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Sample records for alga chlamydomonas sp

  1. Growth and lipid content at low temperature of Arctic alga Chlamydomonas sp. KNM0029C.

    PubMed

    Kim, Eun Jae; Jung, Woongsic; Lim, Suyoun; Kim, Sanghee; Han, Se Jong; Choi, Han-Gu

    2016-01-01

    Biodiesel produced from microalgae is a promising source of alternative energy. In winter, however, outdoor mass cultivation for biodiesel production is hampered by poor growth. Here, we report that Arctic Chlamydomonas sp. KNM0029C exhibits optimal growth at 4 °C and reaches densities up to 1.4 × 10(7) cells mL(-1). Lipid body formation in the alga was visualized through BODIPY 505/515 staining and fluorescence microscopy. The fatty acid methyl ester (FAME) production level of KNM0029C was 178.6 mg L(-1) culture and 2.3-fold higher than that of C. reinhardtii CC-125 at 4 °C. Analysis of the FAME content showed a predominance of polyunsaturated fatty acids such as C16:3, C18:2, C18:3, and C20:2. C18:3 fatty acids comprised the largest fraction (20.7%), and the content of polyunsaturated fatty acids (39.6%) was higher than that of saturated fatty acids (6.8%) at 4 °C. These results indicate that Chlamydomonas sp. KNM0029C, as a psychrophilic microalga, might represent a favorable source for biodiesel production in cold environments.

  2. Transcriptome-wide analysis of DEAD-box RNA helicase gene family in an Antarctic psychrophilic alga Chlamydomonas sp. ICE-L.

    PubMed

    Liu, Chenlin; Huang, Xiaohang

    2015-09-01

    DEAD-box RNA helicase family proteins have been identified in almost all living organisms. Some of them play a crucial role in adaptation to environmental changes and stress response, especially in the low-temperature acclimation in different kinds of organisms. Compared with the full swing study in plants and bacteria, the characters and functions of DEAD-box family proteins had not been surveyed in algae. To identify genes critical for freezing acclimation in algae, we screened DEAD-box RNA helicase genes from the transcriptome sequences of a psychrophilic microalga Chlamydomonas sp. ICE-L which was isolated from Antarctic sea ice. Totally 39 DEAD-box RNA helicase genes had been identified. Most of the DEAD-box RNA helicase have 1:1 homologous relationships in Chlamydomonas reinhardtii and Chlamydomonas sp. ICE-L with several exceptions. The homologous proteins in ICE-L to the helicases critical for cold or freezing tolerance in Arabidopsis thaliana had been identified based on phylogenetic comparison studies. The response of these helicase genes is not always identical in the Chlamydomonas sp. ICE-L and Arabidopsis under the same low-temperature treatment. The expression of several DEAD-box RNA helicase genes including CiRH5, CiRH25, CiRH28, and CiRH55 were significantly up-regulated under freezing treatment of ICE-L and their function in freezing acclimation of ICE-L deserved further investigation.

  3. Long-term experiment on physiological responses to synergetic effects of ocean acidification and photoperiod in the Antarctic sea ice algae Chlamydomonas sp. ICE-L.

    PubMed

    Xu, Dong; Wang, Yitao; Fan, Xiao; Wang, Dongsheng; Ye, Naihao; Zhang, Xiaowen; Mou, Shanli; Guan, Zheng; Zhuang, Zhimeng

    2014-07-15

    Studies on ocean acidification have mostly been based on short-term experiments of low latitude with few investigations of the long-term influence on sea ice communities. Here, the combined effects of ocean acidification and photoperiod on the physiological response of the Antarctic sea ice microalgae Chlamydomonas sp. ICE-L were examined. There was a general increase in growth, PSII photosynthetic parameters, and N and P uptake in continuous light, compared to those exposed to regular dark and light cycles. Elevated pCO2 showed no consistent effect on growth rate (p=0.8) and N uptake (p=0.38) during exponential phrase, depending on the photoperiod but had a positive effect on PSII photosynthetic capacity and P uptake. Continuous dark reduced growth, photosynthesis, and nutrient uptake. Moreover, intracellular lipid, mainly in the form of PUFA, was consumed at 80% and 63% in low and high pCO2 in darkness. However, long-term culture under high pCO2 gave a more significant inhibition of growth and Fv/Fm to high light stress. In summary, ocean acidification may have significant effects on Chlamydomonas sp. ICE-L survival in polar winter. The current study contributes to an understanding of how a sea ice algae-based community may respond to global climate change at high latitudes.

  4. Dynamic metabolic profiling together with transcription analysis reveals salinity-induced starch-to-lipid biosynthesis in alga Chlamydomonas sp. JSC4.

    PubMed

    Ho, Shih-Hsin; Nakanishi, Akihito; Kato, Yuichi; Yamasaki, Hiroaki; Chang, Jo-Shu; Misawa, Naomi; Hirose, Yuu; Minagawa, Jun; Hasunuma, Tomohisa; Kondo, Akihiko

    2017-04-04

    Biodiesel production using microalgae would play a pivotal role in satisfying future global energy demands. Understanding of lipid metabolism in microalgae is important to isolate oleaginous strain capable of overproducing lipids. It has been reported that reducing starch biosynthesis can enhance lipid accumulation. However, the metabolic mechanism controlling carbon partitioning from starch to lipids in microalgae remains unclear, thus complicating the genetic engineering of algal strains. We here used "dynamic" metabolic profiling and essential transcription analysis of the oleaginous green alga Chlamydomonas sp. JSC4 for the first time to demonstrate the switching mechanisms from starch to lipid synthesis using salinity as a regulator, and identified the metabolic rate-limiting step for enhancing lipid accumulation (e.g., pyruvate-to-acetyl-CoA). These results, showing salinity-induced starch-to-lipid biosynthesis, will help increase our understanding of dynamic carbon partitioning in oleaginous microalgae. Moreover, we successfully determined the changes of several key lipid-synthesis-related genes (e.g., acetyl-CoA carboxylase, pyruvate decarboxylase, acetaldehyde dehydrogenase, acetyl-CoA synthetase and pyruvate ferredoxin oxidoreductase) and starch-degradation related genes (e.g., starch phosphorylases), which could provide a breakthrough in the marine microalgal production of biodiesel.

  5. Dynamic metabolic profiling together with transcription analysis reveals salinity-induced starch-to-lipid biosynthesis in alga Chlamydomonas sp. JSC4

    PubMed Central

    Ho, Shih-Hsin; Nakanishi, Akihito; Kato, Yuichi; Yamasaki, Hiroaki; Chang, Jo-Shu; Misawa, Naomi; Hirose, Yuu; Minagawa, Jun; Hasunuma, Tomohisa; Kondo, Akihiko

    2017-01-01

    Biodiesel production using microalgae would play a pivotal role in satisfying future global energy demands. Understanding of lipid metabolism in microalgae is important to isolate oleaginous strain capable of overproducing lipids. It has been reported that reducing starch biosynthesis can enhance lipid accumulation. However, the metabolic mechanism controlling carbon partitioning from starch to lipids in microalgae remains unclear, thus complicating the genetic engineering of algal strains. We here used “dynamic” metabolic profiling and essential transcription analysis of the oleaginous green alga Chlamydomonas sp. JSC4 for the first time to demonstrate the switching mechanisms from starch to lipid synthesis using salinity as a regulator, and identified the metabolic rate-limiting step for enhancing lipid accumulation (e.g., pyruvate-to-acetyl-CoA). These results, showing salinity-induced starch-to-lipid biosynthesis, will help increase our understanding of dynamic carbon partitioning in oleaginous microalgae. Moreover, we successfully determined the changes of several key lipid-synthesis-related genes (e.g., acetyl-CoA carboxylase, pyruvate decarboxylase, acetaldehyde dehydrogenase, acetyl-CoA synthetase and pyruvate ferredoxin oxidoreductase) and starch-degradation related genes (e.g., starch phosphorylases), which could provide a breakthrough in the marine microalgal production of biodiesel. PMID:28374798

  6. Validation of housekeeping genes for gene expression studies in an ice alga Chlamydomonas during freezing acclimation.

    PubMed

    Liu, Chenlin; Wu, Guangting; Huang, Xiaohang; Liu, Shenghao; Cong, Bailin

    2012-05-01

    Antarctic ice alga Chlamydomonas sp. ICE-L can endure extreme low temperature and high salinity stress under freezing conditions. To elucidate the molecular acclimation mechanisms using gene expression analysis, the expression stabilities of ten housekeeping genes of Chlamydomonas sp. ICE-L during freezing stress were analyzed. Some discrepancies were detected in the ranking of the candidate reference genes between geNorm and NormFinder programs, but there was substantial agreement between the groups of genes with the most and the least stable expression. RPL19 was ranked as the best candidate reference genes. Pairwise variation (V) analysis indicated the combination of two reference genes was sufficient for qRT-PCR data normalization under the experimental conditions. Considering the co-regulation between RPL19 and RPL32 (the most stable gene pairs given by geNorm program), we propose that the mean data rendered by RPL19 and GAPDH (the most stable gene pairs given by NormFinder program) be used to normalize gene expression values in Chlamydomonas sp. ICE-L more accurately. The example of FAD3 gene expression calculation demonstrated the importance of selecting an appropriate category and number of reference genes to achieve an accurate and reliable normalization of gene expression during freeze acclimation in Chlamydomonas sp. ICE-L.

  7. Homogentisate phytyltransferase from the unicellular green alga Chlamydomonas reinhardtii.

    PubMed

    Gálvez-Valdivieso, Gregorio; Cardeñosa, Rosa; Pineda, Manuel; Aguilar, Miguel

    2015-09-01

    Homogentisate phytyltransferase (HPT) (EC 2.5.1.-) catalyzes the first committed step of tocopherol biosynthesis in all photosynthetic organisms. This paper presents the molecular characterization and expression analysis of HPT1 gene, and a study on the accumulation of tocopherols under different environmental conditions in the unicellular green alga Chlamydomonas reinhardtii. The Chlamydomonas HPT1 protein conserves all the prenylphosphate- and divalent cation-binding sites that are found in polyprenyltransferases and all the amino acids that are essential for its catalytic activity. Its hydrophobicity profile confirms that HPT is a membrane-bound protein. Chlamydomonas genomic DNA analysis suggests that HPT is encoded by a single gene, HPT1, whose promoter region contains multiple motifs related to regulation by jasmonate, abscisic acid, low temperature and light, and an ATCTA motif presents in genes involved in tocopherol biosynthesis and some photosynthesis-related genes. Expression analysis revealed that HPT1 is strongly regulated by dark and low-temperature. Under the same treatments, α-tocopherol increased in cultures exposed to darkness or heat, whereas γ-tocopherol did it in low temperature. The regulatory expression pattern of HPT1 and the changes of tocopherol abundance support the idea that different tocopherols play specific functions, and suggest a role for γ-tocopherol in the adaptation to growth under low-temperature.

  8. Resolving the phylogenetic relationship between Chlamydomonas sp. UWO 241 and Chlamydomonas raudensis sag 49.72 (Chlorophyceae) with nuclear and plastid DNA sequences.

    PubMed

    Possmayer, Marc; Gupta, Rajesh K; Szyszka-Mroz, Beth; Maxwell, Denis P; Lachance, Marc-André; Hüner, Norman P A; Smith, David Roy

    2016-04-01

    The Antarctic psychrophilic green alga Chlamy-domonas sp. UWO 241 is an emerging model for studying microbial adaptation to polar environments. However, little is known about its evolutionary history and its phylogenetic relationship with other chlamydomonadalean algae is equivocal. Here, we attempt to clarify the phylogenetic position of UWO 241, specifically with respect to Chlamydomonas rau-densis SAG 49.72. Contrary to a previous report, we show that UWO 241 is a distinct species from SAG 49.72. Our phylogenetic analyses of nuclear and plastid DNA sequences reveal that UWO 241 represents a unique lineage within the Moewusinia clade (sensu Nakada) of the Chlamydomonadales (Chlorophyceae, Chlorophyta), closely affiliated to the marine species Chlamydomonas parkeae SAG 24.89.

  9. Photosynthetic H2 metabolism in Chlamydomonas reinhardtii (unicellular green algae).

    PubMed

    Melis, Anastasios

    2007-10-01

    Unicellular green algae have the ability to operate in two distinctly different environments (aerobic and anaerobic), and to photosynthetically generate molecular hydrogen (H2). A recently developed metabolic protocol in the green alga Chlamydomonas reinhardtii permitted separation of photosynthetic O2-evolution and carbon accumulation from anaerobic consumption of cellular metabolites and concomitant photosynthetic H2-evolution. The H2 evolution process was induced upon sulfate nutrient deprivation of the cells, which reversibly inhibits photosystem-II and O2-evolution in their chloroplast. In the absence of O2, and in order to generate ATP, green algae resorted to anaerobic photosynthetic metabolism, evolved H2 in the light and consumed endogenous substrate. This study summarizes recent advances on green algal hydrogen metabolism and discusses avenues of research for the further development of this method. Included is the mechanism of a substantial tenfold starch accumulation in the cells, observed promptly upon S-deprivation, and the regulated starch and protein catabolism during the subsequent H2-evolution. Also discussed is the function of a chloroplast envelope-localized sulfate permease, and the photosynthesis-respiration relationship in green algae as potential tools by which to stabilize and enhance H2 metabolism. In addition to potential practical applications of H2, approaches discussed in this work are beginning to address the biochemistry of anaerobic H2 photoproduction, its genes, proteins, regulation, and communication with other metabolic pathways in microalgae. Photosynthetic H2 production by green algae may hold the promise of generating a renewable fuel from nature's most plentiful resources, sunlight and water. The process potentially concerns global warming and the question of energy supply and demand.

  10. Diversity in photosynthetic electron transport under [CO2]-limitation: the cyanobacterium Synechococcus sp. PCC 7002 and green alga Chlamydomonas reinhardtii drive an O2-dependent alternative electron flow and non-photochemical quenching of chlorophyll fluorescence during CO2-limited photosynthesis.

    PubMed

    Shimakawa, Ginga; Akimoto, Seiji; Ueno, Yoshifumi; Wada, Ayumi; Shaku, Keiichiro; Takahashi, Yuichiro; Miyake, Chikahiro

    2016-12-01

    Some cyanobacteria, but not all, experience an induction of alternative electron flow (AEF) during CO2-limited photosynthesis. For example, Synechocystis sp. PCC 6803 (S. 6803) exhibits AEF, but Synechococcus elongatus sp. PCC 7942 does not. This difference is due to the presence of flavodiiron 2 and 4 proteins (FLV2/4) in S. 6803, which catalyze electron donation to O2. In this study, we observed a low-[CO2] induced AEF in the marine cyanobacterium Synechococcus sp. PCC 7002 that lacks FLV2/4. The AEF shows high affinity for O2, compared with AEF mediated by FLV2/4 in S. 6803, and can proceed under extreme low [O2] (about a few µM O2). Further, the transition from CO2-saturated to CO2-limited photosynthesis leads a preferential excitation of PSI to PSII and increased non-photochemical quenching of chlorophyll fluorescence. We found that the model green alga Chlamydomonas reinhardtii also has an O2-dependent AEF showing the same affinity for O2 as that in S. 7002. These data represent the diverse molecular mechanisms to drive AEF in cyanobacteria and green algae. In this paper, we further discuss the diversity, the evolution, and the physiological function of strategy to CO2-limitation in cyanobacterial and green algal photosynthesis.

  11. How the green alga Chlamydomonas reinhardtii keeps time.

    PubMed

    Schulze, Thomas; Prager, Katja; Dathe, Hannes; Kelm, Juliane; Kiessling, Peter; Mittag, Maria

    2010-08-01

    The unicellular green alga Chlamydomonas reinhardtii has two flagella and a primitive visual system, the eyespot apparatus, which allows the cell to phototax. About 40 years ago, it was shown that the circadian clock controls its phototactic movement. Since then, several circadian rhythms such as chemotaxis, cell division, UV sensitivity, adherence to glass, or starch metabolism have been characterized. The availability of its entire genome sequence along with homology studies and the analysis of several sub-proteomes render C. reinhardtii as an excellent eukaryotic model organism to study its circadian clock at different levels of organization. Previous studies point to several potential photoreceptors that may be involved in forwarding light information to entrain its clock. However, experimental data are still missing toward this end. In the past years, several components have been functionally characterized that are likely to be part of the oscillatory machinery of C. reinhardtii since alterations in their expression levels or insertional mutagenesis of the genes resulted in defects in phase, period, or amplitude of at least two independent measured rhythms. These include several RHYTHM OF CHLOROPLAST (ROC) proteins, a CONSTANS protein (CrCO) that is involved in parallel in photoperiodic control, as well as the two subunits of the circadian RNA-binding protein CHLAMY1. The latter is also tightly connected to circadian output processes. Several candidates including a significant number of ROCs, CrCO, and CASEIN KINASE1 whose alterations of expression affect the circadian clock have in parallel severe effects on the release of daughter cells, flagellar formation, and/or movement, indicating that these processes are interconnected in C. reinhardtii. The challenging task for the future will be to get insights into the clock network and to find out how the clock-related factors are functionally connected. In this respect, system biology approaches will certainly

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

    DOE PAGES

    Therien, Jesse B.; Zadvornyy, Oleg A.; Posewitz, Matthew C.; ...

    2014-10-18

    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. We demonstrate the growth of C. reinhardtii on acetate provided by mutant strains of the cyanobacterium Synechococcus sp. PCC7002.

  13. Inhibition of target of rapamycin signaling by rapamycin in the unicellular green alga Chlamydomonas reinhardtii.

    PubMed

    Crespo, José L; Díaz-Troya, Sandra; Florencio, Francisco J

    2005-12-01

    The macrolide rapamycin specifically binds the 12-kD FK506-binding protein (FKBP12), and this complex potently inhibits the target of rapamycin (TOR) kinase. The identification of TOR in Arabidopsis (Arabidopsis thaliana) revealed that TOR is conserved in photosynthetic eukaryotes. However, research on TOR signaling in plants has been hampered by the natural resistance of plants to rapamycin. Here, we report TOR inactivation by rapamycin treatment in a photosynthetic organism. We identified and characterized TOR and FKBP12 homologs in the unicellular green alga Chlamydomonas reinhardtii. Whereas growth of wild-type Chlamydomonas cells is sensitive to rapamycin, cells lacking FKBP12 are fully resistant to the drug, indicating that this protein mediates rapamycin action to inhibit cell growth. Unlike its plant homolog, Chlamydomonas FKBP12 exhibits high affinity to rapamycin in vivo, which was increased by mutation of conserved residues in the drug-binding pocket. Furthermore, pull-down assays demonstrated that TOR binds FKBP12 in the presence of rapamycin. Finally, rapamycin treatment resulted in a pronounced increase of vacuole size that resembled autophagic-like processes. Thus, our findings suggest that Chlamydomonas cell growth is positively controlled by a conserved TOR kinase and establish this unicellular alga as a useful model system for studying TOR signaling in photosynthetic eukaryotes.

  14. Production of Recombinant Proteins in the Chloroplast of the Green Alga Chlamydomonas reinhardtii.

    PubMed

    Guzmán-Zapata, Daniel; Macedo-Osorio, Karla Soledad; Almaraz-Delgado, Alma Lorena; Durán-Figueroa, Noé; Badillo-Corona, Jesus Agustín

    2016-01-01

    Chloroplast transformation in the green algae Chlamydomonas reinhardtii can be used for the production of valuable recombinant proteins. Here, we describe chloroplast transformation of C. reinhardtii followed by protein detection. Genes of interest integrate stably by homologous recombination into the chloroplast genome following introduction by particle bombardment. Genes are inherited and expressed in lines recovered after selection in the presence of an antibiotic. Recombinant proteins can be detected by conventional techniques like immunoblotting and purified from liquid cultures.

  15. Predicting the Physiological Role of Circadian Metabolic Regulation in the Green Alga Chlamydomonas reinhardtii

    PubMed Central

    Voytsekh, Olga; Mittag, Maria; Schuster, Stefan

    2011-01-01

    Although the number of reconstructed metabolic networks is steadily growing, experimental data integration into these networks is still challenging. Based on elementary flux mode analysis, we combine sequence information with metabolic pathway analysis and include, as a novel aspect, circadian regulation. While minimizing the need of assumptions, we are able to predict changes in the metabolic state and can hypothesise on the physiological role of circadian control in nitrogen metabolism of the green alga Chlamydomonas reinhardtii. PMID:21887226

  16. Respiratory-deficient mutants of the unicellular green alga Chlamydomonas: a review.

    PubMed

    Salinas, Thalia; Larosa, Véronique; Cardol, Pierre; Maréchal-Drouard, Laurence; Remacle, Claire

    2014-05-01

    Genetic manipulation of the unicellular green alga Chlamydomonas reinhardtii is straightforward. Nuclear genes can be interrupted by insertional mutagenesis or targeted by RNA interference whereas random or site-directed mutagenesis allows the introduction of mutations in the mitochondrial genome. This, combined with a screen that easily allows discriminating respiratory-deficient mutants, makes Chlamydomonas a model system of choice to study mitochondria biology in photosynthetic organisms. Since the first description of Chlamydomonas respiratory-deficient mutants in 1977 by random mutagenesis, many other mutants affected in mitochondrial components have been characterized. These respiratory-deficient mutants increased our knowledge on function and assembly of the respiratory enzyme complexes. More recently some of these mutants allowed the study of mitochondrial gene expression processes poorly understood in Chlamydomonas. In this review, we update the data concerning the respiratory components with a special focus on the assembly factors identified on other organisms. In addition, we make an inventory of different mitochondrial respiratory mutants that are inactivated either on mitochondrial or nuclear genes.

  17. Ecophysiology, secondary pigments and ultrastructure of Chlainomonas sp. (Chlorophyta) from the European Alps compared with Chlamydomonas nivalis forming red snow

    PubMed Central

    Remias, Daniel; Pichrtová, Martina; Pangratz, Marion; Lütz, Cornelius; Holzinger, Andreas

    2016-01-01

    Red snow is a well-known phenomenon caused by microalgae thriving in alpine and polar regions during the melting season. The ecology and biodiversity of these organisms, which are adapted to low temperatures, high irradiance and freeze–thaw events, are still poorly understood. We compared two different snow habitats containing two different green algal genera in the European Alps, namely algae blooming in seasonal rock-based snowfields (Chlamydomonas nivalis) and algae dominating waterlogged snow bedded over ice (Chlainomonas sp.). Despite the morphological similarity of the red spores found at the snow surface, we found differences in intracellular organization investigated by light and transmission electron microscopy and in secondary pigments investigated by chromatographic analysis in combination with mass spectrometry. Spores of Chlainomonas sp. show clear differences from Chlamydomonas nivalis in cell wall arrangement and plastid organization. Active photosynthesis at ambient temperatures indicates a high physiological activity, despite no cell division being present. Lipid bodies containing the carotenoid astaxanthin, which produces the red color, dominate cells of both species, but are modified differently. While in Chlainomonas sp. astaxanthin is mainly esterified with two fatty acids and is more apolar, in Chamydomonas nivalis, in contrast, less apolar monoesters prevail. PMID:26884467

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

    PubMed Central

    Onishi, Masayuki; Pringle, John R.

    2016-01-01

    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. PMID:27770025

  19. Lead (Pb) and copper (Cu) share a common uptake transporter in the unicellular alga Chlamydomonas reinhardtii.

    PubMed

    Sánchez-Marín, Paula; Fortin, Claude; Campbell, Peter G C

    2014-02-01

    The unicellular alga Chlamydomonas reinhardtii has a very high rate of lead (Pb) internalization and is known to be highly sensitive to dissolved Pb. However, the transport pathway that this metal uses to cross cellular membranes in microalgae is still unknown. To identify the Pb(2+) transport pathway in C. reinhartdii, we performed several competition experiments with environmentally relevant concentrations of Pb(2+) (~10 nM) and a variety of divalent cations. Among the essential trace metals tested, cobalt, manganese, nickel and zinc had no effect on Pb internalization. A greater than tenfold increase in the concentrations of the major ions calcium and magnesium led to a slight decrease (~34 %) in short-term Pb internalization by the algae. Copper (Cu) was even more effective: at a Cu concentration 50 times higher than that of Pb, Pb internalization by the algae decreased by 87 %. Pre-exposure of the algae to Cu showed that the effect was not due to a physiological effect of Cu on the algae, but rather to competition for the same transporter. A reciprocal effect of Pb on Cu internalization was also observed. These results suggest that Cu and Pb share a common transport pathway in C. reinhardtii at environmentally relevant metal concentrations.

  20. Characterization of Chlamydomonas reinhardtii phosphatidylglycerophosphate synthase in Synechocystis sp. PCC 6803

    PubMed Central

    Hung, Chun-Hsien; Endo, Kaichiro; Kobayashi, Koichi; Nakamura, Yuki; Wada, Hajime

    2015-01-01

    Phosphatidylglycerol (PG) is an indispensable phospholipid class with photosynthetic function in plants and cyanobacteria. However, its biosynthesis in eukaryotic green microalgae is poorly studied. Here, we report the isolation and characterization of two homologs (CrPGP1 and CrPGP2) of phosphatidylglycerophosphate synthase (PGPS), the rate-limiting enzyme in PG biosynthesis, in Chlamydomonas reinhardtii. Heterologous complementation of Synechocystis sp. PCC 6803 pgsA mutant by CrPGP1 and CrPGP2 rescued the PG-dependent growth phenotype, but the PG level and its fatty acid composition were not fully rescued in the complemented strains. As well, oxygen evolution activity was not fully recovered, although electron transport activity of photosystem II was restored to the wild-type level. Gene expression study of CrPGP1 and CrPGP2 in nutrient-starved C. reinhardtii showed differential response to phosphorus and nitrogen deficiency. Taken together, these results highlight the distinct and overlapping function of PGPS in cyanobacteria and eukaryotic algae. PMID:26379630

  1. Genome analysis and its significance in four unicellular algae, Cyanidioschyzon [corrected] merolae, Ostreococcus tauri, Chlamydomonas reinhardtii, and Thalassiosira pseudonana.

    PubMed

    Misumi, Osami; Yoshida, Yamato; Nishida, Keiji; Fujiwara, Takayuki; Sakajiri, Takayuki; Hirooka, Syunsuke; Nishimura, Yoshiki; Kuroiwa, Tsuneyoshi

    2008-01-01

    Algae play a more important role than land plants in the maintenance of the global environment and productivity. Progress in genome analyses of these organisms means that we can now obtain information on algal genomes, global annotation and gene expression. The full genome information for several algae has already been analyzed. Whole genomes of the red alga Cyanidioschyzon [corrected] merolae, the green algae Ostreococcus tauri and Chlamydomonas reinhardtii, and the diatom Thalassiosira pseudonana have been sequenced. Genome composition and the features of cells among the four algae were compared. Each alga maintains basic genes as photosynthetic eukaryotes and possesses additional gene groups to represent their particular characteristics. This review discusses and introduces the latest research that makes the best use of the particular features of each organism and the significance of genome analysis to study biological phenomena. In particular, examples of post-genome studies of organelle multiplication in C. merolae based on analyzed genome information are presented.

  2. Carbon Supply and Photoacclimation Cross Talk in the Green Alga Chlamydomonas reinhardtii1[OPEN

    PubMed Central

    Fristedt, Rikard; Dinc, Emine

    2016-01-01

    Photosynthetic organisms are exposed to drastic changes in light conditions, which can affect their photosynthetic efficiency and induce photodamage. To face these changes, they have developed a series of acclimation mechanisms. In this work, we have studied the acclimation strategies of Chlamydomonas reinhardtii, a model green alga that can grow using various carbon sources and is thus an excellent system in which to study photosynthesis. Like other photosynthetic algae, it has evolved inducible mechanisms to adapt to conditions where carbon supply is limiting. We have analyzed how the carbon availability influences the composition and organization of the photosynthetic apparatus and the capacity of the cells to acclimate to different light conditions. Using electron microscopy, biochemical, and fluorescence measurements, we show that differences in CO2 availability not only have a strong effect on the induction of the carbon-concentrating mechanisms but also change the acclimation strategy of the cells to light. For example, while cells in limiting CO2 maintain a large antenna even in high light and switch on energy-dissipative mechanisms, cells in high CO2 reduce the amount of pigments per cell and the antenna size. Our results show the high plasticity of the photosynthetic apparatus of C. reinhardtii. This alga is able to use various photoacclimation strategies, and the choice of which to activate strongly depends on the carbon availability. PMID:27637747

  3. First crystal structure of Rubisco from a green alga, Chlamydomonas reinhardtii.

    PubMed

    Taylor, T C; Backlund, A; Bjorhall, K; Spreitzer, R J; Andersson, I

    2001-12-21

    The crystal structure of Rubisco (ribulose 1,5-bisphosphate carboxylase/oxygenase) from the unicellular green alga Chlamydomonas reinhardtii has been determined to 1.4 A resolution. Overall, the structure shows high similarity to the previously determined structures of L8S8 Rubisco enzymes. The largest difference is found in the loop between beta strands A and B of the small subunit (betaA-betaB loop), which is longer by six amino acid residues than the corresponding region in Rubisco from Spinacia. Mutations of residues in the betaA-betaB loop have been shown to affect holoenzyme stability and catalytic properties. The information contained in the Chlamydomonas structure enables a more reliable analysis of the effect of these mutations. No electron density was observed for the last 13 residues of the small subunit, which are assumed to be disordered in the crystal. Because of the high resolution of the data, some posttranslational modifications are unambiguously apparent in the structure. These include cysteine and N-terminal methylations and proline 4-hydroxylations.

  4. Lack of mutagenic activity of crude and refined oils in the unicellular alga Chlamydomonas reinhardtii

    SciTech Connect

    Vandermeulen, J.H.; Lee, R.W.

    1986-02-01

    Over the past several years, an increasing number of studies have presented evidence for the mutagenicity and/or carcinogenic potential of petroleum-derived hydrocarbons. These most usually were obtained with individual hydrocarbons, and using either specialized bacterial strains (e.g. Ames' strains) or mammalian tissue preparations. While providing important insights into mutagenic mechanisms involving xenobiotic compounds, the relevance of these studies to the natural aquatic environment is not always evident. This applies especially to the mutagenic potential of water-soluble fractions of hydrocarbon mixtures, as in whole oils or in complex distillate fractions, and involving typical marine biota. Accordingly, the authors have examined the mutagenic potential of the water-soluble fractions of four oils (two crude oils and two refined oils) using the unicellular haploid alga Chlamydomonas reinhardtii.

  5. Biochemical and morphological characterization of sulfur-deprived and H2-producing Chlamydomonas reinhardtii (green alga).

    PubMed

    Zhang, Liping; Happe, Thomas; Melis, Anastasios

    2002-02-01

    Sulfur deprivation in green algae causes reversible inhibition of photosynthetic activity. In the absence of S, rates of photosynthetic O2 evolution drop below those of O2 consumption by respiration. As a consequence, sealed cultures of the green alga Chlamydomonas reinhardtii become anaerobic in the light, induce the "Fe-hydrogenase" pathway of electron transport and photosynthetically produce H2 gas. In the course of such H2-gas production cells consume substantial amounts of internal starch and protein. Such catabolic reactions may sustain, directly or in directly, the H2-production process. Profile analysis of selected photosynthetic proteins showed a precipitous decline in the amount of ribulose-1,5-bisphosphate carboxylase-oxygenase (Rubisco) as a function of time in S deprivation, a more gradual decline in the level of photosystem (PS) II and PSI proteins, and a change in the composition of the PSII light-harvesting complex (LHC-II). An increase in the level of the enzyme Fe-hydrogenase was noted during the initial stages of S deprivation (0-72 h) followed by a decline in the level of this enzyme during longer (t >72 h) S-deprivation times. Microscopic observations showed distinct morphological changes in C. reinhardtii during S deprivation and H2 production. Ellipsoid-shaped cells (normal photosynthesis) gave way to larger and spherical cell shapes in the initial stages of S deprivation and H2 production, followed by cell mass reductions after longer S-deprivation and H2-production times. It is suggested that, under S-deprivation conditions, electrons derived from a residual PSII H2O-oxidation activity feed into the hydrogenase pathway, thereby contributing to the H2-production process in Chlamydomonas reinhardtii. Interplay between oxygenic photosynthesis, mitochondrial respiration, catabolism of endogenous substrate, and electron transport via the hydrogenase pathway is essential for this light-mediated H2-production process.

  6. Photosystem II stress tolerance in the unicellular green alga Chlamydomonas Reinhardtii under space conditions

    NASA Astrophysics Data System (ADS)

    Bertalan, Ivo; Esposito, Dania; Torzillo, Giuseppe; Faraloni, Cecilia; Johanningmeier, Udo; Giardi, Maria Teresa

    2007-09-01

    Photosynthesis was established on the earth 3.5 billion years ago. Due to the absence of the ozone layer in the early atmosphere it was most likely adapted to the presence of ionizing radiation continuously emitted by solar and stellar flares. That complex radiation spectrum comprises protons, alpha particles, heavy charged particle-HZE, electrons, X-ray and neutrons. Such spectrum has a significant impact on biological systems which capture light energy for e.g. photosynthesis. Oxygenic photosynthesis of plants, algae and cyanobacteria initiates at the level of photosystem II (PSII), a multisubunit protein complex embedded in the thylakoid membrane inside chloroplasts. PSII uses sunlight to power the unique photo-induced oxidation of water to atmospheric oxygen which is indispensable for most life forms. It is an especially sensitive component if exposed to space radiation and thus an important target for research aimed at improving bioregenerative life-support systems. The unicellular green algae Chlamydomonas reinhardtii is a long standing model organism for photosynthesis research. It was exposed to ionizing radiation in the ESA facility Biopan located in the Foton capsule brought to space by the Russian Soyuzfor 15 days. The algae were tested in space under shielded conditions in the past, but they were never exposed to direct ionizing radiation such as in Biopan. Conditions for survival were identified. It was observed that the effect of space stress on the survival of the algae varied depending on the light conditions to which they were exposed during the flight. In some cases the flight experience caused a stimulation of the photosystem II oxygen evolution of the cells.

  7. Controlling expression of genes in the unicellular alga Chlamydomonas reinhardtii with a vitamin-repressible riboswitch.

    PubMed

    Ramundo, Silvia; Rochaix, Jean-David

    2015-01-01

    Chloroplast genomes of land plants and algae contain generally between 100 and 150 genes. These genes are involved in plastid gene expression and photosynthesis and in various other tasks. The function of some chloroplast genes is still unknown and some of them appear to be essential for growth and survival. Repressible and reversible expression systems are highly desirable for functional and biochemical characterization of these genes. We have developed a genetic tool that allows one to regulate the expression of any coding sequence in the chloroplast genome of the unicellular alga Chlamydomonas reinhardtii. Our system is based on vitamin-regulated expression of the nucleus-encoded chloroplast Nac2 protein, which is specifically required for the expression of any plastid gene fused to the psbD 5'UTR. With this approach, expression of the Nac2 gene in the nucleus and, in turn, that of the chosen chloroplast gene artificially driven by the psbD 5'UTR, is controlled by the MetE promoter and Thi4 riboswitch, which can be inactivated in a reversible way by supplying vitamin B12 and thiamine to the growth medium, respectively. This system opens interesting possibilities for studying the assembly and turnover of chloroplast multiprotein complexes such as the photosystems, the ribosome, and the RNA polymerase. It also provides a way to overcome the toxicity often associated with the expression of proteins of biotechnological interest in the chloroplast.

  8. The Unicellular Green Alga Chlamydomonas reinhardtii as an Experimental System to Study Chloroplast RNA Metabolism

    NASA Astrophysics Data System (ADS)

    Nickelsen, J.; Kück, U.

    Chloroplasts are typical organelles of photoautotrophic eukaryotic cells which drive a variety of functions, including photosynthesis. For many years the unicellular green alga Chlamydomonas reinhardtii has served as an experimental organism for studying photosynthetic processes. The recent development of molecular tools for this organism together with efficient methods of genetic analysis and the availability of many photosynthesis mutants has now made this alga a powerful model system for the analysis of chloroplast biogenesis. For example, techniques have been developed to transfer recombinant DNA into both the nuclear and the chloroplast genome. This allows both complementation tests and analyses of gene functions in vivo. Moreover, site-specific DNA recombinations in the chloroplast allow targeted gene disruption experiments which enable a "reverse genetics" to be performed. The potential of the algal system for the study of chloroplast biogenesis is illustrated in this review by the description of regulatory systems of gene expression involved in organelle biogenesis. One example concerns the regulation of trans-splicing of chloroplast mRNAs, a process which is controlled by both multiple nuclear- and chloroplast-encoded factors. The second example involves the stabilization of chloroplast mRNAs. The available data lead us predict distinct RNA elements, which interact with trans-acting factors to protect the RNA against nucleolytic attacks.

  9. The Antarctic Psychrophile Chlamydomonas sp. UWO 241 Preferentially Phosphorylates a Photosystem I-Cytochrome b6/f Supercomplex.

    PubMed

    Szyszka-Mroz, Beth; Pittock, Paula; Ivanov, Alexander G; Lajoie, Gilles; Hüner, Norman P A

    2015-09-01

    Chlamydomonas sp. UWO 241 (UWO 241) is a psychrophilic green alga isolated from Antarctica. A unique characteristic of this algal strain is its inability to undergo state transitions coupled with the absence of photosystem II (PSII) light-harvesting complex protein phosphorylation. We show that UWO 241 preferentially phosphorylates specific polypeptides associated with an approximately 1,000-kD pigment-protein supercomplex that contains components of both photosystem I (PSI) and the cytochrome b₆/f (Cyt b₆/f) complex. Liquid chromatography nano-tandem mass spectrometry was used to identify three major phosphorylated proteins associated with this PSI-Cyt b₆/f supercomplex, two 17-kD PSII subunit P-like proteins and a 70-kD ATP-dependent zinc metalloprotease, FtsH. The PSII subunit P-like protein sequence exhibited 70.6% similarity to the authentic PSII subunit P protein associated with the oxygen-evolving complex of PSII in Chlamydomonas reinhardtii. Tyrosine-146 was identified as a unique phosphorylation site on the UWO 241 PSII subunit P-like polypeptide. Assessment of PSI cyclic electron transport by in vivo P700 photooxidation and the dark relaxation kinetics of P700(+) indicated that UWO 241 exhibited PSI cyclic electron transport rates that were 3 times faster and more sensitive to antimycin A than the mesophile control, Chlamydomonas raudensis SAG 49.72. The stability of the PSI-Cyt b₆/f supercomplex was dependent upon the phosphorylation status of the PsbP-like protein and the zinc metalloprotease FtsH as well as the presence of high salt. We suggest that adaptation of UWO 241 to its unique low-temperature and high-salt environment favors the phosphorylation of a PSI-Cyt b₆/f supercomplex to regulate PSI cyclic electron transport rather than the regulation of state transitions through the phosphorylation of PSII light-harvesting complex proteins.

  10. Toxicity assessment of manufactured nanomaterials using the unicellular green alga Chlamydomonas reinhardtii.

    PubMed

    Wang, Jiangxin; Zhang, Xuezhi; Chen, Yongsheng; Sommerfeld, Milton; Hu, Qiang

    2008-10-01

    With the rapid development of nanotechnology, there is an increasing risk of human and environmental exposure to nanotechnology-based materials and products. As water resources are particularly vulnerable to direct and indirect contamination of nonomaterials (NMs), the potential toxicity and environmental implication of NMs to aquatic organisms must be evaluated. In this study, we assessed potential toxicity of two commercially used NMs, titanium dioxide (TiO(2)) and quantum dots (QDs), using the unicellular green alga Chlamydomonas reinhartii as a model system. The response of the organism to NMs was assessed at physiological, biochemical, and molecular genetic levels. Growth kinetics showed that growth inhibition occurred during the first two to three days of cultivation in the presence of TiO(2) or QDs. Measurements of lipid peroxidation measurement indicated that oxidative stress of the cells occurred as early as 6 h after exposure to TiO(2) or QDs. The transcriptional expression profiling of four stress response genes (sod1, gpx, cat, and ptox2) revealed that transient up-regulation of these genes occurred in cultures containing as low as 1.0 mg L(-1) of TiO(2) or 0.1 mg L(-1) of QDs, and the maximum transcripts of cat, sod1, gpx, and ptox2 occurred at 1.5, 3, 3, and 6 h, respectively, and were proportional to the initial concentration of the NMs. As the cultures continued, recovery in growth was observed and the extent of recovery, as indicated by the final cell concentration, was dosage-dependent. QDs were found to be more toxic to Chlamydomonas cells than TiO(2) under our experimental conditions.

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

    SciTech Connect

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

    2014-10-18

    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. We demonstrate the growth of C. reinhardtii on acetate provided by mutant strains of the cyanobacterium Synechococcus sp. PCC7002.

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

    PubMed

    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.

  13. Transcriptome for Photobiological Hydrogen Production Induced by Sulfur Deprivation in the Green Alga Chlamydomonas reinhardtii▿ †

    PubMed Central

    Nguyen, Anh Vu; Thomas-Hall, Skye R.; Malnoë, Alizée; Timmins, Matthew; Mussgnug, Jan H.; Rupprecht, Jens; Kruse, Olaf; Hankamer, Ben; Schenk, Peer M.

    2008-01-01

    Photobiological hydrogen production using microalgae is being developed into a promising clean fuel stream for the future. In this study, microarray analyses were used to obtain global expression profiles of mRNA abundance in the green alga Chlamydomonas reinhardtii at different time points before the onset and during the course of sulfur-depleted hydrogen production. These studies were followed by real-time quantitative reverse transcription-PCR and protein analyses. The present work provides new insights into photosynthesis, sulfur acquisition strategies, and carbon metabolism-related gene expression during sulfur-induced hydrogen production. A general trend toward repression of transcripts encoding photosynthetic genes was observed. In contrast to all other LHCBM genes, the abundance of the LHCBM9 transcript (encoding a major light-harvesting polypeptide) and its protein was strongly elevated throughout the experiment. This suggests a major remodeling of the photosystem II light-harvesting complex as well as an important function of LHCBM9 under sulfur starvation and photobiological hydrogen production. This paper presents the first global transcriptional analysis of C. reinhardtii before, during, and after photobiological hydrogen production under sulfur deprivation. PMID:18708561

  14. Integration of carbon assimilation modes with photosynthetic light capture in the green alga Chlamydomonas reinhardtii.

    PubMed

    Berger, Hanna; Blifernez-Klassen, Olga; Ballottari, Matteo; Bassi, Roberto; Wobbe, Lutz; Kruse, Olaf

    2014-10-01

    The unicellular green alga Chlamydomonas reinhardtii is capable of using organic and inorganic carbon sources simultaneously, which requires the adjustment of photosynthetic activity to the prevailing mode of carbon assimilation. We obtained novel insights into the regulation of light-harvesting at photosystem II (PSII) following altered carbon source availability. In C. reinhardtii, synthesis of PSII-associated light-harvesting proteins (LHCBMs) is controlled by the cytosolic RNA-binding protein NAB1, which represses translation of particular LHCBM isoform transcripts. This mechanism is fine-tuned via regulation of the nuclear NAB1 promoter, which is activated when linear photosynthetic electron flow is restricted by CO(2)-limitation in a photoheterotrophic context. In the wild-type, accumulation of NAB1 reduces the functional PSII antenna size, thus preventing a harmful overexcited state of PSII, as observed in a NAB1-less mutant. We further demonstrate that translation control as a newly identified long-term response to prolonged CO(2)-limitation replaces LHCII state transitions as a fast response to PSII over-excitation. Intriguingly, activation of the long-term response is perturbed in state transition mutant stt7, suggesting a regulatory link between the long- and short-term response. We depict a regulatory circuit operating on distinct timescales and in different cellular compartments to fine-tune light-harvesting in photoheterotrophic eukaryotes.

  15. Effects of chromium on photosynthetic and photoreceptive apparatus of the alga Chlamydomonas reinhardtii.

    PubMed

    Rodríguez, M Cecilia; Barsanti, Laura; Passarelli, Vincenzo; Evangelista, Valter; Conforti, Visitacion; Gualtieri, Paolo

    2007-10-01

    Chromium is a highly toxic non-essential metal for microorganisms and plants. Due to its widespread industrial use, chromium (Cr) has become a serious pollutant in diverse environmental settings. The presence of Cr leads to the selection of algal populations able to tolerate high levels of Cr compounds. The diverse Cr-resistance mechanisms displayed by microorganisms include biosorption, diminished accumulation, precipitation, reduction of Cr(6+) to Cr(3+), and chromate efflux. In this paper we describe the effects of Cr(6+) (the more toxic species) on the photosynthetic and photoreceptive apparatus of the fresh water unicellular alga Chlamydomonas reinhardtii. We measured the effect of the heavy metal by means of in vivo absorption microspectroscopy of both the thylakoid compartments and the eyespot. The decomposition of the overall absorption spectra in pigment constituents indicates that Cr(6+) induced a complete pheophinitization of the chrorophylls and a modification of the carotenoids present in the eyespot only when its concentration is equal or greater than 10 microM. Due to this low tolerance level, C. reinhardtii could be used as indicator of Cr pollution, but it is not feasible for bioremediation purposes.

  16. Genome-wide characterization of genetic variation in the unicellular, green alga Chlamydomonas reinhardtii.

    PubMed

    Jang, Hyosik; Ehrenreich, Ian M

    2012-01-01

    Chlamydomonas reinhardtii is a model system for studying cilia, photosynthesis, and other core features of eukaryotes, and is also an emerging source of biofuels. Despite its importance to basic and applied biological research, the level and pattern of genetic variation in this haploid green alga has yet to be characterized on a genome-wide scale. To improve understanding of C. reinhardtii's genetic variability, we generated low coverage whole genome resequencing data for nearly all of the available isolates of this species, which were sampled from a number of sites in North America over the past ∼70 years. Based on the analysis of more than 62,000 single nucleotide polymorphisms, we identified two groups of isolates that represent geographical subpopulations of the species. We also found that measurements of genetic diversity were highly variable throughout the genome, in part due to technical factors. We studied the level and pattern of linkage disequilibrium (LD), and observed one chromosome that exhibits elevated LD. Furthermore, we detected widespread evidence of recombination across the genome, which implies that outcrossing occurs in natural populations of this species. In summary, our study provides multiple insights into the sequence diversity of C. reinhardtii that will be useful to future studies of natural genetic variation in this organism.

  17. Regulation of cellular manganese and manganese transport rates in the unicellular alga Chlamydomonas

    SciTech Connect

    Sunda, W.G.; Huntsman, S.A.

    1985-01-01

    The cellular accumulation and uptake kinetics of manganese by Chlamydomonas sp. were studied in model chelate buffer systems. Cellular manganese concentrations and uptake rates were related to the computed free manganese ion concentration and were independent of the total or chelated manganese concentration. Cellular manganese was constant at about 1 mmol liter/sup -1/ of cellular volume at free manganese ion concentrations of 10/sup -7/ /sup 6/-10/sup -6/ /sup 3/ mol liter/sup -1/ and decreased below this range. Manganese uptake rates followed saturation kinetics and V/sub max/, but not K/sub s/, varied with the free manganese ion concentration in the growth medium. V/sub max/ appeared to be under negative feedback control and increased with decreasing manganese ion concentration. Variations of up to 30-fold in this parameter seemed to be instrumental in limiting the variation in cellular manganese to a sixfold range despite a 1000-fold variation in free manganese ion concentration in the growth medium.

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

    PubMed

    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-H2DFFDA 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.

  19. Mastoparan-induced programmed cell death in the unicellular alga Chlamydomonas reinhardtii

    PubMed Central

    Yordanova, Zhenya P.; Woltering, Ernst J.; Kapchina-Toteva, Veneta M.; Iakimova, Elena T.

    2013-01-01

    Background and Aims Under stress-promoting conditions unicellular algae can undergo programmed cell death (PCD) but the mechanisms of algal cellular suicide are still poorly understood. In this work, the involvement of caspase-like proteases, DNA cleavage and the morphological occurrence of cell death in wasp venom mastoparan (MP)-treated Chlamydomonas reinhardtii were studied. Methods Algal cells were exposed to MP and cell death was analysed over time. Specific caspase inhibitors were employed to elucidate the possible role of caspase-like proteases. YVADase activity (presumably a vacuolar processing enzyme) was assayed by using a fluorogenic caspase-1 substrate. DNA breakdown was evaluated by DNA laddering and Comet analysis. Cellular morphology was examined by confocal laser scanning microscopy. Key Results MP-treated C. reinhardtii cells expressed several features of necrosis (protoplast shrinkage) and vacuolar cell death (lytic vesicles, vacuolization, empty cell-walled corpse-containing remains of digested protoplast) sometimes within one single cell and in different individual cells. Nucleus compaction and DNA fragmentation were detected. YVADase activity was rapidly stimulated in response to MP but the early cell death was not inhibited by caspase inhibitors. At later time points, however, the caspase inhibitors were effective in cell-death suppression. Conditioned medium from MP-treated cells offered protection against MP-induced cell death. Conclusions In C. reinhardtii MP triggered PCD of atypical phenotype comprising features of vacuolar and necrotic cell deaths, reminiscent of the modality of hypersensitive response. It was assumed that depending on the physiological state and sensitivity of the cells to MP, the early cell-death phase might be not mediated by caspase-like enzymes, whereas later cell death may involve caspase-like-dependent proteolysis. The findings substantiate the hypothesis that, depending on the mode of induction and sensitivity of

  20. Transcriptional and cellular responses of the green alga Chlamydomonas reinhardtii to perfluoroalkyl phosphonic acids.

    PubMed

    Sanchez, David; Houde, Magali; Douville, Mélanie; De Silva, Amila O; Spencer, Christine; Verreault, Jonathan

    2015-03-01

    Perfluoroalkyl phosphonic acids (PFPAs), a new class of perfluoroalkyl substances used primarily in the industrial sector as surfactants, were recently detected in surface water and wastewater treatment plant effluents. Toxicological effects of PFPAs have as yet not been investigated in aquatic organisms. The objective of the present study was to evaluate the effects of perfluorooctylphosphonic acid (C8-PFPA) and perfluorodecylphosphonic acid (C10-PFPA) exposure (31-250μg/L) on Chlamydomonas reinhardtii using genomic (qRT-PCR), biochemical (reactive oxygen species production (ROS) and lipid peroxidation), and physiological (cellular viability) indicators. After 72h of exposure, no differences were observed in cellular viability for any of the two perfluorochemicals. However, increase in ROS concentrations (36% and 25.6% at 125 and 250μg/L, respectively) and lipid peroxidation (35.5% and 35.7% at 125 and 250μg/L, respectively) was observed following exposure to C10-PFPA. C8-PFPA exposure did not impact ROS production and lipid peroxidation in algae. To get insights into the molecular response and modes of action of PFPA toxicity, qRT-PCR-based assays were performed to analyze the transcription of genes related to antioxidant responses including superoxide dismutase (SOD-1), glutathione peroxidase (GPX), catalase (CAT), glutathione S-transferase (GST), and ascorbate peroxidase (APX I). Genomic analyses revealed that the transcription of CAT and APX I was up-regulated for all the C10-PFPA concentrations. In addition, PFPAs were quantified in St. Lawrence River surface water samples and detected at concentrations ranging from 250 to 850pg/L for C8-PFPA and 380 to 650pg/L for C10-PFPA. This study supports the prevalence of PFPAs in the aquatic environment and suggests potential impacts of PFPA exposure on the antioxidant defensive system in C. reinhardtii.

  1. Cloning and expression analysis of two different LhcSR genes involved in stress adaptation in an Antarctic microalga, Chlamydomonas sp. ICE-L.

    PubMed

    Mou, Shanli; Zhang, Xiaowen; Ye, Naihao; Dong, Meitao; Liang, Chengwei; Liang, Qiang; Miao, Jinlai; Xu, Dong; Zheng, Zhou

    2012-03-01

    Light-harvesting complexes (LHCs) play essential roles in light capture and photoprotection. Although the functional diversity of individual LHCs in many plants has been well described, knowledge regarding the extent of this family in the majority of green algal groups is still limited. In this study, two different LhcSR genes, LhcSR1 and LhcSR2 from Chlamydomonas sp. ICE-L, were cloned from the total cDNA and characterized in response to high light (HL), low light (LL), UV-B radiation and high salinity. The lower F (v)/F (m) as well as the associated induction of non-photochemical quenching (NPQ), observed under those conditions, indicated that Chlamydomonas sp. ICE-L was under stress. Under HL stress, the expression of LhcSR1 and LhcSR2 increased rapidly from 0.5 h HL and reached a maximum after 3 h. In LL, LhcSR2 expression was up-regulated during the first 0.5 h after which it decreased, while the expression of LhcSR1 decreased gradually from the beginning of the experiment. In addition, the transcript levels of LhcSR1 and LhcSR2 increased under UV-B radiation and high salinity. These results showed that both genes were inducible and up-regulated under stress conditions. A higher NPQ was accompanied by the up-regulated LhcSR genes, suggesting that LhcSR plays a role in thermal energy dissipation. Overall, the results presented here suggest that LhcSR1 and LhcSR2 play a primary role in photoprotection in Chlamydomonas sp. ICE-L under stress conditions and provide an important basis for investigation of the adaptation mechanism of LhcSR in Antarctic green algae.

  2. Antioxidant capacity, polyphenol content and iron bioavailability from algae (Ulva sp., Sargassum sp. and Porphyra sp.) in human subjects.

    PubMed

    García-Casal, Maria N; Ramírez, José; Leets, Irene; Pereira, Ana C; Quiroga, Maria F

    2009-01-01

    Marine algae are easily produced and are good sources of Fe. If this Fe is bioavailable, algae consumption could help to combat Fe deficiency and anaemia worldwide. The objective of the present study was to evaluate Fe bioavailability, polyphenol content and antioxidant capacity from three species of marine algae distributed worldwide. A total of eighty-three subjects received maize- or wheat-based meals containing marine algae (Ulva sp., Sargassum sp. and Porphyra sp.) in different proportions (2.5, 5.0 and 7.5 g) added to the water to prepare the dough. All meals administered contained radioactive Fe. Absorption was evaluated calculating radioactive Fe incorporation in subjects' blood. The three species of marine algae were analysed for polyphenol content and reducing power. Algae significantly increased Fe absorption in maize- or wheat-based meals, especially Sargassum sp., due to its high Fe content. Increases in absorption were dose-dependent and higher in wheat- than in maize-based meals. Total polyphenol content was 10.84, 18.43 and 80.39 gallic acid equivalents/g for Ulva sp., Porphyra sp. and Sargassum sp., respectively. The antioxidant capacity was also significantly higher in Sargassum sp. compared with the other two species analysed. Ulva sp., Sargassum sp. and Porphyra sp. are good sources of bioavailable Fe. Sargassum sp. resulted in the highest Fe intake due to its high Fe content, and a bread containing 7.5 g Sargassum sp. covers daily Fe needs. The high polyphenol content found in Sargassum sp. could be partly responsible for the antioxidant power reported here, and apparently did not affect Fe absorption.

  3. Refactoring the Six-Gene Photosystem II Core in the Chloroplast of the Green Algae Chlamydomonas reinhardtii.

    PubMed

    Gimpel, Javier A; Nour-Eldin, Hussam H; Scranton, Melissa A; Li, Daphne; Mayfield, Stephen P

    2016-07-15

    Oxygenic photosynthesis provides the energy to produce all food and most of the fuel on this planet. Photosystem II (PSII) is an essential and rate-limiting component of this process. Understanding and modifying PSII function could provide an opportunity for optimizing photosynthetic biomass 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. Complementation of the knockout strain with the core PSII synthetic module from three different green algae resulted in reconstitution of photosynthetic activity to 85, 55, and 53% of that of the wild-type, demonstrating that the PSII core can be exchanged between algae species and retain function. The strains, synthetic cassettes, and refactoring strategy developed for this study demonstrate the potential of synthetic biology approaches for tailoring oxygenic photosynthesis and provide a powerful tool for unraveling PSII structure-function relationships.

  4. Deep Transcriptome Sequencing of Two Green Algae, Chara vulgaris and Chlamydomonas reinhardtii, Provides No Evidence of Organellar RNA Editing

    PubMed Central

    Cahoon, A. Bruce; Nauss, John A.; Stanley, Conner D.; Qureshi, Ali

    2017-01-01

    Nearly all land plants post-transcriptionally modify specific nucleotides within RNAs, a process known as RNA editing. This adaptation allows the correction of deleterious mutations within the asexually reproducing and presumably non-recombinant chloroplast and mitochondrial genomes. There are no reports of RNA editing in any of the green algae so this phenomenon is presumed to have originated in embryophytes either after the invasion of land or in the now extinct algal ancestor of all land plants. This was challenged when a recent in silico screen for RNA edit sites based on genomic sequence homology predicted edit sites in the green alga Chara vulgaris, a multicellular alga found within the Streptophyta clade and one of the closest extant algal relatives of land plants. In this study, the organelle transcriptomes of C. vulgaris and Chlamydomonas reinhardtii were deep sequenced for a comprehensive assessment of RNA editing. Initial analyses based solely on sequence comparisons suggested potential edit sites in both species, but subsequent high-resolution melt analysis, RNase H-dependent PCR (rhPCR), and Sanger sequencing of DNA and complementary DNAs (cDNAs) from each of the putative edit sites revealed them to be either single-nucleotide polymorphisms (SNPs) or spurious deep sequencing results. The lack of RNA editing in these two lineages is consistent with the current hypothesis that RNA editing evolved after embryophytes split from its ancestral algal lineage. PMID:28230734

  5. The mTERF protein MOC1 terminates mitochondrial DNA transcription in the unicellular green alga Chlamydomonas reinhardtii.

    PubMed

    Wobbe, Lutz; Nixon, Peter J

    2013-07-01

    The molecular function of mTERFs (mitochondrial transcription termination factors) has so far only been described for metazoan members of the protein family and in animals they control mitochondrial replication, transcription and translation. Cells of photosynthetic eukaryotes harbour chloroplasts and mitochondria, which are in an intense cross-talk that is vital for photosynthesis. Chlamydomonas reinhardtii is a unicellular green alga widely used as a model organism for photosynthesis research and green biotechnology. Among the six nuclear C. reinhardtii mTERF genes is mTERF-like gene of Chlamydomonas (MOC1), whose inactivation alters mitorespiration and interestingly also light-acclimation processes in the chloroplast that favour the enhanced production of biohydrogen. We show here from in vitro studies that MOC1 binds specifically to a sequence within the mitochondrial rRNA-coding module S3, and that a knockout of MOC1 in the mutant stm6 increases read-through transcription at this site, indicating that MOC1 acts as a transcription terminator in vivo. Whereas the level of certain antisense RNA species is higher in stm6, the amount of unprocessed mitochondrial sense transcripts is strongly reduced, demonstrating that a loss of MOC1 causes perturbed mitochondrial DNA (mtDNA) expression. Overall, we provide evidence for the existence of mitochondrial antisense RNAs in C. reinhardtii and show that mTERF-mediated transcription termination is an evolutionary-conserved mechanism occurring in phototrophic protists and metazoans.

  6. The mTERF protein MOC1 terminates mitochondrial DNA transcription in the unicellular green alga Chlamydomonas reinhardtii

    PubMed Central

    Wobbe, Lutz; Nixon, Peter J.

    2013-01-01

    The molecular function of mTERFs (mitochondrial transcription termination factors) has so far only been described for metazoan members of the protein family and in animals they control mitochondrial replication, transcription and translation. Cells of photosynthetic eukaryotes harbour chloroplasts and mitochondria, which are in an intense cross-talk that is vital for photosynthesis. Chlamydomonas reinhardtii is a unicellular green alga widely used as a model organism for photosynthesis research and green biotechnology. Among the six nuclear C. reinhardtii mTERF genes is mTERF-like gene of Chlamydomonas (MOC1), whose inactivation alters mitorespiration and interestingly also light-acclimation processes in the chloroplast that favour the enhanced production of biohydrogen. We show here from in vitro studies that MOC1 binds specifically to a sequence within the mitochondrial rRNA-coding module S3, and that a knockout of MOC1 in the mutant stm6 increases read-through transcription at this site, indicating that MOC1 acts as a transcription terminator in vivo. Whereas the level of certain antisense RNA species is higher in stm6, the amount of unprocessed mitochondrial sense transcripts is strongly reduced, demonstrating that a loss of MOC1 causes perturbed mitochondrial DNA (mtDNA) expression. Overall, we provide evidence for the existence of mitochondrial antisense RNAs in C. reinhardtii and show that mTERF-mediated transcription termination is an evolutionary-conserved mechanism occurring in phototrophic protists and metazoans. PMID:23649833

  7. Cooperative processing of primary miRNAs by DUS16 and DCL3 in the unicellular green alga Chlamydomonas reinhardtii

    PubMed Central

    Cerutti, Heriberto

    2017-01-01

    ABSTRACT We have previously reported that the RNA-binding protein Dull slicer 16 (DUS16) plays a key role in the processing of primary miRNAs (pri-miRNAs) in the unicellular green alga Chlamydomonas reinhardtii. In the present report, we elaborate on the interaction of DUS16 with Dicer-like 3 (DCL3) during pri-miRNA processing. Comprehensive analyses of small RNA libraries derived from mutant and wild-type algal strains allowed the de novo prediction of 35 pri-miRNA genes, including 9 previously unknown ones. The pri-miRNAs dependent on DUS16 for processing largely overlapped with those dependent on DCL3. Our findings suggest that DUS16 and DCL3 work cooperatively, presumably as components of a microprocessor complex, in the processing of the majority of pri-miRNAs in C. reinhardtii. PMID:28289490

  8. The ferredoxin-thioredoxin system of a green alga, Chlamydomonas reinhardtii: identification and characterization of thioredoxins and ferredoxin-thioredoxin reductase components

    NASA Technical Reports Server (NTRS)

    Huppe, H. C.; de Lamotte-Guery, F.; Buchanan, B. B.

    1990-01-01

    The components of the ferredoxin-thioredoxin (FT) system of Chlamydomonas reinhardtii have been purified and characterized. The system resembled that of higher plants in consisting of a ferredoxin-thioredoxin reductase (FTR) and two types of thioredoxin, a single f and two m species, m1 and m2. The Chlamydomonas m and f thioredoxins were antigenically similar to their higher-plant counterparts, but not to one another. The m thioredoxins were recognized by antibodies to both higher plant m and bacterial thioredoxins, whereas the thioredoxin f was not. Chlamydomonas thioredoxin f reacted, although weakly, with the antibody to spinach thioredoxin f. The algal thioredoxin f differed from thioredoxins studied previously in behaving as a basic protein on ion-exchange columns. Purification revealed that the algal thioredoxins had molecular masses (Mrs) typical of thioredoxins from other sources, m1 and m2 being 10700 and f 11500. Chlamydomonas FTR had two dissimilar subunits, a feature common to all FTRs studied thus far. One, the 13-kDa ("similar") subunit, resembled its counterpart from other sources in both size and antigenicity. The other, 10-kDa ("variable") subunit was not recognized by antibodies to any FTR tested. When combined with spinach, (Spinacia oleracea L.) thylakoid membranes, the components of the FT system functioned in the light activation of the standard target enzymes from chloroplasts, corn (Zea mays L.) NADP-malate dehydrogenase (EC 1.1.1.82) and spinach fructose 1,6-bisphosphatase (EC 3.1.3.11) as well as the chloroplast-type fructose 1,6-bisphosphatase from Chlamydomonas. Activity was greatest if ferredoxin and other components of the FT system were from Chlamydomonas. The capacity of the Chlamydomonas FT system to activate autologous FBPase indicates that light regulates the photosynthetic carbon metabolism of green algae as in other oxygenic photosynthetic organisms.

  9. Survival and proliferation characteristics of the microalga Chlamydomonas sp. ICE-L after hypergravitational stress pretreatment

    NASA Astrophysics Data System (ADS)

    Gao, Zhengquan; Li, Demao; Meng, Chunxiao; Xu, Dong; Zhang, Xiaowen; Ye, Naihao

    2013-09-01

    Seeking extraterrestrial life, transferring between planets, even migrating to other planets attracts more and more attention of public and scientists. However, to make it clear for the ability to survive the forces studies is prerequisite to enable the speculations by natural means. Gravity is a critical force involved in all the life on Earth and, possibly, others planets. Organisms have been grown in microgravity habitats and in centrifuges to characterize the biological response to a range of gravitational forces and radiation levels in space and on Earth. However, little is known about the profiles of eukaryotic life under conditions of hyperacceleration attributable to extreme gravities. In this study, a eukaryotic extremophile, the Antarctic green microalga Chlamydomonas sp. ICE-L, showed amazing proliferation capacity during and after hypergravitational stress for 30 min to 48 h at 110,200, 423,400, and 670,800g. These extreme gravities also had profound effects on viability, reproduction rate, photosynthesis efficiency, and gene transcriptional expression of this microalga. Most notably, all three supergravities efficiently stimulated algal cell division, but the greater the centrifugal force and the longer the duration of treatment, the lower the viable rate and breeding potential of samples in the following incubation. These results illustrated Chlamydomonas sp. ICE-L is a useful eukaryotic model system candidate for space research. Further studies could provide new insight into the physical limits of life and its evolution and enhance the possibility for interstellar space travel and the quest for extraterrestrial life according to panspermia theory. Also, it indicated that life come from the outer space is not always prokaryotes but may be eukaryotes.

  10. Cultivation of Monoraphidium sp., Chlorella sp. and Scenedesmus sp. algae in Batch culture using Nile tilapia effluent.

    PubMed

    Guerrero-Cabrera, Luis; Rueda, José A; García-Lozano, Hiram; Navarro, A Karin

    2014-06-01

    Monoraphidium sp., Chlorella sp. and Scenedesmus sp. algae were cultured in three volumes of Tilapia Effluent Medium (TEM) in comparison with the Bold Basal Medium (BBM) (Nichols and Bold, 1965). Specific growth rate (μ'), biomass dry productivity (Q), volumetric productivity (Qv) as well as lipid and protein content were measured. Then, volumetric productivities for both lipids and proteins were calculated (QVL and QVP). In Scenedesmus sp., BBM produced higher μ' and Qv than TEM in 1.5L volume. Chlorella sp. showed a higher QVL for BBM than TEM. Any observed difference in protein or lipid productivities among volumes was in favor of a greater productivity for 1.5L volume. Even when TEM had a larger protein content in Chlorella sp. than BBM, QVP was not different. Current results imply that TEM can be used as an alternative growth medium for algae when using Batch cultures, yet productivity is reduced.

  11. Rubisco small subunits from the unicellular green alga Chlamydomonas complement Rubisco-deficient mutants of Arabidopsis.

    PubMed

    Atkinson, Nicky; Leitão, Nuno; Orr, Douglas J; Meyer, Moritz T; Carmo-Silva, Elizabete; Griffiths, Howard; Smith, Alison M; McCormick, Alistair J

    2017-04-01

    Introducing components of algal carbon concentrating mechanisms (CCMs) into higher plant chloroplasts could increase photosynthetic productivity. A key component is the Rubisco-containing pyrenoid that is needed to minimise CO2 retro-diffusion for CCM operating efficiency. Rubisco in Arabidopsis was re-engineered to incorporate sequence elements that are thought to be essential for recruitment of Rubisco to the pyrenoid, namely the algal Rubisco small subunit (SSU, encoded by rbcS) or only the surface-exposed algal SSU α-helices. Leaves of Arabidopsis rbcs mutants expressing 'pyrenoid-competent' chimeric Arabidopsis SSUs containing the SSU α-helices from Chlamydomonas reinhardtii can form hybrid Rubisco complexes with catalytic properties similar to those of native Rubisco, suggesting that the α-helices are catalytically neutral. The growth and photosynthetic performance of complemented Arabidopsis rbcs mutants producing near wild-type levels of the hybrid Rubisco were similar to those of wild-type controls. Arabidopsis rbcs mutants expressing a Chlamydomonas SSU differed from wild-type plants with respect to Rubisco catalysis, photosynthesis and growth. This confirms a role for the SSU in influencing Rubisco catalytic properties.

  12. [Effect of Methylmercury on the Light Dependence Fluorescence Parameters in a Green Alga Chlamydomonas moewusii].

    PubMed

    Protopopov, F F; Matorin, D N; Seifullina, N H; Bratkovskaya, L B; Zayadan, B K

    2015-01-01

    The effect of a dangerous toxic substance, methylmercury, on light dependence curves of chlorophyll fluorescence in Chlamydomonas moewusii was studied. We found low concentration of methylmercury (10(-7) M) to cause a decrease in the relative rate of the non-cyclic electron transport activity of PS 2, a decline in the maximum utilization of light energy (α), and a decline in the saturation light intensity (E(s)). Non-photochemical fluorescence quenching increased after short-term exposure and decreased in the course of prolonged incubation. These parameters were more sensitive to the action of the toxic substance than the widely used parameter F(V)/F(M), which reflects the maximum quantum yield of PS 2. We propose the use of the method of fast measurement of light dependence curves of fluorescence to detect the changes in algal cells at the early stages of exposure to mercury salts.

  13. Enzymatic modification by point mutation and functional analysis of an omega-6 fatty acid desaturase from Arctic Chlamydomonas sp.

