenCut2, under normal and stress conditions. We show that loss of CGLD1 resulted in minimal photoautotrophic growth and PSII activity in the organism. We observed reduced amount of PSII complex and core subunits in the x32 mutant based on blue-native (BN/PAGE and immunoblot analysis. Moreover, x32 exhibited increased sensitivity to high-light stress and altered tolerance to different reactive oxygenic species (ROS stress treatments, i.e., decreased resistance to H2O2/or tert-Butyl hydroperoxide (t-BOOH and increased tolerance to neutral red (NR and rose bengal (RB that induce the formation of singlet oxygen, respectively. Further analysis via quantitative real-time PCR (qRT-PCR indicated that the increased singlet-oxygen tolerance of x32 was largely correlated with up-regulated gene expression of glutathione-S-transferases (GST. The phenotypical and physiological implications revealed from our experiments highlight the important roles of CGLD1 in maintaining structure and function of PSII as well as in protection of Chlamydomonas under photo-oxidative stress conditions.

Development of phytase-expressing chlamydomonas reinhardtii for monogastric animal nutrition.

Science.gov (United States)

Erpel, Fernanda; Restovic, Franko; Arce-Johnson, Patricio

2016-03-12

In plant-derived animal feedstuffs, nearly 80 % of the total phosphorus content is stored as phytate. However, phytate is poorly digested by monogastric animals such as poultry, swine and fish, as they lack the hydrolytic enzyme phytase; hence it is regarded as a nutritionally inactive compound from a phosphate bioavailability point of view. In addition, it also chelates important dietary minerals and essential amino acids. Therefore, dietary supplementation with bioavailable phosphate and exogenous phytases are required to achieve optimal animal growth. In order to simplify the obtaining and application processes, we developed a phytase expressing cell-wall deficient Chlamydomonas reinhardtii strain. In this work, we developed a transgenic microalgae expressing a fungal phytase to be used as a food supplement for monogastric animals. A codon optimized Aspergillus niger PhyA E228K phytase (mE228K) with improved performance at pH 3.5 was transformed into the plastid genome of Chlamydomonas reinhardtii in order to achieve optimal expression. We engineered a plastid-specific construction harboring the mE228K gene, which allowed us to obtain high expression level lines with measurable in vitro phytase activity. Both wild-type and cell-wall deficient strains were selected, as the latter is a suitable model for animal digestion. The enzymatic activity of the mE228K expressing lines were approximately 5 phytase units per gram of dry biomass at pH 3.5 and 37 °C, similar to physiological conditions and economically competitive for use in commercial activities. A reference basis for the future biotechnological application of microalgae is provided in this work. A cell-wall deficient transgenic microalgae with phytase activity at gastrointestinal pH and temperature and suitable for pellet formation was developed. Moreover, the associated microalgae biomass costs of this strain would be between US$5 and US$60 per ton of feedstuff, similar to the US$2 per ton of feedstuffs

Effect of mutagen combined action on Chlamydomonas reinhardtii cells. II

International Nuclear Information System (INIS)

Podstavkova, S.; Vlcek, D.; Dubovsky, J.

1978-01-01

The effect of UV radiation and UV radiation combined with alkylnitrosourea derivatives (N-methyl-N-nitrosourea and N-ethyl-N-nitrosourea) was observed on survival of cells of the algae Chlamydomonas reinhardtii. In particular, single parts were evaluated of the overall lethal effect - dying of cells before division and dying of cells after division. It was found that the combined action of low doses of UV radiation and alkylnitrosoureas result in a pronounced protective effect which manifests itself by a higher frequency of surviving cells than was that effected by the action of alkylnitrosoureas alone. As a result of combined action with higher doses of UV radiation this effect is lost, and the resultant values will come close to the theoretically anticipated values. This gradual transition from a protective to an additive effect mainly manifests itself by changes in the proportion of cells dying before division. (author)

Systems Biology of Lipid Body Formation in the Green Alga Chlamydomonas reinhardtii

Energy Technology Data Exchange (ETDEWEB)

Goodenough, Ursula [Washington Univ., St. Louis, MO (United States)

2017-11-10

The project aimed to deepen our understanding of alga triacylglycerol (TAG) production to undergird explorations of using algal TAG as a source of biodiesel fuel. Our published contributions included the following: 1) Development of a rapid assay for TAG in algal cultures which was widely distributed to the algal community. 2) A comprehensive transcriptome analysis of the development of the ultra-high-TAG “obese” phenotype In Chlamydomonas reinhardtii. 3) A comprehensive biochemical and ultrastructural analysis of the cell wall of Nannochloropsis gaditana, whose walls render it both growth-hardy and difficult to rupture for TAG recovery. A manuscript in preparation considers the autophagy response in C. reinhardtii and its entrance into stationary phase, both having an impact on TAG production.

A revised mineral nutrient supplement increases biomass and growth rate in Chlamydomonas reinhardtii.

Science.gov (United States)

Kropat, Janette; Hong-Hermesdorf, Anne; Casero, David; Ent, Petr; Castruita, Madeli; Pellegrini, Matteo; Merchant, Sabeeha S; Malasarn, Davin

2011-06-01

Interest in exploiting algae as a biofuel source and the role of inorganic nutrient deficiency in inducing triacylglyceride (TAG) accumulation in cells necessitates a strategy to efficiently formulate species-specific culture media that can easily be manipulated. Using the reference organism Chlamydomonas reinhardtii, we tested the hypothesis that modeling trace element supplements after the cellular ionome would result in optimized cell growth. We determined the trace metal content of several commonly used Chlamydomonas strains in various culture conditions and developed a revised trace element solution to parallel these measurements. Comparison of cells growing in the revised supplement versus a traditional trace element solution revealed faster growth rates and higher maximum cell densities with the revised recipe. RNA-seq analysis of cultures growing in the traditional versus revised medium suggest that the variation in transcriptomes was smaller than that found between different wild-type strains grown in traditional Hutner's supplement. Visual observation did not reveal defects in cell motility or mating efficiency in the new supplement. Ni²⁺-inducible expression from the CYC6 promoter remained a useful tool, albeit with an increased requirement for Ni²⁺ because of the introduction of an EDTA buffer system in the revised medium. Other advantages include more facile preparation of trace element stock solutions, a reduction in total chemical use, a more consistent batch-to-batch formulation and long-term stability (tested up to 5 years). Under the new growth regime, we analyzed cells growing under different macro- and micronutrient deficiencies. TAG accumulation in N deficiency is comparable in the new medium. Fe and Zn deficiency also induced TAG accumulation, as suggested by Nile Red staining. This approach can be used to efficiently optimize culture conditions for other algal species to improve growth and to assay cell physiology. © 2011 The Authors

A Chlamydomonas-derived Human Papillomavirus 16 E7 vaccine induces specific tumor protection.

Directory of Open Access Journals (Sweden)

Olivia C Demurtas

Full Text Available The E7 protein of the Human Papillomavirus (HPV type 16, being involved in malignant cellular transformation, represents a key antigen for developing therapeutic vaccines against HPV-related lesions and cancers. Recombinant production of this vaccine antigen in an active form and in compliance with good manufacturing practices (GMP plays a crucial role for developing effective vaccines. E7-based therapeutic vaccines produced in plants have been shown to be active in tumor regression and protection in pre-clinical models. However, some drawbacks of in whole-plant vaccine production encouraged us to explore the production of the E7-based therapeutic vaccine in Chlamydomonas reinhardtii, an organism easy to grow and transform and fully amenable to GMP guidelines.An expression cassette encoding E7GGG, a mutated, attenuated form of the E7 oncoprotein, alone or as a fusion with affinity tags (His6 or FLAG, under the control of the C. reinhardtii chloroplast psbD 5' UTR and the psbA 3' UTR, was introduced into the C. reinhardtii chloroplast genome by homologous recombination. The protein was mostly soluble and reached 0.12% of total soluble proteins. Affinity purification was optimized and performed for both tagged forms. Induction of specific anti-E7 IgGs and E7-specific T-cell proliferation were detected in C57BL/6 mice vaccinated with total Chlamydomonas extract and with affinity-purified protein. High levels of tumor protection were achieved after challenge with a tumor cell line expressing the E7 protein.The C. reinhardtii chloroplast is a suitable expression system for the production of the E7GGG protein, in a soluble, immunogenic form. The production in contained and sterile conditions highlights the potential of microalgae as alternative platforms for the production of vaccines for human uses.

Using single cell cultivation system for on-chip monitoring of the interdivision timer in Chlamydomonas reinhardtii cell cycle

Directory of Open Access Journals (Sweden)

Soloviev Mikhail

2010-09-01

Full Text Available Abstract Regulation of cell cycle progression in changing environments is vital for cell survival and maintenance, and different regulation mechanisms based on cell size and cell cycle time have been proposed. To determine the mechanism of cell cycle regulation in the unicellular green algae Chlamydomonas reinhardtii, we developed an on-chip single-cell cultivation system that allows for the strict control of the extracellular environment. We divided the Chlamydomonas cell cycle into interdivision and division phases on the basis of changes in cell size and found that, regardless of the amount of photosynthetically active radiation (PAR and the extent of illumination, the length of the interdivision phase was inversely proportional to the rate of increase of cell volume. Their product remains constant indicating the existence of an 'interdivision timer'. The length of the division phase, in contrast, remained nearly constant. Cells cultivated under light-dark-light conditions did not divide unless they had grown to twice their initial volume during the first light period. This indicates the existence of a 'commitment sizer'. The ratio of the cell volume at the beginning of the division phase to the initial cell volume determined the number of daughter cells, indicating the existence of a 'mitotic sizer'.

Effect of red and blue light on the timing of cyclin-dependent kinase activity and the timing of cell division in Chlamydomonas reinhardtii

Czech Academy of Sciences Publication Activity Database

Oldenhof, H.; Bišová, Kateřina; Ende, H.; Zachleder, Vilém

2004-01-01

Roč. 42, - (2004), s. 341-348 ISSN 0981-9428 R&D Projects: GA ČR GA204/02/1438 Institutional research plan: CEZ:AV0Z5020903 Keywords : blue light * chlamydomonas reingardtii * cell cycle Subject RIV: EA - Cell Biology Impact factor: 1.414, year: 2004

Flocculation of Chlamydomonas reinhardtii with Different Phenotypic Traits by Metal Cations and High pH

Directory of Open Access Journals (Sweden)

Jianhua Fan

2017-11-01

Full Text Available Concentrating algal cells by flocculation as a prelude to centrifugation could significantly reduce the energy and cost of harvesting the algae. However, how variation in phenotypic traits such as cell surface features, cell size and motility alter the efficiency of metal cation and pH-induced flocculation is not well understood. Our results demonstrate that both wild-type and cell wall-deficient strains of the green unicellular alga Chlamydomonas reinhardtii efficiently flocculate (>90% at an elevated pH of the medium (pH 11 upon the addition of divalent cations such as calcium and magnesium (>5 mM. The trivalent ferric cation (at 10 mM proved to be essential for promoting flocculation under weak alkaline conditions (pH ∼8.5, with a maximum efficiency that exceeded 95 and 85% for wild-type CC1690 and the cell wall-deficient sta6 mutant, respectively. Near complete flocculation could be achieved using a combination of 5 mM calcium and a pH >11, while the medium recovered following cell removal could be re-cycled without affecting algal growth rates. Moreover, the absence of starch in the cell had little overall impact on flocculation efficiency. These findings contribute to our understanding of flocculation in different Chlamydomonas strains and have implications with respect to inexpensive methods for harvesting algae with different phenotypic traits. Additional research on the conditions (e.g., pH and metal ions used for efficient flocculation of diverse algal groups with diverse characteristics, at both small and large scale, will help establish inexpensive procedures for harvesting cell biomass.

Effect of temperature and light intensity on growth and photosynthetic activity of Chlamydomonas Reinhardtii

International Nuclear Information System (INIS)

Alfonsel, M.; Fernandez Gonzalez, J.

1986-01-01

The effect of five temperatures (15, 20, 25, 30 and 35 0 C) and two levels of illumination on growth and photosynthetic activity of Chlamydomonas reinhardtii has been studied. The growth of the cultures was evaluated by optical density. Photosynthetic activity has been carried out studying either the assimilation rate of CO 2 labelled with C 14 or the oxygen evolution by means of polarographic measurements. The maximum photosynthetic rate has been obtained at 25 0 C for the lower lavel of illumination (2400 lux) and at 35 0 C for the higher one (13200 lux). These results suggest an interacton of temperature and illumination on photosynthetic activity. (author)

Uptake pf 203Hg++ and sup(115M)Cd++ by growing chlamydomonas reinhardi under different conditions

International Nuclear Information System (INIS)

Macka, W.; Stehlik, G.; Whilidal, H.; Washuettl, J.; Bancher, E.

1977-09-01

The uptake of labelled mercury and cadmium ions by living and dead cells Chlamydomonas reinhardi WT + was measured at 25 deg C in minimal and optimal conditions (dark/nitrogen or light/air, respectively). In each case incorporation was completely independent of external energy. Living and dead cells incorporate almost the same amount of the added heavy metal ions; after about 4 to 8 hours saturation was obtained. Furthermore the distribution of mercury in the system alga/culture medium/gas phase and the amount of mercury and cadmium adsorbed by cells were studied. (author)

Chlamydomonas DYX1C1/PF23 is essential for axonemal assembly and proper morphology of inner dynein arms.

Directory of Open Access Journals (Sweden)

Ryosuke Yamamoto

2017-09-01

Full Text Available Cytoplasmic assembly of ciliary dyneins, a process known as preassembly, requires numerous non-dynein proteins, but the identities and functions of these proteins are not fully elucidated. Here, we show that the classical Chlamydomonas motility mutant pf23 is defective in the Chlamydomonas homolog of DYX1C1. The pf23 mutant has a 494 bp deletion in the DYX1C1 gene and expresses a shorter DYX1C1 protein in the cytoplasm. Structural analyses, using cryo-ET, reveal that pf23 axonemes lack most of the inner dynein arms. Spectral counting confirms that DYX1C1 is essential for the assembly of the majority of ciliary inner dynein arms (IDA as well as a fraction of the outer dynein arms (ODA. A C-terminal truncation of DYX1C1 shows a reduction in a subset of these ciliary IDAs. Sucrose gradients of cytoplasmic extracts show that preassembled ciliary dyneins are reduced compared to wild-type, which suggests an important role in dynein complex stability. The role of PF23/DYX1C1 remains unknown, but we suggest that DYX1C1 could provide a scaffold for macromolecular assembly.

Physiological and Biochemical characterization of Chlamydomonas sp. the Hydrogen Production's Strain

International Nuclear Information System (INIS)

Chader, S.; Belhamel, M.; H Hacene

2006-01-01

The hydrogen produced by biological way became, one of the most interesting subjects of research relating to development the energy system starting from renewable sources. This study describes the closed relation between the physiological behaviour, biochemical and rate of gases produced by Chlamydomonas sp. strain AT14, isolated in the area of Touat (the Sahara Algerian) and cultivated in a toric photo-bioreactor. A considerable growth was noted, where the concentration of the biomass double in only two days after incubation. The micro-algal cells present a 100% of viability, which relocate has satisfactory behaviour in the toric engine. In addition, the displacement water level in the system of measurement implies has gas production (0.1 ml) in coordination with the anaerobic period of the reactional enclosure. The yield of this way of hydrogen production is depending on the species used, the light intensity, and the conditions of culture. (authors)

Cell growth and protein synthesis of unicellular green alga Chlamydomonas in heavy water

International Nuclear Information System (INIS)

Ishida, M.R.

1983-01-01

The effects of heavy water on the cell growth and protein synthesis of the photoautotrophically growing Chlamydomonas cells were studied. The growth rate of the cells is inversely proportional to the concentrations of heavy water. The cells can barely live in 90% heavy water, but they die in 99.85% heavy water within a few days. Incorporation of 14 Cleucine into cells is markedly stimulated by heavy water of various concentrations between 30 and 99.85% in the case of the short time incubation. Contrary to this, in the long time incubation as several days, heavy water inhibits the protein synthesis. Such two modes of the protein synthetic activities are dependent upon the incubation time of the cells grown photoautotrophically in the heavy water media. (author)

Taxonomic identity and physiological ecology of Chlamydomonas hedleyi sp. nov. , algal flagellate symbiont from the foraminifer Archaias angulatus

Energy Technology Data Exchange (ETDEWEB)

Lee, J J; Crockett, L J; Hagen, J; Stone, R J

1975-12-31

The fine structure of the symbiotic alga isolated from the foraminiferan Archaias angulatus (Fichtel et Moll) DeMontfort is typical of the Chlorophyceae of the volvocalean and chlorococcalean lines. Spherical non-motile cells, 10--14 ..mu..m in diameter, characterize the dominant life cycle phase. Long oval motile forms with truncated apices are present 3--5 days after transfer to fresh medium. The pyrenoids are embedded anteriorly in the singly bilobed chloroplast and are surrounded by a sheath of starch platelets. In spite of the non-motile state of cells in older cultures (which is perhaps a reflection of its normally symbiotic condition), the alga is identified as a species of the volvocalean genus Chlamydomonas and is named C. hedleyi sp. nov. The symbiont has no vitamin or organic requirements but growth is increased threefold in the presence of thiamine, and twofold in the presence of 1 ..mu..m glutamic acid, histidine and methionine. Urea was the best nitrogen source tested. Purines and pyrimidines did not serve as nitrogen sources. Chlamydomonas hedleyi grows well in a salinity range of 6- greater than 52 per thousand and a pH range of 6--8.5. 7.04 x 10/sup -7/ M carbon h/sup -1/ g/sup -1/ was fixed by the symbiont, 57 percent being released into the medium as a chromatographically homogeneous organic molecule provisionally identified as mannitol.

Response of Chlamydomonas reinhardtii to naphthenic acid exposure

Energy Technology Data Exchange (ETDEWEB)

Goff, K.; Wilson, K. [Saskatchewan Univ., Saskatoon, SK (Canada); Headley, J. [Environment Canada, Ottawa, ON (Canada)

2010-07-01

This study examined the feasibility of using a model organism for the algal bioremediation of oil sands process water (OSPW), a highly toxic mixture of sediments, bitumen, ions, and organic and inorganic compounds. Naphthenic acids (NAs) are a contaminant class of particular concern. Bioremediation techniques may mitigate toxicity of OSPW in general, and NAs in particular. Although most studies on the biodegradation of NAs focus on the role of bacteria, fungi, and emergent macrophytes, studies have indicated that algae may also play a key role through direct degradation, biosequestration, or photosynthetic aeration of waters to promote other biological reactions. Chlamydomonas frigida is of particular interest, but no cultures are currently available. Therefore, this study used C. reinhardtii, a well-characterized model organism, to begin analysis of potential algal bioremediation of OSPW. Cultures of C. reinhardtii were grown heterotrophically in nutrient media spiked with a dilution series of NAs. Culture densities were measured to compile growth curves over time, changes in rate of growth, and survivability. Negative ion electrospray mass spectrometry was used to determine the concentration of NAs in solution in relation to growth rate and culture density. The study determined the tolerance of C. reinhardtii to NAs. A mechanism for this tolerance was then proposed.

Histones of Chlamydomonas reinhardtii. Synthesis, acetylation, and methylation

International Nuclear Information System (INIS)

Waterborg, J.H.; Robertson, A.J.; Tatar, D.L.; Borza, C.M.; Davie, J.R.

1995-01-01

Histones of the green alga Chlamydomonas reinhardtii were prepared by a new method and fractionated by reversed-phase high-performance liquid chromatography. Acid-urea-Triton gel analysis and tritiated acetate labeling demonstrated high levels of steady-state acetylation for the single histone H3 protein, in contrast to low levels on histones H4 and H2B. Twenty percent of histone H3 is subject to dynamic acetylation with, on average, three acetylated lysine residues per protein molecule. Histone synthesis in light-dark-synchronized cultures was biphasic with pattern differences between two histone H1 variants, between two H2A variants, and between H2B and ubiquitinated H2B. Automated protein sequence analysis of histone H3 demonstrated a site-specific pattern of steady-state acetylation between 7 and 17% at five of the six amino-terminal lysines and of monomethylation between 5 and 81% at five of the eight amino-terminal lysines in a pattern that may limit dynamic acetylation. An algal histone H3 sequence was confirmed by protein sequencing with a since threonine as residue 28 instead of the serine(28)-alanine(29) sequence, present in all other known plant and animal H3 histones

Cellulose degradation and assimilation by the unicellular phototrophic eukaryote Chlamydomonas reinhardtii.

Science.gov (United States)

Blifernez-Klassen, Olga; Klassen, Viktor; Doebbe, Anja; Kersting, Klaudia; Grimm, Philipp; Wobbe, Lutz; Kruse, Olaf

2012-01-01

Plants convert sunlight to biomass, which is primarily composed of lignocellulose, the most abundant natural biopolymer and a potential feedstock for fuel and chemical production. Cellulose assimilation has so far only been described for heterotrophic organisms that rely on photosynthetically active primary producers of organic compounds. Among phototrophs, the unicellular green microalga Chlamydomonas reinhardtii is widely known as one of the best established model organisms. It occupies many habitats, including aquatic and soil ecosystems. This ubiquity underscores the versatile metabolic properties of this microorganism. Here we present yet another paradigm of adaptation for C. reinhardtii, highlighting its photoheterotrophic ability to utilize cellulose for growth in the absence of other carbon sources. When grown under CO(2)-limiting conditions in the light, secretion of endo-β-1,4-glucanases by the cell causes digestion of exogenous cellulose, followed by cellobiose uptake and assimilation. Phototrophic microbes like C. reinhardtii may thus serve as biocatalysts for cellulosic biofuel production.

Lipidomic Analysis of Chlamydomonas reinhardtii under Nitrogen and Sulfur Deprivation.

Directory of Open Access Journals (Sweden)

Dawei Yang

Full Text Available Chlamydomonas reinhardtii accumulates lipids under complete nutrient starvation conditions while overall growth in biomass stops. In order to better understand biochemical changes under nutrient deprivation that maintain production of algal biomass, we used a lipidomic assay for analyzing the temporal regulation of the composition of complex lipids in C. reinhardtii in response to nitrogen and sulfur deprivation. Using a chip-based nanoelectrospray direct infusion into an ion trap mass spectrometer, we measured a diversity of lipid species reported for C. reinhardtii, including PG phosphatidylglycerols, PI Phosphatidylinositols, MGDG monogalactosyldiacylglycerols, DGDG digalactosyldiacylglycerols, SQDG sulfoquinovosyldiacylglycerols, DGTS homoserine ether lipids and TAG triacylglycerols. Individual lipid species were annotated by matching mass precursors and MS/MS fragmentations to the in-house LipidBlast mass spectral database and MS2Analyzer. Multivariate statistics showed a clear impact on overall lipidomic phenotypes on both the temporal and the nutrition stress level. Homoserine-lipids were found up-regulated at late growth time points and higher cell density, while triacyclglycerols showed opposite regulation of unsaturated and saturated fatty acyl chains under nutritional deprivation.

The mechanism of photosystem-II inactivation during sulphur deprivation-induced H2 production in Chlamydomonas reinhardtii.

Science.gov (United States)

Nagy, Valéria; Vidal-Meireles, André; Podmaniczki, Anna; Szentmihályi, Klára; Rákhely, Gábor; Zsigmond, Laura; Kovács, László; Tóth, Szilvia Z

2018-05-01

Sulphur limitation may restrain cell growth and viability. In the green alga Chlamydomonas reinhardtii, sulphur limitation may induce H 2 production lasting for several days, which can be exploited as a renewable energy source. Sulphur limitation causes a large number of physiological changes, including the inactivation of photosystem II (PSII), leading to the establishment of hypoxia, essential for the increase in hydrogenase expression and activity. The inactivation of PSII has long been assumed to be caused by the sulphur-limited turnover of its reaction center protein PsbA. Here we reinvestigated this issue in detail and show that: (i) upon transferring Chlamydomonas cells to sulphur-free media, the cellular sulphur content decreases only by about 25%; (ii) as demonstrated by lincomycin treatments, PsbA has a significant turnover, and other photosynthetic subunits, namely RbcL and CP43, are degraded more rapidly than PsbA. On the other hand, sulphur limitation imposes oxidative stress early on, most probably involving the formation of singlet oxygen in PSII, which leads to an increase in the expression of GDP-L-galactose phosphorylase, playing an essential role in ascorbate biosynthesis. When accumulated to the millimolar concentration range, ascorbate may inactivate the oxygen-evolving complex and provide electrons to PSII, albeit at a low rate. In the absence of a functional donor side and sufficient electron transport, PSII reaction centers are inactivated and degraded. We therefore demonstrate that the inactivation of PSII is a complex and multistep process, which may serve to mitigate the damaging effects of sulphur limitation. © 2018 The Authors The Plant Journal © 2018 John Wiley & Sons Ltd.

Effect of Temperature and light intensity on growth and Photosynthetic Activity of Chlamydomonas reinhard II; Efecto de la temperatura e intensidad luminosa sobre el crecimiento y actividad fotosintetica del alga Chlamydomonas Reinhardt II

Energy Technology Data Exchange (ETDEWEB)

Alfonsel Jaen, M; Fernandez Gonzalez, J

1985-07-01

The effect of five temperatures (15,20,25,30 and 35 degree centigree) and two levels of illumination on growth and photosynthetic activity of Chlamydomonas reinhard II has been studied. The growth of the cultures was evaluated by optical density. Photosynthetic activity has been carried out studying either the assimilation rate of C0{sub 2} labelled with C-14 or the oxygen evolution by means of polarographic measurements. The maximum photosynthetic rate has been obtained at 25 degree centigree for the lower level of illumination (2400 lux) and at 35 degree centigree for the higher one (13200 lux) and at 35 degree centigree for the higher ono (13200 lux). These results suggest an interaction of temperature and illumination on photosynthetic activity. (Author) 37 refs.

Metabolic acclimation to excess light intensity in Chlamydomonas reinhardtii.

Science.gov (United States)

Davis, Maria C; Fiehn, Oliver; Durnford, Dion G

2013-07-01

There are several well-described acclimation responses to excess light in green algae but the effect on metabolism has not been thoroughly investigated. This study examines the metabolic changes during photoacclimation to high-light (HL) stress in Chlamydomonas reinhardtii using nuclear magnetic resonance and mass spectrometry. Using principal component analysis, a clear metabolic response to HL intensity was observed on global metabolite pools, with major changes in the levels of amino acids and related nitrogen metabolites. Amino acid pools increased during short-term photoacclimation, but were especially prominent in HL-acclimated cultures. Unexpectedly, we observed an increase in mitochondrial metabolism through downstream photorespiratory pathways. The expression of two genes encoding key enzymes in the photorespiratory pathway, glycolate dehydrogenase and malate synthase, were highly responsive to the HL stress. We propose that this pathway contributes to metabolite pools involved in nitrogen assimilation and may play a direct role in photoacclimation. Our results suggest that primary and secondary metabolism is highly pliable and plays a critical role in coping with the energetic imbalance during HL exposure and a necessary adjustment to support an increased growth rate that is an effective energy sink for the excess reducing power generated during HL stress. © 2013 John Wiley & Sons Ltd.

Radiocesium bioaccumulation in freshwater plankton: Influences of cation concentrations (K+ and Na+) on direct uptake of 137Cs in Chlamydomonas, Scenedesmus and Daphnia. Food-chain transfer of 137Cs from Chlamydomonas to Daphnia at different K+ concentrations

International Nuclear Information System (INIS)

Hagstroem, J.

2002-01-01

The influences of cation concentrations (K + and Na + ) on radiocesium ( 137 Cs) bioaccumulation in two freshwater phytoplankton species (Scenedesmus quadricauda and Chlamydomonas noctigama) were systematically investigated in batch-cultures monitored during two weeks. Both species were cultured at 9 μE M -2 s -1 constant illumination at 20 deg. C. The exponential growth phase lasted for more than 100 hours (μ ≅ 0.02 h -1 for C. noctigama and 0.03 h -1 for S, quadricauda). Over cation concentration ranges encountered in natural fresh waters ([K + ] from 0.1 μM to 3 mM, [Na + ] from 20 μM to 3 mM), a more than three order of magnitude variation was found for both intake rate and observed bioconcentration factors (BCF) at apparent steady-state (from less than 10 3 to 10 6 L (kg C) -1 ). For both species, the major effector on BCF and uptake rate was external [K + ], which was inversely proportional to these parameters over wide ranges (1-1000 μM for S. quadricauda and 0.1 to 300 μM for C. noctigama). At concentrations above these ranges K + still reduced 137 Cs bio-uptake, but less effectively. A minor influence of external [Na + ] on 137 Cs bioaccumulation was indicated for S. quadricauda, whereas no such influence was significant for C. noctigama. A biphasic pattern for 137 Cs bioaccumulation was discovered in C. noctigama. A rapid 'quasi-steady state' with an effective equilibration time of less than 100 hours was approached during the exponential growth phase. A surge in the uptake occurred when exponential growth ceased, and this pattern was consistent over the range 30 μM to 1.4 mM external [K + ]. Since depletion of external [K + ] was not detected for these treatments, this pattern can only be explained if there are at least two different cellular compartments involved. Although less certain, a second steady-state BCF appeared within two weeks, which seems to be up to one order of magnitude higher than the first. Microcosm experiments with the

Crystallization and preliminary X-ray characterization of full-length Chlamydomonas reinhardtii centrin

International Nuclear Information System (INIS)

Alfaro, Elisa; Valle Sosa, Liliana del; Sanoguet, Zuleika; Pastrana-Ríos, Belinda; Schreiter, Eric R.

2008-01-01

C. reinhardtii centrin, an EF-hand calcium-binding protein localized to the microtubule-organizing center of eukaryotic organisms, has been crystallized in the presence of the model peptide melittin. X-ray diffraction data were collected to 2.2 Å resolution. Chlamydomonas reinhardtii centrin is a member of the EF-hand calcium-binding superfamily. It is found in the basal body complex and is important for flagellar motility. Like other members of the EF-hand family, centrin interacts with and modulates the function of other proteins in a calcium-dependent manner. To understand how C. reinhardtii centrin interacts with its protein targets, it has been crystallized in the presence of the model peptide melittin and X-ray diffraction data have been collected to 2.2 Å resolution. The crystals are orthorhombic, with unit-cell parameters a = 52.1, b = 114.4, c = 34.8 Å, and are likely to belong to space group P2 1 2 1 2

Inhomogeneous distribution of Chlamydomonas in a cylindrical container with a bubble plume

Science.gov (United States)

Nonaka, Yuki; Kikuchi, Kenji; Numayama-Tsuruta, Keiko; Kage, Azusa; Ueno, Hironori; Ishikawa, Takuji

2016-01-01

ABSTRACT Swimming microalgae show various taxes, such as phototaxis and gravitaxis, which sometimes result in the formation of a cell-rich layer or a patch in a suspension. Despite intensive studies on the effects of shear flow and turbulence on the inhomogeneous distribution of microalgae, the effect of a bubble plume has remained unclear. In this study, we used Chlamydomonas as model microalgae, and investigated the spatial distribution of cells in a cylindrical container with a bubble plume. The results illustrate that cells become inhomogeneously distributed in the suspension due to their motility and photo-responses. A vortical ring distribution was observed below the free surface when the bubble flow rate was sufficiently small. We performed a scaling analysis on the length scale of the vortical ring, which captured the main features of the experimental results. These findings are important in understanding transport phenomena in a microalgae suspension with a bubble plume. PMID:26787679

Separation Options for Phosphorylated Osteopontin from Transgenic Microalgae Chlamydomonas reinhardtii

Directory of Open Access Journals (Sweden)

Ayswarya Ravi

2018-02-01

Full Text Available Correct folding and post-translational modifications are vital for therapeutic proteins to elicit their biological functions. Osteopontin (OPN, a bone regenerative protein present in a range of mammalian cells, is an acidic phosphoprotein with multiple potential phosphorylation sites. In this study, the ability of unicellular microalgae, Chlamydomonas reinhardtii, to produce phosphorylated recombinant OPN in its chloroplast is investigated. This study further explores the impact of phosphorylation and expression from a “plant-like” algae on separation of OPN. Chromatography resins ceramic hydroxyapatite (CHT and Gallium-immobilized metal affinity chromatography (Ga-IMAC were assessed for their binding specificity to phosphoproteins. Non-phosphorylated recombinant OPN expressed in E. coli was used to compare the specificity of interaction of the resins to phosphorylated OPN. We observed that CHT binds OPN by multimodal interactions and was better able to distinguish phosphorylated proteins in the presence of 250 mM NaCl. Ga-IMAC interaction with OPN was not selective to phosphorylation, irrespective of salt, as the resin bound OPN from both algal and bacterial sources. Anion exchange chromatography proved an efficient capture method to partially separate major phosphorylated host cell protein impurities such as Rubisco from OPN.

Chlamydomonas as a model for biofuels and bio-products production.

Science.gov (United States)

Scranton, Melissa A; Ostrand, Joseph T; Fields, Francis J; Mayfield, Stephen P

2015-05-01

Developing renewable energy sources is critical to maintaining the economic growth of the planet while protecting the environment. First generation biofuels focused on food crops like corn and sugarcane for ethanol production, and soybean and palm for biodiesel production. Second generation biofuels based on cellulosic ethanol produced from terrestrial plants, has received extensive funding and recently pilot facilities have been commissioned, but to date output of fuels from these sources has fallen well short of what is needed. Recent research and pilot demonstrations have highlighted the potential of algae as one of the most promising sources of sustainable liquid transportation fuels. Algae have also been established as unique biofactories for industrial, therapeutic, and nutraceutical co-products. Chlamydomonas reinhardtii's long established role in the field of basic research in green algae has paved the way for understanding algal metabolism and developing genetic engineering protocols. These tools are now being utilized in C. reinhardtii and in other algal species for the development of strains to maximize biofuels and bio-products yields from the lab to the field. © 2015 The Authors The Plant Journal © 2015 John Wiley & Sons Ltd.

EFFECT OF TREATED DOMESTIC WASTEWATER USED AS CULTURE MEDIUM ON THE GROWTH AND PRODUCTIVITY OF Chlamydomonas sp. STRAIN ISOLATED FROM LANDFILL LEACHATE

Directory of Open Access Journals (Sweden)

Fábio de Farias Neves

2013-07-01

Full Text Available Microalgae have been culturing to fix carbon and produce biofuels from the biomass. However, it is important to develop low cost strategies for microalgae production in orther to make it a viable alternative of renewable energy. The present research studied the effect of treated wastewater used as an alternative culture medium for growth and productivity of a Chlamydomonas sp. strain isolated from landfills leachate of a treatment pond located in Southern Brazil. Three culture media were evaluated, the control consisted of synthetic TAP medium, other, consisting of 50% TAP medium and 50% wastewater, and another consisting of 100% wastewater. The growth parameters do not have significant difference among the three culture media. Also, productivity do not have significant difference among the cultures with TAP medium and with 100% wastewater, resulting in dry weight values of 1,4±0,14g/L and 1,3±0,19g/L respectively. The culture with 50% TAP medium and 50% wastewater showed the highest productivity, showing an average dry weight value of 1,7±0,07g/L. The results indicate that treated wastewater can be used as an alternative culture medium for Chlamydomonas sp. strain without negative effects on growth and productivity, and possible leading to a decrease in production costs.

Effect of Temperature and light intensity on growth and Photosynthetic Activity of Chlamydomonas reinhard II

International Nuclear Information System (INIS)

Alfonsel Jaen, M.; Fernandez Gonzalez, J.

1985-01-01

The effect of five temperatures (15,20,25,30 and 35 degree centigree) and two levels of illumination on growth and photosynthetic activity of Chlamydomonas reinhard II has been studied. The growth of the cultures was evaluated by optical density. Photosynthetic activity has been carried out studying either the assimilation rate of C0 2 labelled with C-14 or the oxygen evolution by means of polarographic measurements. The maximum photosynthetic rate has been obtained at 25 degree centigree for the lower level of illumination (2400 lux) and at 35 degree centigree for the higher one (13200 lux) and at 35 degree centigree for the higher ono (13200 lux). These results suggest an interaction of temperature and illumination on photosynthetic activity. (Author) 37 refs

Establishing Chlamydomonas reinhardtii as an industrial biotechnology host.

Science.gov (United States)

Scaife, Mark A; Nguyen, Ginnie T D T; Rico, Juan; Lambert, Devinn; Helliwell, Katherine E; Smith, Alison G

2015-05-01

Microalgae constitute a diverse group of eukaryotic unicellular organisms that are of interest for pure and applied research. Owing to their natural synthesis of value-added natural products microalgae are emerging as a source of sustainable chemical compounds, proteins and metabolites, including but not limited to those that could replace compounds currently made from fossil fuels. For the model microalga, Chlamydomonas reinhardtii, this has prompted a period of rapid development so that this organism is poised for exploitation as an industrial biotechnology platform. The question now is how best to achieve this? Highly advanced industrial biotechnology systems using bacteria and yeasts were established in a classical metabolic engineering manner over several decades. However, the advent of advanced molecular tools and the rise of synthetic biology provide an opportunity to expedite the development of C. reinhardtii as an industrial biotechnology platform, avoiding the process of incremental improvement. In this review we describe the current status of genetic manipulation of C. reinhardtii for metabolic engineering. We then introduce several concepts that underpin synthetic biology, and show how generic parts are identified and used in a standard manner to achieve predictable outputs. Based on this we suggest that the development of C. reinhardtii as an industrial biotechnology platform can be achieved more efficiently through adoption of a synthetic biology approach. © 2015 The Authors The Plant Journal published by Society for Experimental Biology and John Wiley & Sons Ltd.

Optimization of the C11-BODIPY581/591 Dye for the Determination of Lipid Oxidation in Chlamydomonas reinhardtii by Flow Cytometry

OpenAIRE

CHELONI Giulia

2013-01-01

Lipid oxidation is a recognized end point for the study of oxidative stress and is an important parameter to describe the mode of micropollutant action on aquatic microorganisms. Therefore the development of quick and reliable methodologies probing the oxidative stress and damage in living cells is highly sought. In the present proof of concept work we examined the potential of the fluorescent dye C11 BODIPY591/581 to probe lipid oxidation in the green microalga Chlamydomonas reinhardtii. C11...

Copper excess-induced large reversible and small irreversible adaptations in a population of Chlamydomonas reinhardtii CW15 (Chlorophyta

Directory of Open Access Journals (Sweden)

Bartosz Pluciński

2018-03-01

Full Text Available Two Chlamydomonas reinhardtii CW15 populations modified by an excess of copper in growth medium were obtained: a “Cu” population that was continuously grown under the selection pressure of 5 µM Cu2+ (for at least 48 weeks and the “Re” population, where a relatively short (9 week exposure to elevated copper, necessary for acquiring tolerance, was followed by a prolonged period (at least 39 weeks of cultivation at a normal (0.25 µM copper concentration. Cells of the Cu population were able to multiply at a Cu2+ concentration 16 times higher than that of the control population at a normal light intensity and at a Cu2+ concentration 64 times higher when cultivated in dim light. The potential quantum yield of photosystem II (FV/FM ratio under copper stress was also significantly higher for the Cu population than for Re and control populations. The Re population showed only residual tolerance towards the elevated concentration of copper, which is revealed by an FV/FM ratio slightly higher than in the control population under Cu2+ stress in dim light or in darkness. We postulate that in the Chlamydomonas populations studied in this paper, at least two mechanisms of copper tolerance operate. The first mechanism is maintained during cultivation at a standard copper concentration and seems to be connected with photosynthetic apparatus. This mechanism, however, has only low adaptive value under excess of copper. The other mechanism, with a much higher adaptive value, is probably connected with Cu2+ homeostasis at the cellular level, but is lost during cultivation at a normal copper concentration.

A chloroplast pathway for the de novo biosynthesis of triacylglycerol in Chlamydomonas reinhardtii

Energy Technology Data Exchange (ETDEWEB)

Fan, J.; Xu, C.; Andre, C.

2011-06-23

Neutral lipid metabolism has been extensively studied in yeast, plants and mammals. In contrast, little information is available regarding the biochemical pathway, enzymes and regulatory factors involved in the biosynthesis of triacylglycerol (TAG) in microalgae. In the conventional TAG biosynthetic pathway widely accepted for yeast, plants and mammals, TAG is assembled in the endoplasmic reticulum (ER) from its immediate precursor diacylglycerol (DAG) made by ER-specific acyltransferases, and is deposited exclusively in lipid droplets in the cytosol. Here, we demonstrated that the unicellular microalga Chlamydomonas reinhardtii employs a distinct pathway that uses DAG derived almost exclusively from the chloroplast to produce TAG. This unique TAG biosynthesis pathway is largely dependent on de novo fatty acid synthesis, and the TAG formed in this pathway is stored in lipid droplets in both the chloroplast and the cytosol. These findings have wide implications for understanding TAG biosynthesis and storage and other areas of lipid metabolism in microalgae and other organisms.

High-level 13C-enrichment of random and synchronous populations of Chlamydomonas reinhardii

International Nuclear Information System (INIS)

Price, R.L.; Crissman, H.A.; Martin, J.C.; Kollman, V.H.

1975-01-01

The alga Chlamydomonas reinhardii was grown in suspension culture at high levels of 13 C-enrichment (98 mol percent) both in synchronous and random populations for the purpose of investigating possible macro- and ultrastructural changes in the cell as induced by essentially total carbon replacement. The algae, grown in spinner flasks, were analyzed using a newly developed multiparameter flow-system technique applied to characterizing various algal genera. The versatility of this technique provides for measuring and processing several cell characteristics simultaneously and separating cells according to selected combinations of parameters. In these studies, cell volume (by Coulter aperture) and DNA and chlorophyll content were determined simultaneously. Cell ultrastructure was examined at various levels of isotope enrichment and time periods by electron microscopy. The data presented for synchronous growth of this organism demonstrate the absence of biological effects (considering the parameters measured) due to the almost total replacement of cellular 12 C with 13 C. Interpretational problems encountered when looking for biological effects on random populations are discussed

Experimental evolution of an alternating uni- and multicellular life cycle in Chlamydomonas reinhardtii

Science.gov (United States)

Ratcliff, William C.; Herron, Matthew D.; Howell, Kathryn; Pentz, Jennifer T.; Rosenzweig, Frank; Travisano, Michael

2013-01-01

The transition to multicellularity enabled the evolution of large, complex organisms, but early steps in this transition remain poorly understood. Here we show that multicellular complexity, including development from a single cell, can evolve rapidly in a unicellular organism that has never had a multicellular ancestor. We subject the alga Chlamydomonas reinhardtii to conditions that favour multicellularity, resulting in the evolution of a multicellular life cycle in which clusters reproduce via motile unicellular propagules. While a single-cell genetic bottleneck during ontogeny is widely regarded as an adaptation to limit among-cell conflict, its appearance very early in this transition suggests that it did not evolve for this purpose. Instead, we find that unicellular propagules are adaptive even in the absence of intercellular conflict, maximizing cluster-level fecundity. These results demonstrate that the unicellular bottleneck, a trait essential for evolving multicellular complexity, can arise rapidly via co-option of the ancestral unicellular form. PMID:24193369

Ultraviolet radiation and the snow alga Chlamydomonas nivalis (Bauer) Wille.

Science.gov (United States)

Gorton, Holly L; Vogelmann, Thomas C

2003-06-01

Aplanospores of Chlamydomonas nivalis are frequently found in high-altitude, persistent snowfields where they are photosynthetically active despite cold temperatures and high levels of visible and ultraviolet (UV) radiation. The goals of this work were to characterize the UV environment of the cells in the snow and to investigate the existence and localization of screening compounds that might prevent UV damage. UV irradiance decreased precipitously in snow, with UV radiation of wavelengths 280-315 nm and UV radiation of wavelengths 315-400 nm dropping to 50% of incident levels in the top 1 and 2 cm, respectively. Isolated cell walls exhibited UV absorbance, possibly by sporopollenin, but this absorbance was weak in images of broken or plasmolyzed cells observed through a UV microscope. The cells also contained UV-absorbing cytoplasmic compounds, with the extrachloroplastic carotenoid astaxanthin providing most of the screening. Additional screening compound(s) soluble in aqueous methanol with an absorption maximum at 335 nm played a minor role. Thus, cells are protected against potentially high levels of UV radiation by the snow itself when they live several centimeters beneath the surface, and they rely on cellular screening compounds, chiefly astaxanthin, when located near the surface where UV fluxes are high.

Systems-Wide Analysis of Acclimation Responses to Long-Term Heat Stress and Recovery in the Photosynthetic Model Organism Chlamydomonas reinhardtii[W][OPEN

Science.gov (United States)

Hemme, Dorothea; Veyel, Daniel; Mühlhaus, Timo; Sommer, Frederik; Jüppner, Jessica; Unger, Ann-Katrin; Sandmann, Michael; Fehrle, Ines; Schönfelder, Stephanie; Steup, Martin; Geimer, Stefan; Kopka, Joachim; Giavalisco, Patrick; Schroda, Michael

2014-01-01

We applied a top-down systems biology approach to understand how Chlamydomonas reinhardtii acclimates to long-term heat stress (HS) and recovers from it. For this, we shifted cells from 25 to 42°C for 24 h and back to 25°C for ≥8 h and monitored abundances of 1856 proteins/protein groups, 99 polar and 185 lipophilic metabolites, and cytological and photosynthesis parameters. Our data indicate that acclimation of Chlamydomonas to long-term HS consists of a temporally ordered, orchestrated implementation of response elements at various system levels. These comprise (1) cell cycle arrest; (2) catabolism of larger molecules to generate compounds with roles in stress protection; (3) accumulation of molecular chaperones to restore protein homeostasis together with compatible solutes; (4) redirection of photosynthetic energy and reducing power from the Calvin cycle to the de novo synthesis of saturated fatty acids to replace polyunsaturated ones in membrane lipids, which are deposited in lipid bodies; and (5) when sinks for photosynthetic energy and reducing power are depleted, resumption of Calvin cycle activity associated with increased photorespiration, accumulation of reactive oxygen species scavengers, and throttling of linear electron flow by antenna uncoupling. During recovery from HS, cells appear to focus on processes allowing rapid resumption of growth rather than restoring pre-HS conditions. PMID:25415976

The TOR Signaling Network in the Model Unicellular Green Alga Chlamydomonas reinhardtii

Directory of Open Access Journals (Sweden)

María Esther Pérez-Pérez

2017-07-01

Full Text Available Cell growth is tightly coupled to nutrient availability. The target of rapamycin (TOR kinase transmits nutritional and environmental cues to the cellular growth machinery. TOR functions in two distinct multiprotein complexes, termed TOR complex 1 (TORC1 and TOR complex 2 (TORC2. While the structure and functions of TORC1 are highly conserved in all eukaryotes, including algae and plants, TORC2 core proteins seem to be missing in photosynthetic organisms. TORC1 controls cell growth by promoting anabolic processes, including protein synthesis and ribosome biogenesis, and inhibiting catabolic processes such as autophagy. Recent studies identified rapamycin-sensitive TORC1 signaling regulating cell growth, autophagy, lipid metabolism, and central metabolic pathways in the model unicellular green alga Chlamydomonas reinhardtii. The central role that microalgae play in global biomass production, together with the high biotechnological potential of these organisms in biofuel production, has drawn attention to the study of proteins that regulate cell growth such as the TOR kinase. In this review we discuss the recent progress on TOR signaling in algae.

The TOR Signaling Network in the Model Unicellular Green Alga Chlamydomonas reinhardtii.

Science.gov (United States)

Pérez-Pérez, María Esther; Couso, Inmaculada; Crespo, José L

2017-07-12

Cell growth is tightly coupled to nutrient availability. The target of rapamycin (TOR) kinase transmits nutritional and environmental cues to the cellular growth machinery. TOR functions in two distinct multiprotein complexes, termed TOR complex 1 (TORC1) and TOR complex 2 (TORC2). While the structure and functions of TORC1 are highly conserved in all eukaryotes, including algae and plants, TORC2 core proteins seem to be missing in photosynthetic organisms. TORC1 controls cell growth by promoting anabolic processes, including protein synthesis and ribosome biogenesis, and inhibiting catabolic processes such as autophagy. Recent studies identified rapamycin-sensitive TORC1 signaling regulating cell growth, autophagy, lipid metabolism, and central metabolic pathways in the model unicellular green alga Chlamydomonas reinhardtii . The central role that microalgae play in global biomass production, together with the high biotechnological potential of these organisms in biofuel production, has drawn attention to the study of proteins that regulate cell growth such as the TOR kinase. In this review we discuss the recent progress on TOR signaling in algae.

Hydrogen production by Chlamydomonas reinhardtii under light driven sulfur deprived condition

Energy Technology Data Exchange (ETDEWEB)

Vijayaraghavan, Krishnan; Karthik, Rajendran [Biotechnology Research Division, Department of Biotechnology, Prathyusha Institute of Technology and Management, Aranvoyalkuppam, Thiruvallur District 602025, Tamil Nadu (India); Kamala Nalini, S.P. [Department of Biotechnology, Vel Group of Educational Institutions, Avadi, Alamadhi Road, Chennai 600062, Tamil Nadu (India)

2009-10-15

This article explores the possibility of demonstrating sustainable photohydrogen production using Chlamydomonas reinhardtii when grown in sulfur deprived photoautotrophic condition. The hydrogen evolving capability of the algal species was monitored based on alternating light and dark period. Investigation was carried out during the day time in order to exploit the solar energy for meeting the demand of the light period. The results showed that when the reactor was operated at varying photoperiod namely 2, 3 and 4 h of alternating light and dark period, the gas generation was found to be 32 {+-} 4, 63 {+-} 7 and 52 {+-} 5 mL/h, while the corresponding hydrogen content was 47, 86 and 87% respectively. Functional components of hydrogen generation reaction centers were also analyzed, which showed that the PS(I) reaction centers were involved in hydrogen production pathway, as the light absorption by PS(I) was prerequisite for hydrogen generation under sulfur deprived photoautotrophic condition. The findings showed a higher gas yield and hydrogen content under dark period, whereas under light period the gas content was below detectable level for hydrogen due to the reversible hydrogenase reaction. (author)

Nickel and low CO2-controlled motility in Chlamydomonas through complementation of a paralyzed flagella mutant with chemically regulated promoters

Directory of Open Access Journals (Sweden)

Rosenbaum Joel L

2011-01-01

Full Text Available Abstract Background Chlamydomonas reinhardtii is a model system for the biology of unicellular green algae. Chemically regulated promoters, such as the nickel-inducible CYC6 or the low CO2-inducible CAH1 promoter, may prove useful for expressing, at precise times during its cell cycle, proteins with relevant biological functions, or complementing mutants in genes encoding such proteins. To this date, this has not been reported for the above promoters. Results We fused the CYC6 and CAH1 promoters to an HA-tagged RSP3 gene, encoding a protein of the flagellar radial spoke complex. The constructs were used for chemically regulated complementation of the pf14 mutant, carrying an ochre mutation in the RSP3 gene. 7 to 8% of the transformants showed cells with restored motility after induction with nickel or transfer to low CO2 conditions, but not in non-inducing conditions. Maximum complementation (5% motile cells was reached with very different kinetics (5-6 hours for CAH1, 48 hours for CYC6. The two inducible promoters drive much lower levels of RSP3 protein expression than the constitutive PSAD promoter, which shows almost complete rescue of motility. Conclusions To our knowledge, this is the first example of the use of the CYC6 or CAH1 promoters to perform a chemically regulated complementation of a Chlamydomonas mutant. Based on our data, the CYC6 and CAH1 promoters should be capable of fully complementing mutants in genes whose products exert their biological activity at low concentrations.

Screening of Chlamydomonas reinhardtii Populations with Single-Cell Resolution by Using a High-Throughput Microscale Sample Preparation for Matrix-Assisted Laser Desorption Ionization Mass Spectrometry.

Science.gov (United States)

Krismer, Jasmin; Sobek, Jens; Steinhoff, Robert F; Fagerer, Stephan R; Pabst, Martin; Zenobi, Renato

2015-08-15

The consequences of cellular heterogeneity, such as biocide persistence, can only be tackled by studying each individual in a cell population. Fluorescent tags provide tools for the high-throughput analysis of genomes, RNA transcripts, or proteins on the single-cell level. However, the analysis of lower-molecular-weight compounds that elude tagging is still a great challenge. Here, we describe a novel high-throughput microscale sample preparation technique for single cells that allows a mass spectrum to be obtained for each individual cell within a microbial population. The approach presented includes spotting Chlamydomonas reinhardtii cells, using a noncontact microarrayer, onto a specialized slide and controlled lysis of cells separated on the slide. Throughout the sample preparation, analytes were traced and individual steps optimized using autofluorescence detection of chlorophyll. The lysates of isolated cells are subjected to a direct, label-free analysis using matrix-assisted laser desorption ionization mass spectrometry. Thus, we were able to differentiate individual cells of two Chlamydomonas reinhardtii strains based on single-cell mass spectra. Furthermore, we showed that only population profiles with real single-cell resolution render a nondistorted picture of the phenotypes contained in a population. Copyright © 2015, American Society for Microbiology. All Rights Reserved.

Kinetic modeling of light limitation and sulfur deprivation effects in the induction of hydrogen production with Chlamydomonas reinhardtii: Part I. Model development and parameter identification.

Science.gov (United States)

Fouchard, Swanny; Pruvost, Jérémy; Degrenne, Benoit; Titica, Mariana; Legrand, Jack

2009-01-01

Chlamydomonas reinhardtii is a green microalga capable of turning its metabolism towards H2 production under specific conditions. However this H2 production, narrowly linked to the photosynthetic process, results from complex metabolic reactions highly dependent on the environmental conditions of the cells. A kinetic model has been developed to relate culture evolution from standard photosynthetic growth to H2 producing cells. It represents transition in sulfur-deprived conditions, known to lead to H2 production in Chlamydomonas reinhardtii, and the two main processes then induced which are an over-accumulation of intracellular starch and a progressive reduction of PSII activity for anoxia achievement. Because these phenomena are directly linked to the photosynthetic growth, two kinetic models were associated, the first (one) introducing light dependency (Haldane type model associated to a radiative light transfer model), the second (one) making growth a function of available sulfur amount under extracellular and intracellular forms (Droop formulation). The model parameters identification was realized from experimental data obtained with especially designed experiments and a sensitivity analysis of the model to its parameters was also conducted. Model behavior was finally studied showing interdependency between light transfer conditions, photosynthetic growth, sulfate uptake, photosynthetic activity and O2 release, during transition from oxygenic growth to anoxic H2 production conditions.

Ultraviolet modification of Chlamydomonas reinhardtii for carbon capture

Directory of Open Access Journals (Sweden)

Gopal NS

2016-04-01

Full Text Available Nikhil S Gopal,1 K Sudhakar2 1The Lawrenceville School, Lawrenceville, NJ, USA; 2Bioenergy Laboratory, Malauna Azad National Institute of Technology, Bhopal, India Purpose: Carbon dioxide (CO2 levels have been rising rapidly. Algae are single-cell organisms with highly efficient CO2 uptake mechanisms. Algae yield two to ten times more biomass versus terrestrial plants and can grow nearly anywhere. Large scale CO2 sequestration is not yet sustainable due to high amounts of nitrogen (N and phosphate (P needed to grow algae in media. Methods: Mutant strains of Chlamydomonas reinhardtii were created using ultraviolet light (2.2–3 K J/m2 and natural selection using media with 20%–80% lower N and P compared to standard Sueoka's high salt medium. Strains were selected based upon growth in media concentrations varying from 20% to 80% less N/P compared to control. Biomass was compared to wild-type control (CC-125 using direct counts, optical density dry weight, and mean doubling time. Results: Mean doubling time was 20 and 25 hours in the low N and N/P strains, respectively (vs 66 hours in control. Using direct counts, growth rates of mutant strains of low N and N/P cultures were not statistically different from control (P=0.37 and 0.70, respectively. Conclusion: Two new strains of algae, as well as wild-type control, were able to grow while using 20%–40% less N and P. Ultraviolet light-based modification of algae is an inexpensive and alternative option to genetic engineering techniques. This technique might make larger scale biosequestration possible. Keywords: biosequestration, ultraviolet, carbon sequestration, carbon capture, algae

An omics based assessment of cadmium toxicity in the green alga Chlamydomonas reinhardtii

Energy Technology Data Exchange (ETDEWEB)

Jamers, An; Blust, Ronny; De Coen, Wim [Laboratory for Ecophysiology, Biochemistry and Toxicology, Department of Biology, University of Antwerp, Groenenborgerlaan 171, 2020 Antwerp (Belgium); Griffin, Julian L. [Department of Biochemistry, University of Cambridge, 80 Tennis Court Road, Cambridge CB2 2QA (United Kingdom); Jones, Oliver A.H., E-mail: oliver.jones@rmit.edu.au [School of Applied Sciences, RMIT University, GPO Box 2476, Melbourne, VIC 3001 (Australia)

2013-01-15

The effects of cadmium were assessed in the freshwater alga Chlamydomonas reinhardtii. Algae were exposed to concentrations of 0, 8.1 or 114.8 {mu}M of cadmium and growth rates, gene transcription and metabolite profiles were examined after 48 and 72 h of exposure. In algae exposed to 8.1 {mu}M Cd, several genes were differentially transcribed after 48 h but no adverse growth related effects were detected. A transient effect on both gene transcription patterns and metabolite profiles could be discerned after 48 h of exposure but the majority of these changes disappeared after 72 h. In contrast, all effects were more pronounced at the 114.8 {mu}M cadmium exposure. Here growth was clearly reduced and transcription of a large number of genes involved in oxidative stress defense mechanisms was differentially increased. Metabolites involved in the glutathione synthesis pathway (an important antioxidant defense) were also affected but the effects of cadmium were found to be more pronounced at the transcript level than in the metabolome, suggesting that the former exhibits greater sensitivity toward cadmium exposure.

The two parallel photocycles of the Chlamydomonas sensory photoreceptor histidine kinase rhodopsin 1.

Science.gov (United States)

Luck, Meike; Hegemann, Peter

2017-10-01

Histidine kinase rhodopsins (HKRs) belong to a class of unexplored sensory photoreceptors that share a similar modular architecture. The light sensing rhodopsin domain is covalently linked to signal-transducing modules and in some cases to a C-terminal guanylyl-cyclase effector. In spite of their wide distribution in unicellular organisms, very little is known about their physiological role and mechanistic functioning. We investigated the photochemical properties of the recombinant rhodopsin-fragment of Cr-HKR1 originating from Chlamydomonas reinhardtii. Our spectroscopic studies revealed an unusual thermal stability of the photoproducts with the deprotonated retinal Schiff base (RSB). Upon UV-irradiation these Rh-UV states with maximal absorbance in the UVA-region (Rh-UV) photochemically convert to stable blue light absorbing rhodopsin (Rh-Bl) with protonated chromophore. The heterogeneity of the sample is based on two parallel photocycles with the chromophore in C 15 =N-syn- or -anti-configuration. This report represents an attempt to decipher the underlying reaction schemes and interconversions of the two coexisting photocycles. Copyright © 2017 Elsevier GmbH. All rights reserved.

The effect of caffeine on repair in chlamydomonas reinhardtii. Pt. 1

International Nuclear Information System (INIS)

Rosen, H.; Rehn, M.M.; Johnson, B.A.

1980-01-01

The effect of caffeine on repair was studied in the green alga Chlamydomonas reinhardtii. Treatment of UV-irradiated wild-type (UVS + ) cells with a sublethal level of caffeine caused a significant increase in survival compared to untreated UV-irradiated cells. Caffeine did not affect survival in the repair-deficient strain UVSE1, which is deficient in repair of UV-induced damage carried out by enzymes associated with recombination during meiosis. A significant increase in survival in the presence of caffeine was observed in the repair-deficient strain UVSE4 in which recombination during meiosis is not affected. Treatment of zygotes homozygous for UVS + , UVSE1, or UVSE4 with sublethal levels of caffeine caused marked increases in recombination frequency in UVS + and UVSE4 zygotes and no increase in recombination in UVSE1 zygotes. These results indicate that caffeine increases recombination in normal strains. Increased opportunity for recombination caused by caffeine would not result in increased recombination frequency in the UVSE1 strain, assuming limited-recombination enzyme activity in this strain. The observed increase in survival following UV-irradiation in the presence of caffeine in strains having normal recombination would therefore be associated with a caffeine-induced increase in opportunities for recombination repair. (orig.)

Efficient expression of green fluorescent protein (GFP) mediated by a chimeric promoter in Chlamydomonas reinhardtii

Science.gov (United States)

Wu, Jinxia; Hu, Zhangli; Wang, Chaogang; Li, Shuangfei; Lei, Anping

2008-08-01

To improve the expression efficiency of exogenous genes in Chlamydomonas reinhardtii, a high efficient expression vector was constructed. Green fluorescent protein (GFP) was expressed in C. reinhardtii under the control of promoters: RBCS2 and HSP70A-RBCS2. Efficiency of transformation and expression were compared between two transgenic algae: RBCS2 mediated strain Tran-I and HSP70A-RBCS2 mediated strain Tran-II. Results show that HSP70A-RBCS2 could improve greatly the transformation efficiency by approximately eightfold of RBCS2, and the expression efficiency of GFP in Tran-II was at least double of that in Tran-I. In addition, a threefold increase of GFP in Tran-II was induced by heat shock at 40°C. All of the results demonstrated that HSP70A-RBCS2 was more efficient than RBCS2 in expressing exogenous gene in C. reinhardtii.

Shewanella oneidensis: a new and efficient System for Expression and Maturation of heterologous [Fe-Fe] Hydrogenase from Chlamydomonas reinhardtii

Directory of Open Access Journals (Sweden)

Sybirna Kateryna

2008-09-01

Full Text Available Abstract Background The eukaryotic green alga, Chlamydomonas reinhardtii, produces H2 under anaerobic conditions, in a reaction catalysed by a [Fe-Fe] hydrogenase HydA1. For further biochemical and biophysical studies a suitable expression system of this enzyme should be found to overcome its weak expression in the host organism. Two heterologous expression systems used up to now have several advantages. However they are not free from some drawbacks. In this work we use bacterium Shewanella oneidensis as a new and efficient system for expression and maturation of HydA1 from Chlamydomonas reinhardtii. Results Based on codon usage bias and hydrogenase maturation ability, the bacterium S. oneidensis, which possesses putative [Fe-Fe] and [Ni-Fe] hydrogenase operons, was selected as the best potential host for C. reinhardtii [Fe-Fe] hydrogenase expression. Hydrogen formation by S. oneidensis strain AS52 (ΔhydAΔhyaB transformed with a plasmid bearing CrHydA1 and grown in the presence of six different substrates for anaerobic respiration was determined. A significant increase in hydrogen evolution was observed for cells grown in the presence of trimethylamine oxide, dimethylsulfoxide and disodium thiosulfate, showing that the system of S. oneidensis is efficient for heterologous expression of algal [Fe-Fe] hydrogenase. Conclusion In the present work a new efficient system for heterologous expression and maturation of C. reinhardtii hydrogenase has been developed. HydA1 of C. reinhardtii was purified and shown to contain 6 Fe atoms/molecule of protein, as expected. Using DMSO, TMAO or thiosulfate as substrates for anaerobic respiration during the cell growth, 0.4 – 0.5 mg l-1(OD600 = 1 of catalytically active HydA1 was obtained with hydrogen evolution rate of ~700 μmol H2 mg-1 min-1.

Rubisco mutants of Chlamydomonas reinhardtii enhance photosynthetic hydrogen production.

Science.gov (United States)

Pinto, T S; Malcata, F X; Arrabaça, J D; Silva, J M; Spreitzer, R J; Esquível, M G

2013-06-01

Molecular hydrogen (H2) is an ideal fuel characterized by high enthalpy change and lack of greenhouse effects. This biofuel can be released by microalgae via reduction of protons to molecular hydrogen catalyzed by hydrogenases. The main competitor for the reducing power required by the hydrogenases is the Calvin cycle, and rubisco plays a key role therein. Engineered Chlamydomonas with reduced rubisco levels, activity and stability was used as the basis of this research effort aimed at increasing hydrogen production. Biochemical monitoring in such metabolically engineered mutant cells proceeded in Tris/acetate/phosphate culture medium with S-depletion or repletion, both under hypoxia. Photosynthetic activity, maximum photochemical efficiency, chlorophyll and protein levels were all measured. In addition, expression of rubisco, hydrogenase, D1 and Lhcb were investigated, and H2 was quantified. At the beginning of the experiments, rubisco increased followed by intense degradation. Lhcb proteins exhibited monomeric isoforms during the first 24 to 48 h, and D1 displayed sensitivity under S-depletion. Rubisco mutants exhibited a significant decrease in O2 evolution compared with the control. Although the S-depleted medium was much more suitable than its complete counterpart for H2 production, hydrogen release was observed also in sealed S-repleted cultures of rubisco mutated cells under low-moderate light conditions. In particular, the rubisco mutant Y67A accounted for 10-15-fold higher hydrogen production than the wild type under the same conditions and also displayed divergent metabolic parameters. These results indicate that rubisco is a promising target for improving hydrogen production rates in engineered microalgae.

Radiocesium bioaccumulation in freshwater plankton: Influences of cation concentrations (K{sup +} and Na{sup +}) on direct uptake of {sup 137}Cs in Chlamydomonas, Scenedesmus and Daphnia. Food-chain transfer of {sup 137}Cs from Chlamydomonas to Daphnia at different K{sup +} concentrations

Energy Technology Data Exchange (ETDEWEB)

Hagstroem, J. [Uppsala Univ., Dept. of Limnology, Uppsala (Sweden)

2002-04-01

The influences of cation concentrations (K{sup +} and Na{sup +}) on radiocesium ({sup 137}Cs) bioaccumulation in two freshwater phytoplankton species (Scenedesmus quadricauda and Chlamydomonas noctigama) were systematically investigated in batch-cultures monitored during two weeks. Both species were cultured at 9 {mu}E M{sup -2} s{sup -1} constant illumination at 20 deg. C. The exponential growth phase lasted for more than 100 hours ({mu} {approx_equal} 0.02 h{sup -1} for C. noctigama and 0.03 h{sup -1} for S, quadricauda). Over cation concentration ranges encountered in natural fresh waters ([K{sup +}] from 0.1 {mu}M to 3 mM, [Na{sup +}] from 20 {mu}M to 3 mM), a more than three order of magnitude variation was found for both intake rate and observed bioconcentration factors (BCF) at apparent steady-state (from less than 10{sup 3} to 10{sup 6} L (kg C){sup -1}). For both species, the major effector on BCF and uptake rate was external [K{sup +}], which was inversely proportional to these parameters over wide ranges (1-1000 {mu}M for S. quadricauda and 0.1 to 300 {mu}M for C. noctigama). At concentrations above these ranges K{sup +} still reduced {sup 137} Cs bio-uptake, but less effectively. A minor influence of external [Na{sup +}] on {sup 137}Cs bioaccumulation was indicated for S. quadricauda, whereas no such influence was significant for C. noctigama. A biphasic pattern for {sup 137}Cs bioaccumulation was discovered in C. noctigama. A rapid 'quasi-steady state' with an effective equilibration time of less than 100 hours was approached during the exponential growth phase. A surge in the uptake occurred when exponential growth ceased, and this pattern was consistent over the range 30 {mu}M to 1.4 mM external [K{sup +}]. Since depletion of external [K{sup +}] was not detected for these treatments, this pattern can only be explained if there are at least two different cellular compartments involved. Although less certain, a second steady-state BCF

Águas com predominância de Eutreptia lanowi steuer e Chlamydomonas reinhardi dangeard no plancton, na enseada de Inhauma, Baía de Guanabara

Directory of Open Access Journals (Sweden)

Lejeune P. H. de Oliveira

1962-03-01

Full Text Available In brackish waters of a creek of Guanabara Bay, the author points by the first time the presence of Chlamydomonas reinhardi, Eutreptia lanowi, Oscillatoria putrida, O. limosa, O. chlorina that were unknown in our waters; such biologic indicators proved themselves pollutional conditions, so bad a stark-mesosaprobic regime. Other news are plankton analysis by the Standar methods, of two most expressive samples of water masses;also the mobility of the plankters are measured in micra by second.

The energy balance of the biomass generation of Chlamydomonas acidophila under acidic and neutral conditions and Chlamydomonas reinhardtii; Die Energiebilanz der Biomasseneubildung von Chlamydomonas acidophila unter sauren und neutralen Bedingungen und von Chlamydomonas reinhardtii

Energy Technology Data Exchange (ETDEWEB)

Langner, Uwe

2009-01-16

In this study the influence of pH < 3 as an extreme environment has been investigated for the eukaryotic green alga Chlamydomonas (C.) acidophila. The limited number of trophic levels, consisting of bacteria, phytoplankton, zooplankton and macrophytes, is a special characteristic of extreme acidic water bodies. C. acidophila was isolated from an extreme acidic mining lake (RL 111) (Bissinger et al. 2000). A special feature of the examined algal species is its wide tolerance range of external pH values from 2 to 7 (Cassin 1874, Gerloff-Elias et al. 2005a). C. acidophila is a dominant species in the acidic mining lakes, it can grow up to chlorophyll maxima of 500 {mu}g L{sup -1} during the summer time (Nixdorf et al. 1998, 2003). The alga can be found elsewhere in extreme acidic water bodies around the world. The hydrochemistry of the acidic mining lakes in the central regions of Germany and Lusatia show clear differences compared to neutral water bodies. Some of the characteristics of acidic mining lakes are high metal and heavy metal (aluminum) concentrations, high ion concentrations, which lead to high conductivity, as well as low phosphate concentrations, ammonium as the only nitrogen source and CO{sub 2} as the only inorganic carbon source. Many eukaryotic microalgae in acidic water bodies including C. acidophila show a neutral cytosolic pH. This is provided by special adaptations of the organisms. Thus, organisms in extreme acidic environments have a positive cell surface charge, a very efficient H{sup +}-ATPase and high internal buffer capacities. These mechanisms work contrary to the proton influx and the acidification of the cytosol and are therefore proof for the physiological impact of osmoregulation by microalgae in extreme acidic environments (Sekler et al.1991, Weiss et al. 1999). Hence, these mechanisms also complicate the access to nutrients for the algal cell. The passive influx of positively charged ions such as potassium or ammonium is reduced by

Role of metal mixtures (Ca, Cu and Pb) on Cd bioaccumulation and phytochelatin production by Chlamydomonas reinhardtii.

Science.gov (United States)

Abboud, Pauline; Wilkinson, Kevin J

2013-08-01

The goal of the study was to determine whether metal uptake and biological effects could be predicted by free ion concentrations when organisms were exposed to Cd and a second metal. Bioaccumulation and algal phytochelatin (PC) concentrations were determined for Chlamydomonas reinhardtii following a 6-h exposure. Bioaccumulation results, after six hours of exposure, showed that Cd uptake decreased in the presence of relatively high concentrations of Ca, Cu and Pb, however, Cd bioaccumulation increased in the presence of ca. equimolar concentrations of Cu. A good correlation was observed between the production of PCs and the amount of metals bioaccumulated for the binary mixtures of Cd-Pb and Cd-Cu, but not the Cd-Ca mixture. Overall, the results suggested that, in the case of mixtures, bioaccumulated metal rather than free ion concentrations would be a better predictor of biological effect. Copyright © 2013 Elsevier Ltd. All rights reserved.

Flagellar coordination in Chlamydomonas cells held on micropipettes.

Science.gov (United States)

Rüffer, U; Nultsch, W

1998-01-01

The two flagella of Chlamydomonas are known to beat synchronously: During breaststroke beating they are generally coordinated in a bilateral way while in shock responses during undulatory beating coordination is mostly parallel [Rüffer and Nultsch, 1995: Botanica Acta 108:169-276]. Analysis of a great number of shock responses revealed that in undulatory beats also periods of bilateral coordination are found and that the coordination type may change several times during a shock response, without concomitant changes of the beat envelope and the beat period. In normal wt cells no coordination changes are found during breaststroke beating, but only short temporary asynchronies: During 2 or 3 normal beats of the cis flagellum, the trans flagellum performs 3 or 4 flat beats with a reduced beat envelope and a smaller beat period, resulting in one additional trans beat. Long periods with flat beats of the same shape and beat period are found in both flagella of the non-phototactic mutant ptx1 and in defective wt 622E cells. During these periods, the coordination is parallel, the two flagella beat alternately. A correlation between normal asynchronous trans beats and the parallel-coordinated beats in the presumably cis defective cells and also the undulatory beats is discussed. In the cis defective cells, a perpetual spontaneous change between parallel beats with small beat periods (higher beat frequency) and bilateral beats with greater beat periods (lower beat frequency) are observed and render questionable the existence of two different intrinsic beat frequencies of the two flagella cis and trans. Asynchronies occur spontaneously but may also be induced by light changes, either step-up or step-down, but not by both stimuli in turn as breaststroke flagellar photoresponses (BFPRs). Asynchronies are not involved in phototaxis. They are independent of the BFPRs, which are supposed to be the basis of phototaxis. Both types of coordination must be assumed to be regulated

Chlamydomonas outer arm dynein alters conformation in response to Ca2+.