    PubMed

    Jung, Woongsic; Kim, Eun Jae; Han, Se Jong; Kang, Sung-Ho; Choi, Han-Gu; Kim, Sanghee

    2017-02-07

    Arctic Chlamydomonas sp. is a dominant microalgal strain in cold or frozen freshwater in the Arctic region. The full-length open reading frame of the omega-6 fatty acid desaturase gene (AChFAD6) was obtained from the transcriptomic database of Arctic Chlamydomonas sp. from the KOPRI culture collection of polar micro-organisms. Amino acid sequence analysis indicated the presence of three conserved histidine-rich segments as unique characteristics of omega-6 fatty acid desaturases, and three transmembrane regions transported to plastidic membranes by chloroplast transit peptides in the N-terminal region. The AChFAD6 desaturase activity was examined by expressing wild-type and V254A mutant (Mut-AChFAD6) heterologous recombinant proteins. Quantitative gas chromatography indicated that the concentration of linoleic acids in AChFAD6-transformed cells increased more than 3-fold [6.73 ± 0.13 mg g(-1) dry cell weight (DCW)] compared with cells transformed with vector alone. In contrast, transformation with Mut-AChFAD6 increased the concentration of oleic acid to 9.23 ± 0.18 mg g(-1) DCW, indicating a change in enzymatic activity to mimic that of stearoyl-CoA desaturase. These results demonstrate that AChFAD6 of Arctic Chlamydomonas sp. increases membrane fluidity by enhancing denaturation of C18 fatty acids and facilitates production of large quantities of linoleic fatty acids in prokaryotic expression systems.

  14. Involvement of ethylene and nitric oxide in cell death in mastoparan-treated unicellular alga Chlamydomonas reinhardtii.

    PubMed

    Yordanova, Zhenya P; Iakimova, Elena T; Cristescu, Simona M; Harren, Frans J M; Kapchina-Toteva, Veneta M; Woltering, Ernst J

    2010-02-22

    This work demonstrates a contribution of ethylene and NO (nitric oxide) in MP (mastoparan)-induced cell death in the green algae Chlamydomonas reinhardtii. Following MP treatment, C. reinhardtii showed massive cell death, expressing morphological features of PCD (programmed cell death). A pharmacological approach involving combined treatments with MP and ethylene- and NO-interacting compounds indicated the requirement of trace amounts of both ethylene and NO in MP-induced cell death. By employing a carbon dioxide laser-based photoacoustic detector to measure ethylene and a QCL (quantum cascade laser)-based spectrometer for NO detection, simultaneous increases in the production of both ethylene and NO were observed following MP application. Our results show a tight regulation of the levels of both signalling molecules in which ethylene stimulates NO production and NO stimulates ethylene production. This suggests that, in conjunction with the elicitor, NO and ethylene cooperate and act synchronously in the mediation of MP-induced PCD in C. reinhardtii. To the best of our knowledge, this is the first report on the functional significance of ethylene and NO in MP-induced cell death.

  15. Origin of pronounced differences in 77 K fluorescence of the green alga Chlamydomonas reinhardtii in state 1 and 2.

    PubMed

    Ünlü, Caner; Polukhina, Iryna; van Amerongen, Herbert

    2016-04-01

    In response to changes in the reduction state of the plastoquinone pool in its thylakoid membrane, the green alga Chlamydomonas reinhardtti is performing state transitions: remodelling of its thylakoid membrane leads to a redistribution of excitations over photosystems I and II (PSI and PSII). These transitions are accompanied by marked changes in the 77 K fluorescence spectrum, which form the accepted signature of state transitions. The changes are generally thought to reflect a redistribution of light-harvesting complexes (LHCs) over PSII (fluorescing below 700 nm) and PSI (fluorescing above 700 nm). Here we studied the picosecond fluorescence properties of C. reinhardtti over a broad range of wavelengths with very low excitation intensities (0.2 nJ per laser pulse). Cells were directly used for time-resolved fluorescence measurements at 77 K without further treatment, such as medium exchange with glycerol. It is observed that upon going from state 1 (relatively more fluorescence below 700 nm) to state 2 (relatively more fluorescence above 700 nm), a large part of the fluorescence of LHC/PSII becomes substantially quenched in concurrence with LHC detachment from PSII, whereas the absolute amount of PSI fluorescence hardly changes. These results are in agreement with the recent proposal that the amount of LHC moving from PSII to PSI upon going from state 1 to state 2 is rather limited (Unlu et al. Proc Natl Acad Sci USA 111 (9):3460-3465, 2014).

  16. Effect of petroleum hydrocarbons on algae

    SciTech Connect

    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.

  17. Comparison of the Structural Changes Occurring during the Primary Phototransition of Two Different Channelrhodopsins from Chlamydomonas Algae

    PubMed Central

    2015-01-01

    Channelrhodopsins (ChRs) from green flagellate algae function as light-gated ion channels when expressed heterologously in mammalian cells. Considerable interest has focused on understanding the molecular mechanisms of ChRs to bioengineer their properties for specific optogenetic applications such as elucidating the function of specific neurons in brain circuits. While most studies have used channelrhodopsin-2 from Chlamydomonas reinhardtii (CrChR2), in this work low-temperature Fourier transform infrared-difference spectroscopy is applied to study the conformational changes occurring during the primary phototransition of the red-shifted ChR1 from Chlamydomonas augustae (CaChR1). Substitution with isotope-labeled retinals or the retinal analogue A2, site-directed mutagenesis, hydrogen–deuterium exchange, and H218O exchange were used to assign bands to the retinal chromophore, protein, and internal water molecules. The primary phototransition of CaChR1 at 80 K involves, in contrast to that of CrChR2, almost exclusively an all-trans to 13-cis isomerization of the retinal chromophore, as in the primary phototransition of bacteriorhodopsin (BR). In addition, significant differences are found for structural changes of the protein and internal water(s) compared to those of CrChR2, including the response of several Asp/Glu residues to retinal isomerization. A negative amide II band is identified in the retinal ethylenic stretch region of CaChR1, which reflects along with amide I bands alterations in protein backbone structure early in the photocycle. A decrease in the hydrogen bond strength of a weakly hydrogen bonded internal water is detected in both CaChR1 and CrChR2, but the bands are much broader in CrChR2, indicating a more heterogeneous environment. Mutations involving residues Glu169 and Asp299 (homologues of the Asp85 and Asp212 Schiff base counterions, respectively, in BR) lead to the conclusion that Asp299 is protonated during P1 formation and suggest that

  18. Characterization of the Major Light-Harvesting Complexes (LHCBM) of the Green Alga Chlamydomonas reinhardtii

    PubMed Central

    Natali, Alberto; Croce, Roberta

    2015-01-01

    Nine genes (LHCBM1-9) encode the major light-harvesting system of Chlamydomonas reinhardtii. Transcriptomic and proteomic analyses have shown that those genes are all expressed albeit in different amounts and some of them only in certain conditions. However, little is known about the properties and specific functions of the individual gene products because they have never been isolated. Here we have purified several complexes from native membranes and/or we have reconstituted them in vitro with pigments extracted from C. reinhardtii. It is shown that LHCBM1 and -M2/7 represent more than half of the LHCBM population in the membrane. LHCBM2/7 forms homotrimers while LHCBM1 seems to be present in heterotrimers. Trimers containing only type I LHCBM (M3/4/6/8/9) were also observed. Despite their different roles, all complexes have very similar properties in terms of pigment content, organization, stability, absorption, fluorescence and excited-state lifetimes. Thus the involvement of LHCBM1 in non-photochemical quenching is suggested to be due to specific interactions with other components of the membrane and not to the inherent quenching properties of the complex. Similarly, the overexpression of LHCBM9 during sulfur deprivation can be explained by its low sulfur content as compared with the other LHCBMs. Considering the highly conserved biochemical and spectroscopic properties, the major difference between the complexes may be in their capacity to interact with other components of the thylakoid membrane. PMID:25723534

  19. Spontaneous mutation accumulation in multiple strains of the green alga, Chlamydomonas reinhardtii.

    PubMed

    Morgan, Andrew D; Ness, Rob W; Keightley, Peter D; Colegrave, Nick

    2014-09-01

    Estimates of mutational parameters, such as the average fitness effect of a new mutation and the rate at which new genetic variation for fitness is created by mutation, are important for the understanding of many biological processes. However, the causes of interspecific variation in mutational parameters and the extent to which they vary within species remain largely unknown. We maintained multiple strains of the unicellular eukaryote Chlamydomonas reinhardtii, for approximately 1000 generations under relaxed selection by transferring a single cell every ~10 generations. Mean fitness of the lines tended to decline with generations of mutation accumulation whereas mutational variance increased. We did not find any evidence for differences among strains in any of the mutational parameters estimated. The overall change in mean fitness per cell division and rate of input of mutational variance per cell division were more similar to values observed in multicellular organisms than to those in other single-celled microbes. However, after taking into account differences in genome size among species, estimates from multicellular organisms and microbes, including our new estimates from C. reinhardtii, become substantially more similar. Thus, we suggest that variation in genome size is an important determinant of interspecific variation in mutational parameters.

  20. Towards elucidation of the toxic mechanism of copper on the model green alga Chlamydomonas reinhardtii.

    PubMed

    Jiang, Yongguang; Zhu, Yanli; Hu, Zhangli; Lei, Anping; Wang, Jiangxin

    2016-09-01

    Toxic effects of copper on aquatic organisms in polluted water bodies have garnered particular attention in recent years. Microalgae play an important role in aquatic ecosystems, and they are sensitive to heavy metal pollution. Thus, it is important to clarify the mechanism of copper toxicity first for ecotoxicology studies. In this study, the physiological, biochemical and gene expression characteristics of a model green microalga, Chlamydomonas reinhardtii, with 0, 50, 150 and 250 μM copper treatments were investigated. The response of C. reinhardtii to copper stress was significantly shown at a dose dependent manner. Inhibition of cell growth and variation of total chlorophyll content were observed with copper treatments. The maximum photochemical efficiency of PSII, actual photochemical efficiency of PSII and photochemical quenching value decreased in the 250 μM copper treatment with minimum values equal to 28, 24 and 60 % of the control values respectively. The content of lipid peroxidation biomarker malondialdehyde with copper treatments increased with a maximum value sevenfold higher than the control value. Inhibition of cell growth and photosynthesis was ascribed to peroxidation of membrane lipids. The glutathione content and activities of antioxidant enzymes, glutathione S-transferase, glutathione peroxidase, superoxide dismutase and peroxidase were induced by copper. Interestingly, the expression of antioxidant genes and the photosynthetic gene decreased in most copper treatments. In conclusion, oxidative stress caused by production of excess reactive oxidative species might be the major mechanism of copper toxicity on C. reinhardtii.

  1. Ecophysiology, secondary pigments and ultrastructure of Chlainomonas sp. (Chlorophyta) from the European Alps compared with Chlamydomonas nivalis forming red snow.

    PubMed

    Remias, Daniel; Pichrtová, Martina; Pangratz, Marion; Lütz, Cornelius; Holzinger, Andreas

    2016-04-01

    Red snow is a well-known phenomenon caused by microalgae thriving in alpine and polar regions during the melting season. The ecology and biodiversity of these organisms, which are adapted to low temperatures, high irradiance and freeze-thaw events, are still poorly understood. We compared two different snow habitats containing two different green algal genera in the European Alps, namely algae blooming in seasonal rock-based snowfields (Chlamydomonas nivalis) and algae dominating waterlogged snow bedded over ice (Chlainomonassp.). Despite the morphological similarity of the red spores found at the snow surface, we found differences in intracellular organization investigated by light and transmission electron microscopy and in secondary pigments investigated by chromatographic analysis in combination with mass spectrometry. Spores ofChlainomonassp. show clear differences fromChlamydomonas nivalisin cell wall arrangement and plastid organization. Active photosynthesis at ambient temperatures indicates a high physiological activity, despite no cell division being present. Lipid bodies containing the carotenoid astaxanthin, which produces the red color, dominate cells of both species, but are modified differently. While inChlainomonassp. astaxanthin is mainly esterified with two fatty acids and is more apolar, inChamydomonas nivalis, in contrast, less apolar monoesters prevail.

  2. Manipulating RuBisCO accumulation in the green alga, Chlamydomonas reinhardtii.

    PubMed

    Johnson, Xenie

    2011-07-01

    The nuclear factor, Maturation/stability of RbcL (MRL1), regulates the accumulation of the chloroplast rbcL gene transcript in Chlamydomonas reinhardtii by stabilising the mRNA via its 5' UTR. An absence of MRL1 in algal mrl1 mutants leads to a complete absence of RuBisCO large subunit protein and thus a lack of accumulation of the RuBisCO holoenzyme. By complementing mrl1 mutants by random transformation of the nuclear genome with the MRL1 cDNA, different levels of rbcL transcript accumulate. We also observe that RuBisCO Large Subunit accumulation is perturbed. Complemented strains accumulating as little as 15% RuBisCO protein can grow phototrophically while RuBisCO in this range is limiting for phototrophic growth. We also observe that photosynthetic activity, here measured by the quantum yield of PSII, appears to be a determinant for phototrophic growth. In some strains that accumulate less RuBisCO, a strong production of reactive oxygen species is detected. In the absence of RuBisCO, oxygen possibly acts as the PSI terminal electron acceptor. These results show that random transformation of MRL1 into mrl1 mutants can change RuBisCO accumulation allowing a range of phototrophic growth phenotypes. Furthermore, this technique allows for the isolation of strains with low RuBisCO, within the range of acceptable photosynthetic growth and reasonably low ROS production. MRL1 is thus a potential tool for applications to divert electrons away from photosynthetic carbon metabolism towards alternative pathways.

  3. Two Loci Control Phytoglycogen Production in the Monocellular Green Alga Chlamydomonas reinhardtii1

    PubMed Central

    Dauvillée, David; Colleoni, Christophe; Mouille, Gregory; Buléon, Alain; Gallant, Daniel J.; Bouchet, Brigitte; Morell, Matthew K.; d'Hulst, Christophe; Myers, Alan M.; Ball, Steven G.

    2001-01-01

    The STA8 locus of Chlamydomonas reinhardtii was identified in a genetic screen as a factor that controls starch biosynthesis. Mutations of STA8 cause a significant reduction in the amount of granular starch produced during nutrient limitation and accumulate phytoglycogen. The granules remaining in sta8 mutants are misshapen, and the abundance of amylose and long chains in amylopectin is altered. Mutations of the STA7 locus, which completely lack isoamylase activity, also cause accumulation of phytoglycogen, although sta8 and sta7 mutants differ in that there is a complete loss of granular starch in the latter. This is the first instance in which mutations of two different genetic elements in one plant species have been shown to cause phytoglycogen accumulation. An analytical procedure that allows assay of isoamylase in total extracts was developed and used to show that sta8 mutations cause a 65% reduction in the level of this activity. All other enzymes known to be involved in starch biosynthesis were shown to be unaffected in sta8 mutants. The same amount of total isoamylase activity (approximately) as that present in sta8 mutants was observed in heterozygous triploids containing two sta7 mutant alleles and one wild-type allele. This strain, however, accumulates normal levels of starch granules and lacks phytoglycogen. The total level of isoamylase activity, therefore, is not the major determinant of whether granule production is reduced and phytoglycogen accumulates. Instead, a qualitative property of the isoamylase that is affected by the sta8 mutation is likely to be the critical factor in phytoglycogen production. PMID:11299352

  4. Valorization of Rhizoclonium sp. algae via pyrolysis and catalytic pyrolysis.

    PubMed

    Casoni, Andrés I; Zunino, Josefina; Piccolo, María C; Volpe, María A

    2016-09-01

    The valorization of Rhizoclonium sp. algae through pyrolysis for obtaining bio-oils is studied in this work. The reaction is carried out at 400°C, at high contact time. The bio-oil has a practical yield of 35% and is rich in phytol. Besides, it is simpler than the corresponding to lignocellulosic biomass due to the absence of phenolic compounds. This property leads to a bio-oil relatively stable to storage. In addition, heterogeneous catalysts (Al-Fe/MCM-41, SBA-15 and Cu/SBA-15), in contact with algae during pyrolysis, are analyzed. The general trend is that the catalysts decrease the concentration of fatty alcohols and other high molecular weight products, since their mild acidity sites promote degradation reactions. Thus, the amount of light products increases upon the use of the catalysts. Particularly, acetol concentration in the bio-oils obtained from the catalytic pyrolysis with SBA-15 and Cu/SBA-15 is notably high.

  5. Quantitative analysis of the chemotaxis of a green alga, Chlamydomonas reinhardtii, to bicarbonate using diffusion-based microfluidic device

    PubMed Central

    Kwak, Ho Seok; Sung, Young Joon

    2016-01-01

    There is a growing interest in the photosynthetic carbon fixation by microalgae for the production of valuable products from carbon dioxide (CO2). Microalgae are capable of transporting bicarbonate (HCO3−), the most abundant form of inorganic carbon species in the water, as a source of CO2 for photosynthesis. Despite the importance of HCO3− as the carbon source, little is known about the chemotactic response of microalgae to HCO3−. Here, we showed the chemotaxis of a model alga, Chlamydomonas reinhardtii, towards HCO3− using an agarose gel-based microfluidic device with a flow-free and stable chemical gradient during the entire assay period. The device was validated by analyzing the chemotactic responses of C. reinhardtii to the previously known chemoattractants (NH4Cl and CoCl2) and chemotactically neutral molecule (NaCl). We found that C. reinhardtii exhibited the strongest chemotactic response to bicarbonate at the concentration of 26 mM in a microfluidic device. The chemotactic response to bicarbonate showed a circadian rhythm with a peak during the dark period and a valley during the light period. We also observed the changes in the chemotaxis to bicarbonate by an inhibitor of bicarbonate transporters and a mutation in CIA5, a transcriptional regulator of carbon concentrating mechanism, indicating the relationship between chemotaxis to bicarbonate and inorganic carbon metabolism in C. reinhardtii. To the best of our knowledge, this is the first report of the chemotaxis of C. reinhardtii towards HCO3−, which contributes to the understanding of the physiological role of the chemotaxis to bicarbonate and its relevance to inorganic carbon utilization. PMID:26958101

  6. In silico analysis of the sequence features responsible for alternatively spliced introns in the model green alga Chlamydomonas reinhardtii.

    PubMed

    Raj-Kumar, Praveen-Kumar; Vallon, Olivier; Liang, Chun

    2017-03-31

    Alternatively spliced introns are the ones that are usually spliced but can be occasionally retained in a transcript isoform. They are the most frequently used alternative splice form in plants (~50% of alternative splicing events). Chlamydomonas reinhardtii, a unicellular alga, is a good model to understand alternative splicing (AS) in plants from an evolutionary perspective as it diverged from land plants a billion years ago. Using over 7 million cDNA sequences from both pyrosequencing and Sanger sequencing, we found that a much higher percentage of genes (~20% of multi-exon genes) undergo AS than previously reported (3-5%). We found a full component of SR and SR-like proteins possibly involved in AS. The most prevalent type of AS event (40%) was retention of introns, most of which were supported by multiple cDNA evidence (72%) while only 20% of them have coding capacity. By comparing retained and constitutive introns, we identified sequence features potentially responsible for the retention of introns, in the framework of an "intron definition" model for splicing. We find that retained introns tend to have a weaker 5' splice site, more Gs in their poly-pyrimidine tract and a lesser conservation of nucleotide 'C' at position -3 of the 3' splice site. In addition, the sequence motifs found in the potential branch-point region differed between retained and constitutive introns. Furthermore, the enrichment of G-triplets and C-triplets among the first and last 50 nt of the introns significantly differ between constitutive and retained introns. These could serve as intronic splicing enhancers. All the alternative splice forms can be accessed at http://bioinfolab.miamioh.edu/cgi-bin/PASA_r20140417/cgi-bin/status_report.cgi?db=Chre_AS .

  7. Rhodovulum algae sp. nov., isolated from an algal mat.

    PubMed

    Ramaprasad, E V V; Tushar, L; Dave, Bharti; Sasikala, Ch; Ramana, Ch V

    2016-09-01

    A reddish-brown-pigmented, phototrophic bacterium, designated strain JA877T, was isolated from a brown algae mat sample collected from Jalandhar beach, Gujarat, India. On the basis of the 16S rRNA gene sequence, strain JA877T belongs to the class Alphaproteobacteria and is closely related to the type strains Rhodovulum viride JA756T (99.0 %), Rhodovulum sulfidophilum Hansen W4T (98.9 %), Rhodovulumvisakhapatnamense JA181T (98.8 %),Rhodovulum kholense JA297T (97.5 %) and Rhodovulum salis JA746T (97.0). However, strain JA877T showed only 20-45 % relatedness with its phylogenetic neighbours and had a ∆Tm between 5.8 and 7.0 °C. The major respiratory quinone was ubiquinone-10 (Q10), and the polar lipid profile was composed of the major components phosphatidylglycerol, phosphatidylethanolamine, an unidentified phospholipid, two unidentified sulfolipids and five unidentified lipids. The major fatty acids were C18 : 1ω5c, C18 : 1ω7c/C18 : 1ω6c, C16 : 0 and C18 : 0. The DNA G+C content was 64.5 mol%. On the basis of 16S rRNA gene sequence analysis, physiological data, and chemotaxonomic and molecular differences, strain JA877T is significantly different from other species of the genus Rhodovulum and represents a novel species, for which the name Rhodovulum algae sp. nov. is proposed. The type strain is JA877T (=LMG 29228T= KCTC 42963T).

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

    PubMed

    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.

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

    PubMed Central

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

    2016-01-01

    Snow algae and green algae living in aeroterrestrial habitats are ideal obbjects 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 charopyhte green algae (Zygnema sp., Zygogonium ericetorum and Klebsormidium crenulatum). The hyperspectral microscopic mapping and imaging technique allowed us to acquire total absorbance spectra of these microalgae in the waveband of 400-900 nm. Particularly in Chlamydomonas nivalis and Chlainomonas sp., a high absorbance in the wave band of 400-550 nm was observed, due to naturally occurring secondary carotenoids; in Chloromonas sp. and in the charopyhte algae this 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 not change the spectral properties, in the charopyhte algae the condensation of the cytoplasm and plastids increased the absorbance in the lower waveband of 400 – 500 nm. 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. PMID:27442511

  10. Sulfitobacter undariae sp. nov., isolated from a brown algae reservoir.

    PubMed

    Park, Sooyeon; Jung, Yong-Taek; Won, Sung-Min; Park, Ji-Min; Yoon, Jung-Hoon

    2015-05-01

    A Gram-stain-negative, aerobic, non-spore-forming, non-flagellated and coccoid, ovoid or rod-shaped bacterial strain, W-BA2(T), was isolated from a brown algae reservoir in Wando of South Korea. Strain W-BA2(T) grew optimally at 25 °C, at pH 7.0-8.0 and in the presence of approximately 2.0-3.0% (w/v) NaCl. Phylogenetic trees based on 16S rRNA gene sequences revealed that strain W-BA2(T) fell within the clade comprising the type strains of species of the genus Sulfitobacter , clustering coherently with the type strains of Sulfitobacter donghicola and Sulfitobacter guttiformis showing sequence similarity values of 98.0-98.1%. Sequence similarities to the type strains of the other species of the genus Sulfitobacter were 96.0-97.4%. Strain W-BA2(T) contained Q-10 as the predominant ubiquinone and C18 : 1ω7c as the major fatty acid. The major polar lipids of strain W-BA2(T) were phosphatidylcholine, phosphatidylglycerol, phosphatidylethanolamine, one unidentified aminolipid and one unidentified lipid. The DNA G+C content of strain W-BA2(T) was 55.0 mol% and its DNA-DNA relatedness values with the type strains of Sulfitobacter donghicola , Sulfitobacter guttiformis and Sulfitobacter mediterraneus were 16-23%. The differential phenotypic properties, together with the phylogenetic and genetic distinctiveness, revealed that strain W-BA2(T) is separated from other species of the genus Sulfitobacter . On the basis of the data presented, strain W-BA2(T) is considered to represent a novel species of the genus Sulfitobacter, for which the name Sulfitobacter undariae sp. nov. is proposed. The type strain is W-BA2(T) ( = KCTC 42200(T) = NBRC 110523(T)).

  11. Marinagarivorans algicola gen. nov., sp. nov., isolated from marine algae.

    PubMed

    Guo, Ling-Yun; Li, Dong-Qi; Sang, Jin; Chen, Guan-Jun; Du, Zong-Jun

    2016-01-27

    Novel agar-degrading, Gram-staining-negative, motile, heterotrophic, facultatively anaerobic and pale yellow-pigmented bacterial strains, designated Z1T and JL1, were isolated from marine algae Gelidium amansii (Lamouroux) and Gracilaria verrucosa, respectively. Growth of the isolates was optimal at 28-30 °C, pH 7.0-7.5 and 1-3% (w/v) NaCl. Both strains contained Q-8 as the sole respiratory quinone. The major cellular fatty acids in strain Z1T were C18:1 ω7c, C16:0 and summed feature 3 (C16:1 ω7c and/or iso-C15:0 2-OH). The predominant polar lipids in strain Z1T were phosphatidylethanolamine (PE), phosphatidylglycerol (PG) and aminolipid (AL). The genomic DNA G+C content of both strains was 45.1 mol%. Strains Z1T and JL1 were closely related, with 99.9% 16S rRNA gene sequence similarity. The average nucleotide identity (ANI) value between strains Z1T and JL1 was 99.3%. Phylogenetic analysis based on 16S rRNA gene sequences revealed that strains Z1T and JL1 form a distinct phyletic line within the class Gammaproteobacteria, with less than 92.3% similarity to their closest relatives. Based on data from the current polyphasic study, the isolates are proposed to belong to a new genus and species designated Marinagarivorans algicola gen. nov., sp. nov. The type strain of Marinagarivorans algicola is Z1T (=ATCC BAA-2617T=CICC 10859T).

  12. Chloroplast-mediated regulation of CO2-concentrating mechanism by Ca2+-binding protein CAS in the green alga Chlamydomonas reinhardtii

    PubMed Central

    Wang, Lianyong; Yamano, Takashi; Takane, Shunsuke; Niikawa, Yuki; Toyokawa, Chihana; Ozawa, Shin-ichiro; Tokutsu, Ryutaro; Takahashi, Yuichiro; Minagawa, Jun; Kanesaki, Yu; Yoshikawa, Hirofumi; Fukuzawa, Hideya

    2016-01-01

    Aquatic photosynthetic organisms, including the green alga Chlamydomonas reinhardtii, induce a CO2-concentrating mechanism (CCM) to maintain photosynthetic activity in CO2-limiting conditions by sensing environmental CO2 and light availability. Previously, a novel high-CO2–requiring mutant, H82, defective in the induction of the CCM, was isolated. A homolog of calcium (Ca2+)-binding protein CAS, originally found in Arabidopsis thaliana, was disrupted in H82 cells. Although Arabidopsis CAS is reported to be associated with stomatal closure or immune responses via a chloroplast-mediated retrograde signal, the relationship between a Ca2+ signal and the CCM associated with the function of CAS in an aquatic environment is still unclear. In this study, the introduction of an intact CAS gene into H82 cells restored photosynthetic affinity for inorganic carbon, and RNA-seq analyses revealed that CAS could function in maintaining the expression levels of nuclear-encoded CO2-limiting–inducible genes, including the HCO3– transporters high-light activated 3 (HLA3) and low-CO2–inducible gene A (LCIA). CAS changed its localization from dispersed across the thylakoid membrane in high-CO2 conditions or in the dark to being associated with tubule-like structures in the pyrenoid in CO2-limiting conditions, along with a significant increase of the fluorescent signals of the Ca2+ indicator in the pyrenoid. Chlamydomonas CAS had Ca2+-binding activity, and the perturbation of intracellular Ca2+ homeostasis by a Ca2+-chelator or calmodulin antagonist impaired the accumulation of HLA3 and LCIA. These results suggest that Chlamydomonas CAS is a Ca2+-mediated regulator of CCM-related genes via a retrograde signal from the pyrenoid in the chloroplast to the nucleus. PMID:27791081

  13. Isolation of a bacterial strain, Acinetobacter sp. from centrate wastewater and study of its cooperation with algae in nutrients removal.

    PubMed

    Liu, Hui; Lu, Qian; Wang, Qin; Liu, Wen; Wei, Qian; Ren, Hongyan; Ming, Caibing; Min, Min; Chen, Paul; Ruan, Roger

    2017-03-22

    Algae were able to grow healthy on bacteria-containing centrate wastewater in a pilot-scale bioreactor. The batch experiment indicated that the co-cultivation of algae and wastewater-borne bacteria improved the removal efficiencies of chemical oxygen demand and total phosphorus in centrate wastewater to 93.01% and 98.78%, respectively. A strain of beneficial aerobic bacteria, Acinetobacter sp., was isolated and its biochemical characteristics were explored. Synergistic cooperation was observed in the growth of algae and Acinetobacter sp. Removal efficiencies of some nutrients were improved significantly by the co-cultivation of algae and Acinetobacter sp. After treatment, residual nutrients in centrate wastewater reached the permissible discharge limit. The cooperation between algae and Acinetobacter sp. was in part attributed to the exchange of carbon dioxide and oxygen between the algae and bacteria. This synergetic relationship between algae and Acinetobacter sp. provided a promising way to treat the wastewater by improving the nutrients removal and biomass production.

  14. Expression and knockdown of the PEPC1 gene affect carbon flux in the biosynthesis of triacylglycerols by the green alga Chlamydomonas reinhardtii.

    PubMed

    Deng, Xiaodong; Cai, Jiajia; Li, Yajun; Fei, Xiaowen

    2014-11-01

    The regulation of lipid biosynthesis is important in photosynthetic eukaryotic cells. This regulation is facilitated by the direct synthesis of fatty acids and triacylglycerol (TAG), and by other controls of the main carbon metabolic pathway. In this study, knockdown of the mRNA expression of the Chlamydomonas phosphoenolpyruvate carboxylase isoform 1 (CrPEPC1) gene by RNA interference increased TAG level by 20 % but decreased PEPC activities in the corresponding transgenic algae by 39-50 %. The decrease in CrPEPC1 expression increased the expression of TAG biosynthesis-related genes, such as acyl-CoA:diacylglycerol acyltransferase and phosphatidate phosphatase. Conversely, CrPEPC1 over-expression decreased TAG level by 37 % and increased PEPC activities by 157-184 %. These observations suggest that the lipid content of algal cells can be controlled by regulating the CrPEPC1 gene.

  15. Algibacter undariae sp. nov., isolated from a brown algae reservoir.

    PubMed

    Park, Sooyeon; Lee, Jung-Sook; Lee, Keun-chul; Yoon, Jung-Hoon

    2013-10-01

    A Gram-stain-negative, non-flagellated, rod-shaped bacterial strain able to move by gliding, designated WS-MY9(T), was isolated from a brown algae reservoir in South Korea. Strain WS-MY9(T) grew optimally at 25 °C, at pH 7.0-8.0 and in the presence of 2 % (w/v) NaCl. A neighbour-joining phylogenetic tree based on 16S rRNA gene sequences showed that strain WS-MY9(T) clustered with the type strain of Algibacter lectus with a bootstrap resampling value of 100 %. Strain WS-MY9(T) exhibited 16S rRNA gene sequence similarity values of 98.5 and 96.7 % to the type strains of A. lectus and Algibacter mikhailovii, respectively, and less than 96.1 % sequence similarity to other members of the family Flavobacteriaceae. Strain WS-MY9(T) contained MK-6 as the predominant menaquinone and anteiso-C15 : 0, iso-C17 : 0 3-OH, iso-C15 : 1 G and iso-C15 : 0 as the major fatty acids. The major polar lipids of strain WS-MY9(T) were phosphatidylethanolamine and two unidentified lipids. The DNA G+C content of strain WS-MY9(T) was 35.0 mol% and its DNA-DNA relatedness value with A. lectus KCTC 12103(T) was 15 %. The phylogenetic and genetic distinctiveness and differential phenotypic properties revealed that strain WS-MY9(T) is separate from the two recognized species of the genus Algibacter. On the basis of the data presented, strain WS-MY9(T) represents a novel species of the genus Algibacter, for which the name Algibacter undariae sp. nov. is proposed. The type strain is WS-MY9(T) ( = KCTC 32259(T) = CCUG 63684(T)).

  16. Lacinutrix undariae sp. nov., isolated from a brown algae reservoir.

    PubMed

    Park, Sooyeon; Park, Ji-Min; Jung, Yong-Taek; Kang, Chul-Hyung; Yoon, Jung-Hoon

    2015-08-01

    A Gram-stain-negative, aerobic, non-flagellated, non-gliding and ovoid or rod-shaped bacterium, designated strain W-BA8T, was isolated from a brown algae reservoir on the South Sea, South Korea, and subjected to a polyphasic taxonomic approach. Strain W-BA8T grew optimally at 25 °C, at pH 7.0-7.5 and in the presence of 1.0-2.0% (w/v) NaCl. Neighbour-joining and maximum-parsimony phylogenetic trees based on 16S rRNA gene sequences showed that strain W-BA8T clustered with the type strains of species of the genus Lacinutrix. Strain W-BA8T exhibited 16S rRNA gene sequence similarity values of 94.9-96.5% to the type strains of Lacinutrix species and of less than 95.8% to the type strains of other recognized species. Strain W-BA8T contained MK-6 as the predominant menaquinone and iso-C15 : 0, iso-C15 : 1 G, iso-C15 : 0 3-OH and iso-C17 : 0 3-OH as major fatty acids. The polar lipid profile of strain W-BA8T contained phosphatidylethanolamine, two unidentified lipids and one unidentified glycolipid as major components. The DNA G+C content of strain W-BA8T was 35 mol%. Differential phenotypic properties, together with phylogenetic distinctiveness, revealed that strain W-BA8T is separated from other species of the genus Lacinutrix. On the basis of the data presented, strain W-BA8T is considered to represent a novel species of the genus Lacinutrix, for which the name Lacinutrix undariae sp. nov. is proposed. The type strain is W-BA8T ( = KCTC 42176T = CECT 8671T).

  17. Differences in nutrient uptake capacity of the benthic filamentous algae Cladophora sp., Klebsormidium sp. and Pseudanabaena sp. under varying N/P conditions.

    PubMed

    Liu, Junzhuo; Vyverman, Wim

    2015-03-01

    The N/P ratio of wastewater can vary greatly and directly affect algal growth and nutrient removal process. Three benthic filamentous algae species Cladophora sp., Klebsormidium sp. and Pseudanabaena sp. were isolated from a periphyton bioreactor and cultured under laboratory conditions on varying N/P ratios to determine their ability to remove nitrate and phosphorus. The N/P ratio significantly influenced the algal growth and phosphorus uptake process. Appropriate N/P ratios for nitrogen and phosphorus removal were 5-15, 7-10 and 7-20 for Cladophora sp., Klebsormidium sp. and Pseudanabaena sp., respectively. Within these respective ranges, Cladophora sp. had the highest biomass production, while Pseudanabaena sp. had the highest nitrogen and phosphorus contents. This study indicated that Cladophora sp. had a high capacity of removing phosphorus from wastewaters of low N/P ratio, and Pseudanabaena sp. was highly suitable for removing nitrogen from wastewaters with high N/P ratio.

  18. Whole-Genome Resequencing Reveals Extensive Natural Variation in the Model Green Alga Chlamydomonas reinhardtii[OPEN

    PubMed Central

    Hazzouri, Khaled M.; Rosas, Ulises; Bahmani, Tayebeh; Nelson, David R.; Abdrabu, Rasha; Harris, Elizabeth H.; Salehi-Ashtiani, Kourosh; Purugganan, Michael D.

    2015-01-01

    We performed whole-genome resequencing of 12 field isolates and eight commonly studied laboratory strains of the model organism Chlamydomonas reinhardtii to characterize genomic diversity and provide a resource for studies of natural variation. Our data support previous observations that Chlamydomonas is among the most diverse eukaryotic species. Nucleotide diversity is ∼3% and is geographically structured in North America with some evidence of admixture among sampling locales. Examination of predicted loss-of-function mutations in field isolates indicates conservation of genes associated with core cellular functions, while genes in large gene families and poorly characterized genes show a greater incidence of major effect mutations. De novo assembly of unmapped reads recovered genes in the field isolates that are absent from the CC-503 assembly. The laboratory reference strains show a genomic pattern of polymorphism consistent with their origin as the recombinant progeny of a diploid zygospore. Large duplications or amplifications are a prominent feature of laboratory strains and appear to have originated under laboratory culture. Extensive natural variation offers a new source of genetic diversity for studies of Chlamydomonas, including naturally occurring alleles that may prove useful in studies of gene function and the dissection of quantitative genetic traits. PMID:26392080

  19. Microbiota Influences Morphology and Reproduction of the Brown Alga Ectocarpus sp.

    PubMed Central

    Tapia, Javier E.; González, Bernardo; Goulitquer, Sophie; Potin, Philippe; Correa, Juan A.

    2016-01-01

    Associated microbiota play crucial roles in health and disease of higher organisms. For macroalgae, some associated bacteria exert beneficial effects on nutrition, morphogenesis and growth. However, current knowledge on macroalgae–microbiota interactions is mostly based on studies on green and red seaweeds. In this study, we report that when cultured under axenic conditions, the filamentous brown algal model Ectocarpus sp. loses its branched morphology and grows with a small ball-like appearance. Nine strains of periphytic bacteria isolated from Ectocarpus sp. unialgal cultures were identified by 16S rRNA sequencing, and assessed for their effect on morphology, reproduction and the metabolites secreted by axenic Ectocarpus sp. Six of these isolates restored morphology and reproduction features of axenic Ectocarpus sp. Bacteria-algae co-culture supernatants, but not the supernatant of the corresponding bacterium growing alone, also recovered morphology and reproduction of the alga. Furthermore, colonization of axenic Ectocarpus sp. with a single bacterial isolate impacted significantly the metabolites released by the alga. These results show that the branched typical morphology and the individuals produced by Ectocarpus sp. are strongly dependent on the presence of bacteria, while the bacterial effect on the algal exometabolome profile reflects the impact of bacteria on the whole physiology of this alga. PMID:26941722

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

    PubMed Central

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

    2016-01-01

    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. PMID:27171077

  1. Algae sense exact temperatures: small heat shock proteins are expressed at the survival threshold temperature in Cyanidioschyzon merolae and Chlamydomonas reinhardtii.

    PubMed

    Kobayashi, Yusuke; Harada, Naomi; Nishimura, Yoshiki; Saito, Takafumi; Nakamura, Mami; Fujiwara, Takayuki; Kuroiwa, Tsuneyoshi; Misumi, Osami

    2014-09-29

    The primitive red alga Cyanidioschyzon merolae inhabits acidic hot springs and shows robust resistance to heat shock treatments up to 63 °C. Microarray analysis was performed to identify the key genes underlying the high temperature tolerance of this organism. Among the upregulated genes that were identified, we focused on two small heat shock proteins (sHSPs) that belong to a unique class of HSP families. These two genes are located side by side in an inverted repeat orientation on the same chromosome and share a promoter. These two genes were simultaneously and rapidly upregulated in response to heat shock treatment (>1,000-fold more than the control). Interestingly, upregulation appeared to be triggered not by a difference in temperatures, but rather by the absolute temperature. Similar sHSP structural genes have been reported in the green alga Chlamydomonas reinhardtii, but the threshold temperature for the expression of these sHSP-encoding genes in Ch. reinhardtii was different from the threshold temperature for the expression of the sHSP genes from Cy. merolae. These results indicate the possible importance of an absolute temperature sensing system in the evolution and tolerance of high-temperature conditions among unicellular microalgae.

  2. Seasonal and diel changes in photosynthetic activity of the snow alga Chlamydomonas nivalis (Chlorophyceae) from Svalbard determined by pulse amplitude modulation fluorometry.

    PubMed

    Stibal, Marek; Elster, Josef; Sabacká, Marie; Kastovská, Klára

    2007-02-01

    The seasonal and diel dynamics of the physiological state and photosynthetic activity of the snow alga Chlamydomonas nivalis were investigated in a snowfield in Svalbard. The snow surface represents an environment with very high irradiation intensities along with stable low temperatures close to freezing point. Photosynthetic activity was measured using pulse amplitude modulation fluorometry. Three types of cell (green biflagellate vegetative cells, orange spores clustered by means of mucilaginous sheaths, and purple spores with thick cell walls) were found, all of them photosynthetically active. The pH of snow ranged between 5.0 and 7.5, and the conductivity ranged between 5 and 75 microS cm(-1). The temperature of snow was stable (-0.1 to +0.1 degrees C), and the incident radiation values ranged from 11 to 1500 micromol photons m(-2) s(-1). The photosynthetic activity had seasonal and diel dynamics. The Fv/Fm values ranged between 0.4 and 0.7, and generally declined over the course of the season. A dynamic response of Fv/Fm to the irradiance was recorded. According to the saturating photon fluence values Ek, the algae may have obtained saturating light as deep as 3 cm in the snow when there were higher-light conditions, whereas they were undersaturated at prevalent low light even if on the surface.

  3. 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.

    PubMed

    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.

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

    PubMed

    Winck, Flavia Vischi; Vischi Winck, Flavia; Arvidsson, Samuel; Riaño-Pachón, Diego Mauricio; Hempel, Sabrina; Koseska, Aneta; Nikoloski, Zoran; Urbina Gomez, David Alejandro; Rupprecht, Jens; Mueller-Roeber, Bernd

    2013-01-01

    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. Our work can

  5. RNA-binding protein DUS16 plays an essential role in primary miRNA processing in the unicellular alga Chlamydomonas reinhardtii.

    PubMed

    Yamasaki, Tomohito; Onishi, Masayuki; Kim, Eun-Jeong; Cerutti, Heriberto; Ohama, Takeshi

    2016-09-20

    Canonical microRNAs (miRNAs) are embedded in duplexed stem-loops in long precursor transcripts and are excised by sequential cleavage by DICER nuclease(s). In this miRNA biogenesis pathway, dsRNA-binding proteins play important roles in animals and plants by assisting DICER. However, these RNA-binding proteins are poorly characterized in unicellular organisms. Here we report that a unique RNA-binding protein, Dull slicer-16 (DUS16), plays an essential role in processing of primary-miRNA (pri-miRNA) transcripts in the unicellular green alga Chlamydomonas reinhardtii In animals and plants, dsRNA-binding proteins involved in miRNA biogenesis harbor two or three dsRNA-binding domains (dsRBDs), whereas DUS16 contains one dsRBD and also an ssRNA-binding domain (RRM). The null mutant of DUS16 showed a drastic reduction in most miRNA species. Production of these miRNAs was complemented by expression of full-length DUS16, but the expression of RRM- or dsRBD-truncated DUS16 did not restore miRNA production. Furthermore, DUS16 is predominantly localized to the nucleus and associated with nascent (unspliced form) pri-miRNAs and the DICER-LIKE 3 protein. These results suggest that DUS16 recognizes pri-miRNA transcripts cotranscriptionally and promotes their processing into mature miRNAs as a component of a microprocessor complex. We propose that DUS16 is an essential factor for miRNA production in Chlamydomonas and, because DUS16 is functionally similar to the dsRNA-binding proteins involved in miRNA biogenesis in animals and land plants, our report provides insight into this mechanism in unicellular eukaryotes.

  6. RNA-binding protein DUS16 plays an essential role in primary miRNA processing in the unicellular alga Chlamydomonas reinhardtii

    PubMed Central

    Onishi, Masayuki; Kim, Eun-Jeong; Cerutti, Heriberto; Ohama, Takeshi

    2016-01-01

    Canonical microRNAs (miRNAs) are embedded in duplexed stem–loops in long precursor transcripts and are excised by sequential cleavage by DICER nuclease(s). In this miRNA biogenesis pathway, dsRNA-binding proteins play important roles in animals and plants by assisting DICER. However, these RNA-binding proteins are poorly characterized in unicellular organisms. Here we report that a unique RNA-binding protein, Dull slicer-16 (DUS16), plays an essential role in processing of primary-miRNA (pri-miRNA) transcripts in the unicellular green alga Chlamydomonas reinhardtii. In animals and plants, dsRNA-binding proteins involved in miRNA biogenesis harbor two or three dsRNA-binding domains (dsRBDs), whereas DUS16 contains one dsRBD and also an ssRNA-binding domain (RRM). The null mutant of DUS16 showed a drastic reduction in most miRNA species. Production of these miRNAs was complemented by expression of full-length DUS16, but the expression of RRM- or dsRBD-truncated DUS16 did not restore miRNA production. Furthermore, DUS16 is predominantly localized to the nucleus and associated with nascent (unspliced form) pri-miRNAs and the DICER-LIKE 3 protein. These results suggest that DUS16 recognizes pri-miRNA transcripts cotranscriptionally and promotes their processing into mature miRNAs as a component of a microprocessor complex. We propose that DUS16 is an essential factor for miRNA production in Chlamydomonas and, because DUS16 is functionally similar to the dsRNA-binding proteins involved in miRNA biogenesis in animals and land plants, our report provides insight into this mechanism in unicellular eukaryotes. PMID:27582463

  7. Dehydroascorbate: a possible surveillance molecule of oxidative stress and programmed cell death in the green alga Chlamydomonas reinhardtii.

    PubMed

    Murik, Omer; Elboher, Ahinoam; Kaplan, Aaron

    2014-04-01

    Chlamydomonas reinhardtii tolerates relatively high H2 O2 levels that induce an array of antioxidant activities. However, rather than rendering the cells more resistant to oxidative stress, the cells become far more sensitive to an additional H2 O2 dose. If H2 O2 is provided 1.5-9 h after an initial dose, it induces programmed cell death (PCD) in the wild-type, but not in the dum1 mutant impaired in the mitochondrial respiratory complex III. This mutant does not exhibit a secondary oxidative burst 4-5 h after the inducing H2 O2 , nor does it activate metacaspase-1 after the second H2 O2 treatment. The intracellular dehydroascorbate level, a product of ascorbate peroxidase, increases under conditions leading to PCD. The addition of dehydroascorbate induces PCD in the wild-type and dum1 cultures, but higher levels are required in dum1 cells, where it is metabolized faster. The application of dehydroascorbate induces the expression of metacaspase-2, which is much stronger than the expression of metacaspase-1. The presence or absence of oxidative stress, in addition to the rise in internal dehydroascorbate, may determine which metacaspase is activated during Chlamydomonas PCD. Cell death is strongly affected by the timing of H2 O2 or dehydroascorbate admission to synchronously grown cultures, suggesting that the cell cycle phase may distinguish cells that perish from those that do not.

  8. Toxicity of silver to two freshwater algae, Chlamydomonas reinhardtii and Pseudokirchneriella sub-capitata, grown under continuous culture conditions: influence of thiosulphate.

    PubMed

    Hiriart-Baer, Véronique P; Fortin, Claude; Lee, Dae-Young; Campbell, Peter G C

    2006-06-15

    In a test of the biotic ligand model (BLM), the uptake and toxicity of silver, in the absence or presence of the inorganic ligand, thiosulphate, were assessed for two freshwater green algae, Chlamydomonas reinhardtii and Pseudokirchneriella sub-capitata, using turbidostat continuous cultures. In the initial experiments, run in the absence of thiosulphate, the influent Ag concentration was varied from 0 to 75 nM in steps; for each influent concentration, silver uptake was calculated and the algal growth rate was determined. Silver uptake rates at low Ag concentrations were similar for both algae (e.g., 14-19 nmolm(-2)h(-1), for influent Ag(+) concentrations of approximately 9 nM) but at higher exposures uptake by P. sub-capitata exceeded that of C. reinhardtii. Despite this higher uptake rate, in the absence of thiosulphate P. sub-capitata was not more sensitive to free silver; 50% growth inhibition was reached at influent free Ag(+) concentrations of 15+/-7 and 22+/-13 nM for C. reinhardtii and P. sub-capitata, respectively. In the second series of experiments, the free Ag(+) concentration was held constant ( approximately 9 nM in the influent; 2-3 nM in the effluent) while the concentration of the silver thiosulphate complex, AgS(2)O(3)(-), was increased from 9 to 90 nM in steps. Under such conditions, the BLM would predict that silver uptake and toxicity should remain constant. On the contrary, both silver uptake and silver toxicity increased, indicating that the anionic silver thiosulphate complex enters the algal cells via a membrane-bound sulphate transporter and contributes to uptake and toxicity. However, for both algae there were indications that silver assimilated in this manner was somewhat less toxic to the algal cell than silver that entered via cation transport only. Physiological indicators of stress revealed possible different intracellular targets for these two freshwater algae, proteins and enzymes for C. reinhardtii and the photosynthetic

  9. Isolation, antimicrobial activity, and metabolites of fungus Cladosporium sp. associated with red alga Porphyra yezoensis.

    PubMed

    Ding, Ling; Qin, Song; Li, Fuchao; Chi, Xiaoyuan; Laatsch, Hartmut

    2008-03-01

    Cladosporium sp. isolate N5 was isolated as a dominant fungus from the healthy conchocelis of Porphyra yezoensis. In the re-infection test, it did not cause any pathogenic symptoms in the alga. Twenty-one cultural conditions were chosen to test its antimicrobial activity in order to obtain the best condition for large-scale fermentation. Phenylacetic acid, p-hydroxyphenylethyl alcohol, and L-beta-phenyllactic acid were isolated from the crude extract as strong antimicrobial compounds and they are the first reported secondary metabolites for the genus Cladosporium. In addition, the Cladosporium sp. produced the reported Porphyra yezoensis growth regulators phenylacetic acid and p-hydroxyphenylacetic acid. No cytotoxicity was found in the brine shrimp lethality test, which indicated that the environmental-friendly Cladosporium sp. could be used as a potential biocontrol agent to protect the alga from pathogens.

  10. Kordia ulvae sp. nov., a bacterium isolated from the surface of green marine algae Ulva sp.

    PubMed

    Qi, Feng; Huang, Zhaobin; Lai, Qiliang; Li, Dengfeng; Shao, Zongze

    2016-04-20

    A novel bacterial strain SC2T was isolated from Ulva sp. a green marine algae. Strain SC2T was Gram-negative, aerobic, rod-shaped and had no flagellum. Oxidase and catalase were positive. Strain SC2T can degrade skim milk, agar, soluble starch, Tween 20 and Tween 80. The optimal salinity and temperature of strain SC2T were 2% and 30 °C, respectively. Phylogenetic analysis based on the 16S rRNA gene indicated that strain SC2T was affiliated to the genus Kordia, with highest sequence similarity to Kordia algicida OT-1T (97.23%), Kordia antarctica IMCC3317T (97.23%) and Kordia jejudonensis SSK3-3T (97.02%); other species of the genus Kordia shared 93.98%-95.78% sequence similarity. The ANI value and the DNA-DNA hybridization estimated value between strain SC2T and three type strains (K. algicida OT-1T, K. antarctica IMCC3317T and K. jejudonensis SSK3-3T) were found to be 79.4%-82.4% and 24.2%-27.0%, respectively. The predominant fatty acids (>5.0%) were C16:0, iso-C15:0, iso-C15:0 3-OH, iso-C17:0 3-OH, summed feature 3 (comprised C16:1 ω7c/C16:1 ω6c), summed feature 8 (comprised C18:1 ω7c/C18:1 ω6c) and summed feature 9 (comprised iso-C17:1 ω9c/C16:0 10-methyl). The respiratory quinone was Menaquinone-6 (MK-6). The polar lipid profile consisted of four unknown lipids, three unidentified phospholipids, one unidentified aminolipid and one phosphatidylethanolamine. The G+C content of the genomic DNA was 34.5 mol%. The combined genotypic and phenotypic data showed that strain SC2T represents a novel species within the genus Kordia, for which the name Kordia ulvae sp. nov. is proposed, with the type strain SC2T (= KCTC 42872T = MCCC 1A01772T = LMG 29123T).

  11. Negative Impact on Growth and Photosynthesis in the Green Alga Chlamydomonas reinhardtii in the Presence of the Estrogen 17α-Ethynylestradiol

    PubMed Central

    Pocock, Tessa; Falk, Stefan

    2014-01-01

    It is well known that estrogenic compounds affect development of fertilized eggs of many species of birds, fish and amphibians through disrupted activity of carbonic anhydrase (CA). The most potent activity comes from the most commonly occurring synthetic sterol, 17α-Ethynylestradiol (EE2). Less is known about the responses of aquatic phytoplankton to these compounds. Here we show for the first time that, in comparision to the control, the addition of 7 µM EE2 reduced the growth rate of the green alga Chlamydomonas reinhardtii by 68% for cells grown at high CO2. When cells were grown in ambient air (low Ci) with a fully activated carbon concentrating mechanism through the induction of CA activity, the growth rates were reduced by as much as 119%. A reduced growth rate could be observed at EE2 concentrations as low as 10 pM. This was accompanied by a reduced maximum capacity for electron transport in photosystem II as determined by a lower FV/FM for low Ci-grown cells, which indicates the involvement of CAH3, a CA specifically located in the thylakoid lumen involved in proton pumping across the thylakoid membranes. These results were in agreement with an observed reduction in the chloroplastic affinity for Ci as shown by a strong increase in the Michaelis-Menten K0.5 for HCO3−. In itself, a lowering of the growth rate of a green alga by addition of the sterol EE2 warrants further investigation into the potential environmental impact by the release of treated waste water. PMID:25310092

  12. Negative impact on growth and photosynthesis in the green alga Chlamydomonas reinhardtii in the presence of the estrogen 17α-ethynylestradiol.

    PubMed

    Pocock, Tessa; Falk, Stefan

    2014-01-01

    It is well known that estrogenic compounds affect development of fertilized eggs of many species of birds, fish and amphibians through disrupted activity of carbonic anhydrase (CA). The most potent activity comes from the most commonly occurring synthetic sterol, 17α-Ethynylestradiol (EE2). Less is known about the responses of aquatic phytoplankton to these compounds. Here we show for the first time that, in comparision to the control, the addition of 7 µM EE2 reduced the growth rate of the green alga Chlamydomonas reinhardtii by 68% for cells grown at high CO2. When cells were grown in ambient air (low Ci) with a fully activated carbon concentrating mechanism through the induction of CA activity, the growth rates were reduced by as much as 119%. A reduced growth rate could be observed at EE2 concentrations as low as 10 pM. This was accompanied by a reduced maximum capacity for electron transport in photosystem II as determined by a lower FV/FM for low Ci-grown cells, which indicates the involvement of CAH3, a CA specifically located in the thylakoid lumen involved in proton pumping across the thylakoid membranes. These results were in agreement with an observed reduction in the chloroplastic affinity for Ci as shown by a strong increase in the Michaelis-Menten K0.5 for HCO3-. In itself, a lowering of the growth rate of a green alga by addition of the sterol EE2 warrants further investigation into the potential environmental impact by the release of treated waste water.

  13. The slow S to M rise of chlorophyll a fluorescence reflects transition from state 2 to state 1 in the green alga Chlamydomonas reinhardtii.

    PubMed

    Kodru, Sireesha; Malavath, Tirupathi; Devadasu, Elsinraju; Nellaepalli, Sreedhar; Stirbet, Alexandrina; Subramanyam, Rajagopal; Govindjee

    2015-08-01

    The green alga Chlamydomonas (C.) reinhardtii is a model organism for photosynthesis research. State transitions regulate redistribution of excitation energy between photosystem I (PS I) and photosystem II (PS II) to provide balanced photosynthesis. Chlorophyll (Chl) a fluorescence induction (the so-called OJIPSMT transient) is a signature of several photosynthetic reactions. Here, we show that the slow (seconds to minutes) S to M fluorescence rise is reduced or absent in the stt7 mutant (which is locked in state 1) in C. reinhardtii. This suggests that the SM rise in wild type C. reinhardtii may be due to state 2 (low fluorescence state; larger antenna in PS I) to state 1 (high fluorescence state; larger antenna in PS II) transition, and thus, it can be used as an efficient and quick method to monitor state transitions in algae, as has already been shown in cyanobacteria (Papageorgiou et al. 1999, 2007; Kaňa et al. 2012). We also discuss our results on the effects of (1) 3-(3,4-dichlorophenyl)-1,4-dimethyl urea, an inhibitor of electron transport; (2) n-propyl gallate, an inhibitor of alternative oxidase (AOX) in mitochondria and of plastid terminal oxidase in chloroplasts; (3) salicylhydroxamic acid, an inhibitor of AOX in mitochondria; and (4) carbonyl cyanide p-trifluoromethoxyphenylhydrazone, an uncoupler of phosphorylation, which dissipates proton gradient across membranes. Based on the data presented in this paper, we conclude that the slow PSMT fluorescence transient in C. reinhardtii is due to the superimposition of, at least, two phenomena: qE dependent non-photochemical quenching of the excited state of Chl, and state transitions.

  14. Rapid Aggregation of Biofuel-Producing Algae by the Bacterium Bacillus sp. Strain RP1137

    PubMed Central

    Powell, Ryan J.

    2013-01-01

    Algal biofuels represent one of the most promising means of sustainably replacing liquid fuels. However, significant challenges remain before alga-based fuels become competitive with fossil fuels. One of the largest challenges is the ability to harvest the algae in an economical and low-energy manner. In this article, we describe the isolation of a bacterial strain, Bacillus sp. strain RP1137, which can rapidly aggregate several algae that are candidates for biofuel production, including a Nannochloropsis sp. This bacterium aggregates algae in a pH-dependent and reversible manner and retains its aggregation ability after paraformaldehyde fixation, opening the possibility for reuse of the cells. The optimal ratio of bacteria to algae is described, as is the robustness of aggregation at different salinities and temperatures. Aggregation is dependent on the presence of calcium or magnesium ions. The efficiency of aggregation of Nannochloropsis oceanica IMET1 is between 70 and 95% and is comparable to that obtained by other means of harvest; however, the rate of harvest is fast, with aggregates forming in 30 s. PMID:23892750

  15. Rapid aggregation of biofuel-producing algae by the bacterium Bacillus sp. strain RP1137.

    PubMed

    Powell, Ryan J; Hill, Russell T

    2013-10-01

    Algal biofuels represent one of the most promising means of sustainably replacing liquid fuels. However, significant challenges remain before alga-based fuels become competitive with fossil fuels. One of the largest challenges is the ability to harvest the algae in an economical and low-energy manner. In this article, we describe the isolation of a bacterial strain, Bacillus sp. strain RP1137, which can rapidly aggregate several algae that are candidates for biofuel production, including a Nannochloropsis sp. This bacterium aggregates algae in a pH-dependent and reversible manner and retains its aggregation ability after paraformaldehyde fixation, opening the possibility for reuse of the cells. The optimal ratio of bacteria to algae is described, as is the robustness of aggregation at different salinities and temperatures. Aggregation is dependent on the presence of calcium or magnesium ions. The efficiency of aggregation of Nannochloropsis oceanica IMET1 is between 70 and 95% and is comparable to that obtained by other means of harvest; however, the rate of harvest is fast, with aggregates forming in 30 s.

  16. Optimizing biodiesel production in marine Chlamydomonas sp. JSC4 through metabolic profiling and an innovative salinity-gradient strategy

    PubMed Central

    2014-01-01

    Background Biodiesel production from marine microalgae has received much attention as microalgae can be cultivated on non-arable land without the use of potable water, and with the additional benefits of mitigating CO2 emissions and yielding biomass. However, there is still a lack of effective operational strategies to promote lipid accumulation in marine microalgae, which are suitable for making biodiesel since they are mainly composed of saturated and monounsaturated fatty acids. Moreover, the regulatory mechanisms involved in lipid biosynthesis in microalgae under environmental stress are not well understood. Results In this work, the combined effects of salinity and nitrogen depletion stresses on lipid accumulation of a newly isolated marine microalga, Chlamydomonas sp. JSC4, were explored. Metabolic intermediates were profiled over time to observe transient changes during the lipid accumulation triggered by the combination of the two stresses. An innovative cultivation strategy (denoted salinity-gradient operation) was also employed to markedly improve the lipid accumulation and lipid quality of the microalga, which attained an optimal lipid productivity of 223.2 mg L-1 d-1 and a lipid content of 59.4% per dry cell weight. This performance is significantly higher than reported in most related studies. Conclusions This work demonstrated the synergistic integration of biological and engineering technologies to develop a simple and effective strategy for the enhancement of oil production in marine microalgae. PMID:25002905

  17. tla1, a DNA insertional transformant of the green alga Chlamydomonas reinhardtii with a truncated light-harvesting chlorophyll antenna size.

    PubMed

    Polle, Juergen E W; Kanakagiri, Sarada-Devi; Melis, Anastasios

    2003-05-01

    DNA insertional mutagenesis and screening of the green alga Chlamydomonas reinhardtii was employed to isolate tla1, a stable transformant having a truncated light-harvesting chlorophyll antenna size. Molecular analysis showed a single plasmid insertion into an open reading frame of the nuclear genome corresponding to a novel gene ( Tla1) that encodes a protein of 213 amino acids. Genetic analysis showed co-segregation of plasmid and tla1 phenotype. Biochemical analyses showed the tla1 mutant to be chlorophyll deficient, with a functional chlorophyll antenna size of photosystem I and photosystem II being about 50% and 65% of that of the wild type, respectively. It contained a correspondingly lower amount of light-harvesting proteins than the wild type and had lower steady-state levels of Lhcb mRNA. The tla1 strain required a higher light intensity for the saturation of photosynthesis and showed greater solar conversion efficiencies and a higher photosynthetic productivity than the wild type under mass culture conditions. Results are discussed in terms of the tla1 mutation, its phenotype, and the role played by the Tla1 gene in the regulation of the photosynthetic chlorophyll antenna size in C. reinhardtii.

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

    SciTech Connect

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

    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.

  19. Sorption of copper(II) ions in the biomass of alga Spirogyra sp.

    PubMed

    Rajfur, Małgorzata; Kłos, Andrzej; Wacławek, Maria

    2012-10-01

    Sorption of copper ions by the alga Spirogyra sp. was investigated to determine the influence of experimental conditions and the methods of sample preparation on the process. The experiments were carried out both under the static and the dynamic conditions. Kinetics and equilibrium parameters of the sorption were evaluated. In addition, the influence was studied of the algae preparation methods on the conductivity of demineralized water in which the algae samples were immersed. The static experiments showed that the sorption of Cu(2+) ions reached equilibrium in about 30 min, with approximately 90% of the ions adsorbed in the initial 15 min. The sorption capacity determined from the Langmuir isotherms appeared highly uncertain (SD=±0.027 mg/g dry mass or ±11%, for the live algae). Under static conditions, the slopes of the Langmuir isotherms depended on the ratio of the alga mass to the volume of solution. The conductometric measurements were proven to be a simple and fast way to evaluate the quality of algae used for the experiments.

  20. Genome-Wide Identification of Regulatory Elements and Reconstruction of Gene Regulatory Networks of the Green Alga Chlamydomonas reinhardtii under Carbon Deprivation

    PubMed Central

    Vischi Winck, Flavia; Arvidsson, Samuel; Riaño-Pachón, Diego Mauricio; Hempel, Sabrina; Koseska, Aneta; Nikoloski, Zoran; Urbina Gomez, David Alejandro; Rupprecht, Jens; Mueller-Roeber, Bernd

    2013-01-01

    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-CO2 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. Our work can

  1. High-efficiency homologous recombination in the oil-producing alga Nannochloropsis sp.

    PubMed

    Kilian, Oliver; Benemann, Christina S E; Niyogi, Krishna K; Vick, Bertrand

    2011-12-27

    Algae have reemerged as potential next-generation feedstocks for biofuels, but strain improvement and progress in algal biology research have been limited by the lack of advanced molecular tools for most eukaryotic microalgae. Here we describe the development of an efficient transformation method for Nannochloropsis sp., a fast-growing, unicellular alga capable of accumulating large amounts of oil. Moreover, we provide additional evidence that Nannochloropsis is haploid, and we demonstrate that insertion of transformation constructs into the nuclear genome can occur by high-efficiency homologous recombination. As examples, we generated knockouts of the genes encoding nitrate reductase and nitrite reductase, resulting in strains that were unable to grow on nitrate and nitrate/nitrite, respectively. The application of homologous recombination in this industrially relevant alga has the potential to rapidly advance algal functional genomics and biotechnology.

  2. Sex determination in Chlamydomonas.

    PubMed

    Goodenough, Ursula; Lin, Huawen; Lee, Jae-Hyeok

    2007-06-01

    The sex-determination system of the unicellular green alga, Chlamydomonas reinhardtii, is governed by genes in the mating-type (MT) locus and entails additional genes located in autosomes. Gene expression is initiated by nitrogen starvation, and cells differentiate into plus or minus gametes within 6h. Reviewed is our current understanding of gametic differentiation and fertilization, initiation of zygote development, and the uniparental inheritance of organelle genomes.

  3. Pyropia plicata sp. nov. (Bangiales, Rhodophyta): naming a common intertidal alga from New Zealand

    PubMed Central

    Nelson, Wendy A.

    2013-01-01

    Abstract A commonly found red alga of the upper intertidal zone of New Zealand rocky coasts is described for the first time as Pyropia plicata sp. nov. This species has been incorrectly known as Porphyra columbina Mont. (now Pyropia columbina (Mont.) W.A.Nelson) for many years. Pyropia plicata is widespread and common, and it is readily distinguished from other species of bladed Bangiales in New Zealand by its distinctive morphology, with pleated blades attached by a central rhizoidal holdfast. PMID:23794933

  4. Pyropia plicata sp. nov. (Bangiales, Rhodophyta): naming a common intertidal alga from New Zealand.

    PubMed

    Nelson, Wendy A

    2013-01-01

    A commonly found red alga of the upper intertidal zone of New Zealand rocky coasts is described for the first time as Pyropia plicata sp. nov. This species has been incorrectly known as Porphyra columbina Mont. (now Pyropia columbina (Mont.) W.A.Nelson) for many years. Pyropia plicata is widespread and common, and it is readily distinguished from other species of bladed Bangiales in New Zealand by its distinctive morphology, with pleated blades attached by a central rhizoidal holdfast.

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

    PubMed

    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.

  6. Algae.

    PubMed

    Raven, John A; Giordano, Mario

    2014-07-07

    Algae frequently get a bad press. Pond slime is a problem in garden pools, algal blooms can produce toxins that incapacitate or kill animals and humans and even the term seaweed is pejorative - a weed being a plant growing in what humans consider to be the wrong place. Positive aspects of algae are generally less newsworthy - they are the basis of marine food webs, supporting fisheries and charismatic marine megafauna from albatrosses to whales, as well as consuming carbon dioxide and producing oxygen. Here we consider what algae are, their diversity in terms of evolutionary origin, size, shape and life cycles, and their role in the natural environment and in human affairs.

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

    PubMed Central

    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

  8. Assembly of the light-harvesting chlorophyll antenna in the green alga Chlamydomonas reinhardtii requires expression of the TLA2-CpFTSY gene.

    PubMed

    Kirst, Henning; García-Cerdán, Jose Gines; Zurbriggen, Andreas; Melis, Anastasios

    2012-02-01

    The truncated light-harvesting antenna2 (tla2) mutant of Chlamydomonas reinhardtii showed a lighter-green phenotype, had a lower chlorophyll (Chl) per-cell content, and higher Chl a/b ratio than corresponding wild-type strains. Physiological analyses revealed a higher intensity for the saturation of photosynthesis and greater P(max) values in the tla2 mutant than in the wild type. Biochemical analyses showed that the tla2 strain was deficient in the Chl a-b light-harvesting complex, and had a Chl antenna size of the photosystems that was only about 65% of that in the wild type. Molecular and genetic analyses showed a single plasmid insertion in the tla2 strain, causing a chromosomal DNA rearrangement and deletion/disruption of five nuclear genes. The TLA2 gene, causing the tla2 phenotype, was cloned by mapping the insertion site and upon complementation with each of the genes that were deleted. Successful complementation was achieved with the C. reinhardtii TLA2-CpFTSY gene, whose occurrence and function in green microalgae has not hitherto been investigated. Functional analysis showed that the nuclear-encoded and chloroplast-localized CrCpFTSY protein specifically operates in the assembly of the peripheral components of the Chl a-b light-harvesting antenna. In higher plants, a cpftsy null mutation inhibits assembly of both the light-harvesting complex and photosystem complexes, thus resulting in a seedling-lethal phenotype. The work shows that cpftsy deletion in green algae, but not in higher plants, can be employed to generate tla mutants. The latter exhibit improved solar energy conversion efficiency and photosynthetic productivity under mass culture and bright sunlight conditions.

  9. RNAi Knock-Down of LHCBM1, 2 and 3 Increases Photosynthetic H2 Production Efficiency of the Green Alga Chlamydomonas reinhardtii

    PubMed Central

    Oey, Melanie; Ross, Ian L.; Stephens, Evan; Steinbeck, Janina; Wolf, Juliane; Radzun, Khairul Adzfa; Kügler, Johannes; Ringsmuth, Andrew K.; Kruse, Olaf; Hankamer, Ben

    2013-01-01

    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. PMID:23613840

  10. Photochemical Performance of the Acidophilic Red Alga Cyanidium sp. in a pH Gradient

    NASA Astrophysics Data System (ADS)

    Kvíderová, Jana

    2012-06-01

    The acidophilic red alga Cyanidium sp. is one of the dominant mat-forming species in the highly acidic waters of Río Tinto, Spain. The culture of Cyanidium sp., isolated from a microbial mat sample collected at Río Tinto, was exposed to 9 different pH conditions in a gradient from 0.5 to 5 for 24 h and its physiological status evaluated by variable chlorophyll a fluorescence kinetics measurements. Maximum quantum yield was determined after 30 min, 1 h, 2 h, 4 h, 6 h and 24 h of exposure after 15 min dark adaptation. The effect of pH on photochemical activity of Cyanidium sp. was observable as early as 30 min after exposure and the pattern remained stable or with only minor modifications for 24 h. The optimum pH ranged from 1.5 to 2.5. A steep decrease of the photochemical activity was observed at pH below 1 even after 30 min of exposure. Although the alga had tolerated the exposure to pH = 1 for at least 6 h, longer (24 h) exposure resulted in reduction of the photochemical activity. At pH above 2.5, the decline was more moderate and its negative effect on photochemistry was less severe. According to the fluorescence measurements, the red alga Cyanidium sp. is well-adapted to prevailing pH at its original locality at Río Tinto, i.e. pH of 1 to 3. The short-term survival in pH < 1.5 may be adaptation to rare exposures to such low pH in the field. The tolerance of pH above 3 could be caused by adaptation to the microenvironment of the inner parts of microbial mats in which Cyanidium sp. usually dominates and where higher pH could occur due to photosynthetic oxygen production.

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

    PubMed

    Röhder, Lena A; Brandt, Tanja; Sigg, Laura; Behra, Renata

    2014-07-01

    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 ∼0mV at physiological pH and agglomerated in exposure media. Phosphate stabilized CeO2 NP at pH 7.5 over 24h. This effect was exploited to test CeO2 NP dispersed in phosphate with a mean size of 140nm and agglomerated in absence of phosphate with a mean size of 2000nm. The level of dissolved cerium(III) in CeO2 NP suspensions was very low and between 0.1 and 27nM 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±0.84μM for wild type and EC50 of 6.3±0.53μM 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 CeO2 NP caused a slight decrease of photosynthetic yield at the highest concentrations (100μM), while no effect was observed for dispersed CeO2 NP. The low toxicity of agglomerated CeO2 NP was attributed quantitatively to Ce(3+) ions co-occurring in the nanoparticle suspension whereas for dispersed CeO2 NP, dissolved Ce(3+) was precipitated with phosphate and not bioavailable. Furthermore CeO2 NP did not affect the intracellular ROS level. The cell wall free mutant and wild type of C

  12. Metabolic and gene expression changes triggered by nitrogen deprivation in the photoautotrophically grown microalgae Chlamydomonas reinhardtii and Coccomyxa sp. C-169.

    PubMed

    Msanne, Joseph; Xu, Di; Konda, Anji Reddy; Casas-Mollano, J Armando; Awada, Tala; Cahoon, Edgar B; Cerutti, Heriberto

    2012-03-01

    Microalgae are emerging as suitable feedstocks for renewable biofuel production. Characterizing the metabolic pathways involved in the biosynthesis of energy-rich compounds, such as lipids and carbohydrates, and the environmental factors influencing their accumulation is necessary to realize the full potential of these organisms as energy resources. The model green alga Chlamydomonas reinhardtii accumulates significant amounts of triacylglycerols (TAGs) under nitrogen starvation or salt stress in medium containing acetate. However, since cultivation of microalgae for biofuel production may need to rely on sunlight as the main source of energy for biomass synthesis, metabolic and gene expression changes occurring in Chlamydomonas and Coccomyxa subjected to nitrogen deprivation were examined under strictly photoautotrophic conditions. Interestingly, nutrient depletion triggered a similar pattern of early synthesis of starch followed by substantial TAG accumulation in both of these fairly divergent green microalgae. A marked decrease in chlorophyll and protein contents was also observed, including reduction in ribosomal polypeptides and some key enzymes for CO₂ assimilation like ribulose-1,5-bisphosphate carboxylase/oxygenase. These results suggest that turnover of nitrogen-rich compounds such as proteins may provide carbon/energy for TAG biosynthesis in the nutrient deprived cells. In Chlamydomonas, several genes coding for diacylglycerol:acyl-CoA acyltransferases, catalyzing the acylation of diacylglycerol to TAG, displayed increased transcript abundance under nitrogen depletion but, counterintuitively, genes encoding enzymes for de novo fatty acid synthesis, such as 3-ketoacyl-ACP synthase I, were down-regulated. Understanding the interdependence of these anabolic and catabolic processes and their regulation may allow the engineering of algal strains with improved capacity to convert their biomass into useful biofuel precursors.

  13. Some Nutritional Characteristics of a Naturally Occurring Alga (Microcystis sp.) in a Guatemalan Lake

    PubMed Central

    de la Fuente, Gabriel; Flores, Antonio; Molina, Mario R.; Almengor, Leticia; Bressani, Ricardo

    1977-01-01

    The nutritional characteristics of an alga (Microcystis sp.) that occurs naturally in a Guatemalan lake were determined. The sun-dried material proved to have a high protein content (55.6%) and to be a possible good source of calcium and phosphorus (1, 169.1 and 633.4 mg/100 mg, respectively). Amino acid analysis showed that total sulfur amino acids were the most deficient ones, giving a protein score of 42 to the material. The in vitro protein digestibility of the material was 69.5%. Biological trials demonstrated that when the material was offered as the only protein source, very low consumption and a high mortality rate were obtained whether or not the diet was supplemented with 0.4% dl-methionine. However, when the material supplied 25% of the total protein of a corn-algae diet, the protein quality of the cereal was significantly improved (P < 0.05). PMID:16345191

  14. Chlamydomonas: A Model Green Plant.

    ERIC Educational Resources Information Center

    Sheffield, E.

    1985-01-01

    Discusses the instructional potential of Chlamydomonas in providing a basis for a range of experimental investigations to illustrate basic biological phenomena. Describes the use of this algae genus in studies of population growth, photosynthesis, and mating behavior. Procedures for laboratory exercises are included. (ML)

  15. Hylodesmus singaporensis gen. et sp. nov., a new autosporic subaerial green alga (Scenedesmaceae, Chlorophyta) from Singapore.

    PubMed

    Eliás, Marek; Nemcová, Yvonne; Skaloud, Pavel; Neustupa, Jirí; Kaufnerová, Veronika; Sejnohová, Lenka

    2010-05-01

    The algal flora of subaerial habitats in the tropics remains largely unexplored, despite the fact that it potentially encompasses a wealth of new evolutionary diversity. Here we present a detailed morphological and molecular characterization of an autosporic coccoid green alga isolated from decaying wood in a natural forest in Singapore. Depending on culture conditions, this alga formed globular to irregularly oval solitary cells. Autosporulation was the only mode of reproduction observed. The cell periphery was filled with numerous vacuoles, and a single parietal chloroplast contained a conspicuous pyrenoid surrounded by a bipartite starch envelope. The cell wall was composed of a thick inner layer and a thin trilaminar outer layer, and the cell surface was ornamented with a few delicate ribs. Phylogenetic analyses of 18S rRNA gene sequences placed our strain in the family Scenedesmaceae (Sphaeropleales, Chlorophyceae) as a strongly supported sister branch of the genus Desmodesmus. Analyses of an alternative phylogenetic marker widely used for the Scenedesmaceae, the ITS2 region, confirmed that the strain is distinct from any scenedesmacean alga sequenced to date, but is related to the genus Desmodesmus, despite lacking the defining phenotypic features of Desmodesmus (cell wall with four sporopolleninic layers ornamented with peculiar submicroscopic structures). Collectively, our results establish that we identified a novel, previously undocumented, evolutionary lineage of scenedesmacean algae necessitating its description as a new species in a new genus. We propose it be named Hylodesmus singaporensis gen. et sp. nov. A cryopreserved holotype specimen has been deposited into the Culture Collection of Algae of Charles University in Prague, Czech Republic (CAUP) as CAUP C-H8001.

  16. New α-glucosidase inhibitors from marine algae-derived Streptomyces sp. OUCMDZ-3434

    PubMed Central

    Chen, Zhengbo; Hao, Jiejie; Wang, Liping; Wang, Yi; Kong, Fandong; Zhu, Weiming

    2016-01-01

    Wailupemycins H (1) and I (2) with a new skeleton coupled two 6-(2-phenylnaphthalene-1-yl)pyrane-2-one nuclei to a –CH2– linkage were identified from the culture of Streptomyces sp. OUCMDZ-3434 associated with the marine algae, Enteromorpha prolifera. Compounds 1 and 2 are two new α-glucosidase inhibitors with the Ki/IC50 values of 16.8/19.7 and 6.0/8.3 μM, respectively. In addition, the absolute configurations of wailupemycins D (3) and E (4) are also resolved in this paper for the first time. PMID:26822662

  17. Cellular Fe-hydroxides and heavy metal sorption in Euglena sp. (algae): implications for biomineralization

    SciTech Connect

    Mann, H.; Beveridge, T.O. Fyfe, W.S.; Tazaki, K.

    1985-01-01

    STEM imagery and electron diffraction patterns of Euglena sp. reveal pronounced intra and cellular-membrane aggregates of Fe-hydroxides (some lepidocrocite), in natural communities from tailings waters, Elliott Lake, Ontario. Pure isolates of Euglena sp. contain 40-70% Fe by dry weight and in addition average Al 28,000 ppm, Sr 150, Ba 40, Zn 150, Mn 250, Ni 120, Pb 1600, Th 70, Cu 200 and U 180. In tailings waters, Fe solute concentrations average 560 ppm and U 50 ppb. Concentration factors for Fe, Ba, Zn, Mn, Ti, V, Ni, Pb, Cr, Ag, Co and Cu in algae referenced to average world river waters are greater than or equal to 10/sup 6/. These results endorse the premise that microorganisms mediate transfer of many solutes between the hydrosphere and sedimentary regime.

  18. 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.

    PubMed

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

    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.

  19. Structural characteristics and biological activity of Fucoidans from the brown algae Alaria sp. and Saccharina japonica of different reproductive status.

    PubMed

    Vishchuk, Olesya S; Tarbeeva, Dariya V; Ermakova, Svetlana P; Zvyagintseva, Tatyana N

    2012-04-01

    Structural characteristics and the antitumor activity of fucoidans isolated from vegetative and reproductive tissue of the brown algae Alaria sp. and Saccharina japonica were studied. The reproductive status of the brown algae affected the yield of fucoidans and their structural characteristics. The fucoidan yield was 5.7% (w/w on the basis of the dried algae weight) for fertile and 3.8% for sterile Alaria sp. and 1.42 and 0.71% for fertile and sterile S. japonica, respectively. The fucoidans from fertile Alaria sp. and S. japonica had a slightly higher degree of sulfation and a somewhat more homogeneous monosaccharide composition, with predominate amounts of fucose and galactose, than those isolated from sterile algae tissue. The fucoidans from both the sterile and fertile brown algae tissue tested possessed selective cytotoxicity towards human breast cancer (T-47D) and melanoma (RPMI-7951) cell lines, but not to normal mouse epidermal cells (JB6 Cl41), and effectively inhibited the proliferation and colony formation of the breast cancer and melanoma cell lines. The fucoidans from reproductive tissue of brown algae possessed higher antitumor activity than those from vegetative plants.

  20. Toxicity and transformation of fenamiphos and its metabolites by two micro algae Pseudokirchneriella subcapitata and Chlorococcum sp.

    PubMed

    Cáceres, Tanya; Megharaj, Mallavarapu; Naidu, Ravi

    2008-07-15

    The acute toxicity of an organophosphorous pesticide, fenamiphos and its metabolites, fenamiphos sulfoxide (FSO), fenamiphos sulfone (FSO(2)), fenamiphos phenol (FP), fenamiphos sulfoxide phenol (FSOP) and fenamiphos sulfone phenol (FSO(2)P), to the aquatic alga Pseudokirchneriella subcapitata and the terrestrial alga Chlorococcum sp. was studied. The toxicity followed the order: fenamiphos phenol>fenamiphos sulfone phenol>fenamiphos sulfoxide phenol>fenamiphos. The oxidation products of fenamiphos, FSO and FSO(2) were not toxic to the algal species up to 100 mg L(-1). Both algae were able to transform fenamiphos, FSO and FSO(2), while the phenols were found to be stable in the incubation media. Bioaccumulation of both fenamiphos and its metabolites was observed in the case of Chlorococcum sp. while only metabolites were accumulated in P. subcapitata. This study demonstrates that (i) the hydrolysis products of fenamiphos, FSOP and FSO(2)P are more toxic to both fresh water and soil algae than their parent chemicals, (ii) further fenamiphos can be transformed and bioconcentrated by these algae. Therefore, contamination of natural environments such as waterbodies with fenamiphos or its metabolites can have adverse impacts on the food chain and associated biota (especially to the primary consumers such as Daphnia) since algae are the primary producers located at the base of the food chain. Further, the finding that the fenamiphos phenols are more toxic to algae highlights the need to consider the transformation products in ecological risk assessment of fenamiphos.

  1. Defluviitalea phaphyphila sp. nov., a Novel Thermophilic Bacterium That Degrades Brown Algae

    PubMed Central

    Ji, Shi-Qi; Wang, Bing; Lu, Ming

    2015-01-01

    Brown algae are one of the largest groups of oceanic primary producers for CO2 removal and carbon sinks for coastal regions. However, the mechanism for brown alga assimilation remains largely unknown in thermophilic microorganisms. In this work, a thermophilic alginolytic community was enriched from coastal sediment, from which an obligate anaerobic and thermophilic bacterial strain, designated Alg1, was isolated. Alg1 shared a 16S rRNA gene identity of 94.6% with Defluviitalea saccharophila LIND6LT2T. Phenotypic, chemotaxonomic, and phylogenetic studies suggested strain Alg1 represented a novel species of the genus Defluviitalea, for which the name Defluviitalea phaphyphila sp. nov. is proposed. Alg1 exhibited an intriguing ability to convert carbohydrates of brown algae, including alginate, laminarin, and mannitol, to ethanol and acetic acid. Three gene clusters participating in this process were predicted to be in the genome, and candidate enzymes were successfully expressed, purified, and characterized. Six alginate lyases were demonstrated to synergistically deconstruct alginate into unsaturated monosaccharide, followed by one uronic acid reductase and two 2-keto-3-deoxy-d-gluconate (KDG) kinases to produce pyruvate. A nonclassical mannitol 1-phosphate dehydrogenase, catalyzing d-mannitol 1-phosphate to fructose 1-phosphate in the presence of NAD+, and one laminarase also were disclosed. This work revealed that a thermophilic brown alga-decomposing system containing numerous novel thermophilic alginate lyases and a unique mannitol 1-phosphate dehydrogenase was adopted by the natural ethanologenic strain Alg1 during the process of evolution in hostile habitats. PMID:26590273

  2. Accumulation of heavy metals (Cu, Cr, Pb and Cd) in freshwater micro algae (Chlorella sp.).

    PubMed

    Kumar, Rajesh M; Frankilin, J; Raj, Samuel Paul

    2013-07-01

    Some selected micro algae were used for the removal of heavy metals from wastewater. In this present investigation, Chlorella sp was studied for accumulation of heavy metals, namely copper, chromium, lead and cadmium. The salts containing heavy metals were dissolved in Blue Green 11 medium at different concentrations in a glass jar of 10 litre capacity each and subsequently they were bubbled with air for 12 days at a temperature of 33 degrees C and light intensity of 2200 lux. The removal rates of heavy metals were recorded for every 4 days during the experimental period. Chlorella sp. removed 37%, 43% and 67% of copper after 4, 8, 12 days respectively. The percentage removal of chromium was 34%, 43% and 50% respectively at 4, 8, and 12 days. Lead removal rates of Chlorella sp were 56% after 4 days, 69% after 8 days and 77% after 12 days. The reduction of cadmium in the culture medium after 12 days was 93%. From the present investigation, it is concluded that heavy metal removal ability of Chlorella sp. can be exploited for metal detoxification and environmental clean up.

  3. Deep Transcriptome Sequencing of Two Green Algae, Chara vulgaris and Chlamydomonas reinhardtii,  Provides No Evidence of Organellar RNA Editing.

    PubMed

    Cahoon, A Bruce; Nauss, John A; Stanley, Conner D; Qureshi, Ali

    2017-02-20

    Nearly all land plants post-transcriptionally modify specific nucleotides within RNAs, a process known as RNA editing. This adaptation allows the correction of deleterious mutations within the asexually reproducing and presumably non-recombinant chloroplast and mitochondrial genomes. There are no reports of RNA editing in any of the green algae so this phenomenon is presumed to have originated in embryophytes either after the invasion of land or in the now extinct algal ancestor of all land plants. This was challenged when a recent in silico screen for RNA edit sites based on genomic sequence homology predicted edit sites in the green alga Chara vulgaris, a multicellular alga found within the Streptophyta clade and one of the closest extant algal relatives of land plants. In this study, the organelle transcriptomes of C. vulgaris and Chlamydomonas reinhardtii were deep sequenced for a comprehensive assessment of RNA editing. Initial analyses based solely on sequence comparisons suggested potential edit sites in both species, but subsequent high-resolution melt analysis, RNase H-dependent PCR (rhPCR), and Sanger sequencing of DNA and complementary DNAs (cDNAs) from each of the putative edit sites revealed them to be either single-nucleotide polymorphisms (SNPs) or spurious deep sequencing results. The lack of RNA editing in these two lineages is consistent with the current hypothesis that RNA editing evolved after embryophytes split from its ancestral algal lineage.

  4. Cultivation of green algae Chlorella sp. in different wastewaters from municipal wastewater treatment plant.

    PubMed

    Wang, Liang; Min, Min; Li, Yecong; Chen, Paul; Chen, Yifeng; Liu, Yuhuan; Wang, Yingkuan; Ruan, Roger

    2010-10-01

    The objective of this study was to evaluate the growth of green algae Chlorella sp. on wastewaters sampled from four different points of the treatment process flow of a local municipal wastewater treatment plant (MWTP) and how well the algal growth removed nitrogen, phosphorus, chemical oxygen demand (COD), and metal ions from the wastewaters. The four wastewaters were wastewater before primary settling (#1 wastewater), wastewater after primary settling (#2 wastewater), wastewater after activated sludge tank (#3 wastewater), and centrate (#4 wastewater), which is the wastewater generated in sludge centrifuge. The average specific growth rates in the exponential period were 0.412, 0.429, 0.343, and 0.948 day(-1) for wastewaters #1, #2, #3, and #4, respectively. The removal rates of NH4-N were 82.4%, 74.7%, and 78.3% for wastewaters #1, #2, and #4, respectively. For #3 wastewater, 62.5% of NO3-N, the major inorganic nitrogen form, was removed with 6.3-fold of NO2-N generated. From wastewaters #1, #2, and #4, 83.2%, 90.6%, and 85.6% phosphorus and 50.9%, 56.5%, and 83.0% COD were removed, respectively. Only 4.7% was removed in #3 wastewater and the COD in #3 wastewater increased slightly after algal growth, probably due to the excretion of small photosynthetic organic molecules by algae. Metal ions, especially Al, Ca, Fe, Mg, and Mn in centrate, were found to be removed very efficiently. The results of this study suggest that growing algae in nutrient-rich centrate offers a new option of applying algal process in MWTP to manage the nutrient load for the aeration tank to which the centrate is returned, serving the dual roles of nutrient reduction and valuable biofuel feedstock production.

  5. Notes on freshwater and terrestrial algae from Ny-Alesund, Svalbard (high Arctic sea area).

    PubMed

    Kim, Gwang Hoon; Klochkova, Tatyana A; Kang, Sung Ho

    2008-07-01

    Field survey of algae and cyanobacteria from terrestrial and freshwater habitats in the vicinity of arctic Ny-Alesund, Svalbard (790N) (high Arctic sea area) was performed in June 2006. Species diversity and abundance were evaluated by using epifluorescence microscopy and culturing methods. In total, 29 taxa in 25 genera were identified, of which Leptolyngbya spp., Trichormus sp. and Chlamydomonas nivalis were abundantly present in almost every sample. In several locations, blooms were formed by species C. nivalis, Scotiellopsis sp., Klebsormidium flaccidum, Zygnema sp., Meridion circulare, Tabellaria fenestrata and Fragilaria sp. Eleven new species from this locality are described.

  6. Sphacelaria lacustris sp. nov. , a freshwater brown alga from Lake Michigan

    SciTech Connect

    Schloesser, R.E.; Blum, J.L.

    1980-06-01

    The growth, reproduction and ultrasturcture of a new freshwater phaeophyte, Sphacelaria lacustris sp. nov., are described. The plant occurs as a minute calcified thallus at 5 to 15 m depth along the western shoreline of Lake Michigan. Both freshly collected and laboratory grown plants show apical growth of erect and basal filaments, intermittent longitudinal divisions in filament segments, vegetative reproduction by propagules, numerous parietal chloroplasts and an absence of pyrenoids, characteristics of Sphacelaria. This material is separated from the only other freshwater species in the genus (S. fluviatilis Jao) at least by differences in longitudinal septation, in branching, in its propagules and in general aspect. Between this plant and marine brown algae there are essential similarities of ultrastructure of cell wall and pores, chloroplasts, mitochondria, nucleus and the production/excretion of physodes.

  7. Formosa algae gen. nov., sp. nov., a novel member of the family Flavobacteriaceae.

    PubMed

    Ivanova, Elena P; Alexeeva, Yulia V; Flavier, Sébastien; Wright, Jonathan P; Zhukova, Natalia V; Gorshkova, Natalia M; Mikhailov, Valery V; Nicolau, Dan V; Christen, Richard

    2004-05-01

    Four light-yellow-pigmented, Gram-negative, short-rod-shaped, non-motile isolates were obtained from enrichment culture during degradation of the thallus of the brown alga Fucus evanescens. The isolates studied were chemo-organotrophic, alkalitolerant and mesophilic. Polar lipids were analysed and phosphatidylethanolamine was the only phospholipid identified. The predominant cellular fatty acids were 15 : 0, i15 : 0, ai15 : 0, i15 : 1 and 15 : 1(n-6). The DNA G+C contents of the four strains were 34.0-34.4 mol%. The level of DNA relatedness of the four isolates was conspecific (88-98 %), indicating that they belong to the same species. The 16S rDNA sequence of strain KMM 3553(T) was determined. Phylogenetic analysis revealed that KMM 3553(T) formed a distinct phyletic line in the phylum Bacteroidetes, class Flavobacteria in the family Flavobacteriaceae and that, phylogenetically, this strain could be placed almost equidistant from the genera Gelidibacter and Psychroserpens (16S rRNA gene sequence similarities of 94 %). On the basis of significant differences in phenotypic and chemotaxonomic characteristics, it is suggested that the isolates represent a novel species in a new genus; the name Formosa algae gen. nov., sp. nov. is proposed. The type strain is KMM 3553(T) (=CIP 107684(T)).

  8. Photosynthetic unit size, carotenoids, and chlorophyll-protein composition of Prochloron sp., a prokaryotic green alga.

    PubMed

    Withers, N W; Alberte, R S; Lewin, R A; Thornber, J P; Britton, G; Goodwin, T W

    1978-05-01

    Six samples of the prokaryotic, unicellular algae Prochloron sp., which occur in association with didemnid ascidians, were collected from various localities in the tropical Pacific Ocean, and their pigments and chlorophyll-protein complexes were identified and characterized. No phycobilin pigments were detected in any of the species. Chlorophylls a and b were present in ratios of a/b = 4.4-6.9. The major carotenoids were beta-carotene (70%) and zeaxanthin (20%). Minor carotenoids of one isolate were identified as echinenone, cryptoxanthin, isocryptoxanthin, mutachrome, and trihydroxy-beta-carotene; no epsilon-ring carotenoids were found in any sample. Except for the absence of glycosidic carotenoids, the overall pigment composition is typical of cyanobacteria. A chlorophyll a/b-protein complex was present in Prochloron; it was electrophoretically and spectrally indistinguishable from the light-harvesting chlorophyll a/b-protein of higher plants and green algae. It accounted for 26% (compared to approximately 50% in green plants) of the total chlorophyll; 17% was associated with a P700-chlorophyll a-protein. The photosynthetic unit size of 240 +/- 10 chlorophylls per P700 in Prochloron was about half that of eukaryotic green plants. A model is proposed for the in vivo organization of chlorophyll in Prochloron.

  9. Formosa undariae sp. nov., isolated from a reservoir containing the brown algae Undaria pinnatifida.

    PubMed

    Park, Sooyeon; Lee, Jung-Sook; Lee, Keun-Chul; Yoon, Jung-Hoon

    2013-11-01

    A strain of Gram-staining-negative, aerobic, non-flagellated, non-gliding and rod-shaped bacteria, designated WS-MY3(T), was isolated from a brown algae reservoir in South Korea. Strain WS-MY3(T) grew optimally at 25 °C, at pH 7.0-8.0 and in the presence of 2.0-3.0 % (w/v) NaCl. Phylogenetic trees based on 16S rRNA gene sequences showed that strain WS-MY3(T) fell within the cluster comprising the type strains of species of the genus Formosa, clustering coherently with the type strains of Formosa agariphila and Formosa algae. It exhibited 16S rRNA gene sequence similarity values of 98.7, 97.9 and 96.8 % to the type strains of F. agariphila, F. algae and Formosa spongicola, respectively. Strain WS-MY3(T) contained MK-6 as the predominant menaquinone and iso-C15 : 0, iso-C16 : 0 3-OH, iso-C15 : 1 G and summed feature 3 (C16 : 1ω7c and/or C16 : 1ω6c) as the major fatty acids. The major polar lipids of strain WS-MY3(T) were phosphatidylethanolamine and two unidentified lipids. The DNA G+C content of strain WS-MY3(T) was 37.3 mol% and its DNA-DNA relatedness values with F. agariphila KCTC 12365(T) and F. algae KCTC 12364(T) were 23 % and 17 %, respectively. The phylogenetic and genetic distinctiveness and differential phenotypic properties revealed that strain WS-MY3(T) is separate from the three recognized species of the genus Formosa. On the basis of the data presented, strain WS-MY3(T) is considered to represent a novel species of the genus Formosa, for which the name Formosa undariae sp. nov. is proposed. The type strain is WS-MY3(T) ( = KCTC 32328(T) = CECT 8286(T)).

  10. Lacinutrix gracilariae sp. nov., a bacterium isolated from the surface of a marine red alga Gracilaria sp.

    PubMed

    Huang, Zhaobin; Li, Guizhen; Lai, Qiliang; Gu, Li; Shao, Zongze

    2015-11-09

    A Gram-negative, aerobic, non-flagellated, rod-shaped bacterium, designated as strain Lxc1T, was isolated from the surface of a marine red alga, Gracilaria sp., which was collected from the coastal regions in Jinjiang, Fujian Province, China. The colony of the strain was orange-yellow, circular and smooth. The 16S rRNA gene of Lxc1T had maximum sequence similarity with Lacinutrix himadriensis E4-9aT (97.1%), followed by L. jangbogonensis PAMC 27137T, L. copepodicola DJ3T, L. algicola AKS293T, and L. mariniflava AKS 432T (similarities <96.4%). Phylogenetic analysis showed strain Lxc1T formed a tight cluster with L. himadriensis E4-9aT and L. copepodicola DJ3T, but represented a novel lineage belonging to the genus Lacinutrix. The predominant fatty acids were iso-C15:1 G (18.3%), iso-C15:0 (16.7%), iso-C17:0-3OH (10.6%), and iso-C15:0-3OH (8.6%). Menaquinone-6 (MK-6) was the only respiratory quinone present. The DNA G+C content of Lxc1T was 31.7 mol%. Combining the results above, it was ascertained that the strain Lxc1T represented a novel species of the genus Lacinutrix, for which the name Lacinutrix gracilariae sp. nov. is proposed. The type strain is Lxc1T (=MCCC 1A01567T=KCTC 42808T).

  11. Feeding characteristics of a golden alga (Poterioochromonas sp.) grazing on toxic cyanobacterium Microcystis aeruginosa.

    PubMed

    Zhang, Xue; Hu, Hong-Ying; Men, Yu-Jie; Yang, Jia; Christoffersen, Kirsten

    2009-07-01

    Microcystis aeruginosa has quickly risen in infamy as one of the most universal and toxic bloom-forming cyanobacteria. Here we presented a species of golden alga (Poterioochromonas sp. strain ZX1), which can feed on toxic M. aeruginosa without any adverse effects from the cyanotoxins. Using flow cytometry, the ingestion and maximal digestion rates were estimated to be 0.2 approximately 1.2 and 0.2 M. aeruginosa cells (ZX1 cell)(-1)h(-1), respectively. M. aeruginosa in densities below 10(7)cells mL(-1) could be grazed down by ZX1, but no significant decrease was observed when the initial density was 3.2 x 10(7)cells mL(-1). ZX1 grazing was a little influenced by the light intensity (0.5 approximately 2500l x) and initial pH of the medium (pH=5.0 approximately 9.5). ZX1 could not survive in continuous darkness for longer than 10 days. The pH value was adjusted to 8 by ZX1 while to 10 by M. aeruginosa. This study may shed light on understanding the ecological interactions between M. aeruginosa and mixotrophic Poterioochromonas sp. in aquatic ecosystems.

  12. Genomics of Volvocine Algae

    PubMed Central

    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

  13. UV radiation-induced accumulation of photoprotective compounds in the green alga Tetraspora sp. CU2551.

    PubMed

    Rastogi, Rajesh P; Incharoensakdi, Aran

    2013-09-01

    The effect of UV radiation on the accumulation of novel mycosporine-like amino acids (MAAs) along with their photoprotective function was investigated in the green alga Tetraspora sp. CU2551. No UV-absorbing compound was detected in this organism growing under normal light condition while two MAAs with absorption maxima at 324 nm and 322 nm were found to be accumulated after UV irradiation. The effects of UV exposure time with different cut-off filter foils namely 295 (PAR + UV-A + UV-B), 320 (PAR + UV-A) and 395 nm (PAR only) were studied on induction of the synthesis of these MAAs. Concentration of MAAs was found to increase with increase in exposure time under UV radiation. Furthermore, the antioxidant and photoprotective action of these MAAs was also investigated. The role of MAAs in diminishing the UV-induced production of ROS in vivo was also demonstrated using the oxidant-sensing probe 2',7'-dichlorodihydrofluorescein diacetate (DCFH-DA) and results obtained supported the results of DPPH free radical scavenging assay. The MAAs also exhibited efficient photoprotective ability on Escherichia coli cells against UV-B stress. Thus, the MAAs in Tetraspora sp. CU2551 may act as efficient antioxidants as well as UV-sunscreen. This is the first report for the UV-induced synthesis and co-accumulation of these MAAs and their photoprotective actions in Tetraspora sp. which is a member of the class Chlorophyceae. Moreover, UV-induced accumulation as well as photoprotective function of these compounds may facilitate this chlorophyte to perform important ecological functions in harsh environmental conditions with high UV-B fluxes in their brightly lit habitats.

  14. Structure and Biological Evaluation of Novel Cytotoxic Sterol Glycosides from the Marine Red Alga Peyssonnelia sp.

    PubMed Central

    Lin, An-Shen; Engel, Sebastian; Smith, Benjamin A.; Fairchild, Craig R.; Aalbersberg, William; Hay, Mark E.; Kubanek, Julia

    2010-01-01

    Bioactivity-guided fractionation of the extract from a Fijian red alga Peyssonnelia sp. led to the isolation of two novel sterol glycosides 19-O-β-d-glucopyranosyl-19-hydroxy-cholest-4-en-3-one (1) and 19-O-β-d-N-acetyl-2-aminoglucopyranosyl-19-hydroxy-cholest-4-en-3-one (2), and two known alkaloids indole-3-carboxaldehyde (3) and 3-(hydroxyacetyl)indole (4). Their structures were characterized by 1D and 2D NMR and mass spectral analysis. The sterol glycosides inhibited cancer cell growth with mean IC50 values (for 11 human cancer cell lines) of 1.63 and 1.41 µM for 1 and 2, respectively. The most sensitive cancer cell lines were MDA-MB-468 (breast) and A549 (lung), with IC50s in of 0.71–0.97 µM for 1 and 2. Modification of the sterol glycoside structures revealed that the α,β-unsaturated ketone at C-3 and oxygenation at C-19 of 1 and 2 are crucial for anticancer activity, whereas the glucosidic group was not essential but contributed to enhanced activity against the most sensitive cell lines. PMID:21036050

  15. Prosthecobacter algae sp. nov., isolated from activated sludge using algal metabolites.

    PubMed

    Lee, Jangho; Park, Banghyo; Woo, Sung-Geun; Lee, Juyoun; Park, Joonhong

    2014-02-01

    A Gram-stain-negative, fusiform-shaped, facultatively anaerobic bacterial strain, designated EBTL04(T), was isolated from activated sludge using algal metabolites and taxonomically characterized through polyphasic investigation. Phylogenetic analysis based on the 16S rRNA gene sequence showed that strain EBTL04(T) belongs to the family Verrucomicrobiaceae, class Verrucomicrobiae, and is closely related to Prosthecobacter dejongeii DSM 12251(T) (98.6 % sequence similarity), Prosthecobacter fusiformis ATCC 25309(T) (97.9 %), Prosthecobacter debontii DSM 14044(T) (97.5%), Prosthecobacter vanneervenii DSM 12252(T) (94.7%) and Prosthecobacter fluviatilis KCTC 22182(T) (93.7%). The G+C content of the genomic DNA of strain EBTL04(T) was 62.7 mol%. The menaquinone MK-6 was detected as the predominant quinone. Strain EBTL04(T) contained phosphatidylethanolamine, phosphatidylglycerol and phosphatidylserine as major polar lipids. A fatty acid profile with C(16 : 1)ω5c, iso-C(14 : 0), C(16 : 0), anteiso-C(15 : 0) and C(14 : 0) as the major components supported the classification of strain EBTL04(T) in the genus Prosthecobacter. Based on several phenotypic, genotypic and chemotaxonomic features, strain EBTL04(T) was clearly differentiated from its phylogenetic neighbours. Therefore, strain EBTL04(T) should be considered to represent a novel species of the genus Prosthecobacter, for which the name Prosthecobacter algae sp. nov. is proposed. The type strain is EBTL04(T) ( = KCTC 23681(T) = JCM 18053(T)).

  16. Molecular mechanisms of the resistance to hydrogen peroxide of enzymes involved in the calvin cycle from halotolerant Chlamydomonas sp. W80.

    PubMed

    Tamoi, M; Kanaboshi, H; Miyasaka, H; Shigeoka, S

    2001-06-15

    cDNA clones encoding NADP(+)-glyceraldehyde-3-phosphate dehydrogenase (NADP(+)-GAPDH) and sedoheptulose-1,7-bisphosphatase (SBPase) were isolated and characterized from halotolerant Chlamydomonas sp. W80 (C. W80) cells. The cDNA clone for NADP(+)-GAPDH encoded 369 amino acid residues, preceded by the chloroplast transit peptide (37 amino acid residues). The cDNA clone for SBPase encoded 351 amino acids with the chloroplast transit peptide. The activities of NADP(+)-GAPDH and SBPase from C. W80 cells were resistant to H(2)O(2) up to 1 mM, as distinct from spinach chloroplastic thiol-modulated enzymes. The illumination to the dark-adapted cells and dithiothreitol treatment to the crude homogenate had little effect on the activities of NADP(+)-GAPDH and SBPase in C. W80. Modeling of the tertiary structures of NADP(+)-GAPDH and SBPase suggests that resistance of the enzymes to H(2)O(2) in C. W80 is due to the different conformational structures in the vicinity of the Cys residues of the chloroplastic enzymes between higher plant and C. W80 cells.

  17. First Report of Pseudobodo sp, a New Pathogen for a Potential Energy-Producing Algae: Chlorella vulgaris Cultures

    PubMed Central

    Zhang, Bangzhou; Yang, Luxi; Zhang, Huajun; Zhang, Jingyan; Li, Yi; Zheng, Wei; Tian, Yun; Liu, Jingwen; Zheng, Tianling

    2014-01-01

    Chlorella vulgaris, is a kind of single-celled green algae, which could serve as a potential source of food and energy because of its photosynthetic efficiency. In our study, a pathogenic organism targeting C. vulgaris was discovered. The algae-lytic activity relates to a fraction from lysates of infected C. vulgaris that was blocked upon filtration through a 3 µm filter. 18S rRNA gene sequence analysis revealed that it shared 99.0% homology with the protist Pseudobodo tremulans. Scanning electron microscope analysis showed that Pseudobodo sp. KD51 cells were approximately 4–5 µm long, biflagellate with an anterior collar around the anterior part of the cell in unstressed feeding cells. Besides the initial host, Pseudobodo sp. KD51 could also kill other algae, indicating its relatively wide predatory spectrum. Heat stability, pH and salinity tolerance experiments were conducted to understand their effects on its predatory activities, and the results showed that Pseudobodo sp. KD51 was heat-sensitive, and pH and salinity tolerant. PMID:24599263

  18. A Comparative biochemical study on two marine endophytes, Bacterium SRCnm and Bacillus sp. JS, Isolated from red sea algae.

    PubMed

    Ahmed, Eman Fadl; Hassan, Hossam Mokhtar; Rateb, Mostafa Ezzat; Abdel-Wahab, Noha; Sameer, Somayah; Aly Taie, Hanan Anwar; Abdel-Hameed, Mohammed Sayed; Hammouda, Ola

    2016-01-01

    Two marine endophytic bacteria were isolated from the Red Sea algae; a red alga; Acanthophora dendroides and the brown alga Sargassum sabrepandum. The isolates were identified based on their 16SrRNA sequences as Bacterium SRCnm and Bacillus sp. JS. The objective of this study was to investigate the potential anti-microbial and antioxidant activities of the extracts of the isolated bacteria grown in different nutrient conditions. Compared to amoxicillin (25μg/disk) and erythromycin (15μg/disk), the extracts of Bacterium SRCn min media II, III, IV and V were potent inhibitors of the gram-positive bacterium Sarcina maxima even at low concentrations. Also, the multidrug resistant Staphylococcus aureus(MRSA) was more sensitive to the metabolites produced in medium (II) of the same endophyte than erythromycin (15μg/disk). A moderate activity of the Bacillus sp. JS extracts of media I and II was obtained against the same pathogen. The total compounds (500ug/ml) of both isolated endophytes showed moderate antioxidant activities (48.9% and 46.1%, respectively). LC/MS analysis of the bacterial extracts was carried out to investigate the likely natural products produced. Cyclo(D-cis-Hyp-L-Leu), dihydrosphingosine and 2-Amino-1,3-hexadecanediol were identified in the fermentation medium of Bacterium SRCnm, whereas cyclo (D-Pro-L-Tyr) and cyclo (L-Leu-L-Pro) were the suggested compounds of Bacillus sp. JS.

  19. Factors determining growth and vertical distribution of planktonic algae in extremely acidic mining lakes (pH 2.7)

    NASA Astrophysics Data System (ADS)

    Bissinger, Vera

    2003-04-01

    In this thesis, I investigated the factors influencing the growth and vertical distribution of planktonic algae in extremely acidic mining lakes (pH 2-3). In the focal study site, Lake 111 (pH 2.7; Lusatia, Germany), the chrysophyte, Ochromonas sp., dominates in the upper water strata and the chlorophyte, Chlamydomonas sp., in the deeper strata, forming a pronounced deep chlorophyll maximum (DCM). Inorganic carbon (IC) limitation influenced the phototrophic growth of Chlamydomonas sp. in the upper water strata. Conversely, in deeper strata, light limited its phototrophic growth. When compared with published data for algae from neutral lakes, Chlamydomonas sp. from Lake 111 exhibited a lower maximum growth rate, an enhanced compensation point and higher dark respiration rates, suggesting higher metabolic costs due to the extreme physico-chemical conditions. The photosynthetic performance of Chlamydomonas sp. decreased in high-light-adapted cells when IC limited. In addition, the minimal phosphorus (P) cell quota was suggestive of a higher P requirement under IC limitation. Subsequently, it was shown that Chlamydomonas sp. was a mixotroph, able to enhance its growth rate by taking up dissolved organic carbon (DOC) via osmotrophy. Therefore, it could survive in deeper water strata where DOC concentrations were higher and light limited. However, neither IC limitation, P availability nor in situ DOC concentrations (bottom-up control) could fully explain the vertical distribution of Chlamydomonas sp. in Lake 111. Conversely, when a novel approach was adopted, the grazing influence of the phagotrophic phototroph, Ochromonas sp., was found to exert top-down control on its prey (Chlamydomonas sp.) reducing prey abundance in the upper water strata. This, coupled with the fact that Chlamydomonas sp. uses DOC for growth, leads to a pronounced accumulation of Chlamydomonas sp. cells at depth; an apparent DCM. Therefore, grazing appears to be the main factor influencing the

  20. The Green Tetrahymena utriculariae n. sp. (Ciliophora, Oligohymenophorea) with Its Endosymbiotic Algae (Micractinium sp.), Living in Traps of a Carnivorous Aquatic Plant.

    PubMed

    Pitsch, Gianna; Adamec, Lubomír; Dirren, Sebastian; Nitsche, Frank; Šimek, Karel; Sirová, Dagmara; Posch, Thomas

    2016-09-10

    The genus Tetrahymena (Ciliophora, Oligohymenophorea) probably represents the best studied ciliate genus. At present, more than forty species have been described. All are colorless, i.e. they do not harbor symbiotic algae, and as aerobes they need at least microaerobic habitats. Here, we present the morphological and molecular description of the first green representative, Tetrahymena utriculariae n. sp., living in symbiosis with endosymbiotic algae identified as Micractinium sp. (Chlorophyta). The full life cycle of the ciliate species is documented, including trophonts and theronts, conjugating cells, resting cysts and dividers. This species has been discovered in an exotic habitat, namely in traps of the carnivorous aquatic plant Utricularia reflexa (originating from Okavango Delta, Botswana). Green ciliates live as commensals of the plant in this anoxic habitat. Ciliates are bacterivorous, however, symbiosis with algae is needed to satisfy cell metabolism but also to gain oxygen from symbionts. When ciliates are cultivated outside their natural habitat under aerobic conditions and fed with saturating bacterial food, they gradually become aposymbiotic. Based on phylogenetic analyses of 18S rRNA and mitochondrial cox1 genes T. utriculariae forms a sister group to Tetrahymena thermophila.

  1. Shewanella algicola sp. nov., a marine bacterium isolated from brown algae.

    PubMed

    Kim, Ji-Young; Yoo, Han-Su; Lee, Dong-Heon; Park, So-Hyun; Kim, Young-Ju; Oh, Duck-Chul

    2016-06-01

    A Gram-stain-negative, aerobic, rod-shaped bacterium motile by means of a single polar flagella, strain ST-6T, was isolated from a brown alga (Sargassum thunbergii) collected in Jeju, Republic of Korea. Strain ST-6T was psychrotolerant, growing at 4-30 °C (optimum 20 °C). Phylogenetic analysis based on 16S rRNA and gyrB gene sequences revealed that strain ST-6T belonged to a distinct lineage in the genus Shewanella. Strain ST-6T was related most closely to Shewanella basaltis J83T, S. gaetbuli TF-27T, S. arctica IT12T, S. vesiculosa M7T and S. aestuarii SC18T, showing 96-97 % and 85-70 % 16S rRNA and gyrB gene sequences similarities, respectively. DNA-DNA relatedness values between strain ST-6T and the type strains of two species of the genus Shewanella were <22.6 %. The major cellular fatty acids (>5 %) were summed feature 3 (comprising C16:1ω7c and/ or iso-C15:0 2-OH), C16:0, iso-C13:0 and C17:1ω8c. The DNA G+C content of strain ST-6Twas 42.4 mol%, and the predominant isoprenoid quinones were menaquinone MK-7 and ubiquinones Q-7 and Q-8. On the basis of its phenotypic properties and phylogenetic distinctiveness, strain ST-6T is considered to represent a novel species of the genus Shewanella, for which the name Shewanella algicola sp. nov. is proposed. The type strain is ST-6T (= KCTC 23253T = JCM 31091T).

  2. Algibacter wandonensis sp. nov., isolated from sediment around a brown algae (Undaria pinnatifida) reservoir.

    PubMed

    Yoon, Jung-Hoon; Park, Sooyeon

    2013-12-01

    A Gram-stain-negative, non-flagellated, rod-shaped bacterial strain able to move by gliding, designated WS-MY22(T), was isolated from sediment around a brown algae reservoir located on Wando in South Korea. It grew optimally at 25 °C, at pH 7.0-8.0 and in the presence of 2.0 % (w/v) NaCl. Phylogenetic trees based on 16S rRNA gene sequences showed that strain WS-MY22(T) clustered coherently with the type strains of Algibacter lectus and Algibacter undariae. It exhibited sequence similarity of 99.4 and 98.9 % to the type strains of A. lectus and A. undariae, respectively, and of 95.1-96.6 % to those of the other species of the genus Algibacter. Strain WS-MY22(T) contained MK-6 as the predominant menaquinone and iso-C15 : 1 G and iso-C17 : 0 3-OH as the major fatty acids. The major polar lipids of strain WS-MY22(T) were phosphatidylethanolamine and two unidentified lipids. The DNA G+C content of strain WS-MY22(T) was 35.8 mol% and its DNA-DNA relatedness with A. lectus KCTC 12103(T) and A. undariae WS-MY9(T) was 31 and 19 %, respectively. The phylogenetic and genetic distinctiveness and differential phenotypic properties revealed that strain WS-MY22(T) is separate from other species of the genus Algibacter. On the basis of the data presented, strain WS-MY22(T) is considered to represent a novel species of the genus Algibacter, for which the name Algibacter wandonensis sp. nov. is proposed. The type strain is WS-MY22(T) ( = KCTC 32381(T) = CECT 8301(T)).

  3. Winogradskyella eckloniae sp. nov., a marine bacterium isolated from the brown alga Ecklonia cava.

    PubMed

    Kim, Ji-Young; Park, So-Hyun; Seo, Ga-Young; Kim, Young-Ju; Oh, Duck-Chul

    2015-09-01

    A novel bacterial strain, designated EC29(T), was isolated from the brown alga Ecklonia cava collected on Jeju Island, Republic of Korea. Cells of strain EC29(T) were Gram-stain-negative, aerobic, rod-shaped and motile by gliding. Growth was observed at 10-30 °C (optimum, 20-25 °C), at pH 6.0-9.5 (optimum, pH 7.5) and in the presence of 1-5% (w/v) NaCl. Phylogenetic analyses based on the 16S rRNA gene sequence revealed that the strain belonged to the genus Winogradskyella. Strain EC29(T) exhibited the highest 16S rRNA gene sequence similarities, of 96.5-97.8%, to the type strains of Winogradskyella pulchriflava EM106(T), Winogradskyella echinorum KMM 6211(T) and Winogradskyella ulvae KMM 6390(T). Strain EC29(T) exhibited < 27% DNA-DNA relatedness with Winogradskyella pulchriflava EM106(T) and Winogradskyella echinorum KMM 6211(T). The predominant fatty acids of strain EC29(T) were iso-C15 : 0, iso-C15 : 1 G, C15 : 0, iso-C17 : 0 3-OH, iso-C15 : 0 3-OH and anteiso-C15 : 0. The DNA G+C content was 31.1 mol% and the major respiratory quinone was menaquinone-6 (MK-6). Based on a polyphasic study, strain EC29(T) is considered to represent a novel species of the genus Winogradskyella, for which the name Winogradskyella eckloniae sp. nov. is proposed. The type strain is EC29(T) ( = KCTC 32172(T) = JCM 18703(T)).

  4. Sulfitobacter porphyrae sp. nov., isolated from the red alga Porphyra yezoensis.

    PubMed

    Fukui, Youhei; Abe, Mahiko; Kobayashi, Masahiro; Shimada, Yushi; Saito, Hiroaki; Oikawa, Hiroshi; Yano, Yutaka; Satomi, Masataka

    2014-02-01

    Gram-stain-negative, aerobic, halophilic bacteria, designated SCM-1(T), LCM10-1 and CTBL-B-147, were isolated from modified half-strength SWM-III medium, PES medium and thalli after laboratory cultivation of a red alga, Porphyra yezoensis. Phylogenetic analysis of 16S rRNA gene sequences indicated that the new isolates were affiliated to the genus Sulfitobacter of the class Alphaproteobacteria, and the 16S rRNA gene sequence similarity of the new isolates with the closest related species, Sulfitobacter mediterraneus CH-B427(T), was 98.8%. The DNA G+C contents of the new isolates were in the range of 61.4-62.3 mol%. DNA-DNA relatedness values of strain SCM-1(T) with other type strains of the genus Sulfitobacter were less than 15.9%. The new isolates contained Q-10 as the predominant ubiquinone, phosphatidylcholine, phosphatidylglycerol, an unidentified amino lipid and an unidentified lipid as the main polar lipids, and C(18 : 1)ω7c, C(19 : 1)ω7c and C(16 : 0) as the major fatty acids (>10% of the total). Strain SCM-1(T) could be differentiated from Sulfitobacter mediterraneus JCM 21792(T) by 35 morphological and phenotypic characteristics. On the basis of the phylogenetic, genetic and phenotypic properties of the new isolates, the name Sulfitobacter porphyrae sp. nov. is proposed, with strain SCM-1(T) ( = LMG 27110(T) = NBRC 109054(T)) as the type strain.

  5. Flavobacterium jejuensis sp. nov., isolated from marine brown alga Ecklonia cava.

    PubMed

    Park, So-Hyun; Kim, Ji-Young; Kim, Young-Ju; Heo, Moon-Soo

    2015-11-01

    A bacterial strain, designated EC11(T) was isolated from brown alga Ecklonia cava collected from Jeju Island, Korea. EC11(T) was identified as a Gram-negative, rod-shaped and yellow-pigmented bacterial strain. The strain EC11(T) grew over a temperature range of 10 °C to 30 °C (optimally at 25 °C), and a pH range of 6.0-10.5 (optimally at pH 7.5). Phylogenetic analysis based on 16S rRNA gene sequences showed that strain EC11(T) belongs to the genus Flavobacterium. Strain EC11(T) shared close similarity with Flavobacterium jumunjinense HME7102(T) (96.4%), Flavobacterium dongtanense LW30(T) (95.8%), Flavobacterium haoranii LQY-7(T) (95.3%), and Flavobacterium urocaniciphilum (95.1%). The major fatty acids (> 5%) were iso-C17:0 3-OH (22.4%), iso-C15:0 3-OH (19.0%), C15:0 (12.4%), summed feature 3 (comprising C16:1 ω7c/ C16:1 ω6c; 9.78%), iso-C15:1 G (9.6%), and iso-C16:0 3-OH (9.0%). The DNA G+C content was 28.1 mol% and the strain contained MK-6 as the predominant menaquinone. The major polar lipids were phosphatidylethanolamine, two unknown aminolipids and three unknown polar lipids. Based on phenotypic, chemotaxonomic and phylogenetic analysis, strain EC11T represents a novel species of the Flavobacterium genus, for which the name Flavobacterium jejuensis sp. nov. is proposed. The type strain of F. jejuensis is EC11(T) (=KCTC 42149(T) = JCM 30735(T)).

  6. The cell-wall glycoproteins of the green alga Scenedesmus obliquus. The predominant cell-wall polypeptide of Scenedesmus obliquus is related to the cell-wall glycoprotein gp3 of Chlamydomonas reinhardtii.

    PubMed

    Voigt, Jürgen; Stolarczyk, Adam; Zych, Maria; Malec, Przemysław; Burczyk, Jan

    2014-02-01

    The green alga Scenedesmus obliquus contains a multilayered cell wall, ultrastructurally similar to that of Chlamydomonas reinhardtii, although its proportion of hydroxyproline is considerably lower. Therefore, we have investigated the polypeptide composition of the insoluble and the chaotrope-soluble wall fractions of S. obliquus. The polypeptide pattern of the chaotrope-soluble wall fraction was strongly modified by chemical deglycosylation with anhydrous hydrogen fluoride (HF) in pyridine indicating that most of these polypeptides are glycosylated. Polypeptide constituents of the chaotrope-soluble cell-wall fraction with apparent molecular masses of 240, 270, 265, and 135 kDa cross-reacted with a polyclonal antibody raised against the 100 kDa deglycosylation product of the C. reinhardtii cell-wall glycoprotein GP3B. Chemical deglycosylation of the chaotrope-soluble wall fraction resulted in a 135 kDa major polypeptide and a 106 kDa minor component reacting with the same antibody. This antibody recognized specific peptide epitopes of GP3B. When the insoluble wall fraction of S. obliquus was treated with anhydrous HF/pyridine, three polypeptides with apparent molecular masses of 144, 135, and 65 kDa were solubilized, which also occured in the deglycosylated chaotrope-soluble wall fraction. These findings indicate that theses glycoproteins are cross-linked to the insoluble wall fraction via HF-sensitive bonds.

  7. Mychonastes desiccatus Brown sp. nova (Chlorococcales, Chlorophyta)--an intertidal alga forming achlorophyllous desiccation-resistant cysts

    NASA Technical Reports Server (NTRS)

    Margulis, L.; Hinkle, G.; McKhann, H.; Moynihan, B.

    1988-01-01

    An intertidal Chlorella-like alga Mychonastes desiccatus Brown sp. nova, capable of forming achlorophyllous desiccation-resistant cysts, has been grown in unialgal culture. This small alga was first isolated from a dried sample of a well-studied microbial mat. The mat, located at North Pond, Laguna Figueroa, San Quintin, Baja California, Mexico, is a vertically-stratified microbial community which forms laminated sediments. Morphology, pigment composition and G+C content are within the range typical for the genus Chlorella s. 1. Unlike other chlorellae, however, upon desiccation M. desiccatus forms an achlorophyllous, lipid-filled cyst (thick-walled resting stage) in which no plastid is evident. Rewetting leads to chloroplast differentiation, excystment and recovery of the fully green alga. During desiccation, sporopollenin is deposited within a thickening cell wall. Encystment cannot be induced by growth in the dark. The formation of desiccation-induced cysts allows the alga to survive frequent and intermittent periods of dryness. These chlorellae tolerate wide ranges of acidity and temperature; they both grow and form cysts in media in which sodium ions are replaced with potassium. Although the cysts tolerate crystalline salts, the cell grow optimally in concentrations corresponding from three-quarters to full-strength seawater.

  8. Mychonastes desiccatus Brown sp. nova (Chlorococcales, Chlorophyta)--an intertidal alga forming achlorophyllous desiccation-resistant cysts.

    PubMed

    Margulis, L; Hinkle, G; McKhann, H; Moynihan, B

    1988-09-01

    An intertidal Chlorella-like alga Mychonastes desiccatus Brown sp. nova, capable of forming achlorophyllous desiccation-resistant cysts, has been grown in unialgal culture. This small alga was first isolated from a dried sample of a well-studied microbial mat. The mat, located at North Pond, Laguna Figueroa, San Quintin, Baja California, Mexico, is a vertically-stratified microbial community which forms laminated sediments. Morphology, pigment composition and G+C content are within the range typical for the genus Chlorella s. 1. Unlike other chlorellae, however, upon desiccation M. desiccatus forms an achlorophyllous, lipid-filled cyst (thick-walled resting stage) in which no plastid is evident. Rewetting leads to chloroplast differentiation, excystment and recovery of the fully green alga. During desiccation, sporopollenin is deposited within a thickening cell wall. Encystment cannot be induced by growth in the dark. The formation of desiccation-induced cysts allows the alga to survive frequent and intermittent periods of dryness. These chlorellae tolerate wide ranges of acidity and temperature; they both grow and form cysts in media in which sodium ions are replaced with potassium. Although the cysts tolerate crystalline salts, the cell grow optimally in concentrations corresponding from three-quarters to full-strength seawater.

  9. Tetraflagellochloris mauritanica gen. et sp. nov. (Chlorophyceae), a New Flagellated Alga from the Mauritanian Desert: Morphology, Ultrastructure, and Phylogenetic Framing.

    PubMed

    Barsanti, Laura; Frassanito, Anna Maria; Passarelli, Vincenzo; Evangelista, Valtere; Etebari, Maryam; Paccagnini, Eugenio; Lupetti, Pietro; Lenzi, Paola; Verni, Franco; Gualtieri, Paolo

    2013-02-01

    Morphological, ultrastructural, and molecular-sequence data were used to assess the phylogenetic position of a tetraflagellate green alga isolated from soil samples of a saline dry basin near F'derick, Mauritania. This alga can grow as individual cells or form non-coenobial colonies of up to 12 individuals. It has a parietal chloroplast with an embedded pyrenoid covered by a starch sheath and traversed by single parallel thylakoids, and an eyespot located in a parietal position opposite to the flagellar insertion. Lipid vacuoles are present in the cytoplasm. Microspectroscopy indicated the presence of chlorophylls a and b, with lutein as the major carotenoid in the chloroplast, while the eyespot spectrum has a shape typical of green-algal eyespots. The cell has four flagella, two of them long and two considerably shorter. Sequence data from the 18S rRNA gene and ITS2 were obtained and compared with published sequences for green algae. Results from morphological and ultrastructural examinations and sequence analysis support the placement of this alga in the Chlorophyceae, as Tetraflagellochloris mauritanica L. Barsanti et A. Barsanti, gen. et sp. nov.

  10. Separate origins of ice-binding proteins in antarctic chlamydomonas species.

    PubMed

    Raymond, James A; Morgan-Kiss, Rachael

    2013-01-01

    The green alga Chlamydomonas raudensis is an important primary producer in a number of ice-covered lakes and ponds in Antarctica. A C. raudensis isolate (UWO241) from Lake Bonney in the McMurdo Dry Valleys, like many other Antarctic algae, was found to secrete ice-binding proteins (IBPs), which appear to be essential for survival in icy environments. The IBPs of several Antarctic algae (diatoms, a prymesiophyte, and a prasinophyte) are similar to each other (here designated as type I IBPs) and have been proposed to have bacterial origins. Other IBPs (type II IBPs) that bear no resemblance to type I IBPs, have been found in the Antarctic Chlamydomonas sp. CCMP681, a putative snow alga, raising the possibility that chlamydomonad IBPs developed separately from the IBPs of other algae. To test this idea, we obtained the IBP sequences of C. raudensis UWO241 by sequencing the transcriptome. A large number of transcripts revealed no sequences resembling type II IBPs. Instead, many isoforms resembling type I IBPs were found, and these most closely matched a hypothetical protein from the bacterium Stigmatella aurantiaca. The sequences were confirmed to encode IBPs by the activity of a recombinant protein and by the matching of predicted and observed isoelectric points and molecular weights. Furthermore, a mesophilic sister species, C. raudensis SAG49.72, showed no ice-binding activity or PCR products from UWO241 IBP primers. These results confirm that algal IBPs are required for survival in icy habitats and demonstrate that they have diverse origins that are unrelated to the taxonomic positions of the algae. Last, we show that the C. raudensis UWO241 IBPs can change the structure of ice in a way that could increase the survivability of cells trapped in the ice.

  11. Cultivation of Acidophilic Algae Galdieria sulphuraria and Pseudochlorella sp. YKT1 in Media Derived from Acidic Hot Springs.

    PubMed

    Hirooka, Shunsuke; Miyagishima, Shin-Ya

    2016-01-01

    Microalgae possess a high potential for producing pigments, antioxidants, and lipophilic compounds for industrial applications. However, the cultivation of microalgae comes at a high cost. To reduce the cost, changes from a closed bioreactor to open pond system and from a synthetic medium to environmental or wastewater-based medium are being sought. However, the use of open pond systems is currently limited because of contamination by undesirable organisms. To overcome this issue, one strategy is to combine acidophilic algae and acidic drainage in which other organisms are unable to thrive. Here, we tested waters from sulfuric acidic hot springs (Tamagawa, pH 1.15 and Tsukahara, pH 1.14) in Japan for the cultivation of the red alga Galdieria sulphuraria 074G and the green alga Pseudochlorella sp. YKT1. Both of these spring waters are rich in phosphate (0.043 and 0.145 mM, respectively) compared to other environmental freshwater sources. Neither alga grew in the spring water but they grew very well when the waters were supplemented with an inorganic nitrogen source. The algal yields were ∼2.73 g dry weight/L for G. sulphuraria and ∼2.49 g dry weight/L for P. sp. YKT1, which were comparable to those in an autotrophic synthetic medium. P. sp. YKT1 grew in the spring waters supplemented either of NH4(+), NO3(-) or urea, while G. sulphuraria grew only when NH4(+) was supplemented. For P. sp. YKT1, the spring water was adjusted to pH 2.0, while for G. sulphuraria, no pH adjustment was required. In both cases, no additional pH-buffering compound was required. The phycocyanin of the thermophilic G. sulphuraria is known to be more thermostable than that from the Spirulina platensis currently used in phycocyanin production for commercial use. The phycocyanin content in G. sulphuraria in the Tsukahara water supplemented with NH4(+) was 107.42 ± 1.81 μg/mg dry weight, which is comparable to the level in S. platensis (148.3 μg/mg dry weight). P. sp. YKT1 cells in the

  12. Cultivation of Acidophilic Algae Galdieria sulphuraria and Pseudochlorella sp. YKT1 in Media Derived from Acidic Hot Springs

    PubMed Central

    Hirooka, Shunsuke; Miyagishima, Shin-ya

    2016-01-01

    Microalgae possess a high potential for producing pigments, antioxidants, and lipophilic compounds for industrial applications. However, the cultivation of microalgae comes at a high cost. To reduce the cost, changes from a closed bioreactor to open pond system and from a synthetic medium to environmental or wastewater-based medium are being sought. However, the use of open pond systems is currently limited because of contamination by undesirable organisms. To overcome this issue, one strategy is to combine acidophilic algae and acidic drainage in which other organisms are unable to thrive. Here, we tested waters from sulfuric acidic hot springs (Tamagawa, pH 1.15 and Tsukahara, pH 1.14) in Japan for the cultivation of the red alga Galdieria sulphuraria 074G and the green alga Pseudochlorella sp. YKT1. Both of these spring waters are rich in phosphate (0.043 and 0.145 mM, respectively) compared to other environmental freshwater sources. Neither alga grew in the spring water but they grew very well when the waters were supplemented with an inorganic nitrogen source. The algal yields were ∼2.73 g dry weight/L for G. sulphuraria and ∼2.49 g dry weight/L for P. sp. YKT1, which were comparable to those in an autotrophic synthetic medium. P. sp. YKT1 grew in the spring waters supplemented either of NH4+, NO3- or urea, while G. sulphuraria grew only when NH4+ was supplemented. For P. sp. YKT1, the spring water was adjusted to pH 2.0, while for G. sulphuraria, no pH adjustment was required. In both cases, no additional pH-buffering compound was required. The phycocyanin of the thermophilic G. sulphuraria is known to be more thermostable than that from the Spirulina platensis currently used in phycocyanin production for commercial use. The phycocyanin content in G. sulphuraria in the Tsukahara water supplemented with NH4+ was 107.42 ± 1.81 μg/mg dry weight, which is comparable to the level in S. platensis (148.3 μg/mg dry weight). P. sp. YKT1 cells in the Tamagawa

  13. Development of the light-harvesting chlorophyll antenna in the green alga Chlamydomonas reinhardtii is regulated by the novel Tla1 gene.

    PubMed

    Tetali, Sarada D; Mitra, Mautusi; Melis, Anastasios

    2007-03-01

    The Chlamydomonas reinhardtii tla1 (truncated light-harvesting chlorophyll antenna size) mutant was generated upon DNA insertional mutagenesis and shown to specifically possess a smaller than wild type (WT) chlorophyll antenna size in both photosystems. Molecular and genetic analysis revealed that the exogenous plasmid DNA was inserted at the end of the 5' UTR and just prior to the ATG start codon of a hitherto unknown nuclear gene (termed Tla1), which encodes a protein of 213 amino acids. The Tla1 gene in the mutant is transcribed with a new 5' UTR sequence, derived from the 3' end of the transforming plasmid. This replacement of the native 5' UTR and promoter regions resulted in enhanced transcription of the tla1 gene in the mutant but inhibition in the translation of the respective tla1 mRNA. Transformation of the tla1 mutant with WT Tla1 genomic DNA successfully rescued the mutant. These results are evidence that polymorphism in the 5' UTR of the Tla1 transcripts resulted in the tla1 phenotype and that expression of the Tla1 gene is a prerequisite for the development/assembly of the Chl antenna in C. reinhardtii. A blast search with the Tla1 deduced amino acid sequence

  14. Biochemical and Structural Studies of the Large Ycf4-Photosystem I Assembly Complex of the Green Alga Chlamydomonas reinhardtii[W

    PubMed Central

    Ozawa, Shin-ichiro; Nield, Jon; Terao, Akihiro; Stauber, Einar J.; Hippler, Michael; Koike, Hiroyuki; Rochaix, Jean-David; Takahashi, Yuichiro

    2009-01-01

    Ycf4 is a thylakoid protein essential for the accumulation of photosystem I (PSI) in Chlamydomonas reinhardtii. Here, a tandem affinity purification tagged Ycf4 was used to purify a stable Ycf4-containing complex of >1500 kD. This complex also contained the opsin-related COP2 and the PSI subunits PsaA, PsaB, PsaC, PsaD, PsaE, and PsaF, as identified by mass spectrometry (liquid chromatography–tandem mass spectrometry) and immunoblotting. Almost all Ycf4 and COP2 in wild-type cells copurified by sucrose gradient ultracentrifugation and subsequent ion exchange column chromatography, indicating the intimate and exclusive association of Ycf4 and COP2. Electron microscopy revealed that the largest structures in the purified preparation measure 285 × 185 Å; these particles may represent several large oligomeric states. Pulse-chase protein labeling revealed that the PSI polypeptides associated with the Ycf4-containing complex are newly synthesized and partially assembled as a pigment-containing subcomplex. These results indicate that the Ycf4 complex may act as a scaffold for PSI assembly. A decrease in COP2 to 10% of wild-type levels by RNA interference increased the salt sensitivity of the Ycf4 complex stability but did not affect the accumulation of PSI, suggesting that COP2 is not essential for PSI assembly. PMID:19700633

  15. Biosorption of copper, cobalt and nickel by marine brown alga Sargassum sp. in fixed-bed column.

    PubMed

    Esmaeili, Akbar; Soufi, Samira; Rustaiyan, Abdolhossein; Safaiyan, Shila; Mirian, Simin; Fallahe, Gila; Moazami, Nasrin

    2007-11-01

    The biosorption of copper, cobalt and nickel by marine brown alga Sargassum sp. were investigated in a fixed-bed column (temperature = 30 degrees C; different pH). Langmuir and Freundlich sorption models were used to represent the equilibrium data. The maximum Cu2+ uptake was obtained at pH 4 and the optimum Co2+ and Ni2+ uptake were at pH 7. Different dosage of biosorbent did not have an effect on the results, but the 3.5 and 5 g of biosorbent were shown higher uptake. The metal removal rates were rapid, with about 80% of the total adsorption tacking place within 40 min.

  16. Winogradskyella undariae sp. nov., a member of the family Flavobacteriaceae isolated from a brown algae reservoir.

    PubMed

    Park, Sooyeon; Yoon, Jung-Hoon

    2013-11-01

    A novel bacterial strain, designated WS-MY5(T), capable of degrading a variety of polysaccharides was isolated from a brown algae (Undaria pinnatifida) reservoir at Wando in the South Sea, South Korea. Strain WS-MY5(T) was found to grow optimally at 30 °C, at pH 7.0-7.5 and in the presence of 2 % (w/v) NaCl. A neighbour-joining phylogenetic tree based on 16S rRNA gene sequences showed that strain WS-MY5(T) falls within the clade comprising Winogradskyella species, clustering with the type strains of Winogradskyella pacifica, Winogradskyella arenosi, Winogradskyella rapida and Winogradskyella thalassocola, with which it exhibited 16S rRNA gene sequence similarity values of 97.3-98.8 %. It exhibited sequence similarity values of 93.0-96.2 % to the type strains of the other recognized Winogradskyella species. Strain WS-MY5(T) was found to contain MK-6 as the predominant menaquinone and anteiso-C15:0, iso-C15:0, iso-C15:0 3-OH, iso-C17:0 3-OH and iso-C15:1 G as the major fatty acids. The major polar lipids of strain WS-MY5(T) were identified as phosphatidylethanolamine, two unidentified lipids and two unidentified aminolipids. The DNA G+C content of strain WS-MY5(T) was determined to be 33.2 mol% and its DNA-DNA relatedness values with the type strains of W. pacifica, W. arenosi, W. rapida and W. thalassocola were in the range 16-28 %. Differential phenotypic properties, together with its phylogenetic and genetic distinctiveness, enabled strain WS-MY5(T) to be differentiated from the recognized Winogradskyella species. On the basis of the data presented here, strain WS-MY5(T) is considered to represent a novel species of the genus Winogradskyella, for which the name Winogradskyella undariae sp. nov. is proposed. The type strain is WS-MY5(T) (=KCTC 32261(T)=CCUG 63832(T)).

  17. Leuconostoc miyukkimchii sp. nov., isolated from brown algae (Undaria pinnatifida) kimchi.

    PubMed

    Lee, Seung Hyeon; Park, Moon Su; Jung, Ji Young; Jeon, Che Ok

    2012-05-01

    A Gram-staining-positive, non-motile and non-spore-forming lactic acid bacterium, designated strain M2(T), was isolated from fermented brown algae (Undaria pinnatifida) kimchi in South Korea. Cells of the isolate were facultatively anaerobic ovoids and showed catalase- and oxidase-negative reactions. Growth of strain M2(T) was observed at 4-35 °C and at pH 5.0-9.0. The G+C content of the genomic DNA was 42.5 mol%. Phylogenetic analysis based on 16S rRNA gene sequences indicated that strain M2(T) belonged to the genus Leuconostoc and was most closely related to Leuconostoc inhae IH003(T), Leuconostoc kimchii IH25(T), Leuconostoc gasicomitatum LMG 18811(T), Leuconostoc gelidum DSM 5578(T), Leuconostoc palmae TMW2.694(T) and Leuconostoc holzapfelii BFE 7000(T) with 98.9 %, 98.8 %, 98.8 %, 98.7 %, 98.5 % and 98.2 % sequence similarity, respectively. DNA-DNA hybridization values between strain M2(T) and Leuconostoc inhae KACC 12281(T), Leuconostoc kimchii IH25(T), Leuconostoc gelidum KACC 12256(T), Leuconostoc gasicomitatum KACC 13854(T), Leuconostoc palmae DSM 21144(T) and Leuconostoc holzapfelii DSM 21478(T) were 13.8±3.2 %, 14.3±3.4 %, 9.9±1.0 %, 13.2±0.8 %, 22.4±4.9 % and 16.2±4.6 %, respectively, which allowed differentiation of strain M2(T) from the closely related species of the genus Leuconostoc. On the basis of phenotypic and molecular properties, strain M2(T) represents a novel species in the genus Leuconostoc, for which the name Leuconostoc miyukkimchii sp. nov. is proposed. The type strain is M2(T) ( = KACC 15353(T)  = JCM 17445(T)).