Science.gov (United States)

Sakato, Miho; Sakakibara, Hitoshi; King, Stephen M

2007-09-01

We have previously shown that Ca(2+) directly activates ATP-sensitive microtubule binding by a Chlamydomonas outer arm dynein subparticle containing the beta and gamma heavy chains (HCs). The gamma HC-associated LC4 light chain is a member of the calmodulin family and binds 1-2 Ca(2+) with K(Ca) = 3 x 10(-5) M in vitro, suggesting it may act as a Ca(2+) sensor for outer arm dynein. Here we investigate interactions between the LC4 light chain and gamma HC. Two IQ consensus motifs for binding calmodulin-like proteins are located within the stem domain of the gamma heavy chain. In vitro experiments indicate that LC4 undergoes a Ca(2+)-dependent interaction with the IQ motif domain while remaining tethered to the HC. LC4 also moves into close proximity of the intermediate chain IC1 in the presence of Ca(2+). The sedimentation profile of the gamma HC subunit changed subtly upon Ca(2+) addition, suggesting that the entire complex had become more compact, and electron microscopy of the isolated gamma subunit revealed a distinct alteration in conformation of the N-terminal stem in response to Ca(2+) addition. We propose that Ca(2+)-dependent conformational change of LC4 has a direct effect on the stem domain of the gamma HC, which eventually leads to alterations in mechanochemical interactions between microtubules and the motor domain(s) of the outer dynein arm.

The Effect of DNA and Sodium Cholate Dispersed Single-Walled Carbon Nano tubes on the Green Algae Chlamydomonas reinhardtii

International Nuclear Information System (INIS)

Williams, R.M.; Cox, Z.; Dolash, B.D.; Sooter, L.J.; Williams, R.M.; Taylor, H.K.; Thomas, J.

2014-01-01

Increasing use of single-walled carbon nano tubes (SWCNTs) will lead to their increased release into the environment. Previous work has shown negative effects of SWCNT on growth and survival of model organisms. The aim of the current study was to determine the effect of SWCNT well-dispersed by either DNA or sodium cholate (SC) on the unicellular green algae Chlamydomonas reinhardtii in stagnant water conditions. Growth measurements were taken up to ten days for algae treated with varied levels of DNA:SWCNT or SC:SWCNT or controls, and chlorophyll content after 10 days was determined. Results show no effect on either growth or chlorophyll content of algae at any concentration or duration. This is in contradiction to prior work showing toxicity of SWCNT to environmental model organisms.

Phytohormone supplementation significantly increases growth of Chlamydomonas reinhardtii cultivated for biodiesel production.

Science.gov (United States)

Park, Won-Kun; Yoo, Gursong; Moon, Myounghoon; Kim, Chul Woong; Choi, Yoon-E; Yang, Ji-Won

2013-11-01

Cultivation is the most expensive step in the production of biodiesel from microalgae, and substantial research has been devoted to developing more cost-effective cultivation methods. Plant hormones (phytohormones) are chemical messengers that regulate various aspects of growth and development and are typically active at very low concentrations. In this study, we investigated the effect of different phytohormones on microalgal growth and biodiesel production in Chlamydomonas reinhardtii and their potential to lower the overall cost of commercial biofuel production. The results indicated that all five of the tested phytohormones (indole-3-acetic acid, gibberellic acid, kinetin, 1-triacontanol, and abscisic acid) promoted microalgal growth. In particular, hormone treatment increased biomass production by 54 to 69 % relative to the control growth medium (Tris-acetate-phosphate, TAP). Phytohormone treatments also affected microalgal cell morphology but had no effect on the yields of fatty acid methyl esters (FAMEs) as a percent of biomass. We also tested the effect of these phytohormones on microalgal growth in nitrogen-limited media by supplementation in the early stationary phase. Maximum cell densities after addition of phytohormones were higher than in TAP medium, even when the nitrogen source was reduced to 40 % of that in TAP medium. Taken together, our results indicate that phytohormones significantly increased microalgal growth, particularly in nitrogen-limited media, and have potential for use in the development of efficient microalgal cultivation for biofuel production.

UV-B photoreceptor-mediated protection of the photosynthetic machinery in Chlamydomonas reinhardtii

Science.gov (United States)

Allorent, Guillaume; Lefebvre-Legendre, Linnka; Chappuis, Richard; Kuntz, Marcel; Truong, Thuy B.; Niyogi, Krishna K.; Goldschmidt-Clermont, Michel

2016-01-01

Life on earth is dependent on the photosynthetic conversion of light energy into chemical energy. However, absorption of excess sunlight can damage the photosynthetic machinery and limit photosynthetic activity, thereby affecting growth and productivity. Photosynthetic light harvesting can be down-regulated by nonphotochemical quenching (NPQ). A major component of NPQ is qE (energy-dependent nonphotochemical quenching), which allows dissipation of light energy as heat. Photodamage peaks in the UV-B part of the spectrum, but whether and how UV-B induces qE are unknown. Plants are responsive to UV-B via the UVR8 photoreceptor. Here, we report in the green alga Chlamydomonas reinhardtii that UVR8 induces accumulation of specific members of the light-harvesting complex (LHC) superfamily that contribute to qE, in particular LHC Stress-Related 1 (LHCSR1) and Photosystem II Subunit S (PSBS). The capacity for qE is strongly induced by UV-B, although the patterns of qE-related proteins accumulating in response to UV-B or to high light are clearly different. The competence for qE induced by acclimation to UV-B markedly contributes to photoprotection upon subsequent exposure to high light. Our study reveals an anterograde link between photoreceptor-mediated signaling in the nucleocytosolic compartment and the photoprotective regulation of photosynthetic activity in the chloroplast. PMID:27930292

UV-B photoreceptor-mediated protection of the photosynthetic machinery in Chlamydomonas reinhardtii.

Science.gov (United States)

Allorent, Guillaume; Lefebvre-Legendre, Linnka; Chappuis, Richard; Kuntz, Marcel; Truong, Thuy B; Niyogi, Krishna K; Ulm, Roman; Goldschmidt-Clermont, Michel

2016-12-20

Life on earth is dependent on the photosynthetic conversion of light energy into chemical energy. However, absorption of excess sunlight can damage the photosynthetic machinery and limit photosynthetic activity, thereby affecting growth and productivity. Photosynthetic light harvesting can be down-regulated by nonphotochemical quenching (NPQ). A major component of NPQ is qE (energy-dependent nonphotochemical quenching), which allows dissipation of light energy as heat. Photodamage peaks in the UV-B part of the spectrum, but whether and how UV-B induces qE are unknown. Plants are responsive to UV-B via the UVR8 photoreceptor. Here, we report in the green alga Chlamydomonas reinhardtii that UVR8 induces accumulation of specific members of the light-harvesting complex (LHC) superfamily that contribute to qE, in particular LHC Stress-Related 1 (LHCSR1) and Photosystem II Subunit S (PSBS). The capacity for qE is strongly induced by UV-B, although the patterns of qE-related proteins accumulating in response to UV-B or to high light are clearly different. The competence for qE induced by acclimation to UV-B markedly contributes to photoprotection upon subsequent exposure to high light. Our study reveals an anterograde link between photoreceptor-mediated signaling in the nucleocytosolic compartment and the photoprotective regulation of photosynthetic activity in the chloroplast.

VU-B radiation inhibits the photosynthetic electron transport chain in chlamydomonas reinhardtii

International Nuclear Information System (INIS)

Cai, W.; Li, X.; Chen, L.

2016-01-01

UV radiation of sunlight is one of harmful factors for earth organisms, especially for photoautotrophs because they require light for energy and biomass production. A number of works have already been done regarding the effects of UV-B radiation at biochemical and molecular level, which showed that UV-B radiation could inhibit photosynthesis activity and reduce photosynthetic electron transport. However quite limited information can accurately make out inhibition site of UV-B radiation on photosynthetic electron transport. In this study, this issue was investigated through measuring oxygen evolution activity, chlorophyll a fluorescence and gene expression in a model unicellular green alga Chlamydomonas reinhardtii. Our results indicated that UV-B radiation could evidently decrease photosynthesis activity and inhibit electron transport by blocking electron transfer process from the first plastoquinone electron acceptors QA to second plastoquinone electron acceptors QB, but not impair electron transfer from the water oxidizing complex to QA. The psbA gene expression was also altered by UV-B radiation, where up-regulation occurred at 2, 4 and 6h after exposure and down-regulation happened at 12 and 24 h after exposure. These results suggested that UV-B could affects D1 protein normal turnover, so there was not enough D1 for binding with QB, which may affect photosynthetic electron transport and photosynthesis activity. (author)

A simple and non-invasive method for nuclear transformation of intact-walled Chlamydomonas reinhardtii.

Directory of Open Access Journals (Sweden)

Sora Kim

Full Text Available Genetic engineering in microalgae is gaining attraction but nuclear transformation methods available so far are either inefficient or require special equipment. In this study, we employ positively charged nanoparticles, 3-aminopropyl-functionalized magnesium phyllosilicate (aminoclay, approximate unit cell composition of [H2N(CH23]8Si8Mg6O12(OH4, for nuclear transformation into eukaryotic microalgae. TEM and EDX analysis of the process of transformation reveals that aminoclay coats negatively-charged DNA biomolecules and forms a self-assembled hybrid nanostructure. Subsequently, when this nanostructure is mixed with microalgal cells and plated onto selective agar plates with high friction force, cell wall is disrupted facilitating delivery of plasmid DNA into the cell and ultimately to the nucleus. This method is not only simple, inexpensive, and non-toxic to cells but also provides efficient transformation (5.03×10(2 transformants/µg DNA, second only to electroporation which needs advanced instrumentation. We present optimized parameters for efficient transformation including pre-treatment, friction force, concentration of foreign DNA/aminoclay, and plasticity of agar plates. It is also confirmed the successful integration and stable expression of foreign gene in Chlamydomonas reinhardtii through molecular methods.

Transcriptome Analysis of Manganese-deficient Chlamydomonas reinhardtii Provides Insight on the Chlorophyll Biosynthesis Pathway

Energy Technology Data Exchange (ETDEWEB)

Lockhart, Ainsley; Zvenigorodsky, Natasha; Pedraza, Mary Ann; Lindquist, Erika

2011-08-11

The biosynthesis of chlorophyll and other tetrapyrroles is a vital but poorly understood process. Recent genomic advances with the unicellular green algae Chlamydomonas reinhardtii have created opportunity to more closely examine the mechanisms of the chlorophyll biosynthesis pathway via transcriptome analysis. Manganese is a nutrient of interest for complex reactions because of its multiple stable oxidation states and role in molecular oxygen coordination. C. reinhardtii was cultured in Manganese-deplete Tris-acetate-phosphate (TAP) media for 24 hours and used to create cDNA libraries for sequencing using Illumina TruSeq technology. Transcriptome analysis provided intriguing insight on possible regulatory mechanisms in the pathway. Evidence supports similarities of GTR (Glutamyl-tRNA synthase) to its Chlorella vulgaris homolog in terms of Mn requirements. Data was also suggestive of Mn-related compensatory up-regulation for pathway proteins CHLH1 (Manganese Chelatase), GUN4 (Magnesium chelatase activating protein), and POR1 (Light-dependent protochlorophyllide reductase). Intriguingly, data suggests possible reciprocal expression of oxygen dependent CPX1 (coproporphyrinogen III oxidase) and oxygen independent CPX2. Further analysis using RT-PCR could provide compelling evidence for several novel regulatory mechanisms in the chlorophyll biosynthesis pathway.

Robust Microplate-Based Methods for Culturing and in Vivo Phenotypic Screening of Chlamydomonas reinhardtii

Directory of Open Access Journals (Sweden)

Timothy C. Haire

2018-03-01

Full Text Available Chlamydomonas reinhardtii (Cr, a unicellular alga, is routinely utilized to study photosynthetic biochemistry, ciliary motility, and cellular reproduction. Its minimal culture requirements, unicellular morphology, and ease of transformation have made it a popular model system. Despite its relatively slow doubling time, compared with many bacteria, it is an ideal eukaryotic system for microplate-based studies utilizing either, or both, absorbance as well as fluorescence assays. Such microplate assays are powerful tools for researchers in the areas of toxicology, pharmacology, chemical genetics, biotechnology, and more. However, while microplate-based assays are valuable tools for screening biological systems, these methodologies can significantly alter the conditions in which the organisms are cultured and their subsequent physiology or morphology. Herein we describe a novel method for the microplate culture and in vivo phenotypic analysis of growth, viability, and photosynthetic pigments of C. reinhardtii. We evaluated the utility of our assay by screening silver nanoparticles for their effects on growth and viability. These methods are amenable to a wide assortment of studies and present a significant advancement in the methodologies available for research involving this model organism.

Alternative photosynthetic electron transport pathways during anaerobiosis in the green alga Chlamydomonas reinhardtii.

Science.gov (United States)

Hemschemeier, Anja; Happe, Thomas

2011-08-01

Oxygenic photosynthesis uses light as energy source to generate an oxidant powerful enough to oxidize water into oxygen, electrons and protons. Upon linear electron transport, electrons extracted from water are used to reduce NADP(+) to NADPH. The oxygen molecule has been integrated into the cellular metabolism, both as the most efficient electron acceptor during respiratory electron transport and as oxidant and/or "substrate" in a number of biosynthetic pathways. Though photosynthesis of higher plants, algae and cyanobacteria produces oxygen, there are conditions under which this type of photosynthesis operates under hypoxic or anaerobic conditions. In the unicellular green alga Chlamydomonas reinhardtii, this condition is induced by sulfur deficiency, and it results in the production of molecular hydrogen. Research on this biotechnologically relevant phenomenon has contributed largely to new insights into additional pathways of photosynthetic electron transport, which extend the former concept of linear electron flow by far. This review summarizes the recent knowledge about various electron sources and sinks of oxygenic photosynthesis besides water and NADP(+) in the context of their contribution to hydrogen photoproduction by C. reinhardtii. This article is part of a Special Issue entitled: Regulation of Electron Transport in Chloroplasts. Copyright © 2011 Elsevier B.V. All rights reserved.

Analysis of motility in multicellular Chlamydomonas reinhardtii evolved under predation.

Directory of Open Access Journals (Sweden)

Margrethe Boyd

Full Text Available The advent of multicellularity was a watershed event in the history of life, yet the transition from unicellularity to multicellularity is not well understood. Multicellularity opens up opportunities for innovations in intercellular communication, cooperation, and specialization, which can provide selective advantages under certain ecological conditions. The unicellular alga Chlamydomonas reinhardtii has never had a multicellular ancestor yet it is closely related to the volvocine algae, a clade containing taxa that range from simple unicells to large, specialized multicellular colonies. Simple multicellular structures have been observed to evolve in C. reinhardtii in response to predation or to settling rate-based selection. Structures formed in response to predation consist of individual cells confined within a shared transparent extracellular matrix. Evolved isolates form such structures obligately under culture conditions in which their wild type ancestors do not, indicating that newly-evolved multicellularity is heritable. C. reinhardtii is capable of photosynthesis, and possesses an eyespot and two flagella with which it moves towards or away from light in order to optimize input of radiant energy. Motility contributes to C. reinhardtii fitness because it allows cells or colonies to achieve this optimum. Utilizing phototaxis to assay motility, we determined that newly evolved multicellular strains do not exhibit significant directional movement, even though the flagellae of their constituent unicells are present and active. In C. reinhardtii the first steps towards multicellularity in response to predation appear to result in a trade-off between motility and differential survivorship, a trade-off that must be overcome by further genetic change to ensure long-term success of the new multicellular organism.

The biosynthesis of nitrous oxide in the green alga Chlamydomonas reinhardtii.

Science.gov (United States)

Plouviez, Maxence; Wheeler, David; Shilton, Andy; Packer, Michael A; McLenachan, Patricia A; Sanz-Luque, Emanuel; Ocaña-Calahorro, Francisco; Fernández, Emilio; Guieysse, Benoit

2017-07-01

Over the last decades, several studies have reported emissions of nitrous oxide (N 2 O) from microalgal cultures and aquatic ecosystems characterized by a high level of algal activity (e.g. eutrophic lakes). As N 2 O is a potent greenhouse gas and an ozone-depleting pollutant, these findings suggest that large-scale cultivation of microalgae (and possibly, natural eutrophic ecosystems) could have a significant environmental impact. Using the model unicellular microalga Chlamydomonas reinhardtii, this study was conducted to investigate the molecular basis of microalgal N 2 O synthesis. We report that C. reinhardtii supplied with nitrite (NO 2 - ) under aerobic conditions can reduce NO 2 - into nitric oxide (NO) using either a mitochondrial cytochrome c oxidase (COX) or a dual enzymatic system of nitrate reductase (NR) and amidoxime-reducing component, and that NO is subsequently reduced into N 2 O by the enzyme NO reductase (NOR). Based on experimental evidence and published literature, we hypothesize that when nitrate (NO 3 - ) is the main Nitrogen source and the intracellular concentration of NO 2 - is low (i.e. under physiological conditions), microalgal N 2 O synthesis involves the reduction of NO 3 - to NO 2 - by NR followed by the reduction of NO 2 - to NO by the dual system involving NR. This microalgal N 2 O pathway has broad implications for environmental science and algal biology because the pathway of NO 3 - assimilation is conserved among microalgae, and because its regulation may involve NO. © 2017 The Authors The Plant Journal © 2017 John Wiley & Sons Ltd.

Site Energies of Active and Inactive Pheophytins in the Reaction Center of Photosystem II from Chlamydomonas Reinhardtii

Energy Technology Data Exchange (ETDEWEB)

Acharya, K.; Neupane, B.; Zazubovich, V.; Sayre, R. T.; Picorel, R.; Seibert, M.; Jankowiak, R.

2012-03-29

It is widely accepted that the primary electron acceptor in various Photosystem II (PSII) reaction center (RC) preparations is pheophytin {alpha} (Pheo {alpha}) within the D1 protein (Pheo{sub D1}), while Pheo{sub D2} (within the D2 protein) is photochemically inactive. The Pheo site energies, however, have remained elusive, due to inherent spectral congestion. While most researchers over the past two decades placed the Q{sub y}-states of Pheo{sub D1} and Pheo{sub D2} bands near 678-684 and 668-672 nm, respectively, recent modeling [Raszewski et al. Biophys. J. 2005, 88, 986-998; Cox et al. J. Phys. Chem. B 2009, 113, 12364-12374] of the electronic structure of the PSII RC reversed the assignment of the active and inactive Pheos, suggesting that the mean site energy of Pheo{sub D1} is near 672 nm, whereas Pheo{sub D2} ({approx}677.5 nm) and Chl{sub D1} ({approx}680 nm) have the lowest energies (i.e., the Pheo{sub D2}-dominated exciton is the lowest excited state). In contrast, chemical pigment exchange experiments on isolated RCs suggested that both pheophytins have their Q{sub y} absorption maxima at 676-680 nm [Germano et al. Biochemistry 2001, 40, 11472-11482; Germano et al. Biophys. J. 2004, 86, 1664-1672]. To provide more insight into the site energies of both Pheo{sub D1} and Pheo{sub D2} (including the corresponding Q{sub x} transitions, which are often claimed to be degenerate at 543 nm) and to attest that the above two assignments are most likely incorrect, we studied a large number of isolated RC preparations from spinach and wild-type Chlamydomonas reinhardtii (at different levels of intactness) as well as the Chlamydomonas reinhardtii mutant (D2-L209H), in which the active branch Pheo{sub D1} is genetically replaced with chlorophyll {alpha} (Chl {alpha}). We show that the Q{sub x}-/Q{sub y}-region site energies of Pheo{sub D1} and Pheo{sub D2} are {approx}545/680 nm and {approx}541.5/670 nm, respectively, in good agreement with our previous assignment

Antagonistic and synergistic effects of light irradiation on the effects of copper on Chlamydomonas reinhardtii

Energy Technology Data Exchange (ETDEWEB)

Cheloni, Giulia; Cosio, Claudia; Slaveykova, Vera I., E-mail: vera.slaveykova@unige.ch

2014-10-15

Highlights: • Light intensity and spectral composition affect Cu uptake and effects to C. reinhardtii. • High light (HL) reduced Cu effect on growth inhibition, oxidative stress and damage. • HL in combination with Cu up-regulated genes involved in the antioxidant responses. • HL with increased UVB radiation exacerbated Cu uptake and Cu-induced toxic effects. - Abstract: The present study showed the important role of light intensity and spectral composition on Cu uptake and effects on green alga Chlamydomonas reinhardtii. High-intenisty light (HL) increased cellular Cu concentrations, but mitigated the Cu-induced decrease in chlorophyll fluorescence, oxidative stress and lipid peroxidation at high Cu concentrations, indicating that Cu and HL interact in an antagonistic manner. HL up-regulated the transcription of genes involved in the antioxidant response in C. reinhardtii and thus reduced the oxidative stress upon exposure to Cu and HL. Combined exposure to Cu and UVBR resulted in an increase of cellular Cu contents and caused severe oxidative damage to the cells. The observed effects were higher than the sum of the effects corresponding to exposure to UVBR or Cu alone suggesting a synergistic interaction.

CDKL5 regulates flagellar length and localizes to the base of the flagella in Chlamydomonas

Science.gov (United States)

Tam, Lai-Wa; Ranum, Paul T.; Lefebvre, Paul A.

2013-01-01

The length of Chlamydomonas flagella is tightly regulated. Mutations in four genes—LF1, LF2, LF3, and LF4—cause cells to assemble flagella up to three times wild-type length. LF2 and LF4 encode protein kinases. Here we describe a new gene, LF5, in which null mutations cause cells to assemble flagella of excess length. The LF5 gene encodes a protein kinase very similar in sequence to the protein kinase CDKL5. In humans, mutations in this kinase cause a severe form of juvenile epilepsy. The LF5 protein localizes to a unique location: the proximal 1 μm of the flagella. The proximal localization of the LF5 protein is lost when genes that make up the proteins in the cytoplasmic length regulatory complex (LRC)—LF1, LF2, and LF3—are mutated. In these mutants LF5p becomes localized either at the distal tip of the flagella or along the flagellar length, indicating that length regulation involves, at least in part, control of LF5p localization by the LRC. PMID:23283985

Outlook in the application of Chlamydomonas reinhardtii chloroplast as a platform for recombinant protein production.

Science.gov (United States)

Shamriz, Shabnam; Ofoghi, Hamideh

Microalgae, also called microphytes, are a vast group of microscopic photosynthetic organisms living in aquatic ecosystems. Microalgae have attracted the attention of biotechnology industry as a platform for extracting natural products with high commercial value. During last decades, microalgae have been also used as cost-effective and easily scalable platform for the production of recombinant proteins with medical and industrial applications. Most progress in this field has been made with Chlamydomonas reinhardtii as a model organism mainly because of its simple life cycle, well-established genetics and ease of cultivation. However, due to the scarcity of existing infrastructure for commercial production and processing together with relatively low product yields, no recombinant products from C. reinhardtii have gained approval for commercial production and most of them are still in research and development. In this review, we focus on the chloroplast of C. reinhardtii as an algal recombinant expression platform and compare its advantages and disadvantages to other currently used expression systems. We then discuss the strategies for engineering the chloroplast of C. reinhardtii to produce recombinant cells and present a comprehensive overview of works that have used this platform for the expression of high-value products.

Adaptation prevents the extinction of Chlamydomonas reinhardtii under toxic beryllium

Directory of Open Access Journals (Sweden)

Beatriz Baselga-Cervera

2016-03-01

Full Text Available The current biodiversity crisis represents a historic challenge for natural communities: the environmental rate of change exceeds the population’s adaptation capability. Integrating both ecological and evolutionary responses is necessary to make reliable predictions regarding the loss of biodiversity. The race against extinction from an eco-evolutionary perspective is gaining importance in ecological risk assessment. Here, we performed a classical study of population dynamics—a fluctuation analysis—and evaluated the results from an adaption perspective. Fluctuation analysis, widely used with microorganisms, is an effective empirical procedure to study adaptation under strong selective pressure because it incorporates the factors that influence demographic, genetic and environmental changes. The adaptation of phytoplankton to beryllium (Be is of interest because human activities are increasing the concentration of Be in freshwater reserves; therefore, predicting the effects of human-induced pollutants is necessary for proper risk assessment. The fluctuation analysis was performed with phytoplankton, specifically, the freshwater microalgae Chlamydomonas reinhardtii, under acute Be exposure. High doses of Be led to massive microalgae death; however, by conducting a fluctuation analysis experiment, we found that C. reinhardtii was able to adapt to 33 mg/l of Be due to pre-existing genetic variability. The rescuing adapting genotype presented a mutation rate of 9.61 × 10−6 and a frequency of 10.42 resistant cells per million wild-type cells. The genetic adaptation pathway that was experimentally obtained agreed with the theoretical models of evolutionary rescue (ER. Furthermore, the rescuing genotype presented phenotypic and physiologic differences from the wild-type genotype, was 25% smaller than the Be-resistant genotype and presented a lower fitness and quantum yield performance. The abrupt distinctions between the wild-type and the Be

The diurnal logic of the expression of the chloroplast genome in Chlamydomonas reinhardtii.

Directory of Open Access Journals (Sweden)

Adam D Idoine

Full Text Available Chloroplasts are derived from cyanobacteria and have retained a bacterial-type genome and gene expression machinery. The chloroplast genome encodes many of the core components of the photosynthetic apparatus in the thylakoid membranes. To avoid photooxidative damage and production of harmful reactive oxygen species (ROS by incompletely assembled thylakoid protein complexes, chloroplast gene expression must be tightly regulated and co-ordinated with gene expression in the nucleus. Little is known about the control of chloroplast gene expression at the genome-wide level in response to internal rhythms and external cues. To obtain a comprehensive picture of organelle transcript levels in the unicellular model alga Chlamydomonas reinhardtii in diurnal conditions, a qRT-PCR platform was developed and used to quantify 68 chloroplast, 21 mitochondrial as well as 71 nuclear transcripts in cells grown in highly controlled 12 h light/12 h dark cycles. Interestingly, in anticipation of dusk, chloroplast transcripts from genes involved in transcription reached peak levels first, followed by transcripts from genes involved in translation, and finally photosynthesis gene transcripts. This pattern matches perfectly the theoretical demands of a cell "waking up" from the night. A similar trend was observed in the nuclear transcripts. These results suggest a striking internal logic in the expression of the chloroplast genome and a previously unappreciated complexity in the regulation of chloroplast genes.

Live cell imaging compatible immobilization of Chlamydomonas reinhardtii in microfluidic platform for biodiesel research.

Science.gov (United States)

Park, Jae Woo; Na, Sang Cheol; Nguyen, Thanh Qua; Paik, Sang-Min; Kang, Myeongwoo; Hong, Daewha; Choi, Insung S; Lee, Jae-Hyeok; Jeon, Noo Li

2015-03-01

This paper describes a novel surface immobilization method for live-cell imaging of Chlamydomonas reinhardtii for continuous monitoring of lipid droplet accumulation. Microfluidics allows high-throughput manipulation and analysis of single cells in precisely controlled microenvironment. Fluorescence imaging based quantitative measurement of lipid droplet accumulation in microalgae had been difficult due to their intrinsic motile behavior. We present a simple surface immobilization method using gelatin coating as the "biological glue." We take advantage of hydroxyproline (Hyp)-based non-covalent interaction between gelatin and the outer cell wall of microalgae to anchor the cells inside the microfluidic device. We have continuously monitored single microalgal cells for up to 6 days. The immobilized microalgae remain viable (viability was comparable to bulk suspension cultured controls). When exposed to wall shear stress, most of the cells remain attached up to 0.1 dyne/cm(2) . Surface immobilization allowed high-resolution, live-cell imaging of mitotic process in real time-which followed previously reported stages in mitosis of suspension cultured cells. Use of gelatin coated microfluidics devices can result in better methods for microalgae strain screening and culture condition optimization that will help microalgal biodiesel become more economically viable. © 2014 Wiley Periodicals, Inc.

Genome-wide identification of regulatory elements and reconstruction of gene regulatory networks of the green alga Chlamydomonas reinhardtii under carbon deprivation.

Directory of Open Access Journals (Sweden)

Flavia Vischi Winck

Full Text Available The unicellular green alga Chlamydomonas reinhardtii is a long-established model organism for studies on photosynthesis and carbon metabolism-related physiology. Under conditions of air-level carbon dioxide concentration [CO2], a carbon concentrating mechanism (CCM is induced to facilitate cellular carbon uptake. CCM increases the availability of carbon dioxide at the site of cellular carbon fixation. To improve our understanding of the transcriptional control of the CCM, we employed FAIRE-seq (formaldehyde-assisted Isolation of Regulatory Elements, followed by deep sequencing to determine nucleosome-depleted chromatin regions of algal cells subjected to carbon deprivation. Our FAIRE data recapitulated the positions of known regulatory elements in the promoter of the periplasmic carbonic anhydrase (Cah1 gene, which is upregulated during CCM induction, and revealed new candidate regulatory elements at a genome-wide scale. In addition, time series expression patterns of 130 transcription factor (TF and transcription regulator (TR genes were obtained for cells cultured under photoautotrophic condition and subjected to a shift from high to low [CO2]. Groups of co-expressed genes were identified and a putative directed gene-regulatory network underlying the CCM was reconstructed from the gene expression data using the recently developed IOTA (inner composition alignment method. Among the candidate regulatory genes, two members of the MYB-related TF family, Lcr1 (Low-CO 2 response regulator 1 and Lcr2 (Low-CO2 response regulator 2, may play an important role in down-regulating the expression of a particular set of TF and TR genes in response to low [CO2]. The results obtained provide new insights into the transcriptional control of the CCM and revealed more than 60 new candidate regulatory genes. Deep sequencing of nucleosome-depleted genomic regions indicated the presence of new, previously unknown regulatory elements in the C. reinhardtii genome

Flavodiiron Proteins Promote Fast and Transient O2 Photoreduction in Chlamydomonas.

Science.gov (United States)

Chaux, Frédéric; Burlacot, Adrien; Mekhalfi, Malika; Auroy, Pascaline; Blangy, Stéphanie; Richaud, Pierre; Peltier, Gilles

2017-07-01

During oxygenic photosynthesis, the reducing power generated by light energy conversion is mainly used to reduce carbon dioxide. In bacteria and archae, flavodiiron (Flv) proteins catalyze O 2 or NO reduction, thus protecting cells against oxidative or nitrosative stress. These proteins are found in cyanobacteria, mosses, and microalgae, but have been lost in angiosperms. Here, we used chlorophyll fluorescence and oxygen exchange measurement using [ 18 O]-labeled O 2 and a membrane inlet mass spectrometer to characterize Chlamydomonas reinhardtii flvB insertion mutants devoid of both FlvB and FlvA proteins. We show that Flv proteins are involved in a photo-dependent electron flow to oxygen, which drives most of the photosynthetic electron flow during the induction of photosynthesis. As a consequence, the chlorophyll fluorescence patterns are strongly affected in flvB mutants during a light transient, showing a lower PSII operating yield and a slower nonphotochemical quenching induction. Photoautotrophic growth of flvB mutants was indistinguishable from the wild type under constant light, but severely impaired under fluctuating light due to PSI photo damage. Remarkably, net photosynthesis of flv mutants was higher than in the wild type during the initial hour of a fluctuating light regime, but this advantage vanished under long-term exposure, and turned into PSI photo damage, thus explaining the marked growth retardation observed in these conditions. We conclude that the C. reinhardtii Flv participates in a Mehler-like reduction of O 2 , which drives a large part of the photosynthetic electron flow during a light transient and is thus critical for growth under fluctuating light regimes. © 2017 American Society of Plant Biologists. All Rights Reserved.

Thioredoxin-dependent Redox Regulation of Chloroplastic Phosphoglycerate Kinase from Chlamydomonas reinhardtii*

Science.gov (United States)

Morisse, Samuel; Michelet, Laure; Bedhomme, Mariette; Marchand, Christophe H.; Calvaresi, Matteo; Trost, Paolo; Fermani, Simona; Zaffagnini, Mirko; Lemaire, Stéphane D.

2014-01-01

In photosynthetic organisms, thioredoxin-dependent redox regulation is a well established mechanism involved in the control of a large number of cellular processes, including the Calvin-Benson cycle. Indeed, 4 of 11 enzymes of this cycle are activated in the light through dithiol/disulfide interchanges controlled by chloroplastic thioredoxin. Recently, several proteomics-based approaches suggested that not only four but all enzymes of the Calvin-Benson cycle may withstand redox regulation. Here, we characterized the redox features of the Calvin-Benson enzyme phosphoglycerate kinase (PGK1) from the eukaryotic green alga Chlamydomonas reinhardtii, and we show that C. reinhardtii PGK1 (CrPGK1) activity is inhibited by the formation of a single regulatory disulfide bond with a low midpoint redox potential (−335 mV at pH 7.9). CrPGK1 oxidation was found to affect the turnover number without altering the affinity for substrates, whereas the enzyme activation appeared to be specifically controlled by f-type thioredoxin. Using a combination of site-directed mutagenesis, thiol titration, mass spectrometry analyses, and three-dimensional modeling, the regulatory disulfide bond was shown to involve the not strictly conserved Cys227 and Cys361. Based on molecular mechanics calculation, the formation of the disulfide is proposed to impose structural constraints in the C-terminal domain of the enzyme that may lower its catalytic efficiency. It is therefore concluded that CrPGK1 might constitute an additional light-modulated Calvin-Benson cycle enzyme with a low activity in the dark and a TRX-dependent activation in the light. These results are also discussed from an evolutionary point of view. PMID:25202015

Calcium regulates ATP-sensitive microtubule binding by Chlamydomonas outer arm dynein.

Science.gov (United States)

Sakato, Miho; King, Stephen M

2003-10-31

The Chlamydomonas outer dynein arm contains three distinct heavy chains (alpha, beta, and gamma) that exhibit different motor properties. The LC4 protein, which binds 1-2 Ca2+ with KCa = 3 x 10-5 m, is associated with the gamma heavy chain and has been proposed to act as a sensor to regulate dynein motor function in response to alterations in intraflagellar Ca2+ levels. Here we genetically dissect the outer arm to yield subparticles containing different motor unit combinations and assess the microtubule-binding properties of these complexes both prior to and following preincubation with tubulin and ATP, which was used to inhibit ATP-insensitive (structural) microtubule binding. We observed that the alpha heavy chain exhibits a dominant Ca2+-independent ATP-sensitive MT binding activity in vitro that is inhibited by attachment of tubulin to the structural microtubule-binding domain. Furthermore, we show that ATP-sensitive microtubule binding by a dynein subparticle containing only the beta and gamma heavy chains does not occur at Ca2+ concentrations below pCa 6 but is maximally activated above pCa 5. This activity was not observed in mutant dyneins containing small deletions in the microtubule-binding region of the beta heavy chain or in dyneins that lack both the alpha heavy chain and the motor domain of the beta heavy chain. These findings strongly suggest that Ca2+ binding directly to a component of the dynein complex regulates ATP-sensitive interactions between the beta heavy chain and microtubules and lead to a model for how individual motor units are controlled within the outer dynein arm.

Crystallization and preliminary X-ray diffraction analysis of L,L-diaminopimelate aminotransferase (DapL) from Chlamydomonas reinhardtii.

Science.gov (United States)

Hudson, André O; Girón, Irma; Dobson, Renwick C J

2011-01-01

In the anabolic synthesis of diaminopimelate and lysine in plants and in some bacteria, the enzyme L,L-diaminopimelate aminotransferase (DapL; EC 2.6.1.83) catalyzes the conversion of tetrahydrodipicolinic acid (THDPA) to L,L-diaminopimelate, bypassing the DapD, DapC and DapE enzymatic steps in the bacterial acyl pathways. Here, the cloning, expression, purification, crystallization and preliminary X-ray diffraction analysis of DapL from the alga Chlamydomonas reinhardtii are presented. Protein crystals were grown in conditions containing 25% (w/v) PEG 3350 and 200 mM lithium sulfate and initially diffracted to ∼1.35 Å resolution. They belonged to space group P2(1)2(1)2(1), with unit-cell parameters a=58.9, b=91.8, c=162.9 Å. The data were processed to 1.55 Å resolution with an Rmerge of 0.081, an Rp.i.m. of 0.044, an Rr.i.m of 0.093 and a VM of 2.28 Å3 Da(-1).

L,L-diaminopimelate aminotransferase from Chlamydomonas reinhardtii: a target for algaecide development.

Science.gov (United States)

Dobson, Renwick C J; Girón, Irma; Hudson, André O

2011-01-01

In some bacterial species and photosynthetic cohorts, including algae, the enzyme L,L-diaminopimelate aminotransferase (DapL) (E.C. 2.6.1.83) is involved in the anabolism of the essential amino acid L-lysine. DapL catalyzes the conversion of tetrahydrodipicolinate (THDPA) to L,L-diaminopimelate (L,L-DAP), in one step bypassing the DapD, DapC and DapE enzymatic reactions present in the acyl DAP pathways. Here we present an in vivo and in vitro characterization of the DapL ortholog from the alga Chlamydomonas reinhardtii (Cr-DapL). The in vivo analysis illustrated that the enzyme is able to functionally complement the E. coli dap auxotrophs and was essential for plant development in Arabidopsis. In vitro, the enzyme was able to inter-convert THDPA and L,L-DAP, showing strong substrate specificity. Cr-DapL was dimeric in both solution and when crystallized. The structure of Cr-DapL was solved in its apo form, showing an overall architecture of a α/β protein with each monomer in the dimer adopting a pyridoxal phosphate-dependent transferase-like fold in a V-shaped conformation. The active site comprises residues from both monomers in the dimer and shows some rearrangement when compared to the apo-DapL structure from Arabidopsis. Since animals do not possess the enzymatic machinery necessary for the de novo synthesis of the amino acid L-lysine, enzymes involved in this pathway are attractive targets for the development of antibiotics, herbicides and algaecides.

L,L-diaminopimelate aminotransferase from Chlamydomonas reinhardtii: a target for algaecide development.

Directory of Open Access Journals (Sweden)

Renwick C J Dobson

Full Text Available In some bacterial species and photosynthetic cohorts, including algae, the enzyme L,L-diaminopimelate aminotransferase (DapL (E.C. 2.6.1.83 is involved in the anabolism of the essential amino acid L-lysine. DapL catalyzes the conversion of tetrahydrodipicolinate (THDPA to L,L-diaminopimelate (L,L-DAP, in one step bypassing the DapD, DapC and DapE enzymatic reactions present in the acyl DAP pathways. Here we present an in vivo and in vitro characterization of the DapL ortholog from the alga Chlamydomonas reinhardtii (Cr-DapL. The in vivo analysis illustrated that the enzyme is able to functionally complement the E. coli dap auxotrophs and was essential for plant development in Arabidopsis. In vitro, the enzyme was able to inter-convert THDPA and L,L-DAP, showing strong substrate specificity. Cr-DapL was dimeric in both solution and when crystallized. The structure of Cr-DapL was solved in its apo form, showing an overall architecture of a α/β protein with each monomer in the dimer adopting a pyridoxal phosphate-dependent transferase-like fold in a V-shaped conformation. The active site comprises residues from both monomers in the dimer and shows some rearrangement when compared to the apo-DapL structure from Arabidopsis. Since animals do not possess the enzymatic machinery necessary for the de novo synthesis of the amino acid L-lysine, enzymes involved in this pathway are attractive targets for the development of antibiotics, herbicides and algaecides.

Genetic analysis of suppressors of the PF10 mutation in Chlamydomonas reinhardtii

International Nuclear Information System (INIS)

Dutcher, S.K.; Gibbons, W.; Inwood, W.B.

1988-01-01

A mutation at the PF10 locus of the unicellular green alga Chlamydomonas reinhardtii leads to abnormal cell motility. The asymmetric form of the ciliary beat stroke characteristic of wild-type flagella is modified by this mutation to a nearly symmetric beat. We report here that this abnormal motility is a conditional phenotype that depends on light intensity. In the absence of light or under low light intensities, the motility is more severely impaired than at higher light intensities. By UV mutagenesis we obtained 11 intragenic and 70 extragenic strains that show reversion of the pf10 motility phenotype observed in low light. The intragenic events reverted the motility phenotype of the pf10 mutation completely. The extragenic events define at least seven suppressor loci; these map to linkage groups IV, VII, IX, XI, XII and XVII. Suppressor mutations at two of the seven loci (LIS1 and LIS2) require light for their suppressor activity. Forty-eight of the 70 extragenic suppressors were examined in heterozygous diploid cells; 47 of these mutants were recessive to the wild-type allele and one mutant (bop5-1) was dominant to the wild-type allele. Complementation analysis of the 47 recessive mutants showed unusual patterns. Most mutants within a recombinationally defined group failed to complement one another, although there were pairs that showed intra-allelic complementation. Additionally, some of the mutants at each recombinationally defined locus failed to complement mutants at other loci. They define dominant enhancers of one another

Light-harvesting complex gene expression is controlled by both transcriptional and post-transcriptional mechanisms during photoacclimation in Chlamydomonas reinhardtii

CERN Document Server

Durnford Dion, G; McKim, Sarah M; Sarchfield, Michelle L

2003-01-01

To compensate for increases in photon flux density (PFD), photosynthetic organisms possess mechanisms for reversibly modulating their photosynthetic apparatus to minimize photodamage. The photoacclimation response in Chlamydomonas reinhardtii was assessed following a 10-fold increase in PFD over 24h. In addition to a 50% reduction in the amount of chlorophyll and light-harvesting complexes (LHC) per cell, the expression of genes encoding polypeptides of the light-harvesting antenna were also affected. The abundance of Lhcb (a LHCH gene), Lhcb4 (a CP29-like gene), and Lhca (a LHCI gene) transcripts were reduced by 65 to 80%, within 1-2 h; however, the RNA levels of all three genes recovered to their low-light (LL) concentrations within 6-8 h. To determine the role of transcript turnover in this transient decline in abundance, the stability of all transcripts was measured. Although there was no change in the Lhcb or Lhca transcript turnover time, the Lhcb4 mRNA stability decreased 2.5-fold immediately following...

Toxicological effects of nanometer titanium dioxide (nano-TiO2) on Chlamydomonas reinhardtii.

Science.gov (United States)

Chen, Lanzhou; Zhou, Lina; Liu, Yongding; Deng, Songqiang; Wu, Hao; Wang, Gaohong

2012-10-01

The toxicological effects of nanometer titanium dioxide (nano-TiO2) on a unicellular green alga Chlamydomonas reinhardtii were assessed by investigating the changes of the physiology and cyto-ultrastructure of this species under treatment. We found that nano-TiO2 inhibited photosynthetic efficiency and cell growth, but the content of chlorophyll a content in algae did not change, while carotenoid and chlorophyll b contents increased. Malondialdehyde (MDA) content reached maximum values after 8h exposure and then decreased to a moderately low level at 72 h. Electron microscopy images indicated that as concentrations of nano-TiO2 increased, a large number of C. reinhardtii cells were noted to be damaged: the number of chloroplasts declined, various other organelles were degraded, plasmolysis occurred, and TiO2 nanoparticles were found to be located inside cell wall and membrane. It was also noted that cell surface was surrounded by TiO2 particles, which could present an obstacle to the exchange of substances between the cell and its surrounding environment. To sum up, the effect of nano-TiO2 on C. reinhardtii included cell surface aggregation, photosynthesis inhibition, lipid peroxidation and new protein synthesis, while the response of C. reinhardtii to nano-TiO2 was a rapid process which occurs during 24 h after exposing and may relate to physiological stress system to mitigate damage. Crown Copyright © 2012. Published by Elsevier Inc. All rights reserved.

Characterization of a Native Algae Species Chlamydomonas debaryana: Strain Selection, Bioremediation Ability, and Lipid Characterization

Directory of Open Access Journals (Sweden)

Bo Zhang

2014-08-01

Full Text Available Native microalgal species may offer a favorable combination of both wastewater treatment and biofuel production. In this research, a green microalgae, Chlamydomonas debaryana, was isolated from a local lagoon, screened for its lipid content using flow cytometry, and further identified with microscopic observations and DNA sequence analysis. When using swine wastewater as a medium, the biomass yields were between 0.6 and 1.62 g/L, giving a median value of 1.11 g/L. By increasing mass transfer rates and providing sufficient light intensity, the microalgal growth was intrinsically enhanced. The growth of C. debaryana reduced most nutritional contents of the wastewater except iron. When combining the microalgal growth and nutrient removal, C. debaryana was able to utilize 1.3 to 1.6×103 mg COD (chemical oxygen demand/g biomass, 55 to 90 ppm ammonia/g biomass, and 48 to 89 ppm phosphorous/g biomass, The lipid content of C. debaryana was 19.9 ± 4.3% of cell dry weight. The transesterified microalgal oil mostly consisted of 14 kinds of fatty acids, ranging from C5 to C22, which can be refined into renewable jet fuel or used as sources of omega-3 and omega-6 fatty acids.

Chlorophyll a is a favorable substrate for Chlamydomonas Mg-dechelatase encoded by STAY-GREEN.

Science.gov (United States)

Matsuda, Kaori; Shimoda, Yousuke; Tanaka, Ayumi; Ito, Hisashi

2016-12-01

Mg removal from chlorophyll by Mg-dechelatase is the first step of chlorophyll degradation. Recent studies showed that in Arabidopsis, Stay Green (SGR) encodes Mg-dechelatase. Though the Escherichia coli expression system is advantageous for investigating the properties of Mg-dechelatase, Arabidopsis Mg-dechelatase is not successfully expressed in E. coli. Chlamydomonas reinhardtii SGR (CrSGR) has a long, hydrophilic tail, suggesting that active CrSGR can be expressed in E. coli. After the incubation of chlorophyll a with CrSGR expressed in E. coli, pheophytin a accumulated, indicating that active CrSGR was expressed in E. coli. Substrate specificity of CrSGR against chlorophyll b and an intermediate molecule of the chlorophyll b degradation pathway was examined. CrSGR exhibited no activity against chlorophyll b and low activity against 7-hydroxymethyl chlorophyll a, consistent with the fact that chlorophyll b is degraded only after conversion to chlorophyll a. CrSGR exhibited low activity against divinyl chlorophyll a and chlorophyll a', and no activity against chlorophyllide a, protochlorophyll a, chlorophyll c 2 , and Zn-chlorophyll a. These observations indicate that chlorophyll a is the most favorable substrate for CrSGR. When CrSGR was expressed in Arabidopsis cells, the chlorophyll content decreased, further confirming that SGR has Mg-dechelating activity in chloroplasts. Copyright © 2016 Elsevier Masson SAS. All rights reserved.

Resistance to Phosphinothricin (Glufosinate) and Its Utilization as a Nitrogen Source by Chlamydomonas reinhardtii.

Science.gov (United States)

Franco, A R; Lopez-Siles, F J; Cardenas, J

1996-10-01

Wild-type strain 21gr of the green alga Chlamydomonas reinhardtii was resistant to the ammonium salt of l-phosphinothricin (PPT, also called glufosinate), an irreversible inhibitor of glutamine synthetase activity and the main active component of the herbicide BASTA (AgrEvo, Frankfurt am Main, Germany). Under the same conditions, however, this strain was highly sensitive to l-methionine-S-sulfoximine, a structural analog of PPT which has been reported to be 5 to 10 times less effective than PPT as an inhibitor in plants. Moreover, this alga was able to grow with PPT as the sole nitrogen source when this compound was provided at low concentrations. This utilization of PPT was dependent upon the addition of acetate and light and did not take place in the presence of ammonium. Resistance was due neither to the presence of N-acetyltransferase or transaminase activity nor to the presence of glutamine synthetase isoforms resistant to PPT. By using l-[methyl-(sup14)C]PPT, we demonstrated that resistance is due to lack of PPT transport into the cells. This strongly suggests that PPT and l-methionine-S-sulfoximine enter the cells through different systems. Growth with PPT is supported by its deamination by an l-amino acid oxidase activity which has been previously described to be located at the periplasm.

High-yield secretion of recombinant proteins from the microalga Chlamydomonas reinhardtii.

Science.gov (United States)

Ramos-Martinez, Erick Miguel; Fimognari, Lorenzo; Sakuragi, Yumiko

2017-09-01

Microalga-based biomanufacturing of recombinant proteins is attracting growing attention due to its advantages in safety, metabolic diversity, scalability and sustainability. Secretion of recombinant proteins can accelerate the use of microalgal platforms by allowing post-translational modifications and easy recovery of products from the culture media. However, currently, the yields of secreted recombinant proteins are low, which hampers the commercial application of this strategy. This study aimed at expanding the genetic tools for enhancing secretion of recombinant proteins in Chlamydomonas reinhardtii, a widely used green microalga as a model organism and a potential industrial biotechnology platform. We demonstrated that the putative signal sequence from C. reinhardtii gametolysin can assist the secretion of the yellow fluorescent protein Venus into the culture media. To increase the secretion yields, Venus was C-terminally fused with synthetic glycomodules comprised of tandem serine (Ser) and proline (Pro) repeats of 10 and 20 units [hereafter (SP) n , wherein n = 10 or 20]. The yields of the (SP) n -fused Venus were higher than Venus without the glycomodule by up to 12-fold, with the maximum yield of 15 mg/L. Moreover, the presence of the glycomodules conferred an enhanced proteolytic protein stability. The Venus-(SP) n proteins were shown to be glycosylated, and a treatment of the cells with brefeldin A led to a suggestion that glycosylation of the (SP) n glycomodules starts in the endoplasmic reticulum (ER). Taken together, the results demonstrate the utility of the gametolysin signal sequence and (SP) n glycomodule to promote a more efficient biomanufacturing of microalgae-based recombinant proteins. © 2017 The Authors. Plant Biotechnology Journal published by Society for Experimental Biology and The Association of Applied Biologists and John Wiley & Sons Ltd.

Experimental Definition and Validation of Protein Coding Transcripts in Chlamydomonas reinhardtii

Energy Technology Data Exchange (ETDEWEB)

Kourosh Salehi-Ashtiani; Jason A. Papin

2012-01-13

Algal fuel sources promise unsurpassed yields in a carbon neutral manner that minimizes resource competition between agriculture and fuel crops. Many challenges must be addressed before algal biofuels can be accepted as a component of the fossil fuel replacement strategy. One significant challenge is that the cost of algal fuel production must become competitive with existing fuel alternatives. Algal biofuel production presents the opportunity to fine-tune microbial metabolic machinery for an optimal blend of biomass constituents and desired fuel molecules. Genome-scale model-driven algal metabolic design promises to facilitate both goals by directing the utilization of metabolites in the complex, interconnected metabolic networks to optimize production of the compounds of interest. Using Chlamydomonas reinhardtii as a model, we developed a systems-level methodology bridging metabolic network reconstruction with annotation and experimental verification of enzyme encoding open reading frames. We reconstructed a genome-scale metabolic network for this alga and devised a novel light-modeling approach that enables quantitative growth prediction for a given light source, resolving wavelength and photon flux. We experimentally verified transcripts accounted for in the network and physiologically validated model function through simulation and generation of new experimental growth data, providing high confidence in network contents and predictive applications. The network offers insight into algal metabolism and potential for genetic engineering and efficient light source design, a pioneering resource for studying light-driven metabolism and quantitative systems biology. Our approach to generate a predictive metabolic model integrated with cloned open reading frames, provides a cost-effective platform to generate metabolic engineering resources. While the generated resources are specific to algal systems, the approach that we have developed is not specific to algae and

Comparison of CO(2) and bicarbonate as inorganic carbon sources for triacylglycerol and starch accumulation in Chlamydomonas reinhardtii.

Science.gov (United States)

Gardner, Robert D; Lohman, Egan; Gerlach, Robin; Cooksey, Keith E; Peyton, Brent M

2013-01-01

Microalgae are capable of accumulating high levels of lipids and starch as carbon storage compounds. Investigation into the metabolic activities involved in the synthesis of these compounds has escalated since these compounds can be used as precursors for food and fuel. Here, we detail the results of a comprehensive analysis of Chlamydomonas reinhardtii using high or low inorganic carbon concentrations and speciation between carbon dioxide and bicarbonate, and the effects these have on inducing lipid and starch accumulation during nitrogen depletion. High concentrations of CO(2) (5%; v/v) produced the highest amount of biofuel precursors, transesterified to fatty acid methyl esters, but exhibited rapid accumulation and degradation characteristics. Low CO(2) (0.04%; v/v) caused carbon limitation and minimized triacylglycerol (TAG) and starch accumulation. High bicarbonate caused a cessation of cell cycling and accumulation of both TAG and starch that was more stable than the other experimental conditions. Starch accumulated prior to TAG and then degraded as maximum TAG was reached. This suggests carbon reallocation from starch-based to TAG-based carbon storage. Copyright © 2012 Wiley Periodicals, Inc.

Biodegradation of carbamazepine using freshwater microalgae Chlamydomonas mexicana and Scenedesmus obliquus and the determination of its metabolic fate.

Science.gov (United States)

Xiong, Jiu-Qiang; Kurade, Mayur B; Abou-Shanab, Reda A I; Ji, Min-Kyu; Choi, Jaeyoung; Kim, Jong Oh; Jeon, Byong-Hun

2016-04-01

This study evaluated the toxicity and cellular stresses of carbamazepine (CBZ) on Chlamydomonas mexicana and Scenedesmus obliquus, and its biodegradation by both microalgal species. The growth of both microalgal species decreased with increase of CBZ concentration. The growth of S. obliquus was significantly inhibited (97%) at 200 mg CBZ L(-1), as compared to the control after 10days; whereas, C. mexicana showed 30% inhibition at the same experimental conditions. Biochemical characteristics including total chlorophyll, carotenoid contents and enzyme activities (SOD and CAT) for both species were affected by CBZ at relatively high concentration. C. mexicana and S. obliquus could achieve a maximum of 35% and 28% biodegradation of CBZ, respectively. Two metabolites (10,11-dihydro-10,11-expoxycarbamazepine and n-hydroxy-CBZ) were identified by UPLC-MS, as a result of CBZ biodegradation by C. mexicana. This study demonstrated that C. mexicana was more tolerant to CBZ and could be used for treatment of CBZ contaminated wastewater. Copyright © 2016 Elsevier Ltd. All rights reserved.

Bending patterns of chlamydomonas flagella: III. A radial spoke head deficient mutant and a central pair deficient mutant.

Science.gov (United States)

Brokaw, C J; Luck, D J

1985-01-01

Flash photomicrography at frequencies up to 300 Hz and computer-assisted image analysis have been used to obtain parameters describing the flagellar bending patterns of mutants of Chlamydomonas reinhardtii. All strains contained the uni1 mutation, to facilitate photography. The radial spoke head deficient mutant pf17, and the central pair deficient mutant, pf15, in combination with suppressor mutations that restore motility without restoring the ultrastructural or biochemical deficiencies, both generate forward mode bending patterns with increased shear amplitude and decreased asymmetry relative to the "wild-type" uni1 flagella described previously. In the reverse beating mode, the suppressed pf17 mutants generate reverse bending patterns with large shear amplitudes. Reverse beating of the suppressed pf15 mutants is rare. There is a reciprocal relationship between increased shear amplitude and decreased beat frequency, so that the velocity of sliding between flagellar microtubules is not increased by an increase in shear amplitude. The suppressor mutations alone cause decreased frequency and sliding velocity in both forward and reverse mode beating, with little change in shear amplitude or symmetry.

[The impact of melafen on the expression of chloroplastic chaperone protein HSP70B and photosynthetic pigments in cells of Chlamydomonas reinhardtii].

Science.gov (United States)

Ermokhina, O V; Belkina, G G; Oleskina, Iu P; Fattakhov, S G; Iurina, N P

2009-01-01

The effects of growth regulator of the new generation-melamine salt of bis(oxymethyl)phosphine acid (melafen)--on culture growth, pigment and protein content, and the induction of protective chloroplastic chaperone HSP70B in Chlamydomonas reinhardtii CW15 cells were studied. Melafen exhibited 10-30% growth inhibition at 10(-9)-10(-2)% concentration. At 10(-9)-10(-4)% of melafen electrophoretic concentration, the pattern of cellular proteins was similar to the control. The alterations in protein content of algae cells were detected only at 10(-2)% concentration. The content of chlorophyll and carotenoids in melafen-treated cells was 17-40% lower than in the control. Melafen at 10(-9)-109-2)% concentration inhibited HSP70B induction by 39-43% compared to untreated cells. The potential mechanism of melafen effect might involve its influence on nuclear gene expression.

Gradual plasticity alters population dynamics in variable environments: thermal acclimation in the green alga Chlamydomonas reinhartdii.

Science.gov (United States)

Kremer, Colin T; Fey, Samuel B; Arellano, Aldo A; Vasseur, David A

2018-01-10

Environmental variability is ubiquitous, but its effects on populations are not fully understood or predictable. Recent attention has focused on how rapid evolution can impact ecological dynamics via adaptive trait change. However, the impact of trait change arising from plastic responses has received less attention, and is often assumed to optimize performance and unfold on a separate, faster timescale than ecological dynamics. Challenging these assumptions, we propose that gradual plasticity is important for ecological dynamics, and present a study of the plastic responses of the freshwater green algae Chlamydomonas reinhardtii as it acclimates to temperature changes. First, we show that C. reinhardtii 's gradual acclimation responses can both enhance and suppress its performance after a perturbation, depending on its prior thermal history. Second, we demonstrate that where conventional approaches fail to predict the population dynamics of C. reinhardtii exposed to temperature fluctuations, a new model of gradual acclimation succeeds. Finally, using high-resolution data, we show that phytoplankton in lake ecosystems can experience thermal variation sufficient to make acclimation relevant. These results challenge prevailing assumptions about plasticity's interactions with ecological dynamics. Amidst the current emphasis on rapid evolution, it is critical that we also develop predictive methods accounting for plasticity. © 2018 The Author(s).

Phytotoxicity of 15 common pharmaceuticals on the germination of Lactuca sativa and photosynthesis of Chlamydomonas reinhardtii.

Science.gov (United States)

Pino, Ma Rosa; Muñiz, Selene; Val, Jonatan; Navarro, Enrique

2016-11-01

Pharmaceuticals reach terrestrial environments through the application of treated wastewaters and biosolids to agricultural soils. We have investigated the toxicity of 15 common pharmaceuticals, classified as nonsteroidal anti-inflammatory drugs (NSAIDs), blood lipid-lowering agents, β-blockers and antibiotics, in two photosynthetic organisms. Twelve pharmaceuticals caused inhibitory effects on the radicle and hypocotyl elongation of Lactuca sativa seeds. The EC 50 values obtained were in the range of 170-5656 mg L -1 in the case of the radicle and 188-4558 mg L -1 for the hypocotyl. Propranolol was the most toxic drug for both root and hypocotyl elongation, followed by the NSAIDs, then gemfibrozil and tetracycline. Other effects, such as root necrosis, inhibition of root growth and curly hairs, were detected. However, even at the highest concentrations tested (3000 mg L -1 ), seed germination was not affected. NSAIDs decreased the photosynthetic yield of Chlamydomonas reinhardtii, but only salicylic acid showed EC 50 values below 1000 mg L -1 . The first effects detected at low concentrations, together with the concentrations found in environmental samples, indicate that the use of biosolids and wastewaters containing pharmaceuticals should be regulated and their compositions assessed in order to prevent medium- and long-term impacts on agricultural soils and crops.

Application of proton exchange membrane fuel cells for the monitoring and direct usage of biohydrogen produced by Chlamydomonas reinhardtii

Energy Technology Data Exchange (ETDEWEB)

Oncel, S.; Vardar-Sukan, F. [Department of Bioengineering, Faculty of Engineering, Ege University, 35100 Bornova, Izmir (Turkey)

2011-01-01

Photo-biologically produced hydrogen by Chlamydomonas reinhardtii is integrated with a proton exchange (PEM) fuel cell for online electricity generation. To investigate the fuel cell efficiency, the effect of hydrogen production on the open circuit fuel cell voltage is monitored during 27 days of batch culture. Values of volumetric hydrogen production, monitored by the help of the calibrated water columns, are related with the open circuit voltage changes of the fuel cell. From the analysis of this relation a dead end configuration is selected to use the fuel cell in its best potential. After the open circuit experiments external loads are tested for their effects on the fuel cell voltage and current generation. According to the results two external loads are selected for the direct usage of the fuel cell incorporating with the photobioreactors (PBR). Experiments with the PEM fuel cell generate a current density of 1.81 mA cm{sup -2} for about 50 h with 10 {omega} load and 0.23 mA cm{sup -2} for about 80 h with 100 {omega} load. (author)

Effect of mutagen combined action on Chlamydomonas reinhardtii cells. I. Lethal effect dependence on the sequence of mutagen application and on cultivation conditions

Energy Technology Data Exchange (ETDEWEB)

Vlcek, D; Podstavkova, S; Dubovsky, J [Komenskeho Univ., Bratislava (Czechoslovakia). Prirodovedecka Fakulta

1978-01-01

The effect was investigated of single and combined actions of alkylnitrosourea derivatives (N-methyl-N-nitrosourea and N-ethyl-N-nitrosourea) and UV-radiation on the survival of cells of Chlamydomonas reinhardtii algae in dependence on the sequence of application of mutagens and on the given conditions of cultivation following mutagen activity. In particular, the single phases were investigated of the total lethal effect, i.e., the death of cells before division and their death after division. The most pronounced changes in dependence on the sequence of application of mutagens and on the given conditions of cultivation were noted in cell death before division. In dependence on the sequence of application of mutagens, the effect of the combined action on the survival of cells changed from an additive (alkylnitrosourea + UV-radiation) to a protective effect (UV-radiation + alkylnitrosourea).

Deletion of Proton Gradient Regulation 5 (PGR5) and PGR5-Like 1 (PGRL1) proteins promote sustainable light-driven hydrogen production in Chlamydomonas reinhardtii due to increased PSII activity under sulfur deprivation.

Science.gov (United States)

Steinbeck, Janina; Nikolova, Denitsa; Weingarten, Robert; Johnson, Xenie; Richaud, Pierre; Peltier, Gilles; Hermann, Marita; Magneschi, Leonardo; Hippler, Michael

2015-01-01

Continuous hydrogen photo-production under sulfur deprivation was studied in the Chlamydomonas reinhardtii pgr5 pgrl1 double mutant and respective single mutants. Under medium light conditions, the pgr5 exhibited the highest performance and produced about eight times more hydrogen than the wild type, making pgr5 one of the most efficient hydrogen producer reported so far. The pgr5 pgrl1 double mutant showed an increased hydrogen burst at the beginning of sulfur deprivation under high light conditions, but in this case the overall amount of hydrogen produced by pgr5 pgrl1 as well as pgr5 was diminished due to photo-inhibition and increased degradation of PSI. In contrast, the pgrl1 was effective in hydrogen production in both high and low light. Blocking photosynthetic electron transfer by DCMU stopped hydrogen production almost completely in the mutant strains, indicating that the main pathway of electrons toward enhanced hydrogen production is via linear electron transport. Indeed, PSII remained more active and stable in the pgr mutant strains as compared to the wild type. Since transition to anaerobiosis was faster and could be maintained due to an increased oxygen consumption capacity, this likely preserves PSII from photo-oxidative damage in the pgr mutants. Hence, we conclude that increased hydrogen production under sulfur deprivation in the pgr5 and pgrl1 mutants is caused by an increased stability of PSII permitting sustainable light-driven hydrogen production in Chlamydomonas reinhardtii.

Chlamydomonas reinhardtii responding to high light: a role for 2-propenal (acrolein).

Science.gov (United States)

Roach, Thomas; Baur, Theresa; Stöggl, Wolfgang; Krieger-Liszkay, Anja

2017-09-01

High light causes photosystem II to generate singlet oxygen ( 1 O 2 ), a reactive oxygen species (ROS) that can react with membrane lipids, releasing reactive electrophile species (RES), such as acrolein. To investigate how RES may contribute to light stress responses, Chlamydomonas reinhardtii was high light-treated in photoautotrophic and mixotrophic conditions and also in an oxygen-enriched atmosphere to elevate ROS production. The responses were compared to exogenous acrolein. Non-photochemical quenching (NPQ) was higher in photoautotrophic cells, as a consequence of a more de-epoxidized state of the xanthophyll cycle pool and more LHCSR3 protein, showing that photosynthesis was under more pressure than in mixotrophic cells. Photoautotrophic cells had lowered α-tocopherol and β-carotene contents and a higher level of protein carbonylation, indicators of elevated 1 O 2 production. Levels of glutathione, glutathione peroxidase (GPX5) and glutathione-S-transferase (GST1), important antioxidants against RES, were also increased in photoautotrophic cells. In parallel to the wild-type, the LHCSR3-deficient npq4 mutant was high light-treated, which in photoautotrophic conditions exhibited particular sensitivity under elevated oxygen, the treatment that induced the highest RES levels, including acrolein. The npq4 mutant had more GPX5 and GST1 alongside higher levels of carbonylated protein and a more oxidized glutathione redox state. In wild-type cells glutathione contents doubled after 4 h treatment, either with high light under elevated oxygen or with a non-critical dose (600 ppm) of acrolein. Exogenous acrolein also increased GST1 levels, but not GPX5. Overall, RES-associated oxidative damage and glutathione metabolism are prominently associated with light stress and potentially in signaling responses of C. reinhardtii. © 2017 Scandinavian Plant Physiology Society.

LHCSR1 induces a fast and reversible pH-dependent fluorescence quenching in LHCII in Chlamydomonas reinhardtii cells.

Science.gov (United States)

Dinc, Emine; Tian, Lijin; Roy, Laura M; Roth, Robyn; Goodenough, Ursula; Croce, Roberta

2016-07-05

To avoid photodamage, photosynthetic organisms are able to thermally dissipate the energy absorbed in excess in a process known as nonphotochemical quenching (NPQ). Although NPQ has been studied extensively, the major players and the mechanism of quenching remain debated. This is a result of the difficulty in extracting molecular information from in vivo experiments and the absence of a validation system for in vitro experiments. Here, we have created a minimal cell of the green alga Chlamydomonas reinhardtii that is able to undergo NPQ. We show that LHCII, the main light harvesting complex of algae, cannot switch to a quenched conformation in response to pH changes by itself. Instead, a small amount of the protein LHCSR1 (light-harvesting complex stress related 1) is able to induce a large, fast, and reversible pH-dependent quenching in an LHCII-containing membrane. These results strongly suggest that LHCSR1 acts as pH sensor and that it modulates the excited state lifetimes of a large array of LHCII, also explaining the NPQ observed in the LHCSR3-less mutant. The possible quenching mechanisms are discussed.

A mutation in the centriole-associated protein centrin causes genomic instability via increased chromosome loss in Chlamydomonas reinhardtii

Directory of Open Access Journals (Sweden)

Marshall Wallace F

2005-05-01

Full Text Available Abstract Background The role of centrioles in mitotic spindle function remains unclear. One approach to investigate mitotic centriole function is to ask whether mutation of centriole-associated proteins can cause genomic instability. Results We addressed the role of the centriole-associated EF-hand protein centrin in genomic stability using a Chlamydomonas reinhardtii centrin mutant that forms acentriolar bipolar spindles and lacks the centrin-based rhizoplast structures that join centrioles to the nucleus. Using a genetic assay for loss of heterozygosity, we found that this centrin mutant showed increased genomic instability compared to wild-type cells, and we determined that the increase in genomic instability was due to a 100-fold increase in chromosome loss rates compared to wild type. Live cell imaging reveals an increased rate in cell death during G1 in haploid cells that is consistent with an elevated rate of chromosome loss, and analysis of cell death versus centriole copy number argues against a role for multipolar spindles in this process. Conclusion The increased chromosome loss rates observed in a centrin mutant that forms acentriolar spindles suggests a role for centrin protein, and possibly centrioles, in mitotic fidelity.

Hydrogen production by Chlamydomonas reinhardtii: an elaborate interplay of electron sources and sinks

International Nuclear Information System (INIS)

Hemschemeier, A; Happe, T.; Fouchard, S; Cournac, L; Peltier, G.

2008-01-01

The unicellular green alga Chlamydomonas reinhardtii possesses a [FeFe]-hydrogenase HydA1 (EC 1.12.7.2), which is coupled to the photosynthetic electron transport chain. Large amounts of H 2 are produced in a light-dependent reaction for several days when C. reinhardtii cells are deprived of sulfur. Under these conditions, the cells drastically change their physiology from aerobic photosynthetic growth to an anaerobic resting state. The understanding of the underlying physiological processes is not only important for getting further insights into the adaptability of photosynthesis, but will help to optimize the biotechnological application of algae as H 2 producers. Two of the still most disputed questions regarding H 2 generation by C. reinhardtii concern the electron source for H 2 evolution and the competition of the hydrogenase with alternative electron sinks. We analyzed the H 2 metabolism of S-depleted C. reinhardtii cultures utilizing a special mass spectrometer setup and investigated the influence of photosystem II (PSII)- or ribulose-bisphosphate-carboxylase/oxygenase (Rubisco)-deficiency. We show that electrons for H 2 -production are provided both by PSII activity and by a non-photochemical plastoquinone reduction pathway, which is dependent on previous PSII activity. In a Rubisco-deficient strain, which produces H 2 also in the presence of sulfur, H 2 generation seems to be the only significant electron sink for PSII activity and rescues this strain at least partially from a light-sensitive phenotype.The latter indicates that the down-regulation of assimilatory pathways in S-deprived C. reinhardtii cells is one of the important prerequisites for a sustained H 2 evolution. (authors)

Valorization of Spent Escherichia coli Media Using Green Microalgae Chlamydomonas reinhardtii and Feedstock Production

Directory of Open Access Journals (Sweden)

Jian-Guo Zhang

2017-06-01

Full Text Available The coupling of Chlamydomonas reinhardtii biomass production for nutrients removal of Escherichia coli anaerobic broth (EAB is thought to be an economically feasible option for the cultivation of microalgae. The feasibility of growing microalgae in using EAB high in nutrients for the production of more biomass was examined. EAB comprised of nutrient-abundant effluents, which can be used to produce microalgae biomass and remove environment pollutant simultaneously. In this study, C. reinhardtii 21gr (cc1690 was cultivated in different diluted E. coli anaerobic broth supplemented with trace elements under mixotrophic and heterotrophic conditions. The results showed that C. reinhardtii grown in 1×, 1/2×, 1/5× and 1/10×E. coli anaerobic broth under mixotrophic conditions exhibited specific growth rates of 2.71, 2.68, 1.45, and 1.13 day-1, and biomass production of 201.9, 184.2, 175.5, and 163.8 mg L-1, respectively. Under heterotrophic conditions, the specific growth rates were 1.80, 1.86, 1.75, and 1.02 day-1, and biomass production were 45.6, 29.4, 15.8, and 12.1 mg L-1, respectively. The removal efficiency of chemical oxygen demand, total-nitrogen and total-phosphorus from 1×E. coli anaerobic broth was 21.51, 22.41, and 15.53%. Moreover, the dry biomass had relatively high carbohydrate (44.3% and lipid content (18.7%. Therefore, this study provides an environmentally sustainable as well economical method for biomass production in promising model microalgae and subsequently paves the way for industrial use.

Not changes in membrane fluidity but proteotoxic stress triggers heat shock protein expression in Chlamydomonas reinhardtii.

Science.gov (United States)

Rütgers, Mark; Muranaka, Ligia Segatto; Schulz-Raffelt, Miriam; Thoms, Sylvia; Schurig, Juliane; Willmund, Felix; Schroda, Michael

2017-12-01

A conserved reaction of all organisms exposed to heat stress is an increased expression of heat shock proteins (HSPs). Several studies have proposed that HSP expression in heat-stressed plant cells is triggered by an increased fluidity of the plasma membrane. Among the main lines of evidence in support of this model are as follows: (a) the degree of membrane lipid saturation was higher in cells grown at elevated temperatures and correlated with a lower amplitude of HSP expression upon a temperature upshift, (b) membrane fluidizers induce HSP expression at physiological temperatures, and (c) membrane rigidifier dimethylsulfoxide dampens heat-induced HSP expression. Here, we tested whether this holds also for Chlamydomonas reinhardtii. We show that heat-induced HSP expression in cells grown at elevated temperatures was reduced because they already contained elevated levels of cytosolic HSP70A/90A that apparently act as negative regulators of heat shock factor 1. We find that membrane rigidifier dimethylsulfoxide impaired translation under heat stress conditions and that membrane fluidizer benzyl alcohol not only induced HSP expression but also caused protein aggregation. These findings support the classical model for the cytosolic unfolded protein response, according to which HSP expression is induced by the accumulation of unfolded proteins. Hence, the membrane fluidity model should be reconsidered. © 2017 John Wiley & Sons Ltd.

Bioavailability of wastewater derived dissolved organic nitrogen to green microalgae Selenastrum capricornutum, Chlamydomonas reinhardtii, and Chlorella vulgaris with/without presence of bacteria.

Science.gov (United States)

Sun, Jingyi; Simsek, Halis

2017-07-01

Effluent dissolved organic nitrogen (DON) is problematic in nutrient sensitive surface waters and needs to be reduced to meet demanding total dissolved nitrogen discharge limits. Bioavailable DON (ABDON) is a portion of DON utilized by algae or algae+bacteria, while biodegradable DON (BDON) is a portion of DON decomposable by bacteria. ABDON and BDON in a two-stage trickling filter (TF) wastewater treatment plant was evaluated using three different microalgal species, Selenastrum capricornutum, Chlamydomonas reinhardtii and Chlorella vulgaris and mixed cultured bacteria. Results showed that up to 80% of DON was bioavailable to algae or algae+bacteria inoculum while up to 60% of DON was biodegradable in all the samples. Results showed that C. reinhardtii and C. vulgaris can be used as a test species the same as S. capricornutum since there were no significant differences among these three algae species based on their ability to remove nitrogen species. Copyright © 2017. Published by Elsevier B.V.

Identification and molecular characterization of the second Chlamydomonas gun4 mutant, gun4-II [v2; ref status: indexed, http://f1000r.es/1id

Directory of Open Access Journals (Sweden)

Phillip B Grovenstein

2013-07-01

Full Text Available The green micro-alga Chlamydomonas reinhardtii is an elegant model organism to study oxygenic photosynthesis. Chlorophyll (Chl and heme are major tetrapyrroles that play an essential role in photosynthesis and respiration. These tetrapyrroles are synthesized via a common branched pathway that involves mainly enzymes, encoded by nuclear genes. One of the enzymes in the pathway is Mg chelatase (MgChel. MgChel catalyzes insertion of Mg2+ into protoporphyrin IX (PPIX, proto to form Magnesium-protoporphyrin IX (MgPPIX, Mgproto, the first biosynthetic intermediate in the Chl branch. The GUN4 (genomes uncoupled 4 protein is not essential for the MgChel activity but has been shown to significantly stimulate its activity. We have isolated a light sensitive mutant, 6F14, by random DNA insertional mutagenesis. 6F14 cannot tolerate light intensities higher than 90-100 μmol photons m-2 s-1. It shows a light intensity dependent progressive photo-bleaching. 6F14 is incapable of photo-autotrophic growth under light intensity higher than 100 μmol photons m-2 s-1. PCR based analyses show that in 6F14 the insertion of the plasmid outside the GUN4 locus has resulted in a genetic rearrangement of the GUN4 gene and possible deletions in the genomic region flanking the GUN4 gene. Our gun4 mutant has a Chl content very similar to that in the wild type in the dark and is very sensitive to fluctuations in the light intensity in the environment unlike the earlier identified Chlamydomonas gun4 mutant. Complementation with a functional copy of the GUN4 gene restored light tolerance, Chl biosynthesis and photo-autotrophic growth under high light intensities in 6F14. 6F14 is the second gun4 mutant to be identified in C. reinhardtii. Additionally, we show that our two gun4 complements over-express the GUN4 protein and show a higher Chl content per cell compared to that in the wild type strain.

Functional specificity of cardiolipin synthase revealed by the identification of a cardiolipin synthase CrCLS1 in Chlamydomonas reinhardtii

Directory of Open Access Journals (Sweden)

Chun-Hsien eHung

2016-01-01

Full Text Available Phosphatidylglycerol (PG and cardiolipin (CL are two essential classes of phospholipid in plants and algae. Phosphatidylglycerophosphate synthase (PGPS and cardiolipin synthase (CLS involved in the biosynthesis of PG and CL belong to CDP-alcohol phosphotransferase and share overall amino acid sequence homology. However, it remains elusive whether PGPS and CLS are functionally distinct in vivo. Here, we report identification of a gene encoding CLS in Chlamydomonas reinhardtii, CrCLS1, and its functional compatibility. Whereas CrCLS1 did not complement the growth phenotype of a PGPS mutant of Synechocystis sp. PCC 6803, it rescued the temperature-sensitive growth phenotype, growth profile with different carbon sources, phospholipid composition and enzyme activity of ∆crd1, a CLS mutant of Saccharomyces cerevisiae. These results suggest that CrCLS1 encodes a functional CLS of C. reinhardtii as the first identified algal CLS, whose enzyme function is distinct from that of PGPSs from C. reinhardtii. Comparison of CDP-alcohol phosphotransferase motif between PGPS and CLS among different species revealed a possible additional motif that might define the substrate specificity of these closely related enzymes.

Cellular oxido-reductive proteins of Chlamydomonas reinhardtii control the biosynthesis of silver nanoparticles

Directory of Open Access Journals (Sweden)

Barwal Indu

2011-12-01

Full Text Available Abstract Background Elucidation of molecular mechanism of silver nanoparticles (SNPs biosynthesis is important to control its size, shape and monodispersity. The evaluation of molecular mechanism of biosynthesis of SNPs is of prime importance for the commercialization and methodology development for controlling the shape and size (uniform distribution of SNPs. The unicellular algae Chlamydomonas reinhardtii was exploited as a model system to elucidate the role of cellular proteins in SNPs biosynthesis. Results The C. reinhardtii cell free extract (in vitro and in vivo cells mediated synthesis of silver nanoparticles reveals SNPs of size range 5 ± 1 to 15 ± 2 nm and 5 ± 1 to 35 ± 5 nm respectively. In vivo biosynthesized SNPs were localized in the peripheral cytoplasm and at one side of flagella root, the site of pathway of ATP transport and its synthesis related enzymes. This provides an evidence for the involvement of oxidoreductive proteins in biosynthesis and stabilization of SNPs. Alteration in size distribution and decrease of synthesis rate of SNPs in protein-depleted fractions confirmed the involvement of cellular proteins in SNPs biosynthesis. Spectroscopic and SDS-PAGE analysis indicate the association of various proteins on C. reinhardtii mediated in vivo and in vitro biosynthesized SNPs. We have identified various cellular proteins associated with biosynthesized (in vivo and in vitro SNPs by using MALDI-MS-MS, like ATP synthase, superoxide dismutase, carbonic anhydrase, ferredoxin-NADP+ reductase, histone etc. However, these proteins were not associated on the incubation of pre-synthesized silver nanoparticles in vitro. Conclusion Present study provides the indication of involvement of molecular machinery and various cellular proteins in the biosynthesis of silver nanoparticles. In this report, the study is mainly focused towards understanding the role of diverse cellular protein in the synthesis and capping of silver

The mechanism of anthracene interaction with photosynthetic apparatus: A study using intact cells, thylakoid membranes and PS II complexes isolated from Chlamydomonas reinhardtii

International Nuclear Information System (INIS)

Aksmann, Anna; Shutova, Tatiana; Samuelsson, Goeran; Tukaj, Zbigniew

2011-01-01

Intact cells of Chlamydomonas reinhardtii as well as isolated thylakoid membranes and photosystem II complexes were used to examine a possible mechanism of anthracene (ANT) interaction with the photosynthetic apparatus. Since ANT concentrations above 1 mM were required to significantly inhibit the rate of oxygen evolution in PS II membrane fragments it may indicate that the toxicant did not directly interact with this photosystem. On the other hand, stimulation of oxygen uptake by ANT-treated thylakoids suggested that ANT could either act as an artificial electron acceptor in the photosynthetic electron transport chain or function as an uncoupler. Electron transfer from excited chlorophyll to ANT is impossible due to the very low reduction potential of ANT and therefore we propose that toxic concentrations of ANT increase the thylakoid membrane permeability and thereby function as an uncoupler, enhancing electron transport in vitro. Hence, its unspecific interference with photosynthetic membranes in vitro suggests that the inhibitory effect observed on intact cell photosynthesis is caused by uncoupling of phosphorylation.

Crystallization and preliminary X-ray diffraction analysis of l,l-diaminopimelate aminotransferase (DapL) from Chlamydomonas reinhardtii

International Nuclear Information System (INIS)

Hudson, André O.; Girón, Irma; Dobson, Renwick C. J.

2010-01-01

A variant of the diaminopimelate/lysine pathway has recently been defined following the discovery of the enzyme l,l-diaminopimelate aminotransferase (DapL). The cloning of the cDNA, recombinant expression, purification and preliminary diffraction analysis of DapL from the alga C. reinhardtii are presented. In the anabolic synthesis of diaminopimelate and lysine in plants and in some bacteria, the enzyme l,l-diaminopimelate aminotransferase (DapL; EC 2.6.1.83) catalyzes the conversion of tetrahydrodipicolinic acid (THDPA) to l,l-diaminopimelate, bypassing the DapD, DapC and DapE enzymatic steps in the bacterial acyl pathways. Here, the cloning, expression, purification, crystallization and preliminary X-ray diffraction analysis of DapL from the alga Chlamydomonas reinhardtii are presented. Protein crystals were grown in conditions containing 25%(w/v) PEG 3350 and 200 mM lithium sulfate and initially diffracted to ∼1.35 Å resolution. They belonged to space group P2 1 2 1 2 1 , with unit-cell parameters a = 58.9, b = 91.8, c = 162.9 Å. The data were processed to 1.55 Å resolution with an R merge of 0.081, an R p.i.m. of 0.044, an R r.i.m of 0.093 and a V M of 2.28 Å 3 Da −1

Role of metal mixtures (Ca, Cu and Pb) on Cd bioaccumulation and phytochelatin production by Chlamydomonas reinhardtii

International Nuclear Information System (INIS)

Abboud, Pauline; Wilkinson, Kevin J.

2013-01-01

The goal of the study was to determine whether metal uptake and biological effects could be predicted by free ion concentrations when organisms were exposed to Cd and a second metal. Bioaccumulation and algal phytochelatin (PC) concentrations were determined for Chlamydomonas reinhardtii following a 6-h exposure. Bioaccumulation results, after six hours of exposure, showed that Cd uptake decreased in the presence of relatively high concentrations of Ca, Cu and Pb, however, Cd bioaccumulation increased in the presence of ca. equimolar concentrations of Cu. A good correlation was observed between the production of PCs and the amount of metals bioaccumulated for the binary mixtures of Cd–Pb and Cd–Cu, but not the Cd–Ca mixture. Overall, the results suggested that, in the case of mixtures, bioaccumulated metal rather than free ion concentrations would be a better predictor of biological effect. -- Highlights: •Cd bioaccumulation and phytochelatin production were evaluated for metal mixtures. •Bioaccumulated metal rather than free ion was a better predictor of biological effect. •Calcium additions decreased Cd bioaccumulation but increased phytochelatin production. •Copper additions increased Cd bioaccumulation and phytochelatin production. •Lead additions had little effect on either Cd bioaccumulation or phytochelatin production. -- In metal mixtures containing Cd and Ca, Pb or Cu, bioaccumulated metal rather than free ion was a better predictor of biological effect

The nucleobase cation symporter 1 of Chlamydomonas reinhardtii and that of the evolutionarily distant Arabidopsis thaliana display parallel function and establish a plant-specific solute transport profile.

Science.gov (United States)

Schein, Jessica R; Hunt, Kevin A; Minton, Janet A; Schultes, Neil P; Mourad, George S

2013-09-01

The single cell alga Chlamydomonas reinhardtii is capable of importing purines as nitrogen sources. An analysis of the annotated C. reinhardtii genome reveals at least three distinct gene families encoding for known nucleobase transporters. In this study the solute transport and binding properties for the lone C. reinhardtii nucleobase cation symporter 1 (CrNCS1) are determined through heterologous expression in Saccharomyces cerevisiae. CrNCS1 acts as a transporter of adenine, guanine, uracil and allantoin, sharing similar - but not identical - solute recognition specificity with the evolutionary distant NCS1 from Arabidopsis thaliana. The results suggest that the solute specificity for plant NCS1 occurred early in plant evolution and are distinct from solute transport specificities of single cell fungal NCS1 proteins. Copyright © 2013 Elsevier Masson SAS. All rights reserved.

Growth of Chlamydomonas reinhardtii in acetate-free medium when co-cultured with alginate-encapsulated, acetate-producing strains of Synechococcus sp. PCC 7002.

Science.gov (United States)

Therien, Jesse B; Zadvornyy, Oleg A; Posewitz, Matthew C; Bryant, Donald A; Peters, John W

2014-01-01

The model alga Chlamydomonas reinhardtii requires acetate as a co-substrate for optimal production of lipids, and the addition of acetate to culture media has practical and economic implications for algal biofuel production. Here we demonstrate the growth of C. reinhardtii on acetate provided by mutant strains of the cyanobacterium Synechococcus sp. PCC 7002. Optimal growth conditions for co-cultivation of C. reinhardtii with wild-type and mutant strains of Synechococcus sp. 7002 were established. In co-culture, acetate produced by a glycogen synthase knockout mutant of Synechococcus sp. PCC 7002 was able to support the growth of a lipid-accumulating mutant strain of C. reinhardtii defective in starch production. Encapsulation of Synechococcus sp. PCC 7002 using an alginate matrix was successfully employed in co-cultures to limit growth and maintain the stability. The ability of immobilized strains of the cyanobacterium Synechococcus sp. PCC 7002 to produce acetate at a level adequate to support the growth of lipid-accumulating strains of C. reinhartdii offers a potentially practical, photosynthetic alternative to providing exogenous acetate into growth media.

Study of metabolic pathways for hydrogen production in chlamydomonas reinhardtii and transposition on a torus photo bioreactor; Etude des voies metaboliques de production d'hydrogene chez la microalgue Chlamydomonas reinhardtii et transposition en photobioreacteur

Energy Technology Data Exchange (ETDEWEB)

Fouchard, S

2006-04-15

Considering the recent increase in energy consumption. aide associated environmental risks, new trails are followed today to develop the use of clean and renewable alternative energies. In this context hydrogen seems to be a serious solution and this study, based on micro-algae photosynthetic capacities exploitation, will allow to devise a process for hydrogen production from only water and solar energy without greenhouse gas release. The sulphur deprivation protocol on TAP medium, known to lead to hydrogen production in Chlamydomonas reinhardtii species was particularly studied. At the metabolic level, two important phenomena are induced under these conditions: an over-accumulation of the intracellular starch reserves and a simultaneous alteration of the PsII activity which leads to anoxia and Fe-hydrogenase induction, an enzyme with a strong specific activity responsible for the hydrogen production. The contribution of the two electron transfer pathways implied in the hydrogen production process (PsII-dependent and PSII-independent) as well as the importance of the previously accumulated starch were highlighted here. We also investigated the potential for designing autotrophic protocols for hydrogen photoproduction. Various protocols, considered to be relevant, were then transposed on a torus photo-bioreactor, specifically developed in this study and which allows the control of culture parameters as well as the precise measurement of gas release kinetics, in order to obtain first estimates of productivity of the system. Integration of the physical; aspects of the pilot and biological aspects of the process in a model, finally opens new prospects for subject development, in particular for a reasoned optimization of hydrogen production via this double physiology/process approach. (author)

Molecular toxicity of cerium oxide nanoparticles to the freshwater alga Chlamydomonas reinhardtii is associated with supra-environmental exposure concentrations

Science.gov (United States)

Taylor, Nadine S.; Merrifield, Ruth; Williams, Tim D.; Chipman, J. Kevin; Lead, Jamie R.; Viant, Mark R.

2016-01-01

Abstract Ceria nanoparticles (NPs) are widely used as fuel catalysts and consequently are likely to enter the environment. Their potential impacts on. biota at environmentally relevant concentrations, including uptake and toxicity, remain to be elucidated and quantitative data on which to assess risk are sparse. Therefore, a definitive assessment of the molecular and phenotypic effects of ceria NPs was undertaken, using well-characterised mono-dispersed NPs as their toxicity is likely to be higher, enabling a conservative hazard assessment. Unbiased transcriptomics and metabolomics approaches were used to investigate the potential toxicity of tightly constrained 4–5 nm ceria NPs to the unicellular green alga, Chlamydomonas reinhardtii, a sentinel freshwater species. A wide range of exposure concentrations were investigated from predicted environmental levels, to support hazard assessment, to supra-environmental levels to provide insight into molecular toxicity pathways. Ceria NPs were internalised into intracellular vesicles within C. reinhardtii, yet caused no significant effect on algal growth at any exposure concentration. Molecular perturbations were only detected at supra-environmental ceria NP-concentrations, primarily down-regulation of photosynthesis and carbon fixation with associated effects on energy metabolism. For acute exposures to small mono-dispersed particles, it can be concluded there should be little concern regarding their dispersal into the environment for this trophic level. PMID:25740379

Dissecting the sequential assembly and localization of intraflagellar transport particle complex B in Chlamydomonas.

Directory of Open Access Journals (Sweden)

Elizabeth A Richey

Full Text Available Intraflagellar transport (IFT, the key mechanism for ciliogenesis, involves large protein particles moving bi-directionally along the entire ciliary length. IFT particles contain two large protein complexes, A and B, which are constructed with proteins in a core and several peripheral proteins. Prior studies have shown that in Chlamydomonas reinhardtii, IFT46, IFT52, and IFT88 directly interact with each other and are in a subcomplex of the IFT B core. However, ift46, bld1, and ift88 mutants differ in phenotype as ift46 mutants are able to form short flagella, while the other two lack flagella completely. In this study, we investigated the functional differences of these individual IFT proteins contributing to complex B assembly, stability, and basal body localization. We found that complex B is completely disrupted in bld1 mutant, indicating an essential role of IFT52 for complex B core assembly. Ift46 mutant cells are capable of assembling a relatively intact complex B, but such complex is highly unstable and prone to degradation. In contrast, in ift88 mutant cells the complex B core still assembles and remains stable, but the peripheral proteins no longer attach to the B core. Moreover, in ift88 mutant cells, while complex A and the anterograde IFT motor FLA10 are localized normally to the transition fibers, complex B proteins instead are accumulated at the proximal ends of the basal bodies. In addition, in bld2 mutant, the IFT complex B proteins still localize to the proximal ends of defective centrioles which completely lack transition fibers. Taken together, these results revealed a step-wise assembly process for complex B, and showed that the complex first localizes to the proximal end of the centrioles and then translocates onto the transition fibers via an IFT88-dependent mechanism.

Mechanistic modeling of sulfur-deprived photosynthesis and hydrogen production in suspensions of Chlamydomonas reinhardtii.

Science.gov (United States)

Williams, C R; Bees, M A

2014-02-01

The ability of unicellular green algal species such as Chlamydomonas reinhardtii to produce hydrogen gas via iron-hydrogenase is well known. However, the oxygen-sensitive hydrogenase is closely linked to the photosynthetic chain in such a way that hydrogen and oxygen production need to be separated temporally for sustained photo-production. Under illumination, sulfur-deprivation has been shown to accommodate the production of hydrogen gas by partially-deactivating O2 evolution activity, leading to anaerobiosis in a sealed culture. As these facets are coupled, and the system complex, mathematical approaches potentially are of significant value since they may reveal improved or even optimal schemes for maximizing hydrogen production. Here, a mechanistic model of the system is constructed from consideration of the essential pathways and processes. The role of sulfur in photosynthesis (via PSII) and the storage and catabolism of endogenous substrate, and thus growth and decay of culture density, are explicitly modeled in order to describe and explore the complex interactions that lead to H2 production during sulfur-deprivation. As far as possible, functional forms and parameter values are determined or estimated from experimental data. The model is compared with published experimental studies and, encouragingly, qualitative agreement for trends in hydrogen yield and initiation time are found. It is then employed to probe optimal external sulfur and illumination conditions for hydrogen production, which are found to differ depending on whether a maximum yield of gas or initial production rate is required. The model constitutes a powerful theoretical tool for investigating novel sulfur cycling regimes that may ultimately be used to improve the commercial viability of hydrogen gas production from microorganisms. © 2013 The Authors. Biotechnology and Bioengineering Published by Wiley Periodicals, Inc.

Inorganic polyphosphate occurs in the cell wall of Chlamydomonas reinhardtii and accumulates during cytokinesis

Directory of Open Access Journals (Sweden)

Freimoser Florian M

2007-09-01

Full Text Available Abstract Background Inorganic polyphosphate (poly P, linear chains of phosphate residues linked by energy rich phosphoanhydride bonds, is found in every cell and organelle and is abundant in algae. Depending on its localization and concentration, poly P is involved in various biological functions. It serves, for example, as a phosphate store and buffer against alkali, is involved in energy metabolism and regulates the activity of enzymes. Bacteria defective in poly P synthesis are impaired in biofilm development, motility and pathogenicity. PolyP has also been found in fungal cell walls and bacterial envelopes, but has so far not been measured directly or stained specifically in the cell wall of any plant or alga. Results Here, we demonstrate the presence of poly P in the cell wall of Chlamydomonas reinhardtii by staining with specific poly P binding proteins. The specificity of the poly P signal was verified by various competition experiments, by staining with different poly P binding proteins and by correlation with biochemical quantification. Microscopical investigation at different time-points during growth revealed fluctuations of the poly P signal synchronous with the cell cycle: The poly P staining peaked during late cytokinesis and was independent of the high intracellular poly P content, which fluctuated only slightly during the cell cycle. Conclusion The presented staining method provides a specific and sensitive tool for the study of poly P in the extracellular matrices of algae and could be used to describe the dynamic behaviour of cell wall poly P during the cell cycle. We assume that cell wall poly P and intracellular poly P are regulated by distinct mechanisms and it is suggested that cell wall bound poly P might have important protective functions against toxic compounds or pathogens during cytokinesis, when cells are more vulnerable.

The involvement of carbohydrate reserves in hydrogen photoproduction by the green alga Chlamydomonas reinhardtii

International Nuclear Information System (INIS)

Chochois, V.

2009-09-01

The unicellular green alga Chlamydomonas reinhardtii is able to produce hydrogen, using water as an electron donor, and sunlight as an energy source. Although this property offers interesting biotechnological perspectives, a major limitation is related to the sensitivity of hydrogenase to oxygen which is produced by photosynthesis. It had been previously shown that in conditions of sulfur deprivation, C. reinhardtii is able to produce hydrogen during several days (Melis et an. 2000). During this process, two pathways, one direct depending on photosystem II (PSII) activity and the other involving only the PSI, are involved, starch reserves being supposed to play a role in both of these pathways. The purpose of this phD thesis was to elucidate the mechanisms linking starch catabolism to the hydrogen photoproduction process. Firstly, the analysis of mutants affected in starch biosynthesis (sta6 and sta7) showed that if starch reserves are essential to the functioning of the indirect pathway, they are not involved in the direct one. Secondly, in order to identify metabolic steps and regulatory processes involved in starch breakdown, we developed a genetic approach based on the search of mutants affected in starch reserves mobilization. Eight mutant (std1 to std8) diversely affected in their ability to degrade starch after an accumulation phase have been isolated from an insertional mutant library of 15,000 clones. One of these mutants, std1, is affected in a kinase related to the DYRK family (dual-specificity tyrosine regulated serine threonine kinase). Although the targets of this putative kinase remain to be identified, the analysis of the granule bound proteome displayed profound alterations in the expression profile of starch phosphorylases, potentially involved in starch breakdown. STD1 represents the first starch catabolism regulator identified to date in plants. (author)

Comparison of secretory signal peptides for heterologous protein expression in microalgae: Expanding the secretion portfolio for Chlamydomonas reinhardtii.

Directory of Open Access Journals (Sweden)

João Vitor Dutra Molino

Full Text Available Efficient protein secretion is a desirable trait for any recombinant protein expression system, together with simple, low-cost, and defined media, such as the typical media used for photosynthetic cultures of microalgae. However, low titers of secreted heterologous proteins are usually obtained, even with the most extensively studied microalga Chlamydomonas reinhardtii, preventing their industrial application. In this study, we aimed to expand and evaluate secretory signal peptides (SP for heterologous protein secretion in C. reinhardtii by comparing previously described SP with untested sequences. We compared the SPs from arylsulfatase 1 and carbonic anhydrase 1, with those of untried SPs from binding protein 1, an ice-binding protein, and six sequences identified in silico. We identified over 2000 unique SPs using the SignalP 4.0 software. mCherry fluorescence was used to compare the protein secretion of up to 96 colonies for each construct, non-secretion construct, and parental wild-type cc1690 cells. Supernatant fluorescence varied according to the SP used, with a 10-fold difference observed between the highest and lowest secretors. Moreover, two SPs identified in silico secreted the highest amount of mCherry. Our results demonstrate that the SP should be carefully selected and that efficient sequences can be coded in the C. reinhardtii genome. The SPs described here expand the portfolio available for research on heterologous protein secretion and for biomanufacturing applications.

NRT2.4 and NRT2.5 Are Two Half-Size Transporters from the Chlamydomonas NRT2 Family

Directory of Open Access Journals (Sweden)

Jose Javier Higuera

2016-03-01

Full Text Available The NRT2 transporters mediate High Affinity Nitrate/NitriteTransport (HAN/NiT, which are essential for nitrogen acquisition from these inorganic forms. The NRT2 proteins are encoded by a multigene family in plants, and contain 12 transmembrane-spanning domains. Chlamydomonas reinhardtii has six NRT2, two of which (NRT2.5 and NRT2.4 are located in Chromosome III, in tandem head to tail. cDNAs for these genes were isolated and their sequence revealed that they correspond to half-size NRT2 transporters each containing six transmembrane domains. NRT2.5 has long N- and C- termini sequences without known homology. NRT2.4 also contains long termini sequences but smaller than NRT2.5. Expression of both studied genes occurred at a very low level, slightly in darkness, and was not modified by the N or C source. Silencing of NRT2.4 by specific artificial miRNA resulted in the inhibition of nitrite transport in the absence of other HANNiT (NRT2.1/NAR2 in the cell genetic background. Nitrite transport activity in the Hansenula polymorpha Δynt::URA3 Leu2 mutant was restored by expressing CrNRT2.4. These results indicate that half-size NRT2 transporters are present in photosynthetic organisms and that NRT2.4 is a HANiT.

Determination of the speciation and bioavailability of samarium to Chlamydomonas reinhardtii in the presence of natural organic matter.

Science.gov (United States)

Rowell, Justine-Anne; Fillion, Marc-Alexandre; Smith, Scott; Wilkinson, Kevin J

2018-06-01

As technological interest and environmental emissions of the rare earth elements increase, it is becoming more important to assess their potential environmental impact. Samarium (Sm) is a lanthanide of intermediate molar mass that is used in numerous high-technology applications including wind turbines, solar panels, and electric vehicles. The present study relates the speciation of Sm determined in the presence of natural organic matter (NOM) to its bioavailability to the unicellular green alga Chlamydomonas reinhardtii. The free ion concentration was determined using a cation exchange resin (ion exchange technique) in dynamic mode and compared with thermodynamic modeling. Short-term biouptake experiments were performed in the presence of 4 types of NOM: Suwannee River fulvic acids, Pahokee Peat fulvic acids, Suwannee River humic acids, and a Luther Marsh dissolved organic matter isolate (90-95% humic acids). It was clearly shown that even a small amount of NOM (0.5 mg C L -1 ) resulted in a significant decrease (10 times) in the Sm internalization fluxes. Furthermore, complexation with humic acids (and the corresponding reduction in Sm bioavailability) was stronger than that with fulvic acids. The results showed that the experimentally measured (free) Sm was a better predictor of Sm internalization than either the total concentrations or the free ion concentrations obtained using thermodynamic modeling. Environ Toxicol Chem 2018;37:1623-1631. © 2018 SETAC. © 2018 SETAC.

Biogenesis and fate of the cell-cell adhesion molecule, agglutinin, during gametogenesis and fertilization of Chlamydomonas reinhardtii

International Nuclear Information System (INIS)

Hunnicutt, G.R.

1989-01-01

Fertilization in Chlamydomonas begins with the species-specific recognition and adhesion between gametes of opposite mating types via agglutinin molecules on the flagellar surface. This adhesion generates a cAMP-mediated sexual signal that initiates the subsequent events of call wall release, mating structure activation, and cell fusion. Although flagella of paired gametes remain attached to each other until the zygote forms, the process is dynamic. Engaged agglutinins rapidly become inactivated and turnover, requiring the constant supply of new agglutinins to replace the lost molecules. A population of cell body associated agglutinins has been postulated to the pool of agglutinins recruited during this turnover. Cell body agglutinins, therefore were identified, purified, localized within the cells and compared to flagellar agglutinins. The relationship between these two agglutinin populations was also examined. Cell body agglutinins were biochemically indistinguishable from the flagellar form with respect to their M r , sedimentation coefficient, and hydrophobicity elution properties. Functionally, however, these molecules were inactive in situ. The calculated surface density of agglutinins in the cell body and flagellar domains was similar and thus could not explain their functional difference, but two domains contiguous and yet distinctive suggested they may be separated by a functional barrier. To test this, a method was developed, using a monoclonal antibody and cycloheximide, that removed the flagellar agglutinins so movement between the domains could be monitored. Mobilization of agglutinins onto the flagella did not occur unless sexual signaling was induced with cAMP and papaverine

Study of metabolic pathways for hydrogen production in chlamydomonas reinhardtii and transposition on a torus photo bioreactor; Etude des voies metaboliques de production d'hydrogene chez la microalgue Chlamydomonas reinhardtii et transposition en photobioreacteur

Energy Technology Data Exchange (ETDEWEB)

Fouchard, S

2006-04-15

Considering the recent increase in energy consumption. aide associated environmental risks, new trails are followed today to develop the use of clean and renewable alternative energies. In this context hydrogen seems to be a serious solution and this study, based on micro-algae photosynthetic capacities exploitation, will allow to devise a process for hydrogen production from only water and solar energy without greenhouse gas release. The sulphur deprivation protocol on TAP medium, known to lead to hydrogen production in Chlamydomonas reinhardtii species was particularly studied. At the metabolic level, two important phenomena are induced under these conditions: an over-accumulation of the intracellular starch reserves and a simultaneous alteration of the PsII activity which leads to anoxia and Fe-hydrogenase induction, an enzyme with a strong specific activity responsible for the hydrogen production. The contribution of the two electron transfer pathways implied in the hydrogen production process (PsII-dependent and PSII-independent) as well as the importance of the previously accumulated starch were highlighted here. We also investigated the potential for designing autotrophic protocols for hydrogen photoproduction. Various protocols, considered to be relevant, were then transposed on a torus photo-bioreactor, specifically developed in this study and which allows the control of culture parameters as well as the precise measurement of gas release kinetics, in order to obtain first estimates of productivity of the system. Integration of the physical; aspects of the pilot and biological aspects of the process in a model, finally opens new prospects for subject development, in particular for a reasoned optimization of hydrogen production via this double physiology/process approach. (author)

Combined Increases in Mitochondrial Cooperation and Oxygen Photoreduction Compensate for Deficiency in Cyclic Electron Flow in Chlamydomonas reinhardtii[W][OPEN

Science.gov (United States)

Dang, Kieu-Van; Plet, Julie; Tolleter, Dimitri; Jokel, Martina; Cuiné, Stéphan; Carrier, Patrick; Auroy, Pascaline; Richaud, Pierre; Johnson, Xenie; Alric, Jean; Allahverdiyeva, Yagut; Peltier, Gilles

2014-01-01

During oxygenic photosynthesis, metabolic reactions of CO2 fixation require more ATP than is supplied by the linear electron flow operating from photosystem II to photosystem I (PSI). Different mechanisms, such as cyclic electron flow (CEF) around PSI, have been proposed to participate in reequilibrating the ATP/NADPH balance. To determine the contribution of CEF to microalgal biomass productivity, here, we studied photosynthesis and growth performances of a knockout Chlamydomonas reinhardtii mutant (pgrl1) deficient in PROTON GRADIENT REGULATION LIKE1 (PGRL1)–mediated CEF. Steady state biomass productivity of the pgrl1 mutant, measured in photobioreactors operated as turbidostats, was similar to its wild-type progenitor under a wide range of illumination and CO2 concentrations. Several changes were observed in pgrl1, including higher sensitivity of photosynthesis to mitochondrial inhibitors, increased light-dependent O2 uptake, and increased amounts of flavodiiron (FLV) proteins. We conclude that a combination of mitochondrial cooperation and oxygen photoreduction downstream of PSI (Mehler reactions) supplies extra ATP for photosynthesis in the pgrl1 mutant, resulting in normal biomass productivity under steady state conditions. The lower biomass productivity observed in the pgrl1 mutant in fluctuating light is attributed to an inability of compensation mechanisms to respond to a rapid increase in ATP demand. PMID:24989042

Effect of mutagen combined action on Chlamydomonas reinhardtii cells. II. Dependence of lethal effect on mutagen dose and on conditions of cultivation following mutagen action. [In Slovak

Energy Technology Data Exchange (ETDEWEB)

Podstavkova, S; Vlcek, D; Dubovsky, J [Komenskeho Univ., Bratislava (Czechoslovakia). Prirodovedecka Fakulta

1978-01-01

The effect of UV radiation and UV radiation combined with alkylnitrosourea derivatives (N-methyl-N-nitrosourea and N-ethyl-N-nitrosourea) was observed on survival of cells of the algae Chlamydomonas reinhardtii. In particular, single parts were evaluated of the overall lethal effect - dying of cells before division and dying of cells after division. It was found that the combined action of low doses of UV radiation and alkylnitrosoureas result in a pronounced protective effect which manifests itself by a higher frequency of surviving cells than was that effected by the action of alkylnitrosoureas alone. As a result of combined action with higher doses of UV radiation this effect is lost, and the resultant values will come close to the theoretically anticipated values. This gradual transition from a protective to an additive effect mainly manifests itself by changes in the proportion of cells dying before division.

The microalga Chlamydomonas reinhardtii CW-15 as a solar cell for hydrogen peroxide photoproduction. Comparison between free and immobilized cells and thylakoids for energy conversion efficiency

Energy Technology Data Exchange (ETDEWEB)

Scholz, W.; Galvan, F.; Rosa, F.F. de la [Instituto de Bioquimica Vegetal y Fotosintesis, Universidad de Sevilla y CSIC, Sevilla (Spain)

1995-11-28

Immobilized cells and thylakoid vesicles of the microalga Chlamydomonas reinhardtii CW-15 have been developed as a solar cell because of their capabilities of producing hydrogen peroxide. This compound is an efficient and clean fuel used for rocket propulsion, motors and for heating. Hydrogen peroxide is produced by the photosystem in a catalyst cycle in which a redox mediator (methyl viologen) is reduced by electrons obtained from water by the photosynthetic apparatus of the microalga and it is re-oxidized by the oxygen dissolved in the solution. The photoproduction has been investigated using a discontinuous system with whole cells, or thylakoid vesicles, free or immobilized on alginate. The stimulation by azide as an inhibitor of catalase has also been analyzed. Under determined optimum conditions, the photoproduction by Ca-alginate entrapped cells, with a rate of 33 {mu}mol H{sub 2}O{sub 2}/mg Chl.h, was maintained for several hours with an energy conversion efficiency of 0.25%

Lysis of Chlamydomonas reinhardtii by high-intensity focused ultrasound as a function of exposure time.

Science.gov (United States)

Bigelow, Timothy A; Xu, Jin; Stessman, Dan J; Yao, Linxing; Spalding, Martin H; Wang, Tong

2014-05-01

Efficient lysis of microalgae for lipid extraction is an important concern when processing biofuels. Historically, ultrasound frequencies in the range of 10-40 kHz have been utilized for this task. However, greater efficiencies might be achievable if higher frequencies could be used. In our study, we evaluated the potential of using 1.1 MHz ultrasound to lyse microalgae for biofuel production while using Chlamydomonas reinhardtii as a model organism. The ultrasound was generated using a spherically focused transducer with a focal length of 6.34 cm and an active diameter of 6.36 cm driven by 20 cycle sine-wave tone bursts at a pulse repetition frequency of 2 kHz (3.6% duty cycle). The time-average acoustic power output was 26.2 W while the spatial-peak-pulse-average intensity (ISPPA) for each tone burst was 41 kW/cm(2). The peak compressional and rarefactional pressures at the focus were 102 and 17 MPa, respectively. The exposure time was varied for the different cases in the experiments from 5s to 9 min and cell lysis was assessed by quantifying the percentage of protein and chlorophyll release into the supernate as well as the lipid extractability. Free radical generation and lipid oxidation for the different ultrasound exposures were also determined. We found that there was a statistically significant increase in lipid extractability for all of the exposures compared to the control. The longer exposures also completely fragmented the cells releasing almost all of the protein and chlorophyll into the supernate. The cavitation activity did not significantly increase lipid oxidation while there was a minor trend of increased free radical production with increased ultrasound exposure. Copyright © 2013 Elsevier B.V. All rights reserved.

Sensitivity of the green algae Chlamydomonas reinhardtii to gamma radiation: Photosynthetic performance and ROS formation.

Science.gov (United States)

Gomes, Tânia; Xie, Li; Brede, Dag; Lind, Ole-Christian; Solhaug, Knut Asbjørn; Salbu, Brit; Tollefsen, Knut Erik

2017-02-01

The aquatic environment is continuously exposed to ionizing radiation from both natural and anthropogenic sources, making the characterization of ecological and health risks associated with radiation of large importance. Microalgae represent the main source of biomass production in the aquatic ecosystem, thus becoming a highly relevant biological model to assess the impacts of gamma radiation. However, little information is available on the effects of gamma radiation on microalgal species, making environmental radioprotection of this group of species challenging. In this context, the present study aimed to improve the understanding of the effects and toxic mechanisms of gamma radiation in the unicellular green algae Chlamydomonas reinhardtii focusing on the activity of the photosynthetic apparatus and ROS formation. Algal cells were exposed to gamma radiation (0.49-1677mGy/h) for 6h and chlorophyll fluorescence parameters obtained by PAM fluorometry, while two fluorescent probes carboxy-H 2 DFFDA and DHR 123 were used for the quantification of ROS. The alterations seen in functional parameters of C. reinhardtii PSII after 6h of exposure to gamma radiation showed modifications of PSII energy transfer associated with electron transport and energy dissipation pathways, especially at the higher dose rates used. Results also showed that gamma radiation induced ROS in a dose-dependent manner under both light and dark conditions. The observed decrease in photosynthetic efficiency seems to be connected to the formation of ROS and can potentially lead to oxidative stress and cellular damage in chloroplasts. To our knowledge, this is the first report on changes in several chlorophyll fluorescence parameters associated with photosynthetic performance and ROS formation in microalgae after exposure to gamma radiation. Copyright © 2016 Elsevier B.V. All rights reserved.

Functional analysis of three type-2 DGAT homologue genes for triacylglycerol production in the green microalga Chlamydomonas reinhardtii.

Science.gov (United States)

La Russa, M; Bogen, C; Uhmeyer, A; Doebbe, A; Filippone, E; Kruse, O; Mussgnug, J H

2012-11-30

Photosynthetic organisms like plants and algae can use sunlight to produce lipids as important metabolic compounds. Plant-derived triacylglycerols (TAGs) are valuable for human and animal nutrition because of their high energy content and are becoming increasingly important for the production of renewable biofuels. Acyl-CoA:diacylglycerol acyltransferases (DGATs) have been demonstrated to play an important role in the accumulation of TAG compounds in higher plants. DGAT homologue genes have been identified in the genome of the green alga Chlamydomonas reinhardtii, however their function in vivo is still unknown. In this work, the three most promising type-2 DGAT candidate genes potentially involved in TAG lipid accumulation (CrDGAT2a, b and c) were investigated by constructing overexpression strains. For each of the genes, three strains were identified which showed enhanced mRNA levels of between 1.7 and 29.1 times that of the wild type (wt). Total lipid contents, neutral lipids and fatty acid profiles were determined and showed that an enhanced mRNA expression level of the investigated DGAT genes did not boost the intracellular TAG accumulation or resulted in alterations of the fatty acid profiles compared to wild type during standard growth condition or during nitrogen or sulfur stress conditions. We conclude that biotechnological efforts to enhance cellular TAG amount in microalgae need further insights into the complex network of lipid biosynthesis to identify potential bottlenecks of neutral lipid production. Copyright © 2012 Elsevier B.V. All rights reserved.

Absorption and emission spectroscopic characterisation of combined wildtype LOV1-LOV2 domain of phot from Chlamydomonas reinhardtii.

Science.gov (United States)

Song, S-H; Dick, B; Zirak, P; Penzkofer, A; Schiereis, T; Hegemann, P

2005-10-03

An absorption and emission spectroscopic characterisation of the combined wild-type LOV1-LOV2 domain string (abbreviated LOV1/2) of phot from the green alga Chlamydomonas reinhardtii is carried out at pH 8. A LOV1/2-MBP fusion protein (MBP=maltose binding protein) and LOV1/2 with a His-tag at the C-terminus (LOV1/2-His) expressed in an Escherichia coli strain are investigated. Blue-light photo-excitation generates a non-fluorescent intermediate photoproduct (flavin-C(4a)-cysteinyl adduct with absorption peak at 390 nm). The photo-cycle dynamics is studied by dark-state absorption and fluorescence measurement, by following the temporal absorption and emission changes under blue and violet light exposure, and by measuring the temporal absorption and fluorescence recovery after light exposure. The fluorescence quantum yield, phi(F), of the dark adapted samples is phi(F)(LOV1/2-His) approximately 0.15 and phi(F)(LOV1/2-MBP) approximately 0.17. A bi-exponential absorption recovery after light exposure with a fast (in the several 10-s range) and a slow component (in the near 10-min range) are resolved. The quantum yield of photo-adduct formation, phi(Ad), is extracted from excitation intensity dependent absorption measurements. It decreases somewhat with rising excitation intensity. The behaviour of the combined wildtype LOV1-LOV2 double domains is compared with the behaviour of the separate LOV1 and LOV2 domains.

Sensitivity evaluation of the green alga Chlamydomonas reinhardtii to uranium by pulse amplitude modulated (PAM) fluorometry.

Science.gov (United States)

Herlory, Olivier; Bonzom, Jean-Marc; Gilbin, Rodolphe

2013-09-15

Although ecotoxicological studies tend to address the toxicity thresholds of uranium in freshwaters, there is a lack of information on the effects of the metal on physiological processes, particularly in aquatic plants. Knowing that uranium alters photosynthesis via impairment of the water photo-oxidation process, we determined whether pulse amplitude modulated (PAM) fluorometry was a relevant tool for assessing the impact of uranium on the green alga Chlamydomonas reinhardtii and investigated how and to what extent uranium hampered photosynthetic performance. Photosynthetic activity and quenching were assessed from fluorescence induction curves generated by PAM fluorometry, after 1 and 5h of uranium exposure in controlled conditions. The oxygen-evolving complex (OEC) of PSII was identified as the primary action site of uranium, through alteration of the water photo-oxidation process as revealed by F0/Fv. Limiting re-oxidation of the plastoquinone pool, uranium impaired the electron flux between the photosystems until almost complete inhibition of the PSII quantum efficiency ( [Formula: see text] , EC50=303 ± 64 μg UL(-1) after 5h of exposure) was observed. Non-photochemical quenching (qN) was identified as the most sensitive fluorescence parameter (EC50=142 ± 98 μg UL(-1) after 5h of exposure), indicating that light energy not used in photochemistry was dissipated in non-radiative processes. It was shown that parameters which stemmed from fluorescence induction kinetics are valuable indicators for evaluating the impact of uranium on PSII in green algae. PAM fluorometry provided a rapid and reasonably sensitive method for assessing stress response to uranium in microalgae. Copyright © 2013 Elsevier B.V. All rights reserved.

Multiple-endpoint assay provides a detailed mechanistic view of responses to herbicide exposure in Chlamydomonas reinhardtii

International Nuclear Information System (INIS)

Nestler, Holger; Groh, Ksenia J.; Schönenberger, René; Behra, Renata; Schirmer, Kristin; Eggen, Rik I.L.; Suter, Marc J.-F.

2012-01-01

The release of herbicides into the aquatic environment raises concerns about potential detrimental effects on ecologically important non-target species, such as unicellular algae, necessitating ecotoxicological risk assessment. Algal toxicity tests based on growth, a commonly assessed endpoint, are integrative, and hence do not provide information about underlying toxic mechanisms and effects. This limitation may be overcome by measuring more specific biochemical and physiological endpoints. In the present work, we developed and applied a novel multiple-endpoint assay, and analyzed the effects of the herbicides paraquat, diuron and norflurazon, each representing a specific mechanism of toxic action, on the single celled green alga Chlamydomonas reinhardtii. The endpoints added to assessment of growth were pigment content, maximum and effective photosystem II quantum yield, ATP content, esterase and oxidative activity. All parameters were measured at 2, 6 and 24 h of exposure, except for growth and pigment content, which were determined after 6 and 24 h only. Effective concentrations causing 50% of response (EC50s) and lowest observable effect concentrations (LOECs) were determined for all endpoints and exposure durations where possible. The assay provided a detailed picture of the concentration- and time-dependent development of effects elicited by the analyzed herbicides, thus improving the understanding of the underlying toxic mechanisms. Furthermore, the response patterns were unique to the respective herbicide and reflected the different mechanisms of toxicity. The comparison of the endpoint responses and sensitivities revealed that several physiological and biochemical parameters reacted earlier or stronger to disturbances than growth. Overall, the presented multiple-endpoint assay constitutes a promising basis for investigating stressor and toxicant effects in green algae.

Multiple-endpoint assay provides a detailed mechanistic view of responses to herbicide exposure in Chlamydomonas reinhardtii

Energy Technology Data Exchange (ETDEWEB)

Nestler, Holger [Eawag, Swiss Federal Institute of Aquatic Science and Technology, Ueberlandstrasse 133, 8600 Duebendorf (Switzerland); ETH Zurich, Swiss Federal Institute of Technology, Institute of Biogeochemistry and Pollutant Dynamics, Universitaetstrasse 16, 8092 Zurich (Switzerland); Groh, Ksenia J.; Schoenenberger, Rene; Behra, Renata [Eawag, Swiss Federal Institute of Aquatic Science and Technology, Ueberlandstrasse 133, 8600 Duebendorf (Switzerland); Schirmer, Kristin [Eawag, Swiss Federal Institute of Aquatic Science and Technology, Ueberlandstrasse 133, 8600 Duebendorf (Switzerland); ETH Zurich, Swiss Federal Institute of Technology, Institute of Biogeochemistry and Pollutant Dynamics, Universitaetstrasse 16, 8092 Zurich (Switzerland); EPF Lausanne, School of Architecture, Civil and Environmental Engineering, 1015 Lausanne (Switzerland); Eggen, Rik I.L. [Eawag, Swiss Federal Institute of Aquatic Science and Technology, Ueberlandstrasse 133, 8600 Duebendorf (Switzerland); ETH Zurich, Swiss Federal Institute of Technology, Institute of Biogeochemistry and Pollutant Dynamics, Universitaetstrasse 16, 8092 Zurich (Switzerland); Suter, Marc J.-F., E-mail: suter@eawag.ch [Eawag, Swiss Federal Institute of Aquatic Science and Technology, Ueberlandstrasse 133, 8600 Duebendorf (Switzerland); ETH Zurich, Swiss Federal Institute of Technology, Institute of Biogeochemistry and Pollutant Dynamics, Universitaetstrasse 16, 8092 Zurich (Switzerland)

2012-04-15

The release of herbicides into the aquatic environment raises concerns about potential detrimental effects on ecologically important non-target species, such as unicellular algae, necessitating ecotoxicological risk assessment. Algal toxicity tests based on growth, a commonly assessed endpoint, are integrative, and hence do not provide information about underlying toxic mechanisms and effects. This limitation may be overcome by measuring more specific biochemical and physiological endpoints. In the present work, we developed and applied a novel multiple-endpoint assay, and analyzed the effects of the herbicides paraquat, diuron and norflurazon, each representing a specific mechanism of toxic action, on the single celled green alga Chlamydomonas reinhardtii. The endpoints added to assessment of growth were pigment content, maximum and effective photosystem II quantum yield, ATP content, esterase and oxidative activity. All parameters were measured at 2, 6 and 24 h of exposure, except for growth and pigment content, which were determined after 6 and 24 h only. Effective concentrations causing 50% of response (EC50s) and lowest observable effect concentrations (LOECs) were determined for all endpoints and exposure durations where possible. The assay provided a detailed picture of the concentration- and time-dependent development of effects elicited by the analyzed herbicides, thus improving the understanding of the underlying toxic mechanisms. Furthermore, the response patterns were unique to the respective herbicide and reflected the different mechanisms of toxicity. The comparison of the endpoint responses and sensitivities revealed that several physiological and biochemical parameters reacted earlier or stronger to disturbances than growth. Overall, the presented multiple-endpoint assay constitutes a promising basis for investigating stressor and toxicant effects in green algae.

Evidence that an internal carbonic anhydrase is present in 5% CO2-grown and air-grown Chlamydomonas

International Nuclear Information System (INIS)

Moroney, J.V.; Togasaki, R.K.; Husic, H.D.; Tolbert, N.E.

1987-01-01

Inorganic carbon (C/sub i/) uptake was measured in wild-type cells of Chlamydomonas reinhardtii, and in cia-3, a mutant strain of C. reinhardtii that cannot grow with air levels of CO 2 . Both air-grown cells, that have a CO 2 concentrating system, and 5% CO 2 -grown cells that do not have this system, were used. When the external pH was 5.1 or 7.3, air-grown, wild-type cells accumulated inorganic carbon (C/sub i/) and this accumulation was enhanced when the permeant carbonic anhydrase inhibitor, ethoxyzolamide, was added. When the external pH was 5.1, 5% CO 2 -grown cells also accumulated some C/sub i/, although not as much as air-grown cells and this accumulation was stimulated by the addition of ethoxyzolamide. At the same time, ethoxyzolamide inhibited CO 2 fixation by high CO 2 -grown, wild-type cells at both pH 5.1 and 7.3. These observations imply that 5% CO 2 -grown, wild-type cells, have a physiologically important internal carbonic anhydrase, although the major carbonic anhydrase located in the periplasmic space is only present in air-grown cells. Inorganic carbon uptake by cia-3 cells supported this conclusion. This mutant strain, which is thought to lack an internal carbonic anhydrase, was unaffected by ethoxyzolamide at pH 5.1. Other physiological characteristics of cia-3 resemble those of wild-type cells that have been treated with ethoxyzolamide. It is concluded that an internal carbonic anhydrase is under different regulatory control than the periplasmic carbonic anhydrase

Efeito do uso de efluente doméstico tratado, como meio de cultura, sobre o crescimento e produtividade no cultivo de chlamydomonas sp. Isolada de lixiviado de aterro sanitário

Directory of Open Access Journals (Sweden)

Fábio de Farias Neves

2013-01-01

Full Text Available É crescente a aplicação do cultivo de microalgas no campo da Biotecnologia Ambiental, buscando fixação de dióxido de carbono (CO2 e obtenção de energia da biomassa. Entretanto, para essas aplicações se tornarem economicamente viáveis, é necessáriaa adoção de estratégias para baixar o custo de produção de microalgas. A presente pesquisa avaliou o efeito do uso de efluentedoméstico tratado como meio de cultura alternativo de baixo custo sobre o crescimento e a produtividade do cultivo de Chlamydomonas sp. isolada de uma lagoa de tratamento de lixiviados de aterro sanitário, situada na região sul do Brasil. Três tratamentos foram testados: um controle utilizando o meio de cultura sintético TAP, outro com 50% do meio TAP e 50% do efluente e o terceiro com 100% do efluente. Não houve diferença significativa dos parâmetros de crescimento entre os tratamentos, assim como entre a produtividade alcançada nos cultivos com meio TAP e 100% efluente, atingindo valores de massa seca após 10 dias de cultivo de 1,4 ± 0,14g L-1 e 1,3 ± 0,19 g L-1 respectivamente. Já o cultivo em meio TAP com adição de 50% do efluente apresentou a maior produtividade, atingindo um valor de massa seca médio após 10 dias de cultivo de 1,7 ± 0,07 g L-1. Os resultados demonstram que o efluente doméstico tratado tem potencial para ser utilizado como meio de cultura para o cultivo das cepas de Chlamydomonas sp. sem prejudicar o crescimento e a produtividade Abstract Microalgae have been cultured increasingly in order to fix carbon dioxide and produce biofuels from the biomass. However, it is important to develop low cost strategies for microalgae production in order to turn this into a viable alternative of renewable energy. The present investigation studied the effect of treated wastewater used as an alternative culture medium for growth and productivity of a Chlamydomonas sp. strain isolated from landfills leachate of a treatment pond located in

Global Metabolic Regulation of the Snow Alga Chlamydomonas nivalis in Response to Nitrate or Phosphate Deprivation by a Metabolome Profile Analysis.

Science.gov (United States)

Lu, Na; Chen, Jun-Hui; Wei, Dong; Chen, Feng; Chen, Gu

2016-05-10

In the present work, Chlamydomonas nivalis, a model species of snow algae, was used to illustrate the metabolic regulation mechanism of microalgae under nutrient deprivation stress. The seed culture was inoculated into the medium without nitrate or phosphate to reveal the cell responses by a metabolome profile analysis using gas chromatography time-of-flight mass spectrometry (GC/TOF-MS). One hundred and seventy-one of the identified metabolites clustered into five groups by the orthogonal partial least squares discriminant analysis (OPLS-DA) model. Among them, thirty of the metabolites in the nitrate-deprived group and thirty-nine of the metabolites in the phosphate-deprived group were selected and identified as "responding biomarkers" by this metabolomic approach. A significant change in the abundance of biomarkers indicated that the enhanced biosynthesis of carbohydrates and fatty acids coupled with the decreased biosynthesis of amino acids, N-compounds and organic acids in all the stress groups. The up- or down-regulation of these biomarkers in the metabolic network provides new insights into the global metabolic regulation and internal relationships within amino acid and fatty acid synthesis, glycolysis, the tricarboxylic acid cycle (TCA) and the Calvin cycle in the snow alga under nitrate or phosphate deprivation stress.

The Search for a Lipid Trigger: The Effect of Salt Stress on the Lipid Profile of the Model Microalgal Species Chlamydomonas reinhardtii for Biofuels Production.

Science.gov (United States)

Hounslow, Emily; Kapoore, Rahul Vijay; Vaidyanathan, Seetharaman; Gilmour, D James; Wright, Phillip C

2016-11-01

Algal cells produce neutral lipid when stressed and this can be used to generate biodiesel. Salt stressed cells of the model microalgal species Chlamydomonas reinhardtii were tested for their suitability to produce lipid for biodiesel. The starchless mutant of C. reinhardtii (CC-4325) was subjected to salt stress (0.1, 0.2 and 0.3 M NaCl) and transesterification and GC analysis were used to determine fatty acid methyl ester (FAME) content and profile. Fatty acid profile was found to vary under salt stress conditions, with a clear distinction between 0.1 M NaCl, which the algae could tolerate, and the higher levels of NaCl (0.2 and 0.3 M), which caused cell death. Lipid content was increased under salt conditions, either through long-term exposure to 0.1 M NaCl, or short-term exposure to 0.2 and 0.3 M NaCl. Palmitic acid (C16:0) and linolenic acid (C18:3n3) were found to increase significantly at the higher salinities. Salt increase can act as a lipid trigger for C. reinhardtii.

Light-Harvesting Complex Protein LHCBM9 Is Critical for Photosystem II Activity and Hydrogen Production in Chlamydomonas reinhardtii[C][W

Science.gov (United States)

Grewe, Sabrina; Ballottari, Matteo; Alcocer, Marcelo; D’Andrea, Cosimo; Blifernez-Klassen, Olga; Hankamer, Ben; Mussgnug, Jan H.; Bassi, Roberto; Kruse, Olaf

2014-01-01

Photosynthetic organisms developed multiple strategies for balancing light-harvesting versus intracellular energy utilization to survive ever-changing environmental conditions. The light-harvesting complex (LHC) protein family is of paramount importance for this function and can form light-harvesting pigment protein complexes. In this work, we describe detailed analyses of the photosystem II (PSII) LHC protein LHCBM9 of the microalga Chlamydomonas reinhardtii in terms of expression kinetics, localization, and function. In contrast to most LHC members described before, LHCBM9 expression was determined to be very low during standard cell cultivation but strongly increased as a response to specific stress conditions, e.g., when nutrient availability was limited. LHCBM9 was localized as part of PSII supercomplexes but was not found in association with photosystem I complexes. Knockdown cell lines with 50 to 70% reduced amounts of LHCBM9 showed reduced photosynthetic activity upon illumination and severe perturbation of hydrogen production activity. Functional analysis, performed on isolated PSII supercomplexes and recombinant LHCBM9 proteins, demonstrated that presence of LHCBM9 resulted in faster chlorophyll fluorescence decay and reduced production of singlet oxygen, indicating upgraded photoprotection. We conclude that LHCBM9 has a special role within the family of LHCII proteins and serves an important protective function during stress conditions by promoting efficient light energy dissipation and stabilizing PSII supercomplexes. PMID:24706511

Acclimation of Chlamydomonas reinhardtii to ultraviolet radiation and its impact on chemical toxicity

Energy Technology Data Exchange (ETDEWEB)

Korkaric, Muris; Xiao, Mao [Eawag, Swiss Federal Institute of Aquatic Science and Technology, Department of Environmental Toxicology, 8600 Duebendorf (Switzerland); ETH Zürich, Institute of Biogeochemistry and Pollutant Dynamics, 8092 Zürich (Switzerland); Behra, Renata [Eawag, Swiss Federal Institute of Aquatic Science and Technology, Department of Environmental Toxicology, 8600 Duebendorf (Switzerland); Eggen, Rik I.L., E-mail: rik.eggen@eawag.ch [Eawag, Swiss Federal Institute of Aquatic Science and Technology, Department of Environmental Toxicology, 8600 Duebendorf (Switzerland); ETH Zürich, Institute of Biogeochemistry and Pollutant Dynamics, 8092 Zürich (Switzerland)

2015-10-15

Highlights: • Systematic study of UVR acclimation and its impact on chemical toxicity in C. reinhardtii. • UVR acclimation is mediated through fast and reversible physiological defense mechanisms. • Pigment analysis suggests a role of lutein in UVR acclimation. • Co-tolerance to rose bengal suggests a role of singlet oxygen defense in UVR acclimation. • Knowledge on the toxic mechanism of chemicals needed to predict co-tolerance. - Abstract: The toxicity of chemical pollutants can be modulated under stressful environmental conditions, such as increased temperature, salinity or ultraviolet radiation (UVR), due to the interaction of effects during simultaneous stressor exposure. However, organisms may acclimate to such conditions by activation of physiological and biochemical defence mechanisms. In sequential exposures, organisms acclimated to environmental stressors may display an increased sensitivity or co-tolerance towards chemical pollutants. It has been suggested that co-tolerance might be expected for similarly acting stressors due to common defence mechanisms. To test this for combinations of UVR and chemical stressors, we first acclimatized the model green alga Chlamydomonas reinhardtii to UVR and subsequently compared the sensitivity of UVR pre-exposed and control algae towards chemicals. Selected chemicals all act on photosynthesis and thus share a common physiological target, but display distinct toxicity mechanisms. Results showed that UVR pre-exposure for four days partially inhibited algal growth and photosynthesis, but also increased algal tolerance to higher UVR levels, confirming UVR acclimation. HPLC analysis of algal pigments indicated that UVR acclimation might in part be explained by the protective function of lutein while the contribution of UVR absorbing compounds was less clear. Challenge exposure to chemicals in the absence of UVR showed that acclimated algae were co-tolerant to the photosensitizer rose bengal, but not to the

Acclimation of Chlamydomonas reinhardtii to ultraviolet radiation and its impact on chemical toxicity

International Nuclear Information System (INIS)

Korkaric, Muris; Xiao, Mao; Behra, Renata; Eggen, Rik I.L.

2015-01-01

Highlights: • Systematic study of UVR acclimation and its impact on chemical toxicity in C. reinhardtii. • UVR acclimation is mediated through fast and reversible physiological defense mechanisms. • Pigment analysis suggests a role of lutein in UVR acclimation. • Co-tolerance to rose bengal suggests a role of singlet oxygen defense in UVR acclimation. • Knowledge on the toxic mechanism of chemicals needed to predict co-tolerance. - Abstract: The toxicity of chemical pollutants can be modulated under stressful environmental conditions, such as increased temperature, salinity or ultraviolet radiation (UVR), due to the interaction of effects during simultaneous stressor exposure. However, organisms may acclimate to such conditions by activation of physiological and biochemical defence mechanisms. In sequential exposures, organisms acclimated to environmental stressors may display an increased sensitivity or co-tolerance towards chemical pollutants. It has been suggested that co-tolerance might be expected for similarly acting stressors due to common defence mechanisms. To test this for combinations of UVR and chemical stressors, we first acclimatized the model green alga Chlamydomonas reinhardtii to UVR and subsequently compared the sensitivity of UVR pre-exposed and control algae towards chemicals. Selected chemicals all act on photosynthesis and thus share a common physiological target, but display distinct toxicity mechanisms. Results showed that UVR pre-exposure for four days partially inhibited algal growth and photosynthesis, but also increased algal tolerance to higher UVR levels, confirming UVR acclimation. HPLC analysis of algal pigments indicated that UVR acclimation might in part be explained by the protective function of lutein while the contribution of UVR absorbing compounds was less clear. Challenge exposure to chemicals in the absence of UVR showed that acclimated algae were co-tolerant to the photosensitizer rose bengal, but not to the

Rubisco activase is required for optimal photosynthesis in the green alga Chlamydomonas reinhardtii in a low-CO(2) atmosphere.

Science.gov (United States)

Pollock, Steve V; Colombo, Sergio L; Prout, Davey L; Godfrey, Ashley C; Moroney, James V

2003-12-01

This report describes a Chlamydomonas reinhardtii mutant that lacks Rubisco activase (Rca). Using the BleR (bleomycin resistance) gene as a positive selectable marker for nuclear transformation, an insertional mutagenesis screen was performed to select for cells that required a high-CO2 atmosphere for optimal growth. The DNA flanking the BleR insert of one of the high-CO2-requiring strains was cloned using thermal asymmetric interlaced-polymerase chain reaction and inverse polymerase chain reaction and sequenced. The flanking sequence matched the C. reinhardtii Rca cDNA sequence previously deposited in the National Center for Biotechnology Information database. The loss of a functional Rca in the strain was confirmed by the absence of Rca mRNA and protein. The open reading frame for Rca was cloned and expressed in pSL18, a C. reinhardtii expression vector conferring paromomycin resistance. This construct partially complemented the mutant phenotype, supporting the hypothesis that the loss of Rca was the reason the mutant grew poorly in a low-CO2 atmosphere. Sequencing of the C. reinhardtii Rca gene revealed that it contains 10 exons ranging in size from 18 to 470 bp. Low-CO2-grown rca1 cultures had a growth rate and maximum rate of photosynthesis 60% of wild-type cells. Results obtained from experiments on a cia5 rca1 double mutant also suggest that the CO2-concentrating mechanism partially compensates for the absence of an active Rca in the green alga C. reinhardtii.

Chlamydomonas chloroplasts can use short dispersed repeats and multiple pathways to repair a double-strand break in the genome.

Science.gov (United States)

Odom, Obed W; Baek, Kwang-Hyun; Dani, Radhika N; Herrin, David L

2008-03-01

Certain group I introns insert into intronless DNA via an endonuclease that creates a double-strand break (DSB). There are two models for intron homing in phage: synthesis-dependent strand annealing (SDSA) and double-strand break repair (DSBR). The Cr.psbA4 intron homes efficiently from a plasmid into the chloroplast psbA gene in Chlamydomonas, but little is known about the mechanism. Analysis of co-transformants selected using a spectinomycin-resistant 16S gene (16S(spec)) provided evidence for both pathways. We also examined the consequences of the donor DNA having only one-sided or no homology with the psbA gene. When there was no homology with the donor DNA, deletions of up to 5 kb involving direct repeats that flank the psbA gene were obtained. Remarkably, repeats as short as 15 bp were used for this repair, which is consistent with the single-strand annealing (SSA) pathway. When the donor had one-sided homology, the DSB in most co-transformants was repaired using two DNAs, the donor and the 16S(spec) plasmid, which, coincidentally, contained a region that is repeated upstream of psbA. DSB repair using two separate DNAs provides further evidence for the SDSA pathway. These data show that the chloroplast can repair a DSB using short dispersed repeats located proximally, distally, or even on separate molecules relative to the DSB. They also provide a rationale for the extensive repertoire of repeated sequences in this genome.

Multiple stressor effects in Chlamydomonas reinhardtii--toward understanding mechanisms of interaction between effects of ultraviolet radiation and chemical pollutants.

Science.gov (United States)

Korkaric, Muris; Behra, Renata; Fischer, Beat B; Junghans, Marion; Eggen, Rik I L

2015-05-01

The effects of chemical pollutants and environmental stressors, such as ultraviolet radiation (UVR), can interact when organisms are simultaneously exposed, resulting in higher (synergistic) or lower (antagonistic) multiple stressor effects than expected based on the effects of single stressors. Current understanding of interactive effects is limited due to a lack of mechanism-based multiple stressor studies. It has been hypothesized that effect interactions may generally occur if chemical and non-chemical stressors cause similar physiological effects in the organism. To test this hypothesis, we exposed the model green alga Chlamydomonas reinhardtii to combinations of UVR and single chemicals displaying modes of action (MOA) similar or dissimilar to the impact of UVR on photosynthesis. Stressor interactions were analyzed based on the independent action model. Effect interactions were found to depend on the MOA of the chemicals, and also on their concentrations, the exposure time and the measured endpoint. Indeed, only chemicals assumed to cause effects on photosynthesis similar to UVR showed interactions with UVR on photosynthetic yield: synergistic in case of Cd(II) and paraquat and antagonistic in case of diuron. No interaction on photosynthesis was observed for S-metolachlor, which acts dissimilarly to UVR. However, combined effects of S-metolachlor and UVR on algal reproduction were synergistic, highlighting the importance of considering additional MOA of UVR. Possible mechanisms of stressor effect interactions are discussed. Copyright © 2015 Elsevier B.V. All rights reserved.

Expression of type 2 diacylglycerol acyltransferse gene DGTT1 from Chlamydomonas reinhardtii enhances lipid production in Scenedesmus obliquus.

Science.gov (United States)

Chen, Chun-Yen; Kao, Ai-Ling; Tsai, Zheng-Chia; Chow, Te-Jin; Chang, Hsin-Yueh; Zhao, Xin-Qing; Chen, Po-Ting; Su, Hsiang-Yen; Chang, Jo-Shu

2016-03-01

Microalgal strains of Scenedesmus obliquus have the great potential for the production of biofuels, CO2 fixation, and bioremediation. However, metabolic engineering of S. obliquus to improve their useful phenotypes are still not fully developed. In this study, S. obliquus strain CPC2 was genetically engineered to promote the autotrophic growth and lipid productivity. The overexpression plasmid containing the type 2 diacylglycerol acyltransferse (DGAT) gene DGTT1 from Chlamydomonas reinhardtii was constructed and transformed into S. obliquus CPC2, and the positive transformants were obtained. The expression of DGTT1 gene was confirmed by reverse transcription PCR analysis. Enhanced lipid content of the transformant S. obliquus CPC2-G1 by nearly two-fold was observed. The biomass concentration of the recombinant strains was also 29% higher than that of the wild-type strain. Furthermore, the recombinant strain CPC2-G1 was successfully grown in 40 L tubular type photobioreactor and open pond system in an outdoor environment. The lipid content, biomass concentration, and biomass productivity obtained from 40 L tubular PBR were 127.8% 20.0%, and 232.6% higher than those obtained from the wild-type strain. The major aim of this work is to develop a tool to genetically engineer an isolated S. obliquus strain for the desired purpose. This is the first report that genetic engineering of S. obliquus has been successful employed to improve both the microalgal cell growth and the lipid production. Copyright © 2016 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

Toxicity and mode of action of tritium alone and mixed with copper on the green algae Chlamydomonas reinhardtii

International Nuclear Information System (INIS)

Rety, Celine

2010-01-01

Liquid releases by Nuclear Power Plants (NPP) are composed of a mixture of radioactive and non-radioactive substances. When organisms are exposed to mixtures of contaminants the resultant toxicity can be enhanced, or reduced, due to interactions. In order to identify potential interactions between substances released by NPP, two substances representative of such effluents (in term of toxicity and of quantity) were selected for studies: Tritiated water (HTO) and copper (Cu). Effects of this binary mixture were studied on the unicellular green algae Chlamydomonas reinhardtii. HTO, when examined along, was not very toxic to C. reinhardtii. The most sensitive and early effect of HTO was an increase in oxidative stress at concentrations of 40 kBq mL -1 (0.13 μGy h -1 ). Algae exposure to the binary mixture HTO/Cu induced interactive effects on oxidative stress. Reactive Oxygen Species production was higher from exposure to the mixture of contaminants than the addition of the effect from each substance individually. This interaction was explained by an enhanced copper uptake by the algae when in the presence of HTO. The observed supra-additive effect could also be due to direct toxic interactions, especially on the antioxidant system. To conclude, this study showed that the effects of a mixture of radioactive and nonradioactive substances can be greater than what would be predicted based on mere addition of individual effects. Even thought this binary mixture is just a small part of NPP effluents, the study showed that potential interactions should be considered when determining ecological risks to aquatic ecosystems from NPP effluents. (author)

Study of metabolic pathways for hydrogen production in chlamydomonas reinhardtii and transposition on a torus photo bioreactor

International Nuclear Information System (INIS)

Fouchard, S.

2006-04-01

Considering the recent increase in energy consumption. aide associated environmental risks, new trails are followed today to develop the use of clean and renewable alternative energies. In this context hydrogen seems to be a serious solution and this study, based on micro-algae photosynthetic capacities exploitation, will allow to devise a process for hydrogen production from only water and solar energy without greenhouse gas release. The sulphur deprivation protocol on TAP medium, known to lead to hydrogen production in Chlamydomonas reinhardtii species was particularly studied. At the metabolic level, two important phenomena are induced under these conditions: an over-accumulation of the intracellular starch reserves and a simultaneous alteration of the PsII activity which leads to anoxia and Fe-hydrogenase induction, an enzyme with a strong specific activity responsible for the hydrogen production. The contribution of the two electron transfer pathways implied in the hydrogen production process (PsII-dependent and PSII-independent) as well as the importance of the previously accumulated starch were highlighted here. We also investigated the potential for designing autotrophic protocols for hydrogen photoproduction. Various protocols, considered to be relevant, were then transposed on a torus photo-bioreactor, specifically developed in this study and which allows the control of culture parameters as well as the precise measurement of gas release kinetics, in order to obtain first estimates of productivity of the system. Integration of the physical; aspects of the pilot and biological aspects of the process in a model, finally opens new prospects for subject development, in particular for a reasoned optimization of hydrogen production via this double physiology/process approach. (author)

Uphill energy transfer in photosystem I from Chlamydomonas reinhardtii. Time-resolved fluorescence measurements at 77 K.