  18. CHLAMYDOMONAS FLAGELLA

    PubMed Central

    Witman, G. B.; Carlson, K.; Rosenbaum, Joel L.

    1972-01-01

    Quantitative ultrastructural analysis and quantitative gel electrophoresis of preparations of selectively solubilized Chlamydomonas outer doublets indicated that tubulins 1 and 2 were present in both the A tubule and the B tubule, and that only tubulin 1 was present in the three protofilaments which form the wall ("partition") between the lumens of the A and B tubules. The data suggested that the remaining protofilaments of the outer doublet were grouped together in pairs containing the same type of tubulin, pairs containing tubulin 1 alternating with pairs containing tubulin 2. These findings were used to construct models for the arrangement of the two tubulins in the outer doublet. Further analysis by isoelectric focusing resolved tubulins 1 and 2 into at least five bands. PMID:5044758

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

    PubMed

    Pugliara, Alessandro; Makasheva, Kremena; Despax, Bernard; Bayle, Maxime; Carles, Robert; Benzo, Patrizio; BenAssayag, Gérard; Pécassou, Béatrice; Sancho, Maria Carmen; Navarro, Enrique; Echegoyen, Yolanda; Bonafos, Caroline

    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 <20nm) 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 7nm) 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 20h of immersion in buffered water was measured by inductively coupled plasma mass spectrometry and ranges between 0.02 and 0.49μM. 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.

  20. A brief introduction to the model microswimmer Chlamydomonas reinhardtii

    NASA Astrophysics Data System (ADS)

    Jeanneret, Raphaël; Contino, Matteo; Polin, Marco

    2016-11-01

    The unicellular biflagellate green alga Chlamydomonas reinhardtii has been an important model system in biology for decades, and in recent years it has started to attract growing attention also within the biophysics community. Here we provide a concise review of some of the aspects of Chlamydomonas biology and biophysics most immediately relevant to physicists that might be interested in starting to work with this versatile microorganism.

  1. Acclimation of Antarctic Chlamydomonas to the sea-ice environment: a transcriptomic analysis.

    PubMed

    Liu, Chenlin; Wang, Xiuliang; Wang, Xingna; Sun, Chengjun

    2016-07-01

    The Antarctic green alga Chlamydomonas sp. ICE-L was isolated from sea ice. As a psychrophilic microalga, it can tolerate the environmental stress in the sea-ice brine, such as freezing temperature and high salinity. We performed a transcriptome analysis to identify freezing stress responding genes and explore the extreme environmental acclimation-related strategies. Here, we show that many genes in ICE-L transcriptome that encoding PUFA synthesis enzymes, molecular chaperon proteins, and cell membrane transport proteins have high similarity to the gens from Antarctic bacteria. These ICE-L genes are supposed to be acquired through horizontal gene transfer from its symbiotic microbes in the sea-ice brine. The presence of these genes in both sea-ice microalgae and bacteria indicated the biological processes they involved in are possibly contributing to ICE-L success in sea ice. In addition, the biological pathways were compared between ICE-L and its closely related sister species, Chlamydomonas reinhardtii and Volvox carteri. In ICE-L transcripome, many sequences homologous to the plant or bacteria proteins in the post-transcriptional, post-translational modification, and signal-transduction KEGG pathways, are absent in the nonpsychrophilic green algae. These complex structural components might imply enhanced stress adaptation capacity. At last, differential gene expression analysis at the transcriptome level of ICE-L indicated that genes that associated with post-translational modification, lipid metabolism, and nitrogen metabolism are responding to the freezing treatment. In conclusion, the transcriptome of Chlamydomonas sp. ICE-L is very useful for exploring the mutualistic interaction between microalgae and bacteria in sea ice; and discovering the specific genes and metabolism pathways responding to the freezing acclimation in psychrophilic microalgae.

  2. First record of the insect pathogenic alga Helicosporidium sp. (Chlorophyta: Trebouxiophyceae) infection in larvae and pupae of Rhizophagusgrandis Gyll. (Coleoptera, Rhizophaginae) from Turkey.

    PubMed

    Yaman, Mustafa; Radek, Renate; Aydin, Ciçek; Tosun, Onur; Ertürk, Omer

    2009-10-01

    The predator beetle Rhizophagus grandis Gyll. (Coleoptera, Rhizophaginae) is one of the most important biological control agents, mass-bred and used to suppress populations of an important pest: the great spruce bark beetle, Dendroctonus micans. The achlorophyllous alga Helicosporidium sp. was first discovered in the pest. Later it was also found in the predator, but only in the adults. In this study, the pathogenic alga Helicosporidium sp. was discovered in larvae and early pupae of R. grandis for the first time. The morphological characteristics of the pathogenic alga were revealed by light and electron microscopy. Infection rates of Helicosporidium sp. in the larvae and pupae of R. grandis were 23.5% and 6.25%, respectively.

  3. Hydrogen photoproduction in green algae Chlamydomonas reinhardtii sustainable over 2 weeks with the original cell culture without supply of fresh cells nor exchange of the whole culture medium.

    PubMed

    Yagi, Takafumi; Yamashita, Kyohei; Okada, Norihide; Isono, Takumi; Momose, Daisuke; Mineki, Shigeru; Tokunaga, Eiji

    2016-07-01

    Unicellular green algae Chlamydomonas reinhardtii are known to make hydrogen photoproduction under the anaerobic condition with water molecules as the hydrogen source. Since the hydrogen photoproduction occurs for a cell to circumvent crisis of its survival, it is only temporary. It is a challenge to realize persistent hydrogen production because the cells must withstand stressful conditions to survive with alternation of generations in the cell culture. In this paper, we have found a simple and cost-effective method to sustain the hydrogen production over 14 days in the original culture, without supply of fresh cells nor exchange of the culture medium. This is achieved for the cells under hydrogen production in a sulfur-deprived culture solution on the {anaerobic, intense light} condition in a desiccator, by periodically providing a short period of the recovery time (2 h) with a small amount of TAP(+S) supplied outside of the desiccator. As this operation is repeated, the response time of transition into hydrogen production (preparation time) is shortened and the rate of hydrogen production (build up time) is increased. The optimum states of these properties favorable to the hydrogen production are attained in a few days and stably sustained for more than 10 days. Since generations are alternated during this consecutive hydrogen production experiment, it is suggested that the improved hydrogen production properties are inherited to next generations without genetic mutation. The properties are reset only when the cells are placed on the {sulfur-sufficient, aerobic, moderate light} conditions for a long time (more than 1 day at least).

  4. Polyclonal antibodies against the TLA1 protein also recognize with high specificity the D2 reaction center protein of PSII in the green alga Chlamydomonas reinhardtii.

    PubMed

    Mitra, Mautusi; Dewez, David; García-Cerdán, Jose Gines; Melis, Anastasios

    2012-04-01

    The Chlamydomonas reinhardtii DNA-insertional transformant truncated light-harvesting antenna 1 (tla1) mutant, helped identify the novel TLA1 gene (GenBank Accession # AF534570-71) as an important genetic determinant in the chlorophyll antenna size of photosynthesis. Down-regulation in the amount of the TLA1 23 kDa protein in the cell resulted in smaller chlorophyll antenna size for both photosystems (in Tetali et al. Planta 225:813-829, 2007). Specific polyclonal antibodies, raised against the recombinant TLA1 protein, showed a cross-reaction with the predicted 23 kDa TLA1 protein in C. reinhardtii protein extracts, but also showed a strong cross-reaction with a protein band migrating to 28.5 kDa. Questions of polymorphism, or posttranslational modification of the TLA1 protein were raised as a result of the unexpected 28.5 kDa cross-reaction. Work in this paper aimed to elucidate the nature of the unexpected 28.5 kDa cross-reaction, as this was deemed to be important in terms of the functional role of the TLA1 protein in the regulation of the chlorophyll antenna size of photosynthesis. Immuno-precipitation of the 28.5 kDa protein, followed by LC-mass spectrometry, showed amino acid sequences ascribed to the psbD/D2 reaction center protein of PSII. The common antigenic determinant between TLA1 and D2 was shown to be a stretch of nine conserved amino acids V-F-L(V)LP-GNAL in the C-terminus of the two proteins, constituting a high antigenicity "GNAL" domain. Antibodies raised against the TLA1 protein containing this domain recognized both the TLA1 and the D2 protein. Conversely, antibodies raised against the TLA1 protein minus the GNAL domain specifically recognized the 23 kDa TLA1 protein and failed to recognize the 28.5 kDa D2 protein. D2 antibodies raised against an oligopeptide containing this domain also cross-reacted with the TLA1 protein. It is concluded that the 28.5 kDa cross-reaction of C. reinhardtii protein extracts with antiTLA1 antibodies is due to

  5. The ice nucleation activity of extremophilic algae.

    PubMed

    Kviderova, Jana; Hajek, Josef; Worland, Roger M

    2013-01-01

    Differences in the level of cold acclimation and cryoprotection estimated as ice nucleation activity in snow algae (Chlamydomonas cf. nivalis and Chloromonas nivalis), lichen symbiotic algae (Trebouxia asymmetrica, Trebouxia erici and Trebouxia glomerata), and a mesophilic strain (Chlamydomonas reinhardti) were evaluated. Ice nucleation activity was measured using the freezing droplet method. Measurements were performed using suspensions of cells of A750 (absorbance at 750 nm) ~ 1, 0.1, 0.01 and 0.001 dilutions for each strain. The algae had lower ice nucleation activity, with the exception of Chloromonas nivalis contaminated by bacteria. The supercooling points of the snow algae were higher than those of lichen photobionts. The supercooling points of both, mesophilic and snow Chlamydomonas strains were similar. The lower freezing temperatures of the lichen algae may reflect either the more extreme and more variable environmental conditions of the original localities or the different cellular structure of the strains examined.

  6. Formosa haliotis sp. nov., a brown-alga-degrading bacterium isolated from the gut of the abalone Haliotis gigantea.

    PubMed

    Tanaka, Reiji; Cleenwerck, Ilse; Mizutani, Yukino; Iehata, Shunpei; Shibata, Toshiyuki; Miyake, Hideo; Mori, Tetsushi; Tamaru, Yutaka; Ueda, Mitsuyoshi; Bossier, Peter; Vandamme, Peter

    2015-12-01

    Four brown-alga-degrading, Gram-stain-negative, aerobic, non-flagellated, gliding and rod-shaped bacteria, designated LMG 28520T, LMG 28521, LMG 28522 and LMG 28523, were isolated from the gut of the abalone Haliotis gigantea obtained in Japan. The four isolates had identical random amplified polymorphic DNA patterns and grew optimally at 25 °C, at pH 6.0-9.0 and in the presence of 1.0-4.0 % (w/v) NaCl. Phylogenetic trees based on 16S rRNA gene sequences placed the isolates in the genus Formosa with Formosa algae and Formosa arctica as closest neighbours. LMG 28520T and LMG 28522 showed 100 % DNA-DNA relatedness to each other, 16-17 % towards F. algae LMG 28216T and 17-20 % towards F. arctica LMG 28318T; they could be differentiated phenotypically from these established species. The predominant fatty acids of isolates LMG 28520T and LMG 28522 were summed feature 3 (iso-C15 : 0 2-OH and/or C16 : 1ω7c), iso-C15 : 1 G and iso-C15 : 0. Isolate LMG 28520T contained menaquinone-6 (MK-6) as the major respiratory quinone and phosphatidylethanolamine, two unknown aminolipids and an unknown lipid as the major polar lipids. The DNA G+C content was 34.4 mol% for LMG 28520T and 35.5 mol% for LMG 28522. On the basis of their phylogenetic and genetic distinctiveness, and differential phenotypic properties, the four isolates are considered to represent a novel species of the genus Formosa, for which the name Formosa haliotis sp. nov. is proposed. The type strain is LMG 28520T ( = NBRC 111189T).

  7. Flavobacterium ahnfeltiae sp. nov., a new marine polysaccharide-degrading bacterium isolated from a Pacific red alga.

    PubMed

    Nedashkovskaya, Olga I; Balabanova, Larissa A; Zhukova, Natalia V; Kim, So-Jeong; Bakunina, Irina Y; Rhee, Sung-Keun

    2014-10-01

    A Gram-negative, aerobic, rod-shaped, motile by gliding and yellow-pigmented bacterium, designated strain 10Alg 130(T), that displayed the ability to destroy polysaccharides of red and brown algae, was isolated from the red alga Ahnfeltia tobuchiensis. The phylogenetic analysis based on 16S rRNA gene sequence placed the novel strain within the genus Flavobacterium, the type genus of the family Flavobacteriaceae, the phylum Bacteroidetes, with sequence similarities of 96.2 and 95.7 % to Flavobacterium jumunjiense KCTC 23618(T) and Flavobacterium ponti CCUG 58402(T), and 95.3-92.5 % to other recognized Flavobacterium species. The prevalent fatty acids of strain 10Alg 130(T) were iso-C15:0, iso-C15:0 3-OH, iso-C17:0 3-OH, C15:0 and iso-C17:1ω9c. The polar lipid profile consisted of phosphatidylethanolamine, two unknown aminolipids and three unknown lipids. The DNA G+C content of the type strain was 34.3 mol%. The new isolate and the type strains of recognized species of the genus Flavobacterium could strongly be distinguished by a number of phenotypic characteristics. A combination of the genotypic and phenotypic data showed that the algal isolate represents a novel species of the genus Flavobacterium, for which the name Flavobacterium ahnfeltiae sp. nov. is proposed. The type strain is 10Alg 130(T) (=KCTC 32467(T) = KMM 6686(T)).

  8. Host age and pathogen dosage impact cyst morphogenesis in the invertebrate pathogenic alga Helicosporidium sp. (Chlorophyta: Trebouxiophyceae).

    PubMed

    Denton, John S S; Lietze, Verena-Ulrike; Boucias, Drion G

    2009-09-01

    Helicosporidium sp. is a pathogenic alga that replicates in the hemolymph of various invertebrate hosts. Morphogenesis of the infectious life stage, the cyst, occurs in the infected host, but to date cannot be induced in vitro. Using larvae of the heterologous host Helicoverpa zea, we examined potential factors influencing pathogenicity and in vivo cyst production of the alga and the impact of infection on host survival. Factors tested were cyst dosage administered per os (ranging from 10(2) to 10(5) cysts per larva) and host age at exposure (third, fourth, and fifth larval instar). Cyst production occurred between 7 and 13days after treatment, regardless of host age at treatment. Increasing dosage increased both percent infection and mortality, but cyst production did not track the total infection response. Increasing host age at exposure mitigated dosage effects on infection and mortality and also elevated cyst production in later-treated larvae. Only the highest dosage produced a significant decrease in the overall time to death. Moderate cyst dosages and later host ages were most effective at regenerating Helicosporidium cysts.

  9. Purification and characterization of a novel alginate lyase from the marine bacterium Cobetia sp. NAP1 isolated from brown algae.

    PubMed

    Yagi, Hisashi; Fujise, Asako; Itabashi, Narumi; Ohshiro, Takashi

    2016-12-01

    The application of marine resources, instead of fossil fuels, for biomass production is important for building a sustainable society. Seaweed is valuable as a source of marine biomass for producing biofuels such as ethanol, and can be used in various fields. Alginate is an anionic polysaccharide that forms the main component of brown algae. Various alginate lyases (e.g. exo- and endo-types and oligoalginate lyase) are generally used to degrade alginate. We herein describe a novel alginate lyase, AlgC-PL7, which belongs to the polysaccharide lyase 7 family. AlgC-PL7 was isolated from the halophilic Gram-negative bacterium Cobetia sp. NAP1 collected from the brown algae Padina arborescens Holmes. The optimal temperature and pH for AlgC-PL7 activity were 45 °C and 8, respectively. Additionally, AlgC-PL7 was thermostable and salt-tolerant, exhibited broad substrate specificity, and degraded alginate into monosaccharides. Therefore, AlgC-PL7 is a promising enzyme for the production of biofuels.

  10. Epiphytic Terrestrial Algae (Trebouxia sp.) as a Biomarker Using the Free-Air-Carbon Dioxide-Enrichment (FACE) System.

    PubMed

    Ismail, Asmida; Marzuki, Sarah Diyana; Mohd Yusof, Nordiana Bakti; Buyong, Faeiza; Mohd Said, Mohd Nizam; Sigh, Harinder Rai; Zulkifli, Amyrul Rafiq

    2017-03-07

    The increasing concentration of CO₂ in the atmosphere has caused significant environmental changes, particularly to the lower plants such as terrestrial algae and lichens that alter species composition, and therefore can contribute to changes in community landscape. A study to understand how increased CO₂ in the atmosphere will affect algal density with minimal adjustment on its natural ecosystem, and the suitability of the algae to be considered as a biomarker, has been conducted. The current work was conducted in the Free-Air-Carbon Dioxide-Enrichment (FACE) system located in Universiti Kebangsaan Malaysia, Bangi, Malaysia. CO₂ was injected through special valves located along the ring surrounding specimen trees where 10 × 10 cm quadrats were placed. A total of 16 quadrats were randomly placed on the bark of 16 trees located inside the FACE system. This system will allow data collection on the effect of increased CO₂ without interfering or changing other parameters of the surrounding environment such as the wind speed, wind direction, humidity, and temperature. The initial density Trebouxia sp. was pre-determined on 1 March 2015, and the final density was taken slightly over a year later, on 15 March 2016. The exposure period of 380 days shed some light in understanding the effect of CO₂ on these non-complex, short life cycle lower plants. The results from this research work showed that the density of algae is significantly higher after 380 days exposure to the CO₂-enriched environment, at 408.5 ± 38.5 × 10⁴ cells/cm², compared to the control site at 176.5 ± 6.9 × 10⁴ cells/cm² (independent t-test, p < 0.001). The distance between the trees and the injector valves is negatively correlated. Quadrats located in the center of the circular ring recorded lower algal density compared to the ones closer to the CO₂ injector. Quadrat 16, which was nearing the end of the CO₂ valve injector, showed an exceptionally high algal density-2-fold higher

  11. Epiphytic Terrestrial Algae (Trebouxia sp.) as a Biomarker Using the Free-Air-Carbon Dioxide-Enrichment (FACE) System

    PubMed Central

    Ismail, Asmida; Marzuki, Sarah Diyana; Mohd Yusof, Nordiana Bakti; Buyong, Faeiza; Mohd Said, Mohd Nizam; Sigh, Harinder Rai; Zulkifli, Amyrul Rafiq

    2017-01-01

    The increasing concentration of CO2 in the atmosphere has caused significant environmental changes, particularly to the lower plants such as terrestrial algae and lichens that alter species composition, and therefore can contribute to changes in community landscape. A study to understand how increased CO2 in the atmosphere will affect algal density with minimal adjustment on its natural ecosystem, and the suitability of the algae to be considered as a biomarker, has been conducted. The current work was conducted in the Free-Air-Carbon Dioxide-Enrichment (FACE) system located in Universiti Kebangsaan Malaysia, Bangi, Malaysia. CO2 was injected through special valves located along the ring surrounding specimen trees where 10 × 10 cm quadrats were placed. A total of 16 quadrats were randomly placed on the bark of 16 trees located inside the FACE system. This system will allow data collection on the effect of increased CO2 without interfering or changing other parameters of the surrounding environment such as the wind speed, wind direction, humidity, and temperature. The initial density Trebouxia sp. was pre-determined on 1 March 2015, and the final density was taken slightly over a year later, on 15 March 2016. The exposure period of 380 days shed some light in understanding the effect of CO2 on these non-complex, short life cycle lower plants. The results from this research work showed that the density of algae is significantly higher after 380 days exposure to the CO2-enriched environment, at 408.5 ± 38.5 × 104 cells/cm2, compared to the control site at 176.5 ± 6.9 × 104 cells/cm2 (independent t-test, p < 0.001). The distance between the trees and the injector valves is negatively correlated. Quadrats located in the center of the circular ring recorded lower algal density compared to the ones closer to the CO2 injector. Quadrat 16, which was nearing the end of the CO2 valve injector, showed an exceptionally high algal density—2-fold higher than the average

  12. Bromophycoic acids: Bioactive natural products from a Fijian red alga Callophycus sp

    PubMed Central

    Teasdale, Margaret E.; Shearer, Tonya L.; Engel, Sebastian; Alexander, Troy S.; Fairchild, Craig R.; Prudhomme, Jacques; Torres, Manuel; Le Roch, Karine; Aalbersberg, William; Hay, Mark E.

    2012-01-01

    Bioassay-guided fractionation of extracts from a Fijian red alga in the genus Callophycus resulted in the isolation of five new compounds of the diterpene-benzoate class. Bromophycoic acids A-E (1–5) were characterized by NMR and mass spectroscopic analyses and represent two novel carbon skeletons, one with an unusual proposed biosynthesis. These compounds display a range of activities against human tumor cell lines, malarial parasite, and bacterial pathogens including low micromolar suppression of MRSA and VREF. PMID:22920243

  13. Distribution and occurrence of the insect pathogenic alga Helicosporidium sp. (Chlorophyta: Trebouxiophyceae) in the predator beetle Rhizophagus grandis G: yll. (Coleoptera: Rhizophagidae)-rearing laboratories.

    PubMed

    Yaman, M; Tosun, O; Aydın, C; Ertürk, O

    2011-01-01

    The distribution and occurrence of the insect pathogenic algae Helicosporidium sp. (Chlorophyta: Trebouxiophyceae) in the predator beetle Rhizophagus grandis (Coleoptera: Rhizophagidae)-rearing laboratories were studied and reported here for the first time. The insect pathogenic alga Helicosporidium sp. infection was observed in all R. grandis-rearing laboratories. The infection rate reached more than 20% which is significant among the samples in some R. grandis-rearing laboratories. The infection rates of the examined beetles showed noticeable differences between localities and years. There was no significant difference in the infection levels of male and female beetles. These results showed that Helicosporidium sp. is one of the factors that decrease efficiency of the R. grandis-rearing laboratories.

  14. Distinctive Architecture of the Chloroplast Genome in the Chlorodendrophycean Green Algae Scherffelia dubia and Tetraselmis sp. CCMP 881

    PubMed Central

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

    2016-01-01

    The Chlorodendrophyceae is a small class of green algae belonging to the core Chlorophyta, an assemblage that also comprises the Pedinophyceae, Trebouxiophyceae, Ulvophyceae and Chlorophyceae. Here we describe for the first time the chloroplast genomes of chlorodendrophycean algae (Scherffelia dubia, 137,161 bp; Tetraselmis sp. CCMP 881, 100,264 bp). Characterized by a very small single-copy (SSC) region devoid of any gene and an unusually large inverted repeat (IR), the quadripartite structures of the Scherffelia and Tetraselmis genomes are unique among all core chlorophytes examined thus far. The lack of genes in the SSC region is offset by the rich and atypical gene complement of the IR, which includes genes from the SSC and large single-copy regions of prasinophyte and streptophyte chloroplast genomes having retained an ancestral quadripartite structure. Remarkably, seven of the atypical IR-encoded genes have also been observed in the IRs of pedinophycean and trebouxiophycean chloroplast genomes, suggesting that they were already present in the IR of the common ancestor of all core chlorophytes. Considering that the relationships among the main lineages of the core Chlorophyta are still unresolved, we evaluated the impact of including the Chlorodendrophyceae in chloroplast phylogenomic analyses. The trees we inferred using data sets of 79 and 108 genes from 71 chlorophytes indicate that the Chlorodendrophyceae is a deep-diverging lineage of the core Chlorophyta, although the placement of this class relative to the Pedinophyceae remains ambiguous. Interestingly, some of our phylogenomic trees together with our comparative analysis of gene order data support the monophyly of the Trebouxiophyceae, thus offering further evidence that the previously observed affiliation between the Chlorellales and Pedinophyceae is the result of systematic errors in phylogenetic reconstruction. PMID:26849226

  15. Combined biocidal action of silver nanoparticles and ions against Chlorococcales (Scenedesmus quadricauda, Chlorella vulgaris) and filamentous algae (Klebsormidium sp.).

    PubMed

    Zouzelka, Radek; Cihakova, Pavlina; Rihova Ambrozova, Jana; Rathousky, Jiri

    2016-05-01

    Despite the extensive research, the mechanism of the antimicrobial and biocidal performance of silver nanoparticles has not been unequivocally elucidated yet. Our study was aimed at the investigation of the ability of silver nanoparticles to suppress the growth of three types of algae colonizing the wetted surfaces or submerged objects and the mechanism of their action. Silver nanoparticles exhibited a substantial toxicity towards Chlorococcales Scenedesmus quadricauda, Chlorella vulgaris, and filamentous algae Klebsormidium sp., which correlated with their particle size. The particles had very good stability against agglomeration even in the presence of multivalent cations. The concentration of silver ions in equilibrium with nanoparticles markedly depended on the particle size, achieving about 6 % and as low as about 0.1 % or even less for the particles 5 nm in size and for larger ones (40-70 nm), respectively. Even very limited proportion of small particles together with larger ones could substantially increase concentration of Ag ions in solution. The highest toxicity was found for the 5-nm-sized particles, being the smallest ones in this study. Their toxicity was even higher than that of silver ions at the same silver concentration. When compared as a function of the Ag(+) concentration in equilibrium with 5-nm particles, the toxicity of ions was at least 17 times higher than that obtained by dissolving silver nitrite (if not taking into account the effect of nanoparticles themselves). The mechanism of the toxicity of silver nanoparticles was found complex with an important role played by the adsorption of silver nanoparticles and the ions released from the particles on the cell surface. This mechanism could be described as some sort of synergy between nanoparticles and ions. While our study clearly showed the presence of this synergy, its detailed explanation is experimentally highly demanding, requiring a close cooperation between materials scientists

  16. Distinctive Architecture of the Chloroplast Genome in the Chlorodendrophycean Green Algae Scherffelia dubia and Tetraselmis sp. CCMP 881.

    PubMed

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

    2016-01-01

    The Chlorodendrophyceae is a small class of green algae belonging to the core Chlorophyta, an assemblage that also comprises the Pedinophyceae, Trebouxiophyceae, Ulvophyceae and Chlorophyceae. Here we describe for the first time the chloroplast genomes of chlorodendrophycean algae (Scherffelia dubia, 137,161 bp; Tetraselmis sp. CCMP 881, 100,264 bp). Characterized by a very small single-copy (SSC) region devoid of any gene and an unusually large inverted repeat (IR), the quadripartite structures of the Scherffelia and Tetraselmis genomes are unique among all core chlorophytes examined thus far. The lack of genes in the SSC region is offset by the rich and atypical gene complement of the IR, which includes genes from the SSC and large single-copy regions of prasinophyte and streptophyte chloroplast genomes having retained an ancestral quadripartite structure. Remarkably, seven of the atypical IR-encoded genes have also been observed in the IRs of pedinophycean and trebouxiophycean chloroplast genomes, suggesting that they were already present in the IR of the common ancestor of all core chlorophytes. Considering that the relationships among the main lineages of the core Chlorophyta are still unresolved, we evaluated the impact of including the Chlorodendrophyceae in chloroplast phylogenomic analyses. The trees we inferred using data sets of 79 and 108 genes from 71 chlorophytes indicate that the Chlorodendrophyceae is a deep-diverging lineage of the core Chlorophyta, although the placement of this class relative to the Pedinophyceae remains ambiguous. Interestingly, some of our phylogenomic trees together with our comparative analysis of gene order data support the monophyly of the Trebouxiophyceae, thus offering further evidence that the previously observed affiliation between the Chlorellales and Pedinophyceae is the result of systematic errors in phylogenetic reconstruction.

  17. Hemichloris antarctica, gen. et sp. nov. (Chlorococcales, Chlorophyta), a cryptoendolithic alga from Antarctica.

    PubMed

    Tschermak-Woess, E; Friedmann, E I

    1984-01-01

    Hemichloris antarctica gen. et sp. nov. (Oocystaceae, Chlorococcales) is characterized by a single, articulated, pyrenoid-less, thick saucer-shaped chloroplast, which generally fills less than half of the cell periphery. Multiplication is only by autospores. The species is psychrophilic and is damaged at temperatures above 20 degree C. Hemichloris antarctica is a member of the cryptoendolithic microbial community living in porous sandstone rocks of the Antarctica cold desert. It inhabits the zone below that of cryptoendolithic lichens and survives at extremely low light intensities. In the natural habitat, morphology is somewhat different from that in culture, as chloroplasts are smaller and without articulation, and the cells develop a gelatinous sheath.

  18. Wenyingzhuangia gracilariae sp. nov., a novel marine bacterium of the phylum Bacteroidetes isolated from the red alga Gracilaria vermiculophylla.

    PubMed

    Yoon, Jaewoo; Oku, Naoya; Kasai, Hiroaki

    2015-06-01

    A Gram-negative, strictly aerobic, beige-pigmented, non-motile, rod-shaped bacterial strain designated N5DB13-4(T) was isolated from the red alga Gracilaria vermiculophylla (Rhodophyta) collected at Sodegaura Beach, Chiba, Japan. Phylogenetic analyses based on the 16S rRNA gene sequence revealed that the novel isolate is affiliated with the family Flavobacteriaceae within the phylum Bacteroidetes and that it showed highest sequence similarity (97.3 %) to Wenyingzhuangia heitensis H-MN17(T). The hybridization values for DNA-DNA relatedness between the strains N5DB13-4(T) and W. heitensis H-MN17(T) were 34.1 ± 3.5 %, which is below the threshold accepted for the phylogenetic definition of a novel prokaryotic species. The DNA G+C content of strain N5DB13-4(T) was determined to be 31.8 mol%; MK-6 was identified as the major menaquinone; and the presence of iso-C15:0, iso-C15:0 3-OH and iso-C17:0 3-OH as the major (>10 %) cellular fatty acids. A complex polar lipid profile was present consisting of phosphatidylethanolamine, two unidentified glycolipids and four unidentified lipids. From the distinct phylogenetic position and combination of genotypic and phenotypic characteristics, the strain is considered to represent a novel species of the genus Wenyingzhuangia for which the name Wenyingzhuangia gracilariae sp. nov. is proposed. The type strain of W. gracilariae sp. nov. is N5DB13-4(T) (=KCTC 42246 (T)=NBRC 110602(T)).

  19. The Chlamydomonas genome project: a decade on

    PubMed Central

    Blaby, Ian K.; Blaby-Haas, Crysten; Tourasse, Nicolas; Hom, Erik F. Y.; Lopez, David; Aksoy, Munevver; Grossman, Arthur; Umen, James; Dutcher, Susan; Porter, Mary; King, Stephen; Witman, George; Stanke, Mario; Harris, Elizabeth H.; Goodstein, David; Grimwood, Jane; Schmutz, Jeremy; Vallon, Olivier; Merchant, Sabeeha S.; Prochnik, Simon

    2014-01-01

    The green alga Chlamydomonas reinhardtii is a popular unicellular organism for studying photosynthesis, cilia biogenesis and micronutrient homeostasis. Ten years since its genome project was initiated, an iterative process of improvements to the genome and gene predictions has propelled this organism to the forefront of the “omics” era. Housed at Phytozome, the Joint Genome Institute’s (JGI) plant genomics portal, the most up-to-date genomic data include a genome arranged on chromosomes and high-quality gene models with alternative splice forms supported by an abundance of RNA-Seq data. Here, we present the past, present and future of Chlamydomonas genomics. Specifically, we detail progress on genome assembly and gene model refinement, discuss resources for gene annotations, functional predictions and locus ID mapping between versions and, importantly, outline a standardized framework for naming genes. PMID:24950814

  20. Algimonas porphyrae gen. nov., sp. nov., a member of the family Hyphomonadaceae, isolated from the red alga Porphyra yezoensis.

    PubMed

    Fukui, Youhei; Abe, Mahiko; Kobayashi, Masahiro; Saito, Hiroaki; Oikawa, Hiroshi; Yano, Yutaka; Satomi, Masataka

    2013-01-01

    Three Gram-negative, stalked, motile bacteria, designated 0C-2-2(T), 0C-17 and LNM-3, were isolated from the red alga Porphyra yezoensis. 16S rRNA gene sequence analysis revealed that the three novel strains belonged to the family Hyphomonadaceae, and were closely related to Litorimonas taeanensis G5(T) (96.5 % 16S rRNA gene sequence similarity) and Hellea balneolensis 26III/A02/215(T) (94.3 %). The DNA G+C contents of the novel isolates (58.5-60.2 mol%) were clearly distinguished from those of L. taeanensis G5(T) (47.1 mol%) and H. balneolensis DSM 19091(T) (47.9 mol%). The G+C content of L. taeanensis G5(T) obtained in this study was quite different from a previous report (63.6 mol%). DNA-DNA hybridization experiments showed that the novel strains constituted a single species. Eleven phenotypic features of the three isolates differed from those of both related genera. The predominant respiratory quinone was ubiquinone-10 and the major fatty acid was C(18 : 1)ω7c. On the basis of this polyphasic taxonomic analysis, the novel strains represent a novel genus and species, for which the name Algimonas porphyrae gen. nov., sp. nov. is proposed. The type strain of Algimonas porphyrae is 0C-2-2(T) (= LMG 26424(T) = NBRC 108216(T)).

  1. Description of Persicirhabdus sediminis gen. nov., sp. nov., Roseibacillus ishigakijimensis gen. nov., sp. nov., Roseibacillus ponti sp. nov., Roseibacillus persicicus sp. nov., Luteolibacter pohnpeiensis gen. nov., sp. nov. and Luteolibacter algae sp. nov., six marine members of the phylum 'Verrucomicrobia', and emended descriptions of the class Verrucomicrobiae, the order Verrucomicrobiales and the family Verrucomicrobiaceae.

    PubMed

    Yoon, Jaewoo; Matsuo, Yoshihide; Adachi, Kyoko; Nozawa, Midori; Matsuda, Satoru; Kasai, Hiroaki; Yokota, Akira

    2008-04-01

    Ten pale-pink- and pale-yellow-pigmented, Gram-negative, non-motile, rod-shaped, chemoheterotrophic bacteria designated strains YM20-087T, YM21-151, MN1-741T, YM27-120T, YM26-010T, YM24-184, YM20-122, A4T-83T, A5J-41-2T and A5J-40 were isolated from various marine environments and were subjected to a polyphasic taxonomic investigation. Phylogenetic analyses based on 16S rRNA gene sequences indicated that these isolates belonged to the phylum 'Verrucomicrobia' (subdivision 1) and represented three independent lineages that were distinct from species of genera of the family Verrucomicrobiaceae with validly published names. The cell-wall peptidoglycan of these strains contained muramic acid and meso-diaminopimelic acid. Strains MN1-741T, YM27-120T, YM26-010T, YM24-184 and YM20-122 produced pinkish carotenoid pigments. On the basis of polyphasic taxonomic evidence, it was concluded that these strains should be classified within three new genera, Persicirhabdus gen. nov. (with one species, the type species Persicirhabdus sediminis sp. nov.), Roseibacillus gen. nov. (with three species; type species Roseibacillus ishigakijimensis sp. nov.) and Luteolibacter gen. nov. (with two species; type species Luteolibacter pohnpeiensis sp. nov.), of the family Verrucomicrobiaceae within the phylum 'Verrucomicrobia'. The names Persicirhabdus sediminis gen. nov., sp. nov. (type strain YM20-087T =MBIC08313T =KCTC 22039T), Roseibacillus ishigakijimensis gen. nov., sp. nov. (type strain MN1-741T =MBIC08315T =KCTC 12986T), Roseibacillus ponti sp. nov. (type strain YM27-120T =MBIC08316T =KCTC 12987T), Roseibacillus persicicus sp. nov. (type strain YM26-010T =MBIC08317T =KCTC 12988T), Luteolibacter pohnpeiensis gen. nov., sp. nov. (type strain A4T-83T =MBIC08322T =KCTC 22041T) and Luteolibacter algae sp. nov. (type strain A5J-41-2T =MBIC08320T =KCTC 22040T) are therefore proposed. Emended descriptions of the class Verrucomicrobiae, the order Verrucomicrobiales and the family

  2. Identification of an NADP/thioredoxin system in Chlamydomonas reinhardtii

    NASA Technical Reports Server (NTRS)

    Huppe, H. C.; Picaud, A.; Buchanan, B. B.; Miginiac-Maslow, M.

    1991-01-01

    The protein components of the NADP/thioredoxin system, NADP-thioredoxin reductase (NTR) and thioredoxin h, have been purified and characterized from the green alga, Chlamydomonas reinhardtii. The analysis of this system confirms that photoautotrophic Chlamydomonas cells resemble leaves in having both an NADP- and ferrodoxin-linked thioredoxin redox system. Chlamydomonas thioredoxin h, which is smaller on sodium dodecyl sulfate-polyacrylamide gel electrophoresis than thioredoxin m from the same source, cross-reacted with antisera to thioredoxin h from spinach (Spinacia oleracea L.) and wheat germ (Triticum vulgaris L.) but not with antisera to m or f thioredoxins. In these properties, the thioredoxin h resembled a thioredoxin from Chlamydomonas, designated Ch1, whose sequence was reported recently (P. Decottignies et al., 1991, Eur. J. Biochem. 198, 505-512). The differential reactivity of thioredoxin h with antisera was used to demonstrate that thioredoxin h is enriched outside the chloroplast. The NTR was purified from Chlamydomonas using thioredoxin h from the same source. Similar to its counterpart from other organisms, Chlamydomonas NTR had a subunit size of approx. 36 kDa and was specific for NADPH. Chlamydomonas NTR effectively reduced thioredoxin h from the same source but showed little activity with the other thioredoxins tested, including spinach thioredoxin h and Escherichia coli thioredoxin. Comparison of the reduction of Chlamydomonas thioredoxins m and h by each of the endogenous thioredoxin reductases, NTR and ferredoxin-thioredoxin reductase, revealed a differential specificity of each enzyme for thioredoxin. Thus, NTR showed increased activity with thioredoxin h and ferredoxin-thioredoxin reductase with thioredoxins m and f.

  3. Photomixing of chlamydomonas rheinhardtii suspensions

    NASA Astrophysics Data System (ADS)

    Dervaux, Julien; Capellazzi Resta, Marina; Abou, Bérengère; Brunet, Philippe

    2014-11-01

    Chlamydomonas rheinhardtii is a fast swimming unicellular alga able to bias its swimming direction in gradients of light intensity, an ability know as phototaxis. We have investigated experimentally both the swimming behavior of individual cells and the macroscopic response of shallow suspensions of these micro-organisms in response to a localized light source. At low light intensity, algae exhibit positive phototaxis and accumulate beneath the excitation light. In weakly concentrated thin layers, the balance between phototaxis and cell motility results in steady symmetrical patterns compatible with a purely diffusive model using effective diffusion coefficients extracted from the analysis of individual cell trajectories. However, at higher cell density and layer depth, collective effects induce convective flows around the light source. These flows disturb the cell concentration patterns which spread and may then becomes unstable. Using large passive tracer particles, we have characterized the velocity fields associated with this forced bioconvection and their dependence on the cell density and layer depth. By tuning the light distribution, this mechanism of photo-bioconvection allows a fine control over the local fluid flows, and thus the mixing efficiency, in algal suspensions.

  4. Identity and physiology of a new psychrophilic eukaryotic green alga, Chlorella sp., strain BI, isolated from a transitory pond near Bratina Island, Antarctica

    USGS Publications Warehouse

    Morgan-Kiss, R. M.; Ivanov, A.G.; Modla, S.; Czymmek, K.; Huner, N.P.A.; Priscu, J.C.; Lisle, J.T.; Hanson, T.E.

    2008-01-01

    Permanently low temperature environments are one of the most abundant microbial habitats on earth. As in most ecosystems, photosynthetic organisms drive primary production in low temperature food webs. Many of these phototrophic microorganisms are psychrophilic; however, functioning of the photosynthetic processes of these enigmatic psychrophiles (the 'photopsychrophiles') in cold environments is not well understood. Here we describe a new chlorophyte isolated from a low temperature pond, on the Ross Ice Shelf near Bratina Island, Antarctica. Phylogenetic and morphological analyses place this strain in the Chlorella clade, and we have named this new chlorophyte Chlorella BI. Chlorella BI is a psychrophilic species, exhibiting optimum temperature for growth at around 10??C. However, psychrophily in the Antarctic Chlorella was not linked to high levels of membrane-associated poly-unsaturated fatty acids. Unlike the model Antarctic lake alga, Chlamydomonas raudensis UWO241, Chlorella BI has retained the ability for dynamic short term adjustment of light energy distribution between photosystem II (PS II) and photosystem I (PS I). In addition, Chlorella BI can grow under a variety of trophic modes, including heterotrophic growth in the dark. Thus, this newly isolated photopsychrophile has retained a higher versatility in response to environmental change than other well studied cold-adapted chlorophytes. ?? 2008 Springer.

  5. Pretreatment for simultaneous production of total lipids and fermentable sugars from marine alga, Chlorella sp.

    PubMed

    Lee, Choon-Geun; Kang, Do-Hyung; Lee, Dong-Bog; Lee, Hyeon-Yong

    2013-11-01

    The goal of this study was to determine the optimal pretreatment process for the extraction of lipids and reducing sugars to facilitate the simultaneous production of biodiesel and bioethanol from the marine microalga Chorella sp. With a single pretreatment process, the optimal ultrasonication pretreatment process was 10 min at 47 KHz, and extraction yields of 6.5 and 7.1 (percentage, w/w) of the lipids and reducing sugars, respectively, were obtained. The optimal microwave pretreatment process was 10 min at 2,450 MHz, and extraction yields of 6.6 and 7.0 (percentage, w/w) of the lipids and reducing sugars, respectively, were obtained. Lastly, the optimal high-pressure homogenization pretreatment process was two cycles at a pressure of 20,000 psi, and extraction yields of 12.5 and 12.8 (percentage, w/w) of the lipids and reducing sugars, respectively, were obtained. However, because the single pretreatment processes did not markedly improve the extraction yields compared to the results of previous studies, a combination of two pretreatment processes was applied. The yields of lipids and reducing sugars from the combined application of the high-pressure homogenization process and the microwave process were 24.4 and 24.9 % (w/w), respectively, which was up to three times greater than the yields obtained using the single pretreatment processes. Furthermore, the oleic acid content, which is a fatty acid suitable for biodiesel production, was 23.39 % of the fatty acids (w/w). The contents of glucose and xylose, which are among the fermentable sugars useful for bioethanol production, were 77.5 and 13.3 % (w/w) of the fermentable sugars, respectively, suggesting the possibility of simultaneously producing biodiesel and bioethanol. Based on the results of this study, the combined application of the high-pressure homogenization and microwave pretreatment processes is the optimal method to increase the extraction yields of lipids and reducing sugars that are essential for

  6. [Carotenogenesis of five strains of the algae Dunaliella sp. (Chlorophyceae) isolated from Venezuelan hypersaline lagoons].

    PubMed

    Guevara, Miguel; Lodeiros, César; Gómez, Olga; Lemus, Nathalie; Núñez, Paulino; Romero, Lolymar; Vásquez, Aléikar; Rosales, Néstor

    2005-01-01

    We evaluated discontinuous cultures (Algal medium at 0.5 mM of NaNO3, and 27% NaCI) of five strains of Dunaliella sp. isolated from Venezuelan hypersaline lagoons (Araya, Coche, Peonia, Cumaraguas. and Boca Chica) and one strain from a reference collection (Dunaliella salina, LB1644). Cultures were maintained to 25+/-1 degrees C, with constant aeration, photoperiod 12:12, and two light intensities (195 and 390 microE.m(-2).s(-1)) during 30 days. Cell count was recorded on a daily basis using a Neubaüer camera. Totals of chlorophyll a and carotenoids were measured at the end of the experiment. The largest cellular densities were measured during the smallest light intensities. The strain with the largest cellular density was isolated from Boca Chica (8 xl0(6) and 2.5 xl0(6) cel.ml(-1) a 390 and 195microE.m(-2).s(-1), respectively). The increment of light intensity produced a significant reduction of growth rates in all strains. Totals of carotenoids by volume were as large as 390 microE.m(-2).s(-1). Strains LB 1644, from Coche and Araya were those that produced the largest amount of carotenoids (38.4; 32.8 and 21.0 microg.ml(-1), respectively). Differences total carotenoids by cell between treatments were significant. The largest concentration was 390 microE.m(-2).s(-1). The strains LB 1644 and Coche produced the highest values of carotenes (137.14 and 106.06 pg.cel(-1), respectively). Differences in the relation carotenoid:chlorophyll a between the strains at various light intensities was significant. Strains LB1644 presented the largest value of the relation carotenoids:chlorophyll a (20:1) at 195 microE.m(-2).s(-1). No significant differences were detected in the strain Coche (15:1). All the other strains showed relations lower than one. Our results suggest that the strains of Coche and Araya show potential to be used in the biotechnology of carotenoids production.

  7. Inhibition of tumor invasion and metastasis by calcium spirulan (Ca-SP), a novel sulfated polysaccharide derived from a blue-green alga, Spirulina platensis.

    PubMed

    Mishima, T; Murata, J; Toyoshima, M; Fujii, H; Nakajima, M; Hayashi, T; Kato, T; Saiki, I

    1998-08-01

    We have investigated the effect of calcium spirulan (Ca-SP) isolated from a blue-green alga, Spirulina platensis, which is a sulfated polysaccharide chelating calcium and mainly composed of rhamnose, on invasion of B16-BL6 melanoma, Colon 26 M3.1 carcinoma and HT-1080 fibrosarcoma cells through reconstituted basement membrane (Matrigel). Ca-SP significantly inhibited the invasion of these tumor cells through Matrigel/fibronectin-coated filters. Ca-SP also inhibited the haptotactic migration of tumor cells to laminin, but it had no effect on that to fibronectin. Ca-SP prevented the adhesion of B16-BL6 cells to Matrigel and laminin substrates but did not affect the adhesion to fibronectin. The pretreatment of tumor cells with Ca-SP inhibited the adhesion to laminin, while the pretreatment of laminin substrates did not. Ca-SP had no effect on the production and activation of type IV collagenase in gelatin zymography. In contrast, Ca-SP significantly inhibited degradation of heparan sulfate by purified heparanase. The experimental lung metastasis was significantly reduced by co-injection of B16-BL6 cells with Ca-SP. Seven intermittent i.v. injections of 100 microg of Ca-SP caused a marked decrease of lung tumor colonization of B16-BL6 cells in a spontaneous lung metastasis model. These results suggest that Ca-SP, a novel sulfated polysaccharide, could reduce the lung metastasis of B16-BL6 melanoma cells, by inhibiting the tumor invasion of basement membrane probably through the prevention of the adhesion and migration of tumor cells to laminin substrate and of the heparanase activity.

  8. Evaluation of an oil-producing green alga Chlorella sp. C2 for biological DeNOx of industrial flue gases.

    PubMed

    Zhang, Xin; Chen, Hui; Chen, Weixian; Qiao, Yaqin; He, Chenliu; Wang, Qiang

    2014-09-02

    NOx, a significant portion of fossil fuel flue gases, are among the most serious environmental issues in the world and must be removed in an additional costly gas treatment step. This study evaluated the growth of the green alga Chlorella sp. C2 under a nitrite-simulated NOx environment and the removal rates of actual flue gas fixed salts (FGFSs) from Sinopec's Shijiazhuang refinery along with lipid production. The results showed that nitrite levels lower than 176.5 mM had no significant adverse effects on the cell growth and photosynthesis of Chlorella sp. C2, demonstrating that this green alga could utilize nitrite and NOx as a nitrogen source. High concentrations of nitrite (88.25-176.5 mM) also resulted in the accumulation of neutral lipids. A 60% nitrite removal efficiency was obtained together with the production of 33% algae lipids when cultured with FGFS. Notably, the presence of nitrate in the FGFS medium significantly enhanced the nitrite removal capability, biomass and lipid production. Thus, this study may provide a new insight into the economically viable application of microalgae in the synergistic combination of biological DeNOx of industrial flue gases and biodiesel production.

  9. Cross-reconstitution of the extrinsic proteins and photosystem II complexes from Chlamydomonas reinhardtii and Spinacia oleracea.

    PubMed

    Suzuki, T; Ohta, H; Enami, I

    2005-06-01

    Cross-reconstitution of the extrinsic proteins and Photosystem II (PS II) from a green alga, Chlamydomonas reinhardtii, and a higher plant,Spinacia oleracea, was performed to clarify the differences of binding properties of the extrinsic proteins between these two species of organisms. (1) Chlamydomonas PsbP and PsbQ directly bound to Chlamydomonas PS II independent of the other extrinsic proteins but not to spinach PS II. (2) Chlamydomonas PsbP and PsbQ directly bound to the functional sites of Chlamydomonas PS II independent of the origins of PsbO, while spinach PsbP and PsbQ only bound to non-functional sites on Chlamydomonas PS II. (3) Both Chlamydomonas PsbP and spinach PsbP functionally bound to spinach PS II in the presence of spinach PsbO. (4) While Chlamydomonas PsbP functionally bound to spinach PS II in the presence of Chlamydomonas PsbO, spinach PsbP bound loosely to spinach PS II in the presence of Chlamydomonas PsbO with no concomitant restoration of oxygen evolution. (5) Chlamydomonas PsbQ bound to spinach PS II in the presence of Chlamydomonas PsbP and PsbO or spinach PsbO but not to spinach PS II in the presence of spinach PsbP and Chlamydomonas PsbO or spinach PsbO. (6) Spinach PsbQ did not bind to spinach PS II in the presence of Chlamydomonas PsbO and PsbP. On the basis of these results, we showed a simplified scheme for binding patterns of the green algal and higher plant extrinsic proteins with respective PS II.

  10. Toxic potential of iron oxide, CdS/Ag₂S composite, CdS and Ag₂S NPs on a fresh water alga Mougeotia sp.

    PubMed

    Jagadeesh, E; Khan, Behlol; Chandran, Preethy; Khan, S Sudheer

    2015-01-01

    Nanoparticles (NPs) are being used in many industries ranging from medical, textile, automobile, consumer products, etc. This may increase the probability of their (NPs) release into the environment and fresh water ecosystems. The present study focuses on testing the potential effect of iron oxide, nanocomposite of cadmium sulfide and silver sulfide, cadmium sulfide and silver sulfide nanoparticles (NPs) on a fresh water alga Mougeotia sp. as the model organism. The alga was treated with different concentrations of NPs (0.1-25 mg/L). The NPs exposure caused lipid peroxidation and ROS production, and suppressed the antioxidant defense system such as catalase, glutathione reductase, and superoxide dismutase. Adsorption of NPs on algal surface and membrane damage were confirmed through microscopic evaluation and increase in protein content in extracellular medium. The present investigation pointed out the ecological implications of NPs. The study warrants the need for regulatory agencies to monitor and regulate the use of NPs.

  11. Photoperiodic control of germination in the unicell Chlamydomonas

    NASA Astrophysics Data System (ADS)

    Suzuki, Lena; Johnson, Carl Hirschie

    2002-03-01

    Photoperiodic time measurement is a well-documented adaptation of multicellular plants and animals to seasonal changes in the environment, but it is unclear whether unicellular organisms can exhibit bona fide photoperiodic responses. We demonstrate that the occurrence of zygospore germination of the unicellular alga Chlamydomonas is a genuine photoperiodic response. Germination efficiency is enhanced in long days as compared with short days. While the total amount of light exposure influences the efficiency of germination, the photoperiod is a significant cue for germination. The developmental stage that senses the photoperiod is just prior to mating and during the first days of zygospore development, so there may be a critical window of zygospore maturation that is regulated by photoperiod. Because zygospores are resistant to freezing injury, whereas vegetative cells are not, it is likely that the suppression of germination by short days is an adaptive response for overwintering of Chlamydomonas. Therefore, Chlamydomonas is a single-celled organism that is capable of photoperiodic responses.

  12. Requirement of low oxidation-reduction potential for photosynthesis in a blue-green alga (Phormidium sp.).

    PubMed

    Weller, D; Doemel, W; Brock, T D

    1975-06-20

    Photosynthesis in a Phormidium species which forms dense conical-shaped structures in thermal springs is strongly inhibited by aeration but is stimulated by sulfide and other agents (cysteine, thioglycolate, sulfite) which lower the oxidation-reduction potential. The compact structures which this alga forms in nature may restrict oxygen penetration from the enviroment so that the anaerobic or microaerophilic conditions necessary ofr photosynthesis can develop. The alga may be defective in a regulatory mechanism that controls the reoxidation of reduced pyridine nucleotides formed during photosynthesis. It is suggested that other mat-forming and benthic blue-green algae may also prefer anaerobib conditions for growth and photosynthesis.

  13. Draft Genome Sequences of Achromobacter piechaudii GCS2, Agrobacterium sp. Strain SUL3, Microbacterium sp. Strain GCS4, Shinella sp. Strain GWS1, and Shinella sp. Strain SUS2 Isolated from Consortium with the Hydrocarbon-Producing Alga Botryococcus braunii.

    PubMed

    Jones, Katy J; Moore, Karen; Sambles, Christine; Love, John; Studholme, David J; Aves, Stephen J

    2016-01-14

    A variety of bacteria associate with the hydrocarbon-producing microalga Botryococcus braunii, some of which may influence its growth. We report here the genome sequences for Achromobacter piechaudii GCS2, Agrobacterium sp. strain SUL3, Microbacterium sp. strain GCS4, and Shinella sp. strains GWS1 and SUS2, isolated from a laboratory culture of B. braunii, race B, strain Guadeloupe.

  14. Characterization and optimization of hydrogen production by a salt water blue-green alga Oscillatoria sp. Miami BG 7. II - Use of immobilization for enhancement of hydrogen production

    NASA Technical Reports Server (NTRS)

    Phlips, E. J.; Mitsui, A.

    1986-01-01

    The technique of cellular immobilization was applied to the process of hydrogen photoproduction of nonheterocystous, filamentous marine blue-green alga, Oscillatoria sp. Miami BG 7. Immobilization with agar significantly improved the rate and longevity of hydrogen production, compared to free cell suspensions. Rates of H2 production in excess of 13 microliters H2 mg dry/wt h were observed and hydrogen production was sustained for three weeks. Immobilization also provided some stabilization to environmental variability and was adaptable to outdoor light conditions. In general, immobilization provides significant advantages for the production and maintenance of hydrogen photoproduction for this strain.

  15. Algibacter miyuki sp. nov., a member of the family Flavobacteriaceae isolated from leachate of a brown algae reservoir.

    PubMed

    Park, Sooyeon; Jung, Yong-Taek; Yoon, Jung-Hoon

    2013-08-01

    A Gram-negative, aerobic, non-flagellated, non-gliding and rod-shaped bacterial strain, designated WS-MY6(T), was isolated from a brown algae reservoir in South Korea. Strain WS-MY6(T) grew optimally at 25 °C, at pH 7.0-8.0 and in the presence of 2 % (w/v) NaCl. Neighbour-joining, maximum-likelihood and maximum-parsimony phylogenetic trees based on 16S rRNA gene sequences showed that strain WS-MY6(T) clustered with the type strains of Algibacter lectus and 'Algibacter undariae', showing 16S rRNA gene sequence similarity values of 98.1 and 98.4 %, respectively. It exhibited sequence similarities of 95.4-96.7 % to the type strains of the other Algibacter species, Pontirhabdus pectinovorans and Marinivirga aestuarii, whose reclassification into the genus Algibacter has been recently proposed. Strain WS-MY6(T) contained MK-6 as the predominant menaquinone and iso-C15:1 G, anteiso-C15:0 and iso-C17:0 3-OH as the major fatty acids. It contained phosphatidylethanolamine and two unidentified lipids as the major polar lipids. The DNA G + C content of strain WS-MY6(T) was 35.3 mol% and its DNA-DNA relatedness values with A. lectus KCTC 12103(T) and 'A. undariae' WS-MY9(T) was 21 and 13 %, respectively. The phylogenetic and genetic distinctiveness and differential phenotypic properties revealed that strain WS-MY6(T) is separate from existing Algibacter species. On the basis of the data presented, strain WS-MY6(T) is considered to represent a novel species of the genus Algibacter, for which the name Algibacter miyuki sp. nov. is proposed. The type strain is WS-MY6(T) (=KCTC 32382(T) =CECT 8300(T)).

  16. Algimonas ampicilliniresistens sp. nov., isolated from the red alga Porphyra yezoensis, and emended description of the genus Algimonas.

    PubMed

    Fukui, Youhei; Kobayashi, Masahiro; Saito, Hiroaki; Oikawa, Hiroshi; Yano, Yutaka; Satomi, Masataka

    2013-12-01

    Three strains (14A-2-7(T), 14A-3-1 and 14A-3) of Gram-stain-negative, prosthecate, motile bacteria were isolated from an algal medium supplemented with 10 mg ampicillin l(-1) (w/v), in which the red alga Porphyra yezoensis had been cultured. Based on the 16S rRNA gene sequence analysis, the three isolates formed a cluster with the genus Algimonas of the family Hyphomonadaceae. The sequences of the three isolates had high similarity with those of Algimonas porphyrae 0C-2-2(T) (97.6 % similarity) and Litorimonas taeanensis G5(T) (95.6 % similarity). The DNA G+C contents of the three isolates ranged from 54.3 to 55.0 mol%, which were more similar to that of A. porphyrae 0C-2-2(T) (58.5 mol%) than to that of L. taeanensis G5(T) (47.1 mol%). The DNA-DNA relatedness showed that the three isolates were representatives of the same species (88.1-94.0 % relatedness) and that strain 14A-2-7(T) was a representative of a different species from A. porphyrae 0C-2-2(T) and L. taeanensis G5(T) (1.2-8.6 % relatedness). The phenotypic characteristics of strain 14A-2-7(T) differed by 20 results and 30 results from A. porphyrae 0C-2-2(T) and L. taeanensis G5(T), respectively. The three isolates contained ubiquinone-10 as the predominant quinone and C18 : 1ω7c as the major fatty acid. Based on the polyphasic taxonomic analysis, the three isolates represent a novel species of the genus Algimonas, for which the name Algimonas ampicilliniresistens sp. nov. is proposed. The type strain is 14A-2-7(T) ( = LMG 26421(T) = NBRC 108219(T)). An emended description of the genus Algimonas is also proposed.

  17. Isolation of Chlamydomonas Flagella

    PubMed Central

    Craige, Branch; Brown, Jason M.; Witman, George B.

    2014-01-01

    A simple, scalable, and fast procedure for the isolation of Chlamydomonas flagella is described. Chlamydomonas can be synchronously deflagellated by treatment with chemicals, pH shock, or mechanical shear. The Basic Protocol describes the procedure for flagellar isolation using dibucaine to induce flagellar abscission; we also describe the pH shock method as an Alternate Protocol when flagellar regeneration is desirable. Sub-fractionation of the isolated flagella into axonemes and the membrane + matrix fraction is described in a Support Protocol. PMID:23728744

  18. Draft Genome Sequences of Achromobacter piechaudii GCS2, Agrobacterium sp. Strain SUL3, Microbacterium sp. Strain GCS4, Shinella sp. Strain GWS1, and Shinella sp. Strain SUS2 Isolated from Consortium with the Hydrocarbon-Producing Alga Botryococcus braunii

    PubMed Central

    Jones, Katy J.; Moore, Karen; Love, John

    2016-01-01

    A variety of bacteria associate with the hydrocarbon-producing microalga Botryococcus braunii, some of which may influence its growth. We report here the genome sequences for Achromobacter piechaudii GCS2, Agrobacterium sp. strain SUL3, Microbacterium sp. strain GCS4, and Shinella sp. strains GWS1 and SUS2, isolated from a laboratory culture of B. braunii, race B, strain Guadeloupe. PMID:26769927

  19. Effect of phosphorus fluctuation caused by river water dilution in eutrophic lake on competition between blue-green alga Microcystis aeruginosa and diatom Cyclotella sp.

    PubMed

    Amano, Yoshimasa; Sakai, Yusuke; Sekiya, Takumi; Takeya, Kimitaka; Taki, Kazuo; Machida, Motoi

    2010-01-01

    Tega-numa (Lake Tega) is one of the eutrophic lakes in Japan. For the improvement of water quality in Lake Tega, the North-chiba Water Conveyance Channel was constructed in 2000, which transfer water from Tone River into the lake. After 2000, the dominant species of diatoms, mainly Cyclotella sp., have been replacing blue-green algae, mainly Microcystis aeruginosa in Lake Tega. This transition of dominant species would be due to the dilution, but the detail mechanism has not been understood yet. This study examined the relationship between phosphorus fluctuation caused by river water dilution to Lake Tega and dominance of algal species, M. aeruginosa or Cyclotella sp. based on the single-species and the mixed-species culture experiments. The single-species culture experiment showed that the half-saturation constant and uptake rate of phosphorus were one order lower and seven times higher for M. aeruginosa than those for Cyclotella sp. These findings implied that M. aeruginosa would possess a potential for the growth and survival over Cyclotella sp. in the phosphorus limited condition. The superiority of M. aeruginosa was reflected in the outcome of the mixed-species culture experiment, i.e., dominance of M. aeruginosa, even phosphorus concentration was lowered to 0.01 mg-P/L. Therefore, it could be concluded that the decrease in phosphorus concentration due to the river water dilution to Lake Tega would be interpreted as a minor factor for the transition of dominant species from M. aeruginosa to Cyclotella sp.

  20. Acidophilic Green Alga Pseudochlorella sp. YKT1 Accumulates High Amount of Lipid Droplets under a Nitrogen-Depleted Condition at a Low-pH

    PubMed Central

    Hirooka, Shunsuke; Higuchi, Sumio; Uzuka, Akihiro; Nozaki, Hisayoshi; Miyagishima, Shin-ya

    2014-01-01

    Microalgal storage lipids are considered to be a promising source for next-generation biofuel feedstock. However, microalgal biodiesel is not yet economically feasible due to the high cost of production. One of the reasons for this is that the use of a low-cost open pond system is currently limited because of the unavoidable contamination with undesirable organisms. Extremophiles have an advantage in culturing in an open pond system because they grow in extreme environments toxic to other organisms. In this study, we isolated the acidophilic green alga Pseudochlorella sp. YKT1 from sulfuric acid mine drainage in Nagano Prefecture, Japan. The vegetative cells of YKT1 display the morphological characteristics of Trebouxiophyceae and molecular phylogenetic analyses indicated it to be most closely related to Pseudochlorella pringsheimii. The optimal pH and temperature for the growth of YKT1 are pH 3.0–5.0 and a temperature 20–25°C, respectively. Further, YKT1 is able to grow at pH 2.0 and at 32°C, which corresponds to the usual water temperature in the outdoors in summer in many countries. YKT1 accumulates a large amount of storage lipids (∼30% of dry weigh) under a nitrogen-depleted condition at low-pH (pH 3.0). These results show that acidophilic green algae will be useful for industrial applications by acidic open culture systems. PMID:25221913

  1. Crouania pumila sp. nov. (Callithamniaceae: Rhodophyta), a new species of marine red algae from the Seaflower International Biosphere Reserve, Caribbean Colombia.

    PubMed

    Gavio, Brigitte; Reyes-Gómez, Viviana P; Wynne, Michael J

    2013-09-01

    In the Colombian Caribbean, the marine macroalgal flora of the Seaflower International Biosphere Reserve has been little studied, despite its ecological importance. Historical records have reported only 201 macroalgae species within its area of almost 350,000 km2. However, recent surveys have shown a diversity of small algae previously overlooked. With the aim to determine the macroalgal diversity in the Reserve, we undertook field surveys in different ecosystems: coral reefs, seagrass beds, and rocky and sandy substrates, at different depths, from intertidal to 37 m. During these field surveys, we collected a small described species belonging to the genus Crouania (Callithamniaceae, Rhodophyta), Crouania pumila sp. nov. that is decribed in this paper. This new species was distinguished from other species of the genus by a distinctive suite of traits including its diminutive size (to only 3.5 mm in length), its decumbent, slightly calcified habit (epiphytic on other algae), its ramisympodial branching, the ecorticate main axes, and the elongate shape of the terminal cells of the cortical filaments. The observations were provided for both female (cystocarpic) and tetrasporangiate thalli; however, male thalli were not seen. Further studies have to be undertaken in this Reserve in order to carry out other macroalgal analysis and descriptions.

  2. Absence of lutein, violaxanthin and neoxanthin affects the functional chlorophyll antenna size of photosystem-II but not that of photosystem-I in the green alga Chlamydomonas reinhardtii.

    PubMed

    Polle, J E; Niyogi, K K; Melis, A

    2001-05-01

    Chlamydomonas reinhardtii double mutant npq2 lor1 lacks the beta, epsilon-carotenoids lutein and loroxanthin as well as all beta,beta-epoxycarotenoids derived from zeaxanthin (e.g. violaxanthin and neoxanthin). Thus, the only carotenoids present in the thylakoid membranes of the npq2 lor1 cells are beta-carotene and zeaxanthin. The effect of these mutations on the photochemical apparatus assembly and function was investigated. In cells of the mutant strain, the content of photosystem-II (PSII) and photosystem-I (PSI) was similar to that of the wild type, but npq2 lor1 had a significantly smaller PSII light-harvesting Chl antenna size. In contrast, the Chl antenna size of PSI was not truncated in the mutant. SDS-PAGE and Western blot analysis qualitatively revealed the presence of all LHCII and LHCI apoproteins in the thylakoid membrane of the mutant. The results showed that some of the LHCII and most of the LHCI were assembled and functionally connected with PSII and PSI, respectively. Photon conversion efficiency measurements, based on the initial slope of the light-saturation curve of photosynthesis and on the yield of Chl a fluorescence in vivo, showed similar efficiencies. However, a significantly greater light intensity was required for the saturation of photosynthesis in the mutant than in the wild type. It is concluded that zeaxanthin can successfully replace lutein and violaxanthin in most of the functional light-harvesting antenna of the npq2 lor1 mutant.

  3. Most microRNAs in the single-cell alga Chlamydomonas reinhardtii are produced by Dicer-like 3-mediated cleavage of introns and untranslated regions of coding RNAs

    PubMed Central

    Valli, Adrian A.; Santos, Bruno A.C.M.; Hnatova, Silvia; Bassett, Andrew R.; Molnar, Attila; Chung, Betty Y.; Baulcombe, David C.

    2016-01-01

    We describe here a forward genetic screen to investigate the biogenesis, mode of action, and biological function of miRNA-mediated RNA silencing in the model algal species, Chlamydomonas reinhardtii. Among the mutants from this screen, there were three at Dicer-like 3 that failed to produce both miRNAs and siRNAs and others affecting diverse post-biogenesis stages of miRNA-mediated silencing. The DCL3-dependent siRNAs fell into several classes including transposon- and repeat-derived siRNAs as in higher plants. The DCL3-dependent miRNAs differ from those of higher plants, however, in that many of them are derived from mRNAs or from the introns of pre-mRNAs. Transcriptome analysis of the wild-type and dcl3 mutant strains revealed a further difference from higher plants in that the sRNAs are rarely negative switches of mRNA accumulation. The few transcripts that were more abundant in dcl3 mutant strains than in wild-type cells were not due to sRNA-targeted RNA degradation but to direct DCL3 cleavage of miRNA and siRNA precursor structures embedded in the untranslated (and translated) regions of the mRNAs. Our analysis reveals that the miRNA-mediated RNA silencing in C. reinhardtii differs from that of higher plants and informs about the evolution and function of this pathway in eukaryotes. PMID:26968199

  4. The effects of irradiance levels and spectral composition on mating strategies in the snow alga, Chloromonas sp.-D, from the Tughill Plateau, New York State

    NASA Astrophysics Data System (ADS)

    Hoham, Ronald W.; Schlag, Erin M.; Kang, Jennifer Y.; Hasselwander, Andrew J.; Behrstock, Alissa F.; Blackburn, Ian R.; Johnson, Rurik C.; Roemer, Stephen C.

    1998-07-01

    Studies have related changes in snow albedo to snow crystal structure and to the presence of surface debris (i.e. pine needles), but there has been less attention given to the existence of algae in snow. An increase in the number of snow algae could also decrease albedo and increase snowmelt rates. The primary purpose of this paper is to document how solar irradiance serves to control the developing stages of algae in snow. Snow algae do not appear near the surface until there is meltwater in the snowpack. Low levels of solar irradiance penetrate through the snowpack and germinate snow algal resting stages that lie underneath, and snow algal growth is enhanced by available gases and nutrients. Flagellate cells swim through the snowpack in the meltwater around the snow crystals, and cells are positioned according to irradiance and spectral differences. In this study, Chloromonas sp.-D strains 582C and 582D, isolated from the upper 20 cm of snowpacks in the Tughill Plateau, Whetstone Gulf State Park, NY, were used to investigate mating strategies under different irradiance levels and spectral compositions in the laboratory, and the irradiance levels used in the experiments correlated with those recorded from the upper 20 cm of snow. Using similar irradiance levels, blue light regimes produced more matings than green and red light regimes. There were no trends in mating when comparing green and red light regimes. When red light regimes of higher photon irradiance (85 mol m-2 s-1) were compared with those of blue light regimes of lower irradiance (30 mol m-2 s-1), more mating occurred under red light. A photon irradiance of 95 mol m-2 s-1 [photosynthetically active radiation (PAR) of 400-700 nm] produced the most mating under both wide-spectrum (WS) and cool-white (CW) regimes, but more mating occurred under CW in all irradiances tested. Mating pairs of three types were observed: oblong-oblong (o-o), oblong-sphere (o-s) and sphere-sphere (s-s). Cell packs that produced

  5. Inhibitory effects of soluble algae products (SAP) released by Scenedesmus sp. LX1 on its growth and lipid production.