Science.gov (United States)

Giera, Wojciech; Szewczyk, Sebastian; McConnell, Michael D; Redding, Kevin E; van Grondelle, Rienk; Gibasiewicz, Krzysztof

2018-04-04

Energetic properties of chlorophylls in photosynthetic complexes are strongly modulated by their interaction with the protein matrix and by inter-pigment coupling. This spectral tuning is especially striking in photosystem I (PSI) complexes that contain low-energy chlorophylls emitting above 700 nm. Such low-energy chlorophylls have been observed in cyanobacterial PSI, algal and plant PSI-LHCI complexes, and individual light-harvesting complex I (LHCI) proteins. However, there has been no direct evidence of their presence in algal PSI core complexes lacking LHCI. In order to determine the lowest-energy states of chlorophylls and their dynamics in algal PSI antenna systems, we performed time-resolved fluorescence measurements at 77 K for PSI core and PSI-LHCI complexes isolated from the green alga Chlamydomonas reinhardtii. The pool of low-energy chlorophylls observed in PSI cores is generally smaller and less red-shifted than that observed in PSI-LHCI complexes. Excitation energy equilibration between bulk and low-energy chlorophylls in the PSI-LHCI complexes at 77 K leads to population of excited states that are less red-shifted (by ~ 12 nm) than at room temperature. On the other hand, analysis of the detection wavelength dependence of the effective trapping time of bulk excitations in the PSI core at 77 K provided evidence for an energy threshold at ~ 675 nm, above which trapping slows down. Based on these observations, we postulate that excitation energy transfer from bulk to low-energy chlorophylls and from bulk to reaction center chlorophylls are thermally activated uphill processes that likely occur via higher excitonic states of energy accepting chlorophylls.

Impact of Oxidative Stress on Ascorbate Biosynthesis in Chlamydomonas via Regulation of the VTC2 Gene Encoding a GDP-l-galactose Phosphorylase*

Science.gov (United States)

Urzica, Eugen I.; Adler, Lital N.; Page, M. Dudley; Linster, Carole L.; Arbing, Mark A.; Casero, David; Pellegrini, Matteo; Merchant, Sabeeha S.; Clarke, Steven G.

2012-01-01

The l-galactose (Smirnoff-Wheeler) pathway represents the major route to l-ascorbic acid (vitamin C) biosynthesis in higher plants. Arabidopsis thaliana VTC2 and its paralogue VTC5 function as GDP-l-galactose phosphorylases converting GDP-l-galactose to l-galactose-1-P, thus catalyzing the first committed step in the biosynthesis of l-ascorbate. Here we report that the l-galactose pathway of ascorbate biosynthesis described in higher plants is conserved in green algae. The Chlamydomonas reinhardtii genome encodes all the enzymes required for vitamin C biosynthesis via the l-galactose pathway. We have characterized recombinant C. reinhardtii VTC2 as an active GDP-l-galactose phosphorylase. C. reinhardtii cells exposed to oxidative stress show increased VTC2 mRNA and l-ascorbate levels. Genes encoding enzymatic components of the ascorbate-glutathione system (e.g. ascorbate peroxidase, manganese superoxide dismutase, and dehydroascorbate reductase) are also up-regulated in response to increased oxidative stress. These results indicate that C. reinhardtii VTC2, like its plant homologs, is a highly regulated enzyme in ascorbate biosynthesis in green algae and that, together with the ascorbate recycling system, the l-galactose pathway represents the major route for providing protective levels of ascorbate in oxidatively stressed algal cells. PMID:22393048

Identification of pH-sensing Sites in the Light Harvesting Complex Stress-related 3 Protein Essential for Triggering Non-photochemical Quenching in Chlamydomonas reinhardtii.

Science.gov (United States)

Ballottari, Matteo; Truong, Thuy B; De Re, Eleonora; Erickson, Erika; Stella, Giulio R; Fleming, Graham R; Bassi, Roberto; Niyogi, Krishna K

2016-04-01

Light harvesting complex stress-related 3 (LHCSR3) is the protein essential for photoprotective excess energy dissipation (non-photochemical quenching, NPQ) in the model green algaChlamydomonas reinhardtii Activation of NPQ requires low pH in the thylakoid lumen, which is induced in excess light conditions and sensed by lumen-exposed acidic residues. In this work we have used site-specific mutagenesisin vivoandin vitrofor identification of the residues in LHCSR3 that are responsible for sensing lumen pH. Lumen-exposed protonatable residues, aspartate and glutamate, were mutated to asparagine and glutamine, respectively. By expression in a mutant lacking all LHCSR isoforms, residues Asp(117), Glu(221), and Glu(224)were shown to be essential for LHCSR3-dependent NPQ induction inC. reinhardtii Analysis of recombinant proteins carrying the same mutations refoldedin vitrowith pigments showed that the capacity of responding to low pH by decreasing the fluorescence lifetime, present in the wild-type protein, was lost. Consistent with a role in pH sensing, the mutations led to a substantial reduction in binding the NPQ inhibitor dicyclohexylcarbodiimide. © 2016 by The American Society for Biochemistry and Molecular Biology, Inc.

X-ray dense cellular inclusions in the cells of the green alga Chlamydomonas reinhardtii as seen by soft-x-ray microscopy

International Nuclear Information System (INIS)

Stead, A.D.; Ford, T.W.; Page, A.M.; Brown, J.T.; Meyer-Ilse, W.

1997-01-01

Soft x-rays, having a greater ability to penetrate biological material than electrons, have the potential for producing images of intact, living cells. In addition, by using the so-called open-quotes water windowclose quotes area of the soft x-ray spectrum, a degree of natural contrast is introduced into the image due to differential absorption of the wavelengths by compounds with a high carbon content compared to those with a greater oxygen content. The variation in carbon concentration throughout a cell therefore generates an image which is dependent upon the carbon density within the specimen. Using soft x-ray contact microscopy the authors have previously examined the green alga Chlamydomonas reinhardtii, and the most prominent feature of the cells are the numerous x-ray absorbing spheres, But they were not seen by conventional transmission electron microscopy. Similar structures have also been reported by the Goettingen group using their cryo transmission x-ray microscope at BESSY. Despite the fact that these spheres appear to occupy up to 20% or more of the cell volume when seen by x-ray microscopy, they are not visible by transmission electron microscopy. Given the difficulties and criticisms associated with soft x-ray contact microscopy, the present study was aimed at confirming the existence of these cellular inclusions and learning more of their possible chemical composition

X-ray dense cellular inclusions in the cells of the green alga Chlamydomonas reinhardtii as seen by soft-x-ray microscopy

Energy Technology Data Exchange (ETDEWEB)

Stead, A.D.; Ford, T.W.; Page, A.M. [Univ. of London (United Kingdom); Brown, J.T.; Meyer-Ilse, W. [Ernest Orlando Lawrence Berkeley National Lab., CA (United States)

1997-04-01

Soft x-rays, having a greater ability to penetrate biological material than electrons, have the potential for producing images of intact, living cells. In addition, by using the so-called {open_quotes}water window{close_quotes} area of the soft x-ray spectrum, a degree of natural contrast is introduced into the image due to differential absorption of the wavelengths by compounds with a high carbon content compared to those with a greater oxygen content. The variation in carbon concentration throughout a cell therefore generates an image which is dependent upon the carbon density within the specimen. Using soft x-ray contact microscopy the authors have previously examined the green alga Chlamydomonas reinhardtii, and the most prominent feature of the cells are the numerous x-ray absorbing spheres, But they were not seen by conventional transmission electron microscopy. Similar structures have also been reported by the Goettingen group using their cryo transmission x-ray microscope at BESSY. Despite the fact that these spheres appear to occupy up to 20% or more of the cell volume when seen by x-ray microscopy, they are not visible by transmission electron microscopy. Given the difficulties and criticisms associated with soft x-ray contact microscopy, the present study was aimed at confirming the existence of these cellular inclusions and learning more of their possible chemical composition.

Sensitivity of the green algae Chlamydomonas reinhardtii to gamma radiation: Photosynthetic performance and ROS formation

Energy Technology Data Exchange (ETDEWEB)

Gomes, Tânia, E-mail: tania.gomes@niva.no [Norwegian Institute for Water Research (NIVA), Section of Ecotoxicology and Risk Assessment, Gaustadalléen 21, N-0349, Oslo (Norway); Centre for Environmental Radioactivity, Norwegian University of Life Sciences (NMBU), Post Box 5003, N-1432 Ås (Norway); Xie, Li [Norwegian Institute for Water Research (NIVA), Section of Ecotoxicology and Risk Assessment, Gaustadalléen 21, N-0349, Oslo (Norway); Centre for Environmental Radioactivity, Norwegian University of Life Sciences (NMBU), Post Box 5003, N-1432 Ås (Norway); Brede, Dag; Lind, Ole-Christian [Centre for Environmental Radioactivity, Norwegian University of Life Sciences (NMBU), Post Box 5003, N-1432 Ås (Norway); Department for Environmental Sciences, Faculty of Environmental Science & Technology, Norwegian University of Life Sciences (NMBU), Post Box 5003, N-1432, Ås (Norway); Solhaug, Knut Asbjørn [Centre for Environmental Radioactivity, Norwegian University of Life Sciences (NMBU), Post Box 5003, N-1432 Ås (Norway); Department of Ecology and Natural Resource Management, Norwegian University of Life Sciences (NMBU), Postbox 5003, N-1432, Ås (Norway); Salbu, Brit [Centre for Environmental Radioactivity, Norwegian University of Life Sciences (NMBU), Post Box 5003, N-1432 Ås (Norway); Department for Environmental Sciences, Faculty of Environmental Science & Technology, Norwegian University of Life Sciences (NMBU), Post Box 5003, N-1432, Ås (Norway); and others

2017-02-15

Highlights: • Chlorophyll fluorescence parameters affected at higher dose rates. • Changes in PSII associated with electron transport and energy dissipation pathways. • Dose-dependent ROS production in algae exposed to gamma radiation. • Decrease in photosynthetic efficiency connected to ROS formation. - Abstract: The aquatic environment is continuously exposed to ionizing radiation from both natural and anthropogenic sources, making the characterization of ecological and health risks associated with radiation of large importance. Microalgae represent the main source of biomass production in the aquatic ecosystem, thus becoming a highly relevant biological model to assess the impacts of gamma radiation. However, little information is available on the effects of gamma radiation on microalgal species, making environmental radioprotection of this group of species challenging. In this context, the present study aimed to improve the understanding of the effects and toxic mechanisms of gamma radiation in the unicellular green algae Chlamydomonas reinhardtii focusing on the activity of the photosynthetic apparatus and ROS formation. Algal cells were exposed to gamma radiation (0.49–1677 mGy/h) for 6 h and chlorophyll fluorescence parameters obtained by PAM fluorometry, while two fluorescent probes carboxy-H{sub 2}DFFDA and DHR 123 were used for the quantification of ROS. The alterations seen in functional parameters of C. reinhardtii PSII after 6 h of exposure to gamma radiation showed modifications of PSII energy transfer associated with electron transport and energy dissipation pathways, especially at the higher dose rates used. Results also showed that gamma radiation induced ROS in a dose-dependent manner under both light and dark conditions. The observed decrease in photosynthetic efficiency seems to be connected to the formation of ROS and can potentially lead to oxidative stress and cellular damage in chloroplasts. To our knowledge, this is the first

Uptake of selenium by the unicellular green alga Chlamydomonas reinhardtii - effects induced by chronic exposure

International Nuclear Information System (INIS)

Morlon, H.; Fortin, C.; Pradines, C.; Floriani, M.; Grasset, G.; Adam, C.; Garnier-Laplace, J.

2004-01-01

79 Se is a long-lived radionuclide present in radioactive waste storages. The stable isotope selenium is an essential micro-nutrient that can act against oxidative damage. It is however well known for its bio-magnification potential and chemical toxicity to aquatic life. One of its particularity is to form oxyanions in freshwater ecosystems, which leads to specific behaviours towards biological membranes. Our study deals with the interactions between selenite -Se(IV)- and Chlamydomonas reinhardtii, a unicellular green alga representative of the freshwater phytoplankton community. Cells were exposed to selenite marked with Se 75 in well-known simple inorganic media. Short-term experiments (about one hour of exposure) were performed to better understand selenite transport (uptake kinetics and levels) and identify main factors influencing absorption (nutrients concentrations, pH). Long-term experiments (4 days of exposure) were performed (1) to evaluate the bioaccumulation considering environmentally relevant time scales, (2) to localize the intracellular selenium using EDAX-TEM and (3) to assess the toxicity of selenium as measured by growth impairment, ultrastructural changes, starch accumulation, and loss of pigment. Short-term experiments revealed a time-dependent linear absorption with an estimated absorbed flux of about 0.25 nmol.m -2 .nM -1 .h -1 . The absorption was proportional to ambient levels, except at very low concentrations (ca. 0.5 nM), were it was proportionally higher, suggesting that a specific but rapidly saturated transport could be used at those low concentrations. Selenite uptake was not dependent on phosphate nor carbonate concentrations. It was nevertheless inhibited by sulphate and nitrate, indicating that selenite could share common transporters with those nutrients. The accumulation was found to be maximum for intermediate pH around 7. EDAX-TEM analysis after long-term experiments revealed the presence of selenium in electron-dense granules

Sensitivity of the green algae Chlamydomonas reinhardtii to gamma radiation: Photosynthetic performance and ROS formation

International Nuclear Information System (INIS)

Gomes, Tânia; Xie, Li; Brede, Dag; Lind, Ole-Christian; Solhaug, Knut Asbjørn; Salbu, Brit

2017-01-01

Highlights: • Chlorophyll fluorescence parameters affected at higher dose rates. • Changes in PSII associated with electron transport and energy dissipation pathways. • Dose-dependent ROS production in algae exposed to gamma radiation. • Decrease in photosynthetic efficiency connected to ROS formation. - Abstract: The aquatic environment is continuously exposed to ionizing radiation from both natural and anthropogenic sources, making the characterization of ecological and health risks associated with radiation of large importance. Microalgae represent the main source of biomass production in the aquatic ecosystem, thus becoming a highly relevant biological model to assess the impacts of gamma radiation. However, little information is available on the effects of gamma radiation on microalgal species, making environmental radioprotection of this group of species challenging. In this context, the present study aimed to improve the understanding of the effects and toxic mechanisms of gamma radiation in the unicellular green algae Chlamydomonas reinhardtii focusing on the activity of the photosynthetic apparatus and ROS formation. Algal cells were exposed to gamma radiation (0.49–1677 mGy/h) for 6 h and chlorophyll fluorescence parameters obtained by PAM fluorometry, while two fluorescent probes carboxy-H 2 DFFDA and DHR 123 were used for the quantification of ROS. The alterations seen in functional parameters of C. reinhardtii PSII after 6 h of exposure to gamma radiation showed modifications of PSII energy transfer associated with electron transport and energy dissipation pathways, especially at the higher dose rates used. Results also showed that gamma radiation induced ROS in a dose-dependent manner under both light and dark conditions. The observed decrease in photosynthetic efficiency seems to be connected to the formation of ROS and can potentially lead to oxidative stress and cellular damage in chloroplasts. To our knowledge, this is the first report

New tools for chloroplast genetic engineering allow the synthesis of human growth hormone in the green alga Chlamydomonas reinhardtii.

Science.gov (United States)

Wannathong, Thanyanan; Waterhouse, Janet C; Young, Rosanna E B; Economou, Chloe K; Purton, Saul

2016-06-01

In recent years, there has been an increasing interest in the exploitation of microalgae in industrial biotechnology. Potentially, these phototrophic eukaryotes could be used for the low-cost synthesis of valuable recombinant products such as bioactive metabolites and therapeutic proteins. The algal chloroplast in particular represents an attractive target for such genetic engineering, both because it houses major metabolic pathways and because foreign genes can be targeted to specific loci within the chloroplast genome, resulting in high-level, stable expression. However, routine methods for chloroplast genetic engineering are currently available only for one species-Chlamydomonas reinhardtii-and even here, there are limitations to the existing technology, including the need for an expensive biolistic device for DNA delivery, the lack of robust expression vectors, and the undesirable use of antibiotic resistance markers. Here, we describe a new strain and vectors for targeted insertion of transgenes into a neutral chloroplast locus that (i) allow scar-less fusion of a transgenic coding sequence to the promoter/5'UTR element of the highly expressed endogenous genes psaA or atpA, (ii) employ the endogenous gene psbH as an effective but benign selectable marker, and (iii) ensure the successful integration of the transgene construct in all transformant lines. Transformation is achieved by a simple and cheap method of agitation of a DNA/cell suspension with glass beads, with selection based on the phototrophic rescue of a cell wall-deficient ΔpsbH strain. We demonstrate the utility of these tools in the creation of a transgenic line that produces high levels of functional human growth hormone.

An inorganic carbon transport system responsible for acclimation specific to air levels of CO2 in Chlamydomonas reinhardtii.

Science.gov (United States)

Wang, Yingjun; Spalding, Martin H

2006-06-27

Many photosynthetic microorganisms acclimate to CO(2) limited environments by induction and operation of CO(2)-concentrating mechanisms (CCMs). Despite their central role in CCM function, inorganic carbon (Ci) transport systems never have been identified in eukaryotic photosynthetic organisms. In the green alga Chlamydomonas reinhardtii, a mutant, pmp1, was described in 1983 with deficiencies in Ci transport, and a Pmp1 protein-associated Ci uptake system has been proposed to be responsible for Ci uptake in low CO(2) (air level)-acclimated cells. However, even though pmp1 represents the only clear genetic link to Ci transport in microalgae and is one of only a very few mutants directly affecting the CCM itself, the identity of Pmp1 has remained unknown. Physiological analyses indicate that C. reinhardtii possesses multiple Ci transport systems responsible for acclimation to different levels of limiting CO(2) and that the Pmp1-associated transport system is required specifically for low (air level) CO(2) acclimation. In the current study, we identified and characterized a pmp1 allelic mutant, air dier 1 (ad1) that, like pmp1, cannot grow in low CO(2) (350 ppm) but can grow either in high CO(2) (5% CO(2)) or in very low CO(2) (<200 ppm). Molecular analyses revealed that the Ad1/Pmp1 protein is encoded by LciB, a gene previously identified as a CO(2)-responsive gene. LciB and three related genes in C. reinhardtii compose a unique gene family that encode four closely related, apparently soluble plastid proteins with no clearly identifiable conserved motifs.

Interactive effects of copper oxide nanoparticles and light to green alga Chlamydomonas reinhardtii

Energy Technology Data Exchange (ETDEWEB)

Cheloni, Giulia; Marti, Elodie; Slaveykova, Vera I., E-mail: vera.slaveykova@unige.ch

2016-01-15

Highlights: • Comparable stability of CuO-NP suspensions under different light conditions. • UVR* inhibits growth, bleaches chlorophyll fluorescence and damages membrane. • Below 1 mg L{sup −1} CuO-NPs do not attenuate light in algal suspension. • SNL enhances significantly the effect of 0.8 mg L{sup −1} CuO-NPs on microalgae. • Synergistic interactions between UVR* and CuO-NPs. - Abstract: The present study explores the effect of light with different spectral composition on the stability of CuO-nanoparticle (CuO-NP) dispersions and their effects to green alga Chlamydomonas reinhardtii. The results showed that simulated natural light (SNL) and light with enhanced UVB radiation (UVR*) do not affect the dissolution of CuO-NPs as compared to light irradiation conditions typically used in laboratory incubator (INC). Comparable values of ζ-potential and hydrodynamic size during 24 h were found under all studied conditions. Concentrations of CuO-NPs below 1 mg L{sup −1} do not attenuate the light penetration in the algal suspensions in comparison with NP-free system. Exposure to a combination of 8 μg L{sup −1} or 0.8 mg L{sup −1} CuO-NPs and INC or SNL has no significant effect on the algal growth inhibition, algal fluorescence and membrane integrity under short-term exposure. However, an enhancement of the percentage of cells experiencing oxidative stress was observed upon exposure to 0.8 mg L{sup −1} CuO-NPs and SNL for 4 and 8 h. Combination of UVR* and 0.8 mg L{sup −1} CuO-NPs resulted in synergistic effects for all biological endpoints. Despite the photocatalytic properties of CuO-NPs no significant increase in abiotic reactive oxygen species (ROS) production under simulated solar radiation was observed suggesting that the synergistic effect observed might be correlated to other factors than CuO-NP-mediated ROS photoproduction. Tests performed with CuSO{sub 4} confirmed the important role of dissolution as toxicity driving force for lower

Optimization of the C11-BODIPY(581/591) dye for the determination of lipid oxidation in Chlamydomonas reinhardtii by flow cytometry.

Science.gov (United States)

Cheloni, Giulia; Slaveykova, Vera I

2013-10-01

Lipid oxidation is a recognized end point for the study of oxidative stress and is an important parameter to describe the mode of micropollutant action on aquatic microorganisms. Therefore, the development of quick and reliable methodologies probing the oxidative stress and damage in living cells is highly sought. In the present proof-of-concept work, we examined the potential of the fluorescent dye C11-BODIPY(591/581) to probe lipid oxidation in the green microalga Chlamydomonas reinhardtii. C11-BODIPY(591/581) staining was combined with flow cytometry measurements to obtain multiparameter information on cellular features and oxidative stress damage within single cells. First, staining conditions were optimized by exploring the capability of the dye to stain algal cells under increasing cell and dye concentrations and different staining procedures. Then lipid oxidation in algae induced by short- and long-term exposures to the three metallic micropollutants, copper, mercury, and nanoparticulate copper oxide, and the two organic contaminants, diethyldithiocarbamate (DDC) and diuron was determined. In this work we pointed out C11-BODIPY(591/581) applicability in a wide range of exposure conditions, including studies of oxidation as a function of time and that it is suitable for in vivo measurements of lipid oxidation due to its high permeation and stability in cells and its low interference with algal autofluorescence. © 2013 International Society for Advancement of Cytometry. Copyright © 2013 International Society for Advancement of Cytometry.

Proteomic analysis of a model unicellular green alga, Chlamydomonas reinhardtii, during short-term exposure to irradiance stress reveals significant down regulation of several heat-shock proteins.

Science.gov (United States)

Mahong, Bancha; Roytrakul, Suttiruk; Phaonaklop, Narumon; Wongratana, Janewit; Yokthongwattana, Kittisak

2012-03-01

Oxygenic photosynthetic organisms often suffer from excessive irradiance, which cause harmful effects to the chloroplast proteins and lipids. Photoprotection and the photosystem II repair processes are the mechanisms that plants deploy to counteract the drastic effects from irradiance stress. Although the protective and repair mechanisms seemed to be similar in most plants, many species do confer different level of tolerance toward high light. Such diversity may originate from differences at the molecular level, i.e., perception of the light stress, signal transduction and expression of stress responsive genes. Comprehensive analysis of overall changes in the total pool of proteins in an organism can be performed using a proteomic approach. In this study, we employed 2-DE/LC-MS/MS-based comparative proteomic approach to analyze total proteins of the light sensitive model unicellular green alga Chlamydomonas reinhardtii in response to excessive irradiance. Results showed that among all the differentially expressed proteins, several heat-shock proteins and molecular chaperones were surprisingly down-regulated after 3-6 h of high light exposure. Discussions were made on the possible involvement of such down regulation and the light sensitive nature of this model alga.

Sensitivity evaluation of the green alga Chlamydomonas reinhardtii to uranium by pulse amplitude modulated (PAM) fluorometry

International Nuclear Information System (INIS)

Herlory, Olivier; Bonzom, Jean-Marc; Gilbin, Rodolphe

2013-01-01

Highlights: •Our study addressed the toxicity thresholds of uranium on microalgae using PAM fluorometry. •The oxygen-evolving complex (OEC) of PSII was identified as the primary action site of uranium. •Uranium impaired the electron flux between the photosystems until almost complete inhibition. •Non-photochemical quenching was identified as the most sensitive fluorescence parameter. •PAM fluorometry provided a rapid and reasonably sensitive method for assessing stress response. -- Abstract: Although ecotoxicological studies tend to address the toxicity thresholds of uranium in freshwaters, there is a lack of information on the effects of the metal on physiological processes, particularly in aquatic plants. Knowing that uranium alters photosynthesis via impairment of the water photo-oxidation process, we determined whether pulse amplitude modulated (PAM) fluorometry was a relevant tool for assessing the impact of uranium on the green alga Chlamydomonas reinhardtii and investigated how and to what extent uranium hampered photosynthetic performance. Photosynthetic activity and quenching were assessed from fluorescence induction curves generated by PAM fluorometry, after 1 and 5 h of uranium exposure in controlled conditions. The oxygen-evolving complex (OEC) of PSII was identified as the primary action site of uranium, through alteration of the water photo-oxidation process as revealed by F 0 /F v . Limiting re-oxidation of the plastoquinone pool, uranium impaired the electron flux between the photosystems until almost complete inhibition of the PSII quantum efficiency (F ′ q /F ′ m , EC 50 = 303 ± 64 μg U L −1 after 5 h of exposure) was observed. Non-photochemical quenching (qN) was identified as the most sensitive fluorescence parameter (EC 50 = 142 ± 98 μg U L −1 after 5 h of exposure), indicating that light energy not used in photochemistry was dissipated in non-radiative processes. It was shown that parameters which stemmed from

Rubisco Activase Is Required for Optimal Photosynthesis in the Green Alga Chlamydomonas reinhardtii in a Low-CO2 Atmosphere1

Science.gov (United States)

Pollock, Steve V.; Colombo, Sergio L.; Prout, Davey L.; Godfrey, Ashley C.; Moroney, James V.

2003-01-01

This report describes a Chlamydomonas reinhardtii mutant that lacks Rubisco activase (Rca). Using the BleR (bleomycin resistance) gene as a positive selectable marker for nuclear transformation, an insertional mutagenesis screen was performed to select for cells that required a high-CO2 atmosphere for optimal growth. The DNA flanking the BleR insert of one of the high-CO2-requiring strains was cloned using thermal asymmetric interlaced-polymerase chain reaction and inverse polymerase chain reaction and sequenced. The flanking sequence matched the C. reinhardtii Rca cDNA sequence previously deposited in the National Center for Biotechnology Information database. The loss of a functional Rca in the strain was confirmed by the absence of Rca mRNA and protein. The open reading frame for Rca was cloned and expressed in pSL18, a C. reinhardtii expression vector conferring paromomycin resistance. This construct partially complemented the mutant phenotype, supporting the hypothesis that the loss of Rca was the reason the mutant grew poorly in a low-CO2 atmosphere. Sequencing of the C. reinhardtii Rca gene revealed that it contains 10 exons ranging in size from 18 to 470 bp. Low-CO2-grown rca1 cultures had a growth rate and maximum rate of photosynthesis 60% of wild-type cells. Results obtained from experiments on a cia5 rca1 double mutant also suggest that the CO2-concentrating mechanism partially compensates for the absence of an active Rca in the green alga C. reinhardtii. PMID:14605215

Cd2+ Toxicity to a Green Alga Chlamydomonas reinhardtii as Influenced by Its Adsorption on TiO2 Engineered Nanoparticles

Science.gov (United States)

Yang, Wei-Wan; Miao, Ai-Jun; Yang, Liu-Yan

2012-01-01

In the present study, Cd2+ adsorption on polyacrylate-coated TiO2 engineered nanoparticles (TiO2-ENs) and its effect on the bioavailability as well as toxicity of Cd2+ to a green alga Chlamydomonas reinhardtii were investigated. TiO2-ENs could be well dispersed in the experimental medium and their pHpzc is approximately 2. There was a quick adsorption of Cd2+ on TiO2-ENs and a steady state was reached within 30 min. A pseudo-first order kinetics was found for the time-related changes in the amount of Cd2+ complexed with TiO2-ENs. At equilibrium, Cd2+ adsorption followed the Langmuir isotherm with the maximum binding capacity 31.9, 177.1, and 242.2 mg/g when the TiO2-EN concentration was 1, 10, and 100 mg/l, respectively. On the other hand, Cd2+ toxicity was alleviated in the presence of TiO2-ENs. Algal growth was less suppressed in treatments with comparable total Cd2+ concentration but more TiO2-ENs. However, such toxicity difference disappeared and all the data points could be fitted to a single Logistic dose-response curve when cell growth inhibition was plotted against the free Cd2+ concentration. No detectable amount of TiO2-ENs was found to be associated with the algal cells. Therefore, TiO2-ENs could reduce the free Cd2+ concentration in the toxicity media, which further lowered its bioavailability and toxicity to C. reinhardtii. PMID:22403644

Origin of the polycomb repressive complex 2 and gene silencing by an E(z) homolog in the unicellular alga Chlamydomonas.

Science.gov (United States)

Shaver, Scott; Casas-Mollano, J Armando; Cerny, Ronald L; Cerutti, Heriberto

2010-05-16

Polycomb group proteins play an essential role in the maintenance of cell identity and the regulation of development in both animals and plants. The Polycomb Repressive Complex 2 (PRC2) is involved in the establishment of transcriptionally silent chromatin states, in part through its ability to methylate lysine 27 of histone H3 by the Enhancer of zeste [E(z)] subunit. The absence of PRC2 in unicellular model fungi and its function in the repression of genes vital for the development of higher eukaryotes led to the proposal that this complex may have evolved together with the emergence of multicellularity. However, we report here on the widespread presence of PRC2 core subunits in unicellular eukaryotes from the Opisthokonta, Chromalveolata and Archaeplastida supergroups. To gain insight on the role of PRC2 in single celled organisms, we characterized an E(z) homolog, EZH, in the green alga Chlamydomonas reinhardtii. RNAi-mediated suppression of EZH led to defects in the silencing of transgenes and retrotransposons as well as to a global increase in histone post-translational modifications associated with transcriptional activity, such as trimethylation of histone H3 lysine 4 and acetylation of histone H4. On the basis of the parsimony principle, our findings suggest that PRC2 appeared early in eukaryotic evolution, even perhaps in the last unicellular common ancestor of eukaryotes. One of the ancestral roles of PCR2 may have been in defense responses against intragenomic parasites such as transposable elements, prior to being co-opted for lineage specific functions like developmental regulation in multicellular eukaryotes.

NCBI nr-aa BLAST: CBRC-PHAM-01-1025 [SEVENS

Lifescience Database Archive (English)

Full Text Available CBRC-PHAM-01-1025 ref|XP_001697359.1| magnesium chelatase subunit H [Chlamydomonas ...reinhardtii] gb|EDP00299.1| magnesium chelatase subunit H [Chlamydomonas reinhardtii] XP_001697359.1 7e-05 47% ...

Genomics of Volvocine Algae

Science.gov (United States)

Umen, James G.; Olson, Bradley J.S.C.

2015-01-01

Volvocine algae are a group of chlorophytes that together comprise a unique model for evolutionary and developmental biology. The species Chlamydomonas reinhardtii and Volvox carteri represent extremes in morphological diversity within the Volvocine clade. Chlamydomonas is unicellular and reflects the ancestral state of the group, while Volvox is multicellular and has evolved numerous innovations including germ-soma differentiation, sexual dimorphism, and complex morphogenetic patterning. The Chlamydomonas genome sequence has shed light on several areas of eukaryotic cell biology, metabolism and evolution, while the Volvox genome sequence has enabled a comparison with Chlamydomonas that reveals some of the underlying changes that enabled its transition to multicellularity, but also underscores the subtlety of this transition. Many of the tools and resources are in place to further develop Volvocine algae as a model for evolutionary genomics. PMID:25883411

Alteration of proteins and pigments influence the function of photosystem I under iron deficiency from Chlamydomonas reinhardtii.

Directory of Open Access Journals (Sweden)

Venkateswarlu Yadavalli

Full Text Available BACKGROUND: Iron is an essential micronutrient for all organisms because it is a component of enzyme cofactors that catalyze redox reactions in fundamental metabolic processes. Even though iron is abundant on earth, it is often present in the insoluble ferric [Fe (III] state, leaving many surface environments Fe-limited. The haploid green alga Chlamydomonas reinhardtii is used as a model organism for studying eukaryotic photosynthesis. This study explores structural and functional changes in PSI-LHCI supercomplexes under Fe deficiency as the eukaryotic photosynthetic apparatus adapts to Fe deficiency. RESULTS: 77K emission spectra and sucrose density gradient data show that PSI and LHCI subunits are affected under iron deficiency conditions. The visible circular dichroism (CD spectra associated with strongly-coupled chlorophyll dimers increases in intensity. The change in CD signals of pigments originates from the modification of interactions between pigment molecules. Evidence from sucrose gradients and non-denaturing (green gels indicates that PSI-LHCI levels were reduced after cells were grown for 72 h in Fe-deficient medium. Ultrafast fluorescence spectroscopy suggests that red-shifted pigments in the PSI-LHCI antenna were lost during Fe stress. Further, denaturing gel electrophoresis and immunoblot analysis reveals that levels of the PSI subunits PsaC and PsaD decreased, while PsaE was completely absent after Fe stress. The light harvesting complexes were also susceptible to iron deficiency, with Lhca1 and Lhca9 showing the most dramatic decreases. These changes in the number and composition of PSI-LHCI supercomplexes may be caused by reactive oxygen species, which increase under Fe deficiency conditions. CONCLUSIONS: Fe deficiency induces rapid reduction of the levels of photosynthetic pigments due to a decrease in chlorophyll synthesis. Chlorophyll is important not only as a light-harvesting pigment, but also has a structural role

Biosorption of copper and zinc by immobilised and free algal biomass, and the effects of metal biosorption on the growth and cellular structure of Chlorella sp.and Chlamydomonas sp.isolated from rivers in Penang, Malaysia

Institute of Scientific and Technical Information of China (English)

W.O.Wan Maznah; A.T. Al-Fawwaz; Misni Surif

2012-01-01

In this study,the biosorption of copper and zinc ions by Chlorella sp.and Chlamydomonas sp.isolated from local environments in Malaysia was investigated in a batch system and by microscopic analyses.Under optimal biosorption conditions,the biosorption capacity of Chlorella sp.for copper and zinc ions was 33.4 and 28.5 mg/g,respectively,after 6 hr of biosorption in an immobilised system.Batch experiments showed that the biosorption capacity of algal biomass immobilised in the form of sodium alginate beads was higher than that of the free biomass.Scanning electron microscopy and energy-dispersive X-ray spectroscopy analyses revealed that copper and zinc were mainly sorbed at the cell surface during biosorption.Exposure to 5 mg/L of copper and zinc affected both the chlorophyll content and cell count of the algal cells after the first 12 hr of contact time.

Sensitivity evaluation of the green alga Chlamydomonas reinhardtii to uranium by pulse amplitude modulated (PAM) fluorometry

Energy Technology Data Exchange (ETDEWEB)

Herlory, Olivier, E-mail: olivier.herlory@gmail.com [IRSN-Laboratoire d’Ecotoxicologie des Radionucléides, Centre de Cadarache, BP3, 13115 Saint Paul lez Durance (France); Bonzom, Jean-Marc, E-mail: jean-marc.bonzom@irsn.fr [IRSN-Laboratoire d’Ecotoxicologie des Radionucléides, Centre de Cadarache, BP3, 13115 Saint Paul lez Durance (France); Gilbin, Rodolphe, E-mail: rodolphe.gilbin@irsn.fr [IRSN-Laboratoire de Biogéochimie, Biodisponibilité et Transferts des Radionucléides, Centre de Cadarache, BP3, 13115 Saint Paul lez Durance (France)

2013-09-15

Highlights: •Our study addressed the toxicity thresholds of uranium on microalgae using PAM fluorometry. •The oxygen-evolving complex (OEC) of PSII was identified as the primary action site of uranium. •Uranium impaired the electron flux between the photosystems until almost complete inhibition. •Non-photochemical quenching was identified as the most sensitive fluorescence parameter. •PAM fluorometry provided a rapid and reasonably sensitive method for assessing stress response. -- Abstract: Although ecotoxicological studies tend to address the toxicity thresholds of uranium in freshwaters, there is a lack of information on the effects of the metal on physiological processes, particularly in aquatic plants. Knowing that uranium alters photosynthesis via impairment of the water photo-oxidation process, we determined whether pulse amplitude modulated (PAM) fluorometry was a relevant tool for assessing the impact of uranium on the green alga Chlamydomonas reinhardtii and investigated how and to what extent uranium hampered photosynthetic performance. Photosynthetic activity and quenching were assessed from fluorescence induction curves generated by PAM fluorometry, after 1 and 5 h of uranium exposure in controlled conditions. The oxygen-evolving complex (OEC) of PSII was identified as the primary action site of uranium, through alteration of the water photo-oxidation process as revealed by F{sub 0}/F{sub v}. Limiting re-oxidation of the plastoquinone pool, uranium impaired the electron flux between the photosystems until almost complete inhibition of the PSII quantum efficiency (F{sup ′}{sub q}/F{sup ′}{sub m}, EC{sub 50} = 303 ± 64 μg U L{sup −1} after 5 h of exposure) was observed. Non-photochemical quenching (qN) was identified as the most sensitive fluorescence parameter (EC{sub 50} = 142 ± 98 μg U L{sup −1} after 5 h of exposure), indicating that light energy not used in photochemistry was dissipated in non-radiative processes. It was shown

Real-time monitoring of genetically modified Chlamydomonas reinhardtii during the Foton M3 space mission

Science.gov (United States)

Lambreva, M.; Rea, G.; Antonacci, A.; Serafini, A.; Damasso, M.; Pastorelli, S.; Margonelli, A.; Johanningmeier, U.; Bertalan, I.; Pezzotti, G.; Giardi, M. T.

2008-09-01

Long-term space exploration, colonization or habitation requires biological life support systems capable to cope with the deleterious space environment. The use of oxygenic photosynthetic microrganisms is an intriguing possibility mainly for food, O2 and nutraceutical compounds production. The critical points of utilizing plants- or algae-based life support systems are the microgravity and the ionizing radiation, which can influence the performance of these organisms. The aim of the present study was to assess the effects of space environment on the photosynthetic activity of various microrganisms and to select space stresstolerant strains. Photosystem II D1 protein sitedirected and random mutants of the unicellular green alga Chlamydomonas reinhardtii [1] were used as a model system to test and select the amino acid substitutions capable to account for space stress tolerance. We focussed our studies also on the accumulation of the Photosystem II photoprotective carotenoids (the xantophylls violaxanthin, anteraxanthin and zeaxanthin), powerful antioxidants that epidemiological studies demonstrated to be human vision protectors. For this purpose some mutants modified at the level of enzymes involved in the biosynthesis of xanthophylls were included in the study [2]. To identify the consequences of the space environment on the photosynthetic apparatus the changes in the Photosystem II efficiency were monitored in real time during the ESA-Russian Foton- M3 mission in September 2007. For the space flight a high-tech, multicell fluorescence detector, Photo-II, was designed and built by the Centre for Advanced Research in Space Optics in collaboration with Kayser-Italy, Biosensor and DAS. Photo-II is an automatic device developed to measure the chlorophyll fluorescence and to provide a living conditions for several different algae strains (Fig.1). Twelve different C. reinhardti strains were analytically selected and two replications for each strain were brought to space

Buoyant triacylglycerol-filled green algae and methods therefor

Science.gov (United States)

Goodenough, Ursula; Goodson, Carrie

2015-04-14

Cultures of Chlamydomonas are disclosed comprising greater than 340 mg/l triacylglycerols (TAG). The cultures can include buoyant Chlamydomonas. Methods of forming the cultures are also disclosed. In some embodiments, these methods comprise providing Chlamydomonas growing in log phase in a first culture medium comprising a nitrogen source and acetate, replacing the first culture medium with a second medium comprising acetate but no nitrogen source, and subsequently supplementing the second medium with additional acetate. In some embodiments, a culture can comprise at least 1,300 mg/l triacyglycerols. In some embodiments, cultures can be used to produce a biofuel such as biodiesel.

Multiple stressor effects in Chlamydomonas reinhardtii – Toward understanding mechanisms of interaction between effects of ultraviolet radiation and chemical pollutants

Energy Technology Data Exchange (ETDEWEB)

Korkaric, Muris [Eawag, Swiss Federal Institute of Aquatic Science and Technology, Department of Environmental Toxicology, 8600, Duebendorf (Switzerland); ETH Zürich, Institute of Biogeochemistry and Pollutant Dynamics, 8092 Zürich (Switzerland); Behra, Renata; Fischer, Beat B. [Eawag, Swiss Federal Institute of Aquatic Science and Technology, Department of Environmental Toxicology, 8600, Duebendorf (Switzerland); Junghans, Marion [Swiss Center for Applied Ecotoxicology Eawag-EPFL, 8600, Duebendorf (Switzerland); Eggen, Rik I.L., E-mail: rik.eggen@eawag.ch [Eawag, Swiss Federal Institute of Aquatic Science and Technology, Department of Environmental Toxicology, 8600, Duebendorf (Switzerland); ETH Zürich, Institute of Biogeochemistry and Pollutant Dynamics, 8092 Zürich (Switzerland)

2015-05-15

Highlights: • Systematic study of multiple stressor effects of UVR and chemicals in C. reinhardtii. • UVR and chemicals did not act independently on algal photosynthesis and reproduction. • Multiple stressor effects of UVR and chemicals depended on chemical MOA. • Synergistic effect interactions not limited to oxidative stress inducing chemicals. • Multiple MOAs of UVR may limit applicability of current prediction models. - Abstract: The effects of chemical pollutants and environmental stressors, such as ultraviolet radiation (UVR), can interact when organisms are simultaneously exposed, resulting in higher (synergistic) or lower (antagonistic) multiple stressor effects than expected based on the effects of single stressors. Current understanding of interactive effects is limited due to a lack of mechanism-based multiple stressor studies. It has been hypothesized that effect interactions may generally occur if chemical and non-chemical stressors cause similar physiological effects in the organism. To test this hypothesis, we exposed the model green alga Chlamydomonas reinhardtii to combinations of UVR and single chemicals displaying modes of action (MOA) similar or dissimilar to the impact of UVR on photosynthesis. Stressor interactions were analyzed based on the independent action model. Effect interactions were found to depend on the MOA of the chemicals, and also on their concentrations, the exposure time and the measured endpoint. Indeed, only chemicals assumed to cause effects on photosynthesis similar to UVR showed interactions with UVR on photosynthetic yield: synergistic in case of Cd(II) and paraquat and antagonistic in case of diuron. No interaction on photosynthesis was observed for S-metolachlor, which acts dissimilarly to UVR. However, combined effects of S-metolachlor and UVR on algal reproduction were synergistic, highlighting the importance of considering additional MOA of UVR. Possible mechanisms of stressor effect interactions are

Multiple stressor effects in Chlamydomonas reinhardtii – Toward understanding mechanisms of interaction between effects of ultraviolet radiation and chemical pollutants

International Nuclear Information System (INIS)

Korkaric, Muris; Behra, Renata; Fischer, Beat B.; Junghans, Marion; Eggen, Rik I.L.

2015-01-01

Highlights: • Systematic study of multiple stressor effects of UVR and chemicals in C. reinhardtii. • UVR and chemicals did not act independently on algal photosynthesis and reproduction. • Multiple stressor effects of UVR and chemicals depended on chemical MOA. • Synergistic effect interactions not limited to oxidative stress inducing chemicals. • Multiple MOAs of UVR may limit applicability of current prediction models. - Abstract: The effects of chemical pollutants and environmental stressors, such as ultraviolet radiation (UVR), can interact when organisms are simultaneously exposed, resulting in higher (synergistic) or lower (antagonistic) multiple stressor effects than expected based on the effects of single stressors. Current understanding of interactive effects is limited due to a lack of mechanism-based multiple stressor studies. It has been hypothesized that effect interactions may generally occur if chemical and non-chemical stressors cause similar physiological effects in the organism. To test this hypothesis, we exposed the model green alga Chlamydomonas reinhardtii to combinations of UVR and single chemicals displaying modes of action (MOA) similar or dissimilar to the impact of UVR on photosynthesis. Stressor interactions were analyzed based on the independent action model. Effect interactions were found to depend on the MOA of the chemicals, and also on their concentrations, the exposure time and the measured endpoint. Indeed, only chemicals assumed to cause effects on photosynthesis similar to UVR showed interactions with UVR on photosynthetic yield: synergistic in case of Cd(II) and paraquat and antagonistic in case of diuron. No interaction on photosynthesis was observed for S-metolachlor, which acts dissimilarly to UVR. However, combined effects of S-metolachlor and UVR on algal reproduction were synergistic, highlighting the importance of considering additional MOA of UVR. Possible mechanisms of stressor effect interactions are

Robust expression and secretion of Xylanase1 in Chlamydomonas reinhardtii by fusion to a selection gene and processing with the FMDV 2A peptide.

Directory of Open Access Journals (Sweden)

Beth A Rasala

Full Text Available Microalgae have recently received attention as a potential low-cost host for the production of recombinant proteins and novel metabolites. However, a major obstacle to the development of algae as an industrial platform has been the poor expression of heterologous genes from the nuclear genome. Here we describe a nuclear expression strategy using the foot-and-mouth-disease-virus 2A self-cleavage peptide to transcriptionally fuse heterologous gene expression to antibiotic resistance in Chlamydomonas reinhardtii. We demonstrate that strains transformed with ble-2A-GFP are zeocin-resistant and accumulate high levels of GFP that is properly 'cleaved' at the FMDV 2A peptide resulting in monomeric, cytosolic GFP that is easily detectable by in-gel fluorescence analysis or fluorescent microscopy. Furthermore, we used our ble2A nuclear expression vector to engineer the heterologous expression of the industrial enzyme, xylanase. We demonstrate that linking xyn1 expression to ble2A expression on the same open reading frame led to a dramatic (~100-fold increase in xylanase activity in cells lysates compared to the unlinked construct. Finally, by inserting an endogenous secretion signal between the ble2A and xyn1 coding regions, we were able to target monomeric xylanase for secretion. The novel microalgae nuclear expression strategy described here enables the selection of transgenic lines that are efficiently expressing the heterologous gene-of-interest and should prove valuable for basic research as well as algal biotechnology.

Bioaccumulation and subcellular partitioning of Cr(III) and Cr(VI) in the freshwater green alga Chlamydomonas reinhardtii

Energy Technology Data Exchange (ETDEWEB)

Aharchaou, Imad [Laboratoire Interdisciplinaire des Environnements Continentaux, UMR 7360, Université de Lorraine and CNRS, 8 rue du Général Delestraint, 57070 Metz (France); Rosabal, Maikel; Liu, Fengjie [Institut National de la Recherche Scientifique, Centre Eau Terre Environnement (INRS-ETE), 490 rue de la Couronne, Québec (Québec) G1K 9A9 (Canada); Battaglia, Eric; Vignati, Davide A.L. [Laboratoire Interdisciplinaire des Environnements Continentaux, UMR 7360, Université de Lorraine and CNRS, 8 rue du Général Delestraint, 57070 Metz (France); Fortin, Claude, E-mail: claude.fortin@ete.inrs.ca [Institut National de la Recherche Scientifique, Centre Eau Terre Environnement (INRS-ETE), 490 rue de la Couronne, Québec (Québec) G1K 9A9 (Canada)

2017-01-15

Highlights: • C. reinhardtii accumulated similar levels of Cr(III) and Cr(VI). • The subcellular partitioning of Cr(III) and Cr(VI) was similar. • Cr(III) and Cr(VI) associated mainly with organelles and heat-stable proteins. • Metallomic analysis showed two main Cr-binding biomolecules after 72 h of exposure. - Abstract: Chromium occurs in aquatic environments under two main redox forms, namely Cr(III) and Cr(VI), with different geochemical and biochemical properties. Cr(VI) readily crosses biological membranes of living organisms and once inside the cells it undergoes a rapid reduction to Cr(III). The route of entry for the latter form is, however, poorly known. Using the radioactive tracer {sup 51}Cr we compared the accumulation (absorption and adsorption) of the two Cr forms by the green unicellular alga Chlamydomonas reinhardii after 1 h and 72 h of exposure to 100 nM of either Cr(III) or Cr(VI) at pH 7. Both Cr forms had similar accumulation, with a major part in the extracellular (adsorbed) fraction after 1 h and a major part of total accumulated Cr in the intracellular (absorbed) fraction after 72 h. We also investigated the intracellular partitioning of Cr using an operational fractionation scheme and found that both Cr forms had similar distributions among fractions: Cr was mostly associated with organelles (23 ± 12% after 1 h and 37 ± 7% after 72 h) and cytosolic heat-stable proteins and peptides (39 ± 18% after 1 h and 35 ± 3% after 72 h) fractions. Further investigations using a metallomic approach (SEC-ICP-MS) were performed with the heat-stable proteins and peptides fraction to compare the distribution of the two Cr forms among various biomolecules of this fraction. One Cr-binding biomolecule (∼28 kDa) appeared after 1 h of exposure for both Cr species. After 72 h another biomolecule of lower molecular weight (∼0.7 kDa) was involved in binding Cr and higher signal intensities were observed for Cr(VI) than for Cr(III). We show, for the

Bioaccumulation and subcellular partitioning of Cr(III) and Cr(VI) in the freshwater green alga Chlamydomonas reinhardtii

International Nuclear Information System (INIS)

Aharchaou, Imad; Rosabal, Maikel; Liu, Fengjie; Battaglia, Eric; Vignati, Davide A.L.; Fortin, Claude

2017-01-01

Highlights: • C. reinhardtii accumulated similar levels of Cr(III) and Cr(VI). • The subcellular partitioning of Cr(III) and Cr(VI) was similar. • Cr(III) and Cr(VI) associated mainly with organelles and heat-stable proteins. • Metallomic analysis showed two main Cr-binding biomolecules after 72 h of exposure. - Abstract: Chromium occurs in aquatic environments under two main redox forms, namely Cr(III) and Cr(VI), with different geochemical and biochemical properties. Cr(VI) readily crosses biological membranes of living organisms and once inside the cells it undergoes a rapid reduction to Cr(III). The route of entry for the latter form is, however, poorly known. Using the radioactive tracer "5"1Cr we compared the accumulation (absorption and adsorption) of the two Cr forms by the green unicellular alga Chlamydomonas reinhardii after 1 h and 72 h of exposure to 100 nM of either Cr(III) or Cr(VI) at pH 7. Both Cr forms had similar accumulation, with a major part in the extracellular (adsorbed) fraction after 1 h and a major part of total accumulated Cr in the intracellular (absorbed) fraction after 72 h. We also investigated the intracellular partitioning of Cr using an operational fractionation scheme and found that both Cr forms had similar distributions among fractions: Cr was mostly associated with organelles (23 ± 12% after 1 h and 37 ± 7% after 72 h) and cytosolic heat-stable proteins and peptides (39 ± 18% after 1 h and 35 ± 3% after 72 h) fractions. Further investigations using a metallomic approach (SEC-ICP-MS) were performed with the heat-stable proteins and peptides fraction to compare the distribution of the two Cr forms among various biomolecules of this fraction. One Cr-binding biomolecule (∼28 kDa) appeared after 1 h of exposure for both Cr species. After 72 h another biomolecule of lower molecular weight (∼0.7 kDa) was involved in binding Cr and higher signal intensities were observed for Cr(VI) than for Cr(III). We show, for the

Carbon allocation and element composition in four Chlamydomonas mutants defective in genes related to the CO2 concentrating mechanism.

Science.gov (United States)

Memmola, Francesco; Mukherjee, Bratati; Moroney, James V; Giordano, Mario

2014-09-01

Four mutants of Chlamydomonas reinhardtii with defects in different components of the CO2 concentrating mechanism (CCM) or in Rubisco activase were grown autotrophically at high pCO2 and then transferred to low pCO2, in order to study the role of different components of the CCM on carbon allocation and elemental composition. To study carbon allocation, we measured the relative size of the main organic pools by Fourier Transform Infrared spectroscopy. Total reflection X-ray fluorescence was used to analyze the elemental composition of algal cells. Our data show that although the organic pools increased their size at high CO2 in all strains, their stoichiometry was highly homeostatic, i.e., the ratios between carbohydrates and proteins, lipid and proteins, and carbohydrates and lipids, did not change significantly. The only exception was the wild-type 137c, in which proteins decreased relative to carbohydrates and lipids, when the cells were transferred to low CO2. It is noticeable that the two wild types used in this study responded differently to the transition from high to low CO2. Malfunctions of the CCM influenced the concentration of several elements, somewhat altering cell elemental stoichiometry: especially the C/P and N/P ratios changed appreciably in almost all strains as a function of the growth CO2 concentration, except in 137c and the Rubisco activase mutant rca1. In strain cia3, defective in the lumenal carbonic anhydrase (CA), the cell quotas of P, S, Ca, Mn, Fe, and Zn were about 5-fold higher at low CO2 than at high CO2. A Principle Components Analysis showed that, mostly because of its elemental composition, cia3 behaved in a substantially different way from all other strains, at low CO2. The lumenal CA thus plays a crucial role, not only for the correct functioning of the CCM, but also for element utilization. Not surprisingly, growth at high CO2 attenuated differences among strains.

RNAi knock-down of LHCBM1, 2 and 3 increases photosynthetic H2 production efficiency of the green alga Chlamydomonas reinhardtii.

Directory of Open Access Journals (Sweden)

Melanie Oey

Full Text Available Single cell green algae (microalgae are rapidly emerging as a platform for the production of sustainable fuels. Solar-driven H2 production from H2O theoretically provides the highest-efficiency route to fuel production in microalgae. This is because the H2-producing hydrogenase (HYDA is directly coupled to the photosynthetic electron transport chain, thereby eliminating downstream energetic losses associated with the synthesis of carbohydrate and oils (feedstocks for methane, ethanol and oil-based fuels. Here we report the simultaneous knock-down of three light-harvesting complex proteins (LHCMB1, 2 and 3 in the high H2-producing Chlamydomonas reinhardtii mutant Stm6Glc4 using an RNAi triple knock-down strategy. The resultant Stm6Glc4L01 mutant exhibited a light green phenotype, reduced expression of LHCBM1 (20.6% ±0.27%, LHCBM2 (81.2% ±0.037% and LHCBM3 (41.4% ±0.05% compared to 100% control levels, and improved light to H2 (180% and biomass (165% conversion efficiencies. The improved H2 production efficiency was achieved at increased solar flux densities (450 instead of ∼100 µE m(-2 s(-1 and high cell densities which are best suited for microalgae production as light is ideally the limiting factor. Our data suggests that the overall improved photon-to-H2 conversion efficiency is due to: 1 reduced loss of absorbed energy by non-photochemical quenching (fluorescence and heat losses near the photobioreactor surface; 2 improved light distribution in the reactor; 3 reduced photoinhibition; 4 early onset of HYDA expression and 5 reduction of O2-induced inhibition of HYDA. The Stm6Glc4L01 phenotype therefore provides important insights for the development of high-efficiency photobiological H2 production systems.

Submicron and nano formulations of titanium dioxide and zinc oxide stimulate unique cellular toxicological responses in the green microalga Chlamydomonas reinhardtii

Energy Technology Data Exchange (ETDEWEB)

Gunawan, Cindy, E-mail: c.gunawan@unsw.edu.au [ARC Centre of Excellence for Functional Nanomaterials, School of Chemical Engineering, The University of New South Wales, Sydney, NSW (Australia); Sirimanoonphan, Aunchisa [ARC Centre of Excellence for Functional Nanomaterials, School of Chemical Engineering, The University of New South Wales, Sydney, NSW (Australia); Teoh, Wey Yang [Clean Energy and Nanotechnology (CLEAN) Laboratory, School of Energy and Environment, City University of Hong Kong, Kowloon, Hong Kong Special Administrative Region (Hong Kong); Marquis, Christopher P., E-mail: c.marquis@unsw.edu.au [School of Biotechnology and Biomolecular Sciences, The University of New South Wales, Sydney, NSW (Australia); Amal, Rose [ARC Centre of Excellence for Functional Nanomaterials, School of Chemical Engineering, The University of New South Wales, Sydney, NSW (Australia)

2013-09-15

Highlights: • Uptake of TiO{sub 2} solids by C. reinhardtii generates ROS as an early stress response. • Submicron and nanoTiO{sub 2} exhibit benign effect on cell proliferation. • Uptake of ZnO solids and leached zinc by C. reinhardtii inhibit the alga growth. • No cellular oxidative stress is detected with submicron and nano ZnO exposure. • The toxicity of particles is not necessarily mediated by cellular oxidative stress. -- Abstract: The work investigates the eco-cytoxicity of submicron and nano TiO{sub 2} and ZnO, arising from the unique interactions of freshwater microalga Chlamydomonas reinhardtii to soluble and undissolved components of the metal oxides. In a freshwater medium, submicron and nano TiO{sub 2} exist as suspended aggregates with no-observable leaching. Submicron and nano ZnO undergo comparable concentration-dependent fractional leaching, and exist as dissolved zinc and aggregates of undissolved ZnO. Cellular internalisation of solid TiO{sub 2} stimulates cellular ROS generation as an early stress response. The cellular redox imbalance was observed for both submicron and nano TiO{sub 2} exposure, despite exhibiting benign effects on the alga proliferation (8-day EC50 > 100 mg TiO{sub 2}/L). Parallel exposure of C. reinhardtii to submicron and nano ZnO saw cellular uptake of both the leached zinc and solid ZnO and resulting in inhibition of the alga growth (8-day EC50 ≥ 0.01 mg ZnO/L). Despite the sensitivity, no zinc-induced cellular ROS generation was detected, even at 100 mg ZnO/L exposure. Taken together, the observations confront the generally accepted paradigm of cellular oxidative stress-mediated cytotoxicity of particles. The knowledge of speciation of particles and the corresponding stimulation of unique cellular responses and cytotoxicity is vital for assessment of the environmental implications of these materials.

Submicron and nano formulations of titanium dioxide and zinc oxide stimulate unique cellular toxicological responses in the green microalga Chlamydomonas reinhardtii

International Nuclear Information System (INIS)

Gunawan, Cindy; Sirimanoonphan, Aunchisa; Teoh, Wey Yang; Marquis, Christopher P.; Amal, Rose

2013-01-01

Highlights: • Uptake of TiO 2 solids by C. reinhardtii generates ROS as an early stress response. • Submicron and nanoTiO 2 exhibit benign effect on cell proliferation. • Uptake of ZnO solids and leached zinc by C. reinhardtii inhibit the alga growth. • No cellular oxidative stress is detected with submicron and nano ZnO exposure. • The toxicity of particles is not necessarily mediated by cellular oxidative stress. -- Abstract: The work investigates the eco-cytoxicity of submicron and nano TiO 2 and ZnO, arising from the unique interactions of freshwater microalga Chlamydomonas reinhardtii to soluble and undissolved components of the metal oxides. In a freshwater medium, submicron and nano TiO 2 exist as suspended aggregates with no-observable leaching. Submicron and nano ZnO undergo comparable concentration-dependent fractional leaching, and exist as dissolved zinc and aggregates of undissolved ZnO. Cellular internalisation of solid TiO 2 stimulates cellular ROS generation as an early stress response. The cellular redox imbalance was observed for both submicron and nano TiO 2 exposure, despite exhibiting benign effects on the alga proliferation (8-day EC50 > 100 mg TiO 2 /L). Parallel exposure of C. reinhardtii to submicron and nano ZnO saw cellular uptake of both the leached zinc and solid ZnO and resulting in inhibition of the alga growth (8-day EC50 ≥ 0.01 mg ZnO/L). Despite the sensitivity, no zinc-induced cellular ROS generation was detected, even at 100 mg ZnO/L exposure. Taken together, the observations confront the generally accepted paradigm of cellular oxidative stress-mediated cytotoxicity of particles. The knowledge of speciation of particles and the corresponding stimulation of unique cellular responses and cytotoxicity is vital for assessment of the environmental implications of these materials

Variable selectivity and the role of nutritional quality in food selection by a planktonic rotifer

International Nuclear Information System (INIS)

Sierszen, M.E.

1990-01-01

To investigate the potential for selective feeding to enhance fitness, I test the hypothesis that an herbivorous zooplankter selects those food items that best support its reproduction. Under this hypothesis, growth and reproduction on selected food items should be higher than on less preferred items. The hypothesis is not supported. In situ selectivity by the rotifer Keratella taurocephala for Cryptomonas relative to Chlamydomonas goes through a seasonal cycle, in apparent response to fluctuating Cryptomonas populations. However, reproduction on a unialgal diet of Cryptomonas is consistently high and similar to that on Chlamydomonas. Oocystis, which also supports reproduction equivalent to that supported by Chlamydomonas, is sometimes rejected by K. taurocephala. In addition, K. taurocephala does not discriminate between Merismopedia and Chlamydomonas even though Merismopedia supports virtually no reproduction by the rotifer. Selection by K. taurocephala does not simply maximize the intake of food items that yield high reproduction. Selectivity is a complex, dynamic process, one function of which may be the exploitation of locally or seasonally abundant foods. (author)

Linoleic Acid-Induced Ultra-Weak Photon Emission from Chlamydomonas reinhardtii as a Tool for Monitoring of Lipid Peroxidation in the Cell Membranes

Science.gov (United States)

Prasad, Ankush; Pospíšil, Pavel

2011-01-01

Reactive oxygen species formed as a response to various abiotic and biotic stresses cause an oxidative damage of cellular component such are lipids, proteins and nucleic acids. Lipid peroxidation is considered as one of the major processes responsible for the oxidative damage of the polyunsaturated fatty acid in the cell membranes. Various methods such as a loss of polyunsaturated fatty acids, amount of the primary and the secondary products are used to monitor the level of lipid peroxidation. To investigate the use of ultra-weak photon emission as a non-invasive tool for monitoring of lipid peroxidation, the involvement of lipid peroxidation in ultra-weak photon emission was studied in the unicellular green alga Chlamydomonas reinhardtii. Lipid peroxidation initiated by addition of exogenous linoleic acid to the cells was monitored by ultra-weak photon emission measured with the employment of highly sensitive charged couple device camera and photomultiplier tube. It was found that the addition of linoleic acid to the cells significantly increased the ultra-weak photon emission that correlates with the accumulation of lipid peroxidation product as measured using thiobarbituric acid assay. Scavenging of hydroxyl radical by mannitol, inhibition of intrinsic lipoxygenase by catechol and removal of molecular oxygen considerably suppressed ultra-weak photon emission measured after the addition of linoleic acid. The photon emission dominated at the red region of the spectrum with emission maximum at 680 nm. These observations reveal that the oxidation of linoleic acid by hydroxyl radical and intrinsic lipoxygenase results in the ultra-weak photon emission. Electronically excited species such as excited triplet carbonyls are the likely candidates for the primary excited species formed during the lipid peroxidation, whereas chlorophylls are the final emitters of photons. We propose here that the ultra-weak photon emission can be used as a non-invasive tool for the

Flow Cytometry Pulse Width Data Enables Rapid and Sensitive Estimation of Biomass Dry Weight in the Microalgae Chlamydomonas reinhardtii and Chlorella vulgaris

Science.gov (United States)

Chioccioli, Maurizio; Hankamer, Ben; Ross, Ian L.

2014-01-01

Dry weight biomass is an important parameter in algaculture. Direct measurement requires weighing milligram quantities of dried biomass, which is problematic for small volume systems containing few cells, such as laboratory studies and high throughput assays in microwell plates. In these cases indirect methods must be used, inducing measurement artefacts which vary in severity with the cell type and conditions employed. Here, we utilise flow cytometry pulse width data for the estimation of cell density and biomass, using Chlorella vulgaris and Chlamydomonas reinhardtii as model algae and compare it to optical density methods. Measurement of cell concentration by flow cytometry was shown to be more sensitive than optical density at 750 nm (OD750) for monitoring culture growth. However, neither cell concentration nor optical density correlates well to biomass when growth conditions vary. Compared to the growth of C. vulgaris in TAP (tris-acetate-phosphate) medium, cells grown in TAP + glucose displayed a slowed cell division rate and a 2-fold increased dry biomass accumulation compared to growth without glucose. This was accompanied by increased cellular volume. Laser scattering characteristics during flow cytometry were used to estimate cell diameters and it was shown that an empirical but nonlinear relationship could be shown between flow cytometric pulse width and dry weight biomass per cell. This relationship could be linearised by the use of hypertonic conditions (1 M NaCl) to dehydrate the cells, as shown by density gradient centrifugation. Flow cytometry for biomass estimation is easy to perform, sensitive and offers more comprehensive information than optical density measurements. In addition, periodic flow cytometry measurements can be used to calibrate OD750 measurements for both convenience and accuracy. This approach is particularly useful for small samples and where cellular characteristics, especially cell size, are expected to vary during growth. PMID

Selenite -Se(4)- uptake mechanisms in the unicellular green alga Chlamydomonas reinhardtii: bioaccumulation and effects induced on growth and ultrastructure

International Nuclear Information System (INIS)

Morlon, H.

2005-03-01

Selenium is an essential element, but becomes very toxic at higher concentrations. It occurs in the environment at concentrations ranging from nM to μM and selenium pollution is a worldwide phenomenon. This works aims at improving the knowledge on the interactions between selenite - Se(IV) - and a freshwater phyto-planktonic organism: the unicellular green algae Chlamydomonas reinhardtii. The aim of the performed experiments were: i) to investigate selenite -Se(IV)- uptake mechanisms in C. reinhardtii, using Se 75 as a tracer in short term exposures ( -2 .nM -1 .h -1 . The uptake was proportional to ambient levels in a broad range of intermediate concentrations (from nM to μM). However, fluxes were higher at very low concentrations ( μM), suggesting that a high affinity but rapidly saturated transport mechanism could be used at low concentrations, in parallel with a low affinity mechanism that would only saturate at high concentrations (∼mM). The latter could involve transporters used by sulphate and nitrates, as suggested by the inhibition of selenite uptake by those element. Se(IV) speciation changes with pH did not induce significant effect on bioavailability. On the basis of the relationship between Se concentration and maximal cell density achieved, an EC50 of 80 μM ([64; 98]) was derived. No adaptation mechanism were observed as the same the same toxicity was quantified for Se-pre-exposed algae. Observations by TEM suggested chloroplasts as the first target of selenite cytotoxicity, with effects on the stroma, thylakoids and pyrenoids. At higher concentrations, we could observe an increase in the number and volume of starch grains. For the cell collected at 96 h, electron-dense granules were observed. Energy-dispersive X-ray microanalysis revealed that they contained selenium and were also rich in calcium and phosphorus. Finally, growth inhibition was highly correlated to the bioaccumulation of selenite. The latter was inhibited by increasing

Toxicity of selenite in the unicellular green alga Chlamydomonas reinhardtii: Comparison between effects at the population and sub-cellular level

International Nuclear Information System (INIS)

Morlon, Helene; Fortin, Claude; Floriani, Magali; Adam, Christelle; Garnier-Laplace, Jacqueline; Boudou, Alain

2005-01-01

The toxicity of selenium in aquatic ecosystems is mainly linked to its uptake and biotransformation by micro-organisms, and its subsequent transfer upwards into the food chain. Thus, organisms at low trophic level, such as algae, play a crucial role. The aim of our study was to investigate the biological effects of selenite on Chlamydomonas reinhardtii, both at the sub-cellular level (effect on ultrastructure) and at the population level (effect on growth). The cells were grown under batch culture conditions in well-defined media and exposed to waterborne selenite at concentrations up to 500 μM; i.e. up to lethal conditions. Based on the relationship between Se concentration and cell density achieved after a 96 h exposure period, an EC 50 of 80 μM with a 95% confidence interval ranging between 64 and 98 μM was derived. No adaptation mechanisms were observed: the same toxicity was quantified for algae pre-contaminated with Se. The inhibition of growth was linked to impairments observed at the sub-cellular level. The intensity of the ultrastructural damages caused by selenite exposure depended on the level and duration of exposure. Observations by TEM suggested chloroplasts as the first target of selenite cytotoxicity, with effects on the stroma, thylakoids and pyrenoids. At higher concentrations, we could observe an increase in the number and volume of starch grains. For cells collected at 96 h, electron-dense granules were observed. Energy-dispersive X-ray microanalysis revealed that these granules contained selenium and were also rich in calcium and phosphorus. This study confirms that the direct toxicity of selenite on the phytoplankton biomass is not likely to take place at concentrations found in the environment. At higher concentrations, the link between effects at the sub-cellular and population levels, the over-accumulation of starch, and the formation of dense granules containing selenium are reported for the first time in the literature for a

The organization structure and regulatory elements of Chlamydomonas histone genes reveal features linking plant and animal genes.

Science.gov (United States)

Fabry, S; Müller, K; Lindauer, A; Park, P B; Cornelius, T; Schmitt, R

1995-09-01

The genome of the green alga Chlamydomonas reinhardtii contains approximately 15 gene clusters of the nucleosomal (or core) histone H2A, H2B, H3 and H4 genes and at least one histone H1 gene. Seven non-allelic histone gene loci were isolated from a genomic library, physically mapped, and the nucleotide sequences of three isotypes of each core histone gene species and one linked H1 gene determined. The core histone genes are organized in clusters of H2A-H2B and H3-H4 pairs, in which each gene pair shows outwardly divergent transcription from a short (< 300 bp) intercistronic region. These intercistronic regions contain typically conserved promoter elements, namely a TATA-box and the three motifs TGGCCAG-G(G/C)-CGAG, CGTTGACC and CGGTTG. Different from the genes of higher plants, but like those of animals and the related alga Volvox, the 3' untranslated regions contain no poly A signal, but a palindromic sequence (3' palindrome) essential for mRNA processing is present. One single H1 gene was found in close linkage to a H2A-H2B pair. The H1 upstream region contains the octameric promoter element GGTTGACC (also found upstream of the core histone genes) and two specific sequence motifs that are shared only with the Volvox H1 promoters. This suggests differential transcription of the H1 and the core histone genes. The H1 gene is interrupted by two introns. Unlike Volvox H3 genes, the three sequenced H3 isoforms are intron-free. Primer-directed PCR of genomic DNA demonstrated, however, that at least 8 of the about 15 H3 genes do contain one intron at a conserved position. In synchronized C. reinhardtii cells, H4 mRNA levels (representative of all core histone mRNAs) peak during cell division, suggesting strict replication-dependent gene control. The derived peptide sequences place C. reinhardtii core histones closer to plants than to animals, except that the H2A histones are more animal-like. The peptide sequence of histone H1 is closely related to the V. carteri VH1-II

Construction of Marker-Free Transgenic Strains of Chlamydomonas reinhardtii Using a Cre/loxP-Mediated Recombinase System.

Science.gov (United States)

Kasai, Yuki; Harayama, Shigeaki

2016-01-01

The Escherichia coli bacteriophage P1 encodes a site-specific recombinase called Cre and two 34-bp target sites of Cre recombinase called loxP. The Cre/loxP system has been used to achieve targeted insertion and precise deletion in many animal and plant genomes. The Cre/loxP system has particularly been used for the removal of selectable marker genes to create marker-free transgenic organisms. For the first time, we applied the Cre/loxP-mediated site-specific recombination system to Chlamydomonas reinhardtii to construct marker-free transgenic strains. Specifically, C. reinhardtii strains cc4350 and cc124 carrying an aphVIII expression cassette flanked by two direct repeats of loxP were constructed. Separately, a synthetic Cre recombinase gene (CrCRE), the codons of which were optimized for expression in C. reinhardtii, was synthesized, and a CrCRE expression cassette was introduced into strain cc4350 carrying a single copy of the loxP-flanked aphVIII expression cassette. Among 46 transformants carrying the CrCRE expression cassette stably, the excision of aphVIII by CrCre recombinase was observed only in one transformant. We then constructed an expression cassette of an in-frame fusion of ble to CrCRE via a short linker peptide. The product of ble (Ble) is a bleomycin-binding protein that confers resistance to bleomycin-related antibiotics such as Zeocin and localizes in the nucleus. Therefore, the ble-(linker)-CrCRE fusion protein is expected to localize in the nucleus. When the ble-(linker)-CrCRE expression cassette was integrated into the genome of strain cc4350 carrying a single copy of the loxP-flanked aphVIII expression cassette, CrCre recombinase-mediated excision of the aphVIII expression cassette was observed at a frequency higher than that in stable transformants of the CrCRE expression cassette. Similarly, from strain cc124 carrying a single loxP-flanked aphVIII expression cassette, the aphVIII expression cassette was successfully excised after

Uptake of uranium from sea water by microalgae

International Nuclear Information System (INIS)

Sakaguchi, Takashi; Horikoshi, Takao; Nakajima, Akira

1978-01-01

The uptake of uranium from aqueous systems especially from sea water by various microalgae was investigated. The freshwater microalgae, Chlorella regularis, Scenedesmus bijuga, Scenedesmus chloreloides, Scenedesmus obliquus, Chlamydomonas angulosa, Chlamydomonas reinhardtii, accumulated relatively large amounts of uranium from the solution containing uranium only. The concentration factors of the above mentioned algae were: Chlorella regularis 3930, Chlamydomonas 2330 - 3400, Scenedesmus 803 - 1920. The uptake of uranium from sea water by Chlorella regularis was inhibited markedly by the co-existence of carbonate ions. Chlorella cells could take up a great quantity of uranium from decarbonated sea water. The uptake of uranium was affected by the pH of sea water, and the amount of uranium absorbed was maximum at pH 5. The experiment was carried out to screen marine microalgae which have the ability to accumulate a large amount of uranium from sea water. The uptake of uranium from sea water by marine microalgae of different species turned out to be in the following decreasing order: Synechococcus > Chlamydomonas >> Chlorella > Dunaliella > Platymonas > Calothrix > Porphyridium. The amount of uranium absorbed differed markedly with different species of marine microalgae. (author)

Transcriptome-Based Identification of the Desiccation Response Genes in Marine Red Algae Pyropia tenera (Rhodophyta) and Enhancement of Abiotic Stress Tolerance by PtDRG2 in Chlamydomonas.

Science.gov (United States)

Im, Sungoh; Lee, Ha-Nul; Jung, Hyun Shin; Yang, Sunghwan; Park, Eun-Jeong; Hwang, Mi Sook; Jeong, Won-Joong; Choi, Dong-Woog

2017-06-01

Pyropia tenera (Kjellman) are marine red algae that grow in the intertidal zone and lose more than 90% of water during hibernal low tides every day. In order to identify the desiccation response gene (DRG) in P. tenera, we generated 1,444,210 transcriptome sequences using the 454-FLX platform from the gametophyte under control and desiccation conditions. De novo assembly of the transcriptome reads generated 13,170 contigs, covering about 12 Mbp. We selected 1160 differentially expressed genes (DEGs) in response to desiccation stress based on reads per kilobase per million reads (RPKM) expression values. As shown in green higher plants, DEGs under desiccation are composed of two groups of genes for gene regulation networks and functional proteins for carbohydrate metabolism, membrane perturbation, compatible solutes, and specific proteins similar to higher plants. DEGs that show no significant homology with known sequences in public databases were selected as DRGs in P. tenera. PtDRG2 encodes a novel polypeptide of 159 amino acid residues locating chloroplast. When PtDRG2 was overexpressed in Chlamydomonas, the PtDRG2 confer mannitol and salt tolerance in transgenic cells. These results suggest that Pyropia may possess novel genes that differ from green plants, although the desiccation tolerance mechanism in red algae is similar to those of higher green plants. These transcriptome sequences will facilitate future studies to understand the common processes and novel mechanisms involved in desiccation stress tolerance in red algae.

Tolerance to cadmium in Chlamydomonas sp. (Chlorophyta) strains isolated from an extreme acidic environment, the Tinto River (SW, Spain)

International Nuclear Information System (INIS)

Aguilera, Angeles; Amils, Ricardo

2005-01-01

The effects of selected concentrations of Cd on the growth and ultrastructure of three strains of Chlamydomonas sp. isolated from a highly acidic river, Rio Tinto (SW Spain) were examined. The river is characterized by its extreme physico-chemical conditions in terms of low pH, mean 2.2 and high concentrations of heavy metals. Growth, Cd accumulation, chlorophyll a, influence of Fe in Cd toxicity and ultrastructural localization were determined. The strains were cultured in both, artificial chemically defined media as well as in natural water from the river. Since iron is the main component of the river water, the effect of different concentrations of this element in relation with Cd toxicity was also analysed. The three strains analysed showed comparable growth and ultrastructural changes. Cd concentration corresponding to 50% growth inhibition (EC 5 ) was 0.2 mM when cells were grown in artificial media. When cells were grown in natural water, no significant differences were found between the controls and the Cd supplemented media even at the highest concentration of 0.8 mM. At an inhibitory level of 0.1 mM of Cd, increasing the concentration of iron up to 90 or 180 mM resulted in a dramatic recovery in algal growth rates in artificial media, reaching normal growth curves. The accumulation of Cd depended on dose and time in the artificial media. The maximal accumulation of Cd was reached after 3 days for all Cd doses, and remained almost unchanged in the subsequent period of time. Chlorophyll a amount depended on dose but not on time in the artificial growth media. At the ultrastructural level, an increase in the periplasmalemmal space was observed due to the presence of a large number of vacuoles, together with a decrease in the relative volume of the nucleus when the cells were incubated in the presence of Cd. Pyrenoid and starch granules were observed and accumulation of spherical electron-dense bodies were also detected. X-ray spectra of these bodies for

Tolerance to cadmium in Chlamydomonas sp. (Chlorophyta) strains isolated from an extreme acidic environment, the Tinto River (SW, Spain)

Energy Technology Data Exchange (ETDEWEB)

Aguilera, Angeles [Centro de Astrobiologia, Instituto Nacional de Tecnica Aeroespacial, Carretera de Ajalvir Km 4, Torrejon de Ardoz, 28850 Madrid (Spain)]. E-mail: aaguilera@cbm.uam.es; Amils, Ricardo [Centro de Astrobiologia, Instituto Nacional de Tecnica Aeroespacial, Carretera de Ajalvir Km 4, Torrejon de Ardoz, 28850 Madrid (Spain); Centro de Biologia Molecular (UAM-CSIC), Universidad Autonoma de Madrid, Canto Blanco, 28049 Madrid (Spain)

2005-11-30

The effects of selected concentrations of Cd on the growth and ultrastructure of three strains of Chlamydomonas sp. isolated from a highly acidic river, Rio Tinto (SW Spain) were examined. The river is characterized by its extreme physico-chemical conditions in terms of low pH, mean 2.2 and high concentrations of heavy metals. Growth, Cd accumulation, chlorophyll a, influence of Fe in Cd toxicity and ultrastructural localization were determined. The strains were cultured in both, artificial chemically defined media as well as in natural water from the river. Since iron is the main component of the river water, the effect of different concentrations of this element in relation with Cd toxicity was also analysed. The three strains analysed showed comparable growth and ultrastructural changes. Cd concentration corresponding to 50% growth inhibition (EC{sub 5}) was 0.2 mM when cells were grown in artificial media. When cells were grown in natural water, no significant differences were found between the controls and the Cd supplemented media even at the highest concentration of 0.8 mM. At an inhibitory level of 0.1 mM of Cd, increasing the concentration of iron up to 90 or 180 mM resulted in a dramatic recovery in algal growth rates in artificial media, reaching normal growth curves. The accumulation of Cd depended on dose and time in the artificial media. The maximal accumulation of Cd was reached after 3 days for all Cd doses, and remained almost unchanged in the subsequent period of time. Chlorophyll a amount depended on dose but not on time in the artificial growth media. At the ultrastructural level, an increase in the periplasmalemmal space was observed due to the presence of a large number of vacuoles, together with a decrease in the relative volume of the nucleus when the cells were incubated in the presence of Cd. Pyrenoid and starch granules were observed and accumulation of spherical electron-dense bodies were also detected. X-ray spectra of these bodies for

A millifluidic study of cell-to-cell heterogeneity in growth-rate and cell-division capability in populations of isogenic cells of Chlamydomonas reinhardtii.

Directory of Open Access Journals (Sweden)

Shima P Damodaran

Full Text Available To address possible cell-to-cell heterogeneity in growth dynamics of isogenic cell populations of Chlamydomonas reinhardtii, we developed a millifluidic drop-based device that not only allows the analysis of populations grown from single cells over periods of a week, but is also able to sort and collect drops of interest, containing viable and healthy cells, which can be used for further experimentation. In this study, we used isogenic algal cells that were first synchronized in mixotrophic growth conditions. We show that these synchronized cells, when placed in droplets and kept in mixotrophic growth conditions, exhibit mostly homogeneous growth statistics, but with two distinct subpopulations: a major population with a short doubling-time (fast-growers and a significant subpopulation of slowly dividing cells (slow-growers. These observations suggest that algal cells from an isogenic population may be present in either of two states, a state of restricted division and a state of active division. When isogenic cells were allowed to propagate for about 1000 generations on solid agar plates, they displayed an increased heterogeneity in their growth dynamics. Although we could still identify the original populations of slow- and fast-growers, drops inoculated with a single progenitor cell now displayed a wider diversity of doubling-times. Moreover, populations dividing with the same growth-rate often reached different cell numbers in stationary phase, suggesting that the progenitor cells differed in the number of cell divisions they could undertake. We discuss possible explanations for these cell-to-cell heterogeneities in growth dynamics, such as mutations, differential aging or stochastic variations in metabolites and macromolecules yielding molecular switches, in the light of single-cell heterogeneities that have been reported among isogenic populations of other eu- and prokaryotes.

Assessing bio-available silver released from silver nanoparticles embedded in silica layers using the green algae Chlamydomonas reinhardtii as bio-sensors

Energy Technology Data Exchange (ETDEWEB)

Pugliara, Alessandro [nMat group-CEMES (Centre d' Elaboration de Matériaux et d' Etudes Structurales)-CNRS, Université de Toulouse, 29 rue Jeanne Marvig, BP 94347, F-31055 Toulouse Cedex 4 (France); LAPLACE (LAboratoire PLAsma et Conversion d' Energie), Université de Toulouse, CNRS, UPS, INPT, 118 route de Narbonne, F-31062 Toulouse (France); Makasheva, Kremena; Despax, Bernard [LAPLACE (LAboratoire PLAsma et Conversion d' Energie), Université de Toulouse, CNRS, UPS, INPT, 118 route de Narbonne, F-31062 Toulouse (France); Bayle, Maxime; Carles, Robert; Benzo, Patrizio; BenAssayag, Gérard; Pécassou, Béatrice [nMat group-CEMES (Centre d' Elaboration de Matériaux et d' Etudes Structurales)-CNRS, Université de Toulouse, 29 rue Jeanne Marvig, BP 94347, F-31055 Toulouse Cedex 4 (France); Sancho, Maria Carmen; Navarro, Enrique [IPE (Instituto Pirenaico de Ecología)-CSIC, Avda. Montañana 1005, Zaragoza 50059 (Spain); Echegoyen, Yolanda [I3A, Department of Analytical Chemistry, University of Zaragoza, C/ María de Luna 3, 50018, Zaragoza (Spain); Bonafos, Caroline, E-mail: bonafos@cemes.fr [nMat group-CEMES (Centre d' Elaboration de Matériaux et d' Etudes Structurales)-CNRS, Université de Toulouse, 29 rue Jeanne Marvig, BP 94347, F-31055 Toulouse Cedex 4 (France)

2016-09-15

Silver nanoparticles (AgNPs) because of their strong antibacterial activity are widely used in health-care sector and industrial applications. Their huge surface-volume ratio enhances the silver release compared to the bulk material, leading to an increased toxicity for microorganisms sensitive to this element. This work presents an assessment of the toxic effect on algal photosynthesis due to small (size < 20 nm) AgNPs embedded in silica layers. Two physical approaches were originally used to elaborate the nanocomposite structures: (i) low energy ion beam synthesis and (ii) combined silver sputtering and plasma polymerization. These techniques allow elaboration of a single layer of AgNPs embedded in silica films at defined nanometer distances (from 0 to 7 nm) beneath the free surface. The structural and optical properties of the nanostructures were studied by transmission electron microscopy and optical reflectance. The silver release from the nanostructures after 20 h of immersion in buffered water was measured by inductively coupled plasma mass spectrometry and ranges between 0.02 and 0.49 μM. The short-term toxicity of Ag to photosynthesis of Chlamydomonas reinhardtii was assessed by fluorometry. The obtained results show that embedding AgNPs reduces the interactions with the buffered water free media, protecting the AgNPs from fast oxidation. The release of bio-available silver (impacting on the algal photosynthesis) is controlled by the depth at which AgNPs are located for a given host matrix. This provides a procedure to tailor the toxicity of nanocomposites containing AgNPs. - Highlights: • Controlled synthesis of 2D arrays of silver nanoparticles embedded in silica. • Assessing bio-available silver release using the green algae as bio-sensors. • The Ag release can be controlled by the distance nanoparticles/dielectric surface. • All the Ag released in solution is in the form of Ag{sup +} ions. • Toxicity comparable to similar concentrations of

Multi-Pixel Photon Counters for Optofluidic Characterization of Particles and Microalgae

Directory of Open Access Journals (Sweden)

Pouya Asrar

2015-06-01

Full Text Available We have developed an optofluidic biosensor to study microscale particles and different species of microalgae. The system is comprised of a microchannel with a set of chevron-shaped grooves. The chevrons allows for hydrodynamic focusing of the core stream in the center using a sheath fluid. The device is equipped with a new generation of highly sensitive photodetectors, multi-pixel photon counter (MPPC, with high gain values and an extremely small footprint. Two different sizes of high intensity fluorescent microspheres and three different species of algae (Chlamydomonas reinhardtii strain 21 gr, Chlamydomonas suppressor, and Chlorella sorokiniana were studied. The forward scattering emissions generated by samples passing through the interrogation region were carried through a multimode fiber, located in 135 degree with respect to the excitation fiber, and detected by a MPPC. The signal outputs obtained from each sample were collected using a data acquisition system and utilized for further statistical analysis. Larger particles or cells demonstrated larger peak height and width, and consequently larger peak area. The average signal output (integral of the peak for Chlamydomonas reinhardtii strain 21 gr, Chlamydomonas suppressor, and Chlorella sorokiniana falls between the values found for the 3.2 and 10.2 μm beads. Different types of algae were also successfully characterized.

pH modulates transport rates of manganese and cadmium in the green alga Chlamydomonas reinhardtii through non-competitive interactions: Implications for an algal BLM

International Nuclear Information System (INIS)

Francois, Laura; Fortin, Claude; Campbell, Peter G.C.

2007-01-01

The influence of pH on short-term uptake of manganese and cadmium by the green alga Chlamydomonas reinhardtii was studied to better understand the nature of proton interactions with metal membrane transporters. Manganese and cadmium internalization fluxes (J int ) were measured over a wide range of free metal ion concentrations from 1 x 10 -10 to 4 x 10 -4 M at several pH values (Mn: 5.0, 6.5 and 8.0; Cd: 5.0 and 6.5). For both metals, first-order biological internalization kinetics were observed but the maximum transport flux (J max ) decreased when pH decreased, in contradiction with the Biotic Ligand Model (BLM). This result suggested a non-competitive inhibition of metal uptake by the H + -ion. A Michaelis-Menten type inhibition model considering proton and calcium competition was tested. The metal biotic ligand stability constants and the stability constants for competitive binding of Ca 2+ and H + with the metal transporters were calculated: for manganese, K Mn = 10 4.20 and K Ca = 10 3.71 ; for cadmium, K Cd = 10 4.19 and K Ca = 10 4.76 ; for both metal transport systems, K H was not a significant parameter. Furthermore, metal uptake was not significantly influenced by the pH of the antecedent growth medium, suggesting that increases in metal fluxes as the pH is raised are caused by conformational changes of the surface transport proteins rather than by the synthesis of additional transport sites. Our results demonstrate that the BLM in its present state does not properly describe the true influence of pH on manganese and cadmium uptake by algae and that a non-competitive inhibition component must be integrated

Acute effects of a prooxidant herbicide on the microalga Chlamydomonas reinhardtii: Screening cytotoxicity and genotoxicity endpoints

International Nuclear Information System (INIS)

Esperanza, Marta; Cid, Ángeles; Herrero, Concepción; Rioboo, Carmen

2015-01-01

Highlights: • Mitochondrial membrane potential constituted the most sensitive parameter assayed. • Several genotoxicity methods were applied for first time in ecotoxicological studies. • Oxidative DNA base damage (8-OHdG) was induced by paraquat exposure. • Cells with DNA strand breakage and subG1-nuclei increased in treated cultures. • Typical apoptosis hallmarks were observed in microalgal cells exposed to paraquat. - Abstract: Since recent evidence has demonstrated that many types of chemicals exhibit oxidative and/or genotoxic potential on living organisms, reactive oxygen species (ROS) formation and DNA damage are currently the best accepted paradigms to assess the potential hazardous biological effects of a wide range of contaminants. The goal of this study was to evaluate the sensitivity of different cytotoxicity and genotoxicity responses on the model microalga Chlamydomonas reinhardtii exposed to the prooxidant herbicide paraquat. In addition to the growth endpoint, cell viability, mitochondrial membrane potential and presence of reactive oxygen species (ROS) were assayed as potential markers of cytotoxicity using flow cytometry (FCM). To study the effects of paraquat on C. reinhardtii DNA, several genotoxicity approaches were implemented for the first time in an ecotoxicological study on microalgae. Oxidative DNA base damage was analysed by measuring the oxidative DNA lesion 8-OHdG by FCM. DNA fragmentation was analysed by different methods: comet assay, and cell cycle analysis by FCM, with a particular focus on the presence of subG1-nuclei. Finally, effects on morphology of nuclei were monitored through DAPI staining. The evaluation of these endpoints showed that several physiological and biochemical parameters reacted to oxidative stress disturbances with greater sensitivity than integrative parameters such as growth rates or cell viability. The experiments revealed concentration-dependent cytotoxicity (ROS formation, depolarization of

Acute effects of a prooxidant herbicide on the microalga Chlamydomonas reinhardtii: Screening cytotoxicity and genotoxicity endpoints

Energy Technology Data Exchange (ETDEWEB)

Esperanza, Marta; Cid, Ángeles; Herrero, Concepción; Rioboo, Carmen, E-mail: carmen.rioboo@udc.es

2015-08-15

Highlights: • Mitochondrial membrane potential constituted the most sensitive parameter assayed. • Several genotoxicity methods were applied for first time in ecotoxicological studies. • Oxidative DNA base damage (8-OHdG) was induced by paraquat exposure. • Cells with DNA strand breakage and subG1-nuclei increased in treated cultures. • Typical apoptosis hallmarks were observed in microalgal cells exposed to paraquat. - Abstract: Since recent evidence has demonstrated that many types of chemicals exhibit oxidative and/or genotoxic potential on living organisms, reactive oxygen species (ROS) formation and DNA damage are currently the best accepted paradigms to assess the potential hazardous biological effects of a wide range of contaminants. The goal of this study was to evaluate the sensitivity of different cytotoxicity and genotoxicity responses on the model microalga Chlamydomonas reinhardtii exposed to the prooxidant herbicide paraquat. In addition to the growth endpoint, cell viability, mitochondrial membrane potential and presence of reactive oxygen species (ROS) were assayed as potential markers of cytotoxicity using flow cytometry (FCM). To study the effects of paraquat on C. reinhardtii DNA, several genotoxicity approaches were implemented for the first time in an ecotoxicological study on microalgae. Oxidative DNA base damage was analysed by measuring the oxidative DNA lesion 8-OHdG by FCM. DNA fragmentation was analysed by different methods: comet assay, and cell cycle analysis by FCM, with a particular focus on the presence of subG1-nuclei. Finally, effects on morphology of nuclei were monitored through DAPI staining. The evaluation of these endpoints showed that several physiological and biochemical parameters reacted to oxidative stress disturbances with greater sensitivity than integrative parameters such as growth rates or cell viability. The experiments revealed concentration-dependent cytotoxicity (ROS formation, depolarization of

Engineering Photosynthetic Organisms for the Production of Renewable Energy Products and Environmental Remediation

Science.gov (United States)

2016-11-25

Standard Form 298 (Rev 8/98) Prescribed by ANSI Std. Z39.18 Final Report W911NF-15-1-0055 66342-CH-REP.1 304-414-4062 a. REPORT 14. ABSTRACT 16...centrifuge, Helios gene gun system, gene pulser, qPCR system, bomb calorimeter system, autoclave, orbit shaker, biosafety cabinet, freeze drier... standard growth conditions. Plants were irrigated with no additional nutrients and grown for 8–10 weeks. WT microalgae Chlamydomonas (Chlamydomonas

Linoleic acid-induced ultra-weak photon emission from Chlamydomonas reinhardtii as a tool for monitoring of lipid peroxidation in the cell membranes.

Directory of Open Access Journals (Sweden)

Ankush Prasad

Full Text Available Reactive oxygen species formed as a response to various abiotic and biotic stresses cause an oxidative damage of cellular component such are lipids, proteins and nucleic acids. Lipid peroxidation is considered as one of the major processes responsible for the oxidative damage of the polyunsaturated fatty acid in the cell membranes. Various methods such as a loss of polyunsaturated fatty acids, amount of the primary and the secondary products are used to monitor the level of lipid peroxidation. To investigate the use of ultra-weak photon emission as a non-invasive tool for monitoring of lipid peroxidation, the involvement of lipid peroxidation in ultra-weak photon emission was studied in the unicellular green alga Chlamydomonas reinhardtii. Lipid peroxidation initiated by addition of exogenous linoleic acid to the cells was monitored by ultra-weak photon emission measured with the employment of highly sensitive charged couple device camera and photomultiplier tube. It was found that the addition of linoleic acid to the cells significantly increased the ultra-weak photon emission that correlates with the accumulation of lipid peroxidation product as measured using thiobarbituric acid assay. Scavenging of hydroxyl radical by mannitol, inhibition of intrinsic lipoxygenase by catechol and removal of molecular oxygen considerably suppressed ultra-weak photon emission measured after the addition of linoleic acid. The photon emission dominated at the red region of the spectrum with emission maximum at 680 nm. These observations reveal that the oxidation of linoleic acid by hydroxyl radical and intrinsic lipoxygenase results in the ultra-weak photon emission. Electronically excited species such as excited triplet carbonyls are the likely candidates for the primary excited species formed during the lipid peroxidation, whereas chlorophylls are the final emitters of photons. We propose here that the ultra-weak photon emission can be used as a non

Protein (Viridiplantae): 159470305 [PGDBj - Ortholog DB

Lifescience Database Archive (English)

Full Text Available predicted protein Chlamydomonas reinhardtii MSSRPKRAASANMANVIAAEKANKAAALHAWPKMWATKLEAQLQLMFMPTRLHRRPLHQGTCRNYSTAPGITGVIELTSAFYRMYPNATFVFNKETAAKGTYRGEEETAASWWLKHVGSKLEIYLSPLRCRPEVSR ...

Coleção de microalgas de ambientes dulciaquícolas naturais da Bahia, Brasil, como potencial fonte para a produção de biocombustíveis: uma abordagem taxonômica Collection of microalgae from natural freshwater environments of Bahia, Brazil, as a potential source for biofuel production: a taxonomic approach

Directory of Open Access Journals (Sweden)

Maria Cristina de Queiroz Mendes

2012-09-01

Full Text Available O presente trabalho envolveu a identificação taxonômica de espécies nativas de microalgas (isoladas de ecossistemas dulciaquícolas localizados nos arredores de Salvador, Bahia integrantes da Coleção de Microalgas dulciaquícolas do LABIOMAR/IB/UFBA, visando estudos taxonômicos mais aprofundados (ultraestruturais e moleculares e experimentos que possam avaliar sua capacidade para suprir cadeias produtivas de biocombustíveis. As coletas foram realizadas nos arredores de Salvador, Bahia, Brasil. A identificação das espécies foi efetuada com base em caracteres morfológicos. Foram identificados 19 táxons, 12 em nível de espécie e nove em nível de gênero, sendo 14 Chlorophyceae (Chlamydomonas sp1, Chlamydomonas sp2, Chlamydomonas sp3, Chlamydocapsa bacillus (Teiling Fott, Chlorococcum sp1, Chlorococcum sp2, Coelastrum indicum Turn.. Coelastrum microporum Nägeli, Desmodesmus brasiliensis (Bohl. Hegew, Scenedesmum obliquus (Turpin Kütz, Ankistrodesmus falcatus (Corda Ralfs, Ankistrodesmus fusiformis Corda, Kirchneriella lunaris (Kirchner. Möbius, Pseudokirchneriella subcapitata (Korshikov F. Hindák, três Trebouxiophyceae (Botryococcus braunii Kütz., Botryococcus terribilis Komárek et Marvan e Chlorella vulgaris Beijerinck, uma Bacillariophyceae (Nitzschia sp. e uma Cyanobacteria (Synechocystis sp..This study identified native species of microalgae (maintained at LABIOMAR/IB/UFBA Collection of Freshwater Microalgae to indicate their potential to supply the biofuel production chain. Samples were collected in freshwater ecosystems around Salvador, Bahia, Brazil. Species identification was based in morphological characteristics. Nineteen species were isolated and identified, 12 at the level of species and nine at the level of genus: 14 Chlorophyceae (Chlamydomonas sp1, Chlamydomonas sp2, Chlamydomonas sp3, Chlamydocapsa bacillus (Teiling Fott, Chlorococcum sp1, Chlorococcum sp2, Coelastrum indicum Turn. Coelastrum microporum N�

Evaluation of three herbicide resistance genes for use in genetic transformations and for potential crop protection in algae production.

Science.gov (United States)

Brueggeman, Andrew J; Kuehler, Daniel; Weeks, Donald P

2014-09-01

Genes conferring resistance to the herbicides glyphosate, oxyfluorfen and norflurazon were developed and tested for use as dominant selectable markers in genetic transformation of Chlamydomonas reinhardtii and as potential tools for the protection of commercial-scale algal production facilities against contamination by organisms sensitive to these broad-spectrum herbicides. A synthetic glyphosate acetyltransferase (GAT) gene, when fitted with a strong Chlamydomonas promoter, conferred a 2.7×-fold increase in tolerance to the EPSPS inhibitor, glyphosate, in transgenic cells compared with progenitor WT cells. A mutant Chlamydomonas protoporphyrinogen oxidase (protox, PPO) gene previously shown to produce an enzyme insensitive to PPO-inhibiting herbicides, when genetically engineered, generated transgenic cells able to tolerate up to 136× higher levels of the PPO inhibitor, oxyfluorfen, than nontransformed cells. Genetic modification of the Chlamydomonas phytoene desaturase (PDS) gene-based gene sequences found in various norflurazon-resistant organisms allowed production of transgenic cells tolerant to 40× higher levels of norflurazon than nontransgenic cells. The high efficiency of all three herbicide resistance genes in producing transgenic cells demonstrated their suitability as dominant selectable markers for genetic transformation of Chlamydomonas and, potentially, other eukaryotic algae. However, the requirement for high concentrations of glyphosate and its associated negative effects on cell growth rates preclude its consideration for use in large-scale production facilities. In contrast, only low doses of norflurazon and oxyfluorfen (~1.5 μm and ~0.1 μm, respectively) are required for inhibition of cell growth, suggesting that these two herbicides may prove effective in large-scale algal production facilities in suppressing growth of organisms sensitive to these herbicides. © 2014 Society for Experimental Biology, Association of Applied Biologists and

EST analysis of the scaly green flagellate Mesostigma viride (Streptophyta: Implications for the evolution of green plants (Viridiplantae

Directory of Open Access Journals (Sweden)

Melkonian Michael

2006-02-01

Full Text Available Abstract Background The Viridiplantae (land plants and green algae consist of two monophyletic lineages, the Chlorophyta and the Streptophyta. The Streptophyta include all embryophytes and a small but diverse group of freshwater algae traditionally known as the Charophyceae (e.g. Charales, Coleochaete and the Zygnematales. The only flagellate currently included in the Streptophyta is Mesostigma viride Lauterborn. To gain insight into the genome evolution in streptophytes, we have sequenced 10,395 ESTs from Mesostigma representing 3,300 independent contigs and compared the ESTs of Mesostigma with available plant genomes (Arabidopsis, Oryza, Chlamydomonas, with ESTs from the bryophyte Physcomitrella, the genome of the rhodophyte Cyanidioschyzon, the ESTs from the rhodophyte Porphyra, and the genome of the diatom Thalassiosira. Results The number of expressed genes shared by Mesostigma with the embryophytes (90.3 % of the expressed genes showing similarity to known proteins is higher than with Chlamydomonas (76.1 %. In general, cytosolic metabolic pathways, and proteins involved in vesicular transport, transcription, regulation, DNA-structure and replication, cell cycle control, and RNA-metabolism are more conserved between Mesostigma and the embryophytes than between Mesostigma and Chlamydomonas. However, plastidic and mitochondrial metabolic pathways, cytoskeletal proteins and proteins involved in protein folding are more conserved between Mesostigma and Chlamydomonas than between Mesostigma and the embryophytes. Conclusion Our EST-analysis of Mesostigma supports the notion that this organism should be a suitable unicellular model for the last flagellate common ancestor of the streptophytes. Mesostigma shares more genes with the embryophytes than with the chlorophyte Chlamydomonas reinhardtii, although both organisms are flagellate unicells. Thus, it seems likely that several major physiological changes (e.g. in the regulation of photosynthesis

New features on the environmental regulation of metabolism revealed by modeling the cellular proteomic adaptations induced by light, carbon and inorganic nitrogen in Chlamydomonas reinhardtii

Directory of Open Access Journals (Sweden)

Stéphanie Gérin

2016-08-01

Full Text Available Microalgae are currently emerging to be very promising organisms for the production of biofuels and high-added value compounds. Understanding the influence of environmental alterations on their metabolism is a crucial issue. Light, carbon and nitrogen availability have been reported to induce important metabolic adaptations. So far, the influence of these variables has essentially been studied while varying only one or two environmental factors at the same time. The goal of the present work was to model the cellular proteomic adaptations of the green microalga Chlamydomonas reinhardtii upon the simultaneous changes of light intensity, carbon concentrations (CO2 and acetate and inorganic nitrogen concentrations (nitrate and ammonium in the culture medium. Statistical design of experiments (DOE enabled to define 32 culture conditions to be tested experimentally. Relative protein abundance was quantified by two dimensional differential in-gel electrophoresis (2D-DIGE. Additional assays for respiration, photosynthesis, and lipid and pigment concentrations were also carried out. A hierarchical clustering survey enabled to partition biological variables (proteins + assays into eight co-regulated clusters. In most cases, the biological variables partitioned in the same cluster had already been reported to participate to common biological functions (acetate assimilation, bioenergetic processes, light harvesting, Calvin cycle and protein metabolism. The environmental regulation within each cluster was further characterized by a series of multivariate methods including principal component analysis and multiple linear regressions. This metadata analysis enabled to highlight the existence of a clear regulatory pattern for every cluster and to mathematically simulate the effects of light, carbon and nitrogen. The influence of these environmental variables on cellular metabolism is described in details and thoroughly discussed. This work provides an overview

Effect of chromium oxide (III) nanoparticles on the production of reactive oxygen species and photosystem II activity in the green alga Chlamydomonas reinhardtii

International Nuclear Information System (INIS)

Costa, Cristina Henning da; Perreault, François; Oukarroum, Abdallah; Melegari, Sílvia Pedroso; Popovic, Radovan; Matias, William Gerson

2016-01-01

With the growth of nanotechnology and widespread use of nanomaterials, there is an increasing risk of environmental contamination by nanomaterials. However, the potential implications of such environmental contamination are hard to evaluate since the toxicity of nanomaterials if often not well characterized. The objective of this study was to evaluate the toxicity of a chromium-based nanoparticle, Cr_2O_3-NP, used in a wide diversity of industrial processes and commercial products, on the unicellular green alga Chlamydomonas reinhardtii. The deleterious impacts of Cr_2O_3-NP were characterized using cell density measurements, production of reactive oxygen species (ROS), esterase enzymes activity, and photosystem II electron transport as indicators of toxicity. Cr_2O_3-NP exposure inhibited culture growth and significantly lowered cellular Chlorophyll a content. From cell density measurements, EC50 values of 2.05 ± 0.20 and 1.35 ± 0.06 g L"−"1 Cr_2O_3-NP were obtained after 24 and 72 h of exposure, respectively. In addition, ROS levels were increased to 160.24 ± 2.47% and 59.91 ± 0.15% of the control value after 24 and 72 h of exposition to 10 g L"−"1 Cr_2O_3-NP. At 24 h of exposure, the esterase activity increased to 160.24% of control value, revealing a modification of the short-term metabolic response of algae to Cr_2O_3-NP exposure. In conclusion, the metabolism of C. reinhardtii was the most sensitive to Cr_2O_3-NP after 24 h of treatment. - Highlights: • Cr_2O_3 nanoparticles are unstable and form large aggregates in the medium. • EC50 for growth inhibition of C. reinhardtii is 1.35 g L"−"1 at 72 h. • Cr_2O_3 nanoparticles increase ROS levels at 10 g L"−"1. • Cr_2O_3 nanoparticles affect photosynthetic electron transport.

Protein (Viridiplantae): 159468384 [PGDBj - Ortholog DB

Lifescience Database Archive (English)

Full Text Available 3436 hypothetical protein CHLREDRAFT_180911 Chlamydomonas reinhardtii MTTEEPLSCSKIRSWNITVYSFTLKGLPGCLEPSHSFWVKEREGEWGLKCLSETFSHELVENVPGREEVSNLLKKGGSSNKSQKGGWICCERNCFLCQHKKCQVLI ...

Photobiological hydrogen production with the unicellular green alga Chlamydomonas reinhardtii under process engineering aspects; Photobiologische Wasserstoffproduktion mit der einzelligen Gruenalge Chlamydomonas reinhardtii unter verfahrenstechnischen Aspekten

Energy Technology Data Exchange (ETDEWEB)

Geier, Stephanie

2011-07-01

Hydrogen is of high interest as a clean and environmentally friendly energy source as its combustion only emits water and energy. However, currently hydrogen is produced in energy demanding processes by the consumption of fossil fuels. An alternative way of sustainable and non-polluting hydrogen production could be provided by use of photosynthetic active microalgae. Within this work, the photobiological hydrogen production with the unicellular green algae Chlamydomonas reinhardtii is investigated under the aspects of bioprocess-engineering and economics. Objectives are, besides the increase of the photochemical efficiency, the cultivation of the algae and subsequent hydrogen production under cost-free sunlight. It could be demonstrated that outdoor cultivation of C. reinhardtii is possible in Central Europe throughout the year by using e.g. waste heat. Similar cell numbers in the range from 1,2.10{sup 7} cells ml{sup -1} to 1,7.10{sup 7} cells ml{sup -1} could be achieved in closed photobioreactors of the type Photobioreactor Screening Module under controlled laboratory conditions and both continuous illumination (200 {mu}mol.m{sup -2}.s{sup -1}) and simulated outdoor conditions according to the light intensity of idealized summer day as well as in outdoor experiments (up to 2000 {mu}mol.m{sup -2}.s{sup -1}).The use of 10 % CO{sub 2} corresponding to the CO{sub 2} content in flue gas led to a doubling of cell numbers under continuous illumination to 4,2.10{sup 7} cells ml{sup -1}, compared to the reference culture bubbled with 3 % CO{sub 2}. A significant increase of cell numbers under the light profiles of an idealized summer day could not be achieved. The cultivation under the light profile of a winter day at 25 C reduced cell growth to 54 %, compared to the summer simulation. In open 30 L outdoor ponds, only 0,26.10{sup 7} cells ml{sup -1} could be achieved under photoheterotrophic conditions during the summer months, which corresponds to 20 % of the cell

Protein (Viridiplantae): 159466610 [PGDBj - Ortholog DB

Lifescience Database Archive (English)

Full Text Available 2419 hypothetical protein CHLREDRAFT_123820, partial Chlamydomonas reinhardtii RVQCRLVDMPAPCLPPFLPTCPHKPRRIPMPCTDAH...ELVDMPAPCLPPFLPDNLPARAPQAPHAVTDAHECMQCRLVDMPAPCLPPFLPKCPHKPRRLPMPCTDAHECNMPAPCLPPFLPKCPHKPRRLPMPCTDAHECMQCRLVDMPAPCLPAFLPNCPHKPRRLPMPCTDAHECSAGW ...

Transcriptional and cellular effects of benzotriazole UV stabilizers UV-234 and UV-328 in the freshwater invertebrates Chlamydomonas reinhardtii and Daphnia magna.

Science.gov (United States)

Giraudo, Maeva; Cottin, Guillaume; Esperanza, Marta; Gagnon, Pierre; Silva, Amila O De; Houde, Magali

2017-12-01

Benzotriazole ultra violet stabilizers (BZT-UVs) are compounds used in many applications and products to prevent photochemical degradation. Despite their widespread presence in aquatic ecosystems and persistence in the environment, there are very limited data on their effects and toxicity, and their modes of action remain largely unknown. The objectives of the present study were to evaluate the chronic effects of 2 BZT-UVs, 2-(2H-benzotriazol-2-yl)-4,6-bis(1-methyl-1-phenylethyl)phenol (UV-234) and 2-(2H-benzotriazol-2-yl)-4,6-di-tert-pentylphenol (UV-328), on the freshwater green algae Chlamydomonas reinhardtii and the freshwater crustacean Daphnia magna. Organisms were exposed to 0.01 and 10 μg/L of UV-234, UV-328, as well as a mixture of the 2 compounds. Life-history endpoints (viability, reproduction, and growth) and oxidative stress-related biomarkers (gene transcription, reactive oxygen species [ROS] production, and lipid peroxidation) were measured. Daphnia magna growth, reproduction, and gene transcription were not impacted by 21-d individual or mixed exposure. After 96-h of exposure, no differences were observed on the cellular viability of C. reinhardtii for either of the 2 BZT-UVs. In the algae, results showed increased ROS production in response to UV-328 and lipid peroxidation following exposure to UV-234. Synergistic effects of the 2 BZT-UVs were evident at the transcriptional level with 2 to 6 times up-regulation of glutathione peroxidase (gp x ) in response to the mixture for all treatment conditions. The transcription of superoxide dismutase (sod), catalase (cat), and ascorbic peroxidase (apx) was also regulated by UV-234 and UV-328 in the green algae, most likely as a result of ROS production and lipid peroxidation. Results from the present study suggest potential impacts of UV-234 and UV-328 exposure on the antioxidant defense system in C. reinhardtii. Environ Toxicol Chem 2017;36:3333-3342. © 2017 Crown in the Right of Canada. Published by

Knock-Down of the IFR1 Protein Perturbs the Homeostasis of Reactive Electrophile Species and Boosts Photosynthetic Hydrogen Production in Chlamydomonas reinhardtii.

Science.gov (United States)

Venkanna, Deepak; Südfeld, Christian; Baier, Thomas; Homburg, Sarah V; Patel, Anant V; Wobbe, Lutz; Kruse, Olaf

2017-01-01

The protein superfamily of short-chain dehydrogenases/reductases (SDR), including members of the atypical type (aSDR), covers a huge range of catalyzed reactions and in vivo substrates. This superfamily also comprises isoflavone reductase-like (IRL) proteins, which are aSDRs highly homologous to isoflavone reductases from leguminous plants. The molecular function of IRLs in non-leguminous plants and green microalgae has not been identified as yet, but several lines of evidence point at their implication in reactive oxygen species homeostasis. The Chlamydomonas reinhardtii IRL protein IFR1 was identified in a previous study, analyzing the transcriptomic changes occurring during the acclimation to sulfur deprivation and anaerobiosis, a condition that triggers photobiological hydrogen production in this microalgae. Accumulation of the cytosolic IFR1 protein is induced by sulfur limitation as well as by the exposure of C. reinhardtii cells to reactive electrophile species (RES) such as reactive carbonyls. The latter has not been described for IRL proteins before. Over-accumulation of IFR1 in the singlet oxygen response 1 ( sor1 ) mutant together with the presence of an electrophile response element, known to be required for SOR1-dependent gene activation as a response to RES, in the promoter of IFR1 , indicate that IFR1 expression is controlled by the SOR1-dependent pathway. An implication of IFR1 into RES homeostasis, is further implied by a knock-down of IFR1 , which results in a diminished tolerance toward RES. Intriguingly, IFR1 knock-down has a positive effect on photosystem II (PSII) stability under sulfur-deprived conditions used to trigger photobiological hydrogen production, by reducing PSII-dependent oxygen evolution, in C. reinhardtii . Reduced PSII photoinhibition in IFR1 knock-down strains prolongs the hydrogen production phase resulting in an almost doubled final hydrogen yield compared to the parental strain. Finally, IFR1 knock-down could be

Real-time monitoring of genetically modified Chlamydomonas reinhardtii during the Foton M3 space mission and ground irradiation experiment

Science.gov (United States)

Lambreva, Maya; Rea, Giuseppina; Antonacci, Amina; Serafini, Agnese; Damasso, Mario; Margonelli, Andrea; Johanningmeier, Udo; Bertalan, Ivo; Pezzotti, Gianni; Giardi, Maria Teresa

Long-term space exploration, colonization or habitation requires biological life support systems capable to cope with the deleterious space environment. The use of oxygenic photosynthetic microrganisms is an intriguing possibility mainly for food, O2 and nutraceutical compounds production. The critical points of utilizing plantsor algae-based life support systems are the microgravity and the ionizing radiation, which can influence the performance of these organisms. The aim of the present study was to assess the effects of space environment on the photosynthetic activity of various microrganisms and to select space stress-tolerant strains. Site-directed and random mutants of the unicellular green alga Chlamydomonas reinhardtii of Photosystem II D1 protein were used as a model system to test and select the amino acid substitutions capable to account for space stress tolerance. We focussed our studies also on the accumulation of the Photosystem II photoprotective carotenoids (the xantophylls violaxanthin, anteraxanthin and zeaxanthin), powerful antioxidants that epidemiological studies demonstrated to be human vision protectors. Metabolite profiling by quantitative HPLC methods revealed the organisms and the stress conditions capable to accumulate the highest pigment levels. In order to develop a project for a rationale metabolic engineering of algal secondary metabolites overproduction, we are performing expression analyses on the carotenoid biosynthetic pathway under physiological and mimicked space conditions. To identify the consequences of the space environment on the photosynthetic apparatus the changes in the Photosystem II efficiency were monitored in real time during the ESA-Russian Foton-M3 mission in September 2007. For the space flight a high-tech, multicell fluorescence biosensor, Photo-II, was designed and built by the Centre for Advanced Research in Space Optics in collaboration with Kayser-Italy, Biosensor and DAS. Photo-II is an automatic device

SOIL ALGOLOGICAL SINUSIAE OF URBANIZED ECOSYSTEMS OF DONETSK PRIAZOVYE (THE CASE OF MARIUPOL

Directory of Open Access Journals (Sweden)

Shekhovtseva O.G.

2012-12-01

Full Text Available During research of urboekosystem of Donetsk Priazovye 105 types and intraspecific algosoil taxons are revealed: Chlorophyta – 42, Cyanoprocaryota – 34, Xanthophyta – 19,Bacillariophyta – 8, Eustigmatophyta – 2. It is often met: Phormidium autumnale, Chlamydomonas elliptica, Chlamydomonas gloeogama, Palmellopsis gelatinosa, Chlorella minutissima, Сhlorella vulgaris, Stichococcus minor, Stichococcus bacillaris, Bracteacoccus minor, Botrydiopsis arhiza, Pleurochloris magna, Navicula mutica, Navicula pelliculosa, Hantzschia amphioxys. Ecological structure of algogroupping: Ch31 P27 X16 C11 H7 B7 СF3 M2 amph1 (105.

Protein (Viridiplantae): 714399 [PGDBj - Ortholog DB

Lifescience Database Archive (English)

Full Text Available 3051:329 ... 3052:329 ... 3055:329 ... predicted protein Chlamydomonas reinhardtii MAPAALPGRSVKSKQAHLLRTDAHRVKSKQAHLLRTDAHRVKSKQAHLLRTDA...HRVKSKQAHLLRTDAHRVKSKQAHLLRTDAHRVALTTLTGALSLFGGACTATSFVLQVSASAASYAASLRLSCPAVPSLTDVA

Gene : CBRC-MEUG-01-1826 [SEVENS

Lifescience Database Archive (English)

Full Text Available utinin [Chlamydomonas incerta] 0.0 39% MPLCMSTCLTNYISTLCTSCTSVWPPTPAPPALLHTAVCPPTPAPLHTAVCPPTPASPAPLHTAVCPPTPAPLHTAVCPPTPASPAPLHTAVC...PPTPAPLHTAVCPPTPAPPAPLRTAVCPPTPAPLHTAICPPTLAPPAPLHTAFCPPTPAPLHTAFCPPTLAPPAPLHTAVCPPTPAPLHTAFCPPTPAPLHTAVC...PPTPAPLHTAICPPTPAPLRTAICPPTPAPLHTAVCPPTLAPPAPLRTAVCPPTPAPPAPLRTAVCPPTPAPLRTAVC...PPTPAPLRTAICPPTPAPLHTAVCPPTLAPPAPLRTAVCPPTPAPPAPLRTAVCPPTPAPLRTAICPPTPAPLHTAVCPPTLAPPAPLRTAVC...PPTPAPPAPLRTAICPPTPAPLRTAICPPTPAPLHTAVCPPTLAPPAPLRTAVCPPTPAPPAPLRTAVCPPTPAPPAPLCTAVCPPTPAPLCTAVCPPTPAPPAPLCTAVCPPTPAPLCTAVC

Light-induced conformational changes of LOV1 (light oxygen voltage-sensing domain 1) and LOV2 relative to the kinase domain and regulation of kinase activity in Chlamydomonas phototropin.

Science.gov (United States)

Okajima, Koji; Aihara, Yusuke; Takayama, Yuki; Nakajima, Mihoko; Kashojiya, Sachiko; Hikima, Takaaki; Oroguchi, Tomotaka; Kobayashi, Amane; Sekiguchi, Yuki; Yamamoto, Masaki; Suzuki, Tomomi; Nagatani, Akira; Nakasako, Masayoshi; Tokutomi, Satoru

2014-01-03

Phototropin (phot), a blue light (BL) receptor in plants, has two photoreceptive domains named LOV1 and LOV2 as well as a Ser/Thr kinase domain (KD) and acts as a BL-regulated protein kinase. A LOV domain harbors a flavin mononucleotide that undergoes a cyclic photoreaction upon BL excitation via a signaling state in which the inhibition of the kinase activity by LOV2 is negated. To understand the molecular mechanism underlying the BL-dependent activation of the kinase, the photochemistry, kinase activity, and molecular structure were studied with the phot of Chlamydomonas reinhardtii. Full-length and LOV2-KD samples of C. reinhardtii phot showed cyclic photoreaction characteristics with the activation of LOV- and BL-dependent kinase. Truncation of LOV1 decreased the photosensitivity of the kinase activation, which was well explained by the fact that the signaling state lasted for a shorter period of time compared with that of the phot. Small angle x-ray scattering revealed monomeric forms of the proteins in solution and detected BL-dependent conformational changes, suggesting an extension of the global molecular shapes of both samples. Constructed molecular model of full-length phot based on the small angle x-ray scattering data proved the arrangement of LOV1, LOV2, and KD for the first time that showed a tandem arrangement both in the dark and under BL irradiation. The models suggest that LOV1 alters its position relative to LOV2-KD under BL irradiation. This finding demonstrates that LOV1 may interact with LOV2 and modify the photosensitivity of the kinase activation through alteration of the duration of the signaling state in LOV2.

Protein (Viridiplantae): 569482 [PGDBj - Ortholog DB

Lifescience Database Archive (English)

Full Text Available 3051:1120 ... 3052:1120 ... 3055:1120 ... SR protein factor Chlamydomonas reinhardtii MSYRDRDRDRGDRGYSDRDRDRGRDDRRGGDRGGDRGGGGGGDRG...PRDMMRIESKTKGDERRDDRRRSRSRSPRRSSRRSSRSPRRSRSRSPRRSRSPRADRGRDRSPRDRSPRDRSPRDRSPRDRSPRERSPVRVERERSPERERSPERERVREDSRSPPPRERSPPPRDRSPPPRERSPSPRRDSPPRDDYAGDDF

Gene : CBRC-PHAM-01-1650 [SEVENS

Lifescience Database Archive (English)

Full Text Available tinin [Chlamydomonas reinhardtii] 1e-68 36% MFFFPTLSPPPSSPLTLIPSPSQSLLPSPSVPTPSSLHPHLHPSPLTPSSSRLSPPHLICPHPIFIPSILTPSSSHLSPAHPHP...MCPHSHHPHPHPSPLTPSSPHPSPAHPHPMCPHSPHPHPHPSPLTPPSPHPSPAHPHPMCPHSPHPHPMCPHSPHPHPHLSPLTPSSP...PSIPTPSSPPSVLTHPILTPIHPHSPHPHPHPSPLTPSSPHPSPLTPSSPPSVPTHPILTPSVPTHPILTPSVPTPSSPHV...SPLTPSSSPSVPTHPTLTPIHPHSILTPICPHSPHPHPHPSPLTPSSSPSVSTHPILTPIHPHSIFTPICPHSPHPHPHPSPLTPSSPPSVPTHPILTPSIPTHPILTPIRPHSPHPHPIRPHSPHPHP...IRPHPILTPCVPTHPILIPICLHSPHPHPHPSPLHLHPHLSSLTPSSPPSIPTHPILPSSSPPHPCHSSWEAGCTCVEPEPPHPCPSLPSPLAEREGTAWDWLPPVAMTVARIRAVSSPCRKHVMNYGCPIFSERPDL ...

Effect of chromium oxide (III) nanoparticles on the production of reactive oxygen species and photosystem II activity in the green alga Chlamydomonas reinhardtii

Energy Technology Data Exchange (ETDEWEB)

Costa, Cristina Henning da [Department of Sanitary and Environmental Engineering, Federal University of Santa Catarina, Campus Universitário, CEP: 88040-970, Florianópolis, SC (Brazil); Perreault, François [School of Sustainable Engineering and the Built Environment, Arizona State University, Tempe, AZ 85287-3005 (United States); Oukarroum, Abdallah [Department of Chemistry, University of Quebec in Montréal, 2101, Jeanne Mance Street, Station Centre-Ville, Montréal, QC H2X 2J6 (Canada); Melegari, Sílvia Pedroso [Department of Sanitary and Environmental Engineering, Federal University of Santa Catarina, Campus Universitário, CEP: 88040-970, Florianópolis, SC (Brazil); Center of Marine Studies, Federal University of Parana, Beira-mar Avenue, 83255-976, Pontal do Parana, PR (Brazil); Popovic, Radovan [Department of Chemistry, University of Quebec in Montréal, 2101, Jeanne Mance Street, Station Centre-Ville, Montréal, QC H2X 2J6 (Canada); Matias, William Gerson, E-mail: william.g.matias@ufsc.br [Department of Sanitary and Environmental Engineering, Federal University of Santa Catarina, Campus Universitário, CEP: 88040-970, Florianópolis, SC (Brazil)

2016-09-15

With the growth of nanotechnology and widespread use of nanomaterials, there is an increasing risk of environmental contamination by nanomaterials. However, the potential implications of such environmental contamination are hard to evaluate since the toxicity of nanomaterials if often not well characterized. The objective of this study was to evaluate the toxicity of a chromium-based nanoparticle, Cr{sub 2}O{sub 3}-NP, used in a wide diversity of industrial processes and commercial products, on the unicellular green alga Chlamydomonas reinhardtii. The deleterious impacts of Cr{sub 2}O{sub 3}-NP were characterized using cell density measurements, production of reactive oxygen species (ROS), esterase enzymes activity, and photosystem II electron transport as indicators of toxicity. Cr{sub 2}O{sub 3}-NP exposure inhibited culture growth and significantly lowered cellular Chlorophyll a content. From cell density measurements, EC50 values of 2.05 ± 0.20 and 1.35 ± 0.06 g L{sup −1} Cr{sub 2}O{sub 3}-NP were obtained after 24 and 72 h of exposure, respectively. In addition, ROS levels were increased to 160.24 ± 2.47% and 59.91 ± 0.15% of the control value after 24 and 72 h of exposition to 10 g L{sup −1} Cr{sub 2}O{sub 3}-NP. At 24 h of exposure, the esterase activity increased to 160.24% of control value, revealing a modification of the short-term metabolic response of algae to Cr{sub 2}O{sub 3}-NP exposure. In conclusion, the metabolism of C. reinhardtii was the most sensitive to Cr{sub 2}O{sub 3}-NP after 24 h of treatment. - Highlights: • Cr{sub 2}O{sub 3} nanoparticles are unstable and form large aggregates in the medium. • EC50 for growth inhibition of C. reinhardtii is 1.35 g L{sup −1} at 72 h. • Cr{sub 2}O{sub 3} nanoparticles increase ROS levels at 10 g L{sup −1}. • Cr{sub 2}O{sub 3} nanoparticles affect photosynthetic electron transport.

Cyclobutane pyrimidine dimers photolyase from extremophilic microalga: Remarkable UVB resistance and efficient DNA damage repair

Energy Technology Data Exchange (ETDEWEB)

Li, Chongjie [Key Laboratory of Marine Bioactive Substance, The First Institute of Oceanography, State Oceanic Administration, Qingdao 266061 (China); Ma, Li [Key Laboratory of Biofuels, and Shandong Provincial Key Laboratory of Energy Genetics, Qingdao Institute of Bioenergy and Bioprocess Technology, Chinese Academy of Sciences, Qingdao 266101 (China); Mou, Shanli [Yellow Sea Fisheries Research Institute, Chinese Academy of Fishery Sciences, Qingdao (China); Wang, Yibin, E-mail: wangyibin@fio.org.cn [Key Laboratory of Marine Bioactive Substance, The First Institute of Oceanography, State Oceanic Administration, Qingdao 266061 (China); Zheng, Zhou; Liu, Fangming; Qi, Xiaoqing; An, Meiling; Chen, Hao [Key Laboratory of Marine Bioactive Substance, The First Institute of Oceanography, State Oceanic Administration, Qingdao 266061 (China); Miao, Jinlai, E-mail: miaojinlai@163.com [Key Laboratory of Marine Bioactive Substance, The First Institute of Oceanography, State Oceanic Administration, Qingdao 266061 (China); State Key Laboratory of Biological Fermentation Engineering of Beer (In Preparation), Qingdao (China)

2015-03-15

Highlights: • Chlamydomonas sp. ICE-L photolyase gene PHR2 is first cloned and expressed in E. coli. • PHR2 complemented E. coli could efficiently survival from UV radiation. • Expressed PHR2 photolyase has distinct photo-reactivation activity in vitro. - Abstract: Bacteria living in the Antarctic region have developed several adaptive features for growth and survival under extreme conditions. Chlamydomonas sp. ICE-Lis well adapted to high levels of solar UV radiation. A putative photolyase was identified in the Chlamydomonas sp. ICE-L transcriptome. The complete cDNA sequence was obtained by RACE-PCR. This PHR encoding includes a polypeptide of 579 amino acids with clear photolyase signatures belonging to class II CPD-photolyases, sharing a high degree of homology with Chlamydomonas reinhardtii (68%). Real-time PCR was performed to investigate the potential DNA damage and responses following UVB exposure. CPD photolyase mRNA expression level increased over 50-fold in response to UVB radiation for 6 h. Using photolyase complementation assay, we demonstrated that DNA photolyase increased photo-repair more than 116-fold in Escherichia coli strain SY2 under 100 μw/cm{sup 2} UVB radiation. To determine whether photolyase is active in vitro, CPD photolyase was over-expressed. It was shown that pyrimidine dimers were split by the action of PHR2. This study reports the unique structure and high activity of the enzyme. These findings are relevant for further understanding of molecular mechanisms of photo-reactivation, and will accelerate the utilization of photolyase in the medical field.

Cyclobutane pyrimidine dimers photolyase from extremophilic microalga: Remarkable UVB resistance and efficient DNA damage repair

International Nuclear Information System (INIS)

Li, Chongjie; Ma, Li; Mou, Shanli; Wang, Yibin; Zheng, Zhou; Liu, Fangming; Qi, Xiaoqing; An, Meiling; Chen, Hao; Miao, Jinlai

2015-01-01

Highlights: • Chlamydomonas sp. ICE-L photolyase gene PHR2 is first cloned and expressed in E. coli. • PHR2 complemented E. coli could efficiently survival from UV radiation. • Expressed PHR2 photolyase has distinct photo-reactivation activity in vitro. - Abstract: Bacteria living in the Antarctic region have developed several adaptive features for growth and survival under extreme conditions. Chlamydomonas sp. ICE-Lis well adapted to high levels of solar UV radiation. A putative photolyase was identified in the Chlamydomonas sp. ICE-L transcriptome. The complete cDNA sequence was obtained by RACE-PCR. This PHR encoding includes a polypeptide of 579 amino acids with clear photolyase signatures belonging to class II CPD-photolyases, sharing a high degree of homology with Chlamydomonas reinhardtii (68%). Real-time PCR was performed to investigate the potential DNA damage and responses following UVB exposure. CPD photolyase mRNA expression level increased over 50-fold in response to UVB radiation for 6 h. Using photolyase complementation assay, we demonstrated that DNA photolyase increased photo-repair more than 116-fold in Escherichia coli strain SY2 under 100 μw/cm 2 UVB radiation. To determine whether photolyase is active in vitro, CPD photolyase was over-expressed. It was shown that pyrimidine dimers were split by the action of PHR2. This study reports the unique structure and high activity of the enzyme. These findings are relevant for further understanding of molecular mechanisms of photo-reactivation, and will accelerate the utilization of photolyase in the medical field

Is chloroplastic class IIA aldolase a marine enzyme?

Science.gov (United States)

Miyasaka, Hitoshi; Ogata, Takeru; Tanaka, Satoshi; Ohama, Takeshi; Kano, Sanae; Kazuhiro, Fujiwara; Hayashi, Shuhei; Yamamoto, Shinjiro; Takahashi, Hiro; Matsuura, Hideyuki; Hirata, Kazumasa

2016-01-01

Expressed sequence tag analyses revealed that two marine Chlorophyceae green algae, Chlamydomonas sp. W80 and Chlamydomonas sp. HS5, contain genes coding for chloroplastic class IIA aldolase (fructose-1, 6-bisphosphate aldolase: FBA). These genes show robust monophyly with those of the marine Prasinophyceae algae genera Micromonas, Ostreococcus and Bathycoccus, indicating that the acquisition of this gene through horizontal gene transfer by an ancestor of the green algal lineage occurred prior to the divergence of the core chlorophytes (Chlorophyceae and Trebouxiophyceae) and the prasinophytes. The absence of this gene in some freshwater chlorophytes, such as Chlamydomonas reinhardtii, Volvox carteri, Chlorella vulgaris, Chlorella variabilis and Coccomyxa subellipsoidea, can therefore be explained by the loss of this gene somewhere in the evolutionary process. Our survey on the distribution of this gene in genomic and transcriptome databases suggests that this gene occurs almost exclusively in marine algae, with a few exceptions, and as such, we propose that chloroplastic class IIA FBA is a marine environment-adapted enzyme. This hypothesis was also experimentally tested using Chlamydomonas W80, for which we found that the transcript levels of this gene to be significantly lower under low-salt (that is, simulated terrestrial) conditions. Expression analyses of transcriptome data for two algae, Prymnesium parvum and Emiliania huxleyi, taken from the Sequence Read Archive database also indicated that the expression of this gene under terrestrial conditions (low NaCl and low sulfate) is significantly downregulated. Thus, these experimental and transcriptome data provide support for our hypothesis. PMID:27058504

Protein (Viridiplantae): 232868 [PGDBj - Ortholog DB

Lifescience Database Archive (English)

Full Text Available 3051:4703 ... 3052:4703 ... 3055:4703 ... hypothetical protein CHLREDRAFT_120274, partial Chlamydomonas reinhardtii PPGCRCSSAPPGCRC...SSAPPGCRCSSAPPGCRCSSAPPGCRCSSAPPGCRCSSAPPGCRCSSAPPGCRCSSAPPGCRCSSAPPGCRCSSAPPGCRCSSAPPGCRCSSAPPGCRCS

Adaptation of light-harvesting functions of unicellular green algae to different light qualities.

Science.gov (United States)

Ueno, Yoshifumi; Aikawa, Shimpei; Kondo, Akihiko; Akimoto, Seiji

2018-05-28

Oxygenic photosynthetic organisms perform photosynthesis efficiently by distributing captured light energy to photosystems (PSs) at an appropriate balance. Maintaining photosynthetic efficiency under changing light conditions requires modification of light-harvesting and energy-transfer processes. In the current study, we examined how green algae regulate their light-harvesting functions in response to different light qualities. We measured low-temperature time-resolved fluorescence spectra of unicellular green algae Chlamydomonas reinhardtii and Chlorella variabilis cells grown under different light qualities. By observing the delayed fluorescence spectra, we demonstrated that both types of green algae primarily modified the associations between light-harvesting chlorophyll protein complexes (LHCs) and PSs (PSII and PSI). Under blue light, Chlamydomonas transferred more energy from LHC to chlorophyll (Chl) located far from the PSII reaction center, while energy was transferred from LHC to PSI via different energy-transfer pathways in Chlorella. Under green light, both green algae exhibited enhanced energy transfer from LHCs to both PSs. Red light induced fluorescence quenching within PSs in Chlamydomonas and LHCs in Chlorella. In Chlorella, energy transfer from PSII to PSI appears to play an important role in balancing excitation between PSII and PSI.

Selenite -Se(4)- uptake mechanisms in the unicellular green alga Chlamydomonas reinhardtii: bioaccumulation and effects induced on growth and ultrastructure; Mecanismes de prise en charge du selenite - Se(4)-chez l'algue verte unicellulaire Chlamydomonas reinhardtii. Bioaccumulation et effets induits sur la croissance et l'ultrastructure

Energy Technology Data Exchange (ETDEWEB)

Morlon, H

2005-03-15

Selenium is an essential element, but becomes very toxic at higher concentrations. It occurs in the environment at concentrations ranging from nM to {mu}M and selenium pollution is a worldwide phenomenon. This works aims at improving the knowledge on the interactions between selenite - Se(IV) - and a freshwater phyto-planktonic organism: the unicellular green algae Chlamydomonas reinhardtii. The aim of the performed experiments were: i) to investigate selenite -Se(IV)- uptake mechanisms in C. reinhardtii, using Se{sup 75} as a tracer in short term exposures (<1 h); ii) to assess selenite toxicity as measured with growth impairment and ultrastructural damage (with EDAX-TEM analysis), using long term exposures (96 h) to stable selenite; iii) to evaluate the bioaccumulation capacity of selenite and its potential links with toxicity. Short-term experiments revealed a negligible adsorption and a time-dependent linear absorption with an estimated absorbed flux of about 0.2 nmol.m{sup -2}.nM{sup -1}.h{sup -1}. The uptake was proportional to ambient levels in a broad range of intermediate concentrations (from nM to {mu}M). However, fluxes were higher at very low concentrations (< nM), and decrease with increasing high concentrations ( > {mu}M), suggesting that a high affinity but rapidly saturated transport mechanism could be used at low concentrations, in parallel with a low affinity mechanism that would only saturate at high concentrations ({approx}mM). The latter could involve transporters used by sulphate and nitrates, as suggested by the inhibition of selenite uptake by those element. Se(IV) speciation changes with pH did not induce significant effect on bioavailability. On the basis of the relationship between Se concentration and maximal cell density achieved, an EC50 of 80 {mu}M ([64; 98]) was derived. No adaptation mechanism were observed as the same the same toxicity was quantified for Se-pre-exposed algae. Observations by TEM suggested chloroplasts as the first

Centrioles are freed from cilia by severing prior to mitosis.

Science.gov (United States)

Parker, Jeremy D K; Hilton, Laura K; Diener, Dennis R; Rasi, M Qasim; Mahjoub, Moe R; Rosenbaum, Joel L; Quarmby, Lynne M

2010-07-01

Cilia are necessary for normal tissue development and homeostasis and are generally present during interphase, but not in mitosis. The precise mechanism of premitotic ciliary loss has been controversial, with data supporting either sequential disassembly through the transition zone or, alternatively, a severing event at the base of the cilia. Here we show by live cell imaging and immunofluorescence microscopy that resorbing flagella of Chlamydomonas leave remnants associated with the mother cell wall. We postulated that the remnants are the product of severing of doublet microtubules between the basal bodies and the flagellar transition zone, thereby freeing the centrioles to participate in spindle organization. We show via TEM that flagellar remnants are indeed flagellar transition zones encased in vesicles derived from the flagellar membrane. This transition zone vesicle can be lodged within the cell wall or it can be expelled into the environment. This process is observable in Chlamydomonas, first because the released flagellar remnants can remain associated with the cell by virtue of attachments to the cell wall, and second because the Chlamydomonas transition zone is particularly rich with electron-dense structure. However, release of basal bodies for spindle-associated function is likely to be conserved among the eukaryotes. 2010 Wiley-Liss, Inc.

Production of terpenes in the culture of Chlorophyceae and Rhodophyta

Science.gov (United States)

Abe, M.; Hashimoto, S.

2014-12-01

Terpenes show high reactivity in the troposphere, contributing to organic aerosol reactions with OH radicals. One of the main sources of terpenes in the atmosphere is terrestrial plants. It has been recently reported that marine phytoplankton also produce monoterpenes (Yassaa et al: 2008). Because aerosol production of natural origin affects the cloud cover over the open ocean, it is important to investigate the origin of aerosol generation in the open ocean. In this study, we investigated the production of terpenes and isoprene with a focus on Chlamydomonas (Chlorophyceae) and Rhodella maculata (Rhodophyta). Concentrations of terpenes and isoprene were measured using a dynamic headspace (GERSTEL DHS)—gas chromatograph (Agilent 6890N)—mass spectrometer (Agilent 5975C). In addition, chlorophyll a was measured using a fluorometer (Turner TD-700). The results showed that isoprene, α-pinene, and β-pinene were produced by Chlamydomonas sp. and that isoprene, limonene, and camphene were produced by Rhodella maculata. Chlamydomonas sp. produced α-pinene and β-pinene, similar to land plants. The ratio of the pinene/isoprene concentrations in the atmosphere over seawater where phytoplankton are blooming has been reported as approximately 0.7 (Yassaa et al: 2008). In this experiment, the pinene/isoprene concentration ratios in the cultures were approximately 0.1. This result indicates that marine phytoplankton may not be ignored in the marine atmosphere chemistry of terpenes.

Carbon and hydrogen matabolism of green algae in light and dark: Final report

Energy Technology Data Exchange (ETDEWEB)

Gibbs, M. [Brandeis Univ., Waltham, MA (United States), Department of Biology

1996-12-31

This report provides an overview of the progress made during this study. Progress is reported in chloroplast respiration, photoregulation of chloroplast respiration, reductive carboxylic acid cycle, and in oxy-hydrogen reaction all in Chlamydomonas.

Generation and characterization of pigment mutants of ...

African Journals Online (AJOL)

acer

One of the most serious ecological problems is muta- ... UV irradiation mutagenesis of Chlamydomonas reinhardtii CC-. 124 .... certain balance between the pigment content in the algal ... is bombarded with the full brunt of solar UV (ultraviolet).

Influence of agglomeration of cerium oxide nanoparticles and speciation of cerium(III) on short term effects to the green algae Chlamydomonas reinhardtii

Energy Technology Data Exchange (ETDEWEB)

Röhder, Lena A. [Eawag, Swiss Federal Institute of Aquatic Science and Technology, Department of Environmental Toxicology, Dübendorf 8600 (Switzerland); ETH-Zurich, Institute of Biogeochemistry and Pollutant Dynamics, Zürich 8092 (Switzerland); Brandt, Tanja [Eawag, Swiss Federal Institute of Aquatic Science and Technology, Department of Environmental Toxicology, Dübendorf 8600 (Switzerland); Sigg, Laura [Eawag, Swiss Federal Institute of Aquatic Science and Technology, Department of Environmental Toxicology, Dübendorf 8600 (Switzerland); ETH-Zurich, Institute of Biogeochemistry and Pollutant Dynamics, Zürich 8092 (Switzerland); Behra, Renata, E-mail: Renata.behra@eawag.ch [Eawag, Swiss Federal Institute of Aquatic Science and Technology, Department of Environmental Toxicology, Dübendorf 8600 (Switzerland)

2014-07-01

Highlights: • Phosphate-dispersed CeO₂ NP did not affect photosynthetic yield in C. reinhardtii. • Agglomerated CeO₂ NP slightly decreased photosynthetic yield. • Cerium(III) was shown to affect photosynthetic yield and intracellular ROS level. • Slight effects of CeO₂ NP were caused by dissolved Ce³⁺ ions present in suspensions. • Wild type and cell wall free mutant of C. reinhardtii showed the same sensitivity. - Abstract: Cerium oxide nanoparticles (CeO₂ NP) are increasingly used in industrial applications and may be released to the aquatic environment. The fate of CeO₂ NP and effects on algae are largely unknown. In this study, the short term effects of CeO₂ NP in two different agglomeration states on the green algae Chlamydomonas reinhardtii were examined. The role of dissolved cerium(III) on toxicity, its speciation and the dissolution of CeO₂ NP were considered. The role of cell wall of C. reinhardtii as a barrier and its influence on the sensitivity to CeO₂ NP and cerium(III) was evaluated by testing both, the wild type and the cell wall free mutant of C. reinhardtii. Characterization showed that CeO₂ NP had a surface charge of ~0 mV at physiological pH and agglomerated in exposure media. Phosphate stabilized CeO₂ NP at pH 7.5 over 24 h. This effect was exploited to test CeO₂ NP dispersed in phosphate with a mean size of 140 nm and agglomerated in absence of phosphate with a mean size of 2000 nm. The level of dissolved cerium(III) in CeO₂ NP suspensions was very low and between 0.1 and 27 nM in all tested media. Exposure of C. reinhardtii to Ce(NO₃)₃ decreased the photosynthetic yield in a concentration dependent manner with EC₅₀ of 7.5 ± 0.84 μM for wild type and EC₅₀ of 6.3 ± 0.53 μM for the cell wall free mutant. The intracellular level of reactive oxygen species (ROS) increased upon exposure to Ce(NO₃)₃ with effective concentrations similar to those inhibiting photosynthesis. The agglomerated Ce

New records of snow algae from the Andes of Ecuador

Czech Academy of Sciences Publication Activity Database

Nedbalová, Linda; Sklenář, P.

2008-01-01

Roč. 15, č. 1 (2008), s. 17-20 ISSN 1815-8242 R&D Projects: GA AV ČR IAA601110702 Institutional research plan: CEZ:AV0Z60050516 Keywords : Chlamydomonas * cryophilous * Cylindrocystis Subject RIV: EF - Botanics

Negative effects of UVB-irradiated phytoplankton on life history traits and fitness of Daphnia magna

NARCIS (Netherlands)

Lange, de H.J.; Reeuwijk, van P.L.

2003-01-01

1. We tested the effect of ultraviolet-B (UVB)-irradiated phytoplankton on life history characteristics of Daphnia magna . Two phytoplankton species were used, Chlamydomonas reinhardtii and Cryptomonas pyrenoidifera . The phytoplankton species were cultured under photosynthetically active radiation

(Carbon and hydrogen metabolism of green algae in light and dark)

Energy Technology Data Exchange (ETDEWEB)

1990-01-01

The focus of this project was the elucidation of anaerobic metabolism in ecuaryotic green algae, chlamydomonas reinhardii. Chlamydomonas is a versatile organism that can grow under disparate conditions such as fresh water lakes and sewage ponds. The cell an photoassimilate CO{sub 2} aerobically and anaerobically, the latter after adaptation'' to a hydrogen metabolism. It can recall the knallgas or oxyhydrogen reaction and utilize hydrogen the simplest of all reducing agents for the dark assimilation of CO{sub 2} by the photosynthetic carbon reduction cycle. The dark reduction with hydrogen lies on the border line between autotrophic and heterotrophic carbon assimilation. Both autotrophic and heterotrophic bacteria are known in which molecular hydrogen can replace either inorganic or organic hydrogen donors. Here the dark reduction of CO{sub 2} acquires a particular importance since it occurs in the same cell that carries on photoreduction and photosynthesis. We will demonstrate here that the alga chloroplast possesses a respiratory capacity. It seems likely that Chlamydomonas may have retained the chloroplastic respiratory pathway because of the selective advantage provided to the algae under a wide range of environmental conditions that the cells experience in nature. The ability to cycle electrons and poise the reduction level of the photosynthetic apparatus under aerobic and microaerobic conditions could allow more efficient CO{sub 2} fixation and enhanced growth under unfavorable conditions or survival under more severe conditions.

[Molecular and structural-biological analysis of Nicotiana plumbaginifolia mutants for identification of the site of beta-tubulins interaction with dinitroanilines and phosphorotioamidates].

Science.gov (United States)

Emets, A I; Baiard, U V; Nyporko, A Iu; Swire-Clark, G A; Blium, Ia B

2009-01-01

The identification of point mutation locations on beta-tubulin molecules of amiprophosmethyl- and trifluralin-resistant Nicotiana plumbaginifolia lines have described in the work. It was shown that in the first case this mutation is connected with the substitution ofserine residue on proline in position 248; in the second case--with the substitution of phenilalanine on serine in position 317 of beta-tubulin amino acid sequence. Three-dimensional models of beta-tubulin molecule from Chlamydomonas with well-known location of mutations conferring dinitroaniline- and phosphorotioamidate resistance (substitution of lysine residue to methionine on position 350), and beta-tubulin from Nicotiana plumbaginifolia have been reconstructed. On the basis of analysis of site of interaction with dinitroanilines and phosphorotioamides on Chlamydomonas beta-tubulin molecule it was concluded that the revealed mutations on Nicotiana plumbaginifolia beta-tubulin affect amino acid residues participating in formation of this site.