    PubMed

    Zhang, Tian-Yuan; Yu, Yin; Wu, Yin-Hu; Hu, Hong-Ying

    2013-10-01

    Soluble algal products (SAP) accumulated in culture medium via water reuse may affect the growth of microalga during the cultivation. Scenedesmus sp. LX1, a freshwater microalga, was used in this study to investigate the effect of SAP on growth and lipid production of microalga. Under the SAP concentrations of 6.4-25.8 mg L(-1), maximum algal density (K) and maximum growth rate (Rmax) of Scenedesmus sp. LX1 were decreased by 50-80% and 35-70% compared with the control group, respectively. The effect of SAP on lipid accumulation of Scenedesmus sp. LX1 was non-significant. According to hydrophilic-hydrophobic and acid-base properties, SAP was fractionized into six fractions. All of the fractions could inhibit the growth of Scenedesmus sp. LX1. Organic bases (HIB, HOB) and hydrophilic acids (HIA) showed the strongest inhibition. HIA could also decrease the lipid content of Scenedesmus sp. LX1 by 59.2%. As the inhibitory effect, SAP should be seriously treated before water reuse.

  6. Use of biofuel by-product from the green algae Desmochloris sp. and diatom Nanofrustulum sp. meal in diets for nile tilapia Oreochromis niloticus

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Algal by-product meals from the Hawaiian biofuels industry were evaluated as protein ingredients in diets for juveniles of Nile tilapia (Oreochromis niloticus). Four experimental diets were formulated to contain 40% protein and were made with fish meal, soybean meal, whole diatom (Nanofrustulum sp.)...

  7. Cryopreservation of economically valuable marine micro-algae in the classes Bacillariophyceae, Chlorophyceae, Cyanophyceae, Dinophyceae, Haptophyceae, Prasinophyceae, and Rhodophyceae.

    PubMed

    Rhodes, Lesley; Smith, John; Tervit, Robin; Roberts, Rodney; Adamson, Janet; Adams, Serean; Decker, Marie

    2006-02-01

    The ability to routinely cryopreserve micro-algal species reduces costs associated with maintaining large culture collections and reduces the risks of losing particular strains or species through contamination and genetic drift. Cryopreservation is also a useful adjunct in aquaculture hatcheries for strains of micro-algae where the nutritional status may change as a result of continuous sub-culture. In this study, cryopreservation of isolates from seven micro-algal classes was investigated. Successful candidates included the marine dinoflagellates Amphidinium carterae, Amphidinium trulla, and Gymnodinium simplex, and the haptophytes Chrysochromulina simplex, Prymnesium parvum, Prymnesium parvum f. patelliferum, Isochrysis galbana, and Pavlova lutheri. Also successfully cryopreserved were the planktonic diatoms Chaetoceros calcitrans, Chaetoceros muelleri, Chaetoceros sp., and the benthic Nitzschia ovalis, the chlorophyte Chlamydomonas coccoides, the rhodophyte Porphyridium purpureum, the prasinophytes Tetraselmis chuii, and Tetraselmis suecica, and the cyanophytes Raphidiopsis sp., and Aphanizomenon flos-aquae. All species were successfully cryopreserved using 15% Me2SO.

  8. Azotobacter vinelandii siderophore can provide nitrogen to support the culture of the green algae Neochloris oleoabundans and Scenedesmus sp. BA032.

    PubMed

    Villa, Juan A; Ray, Erin E; Barney, Brett M

    2014-02-01

    Microalgae are viewed as a potential future agricultural and biofuel feedstock and also provide an ideal biological means of carbon sequestration based on rapid growth rates and high biomass yields. Any potential improvement using high-yield microalgae to fix carbon will require additional fertilizer inputs to provide the necessary nitrogen required for protein and nucleotide biosynthesis. The free-living diazotroph Azotobacter vinelandii can fix nitrogen under aerobic conditions in the presence of reduced carbon sources such as sucrose or glycerol and is also known to produce a variety of siderophores to scavenge different metals from the environment. In this study, we identified two strains of green algae, Neochloris oleoabundans and Scenedesmus sp. BA032, that are able to utilize the A. vinelandii siderophore azotobactin as a source of nitrogen to support growth. When grown in a co-culture, S. sp. BA032 and N. oleoabundans obtained the nitrogen required for growth through the association with A. vinelandii. These results, indicating a commensalistic relationship, provide a proof of concept for developing a mutualistic or symbiotic relationship between these two species using siderophores as a nitrogen shuttle and might further indicate an additional fate of siderophores in the environment.

  9. Proton gradient regulation 5-mediated cyclic electron flow under ATP- or redox-limited conditions: a study of ΔATpase pgr5 and ΔrbcL pgr5 mutants in the green alga Chlamydomonas reinhardtii.

    PubMed

    Johnson, Xenie; Steinbeck, Janina; Dent, Rachel M; Takahashi, Hiroko; Richaud, Pierre; Ozawa, Shin-Ichiro; Houille-Vernes, Laura; Petroutsos, Dimitris; Rappaport, Fabrice; Grossman, Arthur R; Niyogi, Krishna K; Hippler, Michael; Alric, Jean

    2014-05-01

    The Chlamydomonas reinhardtii proton gradient regulation5 (Crpgr5) mutant shows phenotypic and functional traits similar to mutants in the Arabidopsis (Arabidopsis thaliana) ortholog, Atpgr5, providing strong evidence for conservation of PGR5-mediated cyclic electron flow (CEF). Comparing the Crpgr5 mutant with the wild type, we discriminate two pathways for CEF and determine their maximum electron flow rates. The PGR5/proton gradient regulation-like1 (PGRL1) ferredoxin (Fd) pathway, involved in recycling excess reductant to increase ATP synthesis, may be controlled by extreme photosystem I acceptor side limitation or ATP depletion. Here, we show that PGR5/PGRL1-Fd CEF functions in accordance with an ATP/redox control model. In the absence of Rubisco and PGR5, a sustained electron flow is maintained with molecular oxygen instead of carbon dioxide serving as the terminal electron acceptor. When photosynthetic control is decreased, compensatory alternative pathways can take the full load of linear electron flow. In the case of the ATP synthase pgr5 double mutant, a decrease in photosensitivity is observed compared with the single ATPase-less mutant that we assign to a decreased proton motive force. Altogether, our results suggest that PGR5/PGRL1-Fd CEF is most required under conditions when Fd becomes overreduced and photosystem I is subjected to photoinhibition. CEF is not a valve; it only recycles electrons, but in doing so, it generates a proton motive force that controls the rate of photosynthesis. The conditions where the PGR5 pathway is most required may vary in photosynthetic organisms like C. reinhardtii from anoxia to high light to limitations imposed at the level of carbon dioxide fixation.

  10. The High Efficiency of Photosystem I in the Green Alga Chlamydomonas reinhardtii Is Maintained after the Antenna Size Is Substantially Increased by the Association of Light-harvesting Complexes II*

    PubMed Central

    Le Quiniou, Clotilde; van Oort, Bart; Drop, Bartlomiej; van Stokkum, Ivo H. M.; Croce, Roberta

    2015-01-01

    Photosystems (PS) I and II activities depend on their light-harvesting capacity and trapping efficiency, which vary in different environmental conditions. For optimal functioning, these activities need to be balanced. This is achieved by redistribution of excitation energy between the two photosystems via the association and disassociation of light-harvesting complexes (LHC) II, in a process known as state transitions. Here we study the effect of LHCII binding to PSI on its absorption properties and trapping efficiency by comparing time-resolved fluorescence kinetics of PSI-LHCI and PSI-LHCI-LHCII complexes of Chlamydomonas reinhardtii. PSI-LHCI-LHCII of C. reinhardtii is the largest PSI supercomplex isolated so far and contains seven Lhcbs, in addition to the PSI core and the nine Lhcas that compose PSI-LHCI, together binding ∼320 chlorophylls. The average decay time for PSI-LHCI-LHCII is ∼65 ps upon 400 nm excitation (15 ps slower than PSI-LHCI) and ∼78 ps upon 475 nm excitation (27 ps slower). The transfer of excitation energy from LHCII to PSI-LHCI occurs in ∼60 ps. This relatively slow transfer, as compared with that from LHCI to the PSI core, suggests loose connectivity between LHCII and PSI-LHCI. Despite the relatively slow transfer, the overall decay time of PSI-LHCI-LHCII remains fast enough to assure a 96% trapping efficiency, which is only 1.4% lower than that of PSI-LHCI, concomitant with an increase of the absorption cross section of 47%. This indicates that, at variance with PSII, the design of PSI allows for a large increase of its light-harvesting capacities. PMID:26504081

  11. The High Efficiency of Photosystem I in the Green Alga Chlamydomonas reinhardtii Is Maintained after the Antenna Size Is Substantially Increased by the Association of Light-harvesting Complexes II.

    PubMed

    Le Quiniou, Clotilde; van Oort, Bart; Drop, Bartlomiej; van Stokkum, Ivo H M; Croce, Roberta

    2015-12-18

    Photosystems (PS) I and II activities depend on their light-harvesting capacity and trapping efficiency, which vary in different environmental conditions. For optimal functioning, these activities need to be balanced. This is achieved by redistribution of excitation energy between the two photosystems via the association and disassociation of light-harvesting complexes (LHC) II, in a process known as state transitions. Here we study the effect of LHCII binding to PSI on its absorption properties and trapping efficiency by comparing time-resolved fluorescence kinetics of PSI-LHCI and PSI-LHCI-LHCII complexes of Chlamydomonas reinhardtii. PSI-LHCI-LHCII of C. reinhardtii is the largest PSI supercomplex isolated so far and contains seven Lhcbs, in addition to the PSI core and the nine Lhcas that compose PSI-LHCI, together binding ∼ 320 chlorophylls. The average decay time for PSI-LHCI-LHCII is ∼ 65 ps upon 400 nm excitation (15 ps slower than PSI-LHCI) and ∼ 78 ps upon 475 nm excitation (27 ps slower). The transfer of excitation energy from LHCII to PSI-LHCI occurs in ∼ 60 ps. This relatively slow transfer, as compared with that from LHCI to the PSI core, suggests loose connectivity between LHCII and PSI-LHCI. Despite the relatively slow transfer, the overall decay time of PSI-LHCI-LHCII remains fast enough to assure a 96% trapping efficiency, which is only 1.4% lower than that of PSI-LHCI, concomitant with an increase of the absorption cross section of 47%. This indicates that, at variance with PSII, the design of PSI allows for a large increase of its light-harvesting capacities.

  12. Metabolism of D-lactate and structurally related organic acids in Chlamydomonas reinhardtii

    SciTech Connect

    Husic, D.W.

    1986-01-01

    During the initial minutes of anaerobiosis, /sup 14/C-labeled D-lactate, derived from the photosynthetic sugar phosphate pool, accumulated in the unicellular green alga, Chlamydomonas reinhardtii. The production of the D-isomer of lactate by algae is in contrast to plant and mammalian cells in which L-lactate is formed. After initial lactate formation, Chlamydomonas exhibits a mixed-acid type fermentation, thereby avoiding lactate accumulation and enabling the cells to tolerate extended periods of anaerobiosis. A pyruvate reductase which catalyzes the formation of D-lactate in Chlamydomonas was partially purified and characterized. Lactate produced anaerobically was metabolized only when Chlamydomonas cells were returned to aerobic conditions, and reoxidation of the D-lactate was apparently catalyzed by a mitochondrial membrane-bound dehydrogenase, rather than by the soluble pyruvate reductase. Mutants of Chlamydomonas, deficient in mitochondrial respiration, were used to demonstrate that lactate metabolism was linked to the mitochondrial electron transport chain. In addition, the oxidation of glycolate, a structural analog of lactate, was also linked to mitochondrial electron transport in vivo.

  13. Strategies to facilitate transgene expression in Chlamydomonas reinhardtii.

    PubMed

    Eichler-Stahlberg, Alke; Weisheit, Wolfram; Ruecker, Ovidiu; Heitzer, Markus

    2009-03-01

    The unicellular green alga Chlamydomonas reinhardtii has been identified as a promising organism for the production of recombinant proteins. While during the last years important improvements have been developed for the production of proteins within the chloroplast, the expression levels of transgenes from the nuclear genome were too low to be of biotechnological importance. In this study, we integrated endogenous intronic sequences into the expression cassette to enhance the expression of transgenes in the nucleus. The insertion of one or more copies of intron sequences from the Chlamydomonas RBCS2 gene resulted in increased expression levels of a Renilla-luciferase gene used as a reporter. Although any of the three RBCS2 introns alone had a positive effect on expression, their integration in their physiological number and order created an over-proportional stimulating effect observed in all transformants. The secretion of the luciferase protein into the medium was achieved by using the export sequence of the Chlamydomonas ARS2 gene in a cell wall deficient strain and Renilla-luciferase could be successfully concentrated with the help of attached C-terminal protein tags. Similarly, a codon adapted gene variant for human erythropoietin (crEpo) was expressed as a protein of commercial relevance. Extracellular erythropoietin produced in Chlamydomonas showed a molecular mass of 33 kDa probably resulting from post-translational modifications. Both, the increased expression levels of transgenes by integration of introns and the isolation of recombinant proteins from the culture medium are important steps towards an extended biotechnological use of this alga.

  14. Cytoduction in Chlamydomonas reinhardtii.

    PubMed Central

    Matagne, R F; Remacle, C; Dinant, M

    1991-01-01

    After conjugation between Chlamydomonas gametes of opposite mating type, a transient dikaryon is formed. The two nuclei fuse within 4-6 hr after mating. The young diploid zygote differentiates into dormant zygospore competent to complete meiosis, or more rarely (2-10% of cases) it undergoes mitosis to produce a stable diploid progeny. We here bring genetical, biochemical, and cytological evidence that among the mitotic zygotes, a large proportion of them undergo cytokinesis without fusion of the nuclei-a process that has been termed "cytoduction." By using appropriate genetic markers, haploid cytoductants that possess the nuclear genotype of one parent and the chloroplast marker of the other parent can easily be isolated. Genetical analysis and hybridization experiments moreover show that many haploid cytoductants transmit the chloroplast DNA molecules of both parents and that, as in diploids, these DNA copies occasionally recombine. This process of cytoduction extends the life cycle of Chlamydomonas and provides new tools for its genetic analysis. Images PMID:1871143

  15. Reparation in unicellular green algae during chronic exposure to the action of mutagenic factors. II. Restoration of single-stranded DNA breaks following exposure of Chlamydomonas reinchardii to gamma-irradiation

    SciTech Connect

    Sergeeva, S.A.; Ptitsina, S.N.; Shevchenko, V.A.

    1986-12-01

    The restoration of single-stranded breaks in the DNA in different strains of unicellular green algae (chlamydomonads) during chronic exposure to the action of mutagenic factors following ..gamma..-irradiation was investigated. It was shown that the restoration of DNA breaks was most effective in the case of strain M ..gamma../sup mt/sup +//, which is resistant to radiation. Strains, that were sensitive to UV irradiation showed a similar order of DNA break restoration as the wild-type strain. Strain UVS-1 showed a higher level of restoration than the wild-type strain. The data indicated that chlamydomonads have different pathways of reparation, which lead to the restoration of breaks induced by ..gamma..-irradiation and UV-rays.

  16. Analysis of cargo transport by IFT and GFP imaging of IFT in Chlamydomonas.

    PubMed

    Diener, Dennis

    2009-01-01

    Chlamydomonas reinhardtii is the organism in which intraflagellar transport (IFT) was first visualized and in which the composition of IFT particles was originally elucidated. As the universality of IFT among ciliated/flagellated cells was uncovered, the diversity of organisms used to study IFT has grown. Still, because of the ease of isolation of flagella from Chlamydomonas and the battery of temperature-sensitive mutants affecting IFT proteins and motors, this unicellular alga remains the principal model for biochemical studies of IFT motors and cargo; furthermore, the long, exposed flagella of this cell are ideally suited for observing IFT in real time with GFP-tagged components of IFT.

  17. Hydrogen evolution as a consumption mode of reducing equivalents in green algal fermentation. [Chlamydomonas reinhardii; Chlorella pyrenoidosa; Chlorococcum minutum

    SciTech Connect

    Ohta, S.; Miyamoto, K.; Miura, Y.

    1987-04-01

    Dark anaerobic fermentation in the green algae Chlamydomonas MGA 161, Chlamydomonas reinhardtii, Chlorella pyrenoidosa, and Chlorococcum minutum was studied. Their isolate, Chlamydomonas MGA 161, was unusual in having high H/sub 2/ but almost no formate. The fermentation pattern in Chlamydomonas MGA 161 was altered by changes in the NaCl or NH/sub 4/Cl concentration. Glycerol formation increased at low (0.1%) and high (7%) NaCl concentrations starch degradation, and formation of ethanol, H/sub 2/, and CO/sub 2/ increased with the addition of NH/sub 4/Cl to above 5 millimolar in N-deficient cells. C. reinhardtii and C.pyrenoidosa exhibited a very similar anaerobic metabolism, forming formate, acetate and ethanol in a ratio of about 2:2:1. C. minimum was also unusual in forming acetate, glycerol, and CO/sub 2/ as its main products, with H/sub 2/, formate, and ethanol being formed in negligible amounts. In the presence of CO, ethanol formation increased twofold in Chlamydomonas MGA 161 and C. reinhardtii, but the fermentation pattern in C. minimum did not change. An experiment with hypophosphite addition showed that dark H/sub 2/ evolution of the Escherichia coli type could be ruled out in Chlamydomonas MGA 161 and C. reinhardtii. Among the green algae investigated, three fermentation types were identified by the distribution pattern of the end products, which reflected the consumption model of reducing equivalents in the cells.

  18. Enhancement of Biodiesel Production from Marine Alga, Scenedesmus sp. through In Situ Transesterification Process Associated with Acidic Catalyst

    PubMed Central

    Kim, Ga Vin; Choi, WoonYong; Kang, DoHyung; Lee, ShinYoung; Lee, HyeonYong

    2014-01-01

    The aim of this study was to increase the yield of biodiesel produced by Scenedesmus sp. through in situ transesterification by optimizing various process parameters. Based on the orthogonal matrix analysis for the acidic catalyst, the effects of the factors decreased in the order of reaction temperature (47.5%) > solvent quantity (26.7%) > reaction time (17.5%) > catalyst amount (8.3%). Based on a Taguchi analysis, the effects of the factors decreased in the order of solvent ratio (34.36%) > catalyst (28.62%) > time (19.72%) > temperature (17.32%). The overall biodiesel production appeared to be better using NaOH as an alkaline catalyst rather than using H2SO4 in an acidic process, at 55.07 ± 2.18% (based on lipid weight) versus 48.41 ± 0.21%. However, in considering the purified biodiesel, it was found that the acidic catalyst was approximately 2.5 times more efficient than the alkaline catalyst under the following optimal conditions: temperature of 70°C (level 2), reaction time of 10 hrs (level 2), catalyst amount of 5% (level 3), and biomass to solvent ratio of 1 : 15 (level 2), respectively. These results clearly demonstrated that the acidic solvent, which combined oil extraction with in situ transesterification, was an effective catalyst for the production of high-quantity, high-quality biodiesel from a Scenedesmus sp. PMID:24689039

  19. [Development of the Chlamydomonas actinochloris culture after microwave irradiation].

    PubMed

    Grigor'eva, O O; Berezovskaia, M A; Datsenko, A I

    2012-01-01

    Effect of the microwave irradiation on the subsequent development of the Chlamydomonas actinochloris culture is studied. The number of cells in the suspension was controlled and photoluminescence measurements were performed for 25 days to estimate the functional state of the cells. The exposure at a dose of 80 J/g is shown to negligibly affect the green alga, whereas the 122 J/g dose led to deterioration of the functional state and, thereafter, to the death of most cells. However, the survivors intensively developed, the culture restored the normal state for 20 days, reached and later even left behind the control sample in development.

  20. Production of therapeutic proteins in the chloroplast of Chlamydomonas reinhardtii

    PubMed Central

    2014-01-01

    Chloroplast transformation in the photosynthetic alga Chlamydomonas reinhardtii has been used to explore the potential to use it as an inexpensive and easily scalable system for the production of therapeutic recombinant proteins. Diverse proteins, such as bacterial and viral antigens, antibodies and, immunotoxins have been successfully expressed in the chloroplast using endogenous and chimeric promoter sequences. In some cases, proteins have accumulated to high level, demonstrating that this technology could compete with current production platforms. This review focuses on the works that have engineered the chloroplast of C. reinhardtii with the aim of producing recombinant proteins intended for therapeutical use in humans or animals. PMID:25136510

  1. Production of therapeutic proteins in the chloroplast of Chlamydomonas reinhardtii.

    PubMed

    Almaraz-Delgado, Alma Lorena; Flores-Uribe, José; Pérez-España, Víctor Hugo; Salgado-Manjarrez, Edgar; Badillo-Corona, Jesús Agustín

    2014-01-01

    Chloroplast transformation in the photosynthetic alga Chlamydomonas reinhardtii has been used to explore the potential to use it as an inexpensive and easily scalable system for the production of therapeutic recombinant proteins. Diverse proteins, such as bacterial and viral antigens, antibodies and, immunotoxins have been successfully expressed in the chloroplast using endogenous and chimeric promoter sequences. In some cases, proteins have accumulated to high level, demonstrating that this technology could compete with current production platforms. This review focuses on the works that have engineered the chloroplast of C. reinhardtii with the aim of producing recombinant proteins intended for therapeutical use in humans or animals.

  2. Symbiodinium thermophilum sp. nov., a thermotolerant symbiotic alga prevalent in corals of the world's hottest sea, the Persian/Arabian Gulf.

    PubMed

    Hume, B C C; D'Angelo, C; Smith, E G; Stevens, J R; Burt, J; Wiedenmann, J

    2015-02-27

    Coral reefs are in rapid decline on a global scale due to human activities and a changing climate. Shallow water reefs depend on the obligatory symbiosis between the habitat forming coral host and its algal symbiont from the genus Symbiodinium (zooxanthellae). This association is highly sensitive to thermal perturbations and temperatures as little as 1°C above the average summer maxima can cause the breakdown of this symbiosis, termed coral bleaching. Predicting the capacity of corals to survive the expected increase in seawater temperatures depends strongly on our understanding of the thermal tolerance of the symbiotic algae. Here we use molecular phylogenetic analysis of four genetic markers to describe Symbiodinium thermophilum, sp. nov. from the Persian/Arabian Gulf, a thermally tolerant coral symbiont. Phylogenetic inference using the non-coding region of the chloroplast psbA gene resolves S. thermophilum as a monophyletic lineage with large genetic distances from any other ITS2 C3 type found outside the Gulf. Through the characterisation of Symbiodinium associations of 6 species (5 genera) of Gulf corals, we demonstrate that S. thermophilum is the prevalent symbiont all year round in the world's hottest sea, the southern Persian/Arabian Gulf.

  3. The production of sulfonated chitosan-sodium alginate found in brown algae (Sargassum sp.) composite membrane as proton exchange membrane fuel cell (PEMFC)

    NASA Astrophysics Data System (ADS)

    Wafiroh, Siti; Pudjiastuti, Pratiwi; Sari, Ilma Indana

    2016-03-01

    The majority of energy was used in this period is from fossil fuel, which getting decreased in the future. The objective of this research is production and characterization of sulfonated chitosan-sodium alginate found in brown algae (Sargassum sp.) composite membrane as Proton Exchange Membrane Fuel Cell (PEMFC) for alternative energy. PEMFC was produced with 4 variations (w/w) ratio between chitosan and sodium alginate, 8 : 0, 8 : 1, 8 : 2, 8 : 4 (w/w). The production of membrane was mixed sodium alginate solution into chitosan solution and sulfonated with H2SO4 0.72 N. The characterization of the PEM was uses Modulus Young analysis, water swelling, ion exchange capacity, FTIR, SEM, DTA, methanol permeability and proton conductivity. The result of the research, showed that the optimum membrane was with ratio 8 : 2 (w/w) that the Modulus Young 8564 kN/m2, water swelling 31.86%, ion exchange capacity 1.020 meq/g, proton conductivity 8,8 × 10-6 S/cm, methanol permeability 1.90 × 10-8 g/cm2s and glass transition temperature (Tg) 100.9 °C, crystalline temperature (Tc) 227.6 °C, and the melting temperature (Tm) 267.9 °C.

  4. Symbiodinium thermophilum sp. nov., a thermotolerant symbiotic alga prevalent in corals of the world's hottest sea, the Persian/Arabian Gulf

    PubMed Central

    Hume, B. C. C.; D'Angelo, C.; Smith, E. G.; Stevens, J. R.; Burt, J.; Wiedenmann, J.

    2015-01-01

    Coral reefs are in rapid decline on a global scale due to human activities and a changing climate. Shallow water reefs depend on the obligatory symbiosis between the habitat forming coral host and its algal symbiont from the genus Symbiodinium (zooxanthellae). This association is highly sensitive to thermal perturbations and temperatures as little as 1°C above the average summer maxima can cause the breakdown of this symbiosis, termed coral bleaching. Predicting the capacity of corals to survive the expected increase in seawater temperatures depends strongly on our understanding of the thermal tolerance of the symbiotic algae. Here we use molecular phylogenetic analysis of four genetic markers to describe Symbiodinium thermophilum, sp. nov. from the Persian/Arabian Gulf, a thermally tolerant coral symbiont. Phylogenetic inference using the non-coding region of the chloroplast psbA gene resolves S. thermophilum as a monophyletic lineage with large genetic distances from any other ITS2 C3 type found outside the Gulf. Through the characterisation of Symbiodinium associations of 6 species (5 genera) of Gulf corals, we demonstrate that S. thermophilum is the prevalent symbiont all year round in the world's hottest sea, the southern Persian/Arabian Gulf. PMID:25720577

  5. Activation of Autophagy by Metals in Chlamydomonas reinhardtii.

    PubMed

    Pérez-Martín, Marta; Blaby-Haas, Crysten E; Pérez-Pérez, María Esther; Andrés-Garrido, Ascensión; Blaby, Ian K; Merchant, Sabeeha S; Crespo, José L

    2015-09-01

    Autophagy is an intracellular self-degradation pathway by which eukaryotic cells recycle their own material in response to specific stress conditions. Exposure to high concentrations of metals causes cell damage, although the effect of metal stress on autophagy has not been explored in photosynthetic organisms. In this study, we investigated the effect of metal excess on autophagy in the model unicellular green alga Chlamydomonas reinhardtii. We show in cells treated with nickel an upregulation of ATG8 that is independent of CRR1, a global regulator of copper signaling in Chlamydomonas. A similar effect on ATG8 was observed with copper and cobalt but not with cadmium or mercury ions. Transcriptome sequencing data revealed an increase in the abundance of the protein degradation machinery, including that responsible for autophagy, and a substantial overlap of that increased abundance with the hydrogen peroxide response in cells treated with nickel ions. Thus, our results indicate that metal stress triggers autophagy in Chlamydomonas and suggest that excess nickel may cause oxidative damage, which in turn activates degradative pathways, including autophagy, to clear impaired components and recover cellular homeostasis.

  6. Phylogenomic analysis of the Chlamydomonas genome unmasks proteins potentially involved in photosynthetic function and regulation

    PubMed Central

    Karpowicz, Steven J.; Heinnickel, Mark; Dewez, David; Hamel, Blaise; Dent, Rachel; Niyogi, Krishna K.; Johnson, Xenie; Alric, Jean; Wollman, Francis-André; Li, Huiying; Merchant, Sabeeha S.

    2010-01-01

    Chlamydomonas reinhardtii, a unicellular green alga, has been exploited as a reference organism for identifying proteins and activities associated with the photosynthetic apparatus and the functioning of chloroplasts. Recently, the full genome sequence of Chlamydomonas was generated and a set of gene models, representing all genes on the genome, was developed. Using these gene models, and gene models developed for the genomes of other organisms, a phylogenomic, comparative analysis was performed to identify proteins encoded on the Chlamydomonas genome which were likely involved in chloroplast functions (or specifically associated with the green algal lineage); this set of proteins has been designated the GreenCut. Further analyses of those GreenCut proteins with uncharacterized functions and the generation of mutant strains aberrant for these proteins are beginning to unmask new layers of functionality/regulation that are integrated into the workings of the photosynthetic apparatus. PMID:20490922

  7. The Chlamydomonas Genome Reveals the Evolution of Key Animal and Plant Functions

    SciTech Connect

    Merchant, Sabeeha S

    2007-04-09

    Chlamydomonas reinhardtii is a unicellular green alga whose lineage diverged from land plants over 1 billion years ago. It is a model system for studying chloroplast-based photosynthesis, as well as the structure, assembly, and function of eukaryotic flagella (cilia), which were inherited from the common ancestor of plants and animals, but lost in land plants. We sequenced the 120-megabase nuclear genome of Chlamydomonas and performed comparative phylogenomic analyses, identifying genes encoding uncharacterized proteins that are likely associated with the function and biogenesis of chloroplasts or eukaryotic flagella. Analyses of the Chlamydomonas genome advance our understanding of the ancestral eukaryotic cell, reveal previously unknown genes associated with photosynthetic and flagellar functions, and establish links between ciliopathy and the composition and function of flagella.

  8. Exploring the electron transfer pathways in photosystem I by high-time-resolution electron paramagnetic resonance: observation of the B-side radical pair P700(+)A1B(-) in whole cells of the deuterated green alga Chlamydomonas reinhardtii at cryogenic temperatures.

    PubMed

    Berthold, Thomas; von Gromoff, Erika Donner; Santabarbara, Stefano; Stehle, Patricia; Link, Gerhard; Poluektov, Oleg G; Heathcote, Peter; Beck, Christoph F; Thurnauer, Marion C; Kothe, Gerd

    2012-03-28

    Crystallographic models of photosystem I (PS I) highlight a symmetrical arrangement of the electron transfer cofactors which are organized in two parallel branches (A, B) relative to a pseudo-C2 symmetry axis that is perpendicular to the membrane plane. Here, we explore the electron transfer pathways of PS I in whole cells of the deuterated green alga Chlamydomonas reinhardtii using high-time-resolution electron paramagnetic resonance (EPR) at cryogenic temperatures. Particular emphasis is given to quantum oscillations detectable in the tertiary radical pairs P700(+)A1A(-) and P700(+)A1B(-) of the electron transfer chain. Results are presented first for the deuterated site-directed mutant PsaA-M684H in which electron transfer beyond the primary electron acceptor A0A on the PsaA branch of electron transfer is impaired. Analysis of the quantum oscillations, observed in a two-dimensional Q-band (34 GHz) EPR experiment, provides the geometry of the B-side radical pair. The orientation of the g tensor of P700(+) in an external reference system is adapted from a time-resolved multifrequency EPR study of deuterated and 15N-substituted cyanobacteria (Link, G.; Berthold, T.; Bechtold, M.; Weidner, J.-U.; Ohmes, E.; Tang, J.; Poluektov, O.; Utschig, L.; Schlesselman, S. L.; Thurnauer, M. C.; Kothe, G. J. Am. Chem. Soc. 2001, 123, 4211-4222). Thus, we obtain the three-dimensional structure of the B-side radical pair following photoexcitation of PS I in its native membrane. The new structure describes the position and orientation of the reduced B-side quinone A1B(-) on a nanosecond time scale after light-induced charge separation. Furthermore, we present results for deuterated wild-type cells of C. reinhardtii demonstrating that both radical pairs P700(+)A1A(-) and P700(+)A1B(-) participate in the electron transfer process according to a mole ratio of 0.71/0.29 in favor of P700(+)A1A(-). A detailed comparison reveals different orientations of A1A(-) and A1B(-) in their

  9. Larvicidal algae.

    PubMed

    Marten, Gerald G

    2007-01-01

    Although most algae are nutritious food for mosquito larvae, some species kill the larvae when ingested in large quantities. Cyanobacteria (blue-green algae) that kill larvae do so by virtue of toxicity. While blue-green algae toxins may offer possibilities for delivery as larvicides, the toxicity of live blue-green algae does not seem consistent enough for live algae to be useful for mosquito control. Certain species of green algae in the order Chlorococcales kill larvae primarily because they are indigestible. Where these algae are abundant in nature, larvae consume them to the exclusion of other food and then starve. Under the right circumstances, it is possible to introduce indigestible algae into a breeding habitat so they become abundant enough to render it unsuitable for mosquito production. The algae can persist for years, even if the habitat dries periodically. The main limitation of indigestible algae lies in the fact that, under certain conditions, they may not replace all the nutritious algae in the habitat. More research on techniques to ensure complete replacement will be necessary before indigestible algae can go into operational use for mosquito control.

  10. Rapid induction of lipid droplets in Chlamydomonas reinhardtii and Chlorella vulgaris by Brefeldin A.

    PubMed

    Kim, Sangwoo; Kim, Hanul; Ko, Donghwi; Yamaoka, Yasuyo; Otsuru, Masumi; Kawai-Yamada, Maki; Ishikawa, Toshiki; Oh, Hee-Mock; Nishida, Ikuo; Li-Beisson, Yonghua; Lee, Youngsook

    2013-01-01

    Algal lipids are the focus of intensive research because they are potential sources of biodiesel. However, most algae produce neutral lipids only under stress conditions. Here, we report that treatment with Brefeldin A (BFA), a chemical inducer of ER stress, rapidly triggers lipid droplet (LD) formation in two different microalgal species, Chlamydomonas reinhardtii and Chlorella vulgaris. LD staining using Nile red revealed that BFA-treated algal cells exhibited many more fluorescent bodies than control cells. Lipid analyses based on thin layer chromatography and gas chromatography revealed that the additional lipids formed upon BFA treatment were mainly triacylglycerols (TAGs). The increase in TAG accumulation was accompanied by a decrease in the betaine lipid diacylglyceryl N,N,N-trimethylhomoserine (DGTS), a major component of the extraplastidic membrane lipids in Chlamydomonas, suggesting that at least some of the TAGs were assembled from the degradation products of membrane lipids. Interestingly, BFA induced TAG accumulation in the Chlamydomonas cells regardless of the presence or absence of an acetate or nitrogen source in the medium. This effect of BFA in Chlamydomonas cells seems to be due to BFA-induced ER stress, as supported by the induction of three homologs of ER stress marker genes by the drug. Together, these results suggest that ER stress rapidly triggers TAG accumulation in two green microalgae, C. reinhardtii and C. vulgaris. A further investigation of the link between ER stress and TAG synthesis may yield an efficient means of producing biofuel from algae.

  11. Effective viscosity of non-gravitactic Chlamydomonas Reinhardtii microswimmer suspensions

    NASA Astrophysics Data System (ADS)

    Mussler, Matthias; Rafaï, Salima; Peyla, Philippe; Wagner, Christian

    2013-03-01

    Active microswimmers are known to affect the macroscopic viscosity of suspensions in a more complex manner than passive particles. For puller-like microswimmers an increase in the viscosity has been observed. It has been suggested that the persistence of the orientation of the microswimmers hinders the rotation that is normally caused by the vorticity. It was previously shown that some sorts of algae are bottom-heavy swimmers, i.e., their centre of mass is not located in the centre of the body. In this way, the algae affect the vorticity of the flow when they are perpendicularly oriented to the axis of gravity. This orientation of gravity to vorticity is given in a rheometer that is equipped with a cone-plate geometry. Here we present measurements of the viscosity both in a cone-plate and a Taylor-Couette cell. The two set-ups yielded the same increase in viscosity although the axis of gravitation in the Taylor-Couette cell is parallel to the direction of vorticity. In a complementary experiment we tested the orientation of the direction of swimming through microscopic observation of single Chlamydomonas reinhardtii and could not identify a preferred orientation, i.e., our specific strain of Chlamydomonas reinhardtii are not bottom-heavy swimmers. We thus conclude that bottom heaviness is not a prerequisite for the increase of viscosity and that the effect of gravity on the rheology of our strain of Chlamydomonas reinhardtii is negligible. This finding reopens the question of whether the origin of persistence in the orientation of cells is actually responsible for the increased viscosity of the suspension.

  12. Polaribacter porphyrae sp. nov., isolated from the red alga Porphyra yezoensis, and emended descriptions of the genus Polaribacter and two Polaribacter species.

    PubMed

    Fukui, Youhei; Abe, Mahiko; Kobayashi, Masahiro; Saito, Hiroaki; Oikawa, Hiroshi; Yano, Yutaka; Satomi, Masataka

    2013-05-01

    Three Gram-negative, non-motile, strictly aerobic strains, designated LNM-20(T), LCM-1 and LAM-13, were isolated from thalli of the marine red alga Porphyra yezoensis. Phylogenetic analysis based on 16S rRNA gene sequences revealed that the isolates were associated with the genus Polaribacter in the family Flavobacteriaceae and were most closely related to Polaribacter dokdonensis DSW-5(T) (96.2 % 16S rRNA gene sequence similarity) and Polaribacter gangjinensis K17-16(T) (95.0 %). The DNA G+C content of the isolates was 28.6-29.2 mol%. DNA-DNA hybridization analysis showed that the isolates belonged to a single species distinct from both of their closest relatives. The only isoprenoid quinone detected was menaquinone-6. The main polar lipids were phosphatidylethanolamine, two unidentified aminolipids and two unidentified lipids. The major fatty acids were iso-C15 : 0, iso-C15 : 1ω10c and iso-C15 : 0 3-OH. The phenotypic features of strain LNM-20(T) differed from those of their closest relatives in several regards (colony colour, growth with 1 % NaCl and on TSA plus 2.5 % NaCl, hydrolysis of Tweens 40 and 80, and oxidization of five carbon compounds). On the basis of phylogenetic, chemotaxonomic and phenotypic analysis, the isolates represent a novel species in the genus Polaribacter, for which the name Polaribacter porphyrae sp. nov. is proposed. The type strain is LNM-20(T) ( = LMG 26671(T)  = NBRC 108759(T)). Emended descriptions of the genus Polaribacter and P. dokdonensis and P. gangjinensis are also proposed.

  13. In Polytomella sp. mitochondria, biogenesis of the heterodimeric COX2 subunit of cytochrome c oxidase requires two different import pathways.

    PubMed

    Jiménez-Suárez, Alejandra; Vázquez-Acevedo, Miriam; Rojas-Hernández, Andrés; Funes, Soledad; Uribe-Carvajal, Salvador; González-Halphen, Diego

    2012-05-01

    In the vast majority of eukaryotic organisms, the mitochondrial cox2 gene encodes subunit II of cytochrome c oxidase (COX2). However, in some lineages including legumes and chlorophycean algae, the cox2 gene migrated to the nucleus. Furthermore, in chlorophycean algae, this gene was split in two different units. Thereby the COX2 subunit is encoded by two independent nuclear genes, cox2a and cox2b, and mitochondria have to import the cytosol-synthesized COX2A and COX2B subunits and assemble them into the cytochrome c oxidase complex. In the chlorophycean algae Chlamydomonas reinhardtii and Polytomella sp., the COX2A precursor exhibits a long (130-140 residues), cleavable mitochondrial targeting sequence (MTS). In contrast, COX2B lacks an MTS, suggesting that mitochondria use different mechanisms to import each subunit. Here, we explored the in vitro import processes of both, the Polytomella sp. COX2A precursor and the COX2B protein. We used isolated, import-competent mitochondria from this colorless alga. Our results suggest that COX2B is imported directly into the intermembrane space, while COX2A seems to follow an energy-dependent import pathway, through which it finally integrates into the inner mitochondrial membrane. In addition, the MTS of the COX2A precursor is eliminated. This is the first time that the in vitro import of split COX2 subunits into mitochondria has been achieved.

  14. Analysis of flagellar phosphoproteins from Chlamydomonas reinhardtii.

    PubMed

    Boesger, Jens; Wagner, Volker; Weisheit, Wolfram; Mittag, Maria

    2009-07-01

    Cilia and flagella are cell organelles that are highly conserved throughout evolution. For many years, the green biflagellate alga Chlamydomonas reinhardtii has served as a model for examination of the structure and function of its flagella, which are similar to certain mammalian cilia. Proteome analysis revealed the presence of several kinases and protein phosphatases in these organelles. Reversible protein phosphorylation can control ciliary beating, motility, signaling, length, and assembly. Despite the importance of this posttranslational modification, the identities of many ciliary phosphoproteins and knowledge about their in vivo phosphorylation sites are still missing. Here we used immobilized metal affinity chromatography to enrich phosphopeptides from purified flagella and analyzed them by mass spectrometry. One hundred forty-one phosphorylated peptides were identified, belonging to 32 flagellar proteins. Thereby, 126 in vivo phosphorylation sites were determined. The flagellar phosphoproteome includes different structural and motor proteins, kinases, proteins with protein interaction domains, and many proteins whose functions are still unknown. In several cases, a dynamic phosphorylation pattern and clustering of phosphorylation sites were found, indicating a complex physiological status and specific control by reversible protein phosphorylation in the flagellum.

  15. Low oxygen levels contribute to improve photohydrogen production in mixotrophic non-stressed Chlamydomonas cultures

    DOE PAGES

    Jurado-Oller, Jose Luis; Dubini, Alexandra; Galvan, Aurora; ...

    2015-09-17

    Currently, hydrogen fuel is derived mainly from fossil fuels, but there is an increasing interest in clean and sustainable technologies for hydrogen production. In this context, the ability of some photosynthetic microorganisms, particularly cyanobacteria and microalgae, to produce hydrogen is a promising alternative for renewable, clean-energy production. Among a diverse array of photosynthetic microorganisms able to produce hydrogen, the green algae Chlamydomonas reinhardtii is the model organism widely used to study hydrogen production. Furthermore, the well-known fact that acetate-containing medium enhances hydrogen production in this algae, little is known about the precise role of acetate during this process.

  16. Low oxygen levels contribute to improve photohydrogen production in mixotrophic non-stressed Chlamydomonas cultures

    SciTech Connect

    Jurado-Oller, Jose Luis; Dubini, Alexandra; Galvan, Aurora; Fernandez, Emilio; Gonzalez-Ballester, David

    2015-09-17

    Currently, hydrogen fuel is derived mainly from fossil fuels, but there is an increasing interest in clean and sustainable technologies for hydrogen production. In this context, the ability of some photosynthetic microorganisms, particularly cyanobacteria and microalgae, to produce hydrogen is a promising alternative for renewable, clean-energy production. Among a diverse array of photosynthetic microorganisms able to produce hydrogen, the green algae Chlamydomonas reinhardtii is the model organism widely used to study hydrogen production. Furthermore, the well-known fact that acetate-containing medium enhances hydrogen production in this algae, little is known about the precise role of acetate during this process.

  17. Roseitalea porphyridii gen. nov., sp. nov., isolated from a red alga and reclassification of Hoeflea suaedae (Chung et al., 2013) as Pseudohoeflea suaedae gen. nov., comb. nov.

    PubMed

    Hyeon, Jong Woo; Jeong, Sang Eun; Baek, Kyunghwa; Jeon, Che Ok

    2016-11-02

    A Gram-staining-negative, strictly aerobic bacterial strain, designated MA7-20T, was isolated from a marine alga, Porphyridium marinum, in Korea. Cells showing oxidase-positive and catalase-positive activities were motile rods with bipolar flagella. Growth of strain MA7-20T was observed at 15-45 C (optimum, 30-37 C), at pH 6.0-10.5 (optimum, pH 7.0-8.0) and in the presence of 0-7 % (w/v) NaCl (optimum, 2-3 %). Strain MA7-20T contained summed feature 8 (comprising C18:1 ω7c/C18:1 ω6c) and 11-methyl-C18:1 ω7c and C18:0 as the major fatty acids and ubiquinone-10 as the sole isoprenoid quinone. The major polar lipids were phosphatidylcholine, phosphatidylglycerol, diphosphatidylglycerol, phosphatidylethanolamine and phosphatidyl-N-methylethanolamine. The G+C content of the genomic DNA was 61.5 mol%. Strain MA7-20T was most closely related to Hoeflea suaedae YC6898T, Oricola cellulosilytica CC-AMH-OT and Nitratireductor basaltis J3T with 96.0 %, 95.8 % and 95.8 % 16S rRNA gene sequence similarities, respectively, but the strain formed a distinct phylogenetic lineage from them within the family Phyllobacteriaceae with a low bootstrap value. H. suaedae also formed a clearly distinct phylogenetic lineage from other members of the genus Hoeflea and closely related genera. On the basis of phenotypic, chemotaxonomic and molecular properties, strain MA7-20T represents a novel genus of the family Phyllobacteriaceae, for which the name Roseitalea porphyridii gen. nov., sp. nov. is proposed. The type strain is MA7-20T (=KACC 18807T =JCM 31538T). In addition, H. suaedae is also reclassified as Pseudohoeflea suaedae gen. nov., comb. nov. (type strain YC6898T =KACC 14911T =NBRC 107700T).

  18. Granulosicoccus undariae sp. nov., a member of the family Granulosicoccaceae isolated from a brown algae reservoir and emended description of the genus Granulosicoccus.

    PubMed

    Park, Sooyeon; Jung, Yong-Taek; Won, Sung-Min; Park, Ja-Min; Yoon, Jung-Hoon

    2014-11-01

    A Gram-stain-negative, aerobic, non-flagellated and coccoid bacterial strain, W-BA3(T), which was isolated from a brown algae reservoir in Wando of South Korea, was characterized taxonomically. Strain W-BA3(T) was found to grow optimally at 30 °C, at pH 7.0-8.0 and in presence of 2.0 % (w/v) NaCl. In the neighbour-joining phylogenetic tree based on 16S rRNA gene sequences, strain W-BA3(T) clustered with the type strains of Granulosicoccus antarcticus and Granulosicoccus coccoides, with which it exhibited sequence similarity values of 98.4-99.3 %. Sequence similarity values of strain W-BA3(T) to the type strains of the other recognized species were less than 90.2 %. Strain W-BA3(T) was found to contain Q-8 as the predominant ubiquinone and summed feature 3 (C16:1 ω7c and/or C16:1 ω6c), C18:1 ω7c and C16:0 as the major fatty acids. The major polar lipids of strain W-BA3(T), which were identified as phosphatidylethanolamine and phosphatidylglycerol, were similar to those of the type strains of G. antarcticus and G. coccoides. The DNA G+C content of strain W-BA3(T) was 56.0 mol % and its mean DNA-DNA relatedness values with the type strains of G. coccoides and G. antarcticus were 27 and 17 %, respectively. Differential phenotypic properties, together with the phylogenetic and genetic distinctiveness, demonstrated that strain W-BA3(T) is separated from the two Granulosicoccus species. On the basis of the data presented, strain W-BA3(T) is considered to represent a novel species of the genus Granulosicoccus, for which the name Granulosicoccus undariae sp. nov. is proposed. The type strain is W-BA3(T) (=KCTC 42134(T) = NBRC 110411(T)). An emended description of the genus Granulosicoccus is also proposed.

  19. Catenovulum maritimus sp. nov., a novel agarolytic gammaproteobacterium isolated from the marine alga Porphyra yezoensis Ueda (AST58-103), and emended description of the genus Catenovulum.

    PubMed

    Li, Dong-Qi; Zhou, Yan-Xia; Liu, Tao; Chen, Guan-Jun; Du, Zong-Jun

    2015-08-01

    A novel agarolytic, Gram-stain negative, heterotrophic, facultatively anaerobic and pale-white pigmented bacterial strain, designated Q1(T), was isolated from the marine alga Porphyra yezoensis Ueda (AST58-103) collected from the coastal area of Weihai, China. The cells are motile by means of peritrichous flagella. The isolate requires NaCl for growth, while seawater is not necessary, and growth occurs optimally at about 30-33 °C, in 1-3 % (w/v) NaCl and at pH 7-7.5. Strain Q1(T) shows oxidase-positive and catalase-negative activities, and possesses the ability to hydrolyse starch and alginate, but not cellulose, gelatin, urea or Tween-80. Phylogenetic analysis based on 16S rRNA gene sequence indicated that strain Q1(T) is affiliated with the family Alteromonadaceae within the class Gammaproteobacteria. The isolate, strain Q1(T), is most closely related to Catenovulum agarivorans YM01(T) (94.85 %), with less than 91.2 % sequence similarity to other close relatives with validly published names. The draft genome sequence of strain Q1(T) consists of 62 contigs (>200 bp) of 4,548,270 bp. The genomes of Q1(T) and YM01(T) have an ANI value of 70.7 %, and the POCP value between the two genomes is 64.4 %. The genomic DNA G+C content of strain Q1(T) is 37.9 mol% as calculated from the draft genome sequence. The main isoprenoid quinone is ubiquinone-8. The predominant cellular fatty acids are summed feature 3 (C16:1 ω7c and/or iso-C15:0 2-OH), C16:0 and C18:1 ω7c. The major polar lipids are phosphatidylethanolamine and phosphatidylglycerol. Based on data from a polyphasic chemotaxonomic, physiological and biochemical study, strain Q1(T) should be classified as a novel species of the genus Catenovulum, for which the name Catenovulum maritimus sp. nov. is proposed. The type strain is Q1(T) (=CICC 10836(T)=DSM 28813(T)).

  20. 13th International Conference on Chlamydomonas

    SciTech Connect

    Silflow, Carolyn D.

    2014-03-11

    The 13th International Conference on Chlamydomonas (EMBO Workshop on the Cell and Molecular Biology of Chlamydomonas) was held May 27 to June 1, 2008 in Hyeres, France. The conference was the biennial meeting for all researchers studying the green algal systems Chlamydomonas and Volvox. The conference brought together approximately 200 investigators from around the world (North America, Asia, Europe and Australia) representing different fields and disciplines (cell biology, genetics, biochemistry, biophysics, plant physiology, genomics). It provided an opportunity for investigators from different countries to share methodologies and to discuss recent results with a focus on the Chlamydomonas experimental system.

  1. High-fidelity phototaxis in biflagellate algae

    NASA Astrophysics Data System (ADS)

    Leptos, Kyriacos; Chioccioli, Maurizio; Furlan, Silvano; Pesci, Adriana; Goldstein, Raymond

    2015-11-01

    The single-cell alga Chlamydomonas reinhardtii is a motile biflagellate that can swim towards light for its photosynthetic requirements, a behavior referred to as phototaxis. The cell responds upon light stimulation through its rudimentary eye - the eyespot - by changing the beating amplitude of its two flagella accordingly - a process called the photoresponse. All this occurs in a coordinated fashion as Chlamydomonas spins about its body axis while swimming, thus experiencing oscillating intensities of light. We use high-speed video microscopy to measure the flagellar dynamics of the photoresponse on immobilized cells and interpret the results with a mathematical model of adaptation similar to that used previously for Volvox. These results are incorporated into a model of phototactic steering to yield trajectories that are compared to those obtained by three-dimensional tracking. Implications of these results for the evolution of multicellularity in the Volvocales are discussed.

  2. Enhanced accumulation of PCB congeners by Baltic Sea blue mussels, Mytilus edulis, with increased algae enrichment

    SciTech Connect

    Gilek, M.; Bjoerk, M.; Broman, D.; Kautsky, N.; Naef, C.

    1996-09-01

    The objective of this study was to examine if natural variations in the quantity of phytoplankton-derived particulate and dissolved organic carbon influences the accumulation of polychlorinated biphenyls (PCBs) in the tissues of Baltic Sea blue mussels (Mytilus edulis L.). In a laboratory flow-through experiment the authors exposed M. edulis to the technical PCB mixture Aroclor{reg_sign} 1248 for 21 d at three different enrichments of the unicellular green algae Chlamydomonas sp., 0.10, 0.16, and 0.32 mg particulate organic carbon (POC)/L. Tissue and water concentrations were determined for seven PCB congeners and 21-d bioaccumulation factors were calculated against total water concentrations. Contrary to what would be expected, an increase in algae enrichment from 0.10 to 0.32 mg POC/L resulted in an enhanced PCB accumulation by a factor of approx. 2. This increase in PCB accumulation was more pronounced for PCB congeners with lower hydrophobicity. These observations have implications for the design of laboratory accumulation studies and potentially for PCB accumulation and cycling in field populations of suspension-feeding mussels in response to changes in eutrophication status.

  3. The Study of Algae

    ERIC Educational Resources Information Center

    Rushforth, Samuel R.

    1977-01-01

    Included in this introduction to the study of algae are drawings of commonly encountered freshwater algae, a summary of the importance of algae, descriptions of the seven major groups of algae, and techniques for collection and study of algae. (CS)

  4. Individual Flagellar Waveform Affects Collective Behavior of Chlamydomonas reinhardtii.

    PubMed

    Kage, Azusa; Mogami, Yoshihiro

    2015-08-01

    Bioconvection is a form of collective motion that occurs spontaneously in the suspension of swimming microorganisms. In a previous study, we quantitatively described the "pattern transition," a phase transition phenomenon that so far has exclusively been observed in bioconvection of the unicellular green alga Chlamydomonas. We suggested that the transition could be induced by changes in the balance between the gravitational and shear-induced torques, both of which act to determine the orientation of the organism in the shear flow. As both of the torques should be affected by the geometry of the Chlamydomonas cell, alteration in the flagellar waveform might change the extent of torque generation by altering overall geometry of the cell. Based on this working hypothesis, we examined bioconvection behavior of two flagellar mutants of Chlamydomonas reinhardtii, ida1 and oda2, making reference to the wild type. Flagella of ida1 beat with an abnormal waveform, while flagella of oda2 show a normal waveform but lower beat frequency. As a result, both mutants had swimming speed of less than 50% of the wild type. ida1 formed bioconvection patterns with smaller spacing than those of wild type and oda2. Two-axis view revealed the periodic movement of the settling blobs of ida1, while oda2 showed qualitatively similar behavior to that of wild type. Unexpectedly, ida1 showed stronger negative gravitaxis than did wild type, while oda2 showed relatively weak gravitaxis. These findings suggest that flagellar waveform, not swimming speed or beat frequency, strongly affect bioconvection behavior in C. reinhardtii.

  5. New insights into the roles of molecular chaperones in Chlamydomonas and Volvox.

    PubMed

    Nordhues, André; Miller, Stephen M; Mühlhaus, Timo; Schroda, Michael

    2010-01-01

    The unicellular green alga Chlamydomonas reinhardtii has been used as a model organism for many decades, mainly to study photosynthesis and flagella/cilia. Only recently, Chlamydomonas has received much attention because of its ability to produce hydrogen and nonpolar lipids that have promise as biofuels. The best-studied multicellular cousin of Chlamydomonas reinhardtii is Volvox carteri, whose life cycle comprises events that have clear parallels in higher plants and/or animals, making it an excellent system in which to study fundamental developmental processes. Molecular chaperones are proteins that guide other cellular proteins through their life cycle. They assist in de novo folding of nascent chains, mediate assembly and disassembly of protein complexes, facilitate protein transport across membranes, disassemble protein aggregates, fold denatured proteins back to the native state, and transfer unfoldable proteins to proteolytic degradation. Hence, molecular chaperones regulate protein function under all growth conditions and play important roles in many basic cellular and developmental processes. The aim of this chapter is to describe recent advances toward understanding molecular chaperone biology in Chlamydomonas and Volvox.

  6. Propulsive Forces on the Flagellum during Locomotion of Chlamydomonas reinhardtii

    PubMed Central

    Bayly, P.V.; Lewis, B.L.; Ranz, E.C.; Okamoto, R.J.; Pless, R.B.; Dutcher, S.K.

    2011-01-01

    The distributed propulsive forces exerted on the flagellum of the swimming alga Chlamydomonas reinhardtii by surrounding fluid were estimated from experimental image data. Images of uniflagellate mutant Chlamydomonas cells were obtained at 350 frames/s with 125-nm spatial resolution, and the motion of the cell body and the flagellum were analyzed in the context of low-Reynolds-number fluid mechanics. Wild-type uniflagellate cells, as well as uniflagellate cells lacking inner dynein arms (ida3) or outer dynein arms (oda2) were studied. Ida3 cells exhibit stunted flagellar waveforms, whereas oda2 cells beat with lower frequency. Image registration and sorting algorithms provided high-resolution estimates of the motion of the cell body, as well as detailed kinematics of the flagellum. The swimming cell was modeled as an ellipsoid in Stokes flow, propelled by viscous forces on the flagellum. The normal and tangential components of force on the flagellum (fN and fT) were related by resistive coefficients (CN and CT) to the corresponding components of velocity (VN and VT).The values of these coefficients were estimated by satisfying equilibrium requirements for force and torque on the cell. The estimated values of the resistive coefficients are consistent among all three genotypes and similar to theoretical predictions. PMID:21641317

  7. New thioredoxin targets in the unicellular photosynthetic eukaryote Chlamydomonas reinhardtii

    PubMed Central

    Lemaire, Stéphane D.; Guillon, Blanche; Le Maréchal, Pierre; Keryer, Eliane; Miginiac-Maslow, Myroslawa; Decottignies, Paulette

    2004-01-01

    Proteomics were used to identify the proteins from the eukaryotic unicellular green alga Chlamydomonas reinhardtii that can be reduced by thioredoxin. These proteins were retained specifically on a thioredoxin affinity column made of a monocysteinic thioredoxin mutant able to form mixed disulfides with its targets. Of a total of 55 identified targets, 29 had been found previously in higher plants or Synechocystis, but 26 were new targets. Biochemical tests were performed on three of them, showing a thioredoxin-dependent activation of isocitrate lyase and isopropylmalate dehydrogenase and a thioredoxin-dependent deactivation of catalase that is redox insensitive in Arabidopsis. In addition, we identified a Ran protein, a previously uncharacterized nuclear target in a photosynthetic organism. The metabolic and evolutionary implications of these findings are discussed. PMID:15123830

  8. Regulation of Photosynthetic Capacity in Chlamydomonas mundana 1

    PubMed Central

    Russell, George K.; Gibbs, Martin

    1966-01-01

    A regulatory system has been described in the obligately phototrophic green alga Chlamydomonas mundana. Cells grown in acetate media are unable to fix carbon dioxide in the light but carry out a photoassimilation of acetate to carbohydrate: cells cultured with carbon dioxide as the sole source of cellular carbon carry out typical green plant photosynthesis. The control appears to take place at the level of the reductive pentose phosphate cycle. The presence of sodium acetate in the medium strongly inhibits formation of ribulose-1.5-diphosphate carboxylase, ribulose-5-phosphate kinase, and one of the 2 fructose-1,6-diphosphate aldolase activities of the cell. Ribose-5-phosphate isomerase is present in higher activity in autotrophic cells. Changes in the levels of triose phosphate dehydrogenase were also noted. The total pigment content of the cell and the photosynthetic electron transport reactions are not altered under different conditions of growth. PMID:16656335

  9. New thioredoxin targets in the unicellular photosynthetic eukaryote Chlamydomonas reinhardtii.

    PubMed

    Lemaire, Stéphane D; Guillon, Blanche; Le Maréchal, Pierre; Keryer, Eliane; Miginiac-Maslow, Myroslawa; Decottignies, Paulette

    2004-05-11

    Proteomics were used to identify the proteins from the eukaryotic unicellular green alga Chlamydomonas reinhardtii that can be reduced by thioredoxin. These proteins were retained specifically on a thioredoxin affinity column made of a monocysteinic thioredoxin mutant able to form mixed disulfides with its targets. Of a total of 55 identified targets, 29 had been found previously in higher plants or Synechocystis, but 26 were new targets. Biochemical tests were performed on three of them, showing a thioredoxin-dependent activation of isocitrate lyase and isopropylmalate dehydrogenase and a thioredoxin-dependent deactivation of catalase that is redox insensitive in Arabidopsis. In addition, we identified a Ran protein, a previously uncharacterized nuclear target in a photosynthetic organism. The metabolic and evolutionary implications of these findings are discussed.

  10. Reduction of PII signaling protein enhances lipid body production in Chlamydomonas reinhardtii.

    PubMed

    Zalutskaya, Zhanneta; Kharatyan, Nina; Forchhammer, Karl; Ermilova, Elena

    2015-11-01

    In all examined organisms that have the PII signal transduction machinery, PII coordinates the central C/N anabolic metabolism. In green algae and land plants, PII is localized in the chloroplast and controls the L-arginine biosynthetic pathway pathway. To elucidate additional functions of PII in the model photosynthetic organism Chlamydomonas reinhardtii (CrPII), we generated and analyzed four strains, in which PII was strongly under-expressed by artificial microRNA (GLB1-amiRNA strains). In response to nitrogen deficiency, Chlamydomonas produces triacylglycerols (TAGs) that are accumulated in lipid bodies (LB). Quantification of LBs by confocal microscopy in four GLB1-amiRNA strains showed that reduced PII levels resulted in over-accumulation of LBs compared to their parental strains. Moreover, knock-down of PII caused also an increase in the total TAG level. We propose that the larger yields of TAG-filled LBs in N-starved GLB1-amiRNA cells can be attributed to the strain's depleted PII level and their inability to properly control acetyl-CoA carboxylase activity (ACCase). Together, our results imply that PII in Chlamydomonas negatively controls TAG accumulation in LBs during acclimation to nitrogen starvation of the alga.

  11. Isodityrosine cross-linking mediates insolubilization of cell walls in Chlamydomonas.

    PubMed

    Waffenschmidt, S; Woessner, J P; Beer, K; Goodenough, U W

    1993-07-01

    Enzymatic removal of the cell wall induces vegetative Chlamydomonas reinhardtii cells to transcribe wall genes and synthesize new hydroxyproline-rich glycoproteins (HRGPs) related to the extensins found in higher plant cell walls. A cDNA expression library made from such induced cells was screened with antibodies to an oligopeptide containing the (SP)x repetitive domains found in Chlamydomonas wall proteins. One of the selected cDNAs encodes an (SP)x-rich polypeptide that also displays a repeated YGG motif. Ascorbate, a peroxidase inhibitor, and tyrosine derivatives were shown to inhibit insolubilization of both the vegetative and zygotic cell walls of Chlamydomonas, suggesting that oxidative cross-linking of tyrosines is occurring. Moreover, insolubilization of both walls was concomitant with a burst in H2O2 production and in extracellular peroxidase activity. Finally, both isodityrosine and dityrosine were found in hydrolysates of the insolubilized vegetative wall layer. We propose that the formation of tyrosine cross-links is essential to Chlamydomonas HRGP insolubilization.

  12. Isodityrosine cross-linking mediates insolubilization of cell walls in Chlamydomonas.

    PubMed Central

    Waffenschmidt, S; Woessner, J P; Beer, K; Goodenough, U W

    1993-01-01

    Enzymatic removal of the cell wall induces vegetative Chlamydomonas reinhardtii cells to transcribe wall genes and synthesize new hydroxyproline-rich glycoproteins (HRGPs) related to the extensins found in higher plant cell walls. A cDNA expression library made from such induced cells was screened with antibodies to an oligopeptide containing the (SP)x repetitive domains found in Chlamydomonas wall proteins. One of the selected cDNAs encodes an (SP)x-rich polypeptide that also displays a repeated YGG motif. Ascorbate, a peroxidase inhibitor, and tyrosine derivatives were shown to inhibit insolubilization of both the vegetative and zygotic cell walls of Chlamydomonas, suggesting that oxidative cross-linking of tyrosines is occurring. Moreover, insolubilization of both walls was concomitant with a burst in H2O2 production and in extracellular peroxidase activity. Finally, both isodityrosine and dityrosine were found in hydrolysates of the insolubilized vegetative wall layer. We propose that the formation of tyrosine cross-links is essential to Chlamydomonas HRGP insolubilization. PMID:7689882

  13. Developing molecular tools for Chlamydomonas reinhardtii

    NASA Astrophysics Data System (ADS)

    Noor-Mohammadi, Samaneh

    Microalgae have garnered increasing interest over the years for their ability to produce compounds ranging from biofuels to neutraceuticals. A main focus of researchers has been to use microalgae as a natural bioreactor for the production of valuable and complex compounds. Recombinant protein expression in the chloroplasts of green algae has recently become more routine; however, the heterologous expression of multiple proteins or complete biosynthetic pathways remains a significant challenge. To take full advantage of these organisms' natural abilities, sophisticated molecular tools are needed to be able to introduce and functionally express multiple gene biosynthetic pathways in its genome. To achieve the above objective, we have sought to establish a method to construct, integrate and express multigene operons in the chloroplast and nuclear genome of the model microalgae Chlamydomonas reinhardtii. Here we show that a modified DNA Assembler approach can be used to rapidly assemble multiple-gene biosynthetic pathways in yeast and then integrate these assembled pathways at a site-specific location in the chloroplast, or by random integration in the nuclear genome of C. reinhardtii. As a proof of concept, this method was used to successfully integrate and functionally express up to three reporter proteins (AphA6, AadA, and GFP) in the chloroplast of C. reinhardtii and up to three reporter proteins (Ble, AphVIII, and GFP) in its nuclear genome. An analysis of the relative gene expression of the engineered strains showed significant differences in the mRNA expression levels of the reporter genes and thus highlights the importance of proper promoter/untranslated-region selection when constructing a target pathway. In addition, this work focuses on expressing the cofactor regeneration enzyme phosphite dehydrogenase (PTDH) in the chloroplast and nuclear genomes of C. reinhardtii. The PTDH enzyme converts phosphite into phosphate and NAD(P)+ into NAD(P)H. The reduced

  14. Evolution of sex and mating loci: an expanded view from Volvocine algae.

    PubMed

    Umen, James G

    2011-12-01

    Sexual reproduction in Volvocine algae coevolved with the acquisition of multicellularity. Unicellular genera such as Chlamydomonas and small colonial genera from this group have classical mating types with equal-sized gametes, while larger multicellular genera such as Volvox have differentiated males and females that produce sperm and eggs respectively. Newly available sequence from the Volvox and Chlamydomonas genomes and mating loci open up the potential to investigate how sex-determining regions co-evolve with major changes in development and sexual reproduction. The expanded size and sequence divergence between the male and female haplotypes of the Volvox mating locus (MT) not only provide insights into how the colonial Volvocine algae might have evolved sexual dimorphism, but also raise questions about why the putative ancestral-like MT locus in Chlamydomonas shows less divergence between haplotypes than expected.

  15. The altitudinal distribution of snow algae on an Alaska glacier (Gulkana Glacier in the Alaska Range)

    NASA Astrophysics Data System (ADS)

    Takeuchi, Nozomu

    2001-12-01

    The altitudinal distribution of a snow algal community was investigated on an Alaska glacier (Gulkana Glacier in the Alaska Range) from 1270 to 1770 m a.s.l.. Seven species of snow and ice algae (Chlorophyta and cyanobacteria) were observed on the glacier surface. These species were Chlamydomonas nivalis, Mesotaenium berggrenii, Ancylonema nordenskioldii, Cylindrocystis brébissonii, Raphidonema sp., and two Oscillatoriaceae cyanobacteria. The altitudinal distribution of snow algae was different among the species: Cd. nivalis was distributed on the middle to upper area, M. berggrenii; A. nordenskioldii, and one Oscillatoriaceae cyanobacterium on the middle to lower area; Raphidonema sp. on the middle area; and Cyl. brébissonii and one Oscillatoriaceae cyanobacterium on the lower area. The total cell concentration and the cell volume biomass of the snow algae ranged from 4·4 × 103 to 9·9 × 105 cells ml-1 and from 33 to 2211 µl m-2 respectively. The cell volume biomass changed with altitude; the biomass increased with altitude below 1600 m a.s.l., and decreased above 1600 m a.s.l. The community structure showed that algae is discussed in terms of the physical and chemical condition of the glacier surface, and is compared with that on a Himalayan glacier. A larger biomass in the snow area on the Alaska glacier than that of the Himalayan glacier is likely due to less frequent snow cover in summer in Alaska. Small amounts of filamentous cyanobacteria on the Alaska glacier may allow washouts of unicellular green algae by running melt water and may cause a different pattern of altitudinal distribution of algal biomass on the ice area from the Himalayan glacier

  16. Effect of Interactions Among Algae on Nitrogen Fixation by Blue-Green Algae (Cyanobacteria) in Flooded Soils

    PubMed Central

    Wilson, John T.; Greene, Sarah; Alexander, Martin

    1979-01-01

    Nitrogen fixation (C2H2 reduction) by algae in flooded soil was limited by interactions within the algal community. Nitrogen fixation by either indigenous algae or Tolypothrix tenuis was reduced severalfold by a dense suspension of the green alga Nephrocytium sp. Similarly, interactions between the nitrogen-fixing alga (cyanobacterium) Aulosira 68 and natural densities of indigenous algae limited nitrogen-fixing activity in one of two soils examined. This was demonstrated by developing a variant of Aulosira 68 that was resistant to the herbicide simetryne at concentrations that prevented development of indigenous algae. More nitrogen was fixed by the resistant variant in flooded soil containing herbicide than was fixed in herbicide-free soil by either the indigenous algae or indigenous algae plus the parent strain of Aulosira. Interference from indigenous algae may hamper the development of nitrogen-fixing algae introduced into rice fields in attempts to increase biological nitrogen fixation. PMID:16345463

  17. BiP links TOR signaling to ER stress in Chlamydomonas.

    PubMed

    Crespo, José L

    2012-02-01

    The highly conserved target of rapamycin (TOR) Ser/Thr kinase promotes protein synthesis under favorable growth conditions in all eukaryotes. Downregulation of TOR signaling in the model unicellular green alga Chlamydomonas reinhardtii has recently revealed a link between control of protein synthesis, endoplasmic reticulum (ER) stress and the reversible modification of the BiP chaperone by phosphorylation. Inhibition of protein synthesis by rapamycin or cycloheximide resulted in the phosphorylation of BiP on threonine residues while ER stress induced by tunicamycin or heat shock caused the fast dephosphorylation of the protein. Regulation of BiP function by phosphorylation/dephosphorylation events was proposed in early studies in mammalian cells although no connection to TOR signaling has been established so far. Here I will discuss about the coordinated regulation of BiP modification by TOR and ER stress signals in Chlamydomonas.