Effects of UV-B irradiated algae on life history traits of Daphnia pulex

NARCIS (Netherlands)

De Lange, H.J.; Van Donk, E.

1997-01-01

1. The impact of ultraviolet-B (UVB)-irradiated phytoplankton on the life history parameters of Daphnia was studied. Three species of Chlorophyceae (Chlamydomonas reinhardtii, Scenedesmus acutus and S. subspicatus) and one species of Cryptophyceae (Cryptamonas pyrenoidifera) were cultured with and

Effects of UV-B irradiated algae on zooplankton grazing

NARCIS (Netherlands)

Lange, de H.J.; Lürling, M.F.L.L.W.

2003-01-01

We tested the effects of UV-B stressed algae on grazing rates of zooplankton. Four algal species ( Chlamydomonas reinhardtii, Cryptomonas sp., Scenedesmus obliquus and Microcystis aeruginosa) were used as food and fed to three zooplankton species ( Daphnia galeata, Bosmina longirostris and

Magnetotactic algae

International Nuclear Information System (INIS)

Barros, H.G. de P.L. de; Esquivel, D.M.S.; Danon, J.

1981-01-01

The first observation is reported of an enkaryote micro-organism (chlamydomona), collected in samples from the Rodrigo de Freitas lagune in Rio de Janeiro, which responds to the magnetic field in a similar way as the magnetotactic bacterias. (L.C.) [pt

Degradation and de novo synthesis of D1 protein and psbA ...

Indian Academy of Sciences (India)

This shows that synthesis of D1 protein is not the only component involved in the recovery process. Our events, which ... transcript levels in the green alga Chlamydomonas reinhardtii in ..... and Gaba V 1996 Accelerated degradation of the D2 ...

Transgene expression in microalgae – from tools to applications

Directory of Open Access Journals (Sweden)

Lior eDoron

2016-04-01

Full Text Available Microalgae comprise a biodiverse group of photosynthetic organisms that reside in water sources and sediments. The green microalgae Chlamydomonas reinhardtii was adopted as a useful model organism for studying various physiological systems. Its ability to grow under both photosynthetic and heterotrophic conditions allows efficient growth of non-photosynthetic mutants, making Chlamydomonas a useful genetic tool to study photosynthesis. In addition, this green alga can grow as haploid or diploid cells, similar to yeast, providing a powerful genetic system. As a result, easy and efficient transformation systems have been developed for Chlamydomonas, targeting both the chloroplast and nuclear genomes. Since microalgae comprise a rich repertoire of species that offer variable advantages for biotech and biomed industries, gene transfer technologies were further developed for many microalgae to allow for the expression of foreign proteins of interest. Expressing foreign genes in the chloroplast enables the targeting of foreign DNA to specific sites by homologous recombination. Chloroplast transformation also allows for the introduction of genes encoding several enzymes from a complex pathway, possibly as an operon. Expressing foreign proteins in the chloroplast can also be achieved by introducing the target gene into the nuclear genome, with the protein product bearing a targeting signal that directs import of the transgene-product into the chloroplast, like other endogenous chloroplast proteins. Integration of foreign genes into the nuclear genome is mostly random, resulting in large variability between different clones, such that extensive screening is required. The use of different selection modalities is also described, with special emphasis on the use of herbicides and metabolic markers which are considered to be friendly to the environment, as compared to drug-resistance genes that are commonly used. Finally, despite the development of a wide

The complete chloroplast genome sequence of the chlorophycean green alga Scenedesmus obliquus reveals a compact gene organization and a biased distribution of genes on the two DNA strands

Science.gov (United States)

de Cambiaire, Jean-Charles; Otis, Christian; Lemieux, Claude; Turmel, Monique

2006-01-01

Background The phylum Chlorophyta contains the majority of the green algae and is divided into four classes. While the basal position of the Prasinophyceae is well established, the divergence order of the Ulvophyceae, Trebouxiophyceae and Chlorophyceae (UTC) remains uncertain. The five complete chloroplast DNA (cpDNA) sequences currently available for representatives of these classes display considerable variability in overall structure, gene content, gene density, intron content and gene order. Among these genomes, that of the chlorophycean green alga Chlamydomonas reinhardtii has retained the least ancestral features. The two single-copy regions, which are separated from one another by the large inverted repeat (IR), have similar sizes, rather than unequal sizes, and differ radically in both gene contents and gene organizations relative to the single-copy regions of prasinophyte and ulvophyte cpDNAs. To gain insights into the various changes that underwent the chloroplast genome during the evolution of chlorophycean green algae, we have sequenced the cpDNA of Scenedesmus obliquus, a member of a distinct chlorophycean lineage. Results The 161,452 bp IR-containing genome of Scenedesmus features single-copy regions of similar sizes, encodes 96 genes, i.e. only two additional genes (infA and rpl12) relative to its Chlamydomonas homologue and contains seven group I and two group II introns. It is clearly more compact than the four UTC algal cpDNAs that have been examined so far, displays the lowest proportion of short repeats among these algae and shows a stronger bias in clustering of genes on the same DNA strand compared to Chlamydomonas cpDNA. Like the latter genome, Scenedesmus cpDNA displays only a few ancestral gene clusters. The two chlorophycean genomes share 11 gene clusters that are not found in previously sequenced trebouxiophyte and ulvophyte cpDNAs as well as a few genes that have an unusual structure; however, their single-copy regions differ

Local repeat sequence organization of an intergenic spacer in the ...

Indian Academy of Sciences (India)

Unknown

chloroplast genome of Chlamydomonas reinhardtii leads to DNA expansion and sequence ... The discovery of uniparentally inherited streptomycin resistant mutants ... resembles yeast, mitochondrial and phage recombination in that it is typically ...... Sager R and Lane D 1972 Molecular basis of maternal inheritance; Proc.

Reference: MRNASTA2CRPSBD [PLACE

Lifescience Database Archive (English)

Full Text Available MRNASTA2CRPSBD Nickelsen J, Fleischmann M, Boudreau E, Rahire M, Rochaix JD Identif...ication of cis-acting RNA leader elements required for chloroplast psbD gene expression in Chlamydomonas Plant Cell 11:957-970 (1999) PubMed: 10330479; ...

Evolution of the Phosphatidylcholine Biosynthesis Pathways in Green Algae: Combinatorial Diversity of Methyltransferases.

Science.gov (United States)

Hirashima, Takashi; Toyoshima, Masakazu; Moriyama, Takashi; Sato, Naoki

2018-01-01

Phosphatidylcholine (PC) is one of the most common phospholipids in eukaryotes, although some green algae such as Chlamydomonas reinhardtii are known to lack PC. Recently, we detected PC in four species in the genus Chlamydomonas: C. applanata NIES-2202, C. asymmetrica NIES-2207, C. debaryana NIES-2212, and C. sphaeroides NIES-2242. To reveal the PC biosynthesis pathways in green algae and the evolutionary scenario involved in their diversity, we analyzed the PC biosynthesis genes in these four algae using draft genome sequences. Homology searches suggested that PC in these species is synthesized by phosphoethanolamine-N-methyltransferase (PEAMT) and/or phosphatidylethanolamine-N-methyltransferase (PEMT), both of which are absent in C. reinhardtii. Recombinant PEAMTs from these algae showed methyltransferase activity for phosphoethanolamine but not for monomethyl phosphoethanolamine in vitro, in contrast to land plant PEAMT, which catalyzes the three methylations from phosphoethanolamine to phosphocholine. This suggested an involvement of other methyltransferases in PC biosynthesis. Here, we characterized the putative phospholipid-N-methyltransferase (PLMT) genes of these species by genetic and phylogenetic analysis. Complementation assays using a PC biosynthesis-deficient yeast suggested that the PLMTs of these algae can synthesize PC from phosphatidylethanolamine. These results indicated that the PC biosynthesis pathways in green algae differ from those of land plants, although the enzymes involved are homologous. Phylogenetic analysis suggested that the PEAMTs and PLMTs in these algae were inherited from the common ancestor of green algae. The absence of PC biosynthesis in many Chlamydomonas species is likely a result of parallel losses of PEAMT and PLMT in this genus.

Growth limitation of three Arctic sea-ice algae species: effects of salinitty, pH and inorganic carbon availability

DEFF Research Database (Denmark)

Søgaard, Dorte Haubjerg; Hansen, Per Juel; Rysgaard, Søren

2011-01-01

The effect of salinity, pH, and dissolved inorganic carbon (TCO(2)) on growth and survival of three Arctic sea ice algal species, two diatoms (Fragilariopsis nana and Fragilariopsis sp.), and one species of chlorophyte (Chlamydomonas sp.) was assessed in controlled laboratory experiments. Our res...

Synergistic Effects of Different Food Species on Life-History Traits of Daphnia-Galeata

NARCIS (Netherlands)

Boersma, M.; Vijverberg, J.

1995-01-01

In this paper we describe the life history consequences of feeding Daphnia galeata with different food types in different concentrations. We fed the animals with four concentrations of two green algae Scenedesmus obliquus and Chlamydomonas globosa, given separately as well as in a 1:1 mixture.

On the origin of differentiation

Indian Academy of Sciences (India)

Unknown

a result of an adhesion mutation in a Chlamydomonas- like ancestor. For cellular slime moulds it could mean that gathering together cysts or spores in the soil is advantageous for dispersal and therefore encouraged by selection, and ultimately leading to aggregation by che- motaxis (Bonner 2003). Once multicellularity has ...

Environmental, genetic and cellular toxicity of tenuazonic acid ...

African Journals Online (AJOL)

Alternaria alternata, an important pathogen of many plants, produces tenuazonic acid (TeA) with bioactivity to microbes, plants and animals. TeA is one of the main mycotoxin to humans and other organisms. Using Chlamydomonas reinhardtii, Vicia faba root tip and three mammalian normal cell lines as target materials, ...

Assembly of primary cilia

DEFF Research Database (Denmark)

Pedersen, Lotte B; Veland, Iben R; Schrøder, Jacob M

2008-01-01

knowledge about IFT is based on studies performed in Chlamydomonas and Caenorhabditis elegans. Therefore, our review of the IFT literature includes studies performed in these two model organisms. The role of several non-IFT proteins (e.g., centrosomal proteins) in the ciliary assembly process is also...... discussed. Developmental Dynamics, 2008. (c) 2008 Wiley-Liss, Inc....

Growth of soil algae and cyanobacteria on gold mine tailings material

Directory of Open Access Journals (Sweden)

Tanya Seiderer

2017-11-01

Full Text Available The goal of revegetation of gold mine tailings storage facilities is to reduce aeolian pollution, nutrient leaching and erosion caused by exposure to wind and water. The establishment of biological soil crusts may prove to be a more cost-effective way to reach the same goal and the aim of this study was therefore to determine if it is possible to establish algae and cyanobacteria on gold mine tailings. Different treatments of Chlamydomonas, Microcoleus and Nostoc were inoculated on gold mine tailings in controlled conditions and algal growth was measured on all of the treatments after 6 weeks. Nostoc treatments had the highest chlorophyll-a concentrations and produced a surface crust, while Chlamydomonas treatments penetrated the tailings material and provided the strongest crust. The results were promising but more research is necessary to determine the best organism, or combination of organisms, to colonise mine tailings and to eventually produce biological crusts. Significance: Determination of the best organisms to colonise mine tailings and to produce biological crusts for the revegetation of gold mine tailings storage facilities.

Cultivation of micro-algae in closed tubular reactor

Energy Technology Data Exchange (ETDEWEB)

Gudin, C.; Bernard, A.; Chaumont, D.

1983-11-01

A description is presented of the three culture pilot utilities in activity under natural light, including glass tubular solar collector (30 mm diameter) in which the microalgae culture circulates. The utility is controled automatically (thermal regulation, gaseous transfers, continuous culture organization). The tests were conducted for the production of polysaccharides (Porphyridium cruentum, chlamydomonas mexicana) or hydrocarbons (Botriococcus braunii).

Bioaccumulation and toxicity of selenium compounds in the green alga Scenedesmus quadricauda

Czech Academy of Sciences Publication Activity Database

Umysová, Dáša; Vítová, Milada; Doušková, Irena; Bišová, Kateřina; Hlavová, Monika; Čížková, Mária; Machat, J.; Doucha, Jiří; Zachleder, Vilém

2009-01-01

Roč. 9, č. 58 (2009), s. 1-16 ISSN 1471-2229 R&D Projects: GA AV ČR IAA600200701; GA MŠk OE 221; GA MŠk OE09025 Institutional research plan: CEZ:AV0Z50200510 Keywords : CHLAMYDOMONAS-REINHARDTII * THIOREDOXIN REDUCTASE * EMILIANIA-HUXLEYI Subject RIV: EE - Microbiology, Virology Impact factor: 3.774, year: 2009

A universal protocol for the combined isolation of metabolites, DNA, long RNAs, small RNAs, and proteins from plants and microorganisms

Czech Academy of Sciences Publication Activity Database

Valledor, Luis; Escandón, M.; Meijón, M.; Nukarinen, E.; Jesús Cañal, M.; Weckwerth, W.

2014-01-01

Roč. 79, č. 1 (2014), s. 173-180 ISSN 0960-7412 R&D Projects: GA MŠk(CZ) EE2.3.20.0256 Institutional support: RVO:67179843 Keywords : systems biology * combined isolation * RNA * small RNA * proteins * metabolites * Chlamydomonas reinhardtii * Arabidopsis thaliana * Populus sp. * Pinus sp. * technical advance Subject RIV: EI - Biotechnology ; Bionics Impact factor: 5.972, year: 2014

Alga-based HPV16 E7 vaccine elicits specific immune response in mice

Czech Academy of Sciences Publication Activity Database

Vlasák, Josef; Bříza, Jindřich; Ryba, Š.; Ludvíková, V.

2013-01-01

Roč. 34, č. 1 (2013), s. 141-148 ISSN 2249-7412 R&D Projects: GA AV ČR IAA500960903 Institutional support: RVO:60077344 Keywords : Chlamydomonas reinhardtii * chloroplast transformation * human papillomaviruses * E7 oncogene Subject RIV: EB - Genetics ; Molecular Biology http://pelagiaresearchlibrary.com/asian-journal-of-plant-science/vol3-iss1/AJPSR-2013-3-1-141-148.pdf

Phosphopantetheinylation in the green microalgae Chlamydomonas reinhardtii

DEFF Research Database (Denmark)

Sonnenschein, Eva; Pu, Yuan; Beld, Joris

2016-01-01

available microalgal genome data revealed that most green microalgae appear to carry two PPTases forming clusters with each C. reinhardtii PPTase, while microalgae of other divisions carry one or two PPTases and do not cluster in the pattern of the green algal data. This new understanding on the PPTases...... in microalgae shows that microalgae are already primed for biotechnological applications in contrast to other organisms. Thus, microalgae have great potential for metabolic engineering efforts in the realm of biofuel and high-value products including direct engineering of the fatty acid or secondary metabolism...

Role of the Rubisco small subunit. Final report for period May 1, 1997--April 30,2000

Energy Technology Data Exchange (ETDEWEB)

Spreitzer, Robert J.

2000-10-04

CO{sub 2} and O{sub 2} are mutually competitive at the active site of ribulose-1,5-biphosphate (RuBP) carboxylase/oxygenase (Rubisco). Rubisco contains two subunits, each present in eight copies. The 15-kD small subunit is coded by a family of nuclear RbcS genes. Until now, the role of the small subunit in Rubisco structure or catalytic efficiency is not known. Because of other work in eliminating the two RbcS genes in the green algo Chlamydomonas reinhardtii, it is now possible to address questions about the structure-function relationships of the eukaryotic small subunit. There are three specific aims in this project: (1) Alanine scanning mutagenesis is being used to dissect the importance of the {beta}A/{beta}B loop, a feature unique to the eukaryotic small subunit. (2) Random mutagenesis is being used to identify additional residues or regions of the small subunit that are important for holoenzyme assembly and function. (3) Attempts are being made to express foreign small subunits in Chlamydomonas to examine the contribution of small subunits to holoenzyme assembly, catalytic efficiency, and CO{sub 2}/O{sub 2} specificity.

Cryptochrome photoreceptors in green algae: Unexpected versatility of mechanisms and functions.

Science.gov (United States)

Kottke, Tilman; Oldemeyer, Sabine; Wenzel, Sandra; Zou, Yong; Mittag, Maria

2017-10-01

Green algae have a highly complex and diverse set of cryptochrome photoreceptor candidates including members of the following subfamilies: plant, plant-like, animal-like, DASH and cryptochrome photolyase family 1 (CPF1). While some green algae encode most or all of them, others lack certain members. Here we present an overview about functional analyses of so far investigated cryptochrome photoreceptors from the green algae Chlamydomonas reinhardtii (plant and animal-like cryptochromes) and Ostreococcus tauri (CPF1) with regard to their biological significance and spectroscopic properties. Cryptochromes of both algae have been demonstrated recently to be involved to various extents in circadian clock regulation and in Chlamydomonas additionally in life cycle control. Moreover, CPF1 even performs light-driven DNA repair. The plant cryptochrome and CPF1 are UVA/blue light receptors, whereas the animal-like cryptochrome responds to almost the whole visible spectrum including red light. Accordingly, plant cryptochrome, animal-like cryptochrome and CPF1 differ fundamentally in their structural response to light as revealed by their visible and infrared spectroscopic signatures, and in the role of the flavin neutral radical acting as dark form or signaling state. Copyright © 2017 Elsevier GmbH. All rights reserved.

Evolution of an atypical de-epoxidase for photoprotection in the green lineage.

Science.gov (United States)

Li, Zhirong; Peers, Graham; Dent, Rachel M; Bai, Yong; Yang, Scarlett Y; Apel, Wiebke; Leonelli, Lauriebeth; Niyogi, Krishna K

2016-09-12

Plants, algae and cyanobacteria need to regulate photosynthetic light harvesting in response to the constantly changing light environment. Rapid adjustments are required to maintain fitness because of a trade-off between efficient solar energy conversion and photoprotection. The xanthophyll cycle, in which the carotenoid pigment violaxanthin is reversibly converted into zeaxanthin, is ubiquitous among green algae and plants and is necessary for the regulation of light harvesting, protection from oxidative stress and adaptation to different light conditions(1,2). Violaxanthin de-epoxidase (VDE) is the key enzyme responsible for zeaxanthin synthesis from violaxanthin under excess light. Here we show that the Chlorophycean VDE (CVDE) gene from the model green alga Chlamydomonas reinhardtii encodes an atypical VDE. This protein is not homologous to the VDE found in plants and is instead related to a lycopene cyclase from photosynthetic bacteria(3). Unlike the plant-type VDE that is located in the thylakoid lumen, the Chlamydomonas CVDE protein is located on the stromal side of the thylakoid membrane. Phylogenetic analysis suggests that CVDE evolved from an ancient de-epoxidase that was present in the common ancestor of green algae and plants, providing evidence of unexpected diversity in photoprotection in the green lineage.

Development of an efficient algal H{sub 2}-producing system

Energy Technology Data Exchange (ETDEWEB)

Ghirardi, M.L.; Markov, S.; Seibert, M. [National Renewable Energy Lab., Golden, CO (United States)

1996-10-01

Green algae have the potential to efficiently photoevolve H{sub 2} from water using the photosynthetic O{sub 2} evolving apparatus and the reversible hydrogenase enzyme when CO{sub 2} is not present. Unfortunately algal hydrogenases are very sensitive to inactivation by O{sub 2}, the by-product of the water-splitting process. This problem has been one of the major practical factors limiting the commercial utilization of green algae for H{sub 2} production. The other major limitation, saturation of H{sub 2} production by algae at light intensities much lower than normal solar levels, is being addressed by ORNL. The objectives of this project are to generate O{sub 2}-tolerant, H{sub 2}-producing mutants of the green alga Chlamydomonas reinhardtti, to test them in a laboratory-scale system for continuous production of H{sub 2} under aerobic conditions; and to collaborate with ORNL to improve the overall efficiency of H{sub 2} production in intact and cell-free systems. The ultimate goal of the work is to configure a photobiological water-splitting process that will lead to a H{sub 2}-producing system that is cost effective, scalable, non-polluting, and renewable. The approach to obtain O{sub 2}-tolerant mutants of Chlamydomonas involves two types of selection techniques. The first depends on the survival of cells under photoreductive conditions, where H{sub 2} utilization is required, and the second requires the survival of the organisms under H{sub 2}-producing conditions. As part of this collaboration, the authors have independently confirmed that two of the Chlamydomonas mutants lacking photosystem I used by ORNL do in fact produce O{sub 2} in the light and also evolve H{sub 2}. Not unexpectedly, they do the latter with the same O{sub 2}-sensitivity as the WT cells. This observation is crucial for the credibility of the important ORNL work, since it confirms the potential for doubling the quantum efficiency for H{sub 2} production in these mutants.

Flagellar Synchronization Is a Simple Alternative to Cell Cycle Synchronization for Ciliary and Flagellar Studies.

Science.gov (United States)

Dutta, Soumita; Avasthi, Prachee

2017-01-01

The unicellular green alga Chlamydomonas reinhardtii is an ideal model organism for studies of ciliary function and assembly. In assays for biological and biochemical effects of various factors on flagellar structure and function, synchronous culture is advantageous for minimizing variability. Here, we have characterized a method in which 100% synchronization is achieved with respect to flagellar length but not with respect to the cell cycle. The method requires inducing flagellar regeneration by amputation of the entire cell population and limiting regeneration time. This results in a maximally homogeneous distribution of flagellar lengths at 3 h postamputation. We found that time-limiting new protein synthesis during flagellar synchronization limits variability in the unassembled pool of limiting flagellar protein and variability in flagellar length without affecting the range of cell volumes. We also found that long- and short-flagella mutants that regenerate normally require longer and shorter synchronization times, respectively. By minimizing flagellar length variability using a simple method requiring only hours and no changes in media, flagellar synchronization facilitates the detection of small changes in flagellar length resulting from both chemical and genetic perturbations in Chlamydomonas . This method increases our ability to probe the basic biology of ciliary size regulation and related disease etiologies. IMPORTANCE Cilia and flagella are highly conserved antenna-like organelles that found in nearly all mammalian cell types. They perform sensory and motile functions contributing to numerous physiological and developmental processes. Defects in their assembly and function are implicated in a wide range of human diseases ranging from retinal degeneration to cancer. Chlamydomonas reinhardtii is an algal model system for studying mammalian cilium formation and function. Here, we report a simple synchronization method that allows detection of small

Characterization of Gasolines, Diesel Fuels and Their Water Soluble Fractions

Science.gov (United States)

1983-09-01

Hutchinson, et al.,1979 ) with the marine algae, Chlorella vulgaris and Chlamydomonas angulosa, suggests that the toxicity of hydrocarbons is a...water-soluble petroleum components on the growth of Chlorella vulgaris Beijernck. Environ. Poll. 9: 157. Morrow, J.E., et al. 1975. Effects of some...P.B., and T.C. Hutchison. 1975. The effects of water-soluble petroleum components on the growth of Chlorella vulqaris Beijerinck. Environ. Poll. 9

Completion of cell division is associated with maximum telomerase activity in naturally synchronized cultures of the green alga Desmodesmus quadricauda

Czech Academy of Sciences Publication Activity Database

Ševčíková, Tereza; Bišová, Kateřina; Fojtová, Miloslava; Lukešová, Alena; Hrčková, Kristýna; Sýkorová, Eva

2013-01-01

Roč. 587, č. 6 (2013), s. 743-748 ISSN 0014-5793 R&D Projects: GA ČR(CZ) GA521/09/1912; GA MŠk(CZ) ED1.1.00/02.0068 Institutional research plan: CEZ:AV0Z50040702 Institutional support: RVO:68081707 ; RVO:60077344 ; RVO:61388971 Keywords : SCENEDESMUS-QUADRICAUDA * CYCLE * CHLAMYDOMONAS Subject RIV: BO - Biophysics; EE - Microbiology, Virology (MBU-M) Impact factor: 3.341, year: 2013

In the presence of fluoride, free Sc³⁺ is not a good predictor of Sc bioaccumulation by two unicellular algae: possible role of fluoro-complexes.

Science.gov (United States)

Crémazy, Anne; Campbell, Peter G C; Fortin, Claude

2014-08-19

We investigated the effect of fluoride complexation on scandium accumulation by two unicellular algae, Chlamydomonas reinhardtii and Pseudokirchneriella subcapitata. This trivalent metal was selected for its chemical similarities with aluminum and for its convenient radioisotope (Sc-46), which can be used as a tracer in short-term bioaccumulation studies. Scandium surface-bound concentrations (Sc(ads)) and uptake fluxes (J(int)) were estimated in the two algae over short-term (organisms.

Effect of light on respiration and development of photosynthetic cells. Progress report, September 1, 1975--June 1, 1976

Energy Technology Data Exchange (ETDEWEB)

Gibbs, M

1976-06-01

Adaptation and deadaptation of unicellular algal cells to a hydrogen metabolism may involve not only hydrogenase but also the site of entry of electrons from NADH. NADPH can also serve as an electron donor in a chloroplast preparation fortified with hydrogenase but the rate of photoevolution falls off rapidly perhaps due to competition with NADP for electrons. The evolution of H/sub 2/ with reduced pyridine nucleotide as electron donor is coupled to ATP formation. Evidence is presented that soluble ferredoxin may not be the immediate donor of H/sub 2/. Hydrogenases from Chlamydomonas and Scenedesmus have been highly purified and will be used to test the stoichiometry of NADH disappearance, ATP and H/sub 2/ formation and also whether H/sub 2/-photoevolution in contrast to dark evolution is ferredoxin dependent. Studies with intact Chlamydomonas provide evidence that a reduced carbon compound and not water is the source of H/sub 2/. The presence of acetate eliminates the CO/sub 2/ production. The ratio of CO/sub 2/:H/sub 2/ is 1:2 and evolution is blocked by inhibitors of glycolysis. Acetate has a strong effect on H/sub 2/O. Cells starved over a long period still retain photoreduction but will not evolve H/sub 2/ until glucose is added.

A two dimensional clinostat experiment for microalgae cultures - basic work for bio- regenerativ life support systems

Science.gov (United States)

Harting, Benjamin; Slenzka, Klaus

2012-07-01

To investigate the influence of microgravity environments on photosynthetic organisms we designed a 2 dimensional clinostatexperiment for a suspended cell culture of Chlamydomonas reinhardtii. A novel approach of online measurments concerning relevant parameters important for the clasification of photosynthesis was obtained. To adress the photosynthesis rate we installed and validated an optical mesurement system to monitor the evolution and consumption of dissolved oxygen. Simultaneously a PAM sensor to analyse the flourescence quantum yield of the photochemical reaction was integarted. Thus it was possible to directly classify important parameters of the phototrophic metabolism during clinorotation. The experiment design including well suited light conditions and further biochemical analysis were directly performed for microalgal cell cultures. Changes in the photosynthetic efficiancy of phototrophic cyanobacteria has been observed during parabolic flight campaign but the cause is already not understood. Explenations could be the dependency of gravitaxis by intracellular ionconcentartion or the existance of mechanosensitive ionchannels for example associated in chloroplasts of Chlamydomonas reinhardtii. The purpuse of the microalgal clinostat are studies in a qasi microgravity environment for the process design of future bioregenerative life suport systems in spaceflight missions. First results has indicated the need for special nourishment of the cell culture during microgravity experiments. Further data will be presented during the assembly.

Dynein light chain family in Tetrahymena thermophila.

Science.gov (United States)

Wilkes, David E; Rajagopalan, Vidyalakshmi; Chan, Clarence W C; Kniazeva, Ekaterina; Wiedeman, Alice E; Asai, David J

2007-02-01

Dyneins are large protein complexes that produce directed movement on microtubules. In situ, dyneins comprise combinations of heavy, intermediate, light-intermediate, and light chains. The light chains regulate the locations and activities of dyneins but their functions are not completely understood. We have searched the recently sequenced Tetrahymena thermophila macronuclear genome to describe the entire family of dynein light chains expressed in this organism. We identified fourteen genes encoding putative dynein light chains and seven genes encoding light chain-like proteins. RNA-directed PCR revealed that all 21 genes were expressed. Quantitative real time reverse transcription PCR showed that many of these genes were upregulated after deciliation, indicating that these proteins are present in cilia. Using the nomenclature developed in Chlamydomonas, Tetrahymena expresses two isoforms each of LC2, LC4, LC7, and Tctex1, three isoforms of p28, and six LC8/LC8-like isoforms. Tetrahymena also expresses two LC3-like genes. No Tetrahymena orthologue was found for Chlamydomonas LC5 or LC6. This study provides a complete description of the different genes and isoforms of the dynein light chains that are expressed in Tetrahymena, a model organism in which the targeted manipulation of genes is straightforward. (c) 2006 Wiley-Liss, Inc.

変動重力場に対する生物対流時空間パターンの応答

OpenAIRE

Mogami, Yoshihiro; Asato, Eriko; Wada, Yuuko; Kubota, Ayami; Katsu, Yumiko; Niihori, Maki; Baba, Shoji A.; 最上, 善広; 安里, 枝利子; 和田, 祐子; 久保田, 彩水; 勝, 由美子; 新堀, 真希; 馬場, 昭次

2009-01-01

Bioconvection of the aquatic microorganisms has been focused as an experimental tool for the research proposal aiming to reveal the possibility for gravity to develop new functions of the biological systems through the collective interactions between the individual components of the systems. Previously we reported the changes in the bioconvection pattern of Tetrahymena and Chlamydomonas, viewed from top, in response to the altered gravity during parabolic fight by an airplane. In top-view rec...

Photosynthetic efficiency of Chlamydomonas reinhardtii in flashing light

NARCIS (Netherlands)

Vejrazka, C.; Janssen, M.G.J.; Streefland, M.; Wijffels, R.H.

2011-01-01

Efficient light to biomass conversion in photobioreactors is crucial for economically feasible microalgae production processes. It has been suggested that photosynthesis is enhanced in short light path photobioreactors by mixing-induced flashing light regimes. In this study, photosynthetic

Transgene Expression in Microalgae-From Tools to Applications.

Science.gov (United States)

Doron, Lior; Segal, Na'ama; Shapira, Michal

2016-01-01

Microalgae comprise a biodiverse group of photosynthetic organisms that reside in water sources and sediments. The green microalgae Chlamydomonas reinhardtii was adopted as a useful model organism for studying various physiological systems. Its ability to grow under both photosynthetic and heterotrophic conditions allows efficient growth of non-photosynthetic mutants, making Chlamydomonas a useful genetic tool to study photosynthesis. In addition, this green alga can grow as haploid or diploid cells, similar to yeast, providing a powerful genetic system. As a result, easy and efficient transformation systems have been developed for Chlamydomonas, targeting both the chloroplast and nuclear genomes. Since microalgae comprise a rich repertoire of species that offer variable advantages for biotech and biomed industries, gene transfer technologies were further developed for many microalgae to allow for the expression of foreign proteins of interest. Expressing foreign genes in the chloroplast enables the targeting of foreign DNA to specific sites by homologous recombination. Chloroplast transformation also allows for the introduction of genes encoding several enzymes from a complex pathway, possibly as an operon. Expressing foreign proteins in the chloroplast can also be achieved by introducing the target gene into the nuclear genome, with the protein product bearing a targeting signal that directs import of the transgene-product into the chloroplast, like other endogenous chloroplast proteins. Integration of foreign genes into the nuclear genome is mostly random, resulting in large variability between different clones, such that extensive screening is required. The use of different selection modalities is also described, with special emphasis on the use of herbicides and metabolic markers which are considered to be friendly to the environment, as compared to drug-resistance genes that are commonly used. Finally, despite the development of a wide range of transformation

Photoreception in Phytoplankton.

Science.gov (United States)

Colley, Nansi Jo; Nilsson, Dan-Eric

2016-11-01

In many species of phytoplankton, simple photoreceptors monitor ambient lighting. Photoreceptors provide a number of selective advantages including the ability to assess the time of day for circadian rhythms, seasonal changes, and the detection of excessive light intensities and harmful UV light. Photoreceptors also serve as depth gauges in the water column for behaviors such as diurnal vertical migration. Photoreceptors can be organized together with screening pigment into visible eyespots. In a wide variety of motile phytoplankton, including Chlamydomonas, Volvox, Euglena, and Kryptoperidinium, eyespots are light-sensitive organelles residing within the cell. Eyespots are composed of photoreceptor proteins and typically red to orange carotenoid screening pigments. This association of photosensory pigment with screening pigment allows for detection of light directionality, needed for light-guided behaviors such as positive and negative phototaxis. In Chlamydomonas, the eyespot is located in the chloroplast and Chlamydomonas expresses a number of photosensory pigments including the microbial channelrhodopsins (ChR1 and ChR2). Dinoflagellates are unicellular protists that are ecologically important constituents of the phytoplankton. They display a great deal of diversity in morphology, nutritional modes and symbioses, and can be photosynthetic or heterotrophic, feeding on smaller phytoplankton. Dinoflagellates, such as Kryptoperidinium foliaceum, have eyespots that are used for light-mediated tasks including phototaxis. Dinoflagellates belonging to the family Warnowiaceae have a more elaborate eye. Their eye-organelle, called an ocelloid, is a large, elaborate structure consisting of a focusing lens, highly ordered retinal membranes, and a shield of dark pigment. This complex eye-organelle is similar to multicellular camera eyes, such as our own. Unraveling the molecular makeup, structure and function of dinoflagellate eyes, as well as light-guided behaviors in

Diurnal changes in the xanthophyll cycle pigments of freshwater algae correlate with the environmental hydrogen peroxide concentration rather than non-photochemical quenching

Science.gov (United States)

Roach, Thomas; Miller, Ramona; Aigner, Siegfried; Kranner, Ilse

2015-01-01

Background and Aims In photosynthetic organisms exposure to high light induces the production of reactive oxygen species (ROS), such as hydrogen peroxide (H2O2), which in part is prevented by non-photochemical quenching (NPQ). As one of the most stable and longest-lived ROS, H2O2 is involved in key signalling pathways in development and stress responses, although in excess it can induce damage. A ubiquitous response to high light is the induction of the xanthophyll cycle, but its role in algae is unclear as it is not always associated with NPQ induction. The aim of this study was to reveal how diurnal changes in the level of H2O2 are regulated in a freshwater algal community. Methods A natural freshwater community of algae in a temporary rainwater pool was studied, comprising photosynthetic Euglena species, benthic Navicula diatoms, Chlamydomonas and Chlorella species. Diurnal measurements were made of photosynthetic performance, concentrations of photosynthetic pigments and H2O2. The frequently studied model organisms Chlamydomonas and Chlorella species were isolated to study photosynthesis-related H2O2 responses to high light. Key Results NPQ was shown to prevent H2O2 release in Chlamydomonas and Chlorella species under high light; in addition, dissolved organic carbon excited by UV-B radiation was probably responsible for a part of the H2O2 produced in the water column. Concentrations of H2O2 peaked at 2 µm at midday and algae rapidly scavenged H2O2 rather than releasing it. A vertical H2O2 gradient was observed that was lowest next to diatom-rich benthic algal mats. The diurnal changes in photosynthetic pigments included the violaxanthin and diadinoxanthin cycles; the former was induced prior to the latter, but neither was strictly correlated with NPQ. Conclusions The diurnal cycling of H2O2 was apparently modulated by the organisms in this freshwater algal community. Although the community showed flexibility in its levels of NPQ, the diurnal changes in

Polysaccharide-producing microalgae

Energy Technology Data Exchange (ETDEWEB)

Braud, J.P.; Chaumont, D.; Gudin, C.; Thepenier, C.; Chassin, P.; Lemaire, C.

1982-11-01

The production of extracellular polysaccharides is studied with Nostoc sp (cyanophycus), Porphiridium cruentum, Rhodosorus marinus, Rhodella maculata (rhodophyci) and Chlamydomonas mexicana (chlorophycus). The polysaccharides produced are separated by centrifugation of the culture then precipitation with alcohol. Their chemical structure was studied by infrared spectrometry and acid hydrolysis. By their rheological properties and especially their insensitivity to temperatrure and pH variations the polysaccharides produced by Porphryridium cruentum and Rhodella maculata appear as suitable candidates for industrial applications.

The microorganisms as a renewable source of ecological clean fuel

International Nuclear Information System (INIS)

Shalygo, N.V.; Mel'nikov, S.S.; Manankina, E.E.; Budakova, E.A.; Kolyago, V.M.

2006-01-01

Five families of microorganisms (Bacillaceae, Rhodospirillaceae, Cyanophyceae, Chlorophyceae and Euglenophyceae) as hydrogen producers were tested and the conditions that are necessary for hydrogen photoproduction were investigated. It was shown, that the most effective producers of hydrogen were Rhodobacter spheroides, Clostridium sp.; Euglena gracilis var. bacillaris and Chlamydomonas reinhardtii. Addition of glucose, iron and vanadium salts resulted in the increase of hydrogen production. Polycultures consisted of two or three microorganisms were more effective hydrogen producers compared to separate monocultures. (authors)

Quantification of silver nanoparticle toxicity to algae in soil via photosynthetic and flow-cytometric analyses

OpenAIRE

Nam, Sun-Hwa; Il Kwak, Jin; An, Youn-Joo

2018-01-01

Soil algae, which have received attention for their use in a novel bioassay to evaluate soil toxicity, expand the range of terrestrial test species. However, there is no information regarding the toxicity of nanomaterials to soil algae. Thus, we evaluated the effects of silver nanoparticles (0–50 mg AgNPs/kg dry weight soil) on the soil alga Chlamydomonas reinhardtii after six days, and assessed changes in biomass, photosynthetic activity, cellular morphology, membrane permeability, esterase ...

Hyperspectral imaging of snow algae and green algae from aeroterrestrial habitats.

Science.gov (United States)

Holzinger, Andreas; Allen, Michael C; Deheyn, Dimitri D

2016-09-01

Snow algae and green algae living in aeroterrestrial habitats are ideal objects to study adaptation to high light irradiation. Here, we used a detailed description of the spectral properties as a proxy for photo-acclimation/protection in snow algae (Chlamydomonas nivalis, Chlainomonas sp. and Chloromonas sp.) and charophyte green algae (Zygnema sp., Zygogonium ericetorum and Klebsormidium crenulatum). The hyperspectral microscopic mapping and imaging technique allowed us to acquire total absorption spectra of these microalgae in the waveband of 400-900nm. Particularly in Chlamydomonas nivalis and Chlainomonas sp., a high absorbance between 400-550nm was observed, due to naturally occurring secondary carotenoids; in Chloromonas sp. and in the charopyhte algae this high absorbance was missing, the latter being close relatives to land plants. To investigate if cellular water loss has an influence on the spectral properties, the cells were plasmolysed in sorbitol or desiccated at ambient air. While in snow algae, these treatments did hardly change the spectral properties, in the charopyhte algae the condensation of the cytoplasm and plastids increased the absorbance in the lower waveband of 400-500nm. These changes might be ecologically relevant and photoprotective, as aeroterrestrial algae are naturally exposed to occasional water limitation, leading to desiccation, which are conditions usually occurring together with higher irradiation. Copyright © 2016 The Authors. Published by Elsevier B.V. All rights reserved.

Cloning and analysis of calmodulin gene from Porphyra yezoensis Ueda (Bangiales, Rhodophyta)

Science.gov (United States)

Wang, Mengqiang; Mao, Yunxiang; Zhuang, Yunyun; Kong, Fanna; Sui, Zhenghong

2009-09-01

In order to understand the mechanisms of signal transduction and anti-desiccation mechanisms of Porphyra yezoensis, cDNA and its genomic sequence of Calmodulin gene (CaM) was cloned by the technique of polymerase chain reaction (PCR) based on the analysis of P. yezoensis ESTs from dbEST database. The result shows that the full-length cDNA of CaM consists of 603 bps including an ORF encoding for 151 amino acids and a terminate codon UGA, while the length of genomic sequence is 1231 bps including 2 exons and 1 intron. The average GC content of the coding region is 58.77%, while the GC content of the third position of this gene is as high as 82.23%. Four Ca2+ binding sites (EF-hand) are found in this gene. The predicted molecular mass of the deduced peptide is 16688.72 Da and the pI is 4.222. By aligning with known CaM genes, the similarity of CaM gene sequence with homologous genes in Chlamydomonas incerta and Chlamydomonas reinhardtii is 72.7% and 72.2% respectively, and the similarity of the deduced amino acid sequence of CaM gene with homologous genes in C. incerta and C. reinhardtii are both 71.5%. This is the first report on CaM from a species of Rhodophyta.

A Novel Miniature Culture System to Screen CO2-Sequestering Microalgae

Directory of Open Access Journals (Sweden)

Xiaoling Miao

2012-11-01

Full Text Available In this study, a novel 96-well microplate swivel system (M96SS was built for high-throughput screening of microalgal strains for CO2 fixation. Cell growth under different CO2 supply conditions (0.2, 0.4, 0.8, and 1.2 g L−1 d−1, residual nitrate, and pH value of Chlorella sp. SJTU-3, Chlorella pyrenoidosa SJTU-2, and Scenedesmus obliquus SJTU-3 were examined in the M96SS and traditional flask cultures. The dynamic data showed there was a good agreement between the systems. Two critical problems in miniature culture systems (intra-well mixing and evaporation loss were improved by sealed vertical mixing of the M96SS. A sample screen of six microalgal species (Chlorella sp. SJTU-3, Chlorella pyrenoidosa SJTU-2, Selenastrum capricornutum, Scenedesmus obliquus SJTU-3, Chlamydomonas sajao, Dunaliella primolecta was carried out in flasks and the M96SS. Chlamydomonas sajao appeared to be a robust performer (highest cell density: 1.437 g L−1 in anaerobic pond water with 0.8, and 1.2 g L−1 d−1 CO2. The reliability and efficiency of the M96SS were verified through a comparison of traditional flask culture, M96SS, Lukavský’s system, and a microplate shaker.

Enhancement of total lipid yield by nitrogen, carbon, and iron supplementation in isolated microalgae.

Science.gov (United States)

Sivaramakrishnan, Ramachandran; Incharoensakdi, Aran

2017-08-01

The biochemical contents and biodiesel production ability of three microalgal strains grown under different sodium nitrate, sodium carbonate, and ferric ammonium citrate (iron) levels were investigated. The highest biomass and lipid contents were found in Scenedesmus sp., Chlorella sp., and Chlamydomonas sp. when grown in normal BG-11 containing sodium carbonate concentration at 0.03 g · L -1 , and in normal BG-11 containing iron concentration (IC) at 0.009 or 0.012 g · L -1 . Increasing the sodium nitrate level increased the biomass content, but decreased the lipid content in all three microalgae. Among the three microalgae, Scenedesmus sp. showed the highest total lipid yield of 0.69 g · L -1 under the IC of 0.012 g · L -1 . Palmitic and oleic acids were the major fatty acids of Scenedesmus sp. and Chlamydomonas sp. lipids. On the other hand, Chlorella sp. lipids were rich in palmitic, oleic, and linolenic acids, and henceforth contributing to poor biodiesel properties below the standard limits. The three isolated strains had a potential for biodiesel production. Nevertheless, Scenedesmus sp. from stone quarry pond water was the most suitable source for biodiesel production with tolerance toward the high concentration of sodium carbonate without the loss of its biodiesel properties. © 2017 Phycological Society of America.

Calcium sensors of ciliary outer arm dynein: functions and phylogenetic considerations for eukaryotic evolution.

Science.gov (United States)

Inaba, Kazuo

2015-01-01

The motility of eukaryotic cilia and flagella is modulated in response to several extracellular stimuli. Ca(2+) is the most critical intracellular factor for these changes in motility, directly acting on the axonemes and altering flagellar asymmetry. Calaxin is an opisthokont-specific neuronal calcium sensor protein first described in the sperm of the ascidian Ciona intestinalis. It binds to a heavy chain of two-headed outer arm dynein in a Ca(2+)-dependent manner and regulates 'asymmetric' wave propagation at high concentrations of Ca(2+). A Ca(2+)-binding subunit of outer arm dynein in Chlamydomonas reinhardtii, the light chain 4 (LC4), which is a Ca(2+)-sensor phylogenetically different from calaxin, shows Ca(2+)-dependent binding to a heavy chain of three-headed outer arm dynein. However, LC4 appears to participate in 'symmetric' wave propagation at high concentrations of Ca(2+). LC4-type dynein light chain is present in bikonts, except for some subclasses of the Excavata. Thus, flagellar asymmetry-symmetry conversion in response to Ca(2+) concentration represents a 'mirror image' relationship between Ciona and Chlamydomonas. Phylogenetic analyses indicate the duplication, divergence, and loss of heavy chain and Ca(2+)-sensors of outer arm dynein among excavate species. These features imply a divergence point with respect to Ca(2+)-dependent regulation of outer arm dynein in cilia and flagella during the evolution of eukaryotic supergroups.

Genome, Functional Gene Annotation, and Nuclear Transformation of the Heterokont Oleaginous Alga Nannochloropsis oceanica CCMP1779

Science.gov (United States)

2012-11-15

development of such an algal model system for basic discovery, we sequenced the genome and two sets of transcriptomes of N. oceanica CCMP1779, assembled...CCMP1779 has a gene encoding a highly conserved violax- anthin de-epoxidase ( VDE ) protein like that found in plants (Table S9). In Arabidopsis, VDE is...HLA3 or LCI1 were present. This result suggests that CCMP1779 might have a plastid Ci transport system similar to that of Chlamydomonas, but a distinct

Effects of near ultraviolet and green radiations on plant growth

Energy Technology Data Exchange (ETDEWEB)

Klein, R.M.; Edsall, P.C.; Gentile, A.C.

1965-01-01

Selective removal of near ultraviolet and green wavelengths from white light permitted enhanced growth of marigold, tomato, corn, and Impatiens plants, Chlamydomonas cells and the mycelium of Sordaria. Additions of near ultraviolet and green radiations caused repressions in the growth of marigold and Sordaria. These wavelengths do not alter the oxidative mechanisms of mitochondria, intact algal cells or marigold leaf tissues. The capacity for chlorophyll and carotenoid synthesis by Euglena cells was unaffected by these wavelengths. 23 references, 2 figures, 4 tables.

Study on heavy metal absorption capability of chlamidomonas reinhardtii in solution containing uranium and lead

International Nuclear Information System (INIS)

Nguyen Thuy Binh

2003-01-01

The mutant strain chlamydomonas reinhardtii No.4 obtained by C 5+ ion beam irradiation could be grown in simple mineral salt medium with initial pH range of 3.5-7.5 with continued illumination of 12,000 lux under aeration. The study demonstrated that the mutant strain C.reinhardtii had a good growth in mineral salt medium containing U 6+ (concentration about 0.015 mg/ml) and Pb 2+ (concentration about 65% and Pb 2+ about 60% from solution was estimated by analyzing dried cell. (NTB)

Metabolic flux analysis of heterotrophic growth in Chlamydomonas reinhardtii.

Directory of Open Access Journals (Sweden)

Nanette R Boyle

Full Text Available Despite the wealth of knowledge available for C. reinhardtii, the central metabolic fluxes of growth on acetate have not yet been determined. In this study, 13C-metabolic flux analysis (13C-MFA was used to determine and quantify the metabolic pathways of primary metabolism in C. reinhardtii cells grown under heterotrophic conditions with acetate as the sole carbon source. Isotopic labeling patterns of compartment specific biomass derived metabolites were used to calculate the fluxes. It was found that acetate is ligated with coenzyme A in the three subcellular compartments (cytosol, mitochondria and plastid included in the model. Two citrate synthases were found to potentially be involved in acetyl-coA metabolism; one localized in the mitochondria and the other acting outside the mitochondria. Labeling patterns demonstrate that Acetyl-coA synthesized in the plastid is directly incorporated in synthesis of fatty acids. Despite having a complete TCA cycle in the mitochondria, it was also found that a majority of the malate flux is shuttled to the cytosol and plastid where it is converted to oxaloacetate providing reducing equivalents to these compartments. When compared to predictions by flux balance analysis, fluxes measured with 13C-MFA were found to be suboptimal with respect to biomass yield; C. reinhardtii sacrifices biomass yield to produce ATP and reducing equivalents.

Photosynthetic efficiency of Chlamydomonas reinhardtii in attenuated, flashing light

NARCIS (Netherlands)

Vejrazka, C.; Janssen, M.G.J.; Streefland, M.; Wijffels, R.H.

2012-01-01

As a result of mixing and light attenuation, algae in a photobioreactor (PBR) alternate between light and dark zones and, therefore, experience variations in photon flux density (PFD). These variations in PFD are called light/dark (L/D) cycles. The objective of this study was to determine how these

Production and characterization of algae extract from Chlamydomonas reinhardtii

Directory of Open Access Journals (Sweden)

Weston Kightlinger

2014-01-01

Conclusions: This study showed that algae extract derived from C. reinhardtii is similar, if not superior, to commercially available yeast extract in nutrient content and effects on the growth and metabolism of E. coli and S. cerevisiae. Bacto™ yeast extract is valued at USD $0.15–0.35 per gram, if algae extract was sold at similar prices, it would serve as a high-value co-product in algae-based fuel processes.

ATP Production in Chlamydomonas reinhardtii Flagella by Glycolytic Enzymes

DEFF Research Database (Denmark)

Mitchell, Beth F; Pedersen, Lotte B; Feely, Michael

2005-01-01

reside in the detergent-soluble (membrane + matrix) compartments. We further show that axonemal enolase is a subunit of the CPC1 central pair complex and that reduced flagellar enolase levels in the cpc1 mutant correlate with the reduced flagellar ATP concentrations and reduced in vivo beat frequencies...

Growth performance and biochemical composition of nineteen microalgae collected from different Moroccan reservoirs

Directory of Open Access Journals (Sweden)

EL. A. IDRISSI ABDELKHALEK

2016-03-01

Full Text Available Macro- and microalgae have recently received much attention due to their valuable chemical constituents. In order to increase existing data, the authors studied nineteen microalgae species isolated from different reservoirs in the Fez region (northern Morocco, undertaking experiments to determine for each species the specific growth rate, their total amounts of proteins, carbohydrates and lipids and the influence of the growth phase on these chemical constituents. Conditions of cultivation were as follows: light intensity equal to 300 μmol photons m-2 s-1, with a temperature regime of 25/20°C (day/night and a 16/8 (light/dark photoperiod cycle. The growth rates of the nineteen studied species of microalgae showed a wide variation between species, ranging from 0.27 g l-1 d-1 for Chlamydomonas ovalis to 3.64 g l-1 d-1 for Chlorococcum wemmeri. Protein, carbohydrate and lipid contents varied greatly between taxa and within genera. Ankistrodesmus falcatus, Chlamydomonas ovalis, Chlorococcum sp., Hyaloraphidium contortum, Scenedesmus protuberans, and Synechocystis aquatilis tended to synthesize proteins, the concentrations exceeding 20% dry weight (DW. Ankistrodesmus falcatus, Ankistrodesmus sp., Chlorococcum wemmeri, Coenocystis sp., Isocystis sp., Lyngbya bergei, Oscillatoria amphibia, Polytoma papillatum, Scenedesmus protuberans, Scenedesmus sp. and Synechocystis aquatilis showed a high capacity for lipid storage, greater than 20% DW. For carbohydrate contents, only Scenedesmus protuberans and Scenedesmus quadricauda showed an excessive level compared to other scanned species with 29.21% and 24.76% DW, respectively.

Testing the time-of-flight model for flagellar length sensing.

Science.gov (United States)

Ishikawa, Hiroaki; Marshall, Wallace F

2017-11-07

Cilia and flagella are microtubule-based organelles that protrude from the surface of most cells, are important to the sensing of extracellular signals, and make a driving force for fluid flow. Maintenance of flagellar length requires an active transport process known as intraflagellar transport (IFT). Recent studies reveal that the amount of IFT injection negatively correlates with the length of flagella. These observations suggest that a length-dependent feedback regulates IFT. However, it is unknown how cells recognize the length of flagella and control IFT. Several theoretical models try to explain this feedback system. We focused on one of the models, the "time-of-flight" model, which measures the length of flagella on the basis of the travel time of IFT protein in the flagellar compartment. We tested the time-of-flight model using Chlamydomonas dynein mutant cells, which show slower retrograde transport speed. The amount of IFT injection in dynein mutant cells was higher than that in control cells. This observation does not support the prediction of the time-of-flight model and suggests that Chlamydomonas uses another length-control feedback system rather than that described by the time-of-flight model. © 2017 Ishikawa and Marshall. This article is distributed by The American Society for Cell Biology under license from the author(s). Two months after publication it is available to the public under an Attribution–Noncommercial–Share Alike 3.0 Unported Creative Commons License (http://creativecommons.org/licenses/by-nc-sa/3.0).

Monitoring Growth and Lipid Production of Some Egyptian Microalgae

International Nuclear Information System (INIS)

El-Baghdady, K.Z.; Zakaria, A.E.; Mousa, L.A.; Sadek, H.N.; Abd El Fatah, H.M.

2016-01-01

Microalgae bio diesel is a green and renewable energy resource. This study aims to examine growth and lipid production by various isolates of icroalgae using different growth media and lipid extraction techniques. Ten microalgae isolates were isolated from different samples collected from Egypt. The purified isolates were identified microscopically as: Lyngbya confervoides, Phormidium bohneri, Oscillatoria pseudogeminata, Amorphonostoc sp., Nostoc paludosum, Anabaena sphaerica related to cyanobacteria (blue green algae) and Chlorella vulgaris, Chlorella ellipsoidea, Scened esmusacutus acutus, Chlamydomonas globose related to green algae. These organisms were cultivated on two media: Bold's Basal Medium(BBM medium) and Blue Green Medium (BG-11 medium) to examine the favorite medium which supports the growth of each isolate In order to examine lipid production potentials by cyanobacterial isolates and green microalgae, two solvent systems were applied for lipid extraction, the first was (Chloroform - methanol 1:1 ) and the second was (Hexane-ethanol 1:1). Chlorella vulgaris and Anabaena sphaerica were selected as models of green microalgae and cyanobacteria espectively. Hexane-ethanol solvent system revealed higher lipid extraction capacity as compared to Chloroform- methanol system. A comparison between ten organisms for lipid production was carried out by the selected solvent mixture. The percentages of lipid to dry weight produced by Oscillatoria pseudogeminata and Chlamydomonas globose were 19.8% and14 .6% respectively recording the highest lipid to dry weight percentage. They can be considered as a promising lipid producing microalgae

Uso de una secuencia nucleotídica que regula el momento de la floración, plantas que la expresan y método para producirlas

OpenAIRE

Valverde, Federico; Serrano, Aurelio; Romero, José M.

2009-01-01

Uso de una secuencia nucleotídica que regula el momento de la floración, plantas que la expresan y método para producirlas. La presente invención se refiere a una secuencia nucleotídica que codifica para una secuencia aminoacídica de un alga verde unicelular, Chlamydomonas reinhartii, que es expresada en una célula vegetal procedente de una planta capaz de producir flores. La expresión de la secuencia se utiliza para modificar el momento de la floración de una plant...

Process and reactor design for biophotolytic hydrogen production.

Science.gov (United States)

Tamburic, Bojan; Dechatiwongse, Pongsathorn; Zemichael, Fessehaye W; Maitland, Geoffrey C; Hellgardt, Klaus

2013-07-14

The green alga Chlamydomonas reinhardtii has the ability to produce molecular hydrogen (H2), a clean and renewable fuel, through the biophotolysis of water under sulphur-deprived anaerobic conditions. The aim of this study was to advance the development of a practical and scalable biophotolytic H2 production process. Experiments were carried out using a purpose-built flat-plate photobioreactor, designed to facilitate green algal H2 production at the laboratory scale and equipped with a membrane-inlet mass spectrometry system to accurately measure H2 production rates in real time. The nutrient control method of sulphur deprivation was used to achieve spontaneous H2 production following algal growth. Sulphur dilution and sulphur feed techniques were used to extend algal lifetime in order to increase the duration of H2 production. The sulphur dilution technique proved effective at encouraging cyclic H2 production, resulting in alternating Chlamydomonas reinhardtii recovery and H2 production stages. The sulphur feed technique enabled photobioreactor operation in chemostat mode, resulting in a small improvement in H2 production duration. A conceptual design for a large-scale photobioreactor was proposed based on these experimental results. This photobioreactor has the capacity to enable continuous and economical H2 and biomass production using green algae. The success of these complementary approaches demonstrate that engineering advances can lead to improvements in the scalability and affordability of biophotolytic H2 production, giving increased confidence that H2 can fulfil its potential as a sustainable fuel of the future.

Robust expression of a bioactive mammalian protein in chlamydomonas chloroplast

Science.gov (United States)

Mayfield, Stephen P.

2010-03-16

Methods and compositions are disclosed to engineer chloroplast comprising heterologous mammalian genes via a direct replacement of chloroplast Photosystem II (PSII) reaction center protein coding regions to achieve expression of recombinant protein above 5% of total protein. When algae is used, algal expressed protein is produced predominantly as a soluble protein where the functional activity of the peptide is intact. As the host algae is edible, production of biologics in this organism for oral delivery or proteins/peptides, especially gut active proteins, without purification is disclosed.

Photolysis of caged phosphatidic acid induces flagellar excision in Chlamydomonas.

NARCIS (Netherlands)

Goedhart, J.; Gadella, Th.W.J.

2004-01-01

Phosphatidic (PtdOH) acid formation is recognized as an important step in numerous signaling pathways in both plants and mammals. To study the role of this lipid in signaling pathways, it is of major interest to be able to increase the amount of this lipid directly. Therefore, "caged" PtdOH was

Uso de una secuencia nucleotídica que regula el momento de la floración, plantas que la expresan y método para producirlas

OpenAIRE

Valverde, Federico; Serrano, Aurelio; Romero, José M.

2009-01-01

[ES] La presente invención se refiere a una secuencia nucleotídica que codifica para una secuencia aminoacídica de un alga verde unicelular, Chlamydomonas reinhartii, que es expresada en una célula vegetal procedente de una planta capaz de producir flores. La expresión de la secuencia se utiliza para modificar el momento de la floración de una planta que comprende las células vegetales transfectadas. Asimismo, la presente invención se refiere a un método para producir las células y/o las plan...

A Method for Microalgae Proteomics Analysis Based on Modified Filter-Aided Sample Preparation.

Science.gov (United States)

Li, Song; Cao, Xupeng; Wang, Yan; Zhu, Zhen; Zhang, Haowei; Xue, Song; Tian, Jing

2017-11-01

With the fast development of microalgal biofuel researches, the proteomics studies of microalgae increased quickly. A filter-aided sample preparation (FASP) method is widely used proteomics sample preparation method since 2009. Here, a method of microalgae proteomics analysis based on modified filter-aided sample preparation (mFASP) was described to meet the characteristics of microalgae cells and eliminate the error caused by over-alkylation. Using Chlamydomonas reinhardtii as the model, the prepared sample was tested by standard LC-MS/MS and compared with the previous reports. The results showed mFASP is suitable for most of occasions of microalgae proteomics studies.

Efficient recovery of uranium using genetically improved microalgae; Recuperacion eficaz de uranio utilizando microalgas geneticamente mejoradas

Energy Technology Data Exchange (ETDEWEB)

Lopez-Rodas, V.; Conde Vilda, E.; Garcia-Balboa, C.

2015-07-01

We propose an alternative process for the efficient recovery of dissolved uranium based on genetically improved microalgae. We isolate Chlamydomonas cf. fonticola from a pond extremely contaminated by uranium (∼ 25 ppm) from ENUSA U-mine, Saelices (Salamanca, Spain). After a process of genetic improvement we obtained a strain capable to recover 115 mg of U per g of dry weight, by mean of bio-adsorption on the cell wall (mostly) and intra-cytoplasm bioaccumulation. Such a genetically improved microalgae resist extremes of acidity and pollution, but even its dead biomass is still able to recover a large amount of uranium. (Author)

A new process for the fractionation of uranium; Un nuevo procedimiento para el fraccionamiento de uranio

Energy Technology Data Exchange (ETDEWEB)

Costas, E.; Baselga, B.; Tarin, F.

2015-07-01

We propose a new biological process for uranium isotopic fractionation based on Chlamydomonas cf. fonticola (microalgae) isolated from a pond extremely contaminated by uranium (. 25 ppm) from the ENUSA mine in Saelices (Salamanca, Spain) and genetically improved. The metabolic activity of this genetically improved ChlSPGI strain allows recover 115 mg of U per gram of micoralgal biomass in a short time (because this strain complete their cell cycle in . 24 hours). During this process ChlSPGI microalgae selectively captures {sup 2}35U conducting an isotopic enrichment of {sup 2}35U ({sup 2}35U δ = + 3,983%). (Author)

Dicty_cDB: Contig-U16580-1 [Dicty_cDB

Lifescience Database Archive (English)

Full Text Available 0.0 10 ( DQ015713 ) Chlamydomonas chlamydogama strain UTEX 102 26S ri... 375 0.0 9 ( EU272562 ) Eudendrium racemosum voucher persona...A gene, part... 420 0.0 9 ( EU272561 ) Ectopleura dumortieri voucher personal:A. Lindner... 396 0.0 10 ( AY0...0 9 ( EU272552 ) Clava multicornis voucher personal:P. Schuchert:S... 396 0.0 9 ( EU272549 ) Bougainvillia carolinensis voucher perso... l... 396 0.0 9 ( EU272571 ) Koellikerina fasciculata voucher personal:P. Schu...... ribosoma... 357 0.0 10 ( EU272574 ) Lizzia blondina voucher personal:P. Schuchert:Sch... 396 0.0 9 ( EU2725

Oxygen as a product of water radiolysis in high-LET tracks. II. Radiobiological implications

International Nuclear Information System (INIS)

Baverstock, K.F.; Burns, W.G.

1981-01-01

Consideration is given to the possibility that molecular oxygen generated in the tracks of energetic heavy ions is responsible for the reduction in oxygen enhancement ratio (OER) with increasing linear energy transfer (LET) observed for the loss of reproductive capacity caused by radiation in many cellular organisms. Yields of oxygen relationship of OER to LET for two organisms, Chlamydomonas reinhardii and Shigella flexneri, using a simple diffusion kinetic model for radiobiological action which takes account of the diffusion of oxygen after its formation. The results of these calculations show that the model accounts well for the shape of the OER vs. LET relationship

Effects of TiO{sub 2} nanoparticles on the growth and metabolism of three species of freshwater algae

Energy Technology Data Exchange (ETDEWEB)

Cardinale, Bradley J., E-mail: bradcard@umich.edu [University of Michigan, School of Natural Resources and Environment (United States); Bier, Raven [Duke University, Department of Biology (United States); Kwan, Courtney [Evolution and Marine Biology, University of California, Department of Ecology (United States)

2012-08-15

We examined how TiO{sub 2} nanoparticles (nTiO{sub 2}) impact the growth and metabolism of three species of freshwater green algae (Scenedesmus quadricauda, Chlamydomonas moewusii, and Chlorella vulgaris) that are widespread throughout North America. We exposed laboratory cultures to five initial concentrations of nTiO{sub 2} (0, 50, 100, 200, and 300 ppm) and measured impacts on species population growth rates, as well as on metabolic rates of gross primary production (GPP) and respiration (R). Population growth rates were consistently reduced by nTiO{sub 2}, with reduction ranging from 11 to 27 % depending on the species. But the mechanisms of reduction differed among species. For Chlamydomonas, nTiO{sub 2} reduced both GPP and R, but effects on GPP were stronger. As a consequence, carbon was respired more quickly than it was fixed, leading to reduced growth. In contrast, nTiO{sub 2} stimulated both GPP and R in Chorella. But because R was stimulated to a greater extent than GPP, carbon loss again exceeded fixation, leading to reduced growth. For Scenedesmus, nTiO{sub 2} had no significant impact on R, but reduced GPP. This pattern also caused carbon loss to exceed fixation. Results suggest that nTiO{sub 2} may generally suppress the growth of pelagic algae, but these impacts are manifest through contrasting effects on species-specific metabolic functions. Because growth and metabolism of algae are fundamental to the functioning of ecosystems and the structure of aquatic food-webs, our study suggests nTiO{sub 2} has potential to alter important community and ecosystem properties of freshwater habitats.

Effects of TiO2 nanoparticles on the growth and metabolism of three species of freshwater algae

Science.gov (United States)

Cardinale, Bradley J.; Bier, Raven; Kwan, Courtney

2012-08-01

We examined how TiO2 nanoparticles ( nTiO2) impact the growth and metabolism of three species of freshwater green algae ( Scenedesmus quadricauda, Chlamydomonas moewusii, and Chlorella vulgaris) that are widespread throughout North America. We exposed laboratory cultures to five initial concentrations of nTiO2 (0, 50, 100, 200, and 300 ppm) and measured impacts on species population growth rates, as well as on metabolic rates of gross primary production (GPP) and respiration ( R). Population growth rates were consistently reduced by nTiO2, with reduction ranging from 11 to 27 % depending on the species. But the mechanisms of reduction differed among species. For Chlamydomonas, nTiO2 reduced both GPP and R, but effects on GPP were stronger. As a consequence, carbon was respired more quickly than it was fixed, leading to reduced growth. In contrast, nTiO2 stimulated both GPP and R in Chorella. But because R was stimulated to a greater extent than GPP, carbon loss again exceeded fixation, leading to reduced growth. For Scenedesmus, nTiO2 had no significant impact on R, but reduced GPP. This pattern also caused carbon loss to exceed fixation. Results suggest that nTiO2 may generally suppress the growth of pelagic algae, but these impacts are manifest through contrasting effects on species-specific metabolic functions. Because growth and metabolism of algae are fundamental to the functioning of ecosystems and the structure of aquatic food-webs, our study suggests nTiO2 has potential to alter important community and ecosystem properties of freshwater habitats.

Effects of TiO2 nanoparticles on the growth and metabolism of three species of freshwater algae

International Nuclear Information System (INIS)

Cardinale, Bradley J.; Bier, Raven; Kwan, Courtney

2012-01-01

We examined how TiO 2 nanoparticles (nTiO 2 ) impact the growth and metabolism of three species of freshwater green algae (Scenedesmus quadricauda, Chlamydomonas moewusii, and Chlorella vulgaris) that are widespread throughout North America. We exposed laboratory cultures to five initial concentrations of nTiO 2 (0, 50, 100, 200, and 300 ppm) and measured impacts on species population growth rates, as well as on metabolic rates of gross primary production (GPP) and respiration (R). Population growth rates were consistently reduced by nTiO 2 , with reduction ranging from 11 to 27 % depending on the species. But the mechanisms of reduction differed among species. For Chlamydomonas, nTiO 2 reduced both GPP and R, but effects on GPP were stronger. As a consequence, carbon was respired more quickly than it was fixed, leading to reduced growth. In contrast, nTiO 2 stimulated both GPP and R in Chorella. But because R was stimulated to a greater extent than GPP, carbon loss again exceeded fixation, leading to reduced growth. For Scenedesmus, nTiO 2 had no significant impact on R, but reduced GPP. This pattern also caused carbon loss to exceed fixation. Results suggest that nTiO 2 may generally suppress the growth of pelagic algae, but these impacts are manifest through contrasting effects on species-specific metabolic functions. Because growth and metabolism of algae are fundamental to the functioning of ecosystems and the structure of aquatic food-webs, our study suggests nTiO 2 has potential to alter important community and ecosystem properties of freshwater habitats.

Development of a Biosensor for Environmental Monitoring Based on Microalgae Immobilized in Silica Hydrogels

Directory of Open Access Journals (Sweden)

Claude Durrieu

2012-12-01

Full Text Available A new biosensor was designed for the assessment of aquatic environment quality. Three microalgae were used as toxicity bioindicators: Chlorella vulgaris, Pseudokirchneriella subcapitata and Chlamydomonas reinhardtii. These microalgae were immobilized in alginate and silica hydrogels in a two step procedure. After studying the growth rate of entrapped cells, chlorophyll fluorescence was measured after exposure to (3-(3,4-dichlorophenyl-1,1-dimethylurea (DCMU and various concentrations of the common herbicide atrazine. Microalgae are very sensitive to herbicides and detection of fluorescence enhancement with very good efficiency was realized. The best detection limit was 0.1 µM, obtained with the strain C. reinhardtii after 40 minutes of exposure.

Effect of petroleum hydrocarbons on algae

International Nuclear Information System (INIS)

Bhadauria, S.; Sengar, R.M.S.; Mittal, S.; Bhattacharjee, S.

1992-01-01

Algal species (65) were isolated from oil refinery effluent. Twenty-five of these species were cultured in Benecke's medium in a growth chamber, along with controls. Retardation in algal growth, inhibition in algal photosynthesis, and discoloration was observed in petroleum enriched medium. Few forms, viz. Cyclotella sp., Cosmarium sp., and Merismopedia sp. could not survive. The lag phase lengthened by several days and slope of exponential phase was also depressed. Chlamydomonas sp., Scenedesmus sp., Ankistrodesmus sp., Nitzschia sp. and Navicula sp. were comparatively susceptible to petroleum. Depression in carbon fixation, cell numbers, and total dry algal mass was noticeable, showing toxicity to both diatoms and green algae

AcEST: BP915429 [AcEST

Lifescience Database Archive (English)

Full Text Available 0XF0|Q00XF0_OSTTA Homology to unknown gene (Fragment) OS=Os... 126 5e-28 tr|Q4VGM4|Q4VGM4_CHLRE Low photochemical bleaching... 1 protein OS=... 125 8e-28 tr|Q4VGM5|Q4VGM5_CHLRE Low photochemical bleaching...38 P++ Sbjct: 376 PVM 378 >tr|Q4VGM4|Q4VGM4_CHLRE Low photochemical bleaching 1 protein OS=Chlamydomonas rei...ATHR 8 LPG+I NL K + Y D LG R Sbjct: 615 LLPGLIFNLNKKVSYTDPLGGPSR 638 >tr|Q4VGM5|Q4VGM5_CHLRE Low photochemical bleaching

Highly conserved small subunit residues influence rubisco large subunit catalysis.

Science.gov (United States)

Genkov, Todor; Spreitzer, Robert J

2009-10-30

The chloroplast enzyme ribulose 1,5-bisphosphate carboxylase/oxygenase (Rubisco) catalyzes the rate-limiting step of photosynthetic CO(2) fixation. With a deeper understanding of its structure-function relationships and competitive inhibition by O(2), it may be possible to engineer an increase in agricultural productivity and renewable energy. The chloroplast-encoded large subunits form the active site, but the nuclear-encoded small subunits can also influence catalytic efficiency and CO(2)/O(2) specificity. To further define the role of the small subunit in Rubisco function, the 10 most conserved residues in all small subunits were substituted with alanine by transformation of a Chlamydomonas reinhardtii mutant that lacks the small subunit gene family. All the mutant strains were able to grow photosynthetically, indicating that none of the residues is essential for function. Three of the substitutions have little or no effect (S16A, P19A, and E92A), one primarily affects holoenzyme stability (L18A), and the remainder affect catalysis with or without some level of associated structural instability (Y32A, E43A, W73A, L78A, P79A, and F81A). Y32A and E43A cause decreases in CO(2)/O(2) specificity. Based on the x-ray crystal structure of Chlamydomonas Rubisco, all but one (Glu-92) of the conserved residues are in contact with large subunits and cluster near the amino- or carboxyl-terminal ends of large subunit alpha-helix 8, which is a structural element of the alpha/beta-barrel active site. Small subunit residues Glu-43 and Trp-73 identify a possible structural connection between active site alpha-helix 8 and the highly variable small subunit loop between beta-strands A and B, which can also influence Rubisco CO(2)/O(2) specificity.

Chronic uranium exposure and growth toxicity for phytoplankton. Dose-effect relationship: first comparison of chemical and radiological toxicity

International Nuclear Information System (INIS)

Gilbin, R.; Pradines, C.; Garnier-Laplace, J.