  18. Rescue of a paralyzed-flagella mutant of Chlamydomonas by transformation

    SciTech Connect

    Diener, D.R.; Curry, A.M.; Johnson, K.A.; Williams, B.D.; Rosenbaum, J.L. ); Lefebvre, P.A. ); Kindle, K.L. )

    1990-08-01

    The biflagellate alga Chlamydomonas has been used extensively in the genetic and biochemical analysis of flagellar assembly and motility. The authors have restored motility to a paralyzed-flagella mutant of Chlamydomonas by transforming with the corresponding wild-type gene. A nitrate reductase-deficient paralyzed-flagella strain, nit1-305 pf-14, carrying mutations in the genes for nitrate reductase and radial spoke protein 3, was transformed with wild-type copies of both genes. Two-thirds of the cells that survived nitrate selection also regained motility, indicating that they had been transformed with both the nitrate reductase and radial spoke protein 3 genes. Transformants typically contained multiple copies of both genes, genetically linked to each other, but not linked to the original mutant loci. Complementation of paralyzed-flagella mutants by transformation is a powerful tool for investigating flagellar assembly and function.

  19. Nutrient removal from horticultural wastewater by benthic filamentous algae Klebsormidium sp., Stigeoclonium spp. and their communities: From laboratory flask to outdoor Algal Turf Scrubber (ATS).

    PubMed

    Liu, Junzhuo; Danneels, Bram; Vanormelingen, Pieter; Vyverman, Wim

    2016-04-01

    Benthic filamentous algae have evident advantages in wastewater treatment over unicellular microalgae, including the ease in harvesting and resistance to predation. To assess the potentials of benthic filamentous algae in treating horticultural wastewater under natural conditions in Belgium, three strains and their mixture with naturally wastewater-borne microalgae were cultivated in 250 ml Erlenmeyer flasks in laboratory as well as in 1 m(2) scale outdoor Algal Turf Scrubber (ATS) with different flow rates. Stigeoclonium competed well with the natural wastewater-borne microalgae and contributed to most of the biomass production both in Erlenmeyer flasks and outdoor ATS at flow rates of 2-6 L min(-1) (water velocity 3-9 cm s(-1)), while Klebsormidium was not suitable for growing in horticultural wastewater under the tested conditions. Flow rate had great effects on biomass production and nitrogen removal, while phosphorus removal was less influenced by flow rate due to other mechanisms than assimilation by algae.

  20. Inorganic carbon limitation and mixotrophic growth in Chlamydomonas from an acidic mining lake.

    PubMed

    Tittel, Jörg; Bissinger, Vera; Gaedke, Ursula; Kamjunke, Norbert

    2005-06-01

    Plankton communities in acidic mining lakes (pH 2.5-3.3) are species-poor because they face extreme environmental conditions, e.g. 150mg l(-1) Fe2+ +Fe3+. We investigated the growth characteristics of the dominant pigmented species, the flagellate Chlamydomonas acidophila, in semi-continuous culture experiments under in situ conditions. The following hypotheses were tested: (1) Low inorganic carbon (IC) concentrations in the epilimnion (e.g. 0.3 mg l(-1)) arising from the low pH limit phototrophic growth (H-1); (2) the additional use of dissolved organic carbon (mixotrophy) leads to higher growth rates under IC-limitation (H-2), and (3) phagotrophy is not relevant (H-3). H-1 was supported as the culture experiments, in situ PAR and IC concentrations indicated that IC potentially limited phototrophic growth in the mixed surface layers. H-2 was also supported: mixotrophic growth always exceeded pure phototrophic growth even when photosynthesis was saturated. Dark growth in filtered lake water illuminated prior to inoculation provided evidence that Chlamydomonas was able to use the natural DOC. The alga did not grow on bacteria, thus confirming H-3. Chlamydomonas exhibited a remarkable resistance to starvation in the dark. The compensation light intensity (ca. 20 micromol photons m(-2) s(-1)) and the maximum phototrophic growth (1.50 d(-1)) fell within the range of algae from non-acidic waters. Overall, Chlamydomonas, a typical r-strategist in circum-neutral systems, showed characteristics of a K-strategist in the stable, acidic lake environment in achieving moderate growth rates and minimizing metabolic losses.

  1. Nuclear gene targeting in Chlamydomonas using engineered zinc-finger nucleases.

    PubMed

    Sizova, Irina; Greiner, Andre; Awasthi, Mayanka; Kateriya, Suneel; Hegemann, Peter

    2013-03-01

    The unicellular green alga Chlamydomonas reinhardtii is a versatile model for fundamental and biotechnological research. A wide range of tools for genetic manipulation have been developed for this alga, but specific modification of nuclear genes is still not routinely possible. Here, we present a nuclear gene targeting strategy for Chlamydomonas that is based on the application of zinc-finger nucleases (ZFNs). Our approach includes (i) design of gene-specific ZFNs using available online tools, (ii) evaluation of the designed ZFNs in a Chlamydomonas in situ model system, (iii) optimization of ZFN activity by modification of the nuclease domain, and (iv) application of the most suitable enzymes for mutagenesis of an endogenous gene. Initially, we designed a set of ZFNs to target the COP3 gene that encodes the light-activated ion channel channelrhodopsin-1. To evaluate the designed ZFNs, we constructed a model strain by inserting a non-functional aminoglycoside 3'-phosphotransferase VIII (aphVIII) selection marker interspaced with a short COP3 target sequence into the nuclear genome. Upon co-transformation of this recipient strain with the engineered ZFNs and an aphVIII DNA template, we were able to restore marker activity and select paromomycin-resistant (Pm-R) clones with expressing nucleases. Of these Pm-R clones, 1% also contained a modified COP3 locus. In cases where cells were co-transformed with a modified COP3 template, the COP3 locus was specifically modified by homologous recombination between COP3 and the supplied template DNA. We anticipate that this ZFN technology will be useful for studying the functions of individual genes in Chlamydomonas.

  2. UV-B Perception and Acclimation in Chlamydomonas reinhardtii[OPEN

    PubMed Central

    Chappuis, Richard; Allorent, Guillaume

    2016-01-01

    Plants perceive UV-B, an intrinsic component of sunlight, via a signaling pathway that is mediated by the photoreceptor UV RESISTANCE LOCUS8 (UVR8) and induces UV-B acclimation. To test whether similar UV-B perception mechanisms exist in the evolutionarily distant green alga Chlamydomonas reinhardtii, we identified Chlamydomonas orthologs of UVR8 and the key signaling factor CONSTITUTIVELY PHOTOMORPHOGENIC1 (COP1). Cr-UVR8 shares sequence and structural similarity to Arabidopsis thaliana UVR8, has conserved tryptophan residues for UV-B photoreception, monomerizes upon UV-B exposure, and interacts with Cr-COP1 in a UV-B-dependent manner. Moreover, Cr-UVR8 can interact with At-COP1 and complement the Arabidopsis uvr8 mutant, demonstrating that it is a functional UV-B photoreceptor. Chlamydomonas shows apparent UV-B acclimation in colony survival and photosynthetic efficiency assays. UV-B exposure, at low levels that induce acclimation, led to broad changes in the Chlamydomonas transcriptome, including in genes related to photosynthesis. Impaired UV-B-induced activation in the Cr-COP1 mutant hit1 indicates that UVR8-COP1 signaling induces transcriptome changes in response to UV-B. Also, hit1 mutants are impaired in UV-B acclimation. Chlamydomonas UV-B acclimation preserved the photosystem II core proteins D1 and D2 under UV-B stress, which mitigated UV-B-induced photoinhibition. These findings highlight the early evolution of UVR8 photoreceptor signaling in the green lineage to induce UV-B acclimation and protection. PMID:27020958

  3. Algae Resources

    SciTech Connect

    2016-06-01

    Algae are highly efficient at producing biomass, and they can be found all over the planet. Many use sunlight and nutrients to create biomass, which contain key components—including lipids, proteins, and carbohydrates— that can be converted and upgraded to a variety of biofuels and products. A functional algal biofuels production system requires resources such as suitable land and climate, sustainable management of water resources, a supplemental carbon dioxide (CO2) supply, and other nutrients (e.g., nitrogen and phosphorus). Algae can be an attractive feedstock for many locations in the United States because their diversity allows for highpotential biomass yields in a variety of climates and environments. Depending on the strain, algae can grow by using fresh, saline, or brackish water from surface water sources, groundwater, or seawater. Additionally, they can grow in water from second-use sources such as treated industrial wastewater; municipal, agricultural, or aquaculture wastewater; or produced water generated from oil and gas drilling operations.

  4. Cell and molecular biology of Chlamydomonas

    SciTech Connect

    Not Available

    1988-01-01

    This document contains only the abstracts of 92 presentations on the biology of Chlamydomonas. Topics include gene transformations, gene regulation, biosynthetic pathways, cell surfaces, circadian clocks, and the development and structure of the flagellar apparatus. (TEM)

  5. Characterizing the Anaerobic Response of Chlamydomonas reinhardtii by Quantitative Proteomics

    PubMed Central

    Terashima, Mia; Specht, Michael; Naumann, Bianca; Hippler, Michael

    2010-01-01

    The versatile metabolism of the green alga Chlamydomonas reinhardtii is reflected in its complex response to anaerobic conditions. The anaerobic response is also remarkable in the context of renewable energy because C. reinhardtii is able to produce hydrogen under anaerobic conditions. To identify proteins involved during anaerobic acclimation as well as to localize proteins and pathways to the powerhouses of the cell, chloroplasts and mitochondria from C. reinhardtii in aerobic and anaerobic (induced by 8 h of argon bubbling) conditions were isolated and analyzed using comparative proteomics. A total of 2315 proteins were identified. Further analysis based on spectral counting clearly localized 606 of these proteins to the chloroplast, including many proteins of the fermentative metabolism. Comparative quantitative analyses were performed with the chloroplast-localized proteins using stable isotopic labeling of amino acids ([13C6]arginine/[12C6]arginine in an arginine auxotrophic strain). The quantitative data confirmed proteins previously characterized as induced at the transcript level as well as identified several new proteins of unknown function induced under anaerobic conditions. These proteins of unknown function provide new candidates for further investigation, which could bring insights for the engineering of hydrogen-producing alga strains. PMID:20190198

  6. Select Acetophenones Modulate Flagellar Motility in Chlamydomonas

    PubMed Central

    Evans, Shakila K.; Pearce, Austin A.; Ibezim, Prudence K.; Primm, Todd P.; Gaillard, Anne R.

    2009-01-01

    Acetophenones were screened for activity against positive phototaxis of Chlamydomonas cells, a process that requires coordinated flagellar motility. The structure-activity relationships of a series of acetophenones are reported, including acetophenones that affect flagellar motility and cell viability. Notably, 4-methoxyacetophenone, 3,4-dimethoxyacetophenone, and 4-hydroxyacetophenone induced negative phototaxis in Chlamydomonas, suggesting interference with activity of flagellar proteins and control of flagellar dominance. PMID:20659114

  7. Glycolate Pathway in Algae 1

    PubMed Central

    Hess, J. L.; Tolbert, N. E.

    1967-01-01

    No glycolate oxidase activity could be detected by manometric, isotopic, or spectrophotometric techniques in cell extracts from 5 strains of algae grown in the light with CO2. However, NADH:glyoxylate reductase, phosphoglycolate phosphatase and isocitrate dehydrogenase were detected in the cell extracts. The serine formed by Chlorella or Chlamydomonas after 12 seconds of photosynthetic 14CO2 fixation contained 70 to 80% of its 14C in the carboxyl carbon. This distribution of label in serine was similar to that in phosphoglycerate from the same experiment. Thus, in algae serine is probably formed directly from phosphoglycerate. These results differ from those of higher plants which form uniformly labeled serine from glycolate in short time periods when phosphoglycerate is still carboxyl labeled. In glycolate formed by algae in 5 and 10 seconds of 14CO2 fixation, C2 was at least twice as radioactive as C1. A similar skewed labeling in C2 and C3 of 3-phosphoglycerate and serine suggests a common precursor for glycolate and 3-phosphoglycerate. Glycine formed by the algae, however, from the same experiments was uniformly labeled. Manganese deficient Chlorella incorporated only 2% of the total 14CO2 fixed in 10 minutes into glycolate, while in normal Chlorella 30% of the total 14C was found in glycolate. Manganese deficient Chlorella also accumulated more 14C in glycine and serine. Glycolate excretion by Chlorella was maximal in 10 mm bicarbonate and occurred only in the light, and was not influenced by the addition of glycolate. No time dependent uptake of significant amounts of either glycolate or phosphoglycolate was observed. When small amounts of glycolate-2-14C were fed to Chlorella or Scenedesmus, only 2 to 3% was metabolized after 30 to 60 minutes. The algae were not capable of significant glycolate metabolism as is the higher plant. The failure to detect glycolate oxidase, the low level glycolate-14C metabolism, and the formation of serine from phosphoglycerate

  8. [Effect of microwaves on Chlamydomonas actinochloris culture in the stationary phase of growth].

    PubMed

    Grigor'eva, O O; Berezovskaia, M A; Datsenko, A I

    2013-01-01

    Effects of the microwave radiation on the culture of Chlamydomonas actinochloris green flagellar alga in the stationary phase of growth are studied. After exposure to radiation at the maximum dose of 125 J/g, the cell functional state worsened but all the studied parameters were restored in 20 days and in the long run found to be even better than the control indices. The data are compared with the similar ones obtained earlier for the lag phase culture. The studied sample is found to be more resistant to the irradiation than the previous one.

  9. Phosphatase of Chlamydomonas reinhardi: biochemical and cytochemical approach with specific mutants.

    PubMed Central

    Matagne, R F; Loppes, R; Deltour, R

    1976-01-01

    The unicellular alga Chlamydomonas reinhardi produces two constitutive acid phosphatases and three depressible phosphatases (a neutral and two alkaline ones) that can utilize napthyl phosphate as a substrate. Specific mutants depressible phosphatase were used to investigate biochemical properties and the cytochemical localization of these enzymes. The two constitutive phosphatases show similar pH optima (about 5.0) and Km values (2 x 10(-3) to 3.3 x 10(-3) M) but differ in their heat sensitivity and affinity for glycerophosphate. Images PMID:4437

  10. Emergent Run-and-Tumble Behavior in a Simple Model of Chlamydomonas with Intrinsic Noise

    NASA Astrophysics Data System (ADS)

    Bennett, Rachel R.; Golestanian, Ramin

    2013-04-01

    Recent experiments on the green alga Chlamydomonas that swims using synchronized beating of a pair of flagella have revealed that it exhibits a run-and-tumble behavior similar to that of bacteria such as E. coli. Using a simple purely hydrodynamic model that incorporates a stroke cycle and an intrinsic Gaussian white noise, we show that a stochastic run-and-tumble behavior could emerge due to the nonlinearity of the combined synchronization-rotation-translation dynamics. Our study suggests that nonlinear mechanics could be a significant contributing factor to how the trajectories of the microorganism are selected.

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

    PubMed

    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.

  12. A phenotypic screening platform to identify small molecule modulators of Chlamydomonas reinhardtii growth, motility and photosynthesis

    PubMed Central

    2012-01-01

    Chemical biology, the interfacial discipline of using small molecules as probes to investigate biology, is a powerful approach of developing specific, rapidly acting tools that can be applied across organisms. The single-celled alga Chlamydomonas reinhardtii is an excellent model system because of its photosynthetic ability, cilia-related motility and simple genetics. We report the results of an automated fitness screen of 5,445 small molecules and subsequent assays on motility/phototaxis and photosynthesis. Cheminformatic analysis revealed active core structures and was used to construct a naïve Bayes model that successfully predicts algal bioactive compounds. PMID:23158586

  13. MicroRNAs in a multicellular green alga Volvox carteri.

    PubMed

    Li, Jingrui; Wu, Yang; Qi, Yijun

    2014-01-01

    microRNAs (miRNAs) have emerged as key components in the eukaryotic gene regulatory network. We and others have previously identified many miRNAs in a unicellular green alga, Chlamydomonas reinhardtii. To investigate whether miRNA-mediated gene regulation is a general mechanism in green algae and how miRNAs have been evolved in the green algal lineage, we examined small RNAs in Volvox carteri, a multicellular species in the same family with Chlamydomonas reinhardtii. We identified 174 miRNAs in Volvox, with many of them being highly enriched in gonidia or somatic cells. The targets of the miRNAs were predicted and many of them were subjected to miRNA-mediated cleavage in vivo, suggesting that miRNAs play regulatory roles in the biology of green algae. Our catalog of miRNAs and their targets provides a resource for further studies on the evolution, biological functions, and genomic properties of miRNAs in green algae.

  14. Sexual reproduction and sex determination in green algae.

    PubMed

    Sekimoto, Hiroyuki

    2017-02-10

    The sexual reproductive processes of some representative freshwater green algae are reviewed. Chlamydomonas reinhardtii is a unicellular volvocine alga having two mating types: mating type plus (mt(+)) and mating type minus (mt(-)), which are controlled by a single, complex mating-type locus. Sexual adhesion between the gametes is mediated by sex-specific agglutinin molecules on their flagellar membranes. Cell fusion is initiated by an adhesive interaction between the mt(+) and mt(-) mating structures, followed by localized membrane fusion. The loci of sex-limited genes and the conformation of sex-determining regions have been rearranged during the evolution of volvocine algae; however, the essential function of the sex-determining genes of the isogamous unicellular Chlamydomonas reinhardtii is conserved in the multicellular oogamous Volvox carteri. The sexual reproduction of the unicellular charophycean alga, Closterium peracerosum-strigosum-littorale complex, is also focused on here. The sexual reproductive processes of heterothallic strains are controlled by two multifunctional sex pheromones, PR-IP and PR-IP Inducer, which independently promote multiple steps in conjugation at the appropriate times through different induction mechanisms. The molecules involved in sexual reproduction and sex determination have also been characterized.

  15. Research for Developing Renewable Biofuels from Algae

    SciTech Connect

    Black, Paul N.

    2012-12-15

    Task A. Expansion of knowledge related to lipid production and secretion in algae A.1 Lipid biosynthesis in target algal species; Systems biology approaches are being used in combination with recent advances in Chlorella and Chlamydomonas genomics to address lipid accumulation in response to defined nutrient regimes. The UNL Algal Group continues screening additional species of Chlorella and other naturally occurring algae for those with optimal triglyceride production; Of the strains examined by the DOE's Aquatic Species Program, green algae, several species of Chlorella represent the largest group from which oleaginous candidates have been identified; A.1.1. Lipid profiling; Neutral lipid accumulation is routinely monitored by Nile red and BODIPY staining using high throughput strategies to screen for naturally occurring algae that accumulate triglyceride. These strategies complement those using spectrofluorometry to quantify lipid accumulation; Neutral lipid accumulation is routinely monitored by high performance thin-layer chromatography (HPTLC) and high performance liquid chromatography (HPLC) of lipid extracts in conjunction with; Carbon portioning experiments have been completed and the data currently are being analyzed and prepared for publication; Methods in the Black lab were developed to identify and quantify triacylglycerol (TAG), major membrane lipids [diacylglycerol trimethylhomoserine, phosphatidylethanolamine and chloroplast glycolipids], biosynthetic intermediates such as diacylglycerol, phosphatidic acid and lysophospholipids and different species of acyl-coenzyme A (acyl CoA).

  16. Mutants of Chlamydomonas: tools to study thylakoid membrane structure, function and biogenesis.

    PubMed

    de Vitry, C; Vallon, O

    1999-06-01

    The unicellular green alga Chlamydomonas reinhardtii is a model system for the study of photosynthesis and chloroplast biogenesis. C. reinhardtii has a photosynthesis apparatus similar to that of higher plants and it grows at rapid rate (generation time about 8 h). It is a facultative phototroph, which allows the isolation of mutants unable to perform photosynthesis and its sexual cycle allows a variety of genetic studies. Transformation of the nucleus and chloroplast genomes is easily performed. Gene transformation occurs mainly by homologous recombination in the chloroplast and heterologous recombination in the nucleus. Mutants are precious tools for studies of thylakoid membrane structure, photosynthetic function and assembly. Photosynthesis mutants affected in the biogenesis of a subunit of a protein complex usually lack the entire complex; this pleiotropic effect has been used in the identification of the other subunits, in the attribution of spectroscopic signals and also as a 'genetic cleaning' process which facilitates both protein complex purification, absorption spectroscopy studies or freeze-fracture analysis. The cytochrome b6f complex is not required for the growth of C. reinhardtii, unlike the case of photosynthetic prokaryotes in which the cytochrome complex is also part of the respiratory chain, and can be uniquely studied in Chlamydomonas by genetic approaches. We describe in greater detail the use of Chlamydomonas mutants in the study of this complex.

  17. Partial purification of the chloroplast ATP synthase from Chlamydomonas reinhardtii and the cloning and sequencing of a cDNA encoding the gamma subunit

    SciTech Connect

    Yu, L.M.

    1988-01-01

    The chloroplast ATP synthase was partially purified from the green alga Chlamydomonas reinhardtii by extracting membranes with deoxycholate and KCl, followed by centrifugation and ammonium sulfate fractionation of the supernatant. The enzyme assay involved the reconstitution of such fractions with bacteriorhodopsin and soybean phospholipids to form vesicles capable of light-dependent ({sup 32}P)-phosphate esterification. A cDNA for the gamma subunit from Chlamydomonas was isolated, expressed in vitro and sequenced. It contains the entire coding region for the gamma subunit precursor. A 35 amino acid long transit peptide resides at the NH{sub 2}-terminus of a 323 amino acid long mature peptide that is 77% similar to the spinach gamma subunit. Six cysteines were found; three were conserved in Chlamydomonas and spinach.

  18. Altered fermentative metabolism in Chlamydomonas reinhardtii mutants lacking pyruvate formate lyase and both pyruvate formate lyase and alcohol dehydrogenase.

    PubMed

    Catalanotti, Claudia; Dubini, Alexandra; Subramanian, Venkataramanan; Yang, Wenqiang; Magneschi, Leonardo; Mus, Florence; Seibert, Michael; Posewitz, Matthew C; Grossman, Arthur R

    2012-02-01

    Chlamydomonas reinhardtii, a unicellular green alga, often experiences hypoxic/anoxic soil conditions that activate fermentation metabolism. We isolated three Chlamydomonas mutants disrupted for the pyruvate formate lyase (PFL1) gene; the encoded PFL1 protein catalyzes a major fermentative pathway in wild-type Chlamydomonas cells. When the pfl1 mutants were subjected to dark fermentative conditions, they displayed an increased flux of pyruvate to lactate, elevated pyruvate decarboxylation, ethanol accumulation, diminished pyruvate oxidation by pyruvate ferredoxin oxidoreductase, and lowered H(2) production. The pfl1-1 mutant also accumulated high intracellular levels of lactate, succinate, alanine, malate, and fumarate. 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 it also exhibited a significant increase in the levels of extracellular glycerol, acetate, and intracellular reduced sugars and a decrease in dark, fermentative H(2) production. Whereas wild-type Chlamydomonas fermentation primarily produces formate and ethanol, the double mutant reroutes glycolytic carbon to lactate and glycerol. Although the metabolic adjustments observed in the mutants facilitate NADH reoxidation and sustained glycolysis under dark, anoxic conditions, the observed changes could not have been predicted given our current knowledge of the regulation of fermentation metabolism.

  19. Altered Fermentative Metabolism in Chlamydomonas reinhardtii Mutants Lacking Pyruvate Formate Lyase and Both Pyruvate Formate Lyase and Alcohol Dehydrogenase

    SciTech Connect

    Catalanotti, C.; Dubini, A.; Subramanian, V.; Yang, W. Q.; Magneschi, L.; Mus, F.; Seibert, M.; Posewitz, M. C.; Grossman, A. R.

    2012-02-01

    Chlamydomonas reinhardtii, a unicellular green alga, often experiences hypoxic/anoxic soil conditions that activate fermentation metabolism. We isolated three Chlamydomonas mutants disrupted for the pyruvate formate lyase (PFL1) gene; the encoded PFL1 protein catalyzes a major fermentative pathway in wild-type Chlamydomonas cells. When the pfl1 mutants were subjected to dark fermentative conditions, they displayed an increased flux of pyruvate to lactate, elevated pyruvate decarboxylation, ethanol accumulation, diminished pyruvate oxidation by pyruvate ferredoxin oxidoreductase, and lowered H2 production. The pfl1-1 mutant also accumulated high intracellular levels of lactate, succinate, alanine, malate, and fumarate. 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 it also exhibited a significant increase in the levels of extracellular glycerol, acetate, and intracellular reduced sugars and a decrease in dark, fermentative H2 production. Whereas wild-type Chlamydomonas fermentation primarily produces formate and ethanol, the double mutant reroutes glycolytic carbon to lactate and glycerol. Although the metabolic adjustments observed in the mutants facilitate NADH reoxidation and sustained glycolysis under dark, anoxic conditions, the observed changes could not have been predicted given our current knowledge of the regulation of fermentation metabolism.

  20. Temperature-sensitive rubisco mutant of Chlamydomonas. [Chlamydomonas reinhardtii

    SciTech Connect

    Chen, Z.; Spreitzer, R.J.; Chastain, C.J.

    1987-04-01

    The Chlamydomonas reinhardtii mutant 68-4PP is a temperature-sensitive mutant that lacks photosynthetic ability at 35/sup 0/C, but is able to grow photosynthetically at 25/sup 0/C. Genetic analysis indicated that 68-4PP is a chloroplast mutant that is allelic with known Rubisco large-subunit structural-gene mutants, implying that 68-4PP also resulted from a mutation in the large-subunit gene. The 68-4PP mutant has about 35% of the wild-type level of Rubisco holoenzyme and carboxylase activity when grown at 25/sup 0/C, but it has less than 10% of normal holoenzyme and carboxylase activity when grown at 35/sup 0/C. However, (/sup 35/S)-sulfate pulse labeling showed that Rubisco subunits were synthesized at normal rates at both temperatures. More significantly, the ratio of carboxylase activity in the absence and presence of oxygen at a limiting CO/sub 2/ concentration (6.6 ..mu..M) was about 2.2 for the mutant enzyme, as compared to about 3.0 for the wild-type enzyme. The decreased ratio of the mutant enzyme is maternally inherited, indicating that this reduced oxygen sensitivity results from a mutation in chloroplast DNA. The authors have recently cloned the 68-4PP Rubisco large-subunit gene, and DNA sequencing is in progress.

  1. Efficient phototrophic production of a high-value sesquiterpenoid from the eukaryotic microalga Chlamydomonas reinhardtii.

    PubMed

    Lauersen, Kyle J; Baier, Thomas; Wichmann, Julian; Wördenweber, Robin; Mussgnug, Jan H; Hübner, Wolfgang; Huser, Thomas; Kruse, Olaf

    2016-11-01

    The heterologous expression of terpene synthases in microbial hosts has opened numerous possibilities for bioproduction of desirable metabolites. Photosynthetic microbial hosts present a sustainable alternative to traditional fermentative systems, using freely available (sun)light and carbon dioxide as inputs for bio-production. Here, we report the expression of a patchoulol synthase from Pogostemon cablin Benth in the model green microalga Chlamydomonas reinhardtii. The sesquiterpenoid patchoulol was produced from the alga and was used as a marker of sesquiterpenoid production capacity. A novel strategy for gene loading was employed and patchoulol was produced up to 922±242µgg(-1) CDW in six days. We additionally investigated the effect of carbon source on sesquiterpenoid productivity from C. reinhardtii in scale-up batch cultivations. It was determined that up to 1.03mgL(-1) sesquiterpenoid products could be produced in completely photoautotrophic conditions and that the alga exhibited altered sesquiterpenoid production metabolism related to carbon source.

  2. Diversification of the Core RNA Interference Machinery in Chlamydomonas reinhardtii and the Role of DCL1 in Transposon Silencing

    PubMed Central

    Casas-Mollano, J. Armando; Rohr, Jennifer; Kim, Eun-Jeong; Balassa, Eniko; van Dijk, Karin; Cerutti, Heriberto

    2008-01-01

    Small RNA-guided gene silencing is an evolutionarily conserved process that operates by a variety of molecular mechanisms. In multicellular eukaryotes, the core components of RNA-mediated silencing have significantly expanded and diversified, resulting in partly distinct pathways for the epigenetic control of gene expression and genomic parasites. In contrast, many unicellular organisms with small nuclear genomes seem to have lost entirely the RNA-silencing machinery or have retained only a basic set of components. We report here that Chlamydomonas reinhardtii, a unicellular eukaryote with a relatively large nuclear genome, has undergone extensive duplication of Dicer and Argonaute polypeptides after the divergence of the green algae and land plant lineages. Chlamydomonas encodes three Dicers and three Argonautes with DICER-LIKE1 (DCL1) and ARGONAUTE1 being more divergent than the other paralogs. Interestingly, DCL1 is uniquely involved in the post-transcriptional silencing of retrotransposons such as TOC1. Moreover, on the basis of the subcellular distribution of TOC1 small RNAs and target transcripts, this pathway most likely operates in the nucleus. However, Chlamydomonas also relies on a DCL1-independent, transcriptional silencing mechanism(s) for the maintenance of transposon repression. Our results suggest that multiple, partly redundant epigenetic processes are involved in preventing transposon mobilization in this green alga. PMID:18493041

  3. An improved ARS2-derived nuclear reporter enhances the efficiency and ease of genetic engineering in Chlamydomonas.

    PubMed

    Specht, Elizabeth A; Nour-Eldin, Hussam Hassan; Hoang, Kevin T D; Mayfield, Stephen P

    2015-03-01

    The model alga Chlamydomonas reinhardtii has been used to pioneer genetic engineering techniques for high-value protein and biofuel production from algae. To date, most studies of transgenic Chlamydomonas have utilized the chloroplast genome due to its ease of engineering, with a sizeable suite of reporters and well-characterized expression constructs. The advanced manipulation of algal nuclear genomes has been hampered by limited strong expression cassettes, and a lack of high-throughput reporters. We have improved upon an endogenous reporter gene - the ARS2 gene encoding an arylsulfatase enzyme - that was first cloned and characterized decades ago but has not been used extensively. The new construct, derived from ARS2 cDNA, expresses significantly higher levels of reporter protein and transforms more efficiently, allowing qualitative and quantitative screening using a rapid, inexpensive 96-well assay. The improved arylsulfatase expression cassette was used to screen a new transgene promoter from the ARG7 gene, and found that the ARG7 promoter can express the ARS2 reporter as strongly as the HSP70-RBCS2 chimeric promoter that currently ranks as the best available promoter, thus adding to the list of useful nuclear promoters. This enhanced arylsulfatase reporter construct improves the efficiency and ease of genetic engineering within the Chlamydomonas nuclear genome, with potential application to other algal strains.

  4. A steering mechanism for phototaxis in Chlamydomonas

    PubMed Central

    Bennett, Rachel R.; Golestanian, Ramin

    2015-01-01

    Chlamydomonas shows both positive and negative phototaxis. It has a single eyespot near its equator, and as the cell rotates during the forward motion, the light signal received by the eyespot varies. We use a simple mechanical model of Chlamydomonas that couples the flagellar beat pattern to the light intensity at the eyespot to demonstrate a mechanism for phototactic steering that is consistent with observations. The direction of phototaxis is controlled by a parameter in our model, and the steering mechanism is robust to noise. Our model shows switching between directed phototaxis when the light is on and run-and-tumble behaviour in the dark. PMID:25589576

  5. Stable isotope fractionation in photosynthesis: Analysis of autotrophic competence following transformation of the chloroplast genome of Chlamydomonas

    SciTech Connect

    Boynton, J.E.; Gillham, N.W.; Osmond, C.B.

    1991-06-15

    Isotopic techniques needed to assess the interactions between photosynthesis and respiration in Chlamydomonas have been devised for {sup 13}C, using plate and liquid cultures. The effectiveness of various transformation strategies for the chloroplast psbA gene has been evaluated with respect to their utility in constructing and characterizing strains homoplasmic for site-directed mutations in an otherwise isogenic background. Our analysis of the first site-directed change in the D-1 protein of Chlamydomonas indicates that a second site mutation (arg{sub 238} > lys) in the loop between transmembrane helices IV -- V can partially compensate for the reduced photosynthetic performance that accompanies the atrazine resistant mutation (ser{sub 264} > ala/gly) in this alga and in higher plants grown under high light intensities. 31 refs., 2 figs.

  6. CO2 Biofixation by the Cyanobacterium Spirulina sp. LEB 18 and the Green Alga Chlorella fusca LEB 111 Grown Using Gas Effluents and Solid Residues of Thermoelectric Origin.

    PubMed

    da Silva Vaz, Bruna; Costa, Jorge Alberto Vieira; de Morais, Michele Greque

    2016-01-01

    The concentration of carbon dioxide (CO2) in the atmosphere has increased from 280 to 400 ppm in the last 10 years, and the coal-fired power plants are responsible for approximately 22 % of these emissions. The burning of fossil fuel also produces a great amount of solid waste that causes serious industrial and environmental problems. The biological processes become interesting alternative in combating pollution and developing new products. The objective of this study was to evaluate the CO2 biofixation potential of microalgae that were grown using gaseous effluents and solid residues of thermoelectric origin. The microalgae Chlorella fusca LEB 111 presented higher rate of CO2 biofixation (42.8 %) (p < 0.01) than did Spirulina sp. LEB 18. The values for the CO2 biofixation rates and the kinetic parameters of Spirulina and Chlorella cells grown using combustion gas did not differ significantly from those of cells grown using CO2 and a carbon source in the culture media. These microalgae could be grown using ash derived from coal combustion, using the minerals present in this residue as the source of the essential metals required for their growth and the CO2 derived from the combustion gas as their carbon source.

  7. Analysis of Flagellar Phosphoproteins from Chlamydomonas reinhardtii▿ †

    PubMed Central

    Boesger, Jens; Wagner, Volker; Weisheit, Wolfram; Mittag, Maria

    2009-01-01

    Cilia and flagella are cell organelles that are highly conserved throughout evolution. For many years, the green biflagellate alga Chlamydomonas reinhardtii has served as a model for examination of the structure and function of its flagella, which are similar to certain mammalian cilia. Proteome analysis revealed the presence of several kinases and protein phosphatases in these organelles. Reversible protein phosphorylation can control ciliary beating, motility, signaling, length, and assembly. Despite the importance of this posttranslational modification, the identities of many ciliary phosphoproteins and knowledge about their in vivo phosphorylation sites are still missing. Here we used immobilized metal affinity chromatography to enrich phosphopeptides from purified flagella and analyzed them by mass spectrometry. One hundred forty-one phosphorylated peptides were identified, belonging to 32 flagellar proteins. Thereby, 126 in vivo phosphorylation sites were determined. The flagellar phosphoproteome includes different structural and motor proteins, kinases, proteins with protein interaction domains, and many proteins whose functions are still unknown. In several cases, a dynamic phosphorylation pattern and clustering of phosphorylation sites were found, indicating a complex physiological status and specific control by reversible protein phosphorylation in the flagellum. PMID:19429781

  8. Nitric oxide controls nitrate and ammonium assimilation in Chlamydomonas reinhardtii.

    PubMed

    Sanz-Luque, Emanuel; Ocaña-Calahorro, Francisco; Llamas, Angel; Galvan, Aurora; Fernandez, Emilio

    2013-08-01

    Nitrate and ammonium are major inorganic nitrogen sources for plants and algae. These compounds are assimilated by means of finely regulated processes at transcriptional and post-translational levels. In Chlamydomonas, the expression of several genes involved in high-affinity ammonium (AMT1.1, AMT1.2) and nitrate transport (NRT2.1) as well as nitrate reduction (NIA1) are downregulated by ammonium through a nitric oxide (NO)-dependent mechanism. At the post-translational level, nitrate/nitrite uptake and nitrate reductase (NR) are also inhibited by ammonium, but the mechanisms implicated in this regulation are scarcely known. In this work, the effect of NO on nitrate assimilation and the high-affinity ammonium uptake was addressed. NO inhibited the high-affinity uptake of ammonium and nitrate/nitrite, as well as the NR activity, in a reversible form. In contrast, nitrite reductase and glutamine synthetase activities were not affected. The in vivo and in vitro studies suggested that NR enzyme is inhibited by NO in a mediated process that requires the cell integrity. These data highlight a role of NO in inorganic nitrogen assimilation and suggest that this signalling molecule is an important regulator for the first steps of the pathway.

  9. Singlet oxygen production in Chlamydomonas reinhardtii under heat stress

    PubMed Central

    Prasad, Ankush; Ferretti, Ursula; Sedlářová, Michaela; Pospíšil, Pavel

    2016-01-01

    In the current study, singlet oxygen formation by lipid peroxidation induced by heat stress (40 °C) was studied in vivo in unicellular green alga Chlamydomonas reinhardtii. Primary and secondary oxidation products of lipid peroxidation, hydroperoxide and malondialdehyde, were generated under heat stress as detected using swallow-tailed perylene derivative fluorescence monitored by confocal laser scanning microscopy and high performance liquid chromatography, respectively. Lipid peroxidation was initiated by enzymatic reaction as inhibition of lipoxygenase by catechol and caffeic acid prevented hydroperoxide formation. Ultra-weak photon emission showed formation of electronically excited species such as triplet excited carbonyl, which, upon transfer of excitation energy, leads to the formation of either singlet excited chlorophyll or singlet oxygen. Alternatively, singlet oxygen is formed by direct decomposition of hydroperoxide via Russell mechanisms. Formation of singlet oxygen was evidenced by the nitroxyl radical 2,2,6,6-tetramethylpiperidine-1-oxyl detected by electron paramagnetic resonance spin-trapping spectroscopy and the imaging of green fluorescence of singlet oxygen sensor green detected by confocal laser scanning microscopy. Suppression of singlet oxygen formation by lipoxygenase inhibitors indicates that singlet oxygen may be formed via enzymatic lipid peroxidation initiated by lipoxygenase. PMID:26831215

  10. Gene Expression Profiling of Flagellar Disassembly in Chlamydomonas reinhardtii

    PubMed Central

    Chamberlain, Kara L.; Miller, Steven H.; Keller, Laura R.

    2008-01-01

    Flagella are sensory organelles that interact with the environment through signal transduction and gene expression networks. We used microarray profiling to examine gene regulation associated with flagellar length change in the green alga Chlamydomonas reinhardtii. Microarrays were probed with fluorescently labeled cDNAs synthesized from RNA extracted from cells before and during flagellar assembly or disassembly. Evaluation of the gene expression profiles identified >100 clones showing at least a twofold change in expression during flagellar length changes. Products of these genes are associated not only with flagellar structure and motility but also with other cellular responses, including signal transduction and metabolism. Expression of specific genes from each category was further characterized at higher resolution by using quantitative real-time PCR (qRT–PCR). Analysis and comparison of the gene expression profiles coupled to flagellar assembly and disassembly revealed that each process involves a new and uncharacterized whole-cell response to flagellar length changes. This analysis lays the groundwork for a more comprehensive understanding of the cellular and molecular networks regulating flagellar length changes. PMID:18493036

  11. Photosynthetic characteristics of a multicellular green alga Volvox carteri in response to external CO2 levels possibly regulated by CCM1/CIA5 ortholog.

    PubMed

    Yamano, Takashi; Fujita, Akimitsu; Fukuzawa, Hideya

    2011-09-01

    When CO(2) supply is limited, aquatic photosynthetic organisms induce a CO(2)-concentrating mechanism (CCM) and acclimate to the CO(2)-limiting environment. Although the CCM is well studied in unicellular green algae such as Chlamydomonas reinhardtii, physiological aspects of the CCM and its associated genes in multicellular algae are poorly understood. In this study, by measuring photosynthetic affinity for CO(2), we present physiological data in support of a CCM in a multicellular green alga, Volvox carteri. The low-CO(2)-grown Volvox cells showed much higher affinity for inorganic carbon compared with high-CO(2)-grown cells. Addition of ethoxyzolamide, a membrane-permeable carbonic anhydrase inhibitor, to the culture remarkably reduced the photosynthetic affinity of low-CO(2) grown Volvox cells, indicating that an intracellular carbonic anhydrase contributed to the Volvox CCM. We also isolated a gene encoding a protein orthologous to CCM1/CIA5, a master regulator of the CCM in Chlamydomonas, from Volvox carteri. Volvox CCM1 encoded a protein with 701 amino acid residues showing 51.1% sequence identity with Chlamydomonas CCM1. Comparison of Volvox and Chlamydomonas CCM1 revealed a highly conserved N-terminal region containing zinc-binding amino acid residues, putative nuclear localization and export signals, and a C-terminal region containing a putative LXXLL protein-protein interaction motif. Based on these results, we discuss the physiological and genetic aspects of the CCM in Chlamydomonas and Volvox.

  12. Expression of bkt and bch genes from Haematococcus pluvialis in transgenic Chlamydomonas.

    PubMed

    Zheng, KaiJing; Wang, ChaoGang; Xiao, Ming; Chen, Jun; Li, JianCheng; Hu, ZhangLi

    2014-10-01

    β-carotene ketolase and β-carotene hydroxylase encoded by bkt and bch, respectively, are key enzymes required for astaxanthin biosynthesis in Haematococcu pluvialis 34-1n. Two expression vectors containing cDNA sequences of bkt and bch were constructed and co-transformed into cell-wall-deficient Chlamydomonas reinhardtii CC-849. Transgenic algae were screened on TAP agar plates containing 10 μg mL(-1) Zeomycin. PCR-Southern analysis showed that bkt and bch were integrated into the genomes of C. reinhardtii. Transcripts of bkt and bch were further confirmed by RT-PCR-Southern analysis. Compared with the wild type, transgenic algae produced 29.04% and 30.27% more carotenoids and xanthophylls, respectively. Moreover, the transgenic algae could accumulate 34% more astaxanthin than wild type. These results indicate that foreign bkt and bch genes were successfully translated into β-carotene ketolase and β-carotene hydroxylase, which were responsible for catalyzing the biosynthesis of astaxanthin in transgenic algae.

  13. Harvesting fresh water and marine algae by magnetic separation: screening of separation parameters and high gradient magnetic filtration.

    PubMed

    Cerff, Martin; Morweiser, Michael; Dillschneider, Robert; Michel, Aymeé; Menzel, Katharina; Posten, Clemens

    2012-08-01

    In this study, the focus is on magnetic separation of fresh water algae Chlamydomonas reinhardtii and Chlorella vulgaris as well as marine algae Phaeodactylum tricornutum and Nannochloropsis salina by means of silica-coated magnetic particles. Due to their small size and low biomass concentrations, harvesting algae by conventional methods is often inefficient and cost-consuming. Magnetic separation is a powerful tool to capture algae by adsorption to submicron-sized magnetic particles. Hereby, separation efficiency depends on parameters such as particle concentration, pH and medium composition. Separation efficiencies of >95% were obtained for all algae while maximum particle loads of 30 and 77 g/g were measured for C. reinhardtii and P. tricornutum at pH 8 and 12, respectively. This study highlights the potential of silica-coated magnetic particles for the removal of fresh water and marine algae by high gradient magnetic filtration and provides critical discussion on future improvements.

  14. Bioaccumulation of nickel by algae

    SciTech Connect

    Wang, H.K.; Wood, J.M.

    1984-02-01

    Six strains of algae and one Euglena sp. were tested for their ability to bioaccumulate nickel. Radioactive /sup 63/Ni was used together with a microplate technique to determine the conditions for nickel removal by axenic cultures of cyanobacteria, green algae, and one euglenoid. The cyanobacteria tested were found to be more sensitive to nickel toxicity than the green algae or the Euglena sp. The concentration factor (CF) for nickel was determined under a variety of conditions and found to be in the range from 0 to 3.0 x 10/sup 3/. The effect of environmental variables on nickel uptake was examined, and a striking pH effect for biaccumulation was observed, with most of the algal strains accumulating nickel optimally at approximately pH 8.0. Competition experiments for binding sites between nickel and other cations as well as with other complexing anions, showed that /sup 63/Ni uptake was affected only by cobalt and by humic acids.

  15. Expression of human antibodies in eukaryotic micro-algae.

    PubMed

    Mayfield, Stephen P; Franklin, Scott E

    2005-03-07

    Protein based therapeutics have enjoyed great success over the past decade. Unfortunately, with this clinical success comes a heavy price tag, owing to the inherently high costs of capitalization and production using mammalian cell fermentation. To address this problem, we have begun developing a system for the expression of recombinant proteins in the unicellular eukaryotic green algae, Chlamydomonas reinhardtii, leading to the production of human IgA single chain antibodies. The expression of human monoclonal antibodies in C. reinhardtii offers an attractive alternative to traditional mammalian based expression systems for several reasons, including an ability to rapidly obtain stable plastid and nuclear transformants, coupled with inherently low costs of capitalization and production.

  16. CrGNAT gene regulates excess copper accumulation and tolerance in Chlamydomonas reinhardtii.

    PubMed

    Wang, Ye; Cheng, Zhen Zhen; Chen, Xi; Zheng, Qi; Yang, Zhi Min

    2015-11-01

    Excess copper (Cu) in environment affects the growth and metabolism of plants and green algae. However, the molecular mechanism for regulating plant tolerance to excess Cu is not fully understood. Here, we report a gene CrGNAT enconding an acetyltransferase in Chlamydomonas reinhardtii and identified its role in regulating tolerance to Cu toxicity. Expression of CrGNAT was significantly induced by 75-400μM Cu. The top induction occurred at 100μM. Transgenic algae overexpressing CrGNAT (35S::CrGNAT) in C. reinhardtii showed high tolerance to excess Cu, with improved cell population, chlorophyll accumulation and photosynthesis efficiency, but with low degree of oxidation with regard to reduced hydrogen peroxide, lipid peroxides and non-protein thiol compounds. In contrast, CrGNAT knock-down lines with antisense led to sensitivity to Cu stress. 35S::CrGNAT algae accumulated more Cu and other metals (Zn, Fe, Cu, Mn and Mg) than wild-type, whereas the CrGNAT down-regulated algae (35S::AntiCrGNAT) had moderate levels of Cu and Mn, but no effects on Zn, Fe and Mg accumulation as compared to wild-type. The elevated metal absorption in CrGNAT overexpression algae implies that the metals can be removed from water media. Quantitative RT-PCR analysis revealed that expression of two genes encoding N-lysine histone methyltransferases was repressed in 35S::CrGNAT algae, suggesting that CrGNAT-regulated algal tolerance to Cu toxicity is likely associated with histone methylation and chromatin remodeling. The present work provided an example a basis to develop techniques for environmental restoration of metal-contaminated aquatic ecosystems.

  17. A comparison of hydrogen photoproduction by sulfur-deprived Chlamydomonas reinhardtii under different growth conditions.

    PubMed

    Kosourov, Sergey; Patrusheva, Elena; Ghirardi, Maria L; Seibert, Michael; Tsygankov, Anatoly

    2007-03-10

    Continuous photoproduction of H(2) by the green alga, Chlamydomonas reinhardtii, is observed after incubating the cultures for about a day in the absence of sulfate and in the presence of acetate. Sulfur deprivation causes the partial and reversible inactivation of photosynthetic O(2) evolution in algae, resulting in the light-induced establishment of anaerobic conditions in sealed photobioreactors, expression of two [FeFe]-hydrogenases in the cells, and H(2) photoproduction for several days. We have previously demonstrated that sulfur-deprived algal cultures can produce H(2) gas in the absence of acetate, when appropriate experimental protocols were used (Tsygankov, A.A., Kosourov, S.N., Tolstygina, I.V., Ghirardi, M.L., Seibert, M., 2006. Hydrogen production by sulfur-deprived Chlamydomonas reinhardtii under photoautotrophic conditions. Int. J. Hydrogen Energy 31, 1574-1584). We now report the use of an automated photobioreactor system to compare the effects of photoautotrophic, photoheterotrophic and photomixotrophic growth conditions on the kinetic parameters associated with the adaptation of the algal cells to sulfur deprivation and H(2) photoproduction. This was done under the experimental conditions outlined in the above reference, including controlled pH. From this comparison we show that both acetate and CO(2) are required for the most rapid inactivation of photosystem II and the highest yield of H(2) gas production. Although, the presence of acetate in the system is not critical for the process, H(2) photoproduction under photoautotrophic conditions can be increased by optimizing the conditions for high starch accumulation. These results suggest ways of engineering algae to improve H(2) production, which in turn may have a positive impact on the economics of applied systems for H(2) production.

  18. Marine Bacteria from Danish Coastal Waters Show Antifouling Activity against the Marine Fouling Bacterium Pseudoalteromonas sp. Strain S91 and Zoospores of the Green Alga Ulva australis Independent of Bacteriocidal Activity▿†

    PubMed Central

    Bernbom, Nete; Ng, Yoke Yin; Kjelleberg, Staffan; Harder, Tilmann; Gram, Lone

    2011-01-01

    The aims of this study were to determine if marine bacteria from Danish coastal waters produce antifouling compounds and if antifouling bacteria could be ascribed to specific niches or seasons. We further assess if antibacterial effect is a good proxy for antifouling activity. We isolated 110 bacteria with anti-Vibrio activity from different sample types and locations during a 1-year sampling from Danish coastal waters. The strains were identified as Pseudoalteromonas, Phaeobacter, and Vibrionaceae based on phenotypic tests and partial 16S rRNA gene sequence similarity. The numbers of bioactive bacteria were significantly higher in warmer than in colder months. While some species were isolated at all sampling locations, others were niche specific. We repeatedly isolated Phaeobacter gallaeciensis at surfaces from one site and Pseudoalteromonas tunicata at two others. Twenty-two strains, representing the major taxonomic groups, different seasons, and isolation strategies, were tested for antiadhesive effect against the marine biofilm-forming bacterium Pseudoalteromonas sp. strain S91 and zoospores of the green alga Ulva australis. The antiadhesive effects were assessed by quantifying the number of strain S91 or Ulva spores attaching to a preformed biofilm of each of the 22 strains. The strongest antifouling activity was found in Pseudoalteromonas strains. Biofilms of Pseudoalteromonas piscicida, Pseudoalteromonas tunicata, and Pseudoalteromonas ulvae prevented Pseudoalteromonas S91 from attaching to steel surfaces. P. piscicida killed S91 bacteria in the suspension cultures, whereas P. tunicata and P. ulvae did not; however, they did prevent adhesion by nonbactericidal mechanism(s). Seven Pseudoalteromonas species, including P. piscicida and P. tunicata, reduced the number of settling Ulva zoospores to less than 10% of the number settling on control surfaces. The antifouling alpP gene was detected only in P. tunicata strains (with purple and yellow pigmentation), so

  19. Measurement of ethanol formation in single living cells of Chlamydomonas reinhardtii using synchrotron Fourier Transform Infrared spectromicroscopy

    SciTech Connect

    Goff, Kira L.; Quaroni, Luca; Pedersen, Tor; Wilson, Kenneth E.

    2010-02-03

    We demonstrate the capability of Fourier-Transform Infra-Red (FITR) spectroscopy to detect metabolite formation by the unicellular algae Chlamydomonas reinhardtii in solution. We show that using a synchrotron source in the microscopy configuration provides a sufficient s/n ratio to detect small molecular species accumulating at a single cell, allowing an increased sensitivity relative to measurements of bulk cultures. The formation of small molecular species, including ethanol and at least one carbonyl containing compound, can be detected with a time resolution of the order of one minute.

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

    SciTech Connect

    Not Available

    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.

  1. Evidence for a photoprotective function for secondary carotenoids of snow algae

    SciTech Connect

    Bidigare, R.R.; Ondrusek, M.E. ); Kennicutt, M.C. II ); Iturriaga, R. ); Harvey, H.R. ); Hoham, R.W. ); Macko, S.A. )

    1993-08-01

    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.

  2. Rapid construction and screening of artificial microRNA systems in Chlamydomonas reinhardtii.

    PubMed

    Hu, Jinlu; Deng, Xuan; Shao, Ning; Wang, Gaohong; Huang, Kaiyao

    2014-09-01

    The unicellular green algae Chlamydomonas reinhardtii is a classic model for the study of flagella/cilia and photosynthesis, and it has recently been exploited for producing biopharmaceuticals and biofuel. Due to the low frequency of homologous recombination, reverse genetic manipulation in Chlamydomonas relies mainly on miRNA- and siRNA-based knockdown methods. However, the difficulty in constructing artificial miRNA vectors, laborious screening of knockdown transformants, and undesired epigenetic silencing of exogenous miRNA constructs limit their application. We have established a one-step procedure to construct an artificial miRNA precursor by annealing eight oligonucleotides of approximately 40 nucleotides. In the final construct, the Gaussia princeps luciferase gene (G-Luc) is positioned between the promoter and the artificial miRNA precursor so that knockdown strains may quickly be screened by visualizing luciferase luminescence using a photon-counting camera. Furthermore, the luciferase activity of transformants correlates with the knockdown level of two test target proteins: the chloroplast protein VIPP1 (vesicle inducing protein in plastids 1) and the flagellar protein CDPK3 (calcium-dependent protein kinase 3). Adding an intron from RBCS2 (ribulose bisphosphate carboxylase/oxygenase small subunit 2) to the miRNA construct enhanced both the luciferase activity and the miRNA knockdown efficiency. A second miRNA vector incorporated the promoter of the nitrate reductase gene to allow inducible expression of the artificial miRNA. These vectors will facilitate application of the artificial miRNA and provide tools for studying the mechanism of epigenetics in Chlamydomonas, and may also be adapted for use in other model organisms.

  3. Evaluation of filamentous green algae as feedstocks for biofuel production.

    PubMed

    Zhang, Wei; Zhao, Yonggang; Cui, Binjie; Wang, Hui; Liu, Tianzhong

    2016-11-01

    Compared with unicellular microalgae, filamentous algae have high resistance to grazer-predation and low-cost recovery in large-scale production. Green algae, as the most diverse group of algae, included numerous filamentous genera and species. In this study, records of filamentous genera and species in green algae were firstly censused and classified. Then, seven filamentous strains subordinated in different genera were cultivated in bubbled-column to investigate their growth rate and energy molecular (lipid and starch) capacity. Four strains including Stigeoclonium sp., Oedogonium nodulosum, Hormidium sp. and Zygnema extenue were screened out due to their robust growth. And they all could accumulate triacylglycerols and starch in their biomass, but with different capacity. After nitrogen starvation, Hormidium sp. and Oedogonium nodulosum respectively exhibited high capacity of lipid (45.38% in dry weight) and starch (46.19% in dry weight) accumulation, which could be of high potential as feedstocks for biodiesel and bioethanol production.

  4. Green Algae as Model Organisms for Biological Fluid Dynamics.

    PubMed

    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 millimetres), 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.

  5. Green Algae as Model Organisms for Biological Fluid Dynamics

    NASA Astrophysics Data System (ADS)

    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.

  6. Green Algae as Model Organisms for Biological Fluid Dynamics*

    PubMed Central

    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 millimetres), 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. PMID:26594068

  7. Characterization of a Mutant Deficient for Ammonium and Nitric Oxide Signalling in the Model System Chlamydomonas reinhardtii

    PubMed Central

    Sanz-Luque, Emanuel; Ocaña-Calahorro, Francisco; Galván, Aurora; Fernández, Emilio; de Montaigu, Amaury

    2016-01-01

    The ubiquitous signalling molecule Nitric Oxide (NO) is characterized not only by the variety of organisms in which it has been described, but also by the wealth of biological processes that it regulates. In contrast to the expanding repertoire of functions assigned to NO, however, the mechanisms of NO action usually remain unresolved, and genes that work within NO signalling cascades are seldom identified. A recent addition to the list of known NO functions is the regulation of the nitrogen assimilation pathway in the unicellular alga Chlamydomonas reinhardtii, a well-established model organism for genetic and molecular studies that offers new possibilities in the search for mediators of NO signalling. By further exploiting a collection of Chlamydomonas insertional mutant strains originally isolated for their insensitivity to the ammonium (NH4+) nitrogen source, we found a mutant which, in addition to its ammonium insensitive (AI) phenotype, was not capable of correctly sensing the NO signal. Similarly to what had previously been described in the AI strain cyg56, the expression of nitrogen assimilation genes in the mutant did not properly respond to treatments with various NO donors. Complementation experiments showed that NON1 (NO Nitrate 1), a gene that encodes a protein containing no known functional domain, was the gene underlying the mutant phenotype. Beyond the identification of NON1, our findings broadly demonstrate the potential for Chlamydomonas reinhardtii to be used as a model system in the search for novel components of gene networks that mediate physiological responses to NO. PMID:27149516

  8. Magnetic separation of algae

    SciTech Connect

    Nath, Pulak; Twary, Scott N.

    2016-04-26

    Described herein are methods and systems for harvesting, collecting, separating and/or dewatering algae using iron based salts combined with a magnetic field gradient to separate algae from an aqueous solution.

  9. HISN3 mediates adaptive response of Chlamydomonas reinhardtii to excess nickel.

    PubMed

    Zheng, Qi; Cheng, Zhen Zhen; Yang, Zhi Min

    2013-12-01

    Investigation of genes for heavy metal [e.g. nickel (Ni) and zinc (Zn)] absorption and detoxification in green algae is of great importance because some of the metals have become one of the major contaminants in the aquatic ecosystem. In plants, overload of heavy metals modifies many aspects of biological processes. However, the mechanisms by which heavy metals exert detrimental effects are not fully understood. The present study identified a biological role for HISN3 (the gene coding for phosphoribosylformimino-5-aminoimidazole carboxamide ribonucleotide isomerase) in regulating the response of Chlamydomonas reinhardtii, a unicellular green alga, to Ni toxicity. In higher plants, HISN3 encodes an enzyme catalyzing the fourth step in the histidine biosynthesis pathway, but its functional importance is yet to be identified. Transgenic algae overexpressing HISN3 in C. reinhardtii showed high tolerance to excess Ni, with a 48.3-57.4% increase in cell population and moderate histidine accumulation compared with the wild type. HISN3 overexpression improved accumulation of Chl and photosynthesis efficiency, but suppressed Ni-induced generation of reactive oxygen species and lipid peroxides. Interestingly, more Ni and other metals [Zn, iron (Fe), copper (Cu), manganese (Mn) and magnesium (Mg)] were accumulated in HISN3-overexpressing cells than in the wild type. In contrast, RNA interference of HISN3 depressed Ni accumulation but caused cellular sensitivity to Ni. The elevated metal absorption in the HISN3-overexpressing algae implies that the metals can be removed from water media. Thus, our work presents an example for algae genetically designed to improve tolerance to metal toxicity and environmental restoration of metal-contaminated aquatic ecosystems.

  10. Culture of microalgae Chlamydomonas reinhardtii in wastewater for biomass feedstock production.

    PubMed

    Kong, Qing-xue; Li, Ling; Martinez, Blanca; Chen, Paul; Ruan, Roger

    2010-01-01

    The objective of this research was to develop large-scale technologies to produce oil-rich algal biomass from wastewater. The experiments were conducted using Erlenmeyer flasks and biocoil photobioreactor. Chlamydomonas reinhardtii was grown in artificial media and wastewaters taken from three different stages of the treatment process, namely, influent, effluent, and centrate. Each of wastewaters contained different levels of nutrients. The specific growth rate of C. reinhardtii in different cultures was monitored over a period of 10 days. The biomass yield of microalgae and associated nitrogen and phosphorous removal were evaluated. Effects of CO(2) and pH on the growth were also studied. The level of nutrients greatly influenced algae growth. High levels of nutrients seem to inhibit algae growth in the beginning, but provided sustained growth to a high degree. The studies have shown that the optimal pH for C. reinhardtii is in the range of 7.5. An injection of air and a moderate amount of CO(2) promoted algae growth. However, too much CO(2) inhibited algae growth due to a significant decrease in pH. The experimental results showed that algal dry biomass yield reached a maximum of 2.0 g L(-1) day(-1) in the biocoil. The oil content of microalgae of C. reinhardtii was 25.25% (w/w) in dry biomass weight. In the biocoil, 55.8 mg nitrogen and 17.4 mg phosphorus per liter per day were effectively removed from the centrate wastewater. Ferric chloride was found to be an effective flocculent that helps the algae settle for easy harvest and separation from the culture media.

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

    PubMed

    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.

  12. MEETING: Chlamydomonas Annotation Jamboree - October 2003

    SciTech Connect

    Grossman, Arthur R

    2007-04-13

    Shotgun sequencing of the nuclear genome of Chlamydomonas reinhardtii (Chlamydomonas throughout) was performed at an approximate 10X coverage by JGI. Roughly half of the genome is now contained on 26 scaffolds, all of which are at least 1.6 Mb, and the coverage of the genome is ~95%. There are now over 200,000 cDNA sequence reads that we have generated as part of the Chlamydomonas genome project (Grossman, 2003; Shrager et al., 2003; Grossman et al. 2007; Merchant et al., 2007); other sequences have also been generated by the Kasuza sequence group (Asamizu et al., 1999; Asamizu et al., 2000) or individual laboratories that have focused on specific genes. Shrager et al. (2003) placed the reads into distinct contigs (an assemblage of reads with overlapping nucleotide sequences), and contigs that group together as part of the same genes have been designated ACEs (assembly of contigs generated from EST information). All of the reads have also been mapped to the Chlamydomonas nuclear genome and the cDNAs and their corresponding genomic sequences have been reassembled, and the resulting assemblage is called an ACEG (an Assembly of contiguous EST sequences supported by genomic sequence) (Jain et al., 2007). Most of the unique genes or ACEGs are also represented by gene models that have been generated by the Joint Genome Institute (JGI, Walnut Creek, CA). These gene models have been placed onto the DNA scaffolds and are presented as a track on the Chlamydomonas genome browser associated with the genome portal (http://genome.jgi-psf.org/Chlre3/Chlre3.home.html). Ultimately, the meeting grant awarded by DOE has helped enormously in the development of an annotation pipeline (a set of guidelines used in the annotation of genes) and resulted in high quality annotation of over 4,000 genes; the annotators were from both Europe and the USA. Some of the people who led the annotation initiative were Arthur Grossman, Olivier Vallon, and Sabeeha Merchant (with many individual

  13. Taxonomic revision of Chlamydomonas subg. Amphichloris (Volvocales, Chlorophyceae), with resurrection of the genus Dangeardinia and descriptions of Ixipapillifera gen. nov. and Rhysamphichloris gen. nov.

    PubMed

    Nakada, Takashi; Tomita, Masaru; Wu, Jiunn-Tzong; Nozaki, Hisayoshi

    2016-04-01

    Chlamydomonas (Cd.) is one of the largest but most polyphyletic genera of freshwater unicellular green algae. It consists of 400-600 morphological species and requires taxonomic revision. Toward reclassification, each morphologically defined classical subgenus (or subgroup) should be examined using culture strains. Chlamydomonas subg. Amphichloris is characterized by a central nucleus between two axial pyrenoids, however, the phylogenetic structure of this subgenus has yet to be examined using molecular data. Here, we examined 12 strains including six newly isolated strains, morphologically identified as Chlamydomonas subg. Amphichloris, using 18S rRNA gene phylogeny, light microscopy, and mitochondria fluorescent microscopy. Molecular phylogenetic analyses revealed three independent lineages of the subgenus, separated from the type species of Chlamydomonas, Cd. reinhardtii. These three lineages were further distinguished from each other by light and fluorescent microscopy-in particular by the morphology of the papillae, chloroplast surface, stigmata, and mitochondria-and are here assigned to three genera: Dangeardinia emend., Ixipapillifera gen. nov., and Rhysamphichloris gen. nov. Based on the molecular and morphological data, two to three species were recognized in each genus, including one new species, I. pauromitos. In addition, Cd. deasonii, which was previously assigned to subgroup "Pleiochloris," was included in the genus Ixipapillifera as I. deasonii comb. nov.

  14. Altered Fermentative Metabolism in Chlamydomonas reinhardtii Mutants Lacking Pyruvate Formate Lyase and Both Pyruvate Formate Lyase and Alcohol Dehydrogenase[W

    PubMed Central

    Catalanotti, Claudia; Dubini, Alexandra; Subramanian, Venkataramanan; Yang, Wenqiang; Magneschi, Leonardo; Mus, Florence; Seibert, Michael; Posewitz, Matthew C.; Grossman, Arthur R.

    2012-01-01

    Chlamydomonas reinhardtii, a unicellular green alga, often experiences hypoxic/anoxic soil conditions that activate fermentation metabolism. We isolated three Chlamydomonas mutants disrupted for the pyruvate formate lyase (PFL1) gene; the encoded PFL1 protein catalyzes a major fermentative pathway in wild-type Chlamydomonas cells. When the pfl1 mutants were subjected to dark fermentative conditions, they displayed an increased flux of pyruvate to lactate, elevated pyruvate decarboxylation, ethanol accumulation, diminished pyruvate oxidation by pyruvate ferredoxin oxidoreductase, and lowered H2 production. The pfl1-1 mutant also accumulated high intracellular levels of lactate, succinate, alanine, malate, and fumarate. 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 it also exhibited a significant increase in the levels of extracellular glycerol, acetate, and intracellular reduced sugars and a decrease in dark, fermentative H2 production. Whereas wild-type Chlamydomonas fermentation primarily produces formate and ethanol, the double mutant reroutes glycolytic carbon to lactate and glycerol. Although the metabolic adjustments observed in the mutants facilitate NADH reoxidation and sustained glycolysis under dark, anoxic conditions, the observed changes could not have been predicted given our current knowledge of the regulation of fermentation metabolism. PMID:22353371

  15. Photoinduced electric currents in carotenoid-deficient Chlamydomonas mutants reconstituted with retinal and its analogs.

    PubMed Central

    Sineshchekov, O A; Govorunova, E G; Dér, A; Keszthelyi, L; Nultsch, W

    1994-01-01

    Reconstitution of the photoelectric responses involved in photosensory transduction in "blind" cells of Chlamydomonas reinhardtii carotenoid-deficient mutants was studied by means of a recently developed population method. Both the photoreceptor current and the regenerative response can be restored by addition of all-trans-retinal, 9-demethyl-retinal, or dimethyl-octatrienal, while the retinal analogs prevented from 13-cis/trans isomerization, 13-demethyl-retinal and citral, are not effective. Fluence dependence, spectral sensitivity, and effect of hydroxylamine treatment on retinal-induced photoelectric responses are similar to those found earlier in green strains of Chlamydomonas, although an alternative mechanism of antenna directivity in white cells of reconstituted "blind" mutants (likely based on the focusing effect of the transparent cell bodies) leads to the reversed sign of phototaxis in mutant cells under the same conditions. The results obtained indicate that both photoreceptor current and regenerative response are initiated by the same or similar rhodopsins with arhaebacterial-like chromophore(s) and prove directly the earlier suggested identity of the photoreceptor pigment(s) involved in photomotile and photoelectric responses in flagellated algae. PMID:8075341

  16. System response of metabolic networks in Chlamydomonas reinhardtii to total available ammonium.

    PubMed

    Lee, Do Yup; Park, Jeong-Jin; Barupal, Dinesh K; Fiehn, Oliver

    2012-10-01

    Drastic alterations in macronutrients are known to cause large changes in biochemistry and gene expression in the photosynthetic alga Chlamydomonas reinhardtii. However, metabolomic and proteomic responses to subtle reductions in macronutrients have not yet been studied. When ammonium levels were reduced by 25-100% compared with control cultures, ammonium uptake and growth rates were not affected at 25% or 50% nitrogen-reduction for 28 h. However, primary metabolism and enzyme expression showed remarkable changes at acute conditions (4 h and 10 h after ammonium reduction) compared with chronic conditions (18 h and 28 h time points). Responses of 145 identified metabolites were quantified using gas chromatography-time of flight mass spectrometry; 495 proteins (including 187 enzymes) were monitored using liquid chromatography-ion trap mass spectrometry with label-free spectral counting. Stress response and carbon assimilation processes (Calvin cycle, acetate uptake and chlorophyll biosynthesis) were altered first, in addition to increase in enzyme contents for lipid biosynthesis and accumulation of short chain free fatty acids. Nitrogen/carbon balance metabolism was found changed only under chronic conditions, for example in the citric acid cycle and amino acid metabolism. Metabolism in Chlamydomonas readily responds to total available media nitrogen with temporal increases in short-chain free fatty acids and turnover of internal proteins, long before nitrogen resources are depleted.

  17. Characterization of DNA repair deficient strains of Chlamydomonas reinhardtii generated by insertional mutagenesis.

    PubMed

    Plecenikova, Andrea; Slaninova, Miroslava; Riha, Karel

    2014-01-01

    While the mechanisms governing DNA damage response and repair are fundamentally conserved, cross-kingdom comparisons indicate that they differ in many aspects due to differences in life-styles and developmental strategies. In photosynthetic organisms these differences have not been fully explored because gene-discovery approaches are mainly based on homology searches with known DDR/DNA repair proteins. Here we performed a forward genetic screen in the green algae Chlamydomonas reinhardtii to identify genes deficient in DDR/DNA repair. We isolated five insertional mutants that were sensitive to various genotoxic insults and two of them exhibited altered efficiency of transgene integration. To identify genomic regions disrupted in these mutants, we established a novel adaptor-ligation strategy for the efficient recovery of the insertion flanking sites. Four mutants harbored deletions that involved known DNA repair factors, DNA Pol zeta, DNA Pol theta, SAE2/COM1, and two neighbouring genes encoding ERCC1 and RAD17. Deletion in the last mutant spanned two Chlamydomonas-specific genes with unknown function, demonstrating the utility of this approach for discovering novel factors involved in genome maintenance.

  18. Experimental Genome-Wide Determination of RNA Polyadenylation in Chlamydomonas reinhardtii

    PubMed Central

    Bell, Stephen A.; Shen, Chi; Brown, Alishea; Hunt, Arthur G.