2004-01-01

The bioavailability of uranium for freshwater organisms, as for other dissolved metals, is closely linked to chemical speciation in solution (U aqueous speciation undergoes tremendous changes in the presence of ligands commonly found in natural waters e.g. carbonate, phosphate, hydroxide and natural organic matter). For the studied chemical domain, short-term uranium uptake experiments have already shown that the free uranyl ion concentration [UO 2 2+ ] is a good predictor of uranium uptake by the green algae Chlamydomonas reinhardtii, as predicted by the Free Ion Activity Model. In agreement with these results, acidic pH and low ligands concentrations in water enhance uranium bioavailability and consequently its potential chronic effects on phytoplankton. Moreover, uranium is known to be both radio-toxic and chemo-toxic. The use of different isotopes of uranium allows to expose organisms to different radiological doses for the same molar concentration: e.g. for a given element concentration (chemical dose), replacing depleted U by U-233 obviously leads to an enhanced radiological delivered dose to organisms (x10 4 ). In this work we established relationships between uranium doses (depleted uranium and 233-U ) and effect on the growth rate of the green algae Chlamydomonas reinhardtii. Uranium bioaccumulation was also monitored. Growth rate was measured both in classical batch (0-72 hrs) and continuous (turbidostat) cultures, the latter protocol allowing medium renewal to diminish exudates accumulation and speciation changes in the medium. The differences in effects will be, if possible, related to the development of defence mechanisms against the formation of reactive oxygen species (forms of glutathione) and the production of phyto-chelatins (small peptides rich in cystein that play an important role in the homeostasis and the detoxication of metals in cells). (author)

EGFR tyrosine kinase inhibitory peptide attenuates Helicobacter pylori-mediated hyper-proliferation in AGS enteric epithelial cells

Energy Technology Data Exchange (ETDEWEB)

Himaya, S.W.A. [Marine Bio-Process Research Center, Pukyong National University, Nam-Gu, Busan, 608-737 (Korea, Republic of); Dewapriya, Pradeep [Department of Chemistry, Pukyong National University, Nam-Gu, Busan, 608-737 (Korea, Republic of); Kim, Se-Kwon, E-mail: sknkim@pknu.ac.kr [Marine Bio-Process Research Center, Pukyong National University, Nam-Gu, Busan, 608-737 (Korea, Republic of); Department of Chemistry, Pukyong National University, Nam-Gu, Busan, 608-737 (Korea, Republic of)

2013-06-15

Helicobacter pylori infection is one of the most critical causes of stomach cancer. The current study was conducted to explore the protective effects of an isolated active peptide H-P-6 (Pro-Gln-Pro-Lys-Val-Leu-Asp-Ser) from microbial hydrolysates of Chlamydomonas sp. against H. pylori-induced carcinogenesis. The peptide H-P-6 has effectively suppressed H. pylori-induced hyper-proliferation and migration of gastric epithelial cells (AGS). However, the peptide did not inhibit the viability of the bacteria or invasion into AGS cells. Therefore, the effect of the peptide on regulating H. pylori-induced molecular signaling was investigated. The results indicated that H. pylori activates the EGFR tyrosine kinase signaling and nuclear translocation of the β-catenin. The EGFR activation has led to the up-regulation of PI3K/Akt signaling pathway. Moreover, the nuclear translocation levels of β-catenin were significantly increased as a result of Akt mediated down-regulation of GSK3/β protein levels in the cytoplasm. Both of these consequences have resulted in increased expression of cell survival and migration related genes such as c-Myc, cyclin-D, MMP-2 and matrilysin. Interestingly, the isolated peptide potently inhibited H. pylori-mediated EGFR activation and thereby down-regulated the subsequent P13K/Akt signaling leading to β-catenin nuclear translocation. The effect of the peptide was confirmed with the use of EGFR tyrosine kinase inhibitor AG1487 and molecular docking studies. Collectively this study identifies a potent peptide which regulates the H. pylori-induced hyper-proliferation and migration of AGS cells at molecular level. - Highlights: • Chlamydomonas sp. derived peptide H-P-6 inhibits H. pylori-induced pathogenesis. • H-P-6 suppresses H. pylori-induced hyper-proliferation and migration of AGS cells. • The peptide inhibits H. pylori-induced EGFR activation.

Proton transfers in a channelrhodopsin-1 studied by Fourier transform infrared (FTIR) difference spectroscopy and site-directed mutagenesis.

Science.gov (United States)

Ogren, John I; Yi, Adrian; Mamaev, Sergey; Li, Hai; Spudich, John L; Rothschild, Kenneth J

2015-05-15

Channelrhodopsin-1 from the alga Chlamydomonas augustae (CaChR1) is a low-efficiency light-activated cation channel that exhibits properties useful for optogenetic applications such as a slow light inactivation and a red-shifted visible absorption maximum as compared with the more extensively studied channelrhodopsin-2 from Chlamydomonas reinhardtii (CrChR2). Previously, both resonance Raman and low-temperature FTIR difference spectroscopy revealed that unlike CrChR2, CaChR1 under our conditions exhibits an almost pure all-trans retinal composition in the unphotolyzed ground state and undergoes an all-trans to 13-cis isomerization during the primary phototransition typical of other microbial rhodopsins such as bacteriorhodopsin (BR). Here, we apply static and rapid-scan FTIR difference spectroscopy along with site-directed mutagenesis to characterize the proton transfer events occurring upon the formation of the long-lived conducting P2 (380) state of CaChR1. Assignment of carboxylic C=O stretch bands indicates that Asp-299 (homolog to Asp-212 in BR) becomes protonated and Asp-169 (homolog to Asp-85 in BR) undergoes a net change in hydrogen bonding relative to the unphotolyzed ground state of CaChR1. These data along with earlier FTIR measurements on the CaChR1 → P1 transition are consistent with a two-step proton relay mechanism that transfers a proton from Glu-169 to Asp-299 during the primary phototransition and from the Schiff base to Glu-169 during P2 (380) formation. The unusual charge neutrality of both Schiff base counterions in the P2 (380) conducting state suggests that these residues may function as part of a cation selective filter in the open channel state of CaChR1 as well as other low-efficiency ChRs. © 2015 by The American Society for Biochemistry and Molecular Biology, Inc.

Investigation of the chemical identity of soluble organophosphorus compounds found in natural waters. Research report

International Nuclear Information System (INIS)

Minear, R.A.

1978-04-01

Four algal species (Chlamydomonas reinhardtii, Chlorella pyrenoidosa, Anacystis nidulans, and Anabaena flos-aquae) were grown in batch culture on 32 P labelled media to yield dissolved organic phosphorus (DOP) compounds containing a radioactive tag. The DOP compounds of filtered culture solutions were characterized by Sephadex gel filtration and thin layer chromatography (TLC) as a function of culture age. Additional TLC of individual Sephadex fractions was conducted. Time, culture and known compounds (inositol mono- and hexaphosphate) comparisons were made. High performance liquid chromatography was used to separate inositol mono- and hexaphosphates and to compare the DOP components of one algal species (C. reinhardtii) with inositol phosphates. Combinations of alkaline bromination and Sephadex pretreatment were examined

Properties of the intracellular transient receptor potential (TRP) channel in yeast, Yvc1.

Science.gov (United States)

Chang, Yiming; Schlenstedt, Gabriel; Flockerzi, Veit; Beck, Andreas

2010-05-17

Transient receptor potential (TRP) channels are found among mammals, flies, worms, ciliates, Chlamydomonas, and yeast but are absent in plants. These channels are believed to be tetramers of proteins containing six transmembrane domains (TMs). Their primary structures are diverse with sequence similarities only in some short amino acid sequence motifs mainly within sequences covering TM5, TM6, and adjacent domains. In the yeast genome, there is one gene encoding a TRP-like sequence. This protein forms an ion channel in the vacuolar membrane and is therefore called Yvc1 for yeast vacuolar conductance 1. In the following we summarize its prominent features. Copyright 2009 Federation of European Biochemical Societies. Published by Elsevier B.V. All rights reserved.

Heterologous expression of an algal hydrogenase in a heterocystous cyanobacterium

Energy Technology Data Exchange (ETDEWEB)

Thorsten Heidorn; Peter Lindblad [Dept. of Physiological Botany, Uppsala University, Villavogen 6, SE-752 36 Uppsala, (Sweden)

2006-07-01

For the expression of an active algal [FeFe] hydrogenase in the heterocystous cyanobacterium Nostoc punctiforme A TCC 29133 the Chlamydomonas reinhardtii hydrogenase gene hydA1 and the accessory genes hydEF and hydG are to be introduced into the cyanobacteria cells. The genes were amplified by PCR from EST clones, cloned into the cloning vector pBluescript SK+ and sequenced. An expression vector for multi-cistronic cloning, based on pSCR202, was constructed and for a functional test GFP was inserted as a reporter gene. The GFP construct was transformed into Nostoc punctiforme A TCC 29133 by electroporation and expression of GFP was visualized by fluorescence microscopy. (authors)

Heterologous expression of an algal hydrogenase in a hetero-cystous cyanobacterium

Energy Technology Data Exchange (ETDEWEB)

Thorsten Heidorn; Peter Lindblad [Dept. of Physiological Botany, Uppsala University, V illavagen 6, SE-752 36 Uppsala, (Sweden)

2006-07-01

For the expression of an active algal [FeFe] hydrogenase in the hetero-cystous cyanobacterium Nostoc punctiforme A TCC 29133 the Chlamydomonas reinhardtii hydrogenase gene hydA1 and the accessory genes hydEF and hydG are to be introduced into the cyano-bacterial cells. The genes were amplified by PCR from EST clones, cloned into the cloning vector pBluescript SK+ and sequenced. An expression vector for multi-cistronic cloning, based on pSCR202, was constructed and for a functional test GFP was inserted as a reporter gene. The GFP construct was transformed into Nostoc punctiforme A TCC 29133 by electroporation and expression of GFP was visualized by fluorescence microscopy. (authors)

Heterologous expression of an algal hydrogenase in a heterocystous cyanobacterium

International Nuclear Information System (INIS)

Thorsten Heidorn; Peter Lindblad

2006-01-01

For the expression of an active algal [FeFe] hydrogenase in the heterocystous cyanobacterium Nostoc punctiforme A TCC 29133 the Chlamydomonas reinhardtii hydrogenase gene hydA1 and the accessory genes hydEF and hydG are to be introduced into the cyanobacteria cells. The genes were amplified by PCR from EST clones, cloned into the cloning vector pBluescript SK+ and sequenced. An expression vector for multi-cistronic cloning, based on pSCR202, was constructed and for a functional test GFP was inserted as a reporter gene. The GFP construct was transformed into Nostoc punctiforme A TCC 29133 by electroporation and expression of GFP was visualized by fluorescence microscopy. (authors)

Heterologous expression of an algal hydrogenase in a hetero-cystous cyanobacterium

International Nuclear Information System (INIS)

Thorsten Heidorn; Peter Lindblad

2006-01-01

For the expression of an active algal [FeFe] hydrogenase in the hetero-cystous cyanobacterium Nostoc punctiforme A TCC 29133 the Chlamydomonas reinhardtii hydrogenase gene hydA1 and the accessory genes hydEF and hydG are to be introduced into the cyano-bacterial cells. The genes were amplified by PCR from EST clones, cloned into the cloning vector pBluescript SK+ and sequenced. An expression vector for multi-cistronic cloning, based on pSCR202, was constructed and for a functional test GFP was inserted as a reporter gene. The GFP construct was transformed into Nostoc punctiforme A TCC 29133 by electroporation and expression of GFP was visualized by fluorescence microscopy. (authors)

Effects of γ-rays on the survival of several unicellular green algae and the relation to saprobity

International Nuclear Information System (INIS)

Hamada, Jin; Saito, Masahiro; Bando, Tadashi; Ishida, M.R.

1990-01-01

The lethal effects of 60 Co-γ-rays on the cells of Chlamydomonas reinhardi Dangeard (Chlorophyceae) and four species of desmids (Charophyceae) such as Closterium acerosum Ehrenberg, Netrium digitus Ehrenberg, Closterium ehrenbergii Meneghini, and Pleurotaenium ehrenbergii de Bary were studied at their plateau phase. Wide variation in their response to γ-rays was observed. Among these algae. Cl.acerosum, which is the most tolerant against saprobity, showed the highest resistance to γ-rays. The values of D 37 for Cl.acerosum, N. digitus, Cl.ehrenbergii, Ch.reihardi and Pl.ehrenbergii were about 210Gy, 155Gy, 60Gy and 30Gy, respectively. Some relationship between the resistance to radiation and tolerance to saprobity existed among closely related species. (author)

Effects of LET on production and repair of damage in Chlamydomonas reinhardi

International Nuclear Information System (INIS)

Braby, L.A.; Jacobson, B.S.

1983-01-01

Modeling activity was directed toward a more complete understanding of the kinetics of the processes affecting the reproductive survival of these cells. The need for additional types of data to test aspects of this model is evident

Carbon and nitrogen stable isotope ratios analysis of food sources for Chironomus acerbiphilus larvae (Diptera Chironomidae) in strongly acidic lake Katanuma

International Nuclear Information System (INIS)

Doi, Hideyuki; Kikuchi, Eisuke; Shikano, Shuichi

2001-01-01

The food sources for Chironomus acerbiphilus larvae (Diptera Chironomidae) were analyzed using carbon and nitrogen stable isotope ratios in Lake Katanuma. Lake Katanuma is a volcanic and strongly acidic lake (average pH 2.2). In Lake Katanuma, potential sources of diets for the chironomid larvae are limited including a benthic diatom (Pinnularia braunii), a phytoplankton (Chlamydomonas acidophila), sulfate oxidizing bacteria, and vascular plants supplied from vegetation surrounding the lake. Based on the average carbon and nitrogen isotope ratios among these potential sources and sediment, benthic diatoms were considered to be most probable food source of the chironomid larvae. δ 13 C values of the chironomid were significantly different among seasons and habitat depths, suggesting that diet of C. acerbiphilus changed seasonally and with habitat depth. (author)

TOR (target of rapamycin) is a key regulator of triacylglycerol accumulation in microalgae.

Science.gov (United States)

Imamura, Sousuke; Kawase, Yasuko; Kobayashi, Ikki; Shimojima, Mie; Ohta, Hiroyuki; Tanaka, Kan

2016-01-01

Most microalgae abundantly accumulate lipid droplets (LDs) containing triacylglycerols (TAGs) under several stress conditions, but the underlying molecular mechanism of this accumulation remains unclear. In a recent study, we found that inhibition of TOR (target of rapamycin), a highly conserved protein kinase of eukaryotes, by rapamycin resulted in TAG accumulation in microalgae, indicating that TOR negatively regulates TAG accumulation. Here, we show that formation of intracellular LDs and TAG accumulation were also induced in the unicellular green alga Chlamydomonas reinhardtii after exposure to Torin1 or AZD8055, which are novel TOR inhibitors that inhibit TOR activity in a manner different from rapamycin. These results supported quite well our previous conclusion that TOR is a central regulator of TAG accumulation in microalgae.

The oxygen in the track hypothesis: microdosimetric implications

International Nuclear Information System (INIS)

Baverstock, K.F.; Burns, W.G.; May, R.

1978-01-01

We have applied diffusion kinetics to a simple competition between self-repair, self-fixation and reaction with oxygen to model the LET dependence of the OER in two cellular organisms, Shigella flexneri and Chlamydomonas reinhardii, using track yields of oxygen suggested by the experiments with metal ions, and obtain good agreement with the data of Alper and Bryant (1974). The calculations suggest that the reaction with track oxygen is almost complete before that with added oxygen begins, and they provide an order-of-magnitude estimate for the G value of initial lesions. We conclude that the dependence of OER on LET is not directly linked to microdosimetric considerations and that the processes which determine RBE take place before intra-track reactions

A magnetic trap for living cells suspended in a paramagnetic buffer

Science.gov (United States)

Winkleman, Adam; Gudiksen, Katherine L.; Ryan, Declan; Whitesides, George M.; Greenfield, Derek; Prentiss, Mara

2004-09-01

This manuscript describes the fabrication and use of a three-dimensional magnetic trap for diamagnetic objects in an aqueous solution of paramagnetic ions; this trap uses permanent magnets. It demonstrates trapping of polystyrene spheres, and of various types of living cells: mouse fibroblast (NIH-3T3), yeast (Saccharomyces cerevisiae), and algae (Chlamydomonas reinhardtii). For a 40mM solution of gadolinium (III) diethylenetriaminepentaacetic acid (Gd .DTPA) in aqueous buffer, the smallest cell (particle) that could be trapped had a radius of ˜2.5μm. The trapped particle and location of the magnetic trap can be translated in three dimensions by independent manipulation of the permanent magnets. This letter a1so characterizes the biocompatibility of the trapping solution.

Directed evolution and in silico analysis of reaction centre proteins reveal molecular signatures of photosynthesis adaptation to radiation pressure.

Directory of Open Access Journals (Sweden)

Giuseppina Rea

2011-01-01

Full Text Available Evolutionary mechanisms adopted by the photosynthetic apparatus to modifications in the Earth's atmosphere on a geological time-scale remain a focus of intense research. The photosynthetic machinery has had to cope with continuously changing environmental conditions and particularly with the complex ionizing radiation emitted by solar flares. The photosynthetic D1 protein, being the site of electron tunneling-mediated charge separation and solar energy transduction, is a hot spot for the generation of radiation-induced radical injuries. We explored the possibility to produce D1 variants tolerant to ionizing radiation in Chlamydomonas reinhardtii and clarified the effect of radiation-induced oxidative damage on the photosynthetic proteins evolution. In vitro directed evolution strategies targeted at the D1 protein were adopted to create libraries of chlamydomonas random mutants, subsequently selected by exposures to radical-generating proton or neutron sources. The common trend observed in the D1 aminoacidic substitutions was the replacement of less polar by more polar amino acids. The applied selection pressure forced replacement of residues more sensitive to oxidative damage with less sensitive ones, suggesting that ionizing radiation may have been one of the driving forces in the evolution of the eukaryotic photosynthetic apparatus. A set of the identified aminoacidic substitutions, close to the secondary plastoquinone binding niche and oxygen evolving complex, were introduced by site-directed mutagenesis in un-transformed strains, and their sensitivity to free radicals attack analyzed. Mutants displayed reduced electron transport efficiency in physiological conditions, and increased photosynthetic performance stability and oxygen evolution capacity in stressful high-light conditions. Finally, comparative in silico analyses of D1 aminoacidic sequences of organisms differently located in the evolution chain, revealed a higher ratio of residues

Mutation of Growth Arrest Specific 8 Reveals a Role in Motile Cilia Function and Human Disease.

Science.gov (United States)

Lewis, Wesley R; Malarkey, Erik B; Tritschler, Douglas; Bower, Raqual; Pasek, Raymond C; Porath, Jonathan D; Birket, Susan E; Saunier, Sophie; Antignac, Corinne; Knowles, Michael R; Leigh, Margaret W; Zariwala, Maimoona A; Challa, Anil K; Kesterson, Robert A; Rowe, Steven M; Drummond, Iain A; Parant, John M; Hildebrandt, Friedhelm; Porter, Mary E; Yoder, Bradley K; Berbari, Nicolas F

2016-07-01

Ciliopathies are genetic disorders arising from dysfunction of microtubule-based cellular appendages called cilia. Different cilia types possess distinct stereotypic microtubule doublet arrangements with non-motile or 'primary' cilia having a 9+0 and motile cilia have a 9+2 array of microtubule doublets. Primary cilia are critical sensory and signaling centers needed for normal mammalian development. Defects in their structure/function result in a spectrum of clinical and developmental pathologies including abnormal neural tube and limb patterning. Altered patterning phenotypes in the limb and neural tube are due to perturbations in the hedgehog (Hh) signaling pathway. Motile cilia are important in fluid movement and defects in motility result in chronic respiratory infections, altered left-right asymmetry, and infertility. These features are the hallmarks of Primary Ciliary Dyskinesia (PCD, OMIM 244400). While mutations in several genes are associated with PCD in patients and animal models, the genetic lesion in many cases is unknown. We assessed the in vivo functions of Growth Arrest Specific 8 (GAS8). GAS8 shares strong sequence similarity with the Chlamydomonas Nexin-Dynein Regulatory Complex (NDRC) protein 4 (DRC4) where it is needed for proper flagella motility. In mammalian cells, the GAS8 protein localizes not only to the microtubule axoneme of motile cilia, but also to the base of non-motile cilia. Gas8 was recently implicated in the Hh signaling pathway as a regulator of Smoothened trafficking into the cilium. Here, we generate the first mouse with a Gas8 mutation and show that it causes severe PCD phenotypes; however, there were no overt Hh pathway phenotypes. In addition, we identified two human patients with missense variants in Gas8. Rescue experiments in Chlamydomonas revealed a subtle defect in swim velocity compared to controls. Further experiments using CRISPR/Cas9 homology driven repair (HDR) to generate one of these human missense variants in

Mutation of Growth Arrest Specific 8 Reveals a Role in Motile Cilia Function and Human Disease.

Directory of Open Access Journals (Sweden)

Wesley R Lewis

2016-07-01

Full Text Available Ciliopathies are genetic disorders arising from dysfunction of microtubule-based cellular appendages called cilia. Different cilia types possess distinct stereotypic microtubule doublet arrangements with non-motile or 'primary' cilia having a 9+0 and motile cilia have a 9+2 array of microtubule doublets. Primary cilia are critical sensory and signaling centers needed for normal mammalian development. Defects in their structure/function result in a spectrum of clinical and developmental pathologies including abnormal neural tube and limb patterning. Altered patterning phenotypes in the limb and neural tube are due to perturbations in the hedgehog (Hh signaling pathway. Motile cilia are important in fluid movement and defects in motility result in chronic respiratory infections, altered left-right asymmetry, and infertility. These features are the hallmarks of Primary Ciliary Dyskinesia (PCD, OMIM 244400. While mutations in several genes are associated with PCD in patients and animal models, the genetic lesion in many cases is unknown. We assessed the in vivo functions of Growth Arrest Specific 8 (GAS8. GAS8 shares strong sequence similarity with the Chlamydomonas Nexin-Dynein Regulatory Complex (NDRC protein 4 (DRC4 where it is needed for proper flagella motility. In mammalian cells, the GAS8 protein localizes not only to the microtubule axoneme of motile cilia, but also to the base of non-motile cilia. Gas8 was recently implicated in the Hh signaling pathway as a regulator of Smoothened trafficking into the cilium. Here, we generate the first mouse with a Gas8 mutation and show that it causes severe PCD phenotypes; however, there were no overt Hh pathway phenotypes. In addition, we identified two human patients with missense variants in Gas8. Rescue experiments in Chlamydomonas revealed a subtle defect in swim velocity compared to controls. Further experiments using CRISPR/Cas9 homology driven repair (HDR to generate one of these human missense

Rapid synthesis of gold and silver nanoparticles using tryptone as a reducing and capping agent

Science.gov (United States)

Mehta, Sourabh M.; Sequeira, Marilyn P.; Muthurajana, Harries; D'Souza, Jacinta S.

2018-02-01

Due to its eco-friendliness, recent times have seen an immense interest in the green synthesis of metallic nanoparticles. We present here, a protocol for the rapid and cheap synthesis of Au and Ag nanoparticles (NPs) using 1 mg/ml tryptone (trypsinized casein) as a reducing and capping agent. These nanoparticles are spherical, 10 nm in diameter and relatively monodispersed. The atoms of these NPs are arranged in face-centered cubic fashion. Further, when tested for their cytotoxic property against HeLa and VERO cell lines, gold nanoparticles were more lethal than silver nanoparticles, with a more or less similar trend observed against both Gram-positive and Gram-negative bacteria. On the other hand, the NPs were least cytotoxic against a unicellular alga, Chlamydomonas reinhardtii implying their eco-friendly property.

Modelization of tritium transfer into the organic compartments of algae

International Nuclear Information System (INIS)

Bonotto, S.; Gerber, G.B.; Arapis, G.; Kirchmann, R.

1982-01-01

Uptake of tritium oxide and its conversion into organic tritium was studied in four different types of algae with widely varying size and growth characteristics (Acetabularia acetabulum, Boergesenia forbesii, two strains of Chlamydomonas and Dunaliella bioculata). Water in the cell and the vacuales equilibrates rapidly with external tritium water. Tritium is actively incorporated into organically bound form as the organisms grow. During the stationary phase, incorporation of tritium is slow. There exists a discrimination against the incorporation of tritium into organically bound form. A model has been elaborated taking in account these different factors. It appears that transfer of organic tritium by algae growing near the sites of release would be significant only for actively growing algae. Algae growing slowly may, however, be useful as cumulative indicators of discontinuous tritium release. (author)

Carbon and nitrogen stable isotope ratios analysis of food sources for Chironomus acerbiphilus larvae (Diptera Chironomidae) in strongly acidic lake Katanuma

Energy Technology Data Exchange (ETDEWEB)

Doi, Hideyuki [Tohoku Univ., Sendai (Japan). Graduate School of Science; Kikuchi, Eisuke; Shikano, Shuichi

2001-12-01

The food sources for Chironomus acerbiphilus larvae (Diptera Chironomidae) were analyzed using carbon and nitrogen stable isotope ratios in Lake Katanuma. Lake Katanuma is a volcanic and strongly acidic lake (average pH 2.2). In Lake Katanuma, potential sources of diets for the chironomid larvae are limited including a benthic diatom (Pinnularia braunii), a phytoplankton (Chlamydomonas acidophila), sulfate oxidizing bacteria, and vascular plants supplied from vegetation surrounding the lake. Based on the average carbon and nitrogen isotope ratios among these potential sources and sediment, benthic diatoms were considered to be most probable food source of the chironomid larvae. {delta}{sup 13}C values of the chironomid were significantly different among seasons and habitat depths, suggesting that diet of C. acerbiphilus changed seasonally and with habitat depth. (author)

Growth response of four freshwater algal species to dissolved organic nitrogen of different concentration and complexity

DEFF Research Database (Denmark)

Fiedler, Dorothea; Graeber, Daniel; Badrian, Maria

2015-01-01

1. Dissolved organic nitrogen (DON) compounds dominate the nitrogen pool of many lakes, but their importance as nitrogen sources for freshwater phytoplankton is not fully understood. Previous growth experiments demonstrated the availability of urea and amino acids but often at unnaturally high...... (DCAA), natural organic matter (NOM)) or with nitrate as the sole nitrogen source. Monocultures of Chlamydomonas spp., Cyclotella meneghiniana, Microcystis aeruginosa and Anabaena flos-aquae were incubated with dissolved nitrogen compounds at concentrations ranging from 0.01 to 0.5 mg N L−1, which...... and their compound preferences. Therefore, DON composition can influence biomass and structure of phytoplankton communities. 6. These experiments demonstrate the importance of the main DON compounds for phytoplankton growth when no inorganic nitrogen is available. DON should in future be included in nitrogen budget...

Phycoremediation as a potential water decontamination method

International Nuclear Information System (INIS)

Tatarova, D.; Galanda, D.; Kuruc, J.

2017-01-01

In experiments, we focused on the determination of the phycoremediation potential of Chlamydomonas reinhardtii and Scenedesmus obliquus in targeted contaminated aqueous solutions containing radioisotopes 137 Cs and 6 0Co. Microalgae were selected based on their high bioremediation capability. Phycoremediation potential was determined by monitoring the effect of different pH values between pH 2 to pH 9 as well as by monitoring the decrease in activity of the solution over time. Cultivation of microalgae took place in 12 h/12 h light/dark light mode in blue and red light, which promotes plant growth at room temperature. In order to determine the micro-sorption capacity, a method was used to determine the concentration of microns using a Buerker cell in parallel with the spectrophotometric method. (authors)

Structure/Function of the Novel Proteins LCIB and LCIC in the Chlamydomonas CCM

Energy Technology Data Exchange (ETDEWEB)

Martin, Spalding H. [Iowa State Univ., Ames, IA (United States). Dept. of Genetics, Development and Cell Biology

2017-05-09

The goal of this project was to investigate the function of two novel proteins, LCIB and LCIC, which together form an essential protein complex that is required for function of a carbon-dioxide-concentrating mechanism (CCM) required by microalgae to grow in environments where carbon dioxide levels are at or below air equilibration levels.

Factors Affecting the Mating Competence in the Unicellular Green Alga Chlamydomonas eugametos (Volvocales)

Czech Academy of Sciences Publication Activity Database

Zachleder, Vilém; Hendrychová, Jana; Bišová, Kateřina; Kubín, Štěpán

2002-01-01

Roč. 47, č. 1 (2002), s. 69-72 ISSN 0015-5632 R&D Projects: GA ČR GA204/97/0576; GA AV ČR IAC5020012 Institutional research plan: CEZ:AV0Z5020903 Keywords : routineli * prepared * gametes Subject RIV: EB - Genetics ; Molecular Biology Impact factor: 0.979, year: 2002

Photosynthetic efficiency and oxygen evolution of Chlamydomonas reinhardtii under continuous and flashing light.

NARCIS (Netherlands)

Vejrazka, C.; Janssen, M.; Benvenuti, G.; Streefland, M.; Wijffels, R.H.

2013-01-01

As a result of mixing and light attenuation in a photobioreactor (PBR), microalgae experience light/dark (L/D) cycles that can enhance PBR efficiency. One parameter which characterizes L/D cycles is the duty cycle; it determines the time fraction algae spend in the light. The objective of this study

How to track protists in three dimensions

Science.gov (United States)

Drescher, Knut; Leptos, Kyriacos C.; Goldstein, Raymond E.

2009-01-01

We present an apparatus optimized for tracking swimming micro-organisms in the size range of 10-1000 μm, in three dimensions (3Ds), far from surfaces, and with negligible background convective fluid motion. Charge coupled device cameras attached to two long working distance microscopes synchronously image the sample from two perpendicular directions, with narrow band dark-field or bright-field illumination chosen to avoid triggering a phototactic response. The images from the two cameras can be combined to yield 3D tracks of the organism. Using additional, highly directional broad-spectrum illumination with millisecond timing control the phototactic trajectories in 3D of organisms ranging from Chlamydomonas to Volvox can be studied in detail. Surface-mediated hydrodynamic interactions can also be investigated without convective interference. Minimal modifications to the apparatus allow for studies of chemotaxis and other taxes.

Photoinduced conductivity in mycosporine-like amino acids

International Nuclear Information System (INIS)

Kulkarni, Atul; Lee, Jeong Hun; Seo, Hyo Hyun; Kim, Hyoung-Shik; Cho, Moon Jin; Shin, Dong Sun; Kim, Taesung; Moh, Sang Hyun

2015-01-01

Mycosporine-like amino acids (MAAs) are an important group of novel bioactive compounds having immense biotechnological potentials due to their UV screening properties and antioxidant activities. However, their photoelectric properties were not evaluated yet. In the present work two types of MAAs Shinorine and Porphyra-334, were extracted from algae; Chlamydomonas hedlyei and Porphyra yezoensis respectively and its electrical transport properties were investigated upon illumination of UV light. The combination of optical absorption and electron transport measurement of MAAs in a field effect transistor device reveals that these changes are mainly due to the carboxyl group present in MAAs. This study reports a platform technology for the development of novel biochemical–electrical devices. - Highlights: • MAAs FET shows photoelectric effect upon UV illumination. • Enhancement in photo conductance is due to the hydroxyl ethyl group. • Potential as bio-opto-electrical devices applications

Cell disruption and lipid extraction for microalgal biorefineries: A review.

Science.gov (United States)

Lee, Soo Youn; Cho, Jun Muk; Chang, Yong Keun; Oh, You-Kwan

2017-11-01

The microalgae-based biorefinement process has attracted much attention from academic and industrial researchers attracted to its biofuel, food and nutraceutical applications. In this paper, recent developments in cell-disruption and lipid-extraction methods, focusing on four biotechnologically important microalgal species (namely, Chlamydomonas, Haematococcus, Chlorella, and Nannochloropsis spp.), are reviewed. The structural diversity and rigidity of microalgal cell walls complicate the development of efficient downstream processing methods for cell-disruption and subsequent recovery of intracellular lipid and pigment components. Various mechanical, chemical and biological cell-disruption methods are discussed in detail and compared based on microalgal species and status (wet/dried), scale, energy consumption, efficiency, solvent extraction, and synergistic combinations. The challenges and prospects of the downstream processes for the future development of eco-friendly and economical microalgal biorefineries also are outlined herein. Copyright © 2017 Elsevier Ltd. All rights reserved.

Evidence for a photoprotective function for secondary carotenoids of snow algae

International Nuclear Information System (INIS)

Bidigare, R.R.; Ondrusek, M.E.; Kennicutt, M.C. II; Iturriaga, R.; Harvey, H.R.; Hoham, R.W.; Macko, S.A.

1993-01-01

Snow algae occupy a unique habitat in high altitude and polar environments. These algae are often subject to extremes in nutrient availability, acidity, solar irradiance, desiccation, and ambient temperature. This report documents the accumulation of secondary carotenoids by snow algae in response to the availability of nitrogenous nutrients. Unusually large accumulations of astaxanthin esters in extra-chloroplastic lipid globules produce the characteristic red pigmentation typical of some snow algae (e.g., Chlamydomonas nivalis (Bauer) Wille). Consequently, these compounds greatly reduce the amount of light available for absorption by the light-harvesting pigment-protein complexes, thus potentially limiting photoinhibition and photodamage caused by intense solar radiation. The esterification of astaxanthin with fatty acids represents a possible mechanism by which this chromophore can be concentrated within cytoplasmic globules to maximize its photoprotective efficiency. 53 refs., 2 figs., 4 tabs

Three-dimensional structural analysis of eukaryotic flagella/cilia by electron cryo-tomography

International Nuclear Information System (INIS)

Bui, Khanh Huy; Pigino, Gaia; Ishikawa, Takashi

2011-01-01

Based on the molecular architecture revealed by electron cryo-tomography, the mechanism of the bending motion of eukaryotic flagella/cilia is discussed. Electron cryo-tomography is a potential approach to analyzing the three-dimensional conformation of frozen hydrated biological macromolecules using electron microscopy. Since projections of each individual object illuminated from different orientations are merged, electron tomography is capable of structural analysis of such heterogeneous environments as in vivo or with polymorphism, although radiation damage and the missing wedge are severe problems. Here, recent results on the structure of eukaryotic flagella, which is an ATP-driven bending organelle, from green algae Chlamydomonas are presented. Tomographic analysis reveals asymmetric molecular arrangements, especially that of the dynein motor proteins, in flagella, giving insight into the mechanism of planar asymmetric bending motion. Methodological challenges to obtaining higher-resolution structures from this technique are also discussed

Irreducible Representations of Oscillatory and Swirling Flows in Active Soft Matter

Science.gov (United States)

Ghose, Somdeb; Adhikari, R.

2014-03-01

Recent experiments imaging fluid flow around swimming microorganisms have revealed complex time-dependent velocity fields that differ qualitatively from the stresslet flow commonly employed in theoretical descriptions of active matter. Here we obtain the most general flow around a finite sized active particle by expanding the surface stress in irreducible Cartesian tensors. This expansion, whose first term is the stresslet, must include, respectively, third-rank polar and axial tensors to minimally capture crucial features of the active oscillatory flow around translating Chlamydomonas and the active swirling flow around rotating Volvox. The representation provides explicit expressions for the irreducible symmetric, antisymmetric, and isotropic parts of the continuum active stress. Antisymmetric active stresses do not conserve orbital angular momentum and our work thus shows that spin angular momentum is necessary to restore angular momentum conservation in continuum hydrodynamic descriptions of active soft matter.

The AtCAO gene, encoding chlorophyll a oxygenase, is required for chlorophyll b synthesis in Arabidopsis thaliana

Science.gov (United States)

Espineda, Cromwell E.; Linford, Alicia S.; Devine, Domenica; Brusslan, Judy A.

1999-01-01

Chlorophyll b is synthesized from chlorophyll a and is found in the light-harvesting complexes of prochlorophytes, green algae, and both nonvascular and vascular plants. We have used conserved motifs from the chlorophyll a oxygenase (CAO) gene from Chlamydomonas reinhardtii to isolate a homologue from Arabidopsis thaliana. This gene, AtCAO, is mutated in both leaky and null chlorina1 alleles, and DNA sequence changes cosegregate with the mutant phenotype. AtCAO mRNA levels are higher in three different mutants that have reduced levels of chlorophyll b, suggesting that plants that do not have sufficient chlorophyll b up-regulate AtCAO gene expression. Additionally, AtCAO mRNA levels decrease in plants that are grown under dim-light conditions. We have also found that the six major Lhcb proteins do not accumulate in the null ch1-3 allele. PMID:10468639

Photoinduced conductivity in mycosporine-like amino acids

Energy Technology Data Exchange (ETDEWEB)

Kulkarni, Atul; Lee, Jeong Hun; Seo, Hyo Hyun; Kim, Hyoung-Shik; Cho, Moon Jin; Shin, Dong Sun [Antiaging Research Institute of BIO-FD and C Co. Ltd., Incheon 406-840 (Korea, Republic of); Kim, Taesung [Sungkyunkwan Advanced Institute of Nanotechnology (SAINT), Sungkyunkwan University, Suwon 440-746 (Korea, Republic of); Moh, Sang Hyun, E-mail: shmoh@biofdnc.com [Antiaging Research Institute of BIO-FD and C Co. Ltd., Incheon 406-840 (Korea, Republic of)

2015-02-01

Mycosporine-like amino acids (MAAs) are an important group of novel bioactive compounds having immense biotechnological potentials due to their UV screening properties and antioxidant activities. However, their photoelectric properties were not evaluated yet. In the present work two types of MAAs Shinorine and Porphyra-334, were extracted from algae; Chlamydomonas hedlyei and Porphyra yezoensis respectively and its electrical transport properties were investigated upon illumination of UV light. The combination of optical absorption and electron transport measurement of MAAs in a field effect transistor device reveals that these changes are mainly due to the carboxyl group present in MAAs. This study reports a platform technology for the development of novel biochemical–electrical devices. - Highlights: • MAAs FET shows photoelectric effect upon UV illumination. • Enhancement in photo conductance is due to the hydroxyl ethyl group. • Potential as bio-opto-electrical devices applications.

OMNY—A tOMography Nano crYo stage

Science.gov (United States)

Holler, M.; Raabe, J.; Diaz, A.; Guizar-Sicairos, M.; Wepf, R.; Odstrcil, M.; Shaik, F. R.; Panneels, V.; Menzel, A.; Sarafimov, B.; Maag, S.; Wang, X.; Thominet, V.; Walther, H.; Lachat, T.; Vitins, M.; Bunk, O.

2018-04-01

For many scientific questions gaining three-dimensional insight into a specimen can provide valuable information. We here present an instrument called "tOMography Nano crYo (OMNY)," dedicated to high resolution 3D scanning x-ray microscopy at cryogenic conditions via hard X-ray ptychography. Ptychography is a lens-less imaging method requiring accurate sample positioning. In OMNY, this in achieved via dedicated laser interferometry and closed-loop position control reaching sub-10 nm positioning accuracy. Cryogenic sample conditions are maintained via conductive cooling. 90 K can be reached when using liquid nitrogen as coolant, and 10 K is possible with liquid helium. A cryogenic sample-change mechanism permits measurements of cryogenically fixed specimens. We compare images obtained with OMNY with older measurements performed using a nitrogen gas cryo-jet of stained, epoxy-embedded retina tissue and of frozen-hydrated Chlamydomonas cells.

Hydrogen production by photosynthetic water splitting

Energy Technology Data Exchange (ETDEWEB)

Greenbaum, E.; Lee, J.W.; Tevault, C.V. [Oak Ridge National Lab., TN (United States)

1996-10-01

Contrary to the prediction of the {open_quotes}Z-scheme{close_quotes} model of photosynthesis, experiments demonstrated that mutants of Chlamydomonas containing Photosystem II (PSII) but lacking Photosystem I (PSI), can grow photoautotrophically with O{sub 2} evolution and using atmospheric CO{sub 2} as the sole carbon source. Autotrophic photosynthesis by PSI-deficient mutants was stable both under anaerobic conditions and in air (21% O{sub 2}) at an actinic intensity of 200 {mu}E/m{sup -2}{sup {sm_bullet}}s. This {open_quotes}PSII photosynthesis,{close_quotes} sufficient to support cell development and mobility, may also occur in wild-type green algae and higher plants. The mutants can survive under 2000 {mu}E{sup {sm_bullet}}m{sup -2}{sup {sm_bullet}}s{sup -1} with air, although they have less resistance to photoinhibition.

Copper economy in Chlamydomonas: Prioritized allocation and reallocation of copper to respiration vs. photosynthesis

Science.gov (United States)

Kropat, Janette; Gallaher, Sean D.; Urzica, Eugen I.; Nakamoto, Stacie S.; Strenkert, Daniela; Tottey, Stephen; Mason, Andrew Z.; Merchant, Sabeeha S.

2015-01-01

Inorganic elements, although required only in trace amounts, permit life and primary productivity because of their functions in catalysis. Every organism has a minimal requirement of each metal based on the intracellular abundance of proteins that use inorganic cofactors, but elemental sparing mechanisms can reduce this quota. A well-studied copper-sparing mechanism that operates in microalgae faced with copper deficiency is the replacement of the abundant copper protein plastocyanin with a heme-containing substitute, cytochrome (Cyt) c6. This switch, which is dependent on a copper-sensing transcription factor, copper response regulator 1 (CRR1), dramatically reduces the copper quota. We show here that in a situation of marginal copper availability, copper is preferentially allocated from plastocyanin, whose function is dispensable, to other more critical copper-dependent enzymes like Cyt oxidase and a ferroxidase. In the absence of an extracellular source, copper allocation to Cyt oxidase includes CRR1-dependent proteolysis of plastocyanin and quantitative recycling of the copper cofactor from plastocyanin to Cyt oxidase. Transcriptome profiling identifies a gene encoding a Zn-metalloprotease, as a candidate effecting copper recycling. One reason for the retention of genes encoding both plastocyanin and Cyt c6 in algal and cyanobacterial genomes might be because plastocyanin provides a competitive advantage in copper-depleted environments as a ready source of copper. PMID:25646490

Synthesis of chlorophyll b: Localization of chlorophyllide a oxygenase and discovery of a stable radical in the catalytic subunit

Science.gov (United States)

Eggink, Laura L; LoBrutto, Russell; Brune, Daniel C; Brusslan, Judy; Yamasato, Akihiro; Tanaka, Ayumi; Hoober, J Kenneth

2004-01-01

Background Assembly of stable light-harvesting complexes (LHCs) in the chloroplast of green algae and plants requires synthesis of chlorophyll (Chl) b, a reaction that involves oxygenation of the 7-methyl group of Chl a to a formyl group. This reaction uses molecular oxygen and is catalyzed by chlorophyllide a oxygenase (CAO). The amino acid sequence of CAO predicts mononuclear iron and Rieske iron-sulfur centers in the protein. The mechanism of synthesis of Chl b and localization of this reaction in the chloroplast are essential steps toward understanding LHC assembly. Results Fluorescence of a CAO-GFP fusion protein, transiently expressed in young pea leaves, was found at the periphery of mature chloroplasts and on thylakoid membranes by confocal fluorescence microscopy. However, when membranes from partially degreened cells of Chlamydomonas reinhardtii cw15 were resolved on sucrose gradients, full-length CAO was detected by immunoblot analysis only on the chloroplast envelope inner membrane. The electron paramagnetic resonance spectrum of CAO included a resonance at g = 4.3, assigned to the predicted mononuclear iron center. Instead of a spectrum of the predicted Rieske iron-sulfur center, a nearly symmetrical, approximately 100 Gauss peak-to-trough signal was observed at g = 2.057, with a sensitivity to temperature characteristic of an iron-sulfur center. A remarkably stable radical in the protein was revealed by an isotropic, 9 Gauss peak-to-trough signal at g = 2.0042. Fragmentation of the protein after incorporation of 125I- identified a conserved tyrosine residue (Tyr-422 in Chlamydomonas and Tyr-518 in Arabidopsis) as the radical species. The radical was quenched by chlorophyll a, an indication that it may be involved in the enzymatic reaction. Conclusion CAO was found on the chloroplast envelope and thylakoid membranes in mature chloroplasts but only on the envelope inner membrane in dark-grown C. reinhardtii cells. Such localization provides further

Desempenho de sistema de tratamento de lixiviado de aterro sanitário com recirculação do efluente Performance of a sanitary landfill leachate treatment system with effluent recirculation

Directory of Open Access Journals (Sweden)

Cláudia Lavina Martins

2010-12-01

Full Text Available O objetivo deste estudo foi avaliar o desempenho de um sistema de tratamento de lixiviado de aterro sanitário, em escala piloto, formado por três lagoas em série (L1, L2 e L3 seguidas por um filtro de pedras (FP. Foram estudadas três condições operacionais, verificando-se o efeito da recirculação do efluente da L3 para a L1: 0, 50 e 100% da vazão. O sistema absorveu bem as flutuações de cargas, apresentando remoções superiores a 80% para DBO F, 70% para DQO T e 98% para nitrogênio amoniacal. Na lagoa L2 houve nitrificação parcial, com acúmulo de nitritos. Ocorreu presença marcante do gênero Chlamydomonas nas lagoas L2 e L3 fotossintéticas. Observou-se diminuição da toxicidade do lixiviado, com reduções superiores a 95% na saída do sistema. A melhor eficiência de remoção para as três condições operacionais estudadas foi obtida com 100% de recirculação.The objective of this study was to evaluate the performance of a landfill leachate treatment system, at pilot scale, formed by three in series ponds (L1, L2, and L3 followed by a rock filter. Three operational conditions were studied, assessing the effluent recirculation effect from L3 to L1: 0, 50 and 100% of the flow rate. The system assimilated well the load fluctuations, showing removal efficiencies over 80% for BOD F, 70% for COD T and 98% for ammonia. In the L2 pond there was partial nitrification with nitrite accumulation. It was observed an outstanding presence of the Chlamydomonas gender in photosynthetic ponds L2 and L3. A reduction of the toxicity from the landfill leachate was observed, with reduction of over 95% in the system output. The better removal efficiency for the three studied operational conditions was obtained for 100% of recirculation.

Antibiotic Algae by Chemical Surface Engineering.

Science.gov (United States)

Kerschgens, Isabel P; Gademann, Karl

2018-03-02

Chemical cell-surface engineering is a tool for modifying and altering cellular functions. Herein, we report the introduction of an antibiotic phenotype to the green alga Chlamydomonas reinhardtii by chemically modifying its cell surface. Flow cytometry and confocal microscopy studies demonstrated that a hybrid of the antibiotic vancomycin and a 4-hydroxyproline oligomer binds reversibly to the cell wall without affecting the viability or motility of the cells. The modified cells were used to inhibit bacterial growth of Gram-positive Bacillus subtilis cultures. Delivery of the antibiotic from the microalgae to the bacterial cells was verified by microscopy. Our studies provide compelling evidence that 1) chemical surface engineering constitutes a useful tool for the introduction of new, previously unknown functionality, and 2) living microalgae can serve as new platforms for drug delivery. © 2018 Wiley-VCH Verlag GmbH & Co. KGaA, Weinheim.

Modeling and cellular studies

International Nuclear Information System (INIS)

Anon.

1982-01-01

Testing the applicability of mathematical models with carefully designed experiments is a powerful tool in the investigations of the effects of ionizing radiation on cells. The modeling and cellular studies complement each other, for modeling provides guidance for designing critical experiments which must provide definitive results, while the experiments themselves provide new input to the model. Based on previous experimental results the model for the accumulation of damage in Chlamydomonas reinhardi has been extended to include various multiple two-event combinations. Split dose survival experiments have shown that models tested to date predict most but not all the observed behavior. Stationary-phase mammalian cells, required for tests of other aspects of the model, have been shown to be at different points in the cell cycle depending on how they were forced to stop proliferating. These cultures also demonstrate different capacities for repair of sublethal radiation damage

Green Algae as Model Organisms for Biological Fluid Dynamics

Science.gov (United States)

Goldstein, Raymond E.

2015-01-01

In the past decade, the volvocine green algae, spanning from the unicellular Chlamydomonas to multicellular Volvox, have emerged as model organisms for a number of problems in biological fluid dynamics. These include flagellar propulsion, nutrient uptake by swimming organisms, hydrodynamic interactions mediated by walls, collective dynamics and transport within suspensions of microswimmers, the mechanism of phototaxis, and the stochastic dynamics of flagellar synchronization. Green algae are well suited to the study of such problems because of their range of sizes (from 10 μm to several millimeters), their geometric regularity, the ease with which they can be cultured, and the availability of many mutants that allow for connections between molecular details and organism-level behavior. This review summarizes these recent developments and highlights promising future directions in the study of biological fluid dynamics, especially in the context of evolutionary biology, that can take advantage of these remarkable organisms.

Cloning and Characterization of the -Carotene Desaturase Gene from Chlorella protothecoides CS-41

Directory of Open Access Journals (Sweden)

Meiya Li

2011-01-01

Full Text Available To elucidate the lutein biosynthesis pathway in the lutein-producing alga, Chlorella protothecoides CS-41, the -carotene desaturase gene (zds was isolated from Chlorella protothecoides using the approach of rapid amplification of cDNA ends. The full-length cDNA sequence was 2031 bp and contained 1755 bp putative open reading frame which encodes a 584 amino acid deduced polypeptide whose computed molecular weight was 63.7 kDa. Sequence homology research indicated that the nucleotide and putative protein had sequence identities of 72.5% and 69.5% with those of the green alga Chlamydomonas reinhardtii, respectively. Phylogenetic analysis demonstrated that the ZDS from C. protothecoides CS-41 had a closer relationship with those of chlorophyta and higher plants than with those of other species. In addition, we also found that the zds gene expression was upregulated in response to light.

Universal entrainment mechanism controls contact times with motile cells

Science.gov (United States)

Mathijssen, Arnold J. T. M.; Jeanneret, Raphaël; Polin, Marco

2018-03-01

Contact between particles and motile cells underpins a wide variety of biological processes, from nutrient capture and ligand binding to grazing, viral infection, and cell-cell communication. The window of opportunity for these interactions depends on the basic mechanism determining contact time, which is currently unknown. By combining experiments on three different species—Chlamydomonas reinhardtii, Tetraselmis subcordiforms, and Oxyrrhis marina—with simulations and analytical modeling, we show that the fundamental physical process regulating proximity to a swimming microorganism is hydrodynamic particle entrainment. The resulting distribution of contact times is derived within the framework of Taylor dispersion as a competition between advection by the cell surface and microparticle diffusion, and predicts the existence of an optimal tracer size that is also observed experimentally. Spatial organization of flagella, swimming speed, and swimmer and tracer size influence entrainment features and provide tradeoffs that may be tuned to optimize the estimated probabilities for microbial interactions like predation and infection.

Carbon and hydrogen metabolism of green algae in light and dark

Energy Technology Data Exchange (ETDEWEB)

1990-01-01

After adaptation to a hydrogen metabolism, Chlamydomonas reinhardtii can photoanaerobically metabolize acetate with the evolution of H{sub 2} and CO{sub 2}. An enzyme profile of the chloroplastic, cytoplasmic, and mitochondrial fractions were obtained with a cellular fractionation procedure that incorporated cell wall removal by autolysine, digestion of the plasmalemma with digitonin and fractionation by differential centrifugation on a Percoll step gradient. The sequence of events leading to the photo-evolution of H{sub 2} from acetate includes the conversion of acetate into succinate via the extraplastidic glyoxylate cycle, the oxidation of succinate to fumarate by chloroplastic succinic dehydrogenase and the oxidation of malate to oxaloacetate in the chloroplast by NAD dependent malate dehydrogenase. The level of potential activity of the enzymes was sufficient to accommodate the observed rate of gas evolution. The isolated darkened chloroplast evolves aerobically CO{sub 2} from glucose indicating a chloroplastic respiratory pathway. Evolution of CO{sub 2} is blocked by mitochondrial inhibitors.

Comparative assessment of single and joint effects of diuron and Irgarol 1051 on Arctic and temperate microalgae using chlorophyll a fluorescence imaging

KAUST Repository

Kottuparambil, Sreejith

2017-02-06

Ship groundings and ice-breakers can cause pollution of the polar environment with antifouling biocides such as diuron and Irgarol 1051. The present study used pulse amplitude modulated fluorometry to compare single and joint toxicities of diuron and Irgarol 1051 on two freshwater taxa of microalgae (Chlorella and Chlamydomonas) originating from Arctic and temperate regions. 30min acute toxicity tests using chlorophyll a (Chl a) fluorescence revealed that Arctic strains of microalgae were more sensitive to herbicides than their temperate counterparts. Diuron and Irgarol 1051 had equal toxicities in the Arctic species, while Irgarol 1051 was more toxic (EC50=5.55–14.70μgL−1) than diuron (EC50=12.90–>40μgL−1) in the temperate species. Toxicity assessment of various mixtures of diuron and Irgarol 1051 revealed antagonistic, additive, and synergistic effects. Our data suggest that herbicides can adversely affect photosynthesis in Arctic microalgae at relatively low levels, and their impact can increase under complex mixture conditions.

Salicylhydroxamic acid (SHAM) inhibition of the DIC-pump in unicellular algae

International Nuclear Information System (INIS)

Goyal, A.; Tolbert, N.E.

1989-01-01

SHAM at 1 or 2 mM inhibits dissolved inorganic carbon (DIC) concentrating mechanisms in unicellular green algae as measured by photosynthetic oxygen evolution or by 14 C-inorganic carbon uptake (using silicone oil centrifugation techniques). This inhibition was reversed by high levels of DIC whereby the cells do not require the concentrating mechanism. SHAM inhibited the DIC-pump, which uses external CO 2 , in three species of algae, Dunaliella tertiolecta, Chlamydomonas reinhardtii, and Scenedesmus obliquus when adapted to low CO 2 and assayed around neutral pH. Scenedesmus adapted to air at pH 9.0 to use external HCO 3 - were not affected by SHAM. It is important to establish low optimum concentrations of SHAM, which varied with the algal species. The mechanism of SHAM inhibition of the CO 2 concentrating process is unknown. SHAM inhibits alternative respiration in these algae, but SHAM may also inhibit other reactions involving H + gradients or transporters associated with the DIC-pump

Investigation of animal and algal bioassays for reliable saxitoxin ecotoxicity and cytotoxicity risk evaluation.

Science.gov (United States)

Perreault, François; Matias, Marcelo Seleme; Melegari, Silvia Pedroso; Pinto, Catia Regina Silva de Carvalho; Creppy, Edmond Ekué; Popovic, Radovan; Matias, William Gerson

2011-05-01

Contamination of water bodies by saxitoxin can result in various toxic effects in aquatic organisms. Saxitoxin contamination has also been shown to be a threat to human health in several reported cases, even resulting in death. In this study, we evaluated the sensitivity of animal (Neuro-2A) and algal (Chlamydomonas reinhardtii) bioassays to saxitoxin effect. Neuro-2A cells were found to be sensitive to saxitoxin, as shown by a 24 h EC50 value of 1.5 nM, which was obtained using a cell viability assay. Conversely, no saxitoxin effect was found in any of the algal biomarkers evaluated, for the concentration range tested (2-128 nM). These results indicate that saxitoxin may induce toxic effects in animal and human populations at concentrations where phytoplankton communities are not affected. Therefore, when evaluating STX risk of toxicity, algal bioassays do not appear to be reliable indicators and should always be conducted in combination with animal bioassays. Copyright © 2011 Elsevier Inc. All rights reserved.

Reaction Coordinate Leading to H2 Production in [FeFe]-Hydrogenase Identified by Nuclear Resonance Vibrational Spectroscopy and Density Functional Theory.

Science.gov (United States)

Pelmenschikov, Vladimir; Birrell, James A; Pham, Cindy C; Mishra, Nakul; Wang, Hongxin; Sommer, Constanze; Reijerse, Edward; Richers, Casseday P; Tamasaku, Kenji; Yoda, Yoshitaka; Rauchfuss, Thomas B; Lubitz, Wolfgang; Cramer, Stephen P

2017-11-22

[FeFe]-hydrogenases are metalloenzymes that reversibly reduce protons to molecular hydrogen at exceptionally high rates. We have characterized the catalytically competent hydride state (H hyd ) in the [FeFe]-hydrogenases from both Chlamydomonas reinhardtii and Desulfovibrio desulfuricans using 57 Fe nuclear resonance vibrational spectroscopy (NRVS) and density functional theory (DFT). H/D exchange identified two Fe-H bending modes originating from the binuclear iron cofactor. DFT calculations show that these spectral features result from an iron-bound terminal hydride, and the Fe-H vibrational frequencies being highly dependent on interactions between the amine base of the catalytic cofactor with both hydride and the conserved cysteine terminating the proton transfer chain to the active site. The results indicate that H hyd is the catalytic state one step prior to H 2 formation. The observed vibrational spectrum, therefore, provides mechanistic insight into the reaction coordinate for H 2 bond formation by [FeFe]-hydrogenases.

[Fe]-hydrogenases in green algae: photo-fermentation and hydrogen evolution under sulfur deprivation

Energy Technology Data Exchange (ETDEWEB)

Winkler, M.; Hemschemeier, A.; Happe, T. [Botanisches Institut der Universitat Bonn (Germany); Gotor, C. [CSIC y Universidad de Sevilla (Spain). Instituto de Bioquimica Vegetal y Fotosintesis; Melis, A. [University of California, Berkeley, CA (United States). Department of Plant and Microbial Biology

2002-12-01

Recent studies indicate that [Fe]-hydrogenases and H{sub 2} metabolism are widely distributed among green algae. The enzymes are simple structured and catalyze H{sub 2} evolution with similar rates than the more complex [Fe]-hydrogenases from bacteria. Different green algal species developed diverse strategies to survive under sulfur deprivation. Chlamydomonas reinhardtii evolves large quantities of hydrogen gas in the absence of sulfur. In a sealed culture of C. reinhardtii, the photosynthetic O{sub 2} evolution rate drops below the rate of respiratory O{sub 2} consumption due to a reversible inhibition of photosystem II, thus leading to an intracellular anaerobiosis. The algal cells survive under these anaerobic conditions by switching their metabolism to a kind of photo-fermentation. Although possessing a functional [Fe]-hydrogenase gene, the cells of Scenedesmus obliquus produce no significant amounts of H{sub 2} under S-depleted conditions. Biochemical analyses indicate that S. obliquus decreases almost the complete metabolic activities while maintaining a low level of respiratory activity. (author)

Intraflagellar transport particle size scales inversely with flagellar length: revisiting the balance-point length control model.

Science.gov (United States)

Engel, Benjamin D; Ludington, William B; Marshall, Wallace F

2009-10-05

The assembly and maintenance of eukaryotic flagella are regulated by intraflagellar transport (IFT), the bidirectional traffic of IFT particles (recently renamed IFT trains) within the flagellum. We previously proposed the balance-point length control model, which predicted that the frequency of train transport should decrease as a function of flagellar length, thus modulating the length-dependent flagellar assembly rate. However, this model was challenged by the differential interference contrast microscopy observation that IFT frequency is length independent. Using total internal reflection fluorescence microscopy to quantify protein traffic during the regeneration of Chlamydomonas reinhardtii flagella, we determined that anterograde IFT trains in short flagella are composed of more kinesin-associated protein and IFT27 proteins than trains in long flagella. This length-dependent remodeling of train size is consistent with the kinetics of flagellar regeneration and supports a revised balance-point model of flagellar length control in which the size of anterograde IFT trains tunes the rate of flagellar assembly.

Biochemical compositions and fatty acid profiles in four species of microalgae cultivated on household sewage and agro-industrial residues.

Science.gov (United States)

Calixto, Clediana Dantas; da Silva Santana, Jordana Kaline; de Lira, Evandro Bernardo; Sassi, Patrícia Giulianna Petraglia; Rosenhaim, Raul; da Costa Sassi, Cristiane Francisca; da Conceição, Marta Maria; Sassi, Roberto

2016-12-01

The potential of four regional microalgae species was evaluated in relation to their cell growth and biomass production when cultured in the following alternative media: bio-composts of fruit/horticultural wastes (HB), sugarcane waste and vinasse (VB) chicken excrements (BCE), raw chicken manure (RCM), and municipal domestic sewage (MDS). The cultures were maintained under controlled conditions and their growth responses, productivities, biochemical compositions, and the ester profiles of their biomasses were compared to the results obtained in the synthetic media. The MDS and HB media demonstrated promising results for cultivation, especially of Chlorella sp., Chlamydomonas sp., and Lagerheimia longiseta, which demonstrated productivities superior to those seen when grown on the control media. The highest lipid levels were obtained with the HB medium. The data obtained demonstrated the viability of cultivating microalgae and producing biomass in alternative media prepared from MDS and HB effluents to produce biodiesel. Copyright © 2016 Elsevier Ltd. All rights reserved.

Method for delivery of small molecules and proteins across the cell wall of algae using molecular transporters

Science.gov (United States)

Geihe, Erika; Trantow, Brian; Wender, Paul; Hyman, Joel M.; Parvin, Bahram

2017-11-14

The introduction of tools to study, control or expand the inner-workings of algae has been slow to develop. Provided are embodiments of a molecular method based on guanidinium-rich molecular transporters (GR-MoTrs) for bringing molecular cargos into algal cells. The methods of the disclosure have been shown to work in wild-type algae that have an intact cell wall. Developed using Chlamydomonas reinhardtii, this method is also successful with less studied algae, including Neochloris oleoabundans and Scenedesmus dimorphus, thus providing a new and versatile tool for algal research and modification. The method of delivering a cargo compound to an algal cell comprises contacting an algal cell with a guanidinium-rich delivery vehicle comprising a guanidinium-rich molecular transporter (GR-MoTr) linked to a cargo compound desired to be delivered to the algal cell, whereby the guanidinium-rich molecular transporter can traverse the algal cell wall, thereby delivering the cargo compound to the algal cell.

Stationary Size Distributions of Growing Cells with Binary and Multiple Cell Division

Science.gov (United States)

Rading, M. M.; Engel, T. A.; Lipowsky, R.; Valleriani, A.

2011-10-01

Populations of unicellular organisms that grow under constant environmental conditions are considered theoretically. The size distribution of these cells is calculated analytically, both for the usual process of binary division, in which one mother cell produces always two daughter cells, and for the more complex process of multiple division, in which one mother cell can produce 2 n daughter cells with n=1,2,3,… . The latter mode of division is inspired by the unicellular algae Chlamydomonas reinhardtii. The uniform response of the whole population to different environmental conditions is encoded in the individual rates of growth and division of the cells. The analytical treatment of the problem is based on size-dependent rules for cell growth and stochastic transition processes for cell division. The comparison between binary and multiple division shows that these different division processes lead to qualitatively different results for the size distribution and the population growth rates.

Chloroplast DNA codon use: evidence for selection at the psb A locus based on tRNA availability.

Science.gov (United States)

Morton, B R

1993-09-01

Codon use in the three sequenced chloroplast genomes (Marchantia, Oryza, and Nicotiana) is examined. The chloroplast has a bias in that codons NNA and NNT are favored over synonymous NNC and NNG codons. This appears to be a consequence of an overall high A + T content of the genome. This pattern of codon use is not followed by the psb A gene of all three genomes and other psb A sequences examined. In this gene, the codon use favors NNC over NNT for twofold degenerate amino acids. In each case the only tRNA coded by the genome is complementary to the NNC codon. This codon use is similar to the codon use by chloroplast genes examined from Chlamydomonas reinhardtii. Since psb A is the major translation product of the chloroplast, this suggests that selection is acting on the codon use of this gene to adapt codons to tRNA availability, as previously suggested for unicellular organisms.

Spatial and temporal variation of phytoplankton in a tropical eutrophic river.

Science.gov (United States)

Santana, L M; Moraes, M E B; Silva, D M L; Ferragut, C

2016-04-19

This study aims to evaluate the environmental factors determining of the changes in phytoplankton structure in spatial (upper, middle and lower course) and seasonal (dry and rainy period) scales in a eutrophic river (Almada River, northeastern Brazil). In the study period, total accumulated rainfall was below of the historic average, resulting in flow reduction, mainly in rainy period. High orthophosphate concentration was found at the sampling sites. Phytoplankton chlorophyll a increased from upstream to downstream. Geitlerinema splendidum (S1) and Chlamydomonas sp. (X2) were the most abundant species in the upper course and several species of diatoms (D), Euglenophyceae (W1, W2) and Chlorophyceae (X1) in the middle and lower course. The functional groups were found to be characteristic of lotic ecosystem, shallow, with low light availability, rich in organic matter and eutrophic environments. We conclude that phytoplankton community structure was sensitive to change of the river flow and nutrient availability in spatial and seasonal scale in a tropical river.

Spatial and temporal variation of phytoplankton in a tropical eutrophic river

Directory of Open Access Journals (Sweden)

L. M. Santana

Full Text Available Abstract This study aims to evaluate the environmental factors determining of the changes in phytoplankton structure in spatial (upper, middle and lower course and seasonal (dry and rainy period scales in a eutrophic river (Almada River, northeastern Brazil. In the study period, total accumulated rainfall was below of the historic average, resulting in flow reduction, mainly in rainy period. High orthophosphate concentration was found at the sampling sites. Phytoplankton chlorophyll a increased from upstream to downstream. Geitlerinema splendidum (S1 and Chlamydomonas sp. (X2 were the most abundant species in the upper course and several species of diatoms (D, Euglenophyceae (W1, W2 and Chlorophyceae (X1 in the middle and lower course. The functional groups were found to be characteristic of lotic ecosystem, shallow, with low light availability, rich in organic matter and eutrophic environments. We conclude that phytoplankton community structure was sensitive to change of the river flow and nutrient availability in spatial and seasonal scale in a tropical river.

A Rapid and Simple Bioassay Method for Herbicide Detection

Directory of Open Access Journals (Sweden)

Xiu-Qing Li

2008-01-01

Full Text Available Chlamydomonas reinhardtii, a unicellular green alga, has been used in bioassay detection of a variety of toxic compounds such as pesticides and toxic metals, but mainly using liquid culture systems. In this study, an algal lawn--agar system for semi-quantitative bioassay of herbicidal activities has been developed. Sixteen different herbicides belonging to 11 different categories were applied to paper disks and placed on green alga lawns in Petri dishes. Presence of herbicide activities was indicated by clearing zones around the paper disks on the lawn 2-3 days after application. The different groups of herbicides induced clearing zones of variable size that depended on the amount, mode of action, and chemical properties of the herbicides applied to the paper disks. This simple, paper-disk-algal system may be used to detect the presence of herbicides in water samples and act as a quick and inexpensive semi-quantitative screening for assessing herbicide contamination.

CCDC103 mutations cause primary ciliary dyskinesia by disrupting assembly of ciliary dynein arms

Science.gov (United States)

Panizzi, Jennifer R.; Becker-Heck, Anita; Castleman, Victoria H.; Al-Mutairi, Dalal; Liu, Yan; Loges, Niki T.; Pathak, Narendra; Austin-Tse, Christina; Sheridan, Eamonn; Schmidts, Miriam; Olbrich, Heike; Werner, Claudius; Häffner, Karsten; Hellman, Nathan; Chodhari, Rahul; Gupta, Amar; Kramer-Zucker, Albrecht; Olale, Felix; Burdine, Rebecca D.; Schier, Alexander F.; O’Callaghan, Christopher; Chung, Eddie MK; Reinhardt, Richard; Mitchison, Hannah M.; King, Stephen M.; Omran, Heymut; Drummond, Iain A.

2012-01-01

Cilia are essential for fertilization, respiratory clearance, cerebrospinal fluid circulation, and to establish laterality1. Cilia motility defects cause Primary Ciliary Dyskinesia (PCD, MIM 242650), a disorder affecting 1:15-30,000 births. Cilia motility requires the assembly of multisubunit dynein arms that drive cilia bending2. Despite progress in understanding the genetic basis of PCD, mutations remain to be identified for several PCD linked loci3. Here we show that the zebrafish cilia paralysis mutant schmalhanstn222 (smh) mutant encodes the coiled-coil domain containing 103 protein (Ccdc103), a foxj1a regulated gene. Screening 146 unrelated PCD families identified patients in six families with reduced outer dynein arms, carrying mutations in CCDC103. Dynein arm assembly in smh mutant zebrafish was rescued by wild-type but not mutant human CCDC103. Chlamydomonas Ccdc103 functions as a tightly bound, axoneme-associated protein. The results identify Ccdc103 as a novel dynein arm attachment factor that when mutated causes Primary Ciliary Dyskinesia. PMID:22581229

Comparative assessment of single and joint effects of diuron and Irgarol 1051 on Arctic and temperate microalgae using chlorophyll a fluorescence imaging

KAUST Repository

Kottuparambil, Sreejith; Brown, Murray T.; Park, Jihae; Choi, Soyeon; Lee, Hojun; Choi, Han-Gu; Depuydt, Stephen; Han, Taejun

2017-01-01

Ship groundings and ice-breakers can cause pollution of the polar environment with antifouling biocides such as diuron and Irgarol 1051. The present study used pulse amplitude modulated fluorometry to compare single and joint toxicities of diuron and Irgarol 1051 on two freshwater taxa of microalgae (Chlorella and Chlamydomonas) originating from Arctic and temperate regions. 30min acute toxicity tests using chlorophyll a (Chl a) fluorescence revealed that Arctic strains of microalgae were more sensitive to herbicides than their temperate counterparts. Diuron and Irgarol 1051 had equal toxicities in the Arctic species, while Irgarol 1051 was more toxic (EC50=5.55–14.70μgL−1) than diuron (EC50=12.90–>40μgL−1) in the temperate species. Toxicity assessment of various mixtures of diuron and Irgarol 1051 revealed antagonistic, additive, and synergistic effects. Our data suggest that herbicides can adversely affect photosynthesis in Arctic microalgae at relatively low levels, and their impact can increase under complex mixture conditions.

Reduction in OER with LET: evidence supporting the ''oxygen-in-the-track'' hypothesis

International Nuclear Information System (INIS)

Bryant, P.E.

1976-01-01

To account for the reduction in OER with increasing LET which is observed for a wide variety of cell systems. Neary (1965) invoked the hypothesis that molecular oxygen is generated within the particle tracks of the more densely ionizing radiations. With the proviso that the product generated may be a species other than oxygen, but with similar sensitizing properties, produced by different qualities of radiation in two different organisms: the alga Chlamydomonas reinhardii and the bacterium Shigella flexneri were calculated. These effective concentrations should be the same for any given quality if the formation of the product is a function only of physico-chemical events occurring within the tracks of particles, and is independent of the biological material in which energy deposition is taking place. A prerequisite for the calculation of effective amounts of oxygen in tracks of ionizing particles is a knowledge of how radiosentivity varies with oxygen concentration at low LET

Mesocosm studies to assess acidity removal from acidic mine lakes through controlled eutrophication

International Nuclear Information System (INIS)

Fyson, A.; Nixdorf, B.; Kalin, M.; Steinberg, C.E.W.

1998-01-01

Flooded lignite pits (Tagebaurestseen) in Lusatia, Germany, are acidic (pH 2.5-4) with high concentrations of iron. Mesocosms (total volume 20 l) were set up with water and sediment from a Tagebaurestsee to assess the effects of phosphate and organic amendments under natural light and low temperature. Chemical and biological parameters were observed over a 9-month period. Phosphate rock addition resulted in sustained reduction in acidity in the water column and induced the growth of Chlamydomonas spp. (Chlorophyceae) near the water surface and Lepocinclis teres (Euglenophyceae) in a band above the sediment. Addition of potatoes to mesocosms resulted in the generation of near-anoxic conditions above the sediment, and phosphorus, ammonium and carbon (organic and inorganic) were released as the potatoes decomposed. A pH > 6 was attained with 5.1 g (dry weight) of potatoes and pH > 8 with 34 g (dry weight). In both mesocosms, more than 90% of total acidity was removed

Adaptability and variability of the cell functions to the environmental factors

Energy Technology Data Exchange (ETDEWEB)

Kikuchi, Tadatoshi [Kyoto Univ., Kumatori, Osaka (Japan). Research Reactor Inst.

1995-02-01

Adaptive phenomenon of the cells to the environmental factors is one of the most important functions of cells. In the initial research program, yeast, Saccharomyces cerevisiae, as model species of eukaryote was selected to use for the experiments and copper sulfate was adopted as one of the ideal environmental factors, and then adaptation mechanisms of yeast cells in the environment surrounded by copper ions were analyzed metabolically and morphologically. Furthermore, in the relationships between environmental factors and the cells, the researches performed were as follows: (1) Induced mutation in the extranuclear-inheritable system: Mutagenic effect of ethidium bromide on mitochondria and plastids. (2) Induction of gene expression by light exposure in the early development of chloroplast in Chlamydomonas reinhardi. (3) Some features of RNA and protein syntheses in thermophilic alga Cyanidium caldarium. (4) Satellite DNA of Ochromonas danica. (5) Analyses of cell functions using various kinds of radiations. (6) Novel methionine requirement of radiation resistant bacterium, Deinococcus radiodurans. (author).