    2016-01-01

    The polyadenylation of RNA is a near-universal feature of RNA metabolism in eukaryotes. This process has been studied in the model alga Chlamydomonas reinhardtii using low-throughput (gene-by-gene) and high-throughput (transcriptome sequencing) approaches that recovered poly(A)-containing sequence tags which revealed interesting features of this critical process in Chlamydomonas. In this study, RNA polyadenylation has been studied using the so-called Poly(A) Tag Sequencing (PAT-Seq) approach. Specifically, PAT-Seq was used to study poly(A) site choice in cultures grown in four different media types—Tris-Phosphate (TP), Tris-Phosphate-Acetate (TAP), High-Salt (HS), and High-Salt-Acetate (HAS). The results indicate that: 1. As reported before, the motif UGUAA is the primary, and perhaps sole, cis-element that guides mRNA polyadenylation in the nucleus; 2. The scope of alternative polyadenylation events with the potential to change the coding sequences of mRNAs is limited; 3. Changes in poly(A) site choice in cultures grown in the different media types are very few in number and do not affect protein-coding potential; 4. Organellar polyadenylation is considerable and affects primarily ribosomal RNAs in the chloroplast and mitochondria; and 5. Organellar RNA polyadenylation is a dynamic process that is affected by the different media types used for cell growth. PMID:26730730

  19. A galactoglycerolipid lipase is required for triacylglycerol accumulation and survival following nitrogen deprivation in Chlamydomonas reinhardtii.

    PubMed

    Li, Xiaobo; Moellering, Eric R; Liu, Bensheng; Johnny, Cassandra; Fedewa, Marie; Sears, Barbara B; Kuo, Min-Hao; Benning, Christoph

    2012-11-01

    Following N deprivation, microalgae accumulate triacylglycerols (TAGs). To gain mechanistic insights into this phenomenon, we identified mutants with reduced TAG content following N deprivation in the model alga Chlamydomonas reinhardtii. In one of the mutants, the disruption of a galactoglycerolipid lipase-encoding gene, designated PLASTID GALACTOGLYCEROLIPID DEGRADATION1 (PGD1), was responsible for the primary phenotype: reduced TAG content, altered TAG composition, and reduced galactoglycerolipid turnover. The recombinant PGD1 protein, which was purified from Escherichia coli extracts, hydrolyzed monogalactosyldiacylglycerol into its lyso-lipid derivative. In vivo pulse-chase labeling identified galactoglycerolipid pools as a major source of fatty acids esterified in TAGs following N deprivation. Moreover, the fatty acid flux from plastid lipids to TAG was decreased in the pgd1 mutant. Apparently, de novo-synthesized fatty acids in Chlamydomonas reinhardtii are, at least partially, first incorporated into plastid lipids before they enter TAG synthesis. As a secondary effect, the pgd1 mutant exhibited a loss of viability following N deprivation, which could be avoided by blocking photosynthetic electron transport. Thus, the pgd1 mutant provides evidence for an important biological function of TAG synthesis following N deprivation, namely, relieving a detrimental overreduction of the photosynthetic electron transport chain.

  20. Generation of the heterodimeric precursor GP3 of the Chlamydomonas cell wall.

    PubMed

    Voigt, Jürgen; Kiess, Michael; Getzlaff, Rita; Wöstemeyer, Johannes; Frank, Ronald

    2010-09-01

    The cell wall of the unicellular green alga Chlamydomonas reinhardtii exclusively consists of hydroxyproline-containing glycoproteins. Protein chemical analysis of its polypeptide constituents was hindered by their cross-linking via peroxidase-catalysed intermolecular isodityrosine formation and transaminase-dependent processes. To overcome this problem, we have identified putative soluble precursors using polyclonal antibodies raised against deglycosylation products of the highly purified insoluble wall fraction and analysed their amino acid sequence. The occurrence of the corresponding polypeptide in the insoluble glycoprotein framework was finally probed by epitope mapping of the polyclonal antibodies using overlapping scan peptides which, together, cover the whole amino acid sequence of the putative precursor. As a control, peptide fragments released from the insoluble wall fraction by trypsin treatment were analysed by mass spectroscopy. By this approach, the heterodimeric, chaotrope-soluble glycoprotein GP3 proved to be a constituent of the insoluble extracellular matrix of Chlamydomonas reinhardtii. Furthermore, we have shown that the polypeptide backbones of both GP3 subunits are encoded by the same gene and differ by a C-terminal truncation in the case of GP3A.

  1. CRISPR/Cas9-induced knockout and knock-in mutations in Chlamydomonas reinhardtii.

    PubMed

    Shin, Sung-Eun; Lim, Jong-Min; Koh, Hyun Gi; Kim, Eun Kyung; Kang, Nam Kyu; Jeon, Seungjib; Kwon, Sohee; Shin, Won-Sub; Lee, Bongsoo; Hwangbo, Kwon; Kim, Jungeun; Ye, Sung Hyeok; Yun, Jae-Young; Seo, Hogyun; Oh, Hee-Mock; Kim, Kyung-Jin; Kim, Jin-Soo; Jeong, Won-Joong; Chang, Yong Keun; Jeong, Byeong-Ryool

    2016-06-13

    Genome editing is crucial for genetic engineering of organisms for improved traits, particularly in microalgae due to the urgent necessity for the next generation biofuel production. The most advanced CRISPR/Cas9 system is simple, efficient and accurate in some organisms; however, it has proven extremely difficult in microalgae including the model alga Chlamydomonas. We solved this problem by delivering Cas9 ribonucleoproteins (RNPs) comprising the Cas9 protein and sgRNAs to avoid cytotoxicity and off-targeting associated with vector-driven expression of Cas9. We obtained CRISPR/Cas9-induced mutations at three loci including MAA7, CpSRP43 and ChlM, and targeted mutagenic efficiency was improved up to 100 fold compared to the first report of transgenic Cas9-induced mutagenesis. Interestingly, we found that unrelated vectors used for the selection purpose were predominantly integrated at the Cas9 cut site, indicative of NHEJ-mediated knock-in events. As expected with Cas9 RNPs, no off-targeting was found in one of the mutagenic screens. In conclusion, we improved the knockout efficiency by using Cas9 RNPs, which opens great opportunities not only for biological research but also industrial applications in Chlamydomonas and other microalgae. Findings of the NHEJ-mediated knock-in events will allow applications of the CRISPR/Cas9 system in microalgae, including "safe harboring" techniques shown in other organisms.

  2. CRISPR/Cas9-induced knockout and knock-in mutations in Chlamydomonas reinhardtii

    PubMed Central

    Shin, Sung-Eun; Lim, Jong-Min; Koh, Hyun Gi; Kim, Eun Kyung; Kang, Nam Kyu; Jeon, Seungjib; Kwon, Sohee; Shin, Won-Sub; Lee, Bongsoo; Hwangbo, Kwon; Kim, Jungeun; Ye, Sung Hyeok; Yun, Jae-Young; Seo, Hogyun; Oh, Hee-Mock; Kim, Kyung-Jin; Kim, Jin-Soo; Jeong, Won-Joong; Chang, Yong Keun; Jeong, Byeong-ryool

    2016-01-01

    Genome editing is crucial for genetic engineering of organisms for improved traits, particularly in microalgae due to the urgent necessity for the next generation biofuel production. The most advanced CRISPR/Cas9 system is simple, efficient and accurate in some organisms; however, it has proven extremely difficult in microalgae including the model alga Chlamydomonas. We solved this problem by delivering Cas9 ribonucleoproteins (RNPs) comprising the Cas9 protein and sgRNAs to avoid cytotoxicity and off-targeting associated with vector-driven expression of Cas9. We obtained CRISPR/Cas9-induced mutations at three loci including MAA7, CpSRP43 and ChlM, and targeted mutagenic efficiency was improved up to 100 fold compared to the first report of transgenic Cas9-induced mutagenesis. Interestingly, we found that unrelated vectors used for the selection purpose were predominantly integrated at the Cas9 cut site, indicative of NHEJ-mediated knock-in events. As expected with Cas9 RNPs, no off-targeting was found in one of the mutagenic screens. In conclusion, we improved the knockout efficiency by using Cas9 RNPs, which opens great opportunities not only for biological research but also industrial applications in Chlamydomonas and other microalgae. Findings of the NHEJ-mediated knock-in events will allow applications of the CRISPR/Cas9 system in microalgae, including “safe harboring” techniques shown in other organisms. PMID:27291619

  3. Spontaneous Dominant Mutations in Chlamydomonas Highlight Ongoing Evolution by Gene Diversification

    PubMed Central

    Boulouis, Alix; Drapier, Dominique; Razafimanantsoa, Hélène; Wostrikoff, Katia; Tourasse, Nicolas J.; Pascal, Kevin; Girard-Bascou, Jacqueline; Vallon, Olivier; Wollman, Francis-André; Choquet, Yves

    2015-01-01

    We characterized two spontaneous and dominant nuclear mutations in the unicellular alga Chlamydomonas reinhardtii, ncc1 and ncc2 (for nuclear control of chloroplast gene expression), which affect two octotricopeptide repeat (OPR) proteins encoded in a cluster of paralogous genes on chromosome 15. Both mutations cause a single amino acid substitution in one OPR repeat. As a result, the mutated NCC1 and NCC2 proteins now recognize new targets that we identified in the coding sequences of the chloroplast atpA and petA genes, respectively. Interaction of the mutated proteins with these targets leads to transcript degradation; however, in contrast to the ncc1 mutation, the ncc2 mutation requires on-going translation to promote the decay of the petA mRNA. Thus, these mutants reveal a mechanism by which nuclear factors act on chloroplast mRNAs in Chlamydomonas. They illustrate how diversifying selection can allow cells to adapt the nuclear control of organelle gene expression to environmental changes. We discuss these data in the wider context of the evolution of regulation by helical repeat proteins. PMID:25804537

  4. Atomic resolution modeling of the ferredoxin:[FeFe] hydrogenase complex from Chlamydomonas reinhardtii.

    PubMed

    Chang, Christopher H; King, Paul W; Ghirardi, Maria L; Kim, Kwiseon

    2007-11-01

    The [FeFe] hydrogenases HydA1 and HydA2 in the green alga Chlamydomonas reinhardtii catalyze the final reaction in a remarkable metabolic pathway allowing this photosynthetic organism to produce H(2) from water in the chloroplast. A [2Fe-2S] ferredoxin is a critical branch point in electron flow from Photosystem I toward a variety of metabolic fates, including proton reduction by hydrogenases. To better understand the binding determinants involved in ferredoxin:hydrogenase interactions, we have modeled Chlamydomonas PetF1 and HydA2 based on amino-acid sequence homology, and produced two promising electron-transfer model complexes by computational docking. To characterize these models, quantitative free energy calculations at atomic resolution were carried out, and detailed analysis of the interprotein interactions undertaken. The protein complex model we propose for ferredoxin:HydA2 interaction is energetically favored over the alternative candidate by 20 kcal/mol. This proposed model of the electron-transfer complex between PetF1 and HydA2 permits a more detailed view of the molecular events leading up to H(2) evolution, and suggests potential mutagenic strategies to modulate electron flow to HydA2.

  5. High-Throughput Robotically Assisted Isolation of Temperature-sensitive Lethal Mutants in Chlamydomonas reinhardtii

    PubMed Central

    Breker, Michal; Lieberman, Kristi; Tulin, Frej; Cross, Frederick R.

    2016-01-01

    Systematic identification and characterization of genetic perturbations have proven useful to decipher gene function and cellular pathways. However, the conventional approaches of permanent gene deletion cannot be applied to essential genes. We have pioneered a unique collection of ~70 temperature-sensitive (ts) lethal mutants for studying cell cycle regulation in the unicellular green algae Chlamydomonas reinhardtii1. These mutations identify essential genes, and the ts alleles can be conditionally inactivated by temperature shift, providing valuable tools to identify and analyze essential functions. Mutant collections are much more valuable if they are close to comprehensive, since scattershot collections can miss important components. However, this requires the efficient collection of a large number of mutants, especially in a wide-target screen. Here, we describe a robotics-based pipeline for generating ts lethal mutants and analyzing their phenotype in Chlamydomonas. This technique can be applied to any microorganism that grows on agar. We have collected over 3000 ts mutants, probably including mutations in most or all cell-essential pathways, including about 200 new candidate cell cycle mutations. Subsequent molecular and cellular characterization of these mutants should provide new insights in plant cell biology; a comprehensive mutant collection is an essential prerequisite to ensure coverage of a broad range of biological pathways. These methods are integrated with downstream genetics and bioinformatics procedures for efficient mapping and identification of the causative mutations that are beyond the scope of this manuscript. PMID:28060315

  6. Membrane-associated polypeptides induced in Chlamydomonas by limiting CO sub 2 concentrations

    SciTech Connect

    Spalding, M.H.; Jeffrey, M. )

    1989-01-01

    Chlamydomonas reinhardtii and other unicellular green algae have a high apparent affinity for CO{sub 2}, little O{sub 2} inhibition of photosynthesis, and reduced photorespiration. These characteristics result from operation of a CO{sub 2}-concentrating system. The CO{sub 2}-concentrating system involves active inorganic carbon transport and is under environmental control. Cells grown at limiting CO{sub 2} concentrations have inorganic carbon transport activity, but cells grown at 5% CO{sub 2} do not. Four membrane-associated polypeptides (M{sub r}, 19, 21, 35, and 36 kilodaltons) have been identified which either appear or increase in abundance during adaptation to limiting CO{sub 2} concentrations. The appearance of two of the polypeptides occurs over roughly the same time course as the appearance of the CO{sub 2}-concentrating system activity in response to CO{sub 2} limitation.

  7. Ethanol stimulates phospholipid turnover and inositol 1,4,5-trisphosphate production in Chlamydomonas eugametos gametes.

    PubMed

    Musgrave, A; Kuin, H; Jongen, M; de Wildt, P; Schuring, F; Klerk, H; van den Ende, H

    1992-02-01

    Alcohols induce mating-structure activation in Chlamydomonas eugametos gametes. From the effect of ethanol on the (32)P-labelling of polyphosphoinositides, we conclude that the synthesis of these lipids is stimulated. Biologically inactive concentrations of ethanol (<6%) had no effect on synthesis, but 6-8% ethanol stimulated synthesis for upto 60 min. The (32)P incorporated into polyphosphoinositides and phosphatidic acid during ethanol treatment was readily chased out when 1 mM unlabelled Na3PO4 was added. Using a binding assay for inositol 1,4,5-trisphosphate, we show that the production of this phospholipid constituent is dramatically increased after ethanol treatment. This effect, coupled to a rise in intracellular calcium concentration, could explain gamete activation. The significance of these results in explaining other ethanol-induced phenomena in algae is discussed.

  8. [LIGHT-DEPENDENT SYNTHESIS OF CELL MEMBRANES IN THE Brc-1 MUTANT OF CHLAMYDOMONAS REINHARDTII].

    PubMed

    Semenova, G A; Chekunova, E M; Ladygin, V G

    2015-01-01

    The structural organization of cells of the Brc-1 mutant of the unicellular green algae Chlamydomonas reinhardtii grown in the light and in the dark has been studied. The Brc-1 mutant contains the brc-1 mutation in the nucleus gene LTS3. In the light, all membrane structures in mutant cells form normally and are well developed. In the dark under heterotrophic conditions, the mutant cells grew and divided well, however, all its cell membranes: plasmalemma, tonoplast, mitochondrial membranes, membranes of the nucleus shell and chloroplast, thylakoids, and the membranes of dictiosomes of the Golgi apparatus were not detected. In the dark under heterotrophic conditions, mutant cells well grow and divide. It were shown that a short-term (1-10 min) exposure of Brc-1 mutant cells to light leads to the restoration of all above-mentioned membrane structures. Possible reasons for the alterations of membrane structures are discussed.

  9. Epigenetic silencing of a foreign gene in nuclear transformants of Chlamydomonas.

    PubMed Central

    Cerutti, H; Johnson, A M; Gillham, N W; Boynton, J E

    1997-01-01

    The unstable expression of introduced genes poses a serious problem for the application of transgenic technology in plants. In transformants of the unicellular green alga Chlamydomonas reinhardtii, expression of a eubacterial aadA gene, conferring spectinomycin resistance, is transcriptionally suppressed by a reversible epigenetic mechanism(s). Variations in the size and frequency of colonies surviving on different concentrations of spectinomycin as well as the levels of transcriptional activity of the introduced transgene(s) suggest the existence of intermediate expression states in genetically identical cells. Gene silencing does not correlate with methylation of the integrated DNA and does not involve large alterations in its chromatin structure, as revealed by digestion with restriction endonucleases and DNase I. Transgene repression is enhanced by lower temperatures, similar to position effect variegation in Drosophila. By analogy to epigenetic phenomena in several eukaryotes, our results suggest a possible role for (hetero)chromatic chromosomal domains in transcriptional inactivation. PMID:9212467

  10. Molecular characterization of a zygote wall protein: an extensin-like molecule in Chlamydomonas reinhardtii.

    PubMed Central

    Woessner, J P; Goodenough, U W

    1989-01-01

    The green alga Chlamydomonas reinhardtii elaborates two biochemically and morphologically distinct cell walls during its life cycle: one surrounds the vegetative and gametic cell and the other encompasses the zygote. Hydroxyproline-rich glycoproteins (HRGPs) constitute a major component of both walls. We describe the isolation and characterization of a zygote-specific gene encoding a wall HRGP. The derived amino acid sequence of this algal HRGP is similar to those of higher plant extensins, rich in proline and serine residues and possessing repeating amino acid motifs, notably X(Pro)3 and (Ser-Pro)n. Antiserum against this zygote wall protein detected common epitopes in several other zygote polypeptides, at least one of which is also encoded by a zygote-specific gene. We conclude that there is one set of HRGP wall genes expressed only in zygotes and another set that is specific to vegetative and gametic cells. PMID:2535530

  11. Chlamydomonas swims with two "gears" in a eukaryotic version of run-and-tumble locomotion.

    PubMed

    Polin, Marco; Tuval, Idan; Drescher, Knut; Gollub, J P; Goldstein, Raymond E

    2009-07-24

    The coordination of eukaryotic flagella is essential for many of the most basic processes of life (motility, sensing, and development), yet its emergence and regulation and its connection to locomotion are poorly understood. Previous studies show that the unicellular alga Chlamydomonas, widely regarded as an ideal system in which to study flagellar biology, swims forward by the synchronous action of its two flagella. Using high-speed imaging over long intervals, we found a richer behavior: A cell swimming in the dark stochastically switches between synchronous and asynchronous flagellar beating. Three-dimensional tracking shows that these regimes lead, respectively, to nearly straight swimming and to abrupt large reorientations, which yield a eukaryotic version of the "run-and-tumble" motion of peritrichously flagellated bacteria.

  12. Functional and Spectroscopic Characterization of Chlamydomonas reinhardtii Truncated Hemoglobins

    PubMed Central

    Droghetti, Enrica; Tundo, Grazia R.; Sanz-Luque, Emanuel; Polticelli, Fabio; Visca, Paolo; Smulevich, Giulietta; Ascenzi, Paolo; Coletta, Massimo

    2015-01-01

    The single-cell green alga Chlamydomonas reinhardtii harbors twelve truncated hemoglobins (Cr-TrHbs). Cr-TrHb1-1 and Cr-TrHb1-8 have been postulated to be parts of the nitrogen assimilation pathway, and of a NO-dependent signaling pathway, respectively. Here, spectroscopic and reactivity properties of Cr-TrHb1-1, Cr-TrHb1-2, and Cr-TrHb1-4, all belonging to clsss 1 (previously known as group N or group I), are reported. The ferric form of Cr-TrHb1-1, Cr-TrHb1-2, and Cr-TrHb1-4 displays a stable 6cLS heme-Fe atom, whereas the hexa-coordination of the ferrous derivative appears less strongly stabilized. Accordingly, kinetics of azide binding to ferric Cr-TrHb1-1, Cr-TrHb1-2, and Cr-TrHb1-4 are independent of the ligand concentration. Conversely, kinetics of CO or NO2− binding to ferrous Cr-TrHb1-1, Cr-TrHb1-2, and Cr-TrHb1-4 are ligand-dependent at low CO or NO2− concentrations, tending to level off at high ligand concentrations, suggesting the presence of a rate-limiting step. In agreement with the different heme-Fe environments, the pH-dependent kinetics for CO and NO2−binding to ferrous Cr-TrHb1-1, Cr-TrHb1-2, and Cr-TrHb1-4 are characterized by different ligand-linked protonation events. This raises the question of whether the simultaneous presence in C. reinhardtii of multiple TrHb1s may be related to different regulatory roles. PMID:25993270

  13. Nuclear transformation of Chlamydomonas reinhardtii with silicon carbide fibers

    SciTech Connect

    Dunahay, T.G. )

    1992-01-01

    Efficient nuclear transformation of cell wall-deficient strains of the green alga Chlamydomonas reinhardtii can be accomplished by vortexing the cells in the presence of glass beads and polyethylene glycol (Kindle 1990 PNAS 87:1228). Intact (walled) cells can also be transformed using this protocol, but at very low efficiencies. Two recent reports have described the use of silicon carbide fibers to mediate DNA entry into plant suspension cells (Kaeppler et al. 1990 Plant Cell Rep. 9:414; Asano et al. 1991 Plant Sci. 79:247). The author has found that nuclear transformation efficiencies of walled cells of C. reinhardtii can be increased 3 to 10 fold by vortexing the cells in the presence of silicon carbide fibers and PEG. Using a modification of the glass bead transformation procedure, the wild-type nitrate reductase structural gene was used to complement a NR-deficient mutant of C. reinhardtii, nit-1-305. The transformation efficiency increased with longer vortexing times, although the absolute number of transformants varied between experiments, ranging from 10 to 40 transformants per 10[sup 7] cells. In contrast to vortexing with glass beads, cell viability was very high, with greater than 80% cell survival even after vortexing for 10 minutes. Neither cell death nor transformation efficiency increased when cell wall-deficient mutants (cw15 nit-1-305) were used as compared to intact cells. Experiments are in progress to test the applicability of silicon carbide-mediated transformation to other algal strains for which cell wall mutants or protoplasting procedures are unavailabile.

  14. Alkaloids in Marine Algae

    PubMed Central

    Güven, Kasım Cemal; Percot, Aline; Sezik, Ekrem

    2010-01-01

    This paper presents the alkaloids found in green, brown and red marine algae. Algal chemistry has interested many researchers in order to develop new drugs, as algae include compounds with functional groups which are characteristic from this particular source. Among these compounds, alkaloids present special interest because of their pharmacological activities. Alkaloid chemistry has been widely studied in terrestrial plants, but the number of studies in algae is insignificant. In this review, a detailed account of macro algae alkaloids with their structure and pharmacological activities is presented. The alkaloids found in marine algae may be divided into three groups: 1. Phenylethylamine alkaloids, 2. Indole and halogenated indole alkaloids, 3. Other alkaloids. PMID:20390105

  15. Synergism between Inositol Polyphosphates and TOR Kinase Signaling in Nutrient Sensing, Growth Control, and Lipid Metabolism in Chlamydomonas[OPEN

    PubMed Central

    Evans, Bradley S.; Li, Jia; Liu, Yu; Diamond, Spencer

    2016-01-01

    The networks that govern carbon metabolism and control intracellular carbon partitioning in photosynthetic cells are poorly understood. Target of Rapamycin (TOR) kinase is a conserved growth regulator that integrates nutrient signals and modulates cell growth in eukaryotes, though the TOR signaling pathway in plants and algae has yet to be completely elucidated. We screened the unicellular green alga Chlamydomonas reinhardtii using insertional mutagenesis to find mutants that conferred hypersensitivity to the TOR inhibitor rapamycin. We characterized one mutant, vip1-1, that is predicted to encode a conserved inositol hexakisphosphate kinase from the VIP family that pyrophosphorylates phytic acid (InsP6) to produce the low abundance signaling molecules InsP7 and InsP8. Unexpectedly, the rapamycin hypersensitive growth arrest of vip1-1 cells was dependent on the presence of external acetate, which normally has a growth-stimulatory effect on Chlamydomonas. vip1-1 mutants also constitutively overaccumulated triacylglycerols (TAGs) in a manner that was synergistic with other TAG inducing stimuli such as starvation. vip1-1 cells had reduced InsP7 and InsP8, both of which are dynamically modulated in wild-type cells by TOR kinase activity and the presence of acetate. Our data uncover an interaction between the TOR kinase and inositol polyphosphate signaling systems that we propose governs carbon metabolism and intracellular pathways that lead to storage lipid accumulation. PMID:27600537

  16. Synergism between inositol polyphosphates and TOR kinase signaling in nutrient sensing, growth control and lipid metabolism in Chlamydomonas.

    PubMed

    Couso, Inmaculada; Evans, Bradley; Li, Jia; Liu, Yu; Ma, Fangfang; Diamond, Spencer; Allen, Doug K; Umen, James G

    2016-09-06

    The networks that govern carbon metabolism and control intracellular carbon partitioning in photosynthetic cells are poorly understood. Target of rapamycin (TOR) kinase is a conserved growth regulator that integrates nutrient signals and modulates cell growth in eukaryotes, though the TOR signaling pathway in plants and algae has yet to be completely elucidated. We screened the unicellular green alga Chlamydomonas using insertional mutagenesis to find mutants that conferred hypersensitivity to the TOR inhibitor rapamycin. We characterized one mutant, vip1-1, that is predicted to encode a conserved inositol hexakisphosphate kinase from the VIP family that pyrophosphorylates phytic acid (InsP6) to produce the low abundance signaling molecules InsP7 and InsP8. Unexpectedly, the rapamycin hypersensitive growth arrest of vip1-1 cells was dependent on the presence of external acetate, which normally has a growth-stimulatory effect on Chlamydomonas. vip1-1 mutants also constitutively over-accumulated triacylglycerols (TAGs) in a manner that was synergistic with other TAG inducing stimuli such as starvation. vip1-1 cells had reduced InsP7 and InsP8, both of which are dynamically modulated in wild-type cells by TOR kinase activity and the presence of acetate. Our data uncover an interaction between the TOR kinase and inositol polyphosphate signaling systems that we propose governs carbon metabolism and intracellular pathways that lead to storage lipid accumulation.

  17. High iron content and bioavailability in humans from four species of marine algae.

    PubMed

    García-Casal, Maria N; Pereira, Ana C; Leets, Irene; Ramírez, José; Quiroga, Maria F

    2007-12-01

    Searching for economical, nonconventional sources of iron is important in underdeveloped countries to combat iron deficiency and anemia. Our objective was to study iron, vitamin C, and phytic acid composition and also iron bioavailability from 4 species of marine algae included in a rice-based meal. Marine algae (Ulva sp, Sargassum sp, Porphyra sp, and Gracilariopsis sp) were analyzed for monthly variations in iron and for ascorbic acid and phytic acid concentrations. A total of 96 subjects received rice-based meals containing the 4 species of marine algae in different proportions, raw or cooked. All meals contained radioactive iron. Absorption was evaluated by calculating the radioactive iron incorporation in subjects' blood. Iron concentrations in algae were high and varied widely, depending on the species and time of year. The highest iron concentrations were found in Sargassum (157 mg/100 g) and Gracilariopsis (196 mg/100 g). Phytates were not detected in the algae and ascorbic acid concentration fluctuated between 38 microg/g dry weight (Ulva) and 362 microg/g dry weight (Sargassum). Algae significantly increased iron absorption in rice-based meals. Cooking did not affect iron absorption compared with raw algae. Results indicate that Ulva sp, Sargassum sp, Porphyra sp, and Gracilariopsis sp are good sources of ascorbic acid and bioavailable iron. The percentage of iron absorption was similar among all algae tested, although Sargassum sp resulted in the highest iron intake. Based on these results, and on the high reproduction rates of algae during certain seasons, promoting algae consumption in some countries could help to improve iron nutrition.

  18. Efficient H2 production via Chlamydomonas reinhardtii.

    PubMed

    Esquível, Maria G; Amaro, Helena M; Pinto, Teresa S; Fevereiro, Pedro S; Malcata, F Xavier

    2011-12-01

    Molecular hydrogen (H(2)) obtained from biological sources provides an alternative to bulk chemical processes that is moving towards large-scale, economical generation of clean fuel for automotive engines. This opinion article examines recent improvements in H(2) production by wild and mutant strains of Chlamydomonas reinhardtii - the green microalga currently considered the best eukaryotic H(2) producer. Here, we review various aspects of genetic and metabolic engineering of C. reinhardtii, as well as of process engineering. Additionally, we lay out possible scenarios that would lead to more efficient research approaches in the near future, as part of a consistent strategy for sustainable biohydrogen supply.

  19. Functional determinants in transit sequences: import and partial maturation by vascular plant chloroplasts of the ribulose-1,5- bisphosphate carboxylase small subunit of Chlamydomonas

    PubMed Central

    1985-01-01

    The precursor of the ribulose-1,5-bisphosphate carboxylase small subunit and other proteins from Chlamydomonas reinhardtii are efficiently transported into chloroplasts isolated from spinach and pea. Thus, similar determinants specify precursor-chloroplast interactions in the alga and vascular plants. Removal of all or part of its transit sequence was found to block import of the algal small subunit into isolated chloroplasts. Comparison of available sequences revealed a nine amino acid segment conserved in the transit sequences of all small subunit precursors. A protease in the vascular plant chloroplasts recognized this region in the Chlamydomonas precursor and produced an intermediate form of the small subunit. We propose that processing of the small subunit precursor involves at least two proteolytic events; only one of these has been evolutionarily conserved. PMID:3965471

  20. Taxonomic revision of oil-producing green algae, Chlorococcum oleofaciens (Volvocales, Chlorophyceae), and its relatives.

    PubMed

    Kawasaki, Yuriko; Nakada, Takashi; Tomita, Masaru

    2015-10-01

    Historically, species in Volvocales were classified based primarily on morphology. Although the taxonomy of Chlamydomonas has been re-examined using a polyphasic approach including molecular phylogeny, that of Chlorococcum (Cc.), the largest coccoid genus in Volvocales, has yet to be reexamined. Six species thought to be synonymous with the oil-producing alga Cc. oleofaciens were previously not confirmed by molecular phylogeny. In this study, seven authentic strains of Cc. oleofaciens and its putative synonyms, along with 11 relatives, were examined based on the phylogeny of the 18S ribosomal RNA (rRNA) gene, comparisons of secondary structures of internal transcribed spacer 1 (ITS1) and ITS2 rDNA, and morphological observations by light microscopy. Seven 18S rRNA types were recognized among these strains and three were distantly related to Cc. oleofaciens. Comparisons of ITS rDNA structures suggested possible separation of the remaining four types into different species. Shapes of vegetative cells, thickness of the cell walls in old cultures, the size of cells in old cultures, and stigma morphology of zoospores also supported the 18S rRNA grouping. Based on these results, the 18 strains examined were reclassified into seven species. Among the putative synonyms, synonymy of Cc. oleofaciens, Cc. croceum, and Cc. granulosum was confirmed, and Cc. microstigmatum, Cc. rugosum, Cc. aquaticum, and Cc. nivale were distinguished from Cc. oleofaciens. Furthermore, another related strain is described as a new species, Macrochloris rubrioleum sp. nov.

  1. Phytotoxicity, bioaccumulation and degradation of isoproturon in green algae.

    PubMed

    Bi, Yan Fang; Miao, Shan Shan; Lu, Yi Chen; Qiu, Chong Bin; Zhou, You; Yang, Hong

    2012-12-01

    Isoproturon (IPU) is a pesticide used for protection of land crops from weed or pathogen attack. Recent survey shows that IPU has been detected as a contaminant in aquatic systems and may have negative impact on aquatic organisms. To understand the phytotoxicity and potential accumulation and degradation of IPU in algae, a comprehensive study was performed with the green alga Chlamydomonas reinhardtii. Algae exposed to 5-50 μg L(-1) IPU for 3d displayed progressive inhibition of cell growth and reduced chlorophyll fluorescence. Time-course experiments with 25 μg L(-1) IPU for 6d showed similar growth responses. The 72 h EC50 value for IPU was 43.25 μg L(-1), NOEC was 5 μg L(-1) and LOEC was 15 μg L(-1). Treatment with IPU induced oxidative stress. This was validated by a group of antioxidant enzymes, whose activities were promoted by IPU exposure. The up-regulation of several genes coding for the enzymes confirmed the observation. IPU was shown to be readily accumulated by C. reinhardtii. However, the alga showed a weak ability to degrade IPU accumulated in its cells, which was best presented at the lower concentration (5 μg L(-1)) of IPU in the medium. The imbalance of accumulation and degradation of IPU may be the cause that resulted in the detrimental growth and cellular damage.

  2. Expression and assembly of a fully active antibody in algae

    NASA Astrophysics Data System (ADS)

    Mayfield, Stephen P.; Franklin, Scott E.; Lerner, Richard A.

    2003-01-01

    Although combinatorial antibody libraries have solved the problem of access to large immunological repertoires, efficient production of these complex molecules remains a problem. Here we demonstrate the efficient expression of a unique large single-chain (lsc) antibody in the chloroplast of the unicellular, green alga, Chlamydomonas reinhardtii. We achieved high levels of protein accumulation by synthesizing the lsc gene in chloroplast codon bias and by driving expression of the chimeric gene using either of two C. reinhardtii chloroplast promoters and 5' and 3' RNA elements. This lsc antibody, directed against glycoprotein D of the herpes simplex virus, is produced in a soluble form by the alga and assembles into higher order complexes in vivo. Aside from dimerization by disulfide bond formation, the antibody undergoes no detectable posttranslational modification. We further demonstrate that accumulation of the antibody can be modulated by the specific growth regime used to culture the alga, and by the choice of 5' and 3' elements used to drive expression of the antibody gene. These results demonstrate the utility of alga as an expression platform for recombinant proteins, and describe a new type of single chain antibody containing the entire heavy chain protein, including the Fc domain.

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

    PubMed

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

    2015-10-01

    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 herbicides paraquat and diuron, suggesting that the fast physiological and biochemical defence mechanisms that conferred tolerance of algae towards higher UVR levels were related to singlet oxygen defence. The presented study suggests that knowledge of the molecular toxicity mechanisms of chemicals, rather than their general physiological target, is needed in order to predict co-tolerance between environmental and chemical stressors.

  4. [An experiment with Chlamydomonas reinhardtii on the Kosmos-2044 biosatellite].

    PubMed

    Gavrilova, O V; Gabova, A V; Goriainova, L N; Filatova, E V

    1992-01-01

    Space experiment with Chlamydomonas reinhardtii demonstrated that the microgravity effects were noted in Chlamydomonas at both cellular and population levels: in space the cell size is increased, stage of active growth of the culture is extended, it contains the juvenile vegetative motile cells in greater quantities. Ultrastructural analysis indicated that in microgravity the changes in shape, structure and distribution of intracellular organelles and in volume ratio of organelles and cytoplasma are absent. Chlamydomonas data are in line with the results of the Infusoria and Chlorella experiments.

  5. Paternal inheritance of mitochondria in Chlamydomonas.

    PubMed

    Nakamura, Soichi

    2010-03-01

    To analyze mitochondrial DNA (mtDNA)inheritance, differences in mtDNA between Chlamydomonas reinhardtii and Chlamydomonas smithii, respiration deficiency and antibiotic resistance were used to distinguish mtDNA origins. The analyses indicated paternal inheritance. However, these experiments raised questions regarding whether paternal inheritance occurred normally.Mitochondrial nucleoids were observed in living zygotes from mating until 3 days after mating and then until progeny formation. However, selective disappearance of nucleoids was not observed. Subsequently, experimental serial backcrosses between the two strains demonstrated strict paternal inheritance. The fate of mt+ and mt- mtDNA was followed using the differences in mtDNA between the two strains. The slow elimination of mt+ mtDNA through zygote maturation in darkness was observed, and later the disappearance of mt+ mtDNA was observed at the beginning of meiosis. To explain the different fates of mtDNA, methylation status was investigated; however, no methylation was detected. Variously constructed diploid cells showed biparental inheritance. Thus, when the mating process occurs normally, paternal inheritance occurs. Mutations disrupting mtDNA inheritance have not yet been isolated. Mutations that disrupt maternal inheritance of chloroplast DNA (cpDNA) do not disrupt inheritance of mtDNA. The genes responsible for mtDNA inheritance are different from those of chloroplasts.

  6. The Role of Glycerol and Inorganic Ions in Osmoregulatory Responses of the Euryhaline Flagellate Chlamydomonas pulsatilla Wollenweber 1

    PubMed Central

    Ahmad, Iftikhar; Hellebust, Johan A.

    1986-01-01

    The green euryhaline flagellate Chlamydomonas pulsatilla Wollenweber, isolated from a coastal marine environment, was grown exponentially over the salinity range of 10 to 200% artificial seawater (ASW). The cellular volume and aqueous space of the alga, measured by [14C] mannitol and 3H2O tracer analyses of centrifuged cell pellets, ranged between 2.3 and 3.1 picoliters and between 1.5 and 2.1 picoliters, respectively. The nonaqueous space determined in those analyses (28-35%) was consistent with the cell composition of the alga. The glycerol content of the alga increased almost linearly with increasing salinity; its contribution to intracellular osmolality at 200% ASW was about 57%. The contribution of amino acids and soluble carbohydrates to the cell osmotic balance was small. Intracellular ion concentrations determined by analyzing centrifuged cell pellets of known [14C]mannitol space by atomic absorption spectrophotometry, and by neutron activation analyses of washed cells were similar. At 10% ASW, potassium and magnesium were the major cations, and chloride and phosphate were the major anions. The sodium and chloride content of the alga increased with increasing salinity; at 200% ASW the intracellular concentration of both sodium and chloride was about 400 millimolar. The intracellular osmolality (πint) matched closely the external osmolality (πext) over the entire salinity range except at 10% ASW where πint exceeded πext by 120 to 270 milliosmoles per kilogram H2O. PMID:16665042

  7. Oleosin of subcellular lipid droplets evolved in green algae.

    PubMed

    Huang, Nan-Lan; Huang, Ming-Der; Chen, Tung-Ling L; Huang, Anthony H C

    2013-04-01

    In primitive and higher plants, intracellular storage lipid droplets (LDs) of triacylglycerols are stabilized with a surface layer of phospholipids and oleosin. In chlorophytes (green algae), a protein termed major lipid-droplet protein (MLDP) rather than oleosin on LDs was recently reported. We explored whether MLDP was present directly on algal LDs and whether algae had oleosin genes and oleosins. Immunofluorescence microscopy revealed that MLDP in the chlorophyte Chlamydomonas reinhardtii was associated with endoplasmic reticulum subdomains adjacent to but not directly on LDs. In C. reinhardtii, low levels of a transcript encoding an oleosin-like protein (oleolike) in zygotes-tetrads and a transcript encoding oleosin in vegetative cells transferred to an acetate-enriched medium were found in transcriptomes and by reverse transcription-polymerase chain reaction. The C. reinhardtii LD fraction contained minimal proteins with no detectable oleolike or oleosin. Several charophytes (advanced green algae) possessed low levels of transcripts encoding oleosin but not oleolike. In the charophyte Spirogyra grevilleana, levels of oleosin transcripts increased greatly in cells undergoing conjugation for zygote formation, and the LD fraction from these cells contained minimal proteins, two of which were oleosins identified via proteomics. Because the minimal oleolike and oleosins in algae were difficult to detect, we tested their subcellular locations in Physcomitrella patens transformed with the respective algal genes tagged with a Green Fluorescent Protein gene and localized the algal proteins on P. patens LDs. Overall, oleosin genes having weak and cell/development-specific expression were present in green algae. We present a hypothesis for the evolution of oleosins from algae to plants.

  8. Basis for the Resistance of Several Algae to Microbial Decomposition

    PubMed Central

    Gunnison, Douglas; Alexander, Martin

    1975-01-01

    The basis for the resistance of certain algae to microbial decomposition in natural waters was investigated using Pediastrum duplex, Staurastrum sp., and Fischerella muscicola as test organisms. Enzyme preparations previously found to convert susceptible algae into spheroplasts had no such effect on the resistant species, although glucose and galacturonic acid were released from P. duplex walls. Little protein or lipid but considerable carbohydrate was found in the walls of the refractory organisms, but resistance was not correlated with the presence of a unique sugar monomer. A substance present in Staurastrum sp. walls was characterized as lignin or lignin-like on the basis of its extraction characteristics, infrared spectrum, pyrolysis pattern, and content of an aromatic building block. Sporopollenin was found in P. duplex, and cellulose in Staurastrum sp. Cell walls of the algae were fractionated, and the fractions least susceptible to microbial degradation were the sporopollenin of P. duplex, the polyaromatic component of Staurastrum sp., and two F. muscicola fractions containing several sugar monomers. The sporopollenin content of P. duplex, the content of lignin or a related constituent of Staurastrum sp., and the resistance of the algae to microbial attack increased with age. It is suggested that resistance results from the presence of sporopollenin in P. duplex, a lignin-like material in Staurastrum sp., and possibly heteropolysaccharides in F. muscicola. PMID:808166

  9. Analysis of an essential carotenogenic enzyme: ζ-carotene desaturase from unicellular Alga Dunaliella salina.

    PubMed

    Ye, Zhi-Wei; Jiang, Jian-Guo

    2010-11-10

    The green alga Dunaliella has become a valuable model organism for understanding the interesting mechanism of massive carotenoid accumulation. Previously, DNA sequences of several carotenogenic enzymes were obtained from Dunaliella. In this study, the cDNA of zds was isolated from Dunaliella salina using a polymerase chain reaction approach. The full-length cDNA sequence was 2178 base pairs (bp) containing a 1731 bp putative open reading frame which coded a 576 amino acid deduced polypeptide whose molecular weight was 63.9 kDa computationally. A complete homologous search displayed that the nucleotide and putative protein sequence have sequence identities of 69% and 66% with those of green alga Chlamydomonas reinhardtii, respectively. It was predicted that this ζ-carotene desaturase (Zds) may be located in the chloroplast of D. salina. Phylogenetic analysis demonstrated that the D. salina Zds had a closer relationship with the Zds of algae and higher plants than with those of other species.

  10. Algae after dark: mechanisms to cope with anoxic/hypoxic conditions.

    PubMed

    Yang, Wenqiang; Catalanotti, Claudia; Wittkopp, Tyler M; Posewitz, Matthew C; Grossman, Arthur R

    2015-05-01

    Chlamydomonas reinhardtii is a unicellular, soil-dwelling (and aquatic) green alga that has significant metabolic flexibility for balancing redox equivalents and generating ATP when it experiences hypoxic/anoxic conditions. The diversity of pathways available to ferment sugars is often revealed in mutants in which the activities of specific branches of fermentative metabolism have been eliminated; compensatory pathways that have little activity in parental strains under standard laboratory fermentative conditions are often activated. The ways in which these pathways are regulated and integrated have not been extensively explored. In this review, we primarily discuss the intricacies of dark anoxic metabolism in Chlamydomonas, but also discuss aspects of dark oxic metabolism, the utilization of acetate, and the relatively uncharacterized but critical interactions that link chloroplastic and mitochondrial metabolic networks.

  11. International Conference on the Cell and Molecular Biology of Chlamydomonas

    SciTech Connect

    Dr. Stephen Miller

    2010-06-10

    The 2010 Conference on the Cell and Molecular Biology of Chlamydomonas was held June 6-10 near Boston, MA, and attracted a record 273 participants, 146 from US labs, 10 from Canada, and the remainder from 18 other countries. The single-celled algal protist Chlamydomonas is a key research organism for many investigators, including those who study photosynthesis, cell motility, adaptation to environmental stresses, the evolution of multicellularity, and the production of biofuels. Chlamydomonas researchers gather every two years at a research conference to exchange methods, develop collaborative efforts, disseminate recent findings, and plan large-scale studies to improve the usefulness of this unique research organism. This conference provides the only opportunity for Chlamydomonas scientists who work on different research problems to meet face to face, and greatly speeds progress in their respective fields. An important function of these Chlamydomonas conferences is to promote and showcase the work of younger scientists, and to attract new investigators into the Chlamydomonas community. DOE award SC0004085 was used to offset the travel and registration costs for 18 young investigators, 9 of whom were women, including one African American. Most of these scientists would not have been able to attend the conference without DOE support. A total of 208 research presentations were made at the meeting, 80 talks (63 presented by students, postdocs, and pre-tenured faculty) and 128 posters. Cell motility and biofuels/metabolism were the best-represented research areas, with a total of 77 presentations. This fact underscores the growing importance of Chlamydomonas as a research and production tool in the rapidly expanding world of biofuels research. A total of 28 talks and posters were presented on the topics of photosynthesis and stress responses, which were among the next best-represented research areas. As at several recent Chlamydomonas meetings, important advances were

  12. Dynamic Changes in the Transcriptome and Methylome of Chlamydomonas reinhardtii throughout Its Life Cycle.

    PubMed

    Lopez, David; Hamaji, Takashi; Kropat, Janette; De Hoff, Peter; Morselli, Marco; Rubbi, Liudmilla; Fitz-Gibbon, Sorel; Gallaher, Sean D; Merchant, Sabeeha S; Umen, James; Pellegrini, Matteo

    2015-12-01

    The green alga Chlamydomonas reinhardtii undergoes gametogenesis and mating upon nitrogen starvation. While the steps involved in its sexual reproductive cycle have been extensively characterized, the genome-wide transcriptional and epigenetic changes underlying different life cycle stages have yet to be fully described. Here, we performed transcriptome and methylome sequencing to quantify expression and DNA methylation from vegetative and gametic cells of each mating type and from zygotes. We identified 361 gametic genes with mating type-specific expression patterns and 627 genes that are specifically induced in zygotes; furthermore, these sex-related gene sets were enriched for secretory pathway and alga-specific genes. We also examined the C. reinhardtii nuclear methylation map with base-level resolution at different life cycle stages. Despite having low global levels of nuclear methylation, we detected 23 hypermethylated loci in gene-poor, repeat-rich regions. We observed mating type-specific differences in chloroplast DNA methylation levels in plus versus minus mating type gametes followed by chloroplast DNA hypermethylation in zygotes. Lastly, we examined the expression of candidate DNA methyltransferases and found three, DMT1a, DMT1b, and DMT4, that are differentially expressed during the life cycle and are candidate DNA methylases. The expression and methylation data we present provide insight into cell type-specific transcriptional and epigenetic programs during key stages of the C. reinhardtii life cycle.

  13. Methanol-Promoted Lipid Remodelling during Cooling Sustains Cryopreservation Survival of Chlamydomonas reinhardtii

    PubMed Central

    Yang, Duanpeng; Li, Weiqi

    2016-01-01

    Cryogenic treatments and cryoprotective agents (CPAs) determine the survival rate of organisms that undergo cryopreservation, but their mechanisms of operation have not yet been characterised adequately. In particular, the way in which membrane lipids respond to cryogenic treatments and CPAs is unknown. We developed comparative profiles of the changes in membrane lipids among cryogenic treatments and between the CPAs dimethyl sulfoxide (DMSO) and methanol (MeOH) for the green alga Chlamydomonas reinhardtii. We found that freezing in liquid nitrogen led to a dramatic degradation of lipids, and that thawing at warm temperature (35°C) induced lipid remodelling. DMSO did not protect membranes, but MeOH significantly attenuated lipid degradation. The presence of MeOH during cooling (from 25°C to −55°C at a rate of 1°C/min) sustained the lipid composition to the extent that membrane integrity was maintained; this phenomenon accounts for successful cryopreservation. An increase in monogalactosyldiacylglycerol and a decrease in diacylglycerol were the major changes in lipid composition associated with survival rate, but there was no transformation between these lipid classes. Phospholipase D-mediated phosphatidic acid was not involved in freezing-induced lipid metabolism in C. reinhardtii. Lipid unsaturation changed, and the patterns of change depended on the cryogenic treatment. Our results provide new insights into the cryopreservation of, and the lipid metabolism in, algae. PMID:26731741

  14. The Regulation of Photosynthetic Structure and Function during Nitrogen Deprivation in Chlamydomonas reinhardtii1[OPEN

    PubMed Central

    Juergens, Matthew T.; Deshpande, Rahul R.; Lucker, Ben F.; Park, Jeong-Jin; Wang, Hongxia; Gargouri, Mahmoud; Holguin, F. Omar; Disbrow, Bradley; Schaub, Tanner; Skepper, Jeremy N.; Kramer, David M.; Gang, David R.; Hicks, Leslie M.; Shachar-Hill, Yair

    2015-01-01

    The accumulation of carbon storage compounds by many unicellular algae after nutrient deprivation occurs despite declines in their photosynthetic apparatus. To understand the regulation and roles of photosynthesis during this potentially bioenergetically valuable process, we analyzed photosynthetic structure and function after nitrogen deprivation in the model alga Chlamydomonas reinhardtii. Transcriptomic, proteomic, metabolite, and lipid profiling and microscopic time course data were combined with multiple measures of photosynthetic function. Levels of transcripts and proteins of photosystems I and II and most antenna genes fell with differing trajectories; thylakoid membrane lipid levels decreased, while their proportions remained similar and thylakoid membrane organization appeared to be preserved. Cellular chlorophyll (Chl) content decreased more than 2-fold within 24 h, and we conclude from transcript protein and 13C labeling rates that Chl synthesis was down-regulated both pre- and posttranslationally and that Chl levels fell because of a rapid cessation in synthesis and dilution by cellular growth rather than because of degradation. Photosynthetically driven oxygen production and the efficiency of photosystem II as well as P700+ reduction and electrochromic shift kinetics all decreased over the time course, without evidence of substantial energy overflow. The results also indicate that linear electron flow fell approximately 15% more than cyclic flow over the first 24 h. Comparing Calvin-Benson cycle transcript and enzyme levels with changes in photosynthetic 13CO2 incorporation rates also pointed to a coordinated multilevel down-regulation of photosynthetic fluxes during starch synthesis before the induction of high triacylglycerol accumulation rates. PMID:25489023

  15. Method to assemble and integrate biochemical pathways into the chloroplast genome of Chlamydomonas reinhardtii.

    PubMed

    Noor-Mohammadi, Samaneh; Pourmir, Azadeh; Johannes, Tyler W

    2012-11-01

    Recombinant protein expression in the chloroplasts of green algae has recently become more routine; however, the heterologous expression of multiple proteins or complete biosynthetic pathways remains a significant challenge. Here, we show that a modified DNA Assembler approach can be used to rapidly assemble multiple-gene biosynthetic pathways in yeast and then integrate these assembled pathways at a site-specific location in the chloroplast genome of the microalgal species Chlamydomonas reinhardtii. As a proof of concept, this method was used to successfully integrate and functionally express up to three reporter proteins (AphA6, AadA, and GFP) in the chloroplast of C. reinhardtii. An analysis of the relative gene expression of the engineered strains showed significant differences in the mRNA expression levels of the reporter genes and thus highlights the importance of proper promoter/untranslated region selection when constructing a target pathway. This new method represents a useful genetic tool in the construction and integration of complex biochemical pathways into the chloroplast genome of microalgae and should aid current efforts to engineer algae for biofuels production and other desirable natural products.

  16. High light induced changes in organization, protein profile and function of photosynthetic machinery in Chlamydomonas reinhardtii.

    PubMed

    Nama, Srilatha; Madireddi, Sai Kiran; Devadasu, Elsin Raju; Subramanyam, Rajagopal

    2015-11-01

    The green alga Chlamydomonas (C.) reinhardtii is used as a model organism to understand the efficiency of photosynthesis along with the organization and protein profile of photosynthetic apparatus under various intensities of high light exposure for 1h. Chlorophyll (Chl) a fluorescence induction, OJIPSMT transient was decreased with increase in light intensity indicating the reduction in photochemical efficiency. Further, circular dichroism studies of isolated thylakoids from high light exposed cells showed considerable change in the pigment-pigment interactions and pigment-proteins interactions. Furthermore, the organization of supercomplexes from thylakoids is studied, in which, one of the hetero-trimer of light harvesting complex (LHC) II is affected significantly in comparison to other complexes of LHC's monomers. Also, other supercomplexes, photosystem (PS)II reaction center dimer and PSI complexes are reduced. Additionally, immunoblot analysis of thylakoid proteins revealed that PSII core proteins D1 and D2 were significantly decreased during high light treatment. Similarly, the PSI core proteins PsaC, PsaD and PsaG were drastically changed. Further, the LHC antenna proteins of PSI and PSII were differentially affected. From our results it is clear that LHCs are damaged significantly, consequently the excitation energy is not efficiently transferred to the reaction center. Thus, the photochemical energy transfer from PSII to PSI is reduced. The inference of the study deciphers the structural and functional changes driven by light may therefore provide plants/alga to regulate the light harvesting capacity in excess light conditions.

  17. The microalga Chlamydomonas reinhardtii as a platform for the production of human protein therapeutics

    PubMed Central

    Rasala, Beth A

    2011-01-01

    Microalgae are a diverse group of eukaryotic photosynthetic microorganisms. While microalgae play a crucial role in global carbon fixation and oxygen evolution, these organisms have recently gained much attention for their potential role in biotechnological and industrial applications, such as the production of biofuels. We investigated the potential of the microalga Chlamydomonas reinhardtii to be a platform for the production of human therapeutic proteins. C. reinhardtii is a unicellular freshwater green alga that has served as a popular model alga for physiological, molecular, biochemical and genetic studies. As such, the molecular toolkit for this microorganism is highly developed, including well-established methods for genetic transformation and recombinant gene expression. We transformed the chloroplast genome of C. reinhardtii with seven unrelated genes encoding for current or potential human therapeutic proteins and found that four of these genes supported protein accumulation to levels that are sufficient for commercial production. Furthermore, the algal-produced proteins were bioactive. Thus, the microalga C. reinhardtii has the potential to be a robust platform for human therapeutic protein production. PMID:21636988

  18. The circadian clock of the unicellular eukaryotic model organism Chlamydomonas reinhardtii.

    PubMed

    Mittag, Maria; Wagner, Volker

    2003-05-01

    The green unicellular alga Chlamydomonas reinhardtii, also called 'green yeast', emerged in the past years as a model organism for specific scientific questions such as chloroplast biogenesis and function, the composition of the flagella including its basal apparatus, or the mechanism of the circadian clock. Sequencing of its chloroplast and mitochondrial genomes have already been completed and a first draft of its nuclear genome has also been released recently. In C. reinhardtii several circadian rhythms are physiologically well characterized, and one of them has even been shown to operate in outer space. Circadian expression patterns of nuclear and plastid genes have been studied. The mode of regulation of these genes occurs at the transcriptional level, although there is also evidence for posttranscriptional control. A clock-controlled, phylogenetically conserved RNA-binding protein was characterized in this alga, which interacts with several mRNAs that all contain a common cis-acting motif. Its function within the circadian system is currently under investigation. This review summarizes the current state of the knowledge about the circadian system in C. reinhardtii and points out its potential for future studies.

  19. Metabolic control of urea catabolism in Chlamydomonas reinhardi and Chlorella pyrenoidosa.

    PubMed Central

    Hodson, R C; Williams, S K; Davidson, W R

    1975-01-01

    In the unicellular green alga Chlamydomonas reinhardi (strain y-1), synthesis of the enzymes required for urea hydrolysis is under substrate induction control by urea and under end product repression control by ammonia. Hydrolysis of urea if effected by the sequential action of the discrete enzymes urea carboxylase and allophanate lyase, collectively called urea amidolyase. The carboxylase converts urea to allophanate in a reaction requiring biotin, adenosine 5'-triphosphate, and Mg2+. The lyase hydrolzyes allophanate to ammonium ions and bicarbonate. Neither activity is present in more than trace amounts when cultures are grown with ammonia or urea plus ammonia, or when they are starved for nitrogen for 8 h. Urea in the absence of ammonia induces both activities 10 to 100 times the basal levels. Addition of ammonia to an induced culture causes complete cessation of carboxylase accumulation and an 80% depression of lyase accumulation. Ammonia does not reduce urea uptake by repressed cells, so it does not prevent induction by the mechanism of inducer exclusion. The unicellular green alga Chlorella pyrenoidosa (strain 3 Emerson) also has discrete carboxylase and lyase enzymes, but only the carboxylase exhibits metabolic control. PMID:1116994

  20. Algae Derived Biofuel

    SciTech Connect

    Jahan, Kauser

    2015-03-31

    One of the most promising fuel alternatives is algae biodiesel. Algae reproduce quickly, produce oils more efficiently than crop plants, and require relatively few nutrients for growth. These nutrients can potentially be derived from inexpensive waste sources such as flue gas and wastewater, providing a mutual benefit of helping to mitigate carbon dioxide waste. Algae can also be grown on land unsuitable for agricultural purposes, eliminating competition with food sources. This project focused on cultivating select algae species under various environmental conditions to optimize oil yield. Membrane studies were also conducted to transfer carbon di-oxide more efficiently. An LCA study was also conducted to investigate the energy intensive steps in algae cultivation.

  1. Structure of GUN4 from Chlamydomonas reinhardtii.

    PubMed

    Tarahi Tabrizi, Shabnam; Langley, David B; Harrop, Stephen J; Duff, Anthony P; Willows, Robert D

    2015-08-01

    The genomes uncoupled 4 (GUN4) protein stimulates chlorophyll biosynthesis by increasing the activity of Mg-chelatase, the enzyme that inserts magnesium into protoporphyrin IX (PPIX) in the chlorophyll biosynthesis pathway. One of the roles of GUN4 is in binding PPIX and Mg-PPIX. In eukaryotes, GUN4 also participates in plastid-to-nucleus signalling, although the mechanism for this is unclear. Here, the first crystal structure of a eukaryotic GUN4, from Chlamydomonas reinhardtii, is presented. The structure is in broad agreement with those of previously solved cyanobacterial structures. Most interestingly, conformational divergence is restricted to several loops which cover the porphyrin-binding cleft. The conformational dynamics suggested by this ensemble of structures lend support to the understanding of how GUN4 binds PPIX or Mg-PPIX.

  2. Micro-algae come of age as a platform for recombinant protein production.

    PubMed

    Specht, Elizabeth; Miyake-Stoner, Shigeki; Mayfield, Stephen

    2010-10-01

    A complete set of genetic tools is still being developed for the micro-alga Chlamydomonas reinhardtii. Yet even with this incomplete set, this photosynthetic single-celled plant has demonstrated significant promise as a platform for recombinant protein expression. In recent years, techniques have been developed that allow for robust expression of genes from both the nuclear and plastid genome. With these advances, many research groups have examined the pliability of this and other micro-algae as biological machines capable of producing recombinant peptides and proteins. This review describes recent successes in recombinant protein production in Chlamydomonas, including production of complex mammalian therapeutic proteins and monoclonal antibodies at levels sufficient for production at economic parity with existing production platforms. These advances have also shed light on the details of algal protein production at the molecular level, and provide insight into the next steps for optimizing micro-algae as a useful platform for the production of therapeutic and industrially relevant recombinant proteins.

  3. Bioaccumulation and catabolism of prometryne in green algae.

    PubMed

    Jin, Zhen Peng; Luo, Kai; Zhang, Shuang; Zheng, Qi; Yang, Hong

    2012-04-01

    Investigation on organic xenobiotics bioaccumulation/biodegradation in green algae is of great importance from environmental point of view because widespread distribution of these compounds in agricultural areas has become one of the major problems in aquatic ecosystem. Also, new technology needs to be developed for environmental detection and re-usage of the compounds as bioresources. Prometryne as a herbicide is widely used for killing annual grasses in China and other developing countries. However, overuse of the pesticide results in high risks to contamination to aquatic environments. In this study, we focused on analysis of bioaccumulation and degradation of prometryne in Chlamydomonas reinhardtii, a green alga, along with its adaptive response to prometryne toxicity. C. reinhardtii treated with prometryne at 2.5-12.5 μg L(-1) for 4 d or 7.5 μg L(-1) for 1-6 d accumulated a large quantity of prometryne, with more than 2 mg kg(-1) fresh weight in cells exposed to 10 μg L(-1) prometryne. Moreover, it showed a great ability to degrade simultaneously the cell-accumulated prometryne. Such uptake and catabolism of prometryne led to the rapid removal of prometryne from media. Physiological and molecular analysis revealed that toxicology was associated with accumulation of prometryne in the cells. The biological processes of degradation can be interpreted as an internal tolerance mechanism. These results suggest that the green alga is useful in bioremediation of prometryne-contaminated aquatic ecosystems.

  4. An exogenous chloroplast genome for complex sequence manipulation in algae

    PubMed Central

    O'Neill, Bryan M.; Mikkelson, Kari L.; Gutierrez, Noel M.; Cunningham, Jennifer L.; Wolff, Kari L.; Szyjka, Shawn J.; Yohn, Christopher B.; Redding, Kevin E.; Mendez, Michael J.

    2012-01-01

    We demonstrate a system for cloning and modifying the chloroplast genome from the green alga, Chlamydomonas reinhardtii. Through extensive use of sequence stabilization strategies, the ex vivo genome is assembled in yeast from a collection of overlapping fragments. The assembled genome is then moved into bacteria for large-scale preparations and transformed into C. reinhardtii cells. This system also allows for the generation of simultaneous, systematic and complex genetic modifications at multiple loci in vivo. We use this system to substitute genes encoding core subunits of the photosynthetic apparatus with orthologs from a related alga, Scenedesmus obliquus. Once transformed into algae, the substituted genome recombines with the endogenous genome, resulting in a hybrid plastome comprising modifications in disparate loci. The in vivo function of the genomes described herein demonstrates that simultaneous engineering of multiple sites within the chloroplast genome is now possible. This work represents the first steps toward a novel approach for creating genetic diversity in any or all regions of a chloroplast genome. PMID:22116061

  5. Zinc Deficiency Impacts CO2 Assimilation and Disrupts Copper Homeostasis in Chlamydomonas reinhardtii*

    PubMed Central

    Malasarn, Davin; Kropat, Janette; Hsieh, Scott I.; Finazzi, Giovanni; Casero, David; Loo, Joseph A.; Pellegrini, Matteo; Wollman, Francis-André; Merchant, Sabeeha S.

    2013-01-01

    Zinc is an essential nutrient because of its role in catalysis and in protein stabilization, but excess zinc is deleterious. We distinguished four nutritional zinc states in the alga Chlamydomonas reinhardtii: toxic, replete, deficient, and limited. Growth is inhibited in zinc-limited and zinc-toxic cells relative to zinc-replete cells, whereas zinc deficiency is visually asymptomatic but distinguished by the accumulation of transcripts encoding ZIP family transporters. To identify targets of zinc deficiency and mechanisms of zinc acclimation, we used RNA-seq to probe zinc nutrition-responsive changes in gene expression. We identified genes encoding zinc-handling components, including ZIP family transporters and candidate chaperones. Additionally, we noted an impact on two other regulatory pathways, the carbon-concentrating mechanism (CCM) and the nutritional copper regulon. Targets of transcription factor Ccm1 and various CAH genes are up-regulated in zinc deficiency, probably due to reduced carbonic anhydrase activity, validated by quantitative proteomics and immunoblot analysis of Cah1, Cah3, and Cah4. Chlamydomonas is therefore not able to grow photoautotrophically in zinc-limiting conditions, but supplementation with 1% CO2 restores growth to wild-type rates, suggesting that the inability to maintain CCM is a major consequence of zinc limitation. The Crr1 regulon responds to copper limitation and is turned on in zinc deficiency, and Crr1 is required for growth in zinc-limiting conditions. Zinc-deficient cells are functionally copper-deficient, although they hyperaccumulate copper up to 50-fold over normal levels. We suggest that zinc-deficient cells sequester copper in a biounavailable form, perhaps to prevent mismetallation of critical zinc sites. PMID:23439652

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

    PubMed Central

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

    2011-01-01

    Summary 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. PMID:21309872

  7. How Chlamydomonas keeps track of the light once it has reached the right phototactic orientation.

    PubMed Central

    Schaller, K; David, R; Uhl, R

    1997-01-01

    By using a real-time assay that allows measurement of the phototactic orientation of the unicellular alga Chlamydomonas with millisecond time resolution, it can be shown that single photons not only induce transient direction changes but that fluence rates as low as 1 photon cell(-1) s(-1) can already lead to a persistent orientation. Orientation is a binary variable, i.e., in a partially oriented population some organisms are fully oriented while the rest are still at random. Action spectra reveal that the response to a pulsed stimulus follows the Dartnall-nomogram for a rhodopsin while the response to a persistent stimulus falls off more rapidly toward the red end of the spectrum. Thus light of 540 nm, for which chlamy-rhodopsin is equally sensitive as for 440-nm light, induces no measurable persistent orientation while 440-nm light does. A model is presented which explains not only this behavior, but also how Chlamydomonas can track the light direction and switches between a positive and negative phototaxis. According to the model the ability to detect the direction of light, to make the right turn and to stay oriented, is a direct consequence of the helical path of the organism, the orientation of its eyespot relative to the helix-axis, and the special shielding properties of eyespot and cell body. The model places particular emphasis on the fact that prolonged swimming into the correct direction not only requires making a correct turn initially, but also avoiding further turns once the right direction has been reached. Images FIGURE 1 FIGURE 4 FIGURE 6 FIGURE 7 FIGURE 8 PMID:9284323

  8. Analytical approaches to photobiological hydrogen production in unicellular green algae.

    PubMed

    Hemschemeier, Anja; Melis, Anastasios; Happe, Thomas

    2009-01-01

    Several species of unicellular green algae, such as the model green microalga Chlamydomonas reinhardtii, can operate under either aerobic photosynthesis or anaerobic metabolism conditions. A particularly interesting metabolic condition is that of "anaerobic oxygenic photosynthesis", whereby photosynthetically generated oxygen is consumed by the cell's own respiration, causing anaerobiosis in the culture in the light, and induction of the cellular "hydrogen metabolism" process. The latter entails an alternative photosynthetic electron transport pathway, through the oxygen-sensitive FeFe-hydrogenase, leading to the light-dependent generation of molecular hydrogen in the chloroplast. The FeFe-hydrogenase is coupled to the reducing site of photosystem-I via ferredoxin and is employed as an electron-pressure valve, through which electrons are dissipated, thus permitting a sustained electron transport in the thylakoid membrane of photosynthesis. This hydrogen gas generating process in the cells offers testimony to the unique photosynthetic metabolism that can be found in many species of green microalgae. Moreover, it has attracted interest by the biotechnology and bioenergy sectors, as it promises utilization of green microalgae and the process of photosynthesis in renewable energy production. This article provides an overview of the principles of photobiological hydrogen production in microalgae and addresses in detail the process of induction and analysis of the hydrogen metabolism in the cells. Furthermore, methods are discussed by which the interaction of photosynthesis, respiration, cellular metabolism, and H(2) production in Chlamydomonas can be monitored and regulated.

  9. CSL encodes a leucine-rich-repeat protein implicated in red/violet light signaling to the circadian clock in Chlamydomonas.

    PubMed

    Kinoshita, Ayumi; Niwa, Yoshimi; Onai, Kiyoshi; Yamano, Takashi; Fukuzawa, Hideya; Ishiura, Masahiro; Matsuo, Takuya

    2017-03-01

    The green alga Chlamydomonas reinhardtii shows various light responses in behavior and physiology. One such photoresponse is the circadian clock, which can be reset by external light signals to entrain its oscillation to daily environmental cycles. In a previous report, we suggested that a light-induced degradation of the clock protein ROC15 is a trigger to reset the circadian clock in Chlamydomonas. However, light signaling pathways of this process remained unclear. Here, we screened for mutants that show abnormal ROC15 diurnal rhythms, including the light-induced protein degradation at dawn, using a luciferase fusion reporter. In one mutant, ROC15 degradation and phase resetting of the circadian clock by light were impaired. Interestingly, the impairments were observed in response to red and violet light, but not to blue light. We revealed that an uncharacterized gene encoding a protein similar to RAS-signaling-related leucine-rich repeat (LRR) proteins is responsible for the mutant phenotypes. Our results indicate that a previously uncharacterized red/violet light signaling pathway is involved in the phase resetting of circadian clock in Chlamydomonas.

  10. Proteomic profiling of oil bodies isolated from the unicellular green microalga Chlamydomonas reinhardtii: with focus on proteins involved in lipid metabolism.

    PubMed

    Nguyen, Hoa M; Baudet, Mathieu; Cuiné, Stéphan; Adriano, Jean-Marc; Barthe, Damien; Billon, Emmanuelle; Bruley, Christophe; Beisson, Fred; Peltier, Gilles; Ferro, Myriam; Li-Beisson, Yonghua

    2011-11-01

    Oil bodies are sites of energy and carbon storage in many organisms including microalgae. As a step toward deciphering oil accumulation mechanisms in algae, we used proteomics to analyze purified oil bodies from the model microalga Chlamydomonas reinhardtii grown under nitrogen deprivation. Among the 248 proteins (≥ 2 peptides) identified by LC-MS/MS, 33 were putatively involved in the metabolism of lipids (mostly acyl-lipids and sterols). Compared with a recently reported Chlamydomonas oil body proteome, 19 new proteins of lipid metabolism were identified, spanning the key steps of the triacylglycerol synthesis pathway and including a glycerol-3-phosphate acyltransferase (GPAT), a lysophosphatidic acid acyltransferase (LPAT) and a putative phospholipid:diacylglycerol acyltransferase (PDAT). In addition, proteins putatively involved in deacylation/reacylation, sterol synthesis, lipid signaling and lipid trafficking were found to be associated with the oil body fraction. This data set thus provides evidence that Chlamydomonas oil bodies are not only storage compartments but also are dynamic structures likely to be involved in processes such as oil synthesis, degradation and lipid homeostasis. The proteins identified here should provide useful targets for genetic studies aiming at increasing our understanding of triacyglycerol synthesis and the role of oil bodies in microalgal cell functions.