Genomic analysis of organismal complexity in the multicellular green alga Volvox carteri

Energy Technology Data Exchange (ETDEWEB)

Prochnik, Simon E.; Umen, James; Nedelcu, Aurora; Hallmann, Armin; Miller, Stephen M.; Nishii, Ichiro; Ferris, Patrick; Kuo, Alan; Mitros, Therese; Fritz-Laylin, Lillian K.; Hellsten, Uffe; Chapman, Jarrod; Simakov, Oleg; Rensing, Stefan A.; Terry, Astrid; Pangilinan, Jasmyn; Kapitonov, Vladimir; Jurka, Jerzy; Salamov, Asaf; Shapiro, Harris; Schmutz, Jeremy; Grimwood, Jane; Lindquist, Erika; Lucas, Susan; Grigoriev, Igor V.; Schmitt, Rudiger; Kirk, David; Rokhsar, Daniel S.

2010-07-01

Analysis of the Volvox carteri genome reveals that this green alga's increased organismal complexity and multicellularity are associated with modifications in protein families shared with its unicellular ancestor, and not with large-scale innovations in protein coding capacity. The multicellular green alga Volvox carteri and its morphologically diverse close relatives (the volvocine algae) are uniquely suited for investigating the evolution of multicellularity and development. We sequenced the 138 Mb genome of V. carteri and compared its {approx}14,500 predicted proteins to those of its unicellular relative, Chlamydomonas reinhardtii. Despite fundamental differences in organismal complexity and life history, the two species have similar protein-coding potentials, and few species-specific protein-coding gene predictions. Interestingly, volvocine algal-specific proteins are enriched in Volvox, including those associated with an expanded and highly compartmentalized extracellular matrix. Our analysis shows that increases in organismal complexity can be associated with modifications of lineage-specific proteins rather than large-scale invention of protein-coding capacity.

Hypoxic survival requires a 2-on-2 hemoglobin in a process involving nitric oxide

Science.gov (United States)

Hemschemeier, Anja; Düner, Melis; Casero, David; Merchant, Sabeeha S.; Winkler, Martin; Happe, Thomas

2013-01-01

Hemoglobins are recognized today as a diverse family of proteins present in all kingdoms of life and performing multiple reactions beyond O2 chemistry. The physiological roles of most hemoglobins remain elusive. Here, we show that a 2-on-2 (“truncated”) hemoglobin, termed THB8, is required for hypoxic growth and the expression of anaerobic genes in Chlamydomonas reinhardtii. THB8 is 1 of 12 2-on-2 hemoglobins in this species. It belongs to a subclass within the 2-on-2 hemoglobin class I family whose members feature a remarkable variety of domain arrangements and lengths. Posttranscriptional silencing of the THB8 gene results in the mis-regulation of several genes and a growth defect under hypoxic conditions. The latter is intensified in the presence of an NO scavenger, which also impairs growth of wild-type cells. As recombinant THB8 furthermore reacts with NO, the results of this study indicate that THB8 is part of an NO-dependent signaling pathway. PMID:23754374

Acetate and bicarbonate assimilation and metabolite formation in Chlamydomonas reinhardtii: a 13C-NMR study.

Directory of Open Access Journals (Sweden)

Himanshu Singh

Full Text Available Cellular metabolite analyses by (13C-NMR showed that C. reinhardtii cells assimilate acetate at a faster rate in heterotrophy than in mixotrophy. While heterotrophic cells produced bicarbonate and CO2aq, mixotrophy cells produced bicarbonate alone as predominant metabolite. Experiments with singly (13C-labelled acetate ((13CH(3-COOH or CH(3-(13COOH supported that both the (13C nuclei give rise to bicarbonate and CO2(aq. The observed metabolite(s upon further incubation led to the production of starch and triacylglycerol (TAG in mixotrophy, whereas in heterotrophy the TAG production was minimal with substantial accumulation of glycerol and starch. Prolonged incubation up to eight days, without the addition of fresh acetate, led to an increased TAG production at the expense of bicarbonate, akin to that of nitrogen-starvation. However, such TAG production was substantially high in mixotrophy as compared to that in heterotrophy. Addition of mitochondrial un-coupler blocked the formation of bicarbonate and CO2(aq in heterotrophic cells, even though acetate uptake ensued. Addition of PSII-inhibitor to mixotrophic cells resulted in partial conversion of bicarbonate into CO2(aq, which were found to be in equilibrium. In an independent experiment, we have monitored assimilation of bicarbonate via photoautotrophy and found that the cells indeed produce starch and TAG at a much faster rate as compared to that in mixotrophy and heterotrophy. Further, we noticed that the accumulation of starch is relatively more as compared to TAG. Based on these observations, we suggest that acetate assimilation in C. reinhardtii does not directly lead to TAG formation but via bicarbonate/CO2(aq pathways. Photoautotrophic mode is found to be the best growth condition for the production of starch and TAG and starch in C. reinhardtii.

Regulation of the Chlamydomonas Cell Cycle by a Stable, Chromatin-Associated Retinoblastoma Tumor Suppressor Complex

Czech Academy of Sciences Publication Activity Database

Olson, B. J. S. C.; Oberholzer, M.; Li, Y.; Zones, J. M.; Kohli, H. S.; Bišová, Kateřina; Fang, S.; Ch.; Meisenhelder, J.; Hunter, T.; Umen, J. G.

2010-01-01

Roč. 22, č. 10 (2010), s. 3331-3347 ISSN 1040-4651 Institutional research plan: CEZ:AV0Z50200510 Keywords : GENOME-WIDE ANALYSIS * TRANSCRIPTION FACTOR * GENE-PRODUCT Subject RIV: EE - Microbiology, Virology Impact factor: 9.396, year: 2010

Role of the Rubisco Small Subunit

Energy Technology Data Exchange (ETDEWEB)

Spreitzer, Robert Joseph [Univ. of Nebraska, Lincoln, NE (United States)

2016-11-05

Ribulose-1,5-bisphosphate carboxylase/oxygenase (Rubisco) catalyzes the rate-limiting step of CO₂ fixation in photosynthesis. However, it is a slow enzyme, and O₂ competes with CO₂ at the active site. Oxygenation initiates the photorespiratory pathway, which also results in the loss of CO₂. If carboxylation could be increased or oxygenation decreased, an increase in net CO₂ fixation would be realized. Because Rubisco provides the primary means by which carbon enters all life on earth, there is much interest in engineering Rubisco to increase the production of food and renewable energy. Rubisco is located in the chloroplasts of plants, and it is comprised of two subunits. Much is known about the chloroplast-gene-encoded large subunit (rbcL gene), which contains the active site, but much less is known about the role of the nuclear-gene-encoded small subunit in Rubisco function (rbcS gene). Both subunits are coded by multiple genes in plants, which makes genetic engineering difficult. In the eukaryotic, green alga Chlamydomonas reinhardtii, it has been possible to eliminate all the Rubisco genes. These Rubisco-less mutants can be maintained by providing acetate as an alternative carbon source. In this project, focus has been placed on determining whether the small subunit might be a better genetic-engineering target for improving Rubisco. Analysis of a variable-loop structure (βA-βB loop) of the small subunit by genetic selection, directed mutagenesis, and construction of chimeras has shown that the small subunit can influence CO₂/O₂ specificity. X-ray crystal structures of engineered chimeric-loop enzymes have indicated that additional residues and regions of the small subunit may also contribute to Rubisco function. Structural dynamics of the small-subunit carboxyl terminus was also investigated. Alanine-scanning mutagenesis of the most-conserved small-subunit residues has identified a

Chloroplast DNA sequence of the green alga Oedogonium cardiacum (Chlorophyceae: Unique genome architecture, derived characters shared with the Chaetophorales and novel genes acquired through horizontal transfer

Directory of Open Access Journals (Sweden)

Lemieux Claude

2008-06-01

Full Text Available Abstract Background To gain insight into the branching order of the five main lineages currently recognized in the green algal class Chlorophyceae and to expand our understanding of chloroplast genome evolution, we have undertaken the sequencing of chloroplast DNA (cpDNA from representative taxa. The complete cpDNA sequences previously reported for Chlamydomonas (Chlamydomonadales, Scenedesmus (Sphaeropleales, and Stigeoclonium (Chaetophorales revealed tremendous variability in their architecture, the retention of only few ancestral gene clusters, and derived clusters shared by Chlamydomonas and Scenedesmus. Unexpectedly, our recent phylogenies inferred from these cpDNAs and the partial sequences of three other chlorophycean cpDNAs disclosed two major clades, one uniting the Chlamydomonadales and Sphaeropleales (CS clade and the other uniting the Oedogoniales, Chaetophorales and Chaetopeltidales (OCC clade. Although molecular signatures provided strong support for this dichotomy and for the branching of the Oedogoniales as the earliest-diverging lineage of the OCC clade, more data are required to validate these phylogenies. We describe here the complete cpDNA sequence of Oedogonium cardiacum (Oedogoniales. Results Like its three chlorophycean homologues, the 196,547-bp Oedogonium chloroplast genome displays a distinctive architecture. This genome is one of the most compact among photosynthetic chlorophytes. It has an atypical quadripartite structure, is intron-rich (17 group I and 4 group II introns, and displays 99 different conserved genes and four long open reading frames (ORFs, three of which are clustered in the spacious inverted repeat of 35,493 bp. Intriguingly, two of these ORFs (int and dpoB revealed high similarities to genes not usually found in cpDNA. At the gene content and gene order levels, the Oedogonium genome most closely resembles its Stigeoclonium counterpart. Characters shared by these chlorophyceans but missing in members

Morphological features of the species of the genus Chlamydomonas s.l. (Chlorophyta from various molecular clades

Directory of Open Access Journals (Sweden)

Maria N. Pavlovska

2012-03-01

Full Text Available The morphology of 78 authentic strains from 5 clades into culture condition was investigated. The complex of phenotype features was established. Such features as: type of mucilage and their origin, mucilage collapse under methylene blue, saving papilla and stigma in not motile stage, extracellular matrix formation inside cell wall, the way of sporangium break, pyrenoid and stigma habit before cell division, cell shape, chloroplast morphology. Diagnostic features for determination of taxa on clades level are discussed.

Excited-state kinetics of the carotenoid S//1 state in LHC II and two-photon excitation spectra of lutein and beta-carotene in solution Efficient Car S//1 yields Chl electronic energy transfer via hot S//1 states?

CERN Document Server

Walla, P J; Linden, Patricia A; Ohta, Kaoru

2002-01-01

The excited-state dynamics of the carotenoids (Car) in light- harvesting complex II (LHC II) of Chlamydomonas reinhardtii were studied by transient absorption measurements. The decay of the Car S //1 population ranges from similar to 200 fs to over 7 ps, depending on the excitation and detection wavelengths. In contrast, a 200 fs Car S//1 yields Chlorophyll (Chl) energy transfer component was the dominant time constant for our earlier two-photon fluorescence up- conversion measurements (Walla, P.J. ; et al. J. Phys. Chem. B 2000, 104, 4799-4806). We also present the two-photon excitation (TPE) spectra of lutein and beta-carotene in solution and compare them with the TPE spectrum of LHC II. The TPE-spectrum of LHC II has an onset much further to the blue and a width that is narrower than expected from comparison to the S//1 fluorescence of lutein and beta-carotene in solution. Different environments may affect the shape of the S//1 spectrum significantly. To explain the blue shift of the TPE spectrum and the d...

Ultraviolet light photobiology of the protozoan Tetrahymena pyriformis and chemical reactivation of DNA damage

International Nuclear Information System (INIS)

Wheeler, J.S.

1988-01-01

The tunable dye laser was developed in order to perform UV-B and UV-C (254-320 nm) action spectra studies on several different organisms. Using the laser, action spectra studies have been performed for Escherichia coli, Saccharomyces, Chlamydomonas, Caenorhabditis elegans, Paramecium, and Tetrahymena pyriformis. Studies generally indicate increasing LD 50 values with increasing wavelength. Two notable findings were made: (1) The action spectra does not follow the DNA absorption spectra at 280, 290 and 295 nm; (2) The repair competent/repair defective sensitization factor does not remain constant throughout the wavelength region. In addition it was found that the repair defective strain of E. coli, Bs-1, showed an increase in survival with increasing UV irradiation, at certain dose levels. Further experiments were designed to better characterize the reactivation. Tetrahymena were exposed to UV-C and reactivated with methyl methanesulfonate (MMS) and 4-nitro quinoline oxide (4-NQO). In both cases survival was seen to increase after chemical exposure. Likewise, UV-C was found to reactivate chemical damage (MMS)

SAS-6 engineering reveals interdependence between cartwheel and microtubules in determining centriole architecture.

Science.gov (United States)

Hilbert, Manuel; Noga, Akira; Frey, Daniel; Hamel, Virginie; Guichard, Paul; Kraatz, Sebastian H W; Pfreundschuh, Moritz; Hosner, Sarah; Flückiger, Isabelle; Jaussi, Rolf; Wieser, Mara M; Thieltges, Katherine M; Deupi, Xavier; Müller, Daniel J; Kammerer, Richard A; Gönczy, Pierre; Hirono, Masafumi; Steinmetz, Michel O

2016-04-01

Centrioles are critical for the formation of centrosomes, cilia and flagella in eukaryotes. They are thought to assemble around a nine-fold symmetric cartwheel structure established by SAS-6 proteins. Here, we have engineered Chlamydomonas reinhardtii SAS-6-based oligomers with symmetries ranging from five- to ten-fold. Expression of a SAS-6 mutant that forms six-fold symmetric cartwheel structures in vitro resulted in cartwheels and centrioles with eight- or nine-fold symmetries in vivo. In combination with Bld10 mutants that weaken cartwheel-microtubule interactions, this SAS-6 mutant produced six- to eight-fold symmetric cartwheels. Concurrently, the microtubule wall maintained eight- and nine-fold symmetries. Expressing SAS-6 with analogous mutations in human cells resulted in nine-fold symmetric centrioles that exhibited impaired length and organization. Together, our data suggest that the self-assembly properties of SAS-6 instruct cartwheel symmetry, and lead us to propose a model in which the cartwheel and the microtubule wall assemble in an interdependent manner to establish the native architecture of centrioles.

Metagenomic and satellite analyses of red snow in the Russian Arctic

Directory of Open Access Journals (Sweden)

Nao Hisakawa

2015-12-01

Full Text Available Cryophilic algae thrive in liquid water within snow and ice in alpine and polar regions worldwide. Blooms of these algae lower albedo (reflection of sunlight, thereby altering melting patterns (Kohshima, Seko & Yoshimura, 1993; Lutz et al., 2014; Thomas & Duval, 1995. Here metagenomic DNA analysis and satellite imaging were used to investigate red snow in Franz Josef Land in the Russian Arctic. Franz Josef Land red snow metagenomes confirmed that the communities are composed of the autotroph Chlamydomonas nivalis that is supporting a complex viral and heterotrophic bacterial community. Comparisons with white snow communities from other sites suggest that white snow and ice are initially colonized by fungal-dominated communities and then succeeded by the more complex C. nivalis-heterotroph red snow. Satellite image analysis showed that red snow covers up to 80% of the surface of snow and ice fields in Franz Josef Land and globally. Together these results show that C. nivalis supports a local food web that is on the rise as temperatures warm, with potential widespread impacts on alpine and polar environments worldwide.

POC1A truncation mutation causes a ciliopathy in humans characterized by primordial dwarfism.

Science.gov (United States)

Shaheen, Ranad; Faqeih, Eissa; Shamseldin, Hanan E; Noche, Ramil R; Sunker, Asma; Alshammari, Muneera J; Al-Sheddi, Tarfa; Adly, Nouran; Al-Dosari, Mohammed S; Megason, Sean G; Al-Husain, Muneera; Al-Mohanna, Futwan; Alkuraya, Fowzan S

2012-08-10

Primordial dwarfism (PD) is a phenotype characterized by profound growth retardation that is prenatal in onset. Significant strides have been made in the last few years toward improved understanding of the molecular underpinning of the limited growth that characterizes the embryonic and postnatal development of PD individuals. These include impaired mitotic mechanics, abnormal IGF2 expression, perturbed DNA-damage response, defective spliceosomal machinery, and abnormal replication licensing. In three families affected by a distinct form of PD, we identified a founder truncating mutation in POC1A. This gene is one of two vertebrate paralogs of POC1, which encodes one of the most abundant proteins in the Chlamydomonas centriole proteome. Cells derived from the index individual have abnormal mitotic mechanics with multipolar spindles, in addition to clearly impaired ciliogenesis. siRNA knockdown of POC1A in fibroblast cells recapitulates this ciliogenesis defect. Our findings highlight a human ciliopathy syndrome caused by deficiency of a major centriolar protein. Copyright © 2012 The American Society of Human Genetics. Published by Elsevier Inc. All rights reserved.

Effect of algae pigmentation on photobioreactor productivity and scale-up: A light transfer perspective

International Nuclear Information System (INIS)

Murphy, Thomas E.; Berberoglu, Halil

2011-01-01

This paper reports a numerical study coupling light transfer with photosynthetic rate models to determine the size and microorganism concentration of photobioreactors based on the pigmentation of algae to achieve maximum productivity. The wild strain Chlamydomonas reinhardtii and its transformant tla1 with 63% lower pigmentation are used as exemplary algae. First, empirical models of the specific photosynthetic rates were obtained from experimental data as a function of local irradiance using inverse methods. Then, these models were coupled with the radiative transfer equation (RTE) to predict both the local and total photosynthetic rates in a planar photobioreactor (PBR). The optical thickness was identified as the proper scaling parameter. The results indicated that under full sunlight corresponding to about 400 W/m 2 photosynthetically active irradiation, enhancement of PBR productivity up to 30% was possible with tla1. Moreover, under similar irradiation, optical thicknesses above 169 and 275 for the wild strain and tla1, respectively, did not further enhance PBR productivity. Based on these results guidelines are provided for maximizing PBR productivity from a light transport perspective.

Bio testing with micro algae in the pollution of environment with heavy metals

International Nuclear Information System (INIS)

Khudaverdiyev, S.R; Aliyev, E. Sh; Abdullayev, M.A; Khalilov

2010-01-01

Full text:Use of micro-algae is more expedient to determine how dangerous for living organisms the pollution of the environment with heavy metals. Photosynthesis is a process very sensitive to the impact of various factors. Photosynthesis is a process very sensitive to the impact of various factors. This process influences the changes in various non-photosynthetic solar energy losses of the indicator, which is the swallowing fluorescence especially in the late chordophone fluorescent (GF) parameters shows itself. The method of fluorescent indicator of photosynthetic membranes is based on the peculiar situation of the chordophone algae cells. Chlamydomonas Reinhardt by the research work presented in different environments micro-algae (Tries, taps), Fe, Ag and Au ions under the influence of changes of variable and delayed kinetics were studied fluorescent settings. In general, growth in the period of incubation experiments carried out in all the salts of the corresponding effect (increase or decrease) strengthened. At present, under the influence of radioactive pollution of the object to determine the degree of pollution due to changes in the initial work is being continued.

Photosynthetic biomanufacturing in green algae; production of recombinant proteins for industrial, nutritional, and medical uses.

Science.gov (United States)

Rasala, Beth A; Mayfield, Stephen P

2015-03-01

Recombinant proteins are widely used for industrial, nutritional, and medical applications. Green microalgae have attracted considerable attention recently as a biomanufacturing platform for the production of recombinant proteins for a number of reasons. These photosynthetic eukaryotic microorganisms are safe, scalable, easy to genetically modify through transformation, mutagenesis, or breeding, and inexpensive to grow. Many microalgae species are genetically transformable, but the green alga Chlamydomonas reinhardtii is the most widely used host for recombinant protein expression. An extensive suite of molecular genetic tools has been developed for C. reinhardtii over the last 25 years, including a fully sequenced genome, well-established methods for transformation, mutagenesis and breeding, and transformation vectors for high levels of recombinant protein accumulation and secretion. Here, we review recent successes in the development of C. reinhardtii as a biomanufacturing host for recombinant proteins, including antibodies and immunotoxins, hormones, industrial enzymes, an orally-active colostral protein for gastrointestinal health, and subunit vaccines. In addition, we review the biomanufacturing potential of other green algae from the genera Dunaliella and Chlorella.

Cloning and expression of isocitrate lyase from human round worm Strongyloides stercoralis

Directory of Open Access Journals (Sweden)

Siddiqui A.A.

2000-09-01

Full Text Available A full length cDNA (1463 bp encoding isocitrate lyase (EC 4.1.3.1 of Strongyloides stercoralis is described. The nucleotide sequence of this insert identified a cDNA coding for the isocitrate lyase. The conceptually translated amino acid sequence of the open reading frame for S. stercoralis isocitrate lyase encodes a 450 amino acid residue protein with an apparent molecular weight of 50 kDa and a predicted pl of 6.39. The sequence is 69 % A/T, reflecting a characteristic A/T codon bias of S. stercoralis. The amino acid sequence of S. stercoralis isocitrate lyase is compared with bifunctional glyoxylate cycle protein of Caenorhabditis elegans and isocitrate lyases from Chlamydomonas reinhardtii and Myxococcus xanthus. The full length cDNA of S. stercoralis was expressed in pRSET vector and bacteriophage T7 promoter based expression system. S. stercoralis lyase recombinant protein, purified via immobilized metal affinity chromatography, showed a molecular mass of 50 kDa on polyacrylamide gels. The role of isocitrate lyase in the glyoxylate cycle and energy metabolism of S. stercoralis is also discussed.

Optimized cryo-focused ion beam sample preparation aimed at in situ structural studies of membrane proteins.

Science.gov (United States)

Schaffer, Miroslava; Mahamid, Julia; Engel, Benjamin D; Laugks, Tim; Baumeister, Wolfgang; Plitzko, Jürgen M

2017-02-01

While cryo-electron tomography (cryo-ET) can reveal biological structures in their native state within the cellular environment, it requires the production of high-quality frozen-hydrated sections that are thinner than 300nm. Sample requirements are even more stringent for the visualization of membrane-bound protein complexes within dense cellular regions. Focused ion beam (FIB) sample preparation for transmission electron microscopy (TEM) is a well-established technique in material science, but there are only few examples of biological samples exhibiting sufficient quality for high-resolution in situ investigation by cryo-ET. In this work, we present a comprehensive description of a cryo-sample preparation workflow incorporating additional conductive-coating procedures. These coating steps eliminate the adverse effects of sample charging on imaging with the Volta phase plate, allowing data acquisition with improved contrast. We discuss optimized FIB milling strategies adapted from material science and each critical step required to produce homogeneously thin, non-charging FIB lamellas that make large areas of unperturbed HeLa and Chlamydomonas cells accessible for cryo-ET at molecular resolution. Copyright © 2016 Elsevier Inc. All rights reserved.

Metabolic Engineering of Escherichia coli for Producing Astaxanthin as the Predominant Carotenoid

Directory of Open Access Journals (Sweden)

Qian Lu

2017-09-01

Full Text Available Astaxanthin is a carotenoid of significant commercial value due to its superior antioxidant potential and wide applications in the aquaculture, food, cosmetic and pharmaceutical industries. A higher ratio of astaxanthin to the total carotenoids is required for efficient astaxanthin production. β-Carotene ketolase and hydroxylase play important roles in astaxanthin production. We first compared the conversion efficiency to astaxanthin in several β-carotene ketolases from Brevundimonas sp. SD212, Sphingomonas sp. DC18, Paracoccus sp. PC1, P. sp. N81106 and Chlamydomonas reinhardtii with the recombinant Escherichia coli cells that synthesize zeaxanthin due to the presence of the Pantoea ananatis crtEBIYZ. The B. sp. SD212 crtW and P. ananatis crtZ genes are the best combination for astaxanthin production. After balancing the activities of β-carotene ketolase and hydroxylase, an E. coli ASTA-1 that carries neither a plasmid nor an antibiotic marker was constructed to produce astaxanthin as the predominant carotenoid (96.6% with a specific content of 7.4 ± 0.3 mg/g DCW without an addition of inducer.

Algal Systems for Hydrogen Photoproduction

Energy Technology Data Exchange (ETDEWEB)

Ghirardi, Maria L [National Renewable Energy Lab. (NREL), Golden, CO (United States)

2015-10-08

The National Renewable Energy Laboratory (NREL), under the guidance of Drs. Michael Seibert (retired, Fellow Emeritus) and Maria Ghirardi (Fellow), led 15 years of research addressing the issue of algal H2 photoproduction. This project resulted in greatly increased rates and yields of algal hydrogen production; increased understanding of the H2 metabolism in the green alga, Chlamydomonas reinhardtii; expanded our knowledge of other physiological aspects relevant to sustained algal photosynthetic H2 production; led to the genetic identification, cloning and manipulation of algal hydrogenase genes; and contributed to a broader, fundamental understanding of the technical and scientific challenges to improving the conversion efficiencies in order to reach the U.S. Department of Energy’s Fuel Cell Technologies Office’s targets. Some of the tangible results are: (i) 64 publications and 6 patents, (ii) international visibility to NREL, (iii) reinvigoration of national and international biohydrogen research, and (iv) research progress that helped stimulate new funding from other DOE and non-DOE programs, including the AFOSR and the DOE Office of Science.

Enhanced FIB-SEM systems for large-volume 3D imaging

Science.gov (United States)

Xu, C Shan; Hayworth, Kenneth J; Lu, Zhiyuan; Grob, Patricia; Hassan, Ahmed M; García-Cerdán, José G; Niyogi, Krishna K; Nogales, Eva; Weinberg, Richard J; Hess, Harald F

2017-01-01

Focused Ion Beam Scanning Electron Microscopy (FIB-SEM) can automatically generate 3D images with superior z-axis resolution, yielding data that needs minimal image registration and related post-processing. Obstacles blocking wider adoption of FIB-SEM include slow imaging speed and lack of long-term system stability, which caps the maximum possible acquisition volume. Here, we present techniques that accelerate image acquisition while greatly improving FIB-SEM reliability, allowing the system to operate for months and generating continuously imaged volumes > 106 µm3. These volumes are large enough for connectomics, where the excellent z resolution can help in tracing of small neuronal processes and accelerate the tedious and time-consuming human proofreading effort. Even higher resolution can be achieved on smaller volumes. We present example data sets from mammalian neural tissue, Drosophila brain, and Chlamydomonas reinhardtii to illustrate the power of this novel high-resolution technique to address questions in both connectomics and cell biology. DOI: http://dx.doi.org/10.7554/eLife.25916.001 PMID:28500755

Hydrodynamic dispersion of microswimmers in suspension

Science.gov (United States)

Martin, Matthieu; Rafaï, Salima; Peyla, Philippe

2014-11-01

In our laboratory, we study hydrodynamics of suspensions of micro-swimmers. These micro-organisms are unicellular algae Chlamydomonas Rheinhardii which are able to swim by using their flagella. The swimming dynamics of these micro-swimmers can be seen as a random walk, in absence of any kind of interaction. In addition, these algae have the property of being phototactic, i.e. they swim towards the light. Combining this property with a hydrodynamic flow, we were able to reversibly separate algae from the rest of the fluid. But for sufficiently high volume fraction, these active particles interact with each other. We are now interested in how the coupling of hydrodynamic interactions between swimmers and phototaxis can modify the swimming dynamics at the scale of the suspension. To this aim, we conduct experiments in microfluidic devices to study the dispersion of the micro-organisms in a the liquid phase as a function of the volume fraction. We show that the dispersion of an assembly of puller type microswimmers is quantitatively affected by hydrodynamics interactions. Phd student.

A contribution to the study of plant development evolution based on gene co-expression networks

Directory of Open Access Journals (Sweden)

Francisco J. Romero-Campero

2013-08-01

Full Text Available Phototrophic eukaryotes are among the most successful organisms on Earth due to their unparalleled efficiency at capturing light energy and fixing carbon dioxide to produce organic molecules. A conserved and efficient network of light-dependent regulatory modules could be at the bases of this success. This regulatory system conferred early advantages to phototrophic eukaryotes that allowed for specialization, complex developmental processes and modern plant characteristics. We have studied light-dependent gene regulatory modules from algae to plants employing integrative-omics approaches based on gene co-expression networks. Our study reveals some remarkably conserved ways in which eukaryotic phototrophs deal with day length and light signaling. Here we describe how a family of Arabidopsis transcription factors involved in photoperiod response has evolved from a single algal gene according to the innovation, amplification and divergence theory of gene evolution by duplication. These modifications of the gene co-expression networks from the ancient unicellular green algae Chlamydomonas reinhardtii to the modern brassica Arabidopsis thaliana may hint on the evolution and specialization of plants and other organisms.

Stress and Protists: No life without stress.

Science.gov (United States)

Slaveykova, Vera; Sonntag, Bettina; Gutiérrez, Juan Carlos

2016-08-01

We report a summary of the symposium "Stress and Protists: No life without stress", which was held in September 2015 on the VII European Congress of Protistology in partnership with the International Society of Protistologists (Seville, Spain). We present an overview on general comments and concepts on cellular stress which can be also applied to any protist. Generally, various environmental stressors may induce similar cell responses in very different protists. Two main topics are reported in this manuscript: (i) metallic nanoparticles as environmental pollutants and stressors for aquatic protists, and (ii) ultraviolet radiation - induced stress and photoprotective strategies in ciliates. Model protists such as Chlamydomonas reinhardtii and Tetrahymena thermophila were used to assess stress caused by nanoparticles while stress caused by ultraviolet radiation was tested with free living planktonic ciliates as well as with the symbiont-bearing model ciliate Paramecium bursaria. For future studies, we suggest more intensive analyses on protist stress responses to specific environmental abiotic and/or biotic stressors at molecular and genetic levels up to ecological consequences and food web dynamics. Copyright © 2016 Elsevier GmbH. All rights reserved.

[Molecular, genetic and physiological analysis of photoinhibition and photosynthetic]. Progress report, June 1991--November 1992

Energy Technology Data Exchange (ETDEWEB)

1992-12-01

A major goal of this project is to use a combined molecular genetic, biochemical and physiological approach to understand the relationship between photosynthetic performance and the structure of the multifunctional D1 reaction center protein of Photosystem II encoded by the chloroplast psbA gene. Relative to other chloroplast proteins, turover of D1 is rapid and highly light dependent and de novo synthesis of D1 is required for a plant`s recovery from short term exposure to irradiances which induce photoinhibitory damage. These observations have led to models for a damage/repair cycle of PSII involving the targeted degradation and replacement of photodamaged D1. To investigate the effects of perturbing the D1 cycle on photosynthesis and autotrophic growth under high and low irradiance, we have examined the consequences of site-specific mutations of the psbA and 16S rRNA genes affecting synthesis, maturation and function/stability of the D1 protein introduced into the chloroplast genome of wildtype strain of the green alga Chlamydomonas reinhardtii using biolistic transformation.

[Molecular, genetic and physiological analysis of photoinhibition and photosynthetic

Energy Technology Data Exchange (ETDEWEB)

1992-01-01

A major goal of this project is to use a combined molecular genetic, biochemical and physiological approach to understand the relationship between photosynthetic performance and the structure of the multifunctional D1 reaction center protein of Photosystem II encoded by the chloroplast psbA gene. Relative to other chloroplast proteins, turover of D1 is rapid and highly light dependent and de novo synthesis of D1 is required for a plant's recovery from short term exposure to irradiances which induce photoinhibitory damage. These observations have led to models for a damage/repair cycle of PSII involving the targeted degradation and replacement of photodamaged D1. To investigate the effects of perturbing the D1 cycle on photosynthesis and autotrophic growth under high and low irradiance, we have examined the consequences of site-specific mutations of the psbA and 16S rRNA genes affecting synthesis, maturation and function/stability of the D1 protein introduced into the chloroplast genome of wildtype strain of the green alga Chlamydomonas reinhardtii using biolistic transformation.

The novel cytochrome c6 of chloroplasts: a case of evolutionary bricolage?

Science.gov (United States)

Howe, Christopher J; Schlarb-Ridley, Beatrix G; Wastl, Juergen; Purton, Saul; Bendall, Derek S

2006-01-01

Cytochrome c6 has long been known as a redox carrier of the thylakoid lumen of cyanobacteria and some eukaryotic algae that can substitute for plastocyanin in electron transfer. Until recently, it was widely accepted that land plants lack a cytochrome c6. However, a homologue of the protein has now been identified in several plant species together with an additional isoform in the green alga Chlamydomonas reinhardtii. This form of the protein, designated cytochrome c6A, differs from the 'conventional' cytochrome c6 in possessing a conserved insertion of 12 amino acids that includes two absolutely conserved cysteine residues. There are conflicting reports of whether cytochrome c6A can substitute for plastocyanin in photosynthetic electron transfer. The evidence for and against this is reviewed and the likely evolutionary history of cytochrome c6A is discussed. It is suggested that it has been converted from a primary role in electron transfer to one in regulation within the chloroplast, and is an example of evolutionary 'bricolage'.

Colonization of Snow by Microorganisms as Revealed Using Miniature Raman Spectrometers - Possibilities for Detecting Carotenoids of Psychrophiles on Mars?

Science.gov (United States)

Jehlička, Jan; Culka, Adam; Nedbalová, Linda

2016-12-01

We tested the potential of a miniaturized Raman spectrometer for use in field detection of snow algae pigments. A miniature Raman spectrometer, equipped with an excitation laser at 532 nm, allowed for the detection of carotenoids in cells of Chloromonas nivalis and Chlamydomonas nivalis at different stages of their life cycle. Astaxanthin, the major photoprotective pigment, was detected in algal blooms originating in snows at two alpine European sites that differed in altitude (Krkonoše Mts., Czech Republic, 1502 m a.s.l., and Ötztal Alps, Austria, 2790 m a.s.l.). Comparison is made with a common microalga exclusively producing astaxanthin (Haematococcus pluvialis). The handheld Raman spectrometer is a useful tool for fast and direct field estimations of the presence of carotenoids (mainly astaxanthin) within blooms of snow algae. Application of miniature Raman instruments as well as flight prototypes in areas where microbes are surviving under extreme conditions is an important stage in preparation for successful deployment of this kind of instrumentation in the framework of forthcoming astrobiological missions to Mars.

Temperature dependence of photosynthesis and thylakoid lipid composition in the red snow alga Chlamydomonas cf. nivalis (Chlotophyceae)

Czech Academy of Sciences Publication Activity Database

Lukeš, Martin; Procházková, L.; Shmidt, O.; Nedbalová, L.; Kaftan, David

2014-01-01

Roč. 89, č. 2 (2014), s. 303-315 ISSN 0168-6496 R&D Projects: GA MŠk ED2.1.00/03.0110 Grant - others:GAJU(CZ) 143/2013/P Institutional support: RVO:61388971 Keywords : electron transfer * snow * algae * photosynthesis Subject RIV: EE - Microbiology, Virology Impact factor: 3.568, year: 2014

Photoadduct Formation from the FMN Singlet Excited State in the LOV2 Domain of Chlamydomonas reinhardtii Phototropin

NARCIS (Netherlands)

Zhu, J.; Mathes, Tilo; Hontani, Y.; Alexandre, Maxime T A; Toh, K C; Hegemann, Peter; Kennis, John T M

2016-01-01

The two light, oxygen, and voltage domains of phototropin are blue-light photoreceptor domains that control various functions in plants and green algae. The key step of the light-driven reaction is the formation of a photoadduct between its FMN chromophore and a conserved cysteine, where the

X-ray and proton induced ultrastructural changes in Chlamydomonas reinhardi, with special reference to the dividing cell

International Nuclear Information System (INIS)

Gruber, H.E.

1976-01-01

Liquid cultures were exposed to 9000 R x-irradiation delivered at approximately 600 R/min. This produced 69 percent mortality in the 137c wild type strain and 71 percent mortality in the acetate-requiring strain ac-31. Irradiated and control cells were fixed for electron microscopic examination at intervals up to five days post exposure. Proton-irradiations using a positive ion Van de Graff accelerator were administered to monolayers of cells attached to Millipore filters. Irradiated and control cells were later resuspended and incubated in liquid culture medium. The dose rate was approximately 20 kilorad/second for thin targets with the dose monitored with a solid state detector. Distinctive fine structural responses were observed for the two kinds of radiation at the indicated exposure levels. Alterations affecting the nucleus were prominent after x-irradiation. Nuclei were observed in which non-nucleolar condensations and swollen nuclear envelopes were evident. Nuclear envelope rupture was noted when cells were in an advanced state of disorganization. Multiple nuclei per cell were also observed. Proton-irradiation often resulted in both live and dead daughter cells within the same mother cell wall. Changes in the chloroplast and mitochondria were seen after both types of irradiation. Two features absent in control cells were of special interest. Following x- and proton-irradiation, cells were observed which remained joined in configurations closely resembling division profiles or division conformations. Irradiated cells also possessed chromosomes and spindle fibers at a time not characteristic for such events in control cells

A dual system formed by the ARC and NR molybdoenzymes mediates nitrite-dependent NO production in Chlamydomonas.

Science.gov (United States)

Chamizo-Ampudia, Alejandro; Sanz-Luque, Emanuel; Llamas, Ángel; Ocaña-Calahorro, Francisco; Mariscal, Vicente; Carreras, Alfonso; Barroso, Juan B; Galván, Aurora; Fernández, Emilio

2016-10-01

Nitric oxide (NO) is a relevant signal molecule involved in many plant processes. However, the mechanisms and proteins responsible for its synthesis are scarcely known. In most photosynthetic organisms NO synthases have not been identified, and Nitrate Reductase (NR) has been proposed as the main enzymatic NO source, a process that in vitro is also catalysed by other molybdoenzymes. By studying transcriptional regulation, enzyme approaches, activity assays with in vitro purified proteins and in vivo and in vitro NO determinations, we have addressed the role of NR and Amidoxime Reducing Component (ARC) in the NO synthesis process. N\\R and ARC were intimately related both at transcriptional and activity level. Thus, arc mutants showed high NIA1 (NR gene) expression and NR activity. Conversely, mutants without active NR displayed an increased ARC expression in nitrite medium. Our results with nia1 and arc mutants and with purified enzymes support that ARC catalyses the NO production from nitrite taking electrons from NR and not from Cytb5-1/Cytb5-Reductase, the component partners previously described for ARC (proposed as NOFNiR, Nitric Oxide-Forming Nitrite Reductase). This NR-ARC dual system would be able to produce NO in the presence of nitrate, condition under which NR is unable to do it. © 2016 John Wiley & Sons Ltd.

Self-Replenishing Vascularized Fouling-Release Surfaces

Energy Technology Data Exchange (ETDEWEB)

Howell, C; Vu, TL; Lin, JJ; Kolle, S; Juthani, N; Watson, E; Weaver, JC; Alvarenga, J; Aizenberg, J

2014-08-13

Inspired by the long-term effectiveness of living antifouling materials, we have developed a method for the self-replenishment of synthetic biofouling-release surfaces. These surfaces are created by either molding or directly embedding 3D vascular systems into polydimethylsiloxane (PDMS) and filling them with a silicone oil to generate a nontoxic oil-infused material. When replenished with silicone oil from an outside source, these materials are capable of self-lubrication and continuous renewal of the interfacial fouling-release layer. Under accelerated lubricant loss conditions, fully infused vascularized samples retained significantly more lubricant than equivalent nonvascularized controls. Tests of lubricant-infused PDMS in static cultures of the infectious bacteria Staphylococcus aureus and Escherichia coli as well as the green microalgae Botryococcus braunii, Chlamydomonas reinhardtii, Dunaliella sauna, and Nannochloropsis oculata showed a significant reduction in biofilm adhesion compared to PDMS and glass controls containing no lubricant. Further experiments on vascularized versus nonvascularized samples that had been subjected to accelerated lubricant evaporation conditions for up to 48 h showed significantly less biofilm adherence on the vascularized surfaces. These results demonstrate the ability of an embedded lubricant-filled vascular network to improve the longevity of fouling-release surfaces.

Mycosporine-Like Amino Acids Promote Wound Healing through Focal Adhesion Kinase (FAK and Mitogen-Activated Protein Kinases (MAP Kinases Signaling Pathway in Keratinocytes

Directory of Open Access Journals (Sweden)

Yun-Hee Choi

2015-11-01

Full Text Available Mycosporine-like amino acids (MAAs are secondary metabolites found in diverse marine, freshwater, and terrestrial organisms. Evidence suggests that MAAs have several beneficial effects on skin homeostasis such as protection against UV radiation and reactive oxygen species (ROS. In addition, MAAs are also involved in the modulation of skin fibroblasts proliferation. However, the regulatory function of MAAs on wound repair in human skin is not yet clearly elucidated. To investigate the roles of MAAs on the wound healing process in human keratinocytes, three MAAs, Shinorine (SH, Mycosporine-glycine (M-Gly, and Porphyra (P334 were purified from Chlamydomonas hedlyei and Porphyra yezoensis. We found that SH, M-Gly, and P334 have significant effects on the wound healing process in human keratinocytes and these effects were mediated by activation of focal adhesion kinases (FAK, extracellular signal-regulated kinases (ERK, and c-Jun N-terminal kinases (JNK. These results suggest that MAAs accelerate wound repair by activating the FAK-MAPK signaling pathways. This study also indicates that MAAs can act as a new wound healing agent and further suggests that MAAs might be a novel biomaterial for wound healing therapies.

Mycosporine-Like Amino Acids Promote Wound Healing through Focal Adhesion Kinase (FAK) and Mitogen-Activated Protein Kinases (MAP Kinases) Signaling Pathway in Keratinocytes

Science.gov (United States)

Choi, Yun-Hee; Yang, Dong Joo; Kulkarni, Atul; Moh, Sang Hyun; Kim, Ki Woo

2015-01-01

Mycosporine-like amino acids (MAAs) are secondary metabolites found in diverse marine, freshwater, and terrestrial organisms. Evidence suggests that MAAs have several beneficial effects on skin homeostasis such as protection against UV radiation and reactive oxygen species (ROS). In addition, MAAs are also involved in the modulation of skin fibroblasts proliferation. However, the regulatory function of MAAs on wound repair in human skin is not yet clearly elucidated. To investigate the roles of MAAs on the wound healing process in human keratinocytes, three MAAs, Shinorine (SH), Mycosporine-glycine (M-Gly), and Porphyra (P334) were purified from Chlamydomonas hedlyei and Porphyra yezoensis. We found that SH, M-Gly, and P334 have significant effects on the wound healing process in human keratinocytes and these effects were mediated by activation of focal adhesion kinases (FAK), extracellular signal-regulated kinases (ERK), and c-Jun N-terminal kinases (JNK). These results suggest that MAAs accelerate wound repair by activating the FAK-MAPK signaling pathways. This study also indicates that MAAs can act as a new wound healing agent and further suggests that MAAs might be a novel biomaterial for wound healing therapies. PMID:26703626

The mother centriole plays an instructive role in defining cell geometry.

Directory of Open Access Journals (Sweden)

Jessica L Feldman

2007-06-01

Full Text Available Centriole positioning is a key step in establishment and propagation of cell geometry, but the mechanism of this positioning is unknown. The ability of pre-existing centrioles to induce formation of new centrioles at a defined angle relative to themselves suggests they may have the capacity to transmit spatial information to their daughters. Using three-dimensional computer-aided analysis of cell morphology in Chlamydomonas, we identify six genes required for centriole positioning relative to overall cell polarity, four of which have known sequences. We show that the distal portion of the centriole is critical for positioning, and that the centriole positions the nucleus rather than vice versa. We obtain evidence that the daughter centriole is unable to respond to normal positioning cues and relies on the mother for positional information. Our results represent a clear example of "cytotaxis" as defined by Sonneborn, and suggest that centrioles can play a key function in propagation of cellular geometry from one generation to the next. The genes documented here that are required for proper centriole positioning may represent a new class of ciliary disease genes, defects in which would be expected to cause disorganized ciliary position and impaired function.

Microbial Community Analysis of Colored Snow from an Alpine Snowfield in Northern Japan Reveals the Prevalence of Betaproteobacteria with Snow Algae.

Science.gov (United States)

Terashima, Mia; Umezawa, Kazuhiro; Mori, Shoichi; Kojima, Hisaya; Fukui, Manabu

2017-01-01

Psychrophilic algae blooms can be observed coloring the snow during the melt season in alpine snowfields. These algae are important primary producers on the snow surface environment, supporting the microbial community that coexists with algae, which includes heterotrophic bacteria and fungi. In this study, we analyzed the microbial community of green and red-colored snow containing algae from Mount Asahi, Japan. We found that Chloromonas spp. are the dominant algae in all samples analyzed, and Chlamydomonas is the second-most abundant genus in the red snow. For the bacterial community profile, species belonging to the subphylum Betaproteobacteria were frequently detected in both green and red snow, while members of the phylum Bacteroidetes were also prominent in red snow. Furthermore, multiple independently obtained strains of Chloromonas sp. from inoculates of red snow resulted in the growth of Betaproteobacteria with the alga and the presence of bacteria appears to support growth of the xenic algal cultures under laboratory conditions. The dominance of Betaproteobacteria in algae-containing snow in combination with the detection of Chloromonas sp. with Betaproteobacteria strains suggest that these bacteria can utilize the available carbon source in algae-rich environments and may in turn promote algal growth.

Evolution of green plants as deduced from 5S rRNA sequences.

Science.gov (United States)

Hori, H; Lim, B L; Osawa, S

1985-02-01

We have constructed a phylogenic tree for green plants by comparing 5S rRNA sequences. The tree suggests that the emergence of most of the uni- and multicellular green algae such as Chlamydomonas, Spirogyra, Ulva, and Chlorella occurred in the early stage of green plant evolution. The branching point of Nitella is a little earlier than that of land plants and much later than that of the above green algae, supporting the view that Nitella-like green algae may be the direct precursor to land plants. The Bryophyta and the Pteridophyta separated from each other after emergence of the Spermatophyta. The result is consistent with the view that the Bryophyta evolved from ferns by degeneration. In the Pteridophyta, Psilotum (whisk fern) separated first, and a little later Lycopodium (club moss) separated from the ancestor common to Equisetum (horsetail) and Dryopteris (fern). This order is in accordance with the classical view. During the Spermatophyta evolution, the gymnosperms (Cycas, Ginkgo, and Metasequoia have been studied here) and the angiosperms (flowering plants) separated, and this was followed by the separation of Metasequoia and Cycas (cycad)/Ginkgo (maidenhair tree) on one branch and various flowering plants on the other.

Zeaxanthin binds to light-harvesting complex stress-related protein to enhance nonphotochemical quenching in Physcomitrella patens.

Science.gov (United States)

Pinnola, Alberta; Dall'Osto, Luca; Gerotto, Caterina; Morosinotto, Tomas; Bassi, Roberto; Alboresi, Alessandro

2013-09-01

Nonphotochemical quenching (NPQ) dissipates excess energy to protect the photosynthetic apparatus from excess light. The moss Physcomitrella patens exhibits strong NPQ by both algal-type light-harvesting complex stress-related (LHCSR)-dependent and plant-type S subunit of Photosystem II (PSBS)-dependent mechanisms. In this work, we studied the dependence of NPQ reactions on zeaxanthin, which is synthesized under light stress by violaxanthin deepoxidase (VDE) from preexisting violaxanthin. We produced vde knockout (KO) plants and showed they underwent a dramatic reduction in thermal dissipation ability and enhanced photoinhibition in excess light conditions. Multiple mutants (vde lhcsr KO and vde psbs KO) showed that zeaxanthin had a major influence on LHCSR-dependent NPQ, in contrast with previous reports in Chlamydomonas reinhardtii. The PSBS-dependent component of quenching was less dependent on zeaxanthin, despite the near-complete violaxanthin to zeaxanthin exchange in LHC proteins. Consistent with this, we provide biochemical evidence that native LHCSR protein binds zeaxanthin upon excess light stress. These findings suggest that zeaxanthin played an important role in the adaptation of modern plants to the enhanced levels of oxygen and excess light intensity of land environments.

Zeaxanthin Binds to Light-Harvesting Complex Stress-Related Protein to Enhance Nonphotochemical Quenching in Physcomitrella patens[W

Science.gov (United States)

Pinnola, Alberta; Dall’Osto, Luca; Gerotto, Caterina; Morosinotto, Tomas; Bassi, Roberto; Alboresi, Alessandro

2013-01-01

Nonphotochemical quenching (NPQ) dissipates excess energy to protect the photosynthetic apparatus from excess light. The moss Physcomitrella patens exhibits strong NPQ by both algal-type light-harvesting complex stress-related (LHCSR)–dependent and plant-type S subunit of Photosystem II (PSBS)-dependent mechanisms. In this work, we studied the dependence of NPQ reactions on zeaxanthin, which is synthesized under light stress by violaxanthin deepoxidase (VDE) from preexisting violaxanthin. We produced vde knockout (KO) plants and showed they underwent a dramatic reduction in thermal dissipation ability and enhanced photoinhibition in excess light conditions. Multiple mutants (vde lhcsr KO and vde psbs KO) showed that zeaxanthin had a major influence on LHCSR-dependent NPQ, in contrast with previous reports in Chlamydomonas reinhardtii. The PSBS-dependent component of quenching was less dependent on zeaxanthin, despite the near-complete violaxanthin to zeaxanthin exchange in LHC proteins. Consistent with this, we provide biochemical evidence that native LHCSR protein binds zeaxanthin upon excess light stress. These findings suggest that zeaxanthin played an important role in the adaptation of modern plants to the enhanced levels of oxygen and excess light intensity of land environments. PMID:24014548

From models to crop species: caveats and solutions for translational metabolomics

Directory of Open Access Journals (Sweden)

Takayuki eTohge

2011-10-01

Full Text Available Although plant metabolomics is largely carried out on Arabidopsis it is essentially genome-independent, and thus potentially applicable to a wide range of species. However, transfer of between species, or even between different tissues of the same species, is not facile. This is because the reliability of protocols for harvesting, handling and analysis depends on the biological features and chemical composition of the plant tissue. In parallel with the diversification of model species it is important to establish good handling and analytic practice, in order to augment computational comparisons between tissues and species. LC-MS-based metabolomics is one of the powerful approaches for metabolite profiling. By using a combination of different extraction methods, separation columns and ion detection, a very wide range of metabolites can be analysed. However, its application requires careful attention to exclude potential pitfalls, including artifactual changes in metabolite levels during sample preparation and analytic errors due to ion-suppression. Here we provide case studies with two different LC-MS-based metabolomics platforms and four species (Arabidopsis thaliana, Chlamydomonas reinhardtii, Solanum lycopersicum and Oryza sativa that illustrate how such dangers can be detected and circumvented.

Comparative sensitivity of five species of macrophytes and six species of algae to atrazine, metribuzin, alachlor, and metolachlor

Science.gov (United States)

Fairchild, James F.; Ruessler, Shane; Carlson, A. Ron

1998-01-01

This study determined the relative sensitivity of five species of aquatic macrophytes and six species of algae to four commonly used herbicides (atrazine, metribuzin, alachlor, and metolachlor). Toxicity tests consisted of 96-h (duckweed and algae) or 14-d (submerged macrophytes) static exposures. The triazine herbicides (atrazine and metribuzin) were significantly more toxic to aquatic plants than were the acetanilide herbicides (alachlor and metolachlor). Toxicity studies ranked metribuzin > atrazine > alachlor > metolachlor in decreasing order of overall toxicity to aquatic plants. Relative sensitivities of macrophytes to these herbicides decreased in the order of Ceratophyllum > Najas > Elodea > Lemna > Myriophyllum. Relative sensitivities of algae to herbicides decreased in the order of Selenastrum > Chlorella > Chlamydomonas > Microcystis > Scenedesmus > Anabaena. Algae and macrophytes were of similar overall sensitivities to herbicides. Data indicated that Selenastrum, a commonly tested green alga, was generally more sensitive compared to other plant species. Lemna minor, a commonly tested floating vascular plant, was of intermediate sensitivity, and was fivefold less sensitive than Ceratophyllum, which was the most sensitive species tested. The results indicated that no species was consistently most sensitive, and that a suite of aquatic plant test species may be needed to perform accurate risk assessments of herbicides.

Comparison of nutrient removal capacity and biomass settleability of four high-potential microalgal species.

Science.gov (United States)

Su, Yanyan; Mennerich, Artur; Urban, Brigitte

2012-11-01

Four common used microalgae species were compared in terms of settleability, nutrient removal capacity and biomass productivity. After 1 month training, except cyanobacteria Phormidium sp., three green microalgae species, Chlamydomonas reinhardtii, Chlorella vulgaris and Scenedesmus rubescens, showed good settleability. The N and P removal efficiency was all above 99% within 7, 4, 6 and 6 days for N and 4, 2, 3 and 4 days for P, resulting in the N removal rates of 3.66±0.17, 6.39±0.20, 4.39±0.06 and 4.31±0.18 mg N/l/d and P removal rates of 0.56±0.07, 0.89±0.05, 0.76±0.09 and 0.60±0.05 mg P/l/d for Phormidium sp., C. reinhardtii, C. vulgaris and S. rubescens, respectively. Phormidium sp. had the lowest algal biomass productivity (2.71±0.7 g/m(2)/d) and the other three green microalgae showed higher algal biomass productivity (around 6 g/m(2)/d). Assimilation into biomass was the main removal mechanism for N and P. Copyright © 2012 Elsevier Ltd. All rights reserved.

Simultaneous effects of light intensity and phosphorus supply on the sterol content of phytoplankton.

Directory of Open Access Journals (Sweden)

Maike Piepho

Full Text Available Sterol profiles of microalgae and their change with environmental conditions are of great interest in ecological food web research and taxonomic studies alike. Here, we investigated effects of light intensity and phosphorus supply on the sterol content of phytoplankton and assessed potential interactive effects of these important environmental factors on the sterol composition of algae. We identified sterol contents of four common phytoplankton genera, Scenedesmus, Chlamydomonas, Cryptomonas and Cyclotella, and analysed the change in sterol content with varying light intensities in both a high-phosphorus and a low-phosphorus approach. Sterol contents increased significantly with increasing light in three out of four species. Phosphorus-limitation reversed the change of sterol content with light intensity, i.e., sterol content decreased with increasing light at low phosphorus supply. Generally sterol contents were lower in low-phosphorus cultures. In conclusion, both light and phosphorus conditions strongly affect the sterol composition of algae and hence should be considered in ecological and taxonomic studies investigating the biochemical composition of algae. Data suggest a possible sterol limitation of growth and reproduction of herbivorous crustacean zooplankton during summer when high light intensities and low phosphorus supply decrease sterol contents of algae.

Simple, rapid and cost-effective method for high quality nucleic acids extraction from different strains of Botryococcus braunii.

Directory of Open Access Journals (Sweden)

Byung-Hyuk Kim

Full Text Available This study deals with an effective nucleic acids extraction method from various strains of Botryococcus braunii which possesses an extensive extracellular matrix. A method combining freeze/thaw and bead-beating with heterogeneous diameter of silica/zirconia beads was optimized to isolate DNA and RNA from microalgae, especially from B. braunii. Eukaryotic Microalgal Nucleic Acids Extraction (EMNE method developed in this study showed at least 300 times higher DNA yield in all strains of B. braunii with high integrity and 50 times reduced working volume compared to commercially available DNA extraction kits. High quality RNA was also extracted using this method and more than two times the yield compared to existing methods. Real-time experiments confirmed the quality and quantity of the input DNA and RNA extracted using EMNE method. The method was also applied to other eukaryotic microalgae, such as diatoms, Chlamydomonas sp., Chlorella sp., and Scenedesmus sp. resulting in higher efficiencies. Cost-effectiveness analysis of DNA extraction by various methods revealed that EMNE method was superior to commercial kits and other reported methods by >15%. This method would immensely contribute to area of microalgal genomics.

Scanning electron microscope autoradiography of critical point dried biological samples

International Nuclear Information System (INIS)

Weiss, R.L.

1980-01-01

A technique has been developed for the localization of isotopes in the scanning electron microscope. Autoradiographic studies have been performed using a model system and a unicellular biflagellate alga. One requirement of this technique is that all manipulations be carried out on samples that are maintained in a liquid state. Observations of a source of radiation ( 125 I-ferritin) show that the nuclear emulsion used to detect radiation is active under these conditions. Efficiency measurement performed using 125 I-ferritin indicate that 125 I-SEM autoradiography is an efficient process that exhibits a 'dose dependent' response. Two types of labeling methods were used with cells, surface labeling with 125 I and internal labeling with 3 H. Silver grains appeared on labeled cells after autoradiography, removal of residual gelatin and critical point drying. The location of grains was examined on a flagellated green alga (Chlamydomonas reinhardi) capable of undergoing cell fusion. Fusion experiments using labeled and unlabeled cells indicate that 1. Labeling is specific for incorporated radioactivity; 2. Cell surface structure is preserved in SEM autoradiographs and 3. The technique appears to produce reliable autoradiographs. Thus scanning electron microscope autoradiography should provide a new and useful experimental approach

WD60/FAP163 is a dynein intermediate chain required for retrograde intraflagellar transport in cilia

Science.gov (United States)

Patel-King, Ramila S.; Gilberti, Renée M.; Hom, Erik F. Y.; King, Stephen M.

2013-01-01

Retrograde intraflagellar transport (IFT) is required for assembly of cilia. We identify a Chlamydomonas flagellar protein (flagellar-associated protein 163 [FAP163]) as being closely related to the D1bIC(FAP133) intermediate chain (IC) of the dynein that powers this movement. Biochemical analysis revealed that FAP163 is present in the flagellar matrix and is actively trafficked by IFT. Furthermore, FAP163 copurified with D1bIC(FAP133) and the LC8 dynein light chain, indicating that it is an integral component of the retrograde IFT dynein. To assess the functional role of FAP163, we generated an RNA interference knockdown of the orthologous protein (WD60) in planaria. The Smed-wd60(RNAi) animals had a severe ciliary assembly defect that dramatically compromised whole-organism motility. Most cilia were present as short stubs that had accumulated large quantities of IFT particle–like material between the doublet microtubules and the membrane. The few remaining approximately full-length cilia had a chaotic beat with a frequency reduced from 24 to ∼10 Hz. Thus WD60/FAP163 is a dynein IC that is absolutely required for retrograde IFT and ciliary assembly. PMID:23864713

Metabolic engineering of microalgal based biofuel production: prospects and challenges

Directory of Open Access Journals (Sweden)

Chiranjib eBanerjee

2016-03-01

Full Text Available The current scenario in renewable energy is focused on development of alternate and sustainable energy sources, amongst which microalgae stands as one of the promising feedstock for biofuel production. It is well known that microalgae generate much larger amounts of biofuels in a shorter time than other sources based on plant seeds. However, the greatest challenge in a transition to algae-based biofuel production is the various other complications involved in microalgal cultivation, its harvesting, concentration, drying and lipid extraction. Several green microalgae accumulate lipids, especially triacylglycerols (TAGs, which are main precursors in the production of lipid. The various aspects on metabolic pathway analysis of an oleaginous microalgae i.e. Chlamydomonas reinhardtii have elucidated some novel metabolically important genes and this enhances the lipid production in this microalgae. Adding to it, various other aspects in metabolic engineering using OptFlux and effectual bioprocess design also gives an interactive snapshot of enhancing lipid production which ultimately improvises the oil yield. This article reviews the current status of microalgal based technologies for biofuel production, bioreactor process design, flux analysis and it also provides various strategies to increase lipids accumulation via metabolic engineering.

Two-Dimensional Algal Collection and Assembly by Combining AC-Dielectrophoresis with Fluorescence Detection for Contaminant-Induced Oxidative Stress Sensing

Directory of Open Access Journals (Sweden)

Coralie Siebman

2015-06-01

Full Text Available An alternative current (AC dielectrophoretic lab-on-chip setup was evaluated as a rapid tool of capture and assembly of microalga Chlamydomonas reinhardtii in two-dimensional (2D close-packed arrays. An electric field of 100 V·cm−1, 100 Hz applied for 30 min was found optimal to collect and assemble the algae into single-layer structures of closely packed cells without inducing cellular oxidative stress. Combined with oxidative stress specific staining and fluorescence microscopy detection, the capability of using the 2D whole-cell assembly on-chip to follow the reactive oxygen species (ROS production and oxidative stress during short-term exposure to several environmental contaminants, including mercury, methylmercury, copper, copper oxide nanoparticles (CuO-NPs, and diuron was explored. The results showed significant increase of the cellular ROS when C. reinhardtii was exposed to high concentrations of methylmercury, CuO-NPs, and 10−5 M Cu. Overall, this study demonstrates the potential of combining AC-dielectrophoretically assembled two-dimensional algal structures with cell metabolic analysis using fluorescence staining, as a rapid analytical tool for probing the effect of contaminants in highly impacted environment.

Structure-Function Analysis of Chloroplast Proteins via Random Mutagenesis Using Error-Prone PCR.

Science.gov (United States)

Dumas, Louis; Zito, Francesca; Auroy, Pascaline; Johnson, Xenie; Peltier, Gilles; Alric, Jean

2018-06-01

Site-directed mutagenesis of chloroplast genes was developed three decades ago and has greatly advanced the field of photosynthesis research. Here, we describe a new approach for generating random chloroplast gene mutants that combines error-prone polymerase chain reaction of a gene of interest with chloroplast complementation of the knockout Chlamydomonas reinhardtii mutant. As a proof of concept, we targeted a 300-bp sequence of the petD gene that encodes subunit IV of the thylakoid membrane-bound cytochrome b 6 f complex. By sequencing chloroplast transformants, we revealed 149 mutations in the 300-bp target petD sequence that resulted in 92 amino acid substitutions in the 100-residue target subunit IV sequence. Our results show that this method is suited to the study of highly hydrophobic, multisubunit, and chloroplast-encoded proteins containing cofactors such as hemes, iron-sulfur clusters, and chlorophyll pigments. Moreover, we show that mutant screening and sequencing can be used to study photosynthetic mechanisms or to probe the mutational robustness of chloroplast-encoded proteins, and we propose that this method is a valuable tool for the directed evolution of enzymes in the chloroplast. © 2018 American Society of Plant Biologists. All rights reserved.

Role of benzoxazinones in allelopathy by rye (Secale cereale L.).

Science.gov (United States)

Barnes, J P; Putnam, A R

1987-04-01

Two phytotoxic compounds [2,4-dihydroxy-1,4(2H)-benzoxazin-3-one (DIBOA) and 2(3H)-benzoxazolinone (BOA)] were previously isolated and identified in 35-day-old greenhouse-grown rye shoot tissue. Both compounds were also detected by TLC in greenhouse-grown root and fieldgrown shoot tissue. The concentration of DIBOA varied in the tissues, with the greatest quantity detected in greenhouse-grown shoots. DIBOA and BOA were compared with β-phenyllactic acid (PLA) and β-hydroxybutyric acid (HBA) for activity on seed germination and seedling growth and were consistently more toxic than either compound. Dicot species tested, including lettuce (Lactuca sativa L.), tomato (Lycopersicon esculentum Mill.), and redroot pigweed (Amaranthus retroflexus L.), were 30% more sensitive than the monocots tested. Of the two benzoxazinone compounds, DIBOA was most toxic to seedling growth. DIBOA and BOA reduced chlorophyll production inChlamydomonas rheinhardtii Dangeard, by 50% at 7.5 × 10(-5) M and 1.0 × 10(-3) M, respectively. Both DIBOA and BOA inhibited emergence of barnyardgrass (Echinochloa crusgalli L. Beauv.), cress (Lepidium sativum L.), and lettuce when applied to soil, indicating their potential for allelopathic activity.

Electrokinetically controlled fluid injection into unicellular microalgae.

Science.gov (United States)

Zhou, Xuewen; Zhang, Xixi; Boualavong, Jonathan; Durney, Andrew R; Wang, Tonghui; Kirschner, Scott; Wentz, Michaela; Mukaibo, Hitomi

2017-10-01

Electrokinetically controlled microinjection is reported as an effective transport mechanism for microinjection into the wild-type strain of the widely studied model microalga Chlamydomonas reinhardtii. A microinjection system using glass capillary pipettes was developed to capture and impale the motile cells. To apply an electric field and induce electrokinetic flow (e.g., electrophoresis and electroosmosis), an electrode was inserted directly into the solution inside the impaling injection pipette and another electrode was inserted into the external cell media. The viability of the impaled cells was confirmed for more than an hour under 0.01 V using the fluorescein diacetate/propidium iodide dual fluorescent dye based assay. The viability was also found to increase almost logarithmically with decreasing voltage and to depend strongly on the solution within the injection pipette. Successful electrokinetic microinjection into cells was confirmed by both an increase in cell volume under an applied voltage and electric field dependent delivery of fluorescent fluorescein molecules into an impaled cell. Our study offers novel opportunities for quantitative delivery of biomolecules into microalgae and advancing the research and development of these organisms as biosynthetic factories. © 2017 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

Target of rapamycin (TOR) plays a critical role in triacylglycerol accumulation in microalgae.

Science.gov (United States)

Imamura, Sousuke; Kawase, Yasuko; Kobayashi, Ikki; Sone, Toshiyuki; Era, Atsuko; Miyagishima, Shin-Ya; Shimojima, Mie; Ohta, Hiroyuki; Tanaka, Kan

2015-10-01

Most microalgae produce triacylglycerol (TAG) under stress conditions such as nitrogen depletion, but the underlying molecular mechanism remains unclear. In this study, we focused on the role of target of rapamycin (TOR) in TAG accumulation. TOR is a serine/threonine protein kinase that is highly conserved and plays pivotal roles in nitrogen and other signaling pathways in eukaryotes. We previously constructed a rapamycin-susceptible Cyanidioschyzon merolae, a unicellular red alga, by expressing yeast FKBP12 protein to evaluate the results of TOR inhibition (Imamura et al. in Biochem Biophys Res Commun 439:264-269, 2013). By using this strain, we here report that rapamycin-induced TOR inhibition results in accumulation of cytoplasmic lipid droplets containing TAG. Transcripts for TAG synthesis-related genes, such as glycerol-3-phosphate acyltransferase and acyl-CoA:diacylglycerol acyltransferase (DGAT), were increased by rapamycin treatment. We also found that fatty acid synthase-dependent de novo fatty acid synthesis was required for the accumulation of lipid droplets. Induction of TAG and up-regulation of DGAT gene expression by rapamycin were similarly observed in the unicellular green alga, Chlamydomonas reinhardtii. These results suggest the general involvement of TOR signaling in TAG accumulation in divergent microalgae.

Probing Contaminant-Induced Alterations in Chlorophyll Fluorescence by AC-Dielectrophoresis-Based 2D-Algal Array

Directory of Open Access Journals (Sweden)

Coralie Siebman

2018-02-01

Full Text Available The investigation of contaminant impact on algae requires rapid and reliable cell collection and optical detection. The capability of alternative current (AC dielectrophoresis (DEP collection of whole cell arrays with combined fluorescence microscopy detection to follow the alterations of chlorophyll fluorescence during environmental contaminant exposure was explored. The application of an AC-field of 100 V cm−1, 100 Hz for 30 min to capture and immobilize the cells of green alga Chlamydomonas reinhardtii in two-dimensional (2D arrays does not induce changes in chlorophyll fluorescence. The results demonstrate that DEP-based 2D-arrays allow non-invasive detection of chlorophyll fluorescence change upon exposure to high concentrations of copper oxide nanoparticles and ionic copper. These results were in agreement with data obtained by flow cytometry used as a comparative method. The tool was also applied to follow the effect of a number of ubiquitous contaminants such as inorganic mercury, methylmercury, and diuron. However, a statistically significant short-term effect was observed only for mercury. Overall, DEP-based 2D-arrays of algal cells with fluorescence detection appear to be suitable for stain-free probing the effects on the photosynthetic microorganisms in highly polluted environment.

Hydrogen production from water: Recent advances in photosynthesis research

Energy Technology Data Exchange (ETDEWEB)

Greenbaum, E.; Lee, J.W. [Oak Ridge National Lab., TN (United States). Chemical Technology Div.

1997-12-31

The great potential of hydrogen production by microalgal water splitting is predicated on quantitative measurement of the algae`s hydrogen-producing capability, which is based on the following: (1) the photosynthetic unit size of hydrogen production; (2) the turnover time of photosynthetic hydrogen production; (3) thermodynamic efficiencies of conversion of light energy into the Gibbs free energy of molecular hydrogen; (4) photosynthetic hydrogen production from sea water using marine algae; (5) the potential for research advances using modern methods of molecular biology and genetic engineering to maximize hydrogen production. ORNL has shown that sustained simultaneous photoevolution of molecular hydrogen and oxygen can be performed with mutants of the green alga Chlamydomonas reinhardtii that lack a detectable level of the Photosystem I light reaction. This result is surprising in view of the standard two-light reaction model of photosynthesis and has interesting scientific and technological implications. This ORNL discovery also has potentially important implications for maximum thermodynamic conversion efficiency of light energy into chemical energy by green plant photosynthesis. Hydrogen production performed by a single light reaction, as opposed to two, implies a doubling of the theoretically maximum thermodynamic conversion efficiency from {approx}10% to {approx}20%.

Biodiesel production from wet microalgae feedstock using sequential wet extraction/transesterification and direct transesterification processes.

Science.gov (United States)

Chen, Ching-Lung; Huang, Chien-Chang; Ho, Kao-Chia; Hsiao, Ping-Xuan; Wu, Meng-Shan; Chang, Jo-Shu

2015-10-01

Although producing biodiesel from microalgae seems promising, there is still a lack of technology for the quick and cost-effective conversion of biodiesel from wet microalgae. This study was aimed to develop a novel microalgal biodiesel producing method, consisting of an open system of microwave disruption, partial dewatering (via combination of methanol treatment and low-speed centrifugation), oil extraction, and transesterification without the pre-removal of the co-solvent, using Chlamydomonas sp. JSC4 with 68.7 wt% water content as the feedstock. Direct transesterification with the disrupted wet microalgae was also conducted. The biomass content of the wet microalgae increased to 56.6 and 60.5 wt%, respectively, after microwave disruption and partial dewatering. About 96.2% oil recovery was achieved under the conditions of: extraction temperature, 45°C; hexane/methanol ratio, 3:1; extraction time, 80 min. Transesterification of the extracted oil reached 97.2% conversion within 15 min at 45°C and 6:1 solvent/methanol ratio with simultaneous Chlorophyll removal during the process. Nearly 100% biodiesel conversion was also obtained while conducting direct transesterification of the disrupted oil-bearing microalgal biomass. Copyright © 2015 Elsevier Ltd. All rights reserved.

LHCSR1 induces a fast and reversible pH-dependent fluorescence quenching in LHCII in Chlamydomonas reinhardtii cells

NARCIS (Netherlands)

Dinc, E.; Tian, L.; Roy, L.M.; Roth, R.; Goodenough, U.; Croce, R.

2016-01-01

To avoid photodamage, photosynthetic organisms are able to thermally dissipate the energy absorbed in excess in a process known as nonphotochemical quenching (NPQ). Although NPQ has been studied extensively, the major players and the mechanism of quenching remain debated. This is a result of the

Molecular mechanisms behind the adjustment of phototrophic light-harvesting and mixotrophic utilization of cellulosic carbon sources in Chlamydomonas reinhardtii

OpenAIRE

Blifernez-Klassen, Olga

2012-01-01

Plants, green algae and cyanobacteria perform photosynthetic conversion of sunlight into chemical energy in a permanently changing natural environment, where the efficient utilization of light and inorganic carbon represent the most critical factors. Photosynthetic organisms have developed different acclimation strategies to adapt changing light conditions and insufficient carbon source supply in order to survive and to assure optimal growth and protection. This thesis provides further insigh...

Consequences of state transitions on the structural and functional organization of Photosystem I in the green alga Chlamydomonas reinhardtii

NARCIS (Netherlands)

Drop, Bartlomiej; Yadav K.N., Sathish; Boekema, Egbert J.; Croce, Roberta

State transitions represent a photoacclimation process that regulates the light-driven photosynthetic reactions in response to changes in light quality/quantity. It balances the excitation between photosystem I (PSI) and II (PSII) by shuttling LHCII, the main light-harvesting complex of green algae