  11. CSL encodes a leucine-rich-repeat protein implicated in red/violet light signaling to the circadian clock in Chlamydomonas

    PubMed Central

    Kinoshita, Ayumi; Niwa, Yoshimi; Onai, Kiyoshi; Fukuzawa, Hideya; Ishiura, Masahiro

    2017-01-01

    The green alga Chlamydomonas reinhardtii shows various light responses in behavior and physiology. One such photoresponse is the circadian clock, which can be reset by external light signals to entrain its oscillation to daily environmental cycles. In a previous report, we suggested that a light-induced degradation of the clock protein ROC15 is a trigger to reset the circadian clock in Chlamydomonas. However, light signaling pathways of this process remained unclear. Here, we screened for mutants that show abnormal ROC15 diurnal rhythms, including the light-induced protein degradation at dawn, using a luciferase fusion reporter. In one mutant, ROC15 degradation and phase resetting of the circadian clock by light were impaired. Interestingly, the impairments were observed in response to red and violet light, but not to blue light. We revealed that an uncharacterized gene encoding a protein similar to RAS-signaling-related leucine-rich repeat (LRR) proteins is responsible for the mutant phenotypes. Our results indicate that a previously uncharacterized red/violet light signaling pathway is involved in the phase resetting of circadian clock in Chlamydomonas. PMID:28333924

  12. The small domain of cytochrome f from the psychrophile Chlamydomonas raudensis UWO 241 modulates the apparent molecular mass and decreases the accumulation of cytochrome f in the mesophile Chlamydomonas reinhardtii.

    PubMed

    Gudynaite-Savitch, Loreta; Loiselay, Christelle; Savitch, Leonid V; Simmonds, John; Kohalmi, Susanne E; Choquet, Yves; Hüner, Norman P A

    2007-10-01

    Cytochrome f from the psychrophile Chlamydomonas raudensis UWO 241 has a lower thermostability of its c-type heme and an apparent molecular mass that is 7 kDa lower than that of the model mesophilic green alga Chlamydomonas reinhardtii. We combined chloroplast transformation, site-directed mutagensis, and the creation of chimeric fusion constructs to assess the contribution of specific domains and (or) amino acids residues to the structure, stability, and accumulation of cytochrome f, as well as its function in photosynthetic intersystem electron transport. We demonstrate that differences in the amino acid sequence of the small domain and specific charged amino acids in the large domain of cytochrome f alter the physical properties of this protein but do not affect either the thermostability of the c-type heme, the apparent half-life of cytochrome f in the presence of the chloroplastic protein synthesis inhibitor chloramphenicol, or the capacity for photosynthetic intersystem electron transport, measured as e-/P700. However, pulse-labeling with [14C]acetate, combined with immunoblotting, indicated that the negative autoregulation of cytochrome f accumulation observed in mesophilic C. reinhardtii transformed with chimeric constructs from the psychrophile was likely the result of the defective association of the chimeric forms of cytochrome f with the other subunits of the cytochrome b6/f complex native to the C. reinhardtii wild type. These results are discussed in terms of the unique fatty acid composition of the thylakoid membranes of C. raudensis UWO 241 adapted to cold environments.

  13. Enhanced genetic tools for engineering multigene traits into green algae.

    PubMed

    Rasala, Beth A; Chao, Syh-Shiuan; Pier, Matthew; Barrera, Daniel J; Mayfield, Stephen P

    2014-01-01

    Transgenic microalgae have the potential to impact many diverse biotechnological industries including energy, human and animal nutrition, pharmaceuticals, health and beauty, and specialty chemicals. However, major obstacles to sophisticated genetic and metabolic engineering in algae have been the lack of well-characterized transformation vectors to direct engineered gene products to specific subcellular locations, and the inability to robustly express multiple nuclear-encoded transgenes within a single cell. Here we validate a set of genetic tools that enable protein targeting to distinct subcellular locations, and present two complementary methods for multigene engineering in the eukaryotic green microalga Chlamydomonas reinhardtii. The tools described here will enable advanced metabolic and genetic engineering to promote microalgae biotechnology and product commercialization.

  14. Amidic and acetonic cryoprotectants improve cryopreservation of volvocine green algae.

    PubMed

    Nakazawa, A; Nishii, I

    2012-01-01

    A number of volvocalean green algae species were subjected to a two-step cryopreservation protocol with various cryoprotectants. Potential cryoprotectants were methanol (DMSO), N,N-dimethylformamide (DMF), N,N-dimethylacetamide, N-methylformamide, and hydroxyacetone (HA). We confirmed prior reports that MeOH was effective for cryopreserving Chlamydomonas, but did not work well for larger volvocaleans such as Volvox. In contrast, DMF and HA were effective for both unicellular and multicellular representatives. When we used a cold-inducible transposon to probe Southern blots of Volvox DNA samples taken before and after storage for one month in LN, we could detect no differences, indicating that the genome had remained relatively stable and that the transposon had not been induced by the cryopreservation procedure. We believe these methods will facilitate long-term storage of several volvocine algal species, including Volvox strains harboring transposon-induced mutations of developmental interest.

  15. Surface gas-exchange processes of snow algae

    PubMed Central

    Williams, William E.; Gorton, Holly L.; Vogelmann, Thomas C.

    2003-01-01

    The red-colored chlorophyte Chlamydomonas nivalis is commonly found in summer snowfields. We used a modified Li-Cor gas-exchange system to investigate surface gas-exchange characteristics of snow colonized by this alga, finding rates of CO2 uptake up to 0.3 μmol m−2⋅s−1 in dense algal blooms. Experiments varying the irradiance resulted in light curves that resembled those of the leaves of higher plants. Red light was more effective than white and much more effective than green or blue, because of the red astaxanthin that surrounds and masks the algal chloroplasts. Integrating daily course measurements of gas exchange showed CO2 uptake around 2,300 μmol⋅m−2⋅day−1 in heavily colonized patches, indicating that summer snowfields can be surprisingly productive. PMID:12518048

  16. [Bacterial communities of brown and red algae from Peter the Great Bay, the Sea of Japan].

    PubMed

    Beleneva, I A; Zhukova, N V

    2006-01-01

    The structure of microbial communities of brown algae, red algae, and of the red alga Gracilaria verrucosa, healthy and affected with rotten thallus, were comparatively investigated; 61 strains of heterotrophic bacteria were isolated and characterized. Most of them were identified to the genus level, some Vibrio spp., to the species level according to their phenotypic properties and the fatty acid composition of cellular lipids. The composition of the microflora of two species of brown algae was different. In Chordaria flagelliformis, Pseudomonas spp. prevailed, and in Desmarestia viridis, Bacillus spp. The composition of the microflora of two red algae, G. verrucosa and Camphylaephora hyphaeoides, differed mainly in the ratio of prevailing groups of bacteria. The most abundant were bacteria of the CFB cluster and pseudoalteromonads. In addition, the following bacteria were found on the surface of the algae: Sulfitobacter spp., Halomonas spp., Acinetobacter sp., Planococcus sp., Arthrobacter sp., and Agromyces sp. From tissues of the affected G. verrucosa, only vibrios were isolated, both agarolytic and nonagarolytic. The existence of specific bacterial communities characteristic of different species of algae is suggested and the relation of Vibrio sp. to the pathological process in the tissues of G. verrucosa is supposed.

  17. Multiple stressor effects of predation by rotifers and herbicide pollution on different Chlamydomonas strains and potential impacts on population dynamics.

    PubMed

    Fischer, Beat B; Roffler, Severin; Eggen, Rik I L

    2012-12-01

    Environmental factors can interact with the effects of chemical pollutants on natural systems by inducing multiple stressor effects in individual organisms as well as by altering selection pressure on tolerant strains in heterogeneous populations. Predation is a stressful environmental factor relevant for many species. Therefore, the impact of predation by the rotifer Brachionus calyciflorus on tolerance of eight genetically different strains of the green alga Chlamydomonas reinhardtii to simultaneous exposure to each of the three herbicides (diuron, paraquat, and S-metolachlor) was tested. Interactions of combined stressors were analyzed based on the independent action model; additive, synergistic, and antagonistic effects of the combined exposure could be detected depending on the herbicide and strain tested. If cultures were acclimated (pre-exposed) to one stressor, tolerance to the second stressor could be increased. This indicates that physiological changes can induce cotolerance of predation-exposed algae to herbicides and of herbicide-treated algae to predation depending on the combination of stressors. The strain-specific differences in multiple stressor effects also changed the correlation of strains' tolerances to individual stressors determined during combined and single-stressor exposure. Changes in cotolerance to stressors affect selection pressure and population dynamics during long-term exposure. This shows that predation stress can have adverse effects on the toxicity of chemical pollutants to microalgae on the organism and population levels.

  18. Control of Autophagy in Chlamydomonas Is Mediated through Redox-Dependent Inactivation of the ATG4 Protease.

    PubMed

    Pérez-Pérez, María Esther; Lemaire, Stéphane D; Crespo, José L

    2016-12-01

    Autophagy is a major catabolic pathway by which eukaryotic cells deliver unnecessary or damaged cytoplasmic material to the vacuole for its degradation and recycling in order to maintain cellular homeostasis. Control of autophagy has been associated with the production of reactive oxygen species in several organisms, including plants and algae, but the precise regulatory molecular mechanisms remain unclear. Here, we show that the ATG4 protease, an essential protein for autophagosome biogenesis, plays a central role for the redox regulation of autophagy in the model green alga Chlamydomonas reinhardtii Our results indicate that the activity of C. reinhardtii ATG4 is regulated by the formation of a single disulfide bond with a low redox potential that can be efficiently reduced by the NADPH/thioredoxin system. Moreover, we found that treatment of C. reinhardtii cells with norflurazon, an inhibitor of carotenoid biosynthesis that generates reactive oxygen species and triggers autophagy in this alga, promotes the oxidation and aggregation of ATG4. We propose that the activity of the ATG4 protease is finely regulated by the intracellular redox state, and it is inhibited under stress conditions to ensure lipidation of ATG8 and thus autophagy progression in C. reinhardtii.

  19. Three Acyltransferases and Nitrogen-responsive Regulator Are Implicated in Nitrogen Starvation-induced Triacylglycerol Accumulation in Chlamydomonas*

    PubMed Central

    Boyle, Nanette R.; Page, Mark Dudley; Liu, Bensheng; Blaby, Ian K.; Casero, David; Kropat, Janette; Cokus, Shawn J.; Hong-Hermesdorf, Anne; Shaw, Johnathan; Karpowicz, Steven J.; Gallaher, Sean D.; Johnson, Shannon; Benning, Christoph; Pellegrini, Matteo; Grossman, Arthur; Merchant, Sabeeha S.

    2012-01-01

    Algae have recently gained attention as a potential source for biodiesel; however, much is still unknown about the biological triggers that cause the production of triacylglycerols. We used RNA-Seq as a tool for discovering genes responsible for triacylglycerol (TAG) production in Chlamydomonas and for the regulatory components that activate the pathway. Three genes encoding acyltransferases, DGAT1, DGTT1, and PDAT1, are induced by nitrogen starvation and are likely to have a role in TAG accumulation based on their patterns of expression. DGAT1 and DGTT1 also show increased mRNA abundance in other TAG-accumulating conditions (minus sulfur, minus phosphorus, minus zinc, and minus iron). Insertional mutants, pdat1-1 and pdat1-2, accumulate 25% less TAG compared with the parent strain, CC-4425, which demonstrates the relevance of the trans-acylation pathway in Chlamydomonas. The biochemical functions of DGTT1 and PDAT1 were validated by rescue of oleic acid sensitivity and restoration of TAG accumulation in a yeast strain lacking all acyltransferase activity. Time course analyses suggest than a SQUAMOSA promoter-binding protein domain transcription factor, whose mRNA increases precede that of lipid biosynthesis genes like DGAT1, is a candidate regulator of the nitrogen deficiency responses. An insertional mutant, nrr1-1, accumulates only 50% of the TAG compared with the parental strain in nitrogen-starvation conditions and is unaffected by other nutrient stresses, suggesting the specificity of this regulator for nitrogen-deprivation conditions. PMID:22403401

  20. Efficient Heterologous Transformation of Chlamydomonas reinhardtii npq2 Mutant with the Zeaxanthin Epoxidase Gene Isolated and Characterized from Chlorella zofingiensis

    PubMed Central

    Couso, Inmaculada; Cordero, Baldo F.; Vargas, María Ángeles; Rodríguez, Herminia

    2012-01-01

    In the violaxanthin cycle, the violaxanthin de-epoxidase and zeaxanthin epoxidase catalyze the inter-conversion between violaxanthin and zeaxanthin in both plants and green algae. The zeaxanthin epoxidase gene from the green microalga Chlorella zofingiensis (Czzep) has been isolated. This gene encodes a polypeptide of 596 amino acids. A single copy of Czzep has been found in the C. zofingiensis genome by Southern blot analysis. qPCR analysis has shown that transcript levels of Czzep were increased after zeaxanthin formation under high light conditions. The functionality of Czzep gene by heterologous genetic complementation in the Chlamydomonas mutant npq2, which lacks zeaxanthin epoxidase (ZEP) activity and accumulates zeaxanthin in all conditions, was analyzed. The Czzep gene was adequately inserted in the pSI105 vector and expressed in npq2. The positive transformants were able to efficiently convert zeaxanthin into violaxanthin, as well as to restore their maximum quantum efficiency of the PSII (Fv/Fm). These results show that Chlamydomonas can be an efficient tool for heterologous expression and metabolic engineering for biotechnological applications. PMID:23118714

  1. An Indexed, Mapped Mutant Library Enables Reverse Genetics Studies of Biological Processes in Chlamydomonas reinhardtii[OPEN

    PubMed Central

    Gang, Spencer S.; Blum, Sean R.; Ivanova, Nina; Yue, Rebecca; Grossman, Arthur R.

    2016-01-01

    The green alga Chlamydomonas reinhardtii is a leading unicellular model for dissecting biological processes in photosynthetic eukaryotes. However, its usefulness has been limited by difficulties in obtaining mutants in specific genes of interest. To allow generation of large numbers of mapped mutants, we developed high-throughput methods that (1) enable easy maintenance of tens of thousands of Chlamydomonas strains by propagation on agar media and by cryogenic storage, (2) identify mutagenic insertion sites and physical coordinates in these collections, and (3) validate the insertion sites in pools of mutants by obtaining >500 bp of flanking genomic sequences. We used these approaches to construct a stably maintained library of 1935 mapped mutants, representing disruptions in 1562 genes. We further characterized randomly selected mutants and found that 33 out of 44 insertion sites (75%) could be confirmed by PCR, and 17 out of 23 mutants (74%) contained a single insertion. To demonstrate the power of this library for elucidating biological processes, we analyzed the lipid content of mutants disrupted in genes encoding proteins of the algal lipid droplet proteome. This study revealed a central role of the long-chain acyl-CoA synthetase LCS2 in the production of triacylglycerol from de novo-synthesized fatty acids. PMID:26764374

  2. Species-specific isotope tracers to study the accumulation and biotransformation of mixtures of inorganic and methyl mercury by the microalga Chlamydomonas reinhardtii.

    PubMed

    Bravo, Andrea Garcia; Le Faucheur, Séverine; Monperrus, Mathilde; Amouroux, David; Slaveykova, Vera I

    2014-09-01

    The present study demonstrates that species-specific isotope tracing is an useful tool to precisely measure Hg accumulation and transformations capabilities of living organisms at concentrations naturally encountered in the environment. To that end, a phytoplanktonic green alga Chlamydomonas reinhardtii Dangeard (Chlamydomonadales, Chlorophyceae) was exposed to mixtures of (199)-isotopically enriched inorganic mercury ((199)IHg) and of (201)-isotopically enriched monomethylmercury ((201)CH3Hg) at a concentration range between less than 1 pM to 4 nM. Additionally, one exposure concentration of both mercury species was also studied separately to evaluate possible interactive effects. No difference in the intracellular contents was observed for algae exposed to (199)IHg and (201)CH3Hg alone or in their mixture, suggesting similar accumulation capacity for both species at the studied concentrations. Demethylation of (201)CH3Hg was observed at the highest exposure concentrations, whereas no methylation was detected.

  3. A new class of cyclin dependent kinase in Chlamydomonas is required for coupling cell size to cell division.

    PubMed

    Li, Yubing; Liu, Dianyi; López-Paz, Cristina; Olson, Bradley Jsc; Umen, James G

    2016-03-25

    Proliferating cells actively control their size by mechanisms that are poorly understood. The unicellular green alga Chlamydomonas reinhardtii divides by multiple fission, wherein a 'counting' mechanism couples mother cell-size to cell division number allowing production of uniform-sized daughters. We identified a sizer protein, CDKG1, that acts through the retinoblastoma (RB) tumor suppressor pathway as a D-cyclin-dependent RB kinase to regulate mitotic counting. Loss of CDKG1 leads to fewer mitotic divisions and large daughters, while mis-expression of CDKG1 causes supernumerous mitotic divisions and small daughters. The concentration of nuclear-localized CDKG1 in pre-mitotic cells is set by mother cell size, and its progressive dilution and degradation with each round of cell division may provide a link between mother cell-size and mitotic division number. Cell-size-dependent accumulation of limiting cell cycle regulators such as CDKG1 is a potentially general mechanism for size control.

  4. Chlamydomonas shortens its flagella by activating axonemal disassembly, stimulating IFT particle trafficking, and blocking anterograde cargo loading.

    PubMed

    Pan, Junmin; Snell, William J

    2005-09-01

    Almost all eukaryotic cells form cilia/flagella, maintain them at their genetically specified lengths, and shorten them. Here, we define the cellular mechanisms that bring about shortening of flagella prior to meiotic cell division and in response to environmental cues in the biflagellated green alga Chlamydomonas. We show that the flagellar shortening pathway is distinct from the one that enforces transient shortening essential for length control. During flagellar shortening, disassembly of the axoneme is stimulated over the basal rate, and the rate of entry into flagella of intraflagellar transport (IFT) particles is increased. Moreover, the particles entering the disassembling flagella lack cargo. Thus, flagellar shortening depends on the interplay between dynamic properties of the axoneme and the IFT machinery; a cell triggered to shorten its flagellum activates disassembly of the axoneme and stimulates entry into the flagellum of IFT particles possessing empty cargo binding sites available to retrieve the disassembled components.

  5. A sex recognition glycoprotein is encoded by the plus mating-type gene fus1 of Chlamydomonas reinhardtii.

    PubMed Central

    Ferris, P J; Woessner, J P; Goodenough, U W

    1996-01-01

    Sexual fusion between plus and minus gametes of the unicellular green alga Chlamydomonas reinhardtii entails adhesion between plus-specific and minus-specific "fringe" proteins displayed on the plasma membrane of gametic mating structures. We report the identification of the gene (fus1) encoding the plus fringe glycoprotein, which resides in a unique domain of the mating-type plus (mt+) locus, and which was identified by transposon insertions in three fusion-defective mutant strains. Transformation with fus1+ restores fringe and fusion competence to these mutants and to the pseudo-plus mutant imp11 mt-, defective in minus differentiation. The fus1 gene is remarkable in lacking the codon bias found in all other nuclear genes of C. reinhardtii. Images PMID:8856667

  6. Study of the effect of reducing conditions on the initial chlorophyll fluorescence rise in the green microalgae Chlamydomonas reinhardtii.

    PubMed

    Antal, T K; Kolacheva, A; Maslakov, A; Riznichenko, G Yu; Krendeleva, T E; Rubin, A B

    2013-03-01

    Incubation of Chlamydomonas reinhardtii cells under nutrient deficiency results in the faster initial rise in the light-induced chlorophyll fluorescence kinetic curve. We showed that short-term anaerobic incubation of algal cells altered initial fluorescence in a way similar to nutrient starvation, suggesting an important role of the plastoquinones redox state in the observed effect. Bi-component analysis of highly resolved initial fluorescence rise kinetics in sulfur- or oxygen-depleted C. reinhardtii cells suggested that one of the mechanisms underlying the observed phenomenon involves primary closure (photochemical inactivation via Qa reduction) of β-type PSII as compared to α-PSII. Moreover, results of modeling of the fluorescence curve brought us to the conclusion that accumulation of closed centers in α-PSII supercomplexes may also cause a faster initial fluorescence rise. The observed correlations between nutrient supply rate and initial fluorescence rise pattern in green algae can serve to characterize culture nutritional status in vivo.

  7. Repression of Essential Chloroplast Genes Reveals New Signaling Pathways and Regulatory Feedback Loops in Chlamydomonas[W

    PubMed Central

    Ramundo, Silvia; Rahire, Michèle; Schaad, Olivier; Rochaix, Jean-David

    2013-01-01

    Although reverse genetics has been used to elucidate the function of numerous chloroplast proteins, the characterization of essential plastid genes and their role in chloroplast biogenesis and cell survival has not yet been achieved. Therefore, we developed a robust repressible chloroplast gene expression system in the unicellular alga Chlamydomonas reinhardtii based mainly on a vitamin-repressible riboswitch, and we used this system to study the role of two essential chloroplast genes: ribosomal protein S12 (rps12), encoding a plastid ribosomal protein, and rpoA, encoding the α-subunit of chloroplast bacterial-like RNA polymerase. Repression of either of these two genes leads to the arrest of cell growth, and it induces a response that involves changes in expression of nuclear genes implicated in chloroplast biogenesis, protein turnover, and stress. This response also leads to the overaccumulation of several plastid transcripts and reveals the existence of multiple negative regulatory feedback loops in the chloroplast gene circuitry. PMID:23292734

  8. Retinal Chromophore Structure and Schiff Base Interactions in Red-Shifted Channelrhodopsin-1 from Chlamydomonas augustae

    PubMed Central

    2015-01-01

    Channelrhodopsins (ChRs), which form a distinct branch of the microbial rhodopsin family, control phototaxis in green algae. Because ChRs can be expressed and function in neuronal membranes as light-gated cation channels, they have rapidly become an important optogenetic tool in neurobiology. While channelrhodopsin-2 from the unicellular alga Chlamydomonas reinhardtii (CrChR2) is the most commonly used and extensively studied optogenetic ChR, little is known about the properties of the diverse group of other ChRs. In this study, near-infrared confocal resonance Raman spectroscopy along with hydrogen–deuterium exchange and site-directed mutagenesis were used to study the structure of red-shifted ChR1 from Chlamydomonas augustae (CaChR1). These measurements reveal that (i) CaChR1 has an all-trans-retinal structure similar to those of the light-driven proton pump bacteriorhodopsin (BR) and sensory rhodopsin II but different from that of the mixed retinal composition of CrChR2, (ii) lowering the pH from 7 to 2 or substituting neutral residues for Glu169 or Asp299 does not significantly shift the ethylenic stretch frequency more than 1–2 cm–1 in contrast to BR in which a downshift of 7–9 cm–1 occurs reflecting neutralization of the Asp85 counterion, and (iii) the CaChR1 protonated Schiff base (SB) has stronger hydrogen bonding than BR. A model is proposed to explain these results whereby at pH 7 the predominant counterion to the SB is Asp299 (the homologue to Asp212 in BR) while Glu169 (the homologue to Asp85 in BR) exists in a neutral state. We observe an unusual constancy of the resonance Raman spectra over the broad range from pH 9 to 2 and discuss its implications. These results are in accord with recent visible absorption and current measurements of CaChR1 [Sineshchekov, O. A., et al. (2013) Intramolecular proton transfer in channelrhodopsins. Biophys. J. 104, 807–817; Li, H., et al. (2014) Role of a helix B lysine residue in the photoactive site in

  9. A fluorescence-activated cell sorting-based strategy for rapid isolation of high-lipid Chlamydomonas mutants.

    PubMed

    Terashima, Mia; Freeman, Elizabeth S; Jinkerson, Robert E; Jonikas, Martin C

    2015-01-01

    There is significant interest in farming algae for the direct production of biofuels and valuable lipids. Chlamydomonas reinhardtii is the leading model system for studying lipid metabolism in green algae, but current methods for isolating mutants of this organism with a perturbed lipid content are slow and tedious. Here, we present the Chlamydomonas high-lipid sorting (CHiLiS) strategy, which enables enrichment of high-lipid mutants by fluorescence-activated cell sorting (FACS) of pooled mutants stained with the lipid-sensitive dye Nile Red. This method only takes 5 weeks from mutagenesis to mutant isolation. We developed a staining protocol that allows quantification of lipid content while preserving cell viability. We improved separation of high-lipid mutants from the wild type by using each cell's chlorophyll fluorescence as an internal control. We initially demonstrated 20-fold enrichment of the known high-lipid mutant sta1 from a mixture of sta1 and wild-type cells. We then applied CHiLiS to sort thousands of high-lipid cells from a pool of about 60,000 mutants. Flow cytometry analysis of 24 individual mutants isolated by this approach revealed that about 50% showed a reproducible high-lipid phenotype. We further characterized nine of the mutants with the highest lipid content by flame ionization detection and mass spectrometry lipidomics. All mutants analyzed had a higher triacylglycerol content and perturbed whole-cell fatty acid composition. One arbitrarily chosen mutant was evaluated by microscopy, revealing larger lipid droplets than the wild type. The unprecedented throughput of CHiLiS opens the door to a systems-level understanding of green algal lipid biology by enabling genome-saturating isolation of mutants in key genes.

  10. Patching Holes in the Chlamydomonas Genome

    PubMed Central

    Tulin, Frej; Cross, Frederick R.

    2016-01-01

    The Chlamydomonas genome has been sequenced, assembled, and annotated to produce a rich resource for genetics and molecular biology in this well-studied model organism. However, the current reference genome contains ∼1000 blocks of unknown sequence (‘N-islands’), which are frequently placed in introns of annotated gene models. We developed a strategy to search for previously unknown exons hidden within such blocks, and determine the sequence, and exon/intron boundaries, of such exons. These methods are based on assembly and alignment of short cDNA and genomic DNA reads, completely independent of prior reference assembly or annotation. Our evidence indicates that a substantial proportion of the annotated intronic N-islands contain hidden exons. For most of these, our algorithm recovers full exonic sequence with associated splice junctions and exon-adjacent intronic sequence. These new exons represent de novo sequence generally present nowhere in the assembled genome, and the added sequence improves evolutionary conservation of the predicted encoded peptides. PMID:27175017

  11. Recording and analyzing IFT in Chlamydomonas flagella.

    PubMed

    Dentler, William; Vanderwaal, Kristyn; Porter, Mary E

    2009-01-01

    The transport of materials to and from the cell body and tips of eukaryotic flagella and cilia is carried out by a process called intraflagellar transport, or IFT. This process is essential for the assembly and maintenance of cilia and flagella: in the absence of IFT, cilia cannot assemble and, if IFT is arrested in ciliated cells, the cilia disassemble. The major IFT complex proteins and the major motor proteins, kinesin-2 and osm-3 (which transport particles from the cell body to ciliary tips) and cytoplasmic dynein 1b (which transports particles from ciliary tips to the cell body) have been identified. However, we have little understanding of the structure of the IFT particles, the cargo that these particles carry, how cargo is loaded and unloaded from the particles, or how the motor proteins are regulated. The focus of this chapter is to provide methods to observe and quantify the movements of IFT particles in Chlamydomonas flagella. IFT movements can be visualized in paralyzed or partially arrested flagella using either differential interference contrast (IFT) microscopy or, in cells with fluorescently tagged IFT components, with fluorescence microscopy. Methods for recording IFT movements and analyzing movements using kymograms are described.

  12. Studies on flagellar shortening in Chlamydomonas reinhardtii

    SciTech Connect

    Cherniack, J.

    1985-01-01

    Flagellar shortening of Chlamydomonas reinhardtii was promoted by sodium chloride, pyrophosphate (sodium, potassium and ammonium salts), EDTA and EGTA, succinate, citrate and oxalate (sodium salts), caffeine and aminophylline. Removal of calcium from the medium potentiated the effects of these agents in inducing shortening. Investigations of the release of phosphorylated compounds to the medium during pyrophosphate-induced flagellar shortening of cells pre-labelled with /sup 32/P, revealed an as yet unidentified /sup 32/P-labelled compound with distinct chromatographic properties. Chromatography and electrophoresis indicates that it is a small, highly polar molecule with a high charge to mass ratio, containing thermo- and acid-labile phosphate linkages. Investigations showed of the release of /sup 35/S-labelled protein to the medium from cells pre-labelled with /sup 35/S-sulfate showed that flagellated cells released two prominent polypeptides which comigrated with ..cap alpha..- and ..beta..-flagellar tubulin on SDS polyacrylamide gel electrophoresis, while deflagellated cells did not.

  13. Drosophila roadblock and Chlamydomonas Lc7

    PubMed Central

    Bowman, Aaron B.; Patel-King, Ramila S.; Benashski, Sharon E.; McCaffery, J. Michael; Goldstein, Lawrence S.B.; King, Stephen M.

    1999-01-01

    Eukaryotic organisms utilize microtubule-dependent motors of the kinesin and dynein superfamilies to generate intracellular movement. To identify new genes involved in the regulation of axonal transport in Drosophila melanogaster, we undertook a screen based upon the sluggish larval phenotype of known motor mutants. One of the mutants identified in this screen, roadblock (robl), exhibits diverse defects in intracellular transport including axonal transport and mitosis. These defects include intra-axonal accumulations of cargoes, severe axonal degeneration, and aberrant chromosome segregation. The gene identified by robl encodes a 97–amino acid polypeptide that is 57% identical (70% similar) to the 105–amino acid Chlamydomonas outer arm dynein–associated protein LC7, also reported here. Both robl and LC7 have homology to several other genes from fruit fly, nematode, and mammals, but not Saccharomyces cerevisiae. Furthermore, we demonstrate that members of this family of proteins are associated with both flagellar outer arm dynein and Drosophila and rat brain cytoplasmic dynein. We propose that roadblock/LC7 family members may modulate specific dynein functions. PMID:10402468

  14. Radial spoke proteins of Chlamydomonas flagella

    PubMed Central

    Yang, Pinfen; Diener, Dennis R.; Yang, Chun; Kohno, Takahiro; Pazour, Gregory J.; Dienes, Jennifer M.; Agrin, Nathan S.; King, Stephen M.; Sale, Winfield S.; Kamiya, Ritsu; Rosenbaum, Joel L.; Witman, George B.

    2007-01-01

    Summary The radial spoke is a ubiquitous component of ‘9+2’ cilia and flagella, and plays an essential role in the control of dynein arm activity by relaying signals from the central pair of microtubules to the arms. The Chlamydomonas reinhardtii radial spoke contains at least 23 proteins, only 8 of which have been characterized at the molecular level. Here, we use mass spectrometry to identify 10 additional radial spoke proteins. Many of the newly identified proteins in the spoke stalk are predicted to contain domains associated with signal transduction, including Ca2+-, AKAP- and nucleotide-binding domains. This suggests that the spoke stalk is both a scaffold for signaling molecules and itself a transducer of signals. Moreover, in addition to the recently described HSP40 family member, a second spoke stalk protein is predicted to be a molecular chaperone, implying that there is a sophisticated mechanism for the assembly of this large complex. Among the 18 spoke proteins identified to date, at least 12 have apparent homologs in humans, indicating that the radial spoke has been conserved throughout evolution. The human genes encoding these proteins are candidates for causing primary ciliary dyskinesia, a severe inherited disease involving missing or defective axonemal structures, including the radial spokes. PMID:16507594

  15. Genetic tools and techniques for Chlamydomonas reinhardtii.

    PubMed

    Mussgnug, Jan H

    2015-07-01

    The development of tools has always been a major driving force for the advancement of science. Optical microscopes were the first instruments that allowed discovery and descriptive studies of the subcellular features of microorganisms. Although optical and electron microscopes remained at the forefront of microbiological research tools since their inventions, the advent of molecular genetics brought about questions which had to be addressed with new "genetic tools". The unicellular green microalgal genus Chlamydomonas, especially the most prominent species C. reinhardtii, has become a frequently used model organism for many diverse fields of research and molecular genetic analyses of C. reinhardtii, as well as the available genetic tools and techniques, have become increasingly sophisticated throughout the last decades. The aim of this review is to provide an overview of the molecular key features of C. reinhardtii and summarize the progress related to the development of tools and techniques for genetic engineering of this organism, from pioneering DNA transformation experiments to state-of-the-art techniques for targeted nuclear genome editing and high-throughput screening approaches.

  16. Characterization of cooperative bicarbonate uptake into chloroplast stroma in the green alga Chlamydomonas reinhardtii

    PubMed Central

    Yamano, Takashi; Sato, Emi; Iguchi, Hiro; Fukuda, Yuri; Fukuzawa, Hideya

    2015-01-01

    The supply of inorganic carbon (Ci; CO2 and HCO3–) is an environmental rate-limiting factor in aquatic photosynthetic organisms. To overcome the difficulty in acquiring Ci in limiting-CO2 conditions, an active Ci uptake system called the CO2-concentrating mechanism (CCM) is induced to increase CO2 concentrations in the chloroplast stroma. An ATP-binding cassette transporter, HLA3, and a formate/nitrite transporter homolog, LCIA, are reported to be associated with HCO3– uptake [Wang and Spalding (2014) Plant Physiol 166(4):2040–2050]. However, direct evidence of the route of HCO3– uptake from the outside of cells to the chloroplast stroma remains elusive owing to a lack of information on HLA3 localization and comparative analyses of the contribution of HLA3 and LCIA to the CCM. In this study, we revealed that HLA3 and LCIA are localized to the plasma membrane and chloroplast envelope, respectively. Insertion mutants of HLA3 and/or LCIA showed decreased Ci affinities/accumulation, especially in alkaline conditions where HCO3– is the predominant form of Ci. HLA3 and LCIA formed protein complexes independently, and the absence of LCIA decreased HLA3 mRNA accumulation, suggesting the presence of unidentified retrograde signals from the chloroplast to the nucleus to maintain HLA3 mRNA expression. Furthermore, although single overexpression of HLA3 or LCIA in high CO2 conditions did not affect Ci affinity, simultaneous overexpression of HLA3 with LCIA significantly increased Ci affinity/accumulation. These results highlight the HLA3/LCIA-driven cooperative uptake of HCO3– and a key role of LCIA in the maintenance of HLA3 stability as well as Ci affinity/accumulation in the CCM. PMID:26015566

  17. Direct transesterification of Oedogonium sp. oil be using immobilized isolated novel Bacillus sp. lipase.

    PubMed

    Sivaramakrishnan, Ramachandran; Muthukumar, Karuppan

    2014-01-01

    This work emphasizes the potential of the isolated Bacillus sp. lipase for the production of fatty acid methyl ester by the direct transesterification of Oedogonium sp. of macroalgae. Dimethyl carbonate was used as the extraction solvent and also as the reactant. The effect of solvent/algae ratio, water addition, catalyst, temperature, stirring and time on the direct transesterification was studied. The highest fatty acid methyl ester yield obtained under optimum conditions (5 g Oedogonium sp. powder, 7.5 ml of solvent (dimethyl carbonate)/g of algae, 8% catalyst (%wt/wt of oil), distilled water 1% (wt/wt of algae), 36 h, 55°C and 180 rpm) was 82%. Final product was subjected to thermogravimetric analysis and (1)H NMR analysis. The results showed that the isolated enzyme has good potential in catalyzing the direct transesterification of algae, and the dimethyl carbonate did not affect the activity of the isolated lipase.

  18. Swapped green algal promoters: aphVIII-based gene constructs with Chlamydomonas flanking sequences work as dominant selectable markers in Volvox and vice versa.

    PubMed

    Hallmann, A; Wodniok, S

    2006-06-01

    Production of transgenic organisms is a well-established, versatile course of action in molecular biology. Genetic engineering often requires heterologous, dominant antibiotic resistance genes that have been used as selectable markers in many species. However, as heterologous 5' and 3' flanking sequences often result in very low expression rates, endogenous flanking sequences, especially promoters, are mostly required and are easily obtained in model organisms, but it is much more complicated and time-consuming to get appropriate sequences from less common organisms. In this paper, we show that aminoglycoside 3'-phosphotransferase gene (aphVIII) based constructs with 3' and 5' untranslated flanking sequences (including promoters) from the multicellular green alga Volvox work in the unicellular green alga Chlamydomonas and flanking sequences from Chlamydomonas work in Volvox, at least if a low expression rate is compensated by an enforced high gene dosage. This strategy might be useful for all investigators that intend to transform species in which genomic sequences are not available, but sequences from related organisms exist.

  19. The occurrence of the psbS gene product in Chlamydomonas reinhardtii and in other photosynthetic organisms and its correlation with energy quenching.

    PubMed

    Bonente, Giulia; Passarini, Francesca; Cazzaniga, Stefano; Mancone, Carmine; Buia, Maria Cristina; Tripodi, Marco; Bassi, Roberto; Caffarri, Stefano

    2008-01-01

    To avoid photodamage, photosynthetic organisms have developed mechanisms to evade or dissipate excess energy. Lumen overacidification caused by light-induced electron transport triggers quenching of excited chlorophylls and dissipation of excess energy into heat. In higher plants participation of the PsbS protein as the sensor of low lumenal pH was clearly demonstrated. Although light-dependent energy quenching is a property of all photosynthetic organisms, large differences in amplitude and kinetics can be observed thus raising the question whether a single common mechanism is in action. We performed a detailed study of PsbS expression/accumulation in Chlamydomonas reinhardtii and investigated its accumulation in other algae and plants. We showed that PsbS cannot be detected in Chlamydomonas under a wide range of growth conditions. Overexpression of the endogenous psbs gene showed that the corresponding protein could not be addressed to the thylakoid membranes. Survey of different unicellular green algae showed no accumulation of anti-PsbS reactive proteins differently from multicellular species. Nevertheless, some unicellular species exhibit high energy quenching activity, suggesting that a PsbS-independent mechanism is activated. By correlating growth habitat and PsbS accumulation in different species, we suggest that during the evolution the light environment has been a determinant factor for the conservation/loss of the PsbS function.

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

    SciTech Connect

    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

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

    PubMed

    Ramos-Martinez, E M; Fimognari, L; Sakuragi, Y

    2017-02-16

    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. In order 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 folds, with the maximum yield of 15 mg L(-1) . Moreover, the presence of the glycomodules confererred 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. This article is protected by copyright. All rights reserved.

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

    NASA Astrophysics Data System (ADS)

    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

  3. Effects of nitrogen and phosphorus on arsenite accumulation, oxidation, and toxicity in Chlamydomonas reinhardtii.

    PubMed

    Wang, Ning-Xin; Huang, Bin; Xu, Shen; Wei, Zhong-Bo; Miao, Ai-Jun; Ji, Rong; Yang, Liu-Yan

    2014-12-01

    We studied arsenite (iAs(III)) accumulation, oxidation, and toxicity in the freshwater green alga Chlamydomonas reinhardtii under nutrient-enriched (+NP), phosphorus-limited (-P), and nitrogen-limited (-N) conditions. The -P alga (55.1 μM) had a Michaelis constant (Kd) for uptake approximately one tenth of the +NP (419 μM) and -N (501 μM) cells, indicating iAs(III) uptake inhibition by extracellular phosphate. This conclusion was supported by the hyperbolic reduction in iAs(III) uptake rate (V) from 9.2 to 0.8 μmol/g-dw/h when the extracellular phosphate concentration went up from 0 to 250 μM. The maximal iAs(III) uptake rate (Vmax) of the -N alga (24.3 μmol/g-dw/h) was twice as much as that of the +NP (12 μmol/g-dw/h) and -P (8.1 μmol/g-dw/h) cells. It implies that more arsenic transporters were synthesized under the -N condition. Once accumulated, iAs(III) was oxidized and a higher proportion of arsenate (iAs(V)) was observed at lower [As]dis or under nutrient-limited conditions. Nevertheless, iAs(III) oxidation mainly occurred outside the cells with the extent of oxidation reciprocal to [As]dis. Based on the logistic modeling of the concentration-response curves in the +NP, -P, and -N toxicity tests, iAs(III) had an [As]dis-based EC50 of 1763, 13.1, and 1208 μM and an intracellular arsenic concentration based EC50 of 35.6, 28.8, and 195 μmol/g-dw, respectively. Higher iAs(III) toxicity to the -P cells occured because of their increased iAs(III) accumulation, whereas the underlying mechanisms why the -N alga was more tolerant need to be further revealed. Overall, both N and P had remarkable effects on the behavior and effects of iAs(III), which cannot be disregarded in the biogeochemical cycling research of arsenic.

  4. Genomic analysis of organismal complexity in the multicellular green alga Volvox carteri.

    PubMed

    Prochnik, Simon E; Umen, James; Nedelcu, Aurora M; 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, Rüdiger; Kirk, David; Rokhsar, Daniel S

    2010-07-09

    The multicellular green alga Volvox carteri and its morphologically diverse close relatives (the volvocine algae) are well suited for the investigation of the evolution of multicellularity and development. We sequenced the 138-mega-base pair genome of V. carteri and compared its approximately 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. Volvox is enriched in volvocine-algal-specific proteins, 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.

  5. Converting carbohydrates extracted from marine algae into ethanol using various ethanolic Escherichia coli strains.

    PubMed

    Lee, Soojin; Oh, Younghoon; Kim, Donghyun; Kwon, Doyeon; Lee, Choulgyun; Lee, Jinwon

    2011-07-01

    Marine algae, which make up about 80% of the world's living organisms, contain many energy sources, such as sugars and lipids. Therefore, the possibility of utilizing structural carbohydrates from marine algae for bioethanol production has been studied. In order to obtain monosaccharides, Undaria pinnatifida, Chlorella vulgaris, and Chlamydomonas reinhardtii were used for the saccharification experiments. The pretreatment was carried out by dilute acid hydrolysis and enzymatic treatment. To find the optimal conditions, experiments were performed at several temperatures, acid concentrations, pH conditions and durations. To test bioethanol production, several ethanolic E. coli W3110 strains, which were developed previously, were used. The maximum yield of bioethanol, 0.4 g ethanol/g biomass, was achieved with pretreated C. vulgaris and E. coli SJL2526, derived from wild-type E. coli W3110 and which includes the adhB, pdc, galP, and glk genes.

  6. L,L-Diaminopimelate Aminotransferase from Chlamydomonas reinhardtii: A Target for Algaecide Development

    PubMed Central

    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. PMID:21633707

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

    PubMed

    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.

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

    PubMed

    Hemschemeier, Anja; Fouchard, Swanny; Cournac, Laurent; Peltier, Gilles; Happe, Thomas

    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 H2 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 H2 producers. Two of the still most disputed questions regarding H2 generation by C. reinhardtii concern the electron source for H2 evolution and the competition of the hydrogenase with alternative electron sinks. We analyzed the H2 metabolism of S-depleted C. reinhardtii cultures utilizing a special mass spectrometer setup and investigated the influence of photosystem II (PSII)- or ribulosebisphosphate-carboxylase/oxygenase (Rubisco)-deficiency. We show that electrons for H2-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 H2 also in the presence of sulfur, H2 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 H2 evolution.

  9. Experimental Definition and Validation of Protein Coding Transcripts in Chlamydomonas reinhardtii

    SciTech Connect

    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

  10. The Fox1 Ferroxidase of Chlamydomonas reinhardtii: A New Multicopper Oxidase Structural Paradigm

    PubMed Central

    Terzulli, Alaina J.; Kosman, Daniel J.

    2009-01-01

    Multicopper oxidases (MCO) contain at least four copper atoms arrayed in three distinct ligand fields supported by two canonical structural features: 1) multiples of the cupredoxin fold; and 2) four unique sequence elements that include the 10 histidine and one cysteine ligands to the four copper atoms. Ferroxidases are a sub-family of MCO proteins that contain residues supporting a specific reactivity towards ferrous iron; these MCOs play a vital role in iron metabolism in bacteria, algae, fungi and mammals. In contrast to the fungal ferroxidases, e.g. Fet3p from Saccharomyces cerevisiae, the mammalian ceruloplasmin (Cp) is twice as large (six versus three cupredoxin domains) and contains three type 1, or “blue” copper sites. Chlamydomonas reinhardtii expresses a putative ferroxidase, Fox1, which has sequence similarity to human Cp (hCp). Eschewing the standard sequence-based modeling paradigm, a function-based model of the Fox1 protein has been constructed which replicates hCp's six Cu-site ligand arrays with an overall root mean square deviation of 1.4 Å. Analysis of this model has led also to assignment of motifs in Fox1 that are unique to ferroxidases, the strongest evidence to date that the well-characterized fungal high-affinity iron uptake system is essential to iron homeostasis in green algae. The model of Fox1 also establishes a sub-family of MCO proteins with a non-canonical Cu-ligand organization. These diverse structures suggest alternate mechanisms for intramolecular electron transfer and require a new trajectory for the evolution of the MCO superfamily. PMID:19023602

  11. MicroRNAs modulate adaption to multiple abiotic stresses in Chlamydomonas reinhardtii

    PubMed Central

    Gao, Xiang; Zhang, Fengge; Hu, Jinlu; Cai, Wenkai; Shan, Ge; Dai, Dongsheng; Huang, Kaiyao; Wang, Gaohong

    2016-01-01

    MicroRNAs play an important role in abiotic stress responses in higher plants and animals, but their role in stress adaptation in algae remains unknown. In this study, the expression of identified and putative miRNAs in Chlamydomonas reinhardtii was assessed using quantitative polymerase chain reaction; some of the miRNAs (Cre-miR906-3p) were up-regulated, whereas others (Cre-miR910) were down-regulated when the species was subjected to multiple abiotic stresses. With degradome sequencing data, we also identified ATP4 (the d-subunit of ATP synthase) and NCR2 (NADPH: cytochrome P450 reductase) as one of the several targets of Cre-miR906-3p and Cre-miR910, respectively. Q-PCR data indicated that ATP4, which was expressed inversely in relation to Cre-miR906-3p under stress conditions. Overexpressing of Cre-miR906-3p enhanced resistance to multiple stresses; conversely, overexpressing of ATP4 produced the opposite effect. These data of Q-PCR, degradome sequencing and adaptation of overexpressing lines indicated that Cre-miR906-3p and its target ATP4 were a part of the same pathway for stress adaptation. We found that Cre-miR910 and its target NCR2 were also a part of this pathway. Overexpressing of Cre-miR910 decreased, whereas that of NCR2 increased the adaption to multiple stresses. Our findings suggest that the two classes of miRNAs synergistically mediate stress adaptation in algae. PMID:27910907

  12. Visualization of oxygen distribution patterns caused by coral and algae.

    PubMed

    Haas, Andreas F; Gregg, Allison K; Smith, Jennifer E; Abieri, Maria L; Hatay, Mark; Rohwer, Forest

    2013-01-01

    Planar optodes were used to visualize oxygen distribution patterns associated with a coral reef associated green algae (Chaetomorpha sp.) and a hermatypic coral (Favia sp.) separately, as standalone organisms, and placed in close proximity mimicking coral-algal interactions. Oxygen patterns were assessed in light and dark conditions and under varying flow regimes. The images show discrete high oxygen concentration regions above the organisms during lighted periods and low oxygen in the dark. Size and orientation of these areas were dependent on flow regime. For corals and algae in close proximity the 2D optodes show areas of extremely low oxygen concentration at the interaction interfaces under both dark (18.4 ± 7.7 µmol O2 L(- 1)) and daylight (97.9 ± 27.5 µmol O2 L(- 1)) conditions. These images present the first two-dimensional visualization of oxygen gradients generated by benthic reef algae and corals under varying flow conditions and provide a 2D depiction of previously observed hypoxic zones at coral algae interfaces. This approach allows for visualization of locally confined, distinctive alterations of oxygen concentrations facilitated by benthic organisms and provides compelling evidence for hypoxic conditions at coral-algae interaction zones.

  13. Independent Control of the Static and Dynamic Components of the Chlamydomonas Flagellar Beat.

    PubMed

    Geyer, Veikko F; Sartori, Pablo; Friedrich, Benjamin M; Jülicher, Frank; Howard, Jonathon

    2016-04-25

    When the green alga Chlamydomonas reinhardtii swims, it uses the breaststroke beat of its two flagella to pull itself forward [1]. The flagellar waveform can be decomposed into a static component, corresponding to an asymmetric time-averaged shape, and a dynamic component, corresponding to the time-varying wave [2]. Extreme lightening conditions photoshock the cell, converting the breaststroke beat into a symmetric sperm-like beat, which causes a reversal of the direction of swimming [3]. Waveform conversion is achieved by a reduction in magnitude of the static component, whereas the dynamic component remains unchanged [2]. The coupling between static and dynamic components, however, is poorly understood, and it is not known whether the static component requires the dynamic component or whether it can exist independently. We used isolated and reactivated axonemes [4] to investigate the relation between the two beat components. We discovered that, when reactivated in the presence of low ATP concentrations, axonemes displayed the static beat component in absence of the dynamic component. Furthermore, we found that the amplitudes of the two components depend on ATP in qualitatively different ways. These results show that the decomposition into static and dynamic components is not just a mathematical concept but that the two components can independently control different aspects of cell motility: the static component controls swimming direction, whereas the dynamic component provides propulsion.

  14. An organelle K+ channel is required for osmoregulation in Chlamydomonas reinhardtii.

    PubMed

    Xu, Feifei; Wu, Xiaoan; Jiang, Lin-Hua; Zhao, Hucheng; Pan, Junmin

    2016-08-01

    Fresh water protozoa and algae face hypotonic challenges in their living environment. Many of them employ a contractile vacuole system to uptake excessive water from the cytoplasm and expel it to the environment to achieve cellular homeostasis. K(+), a major osmolyte in contractile vacuole, is predicted to create higher osmolarity for water influx. Molecular mechanisms for K(+) permeation through the plasma membrane have been well studied. However, how K(+) permeates organelles such as the contractile vacuole is not clear. Here, we show that the six-transmembrane K(+) channel KCN11 in Chlamydomonas is exclusively localized to contractile vacuole. Ectopic expression of KCN11 in HEK293T cells results in voltage-gated K(+) channel activity. Disruption of the gene or mutation of key residues for K(+) permeability of the channel leads to dysfunction of cell osmoregulation in very hypotonic conditions. The contractile cycle is inhibited in the mutant cells with a slower rate of contractile vacuole swelling, leading to cell death. These data demonstrate a new role for six-transmembrane K(+) channels in contractile vacuole functioning and provide further insights into osmoregulation mediated by the contractile vacuole.

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

    PubMed

    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.

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

    PubMed Central

    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

  17. Temperature-Sensitive, Photosynthesis-Deficient Mutants of Chlamydomonas reinhardtii1

    PubMed Central

    Spreitzer, Robert J.; Al-Abed, Souhail R.; Huether, Michael J.

    1988-01-01

    Mutants of the unicellular, green alga Chlamydomonas reinhardtii were recovered by screening for the absence of photoautotrophic growth at 35°C. Whereas nonconditional mutants required acetate for growth at both 25 and 35°C, the conditional mutants have normal photoautotrophic growth at 25°C. The conditional mutants consisted of two classes: (a) Temperature-sensitive mutants died under all growth conditions at 35°C, but (b) temperature-sensitive, acetate-requiring mutants were capable of heterotrophic growth at 35°C when supplied with acetate in the dark. The majority of mutants within the latter of these two classes had defects in photosynthetic functions. These defects included altered pigmentation, reduced whole-chain electron-transport activity, reduced ribulosebis-phosphate carboxylase activity, or pleiotropic alterations in a number of these photosynthetic components. Both nuclear and chloroplast mutants were identified, and a correlation between light-sensitive and photosynthesis-deficient phenotypes was observed. PMID:16665986

  18. Proteomic Analysis of a Fraction with Intact Eyespots of Chlamydomonas reinhardtii and Assignment of Protein Methylation

    PubMed Central

    Eitzinger, Nicole; Wagner, Volker; Weisheit, Wolfram; Geimer, Stefan; Boness, David; Kreimer, Georg; Mittag, Maria

    2015-01-01

    Flagellate green algae possess a visual system, the eyespot. In Chlamydomonas reinhardtii it is situated at the edge of the chloroplast and consists of two carotenoid rich lipid globule layers subtended by thylakoid membranes (TM) that are attached to both chloroplast envelope membranes and a specialized area of the plasma membrane (PM). A former analysis of an eyespot fraction identified 203 proteins. To increase the understanding of eyespot related processes, knowledge of the protein composition of the membranes in its close vicinity is desirable. Here, we present a purification procedure that allows isolation of intact eyespots. This gain in intactness goes, however, hand in hand with an increase of contaminants from other organelles. Proteomic analysis identified 742 proteins. Novel candidates include proteins for eyespot development, retina-related proteins, ion pumps, and membrane-associated proteins, calcium sensing proteins as well as kinases, phosphatases and 14-3-3 proteins. Methylation of proteins at Arg or Lys is known as an important posttranslational modification involved in, e.g., signal transduction. Here, we identify several proteins from eyespot fractions that are methylated at Arg and/or Lys. Among them is the eyespot specific SOUL3 protein that influences the size and position of the eyespot and EYE2, a protein important for its development. PMID:26697039

  19. A small multifunctional pentatricopeptide repeat protein in the chloroplast of Chlamydomonas reinhardtii.

    PubMed

    Jalal, Abdullah; Schwarz, Christian; Schmitz-Linneweber, Christian; Vallon, Olivier; Nickelsen, Jörg; Bohne, Alexandra-Viola

    2015-03-01

    Organellar biogenesis is mainly regulated by nucleus-encoded factors, which act on various steps of gene expression including RNA editing, processing, splicing, stabilization, and translation initiation. Among these regulatory factors, pentatricopeptide repeat (PPR) proteins form the largest family of RNA binding proteins, with hundreds of members in flowering plants. In striking contrast, the genome of the unicellular green alga Chlamydomonas reinhardtii encodes only 14 such proteins. In this study, we analyzed PPR7, the smallest and most highly expressed PPR protein in C. reinhardtii. Green fluorescent protein-based localization and gel-filtration analysis revealed that PPR7 forms a part of a high-molecular-weight ribonucleoprotein complex in the chloroplast stroma. RIP-chip analysis of PPR7-bound RNAs demonstrated that the protein associates with a diverse set of chloroplast transcripts in vivo, i.e. rrnS, psbH, rpoC2, rbcL, atpA, cemA-atpH, tscA, and atpI-psaJ. Furthermore, the investigation of PPR7 RNAi strains revealed that depletion of PPR7 results in a light-sensitive phenotype, accompanied by altered levels of its target RNAs that are compatible with the defects in their maturation or stabilization. PPR7 is thus an unusual type of small multifunctional PPR protein, which interacts, probably in conjunction with other RNA binding proteins, with numerous target RNAs to promote a variety of post-transcriptional events.

  20. Phosphoprotein SAK1 is a regulator of acclimation to singlet oxygen in Chlamydomonas reinhardtii

    PubMed Central

    Wakao, Setsuko; Chin, Brian L; Ledford, Heidi K; Dent, Rachel M; Casero, David; Pellegrini, Matteo; Merchant, Sabeeha S; Niyogi, Krishna K

    2014-01-01

    Singlet oxygen is a highly toxic and inevitable byproduct of oxygenic photosynthesis. The unicellular green alga Chlamydomonas reinhardtii is capable of acclimating specifically to singlet oxygen stress, but the retrograde signaling pathway from the chloroplast to the nucleus mediating this response is unknown. Here we describe a mutant, singlet oxygen acclimation knocked-out 1 (sak1), that lacks the acclimation response to singlet oxygen. Analysis of genome-wide changes in RNA abundance during acclimation to singlet oxygen revealed that SAK1 is a key regulator of the gene expression response during acclimation. The SAK1 gene encodes an uncharacterized protein with a domain conserved among chlorophytes and present in some bZIP transcription factors. The SAK1 protein is located in the cytosol, and it is induced and phosphorylated upon exposure to singlet oxygen, suggesting that it is a critical intermediate component of the retrograde signal transduction pathway leading to singlet oxygen acclimation. DOI: http://dx.doi.org/10.7554/eLife.02286.001 PMID:24859755

  1. Phosphoprotein SAK1 is a regulator of acclimation to singlet oxygen in Chlamydomonas reinhardtii.

    PubMed

    Wakao, Setsuko; Chin, Brian L; Ledford, Heidi K; Dent, Rachel M; Casero, David; Pellegrini, Matteo; Merchant, Sabeeha S; Niyogi, Krishna K

    2014-05-23

    Singlet oxygen is a highly toxic and inevitable byproduct of oxygenic photosynthesis. The unicellular green alga Chlamydomonas reinhardtii is capable of acclimating specifically to singlet oxygen stress, but the retrograde signaling pathway from the chloroplast to the nucleus mediating this response is unknown. Here we describe a mutant, singlet oxygen acclimation knocked-out 1 (sak1), that lacks the acclimation response to singlet oxygen. Analysis of genome-wide changes in RNA abundance during acclimation to singlet oxygen revealed that SAK1 is a key regulator of the gene expression response during acclimation. The SAK1 gene encodes an uncharacterized protein with a domain conserved among chlorophytes and present in some bZIP transcription factors. The SAK1 protein is located in the cytosol, and it is induced and phosphorylated upon exposure to singlet oxygen, suggesting that it is a critical intermediate component of the retrograde signal transduction pathway leading to singlet oxygen acclimation.DOI: http://dx.doi.org/10.7554/eLife.02286.001.

  2. Asymmetric properties of the Chlamydomonas reinhardtii cytoskeleton direct rhodopsin photoreceptor localization

    PubMed Central

    Mittelmeier, Telsa M.; Boyd, Joseph S.; Lamb, Mary Rose

    2011-01-01

    The eyespot of the unicellular green alga Chlamydomonas reinhardtii is a photoreceptive organelle required for phototaxis. Relative to the anterior flagella, the eyespot is asymmetrically positioned adjacent to the daughter four-membered rootlet (D4), a unique bundle of acetylated microtubules extending from the daughter basal body toward the posterior of the cell. Here, we detail the relationship between the rhodopsin eyespot photoreceptor Channelrhodopsin 1 (ChR1) and acetylated microtubules. In wild-type cells, ChR1 was observed in an equatorial patch adjacent to D4 near the end of the acetylated microtubules and along the D4 rootlet. In cells with cytoskeletal protein mutations, supernumerary ChR1 patches remained adjacent to acetylated microtubules. In mlt1 (multieyed) mutant cells, supernumerary photoreceptor patches were not restricted to the D4 rootlet, and more anterior eyespots correlated with shorter acetylated microtubule rootlets. The data suggest a model in which photoreceptor localization is dependent on microtubule-based trafficking selective for the D4 rootlet, which is perturbed in mlt1 mutant cells. PMID:21555459

  3. The Awesome Power of Dikaryons for Studying Flagella and Basal Bodies in Chlamydomonas reinhardtii

    PubMed Central

    Dutcher, Susan K.

    2014-01-01

    Cilia/flagella and basal bodies/centrioles play key roles in human health and homeostasis. Among the organisms used to study these microtubule-based organelles, the green alga Chlamydomonas reinhardtii has several advantages. One is the existence of a temporary phase of the life cycle, termed the dikaryon. These cells are formed during mating when the cells fuse and the behavior of flagella from two genetically distinguishable parents can be observed. During this stage, the cytoplasms mix allowing for a defect in the flagella of one parent to be rescued by proteins from the other parent. This offers the unique advantage of adding back wild-type gene product or labeled protein at endogenous levels that can used to monitor various flagellar and basal body phenotypes. Mutants that show rescue and ones that fail to show rescue are both informative about the nature of the flagella and basal body defects. When rescue occurs, it can be used to determine the mutant gene product and to follow the temporal and spatial patterns of flagellar assembly. This review describes many examples of insights into basal body and flagellar proteins’ function and assembly that have been discovered using dikaryons and discusses the potential for further analyses. PMID:24272949

  4. Dichromate effect on energy dissipation of photosystem II and photosystem I in Chlamydomonas reinhardtii.

    PubMed

    Perreault, François; Ait Ali, Nadia; Saison, Cyril; Popovic, Radovan; Juneau, Philippe

    2009-07-17

    In this study, we investigated the energy dissipation processes via photosystem II and photosystem I activity in green alga Chlamydomonas reinhardtii exposed to dichromate inhibitory effect. Quantum yield of photosystem II and also photosystem I were highly decreased by dichromate effect. Such inhibition by dichromate induced strong quenching effect on rapid OJIP fluorescence transients, indicating deterioration of photosystem II electron transport via plastoquinone pool toward photosystem I. The decrease of energy dissipation dependent on electron transport of photosystem II and photosystem I by dichromate effect was associated with strong increase of non-photochemical energy dissipation processes. By showing strong effect of dichromate on acceptor side of photosystem I, we indicated that dichromate inhibitory effect was not associated only with PSII electron transport. Here, we found that energy dissipation via photosystem I was limited by its electron acceptor side. By the analysis of P700 oxido-reduction state with methylviolagen as an exogenous PSI electron transport mediator, we showed that PSI electron transport discrepancy induced by dichromate effect was also caused by inhibitory effect located beyond photosystem I. Therefore, these results demonstrated that dichromate has different sites of inhibition which are associated with photosystem II, photosystem I and electron transport sink beyond photosystems.

  5. Adaptation of Chlamydomonas reinhardtii high-CO sub 2 -requiring mutants to limiting CO sub 2

    SciTech Connect

    Suzuki, K.; Spalding, M.H. )

    1989-07-01

    Photosynthetic characteristics of four high-CO{sub 2}-requiring mutants of Chlamydomonas reinhardtii were compared to those of wild type before and after a 24-hour exposure to limiting CO{sub 2} concentrations. The four mutants represent two loci involved in the CO{sub 2}-concentrating system of this unicellular alga. All mutants had a lower photosynthetic affinity for inorganic carbon than did the wild type when grown at an elevated CO{sub 2} concentration, indicating that the genetic lesion in each is expressed even at elevated CO{sub 2} concentrations. Wild type and all four mutants exhibited adaptive responses to limiting CO{sub 2} characteristic of the induction of the CO{sub 2}-concentrating system, resulting in an increased affinity for inorganic carbon only in wild type. Although other components of the CO{sub 2}-concentrating system were induced in these mutants, the defective component in each was sufficient to prevent any increase in the affinity for inorganic carbon. It was concluded that the genes corresponding to the ca-1 and pmp-1 loci exhibit at least partially constitutive expression and that all components of the CO{sub 2}-concentrating system may be required to significantly affect the photosynthetic affinity for inorganic carbon.

  6. Light Intensity is Important for Hydrogen Production in NaHSO3-Treated Chlamydomonas reinhardtii.

    PubMed

    Wei, Lanzhen; Yi, Jing; Wang, Lianjun; Huang, Tingting; Gao, Fudan; Wang, Quanxi; Ma, Weimin

    2017-01-07

    Chlamydomonas reinhardtii is a unicellular green alga that can use light energy to produce H2 from H2O in the background of NaHSO3 treatment. However, the role of light intensity in such H2 production remains elusive. Here, light intensity significantly affected the yield of H2 production in NaHSO3-treated C. reinhardtii, which was consistent with its effects on the content of O2 and the expression and activity of hydrogenase. Further, NaHSO3 was found to be able to remove O2 via a reaction of bisulfite with superoxide anion produced at the acceptor side of PSI, and light intensity affected the reaction rate significantly. Accordingly, high light and strong light but not low light can create an anaerobic environment, which is important to activate hydrogenase and produce H2 Based on the above results, we conclude that light intensity plays an important role in removing O2 and consequently activating hydrogenase and producing H2 in NaHSO3-treated C. reinhardtii.

  7. Evidence for phenotypic plasticity in the Antarctic extremophile Chlamydomonas raudensis Ettl. UWO 241.

    PubMed

    Pocock, Tessa; Vetterli, Adrien; Falk, Stefan

    2011-01-01

    Life in extreme environments poses unique challenges to photosynthetic organisms. The ability for an extremophilic green alga and its genetic and mesophilic equivalent to acclimate to changes in their environment was examined to determine the extent of their phenotypic plasticities. The Antarctic extremophile Chlamydomonas raudensis Ettl. UWO 241 (UWO) was isolated from an ice-covered lake in Antarctica, whereas its mesophilic counterpart C. raudensis Ettl. SAG 49.72 (SAG) was isolated from a meadow pool in the Czech Republic. The effects of changes in temperature and salinity on growth, morphology, and photochemistry were examined in the two strains. Differential acclimative responses were observed in UWO which include a wider salinity range for growth, and broader temperature- and salt-induced fluctuations in F(v)/F(m), relative to SAG. Furthermore, the redox state of the photosynthetic electron transport chain, measured as 1-q(P), was modulated in the extremophile whereas this was not observed in the mesophile. Interestingly, it is shown for the first time that SAG is similar to UWO in that it is unable to undergo state transitions. The different natural histories of these two strains exert different evolutionary pressures and, consequently, different abilities for acclimation, an important component of phenotypic plasticity. In contrast to SAG, UWO relied on a redox sensing and signalling system under the growth conditions used in this study. It is proposed that growth and adaptation of UWO under a stressful and extreme environment poises this extremophile for better success under changing environmental conditions.

  8. Application of Phosphoproteomics to Find Targets of Casein Kinase 1 in the Flagellum of Chlamydomonas

    PubMed Central

    Boesger, Jens; Wagner, Volker; Weisheit, Wolfram; Mittag, Maria

    2012-01-01

    The green biflagellate alga Chlamydomonas reinhardtii serves as model for studying structural and functional features of flagella. The axoneme of C. reinhardtii anchors a network of kinases and phosphatases that control motility. One of them, Casein Kinase 1 (CK1), is known to phosphorylate the Inner Dynein Arm I1 Intermediate Chain 138 (IC138), thereby regulating motility. CK1 is also involved in regulating the circadian rhythm of phototaxis and is relevant for the formation of flagella. By a comparative phosphoproteome approach, we determined phosphoproteins in the flagellum that are targets of CK1. Thereby, we applied the specific CK1 inhibitor CKI-7 that causes significant changes in the flagellum phosphoproteome and reduces the swimming velocity of the cells. In the CKI-7-treated cells, 14 phosphoproteins were missing compared to the phosphoproteome of untreated cells, including IC138, and four additional phosphoproteins had a reduced number of phosphorylation sites. Notably, inhibition of CK1 causes also novel phosphorylation events, indicating that it is part of a kinase network. Among them, Glycogen Synthase Kinase 3 is of special interest, because it is involved in the phosphorylation of key clock components in flies and mammals and in parallel plays an important role in the regulation of assembly in the flagellum. PMID:23316220

  9. Application of phosphoproteomics to find targets of casein kinase 1 in the flagellum of chlamydomonas.

    PubMed

    Boesger, Jens; Wagner, Volker; Weisheit, Wolfram; Mittag, Maria

    2012-01-01

    The green biflagellate alga Chlamydomonas reinhardtii serves as model for studying structural and functional features of flagella. The axoneme of C. reinhardtii anchors a network of kinases and phosphatases that control motility. One of them, Casein Kinase 1 (CK1), is known to phosphorylate the Inner Dynein Arm I1 Intermediate Chain 138 (IC138), thereby regulating motility. CK1 is also involved in regulating the circadian rhythm of phototaxis and is relevant for the formation of flagella. By a comparative phosphoproteome approach, we determined phosphoproteins in the flagellum that are targets of CK1. Thereby, we applied the specific CK1 inhibitor CKI-7 that causes significant changes in the flagellum phosphoproteome and reduces the swimming velocity of the cells. In the CKI-7-treated cells, 14 phosphoproteins were missing compared to the phosphoproteome of untreated cells, including IC138, and four additional phosphoproteins had a reduced number of phosphorylation sites. Notably, inhibition of CK1 causes also novel phosphorylation events, indicating that it is part of a kinase network. Among them, Glycogen Synthase Kinase 3 is of special interest, because it is involved in the phosphorylation of key clock components in flies and mammals and in parallel plays an important role in the regulation of assembly in the flagellum.

  10. RNA interference silencing of a major lipid droplet protein affects lipid droplet size in Chlamydomonas reinhardtii.

    PubMed

    Moellering, Eric R; Benning, Christoph

    2010-01-01

    Eukaryotic cells store oils in the chemical form of triacylglycerols in distinct organelles, often called lipid droplets. These dynamic storage compartments have been intensely studied in the context of human health and also in plants as a source of vegetable oils for human consumption and for chemical or biofuel feedstocks. Many microalgae accumulate oils, particularly under conditions limiting to growth, and thus have gained renewed attention as a potentially sustainable feedstock for biofuel production. However, little is currently known at the cellular or molecular levels with regard to oil accumulation in microalgae, and the structural proteins and enzymes involved in the biogenesis, maintenance, and degradation of algal oil storage compartments are not well studied. Focusing on the model green alga Chlamydomonas reinhardtii, the accumulation of triacylglycerols and the formation of lipid droplets during nitrogen deprivation were investigated. Mass spectrometry identified 259 proteins in a lipid droplet-enriched fraction, among them a major protein, tentatively designated major lipid droplet protein (MLDP). This protein is specific to the green algal lineage of photosynthetic organisms. Repression of MLDP gene expression using an RNA interference approach led to increased lipid droplet size, but no change in triacylglycerol content or metabolism was observed.

  11. High-throughput fluorescence-activated cell sorting for lipid hyperaccumulating Chlamydomonas reinhardtii mutants.

    PubMed

    Xie, Bo; Stessman, Dan; Hart, Jason H; Dong, Haili; Wang, Yingjun; Wright, David A; Nikolau, Basil J; Spalding, Martin H; Halverson, Larry J

    2014-09-01

    The genetically tractable microalga Chlamydomonas reinhardtii has many advantages as a model for renewable bioproducts and/or biofuels production. However, one limitation of C. reinhardtii is its relatively low-lipid content compared with some other algal species. To overcome this limitation, we combined ethane methyl sulfonate mutagenesis with fluorescence-activated cell sorting (FACS) of cells stained with the lipophilic stain Nile Red to isolate lipid hyperaccumulating mutants of C. reinhardtii. By manipulating the FACS gates, we sorted mutagenized cells with extremely high Nile Red fluorescence signals that were rarely detected in nonmutagenized populations. This strategy successfully isolated several putative lipid hyperaccumulating mutants exhibiting 23% to 58% (dry weight basis) higher fatty acid contents than their progenitor strains. Significantly, for most mutants, nitrogen starvation was not required to attain high-lipid content nor was there a requirement for a deficiency in starch accumulation. Microscopy of Nile Red stained cells revealed that some mutants exhibit an increase in the number of lipid bodies, which correlated with TLC analysis of triacyglycerol content. Increased lipid content could also arise through increased biomass production. Collectively, our findings highlight the ability to enhance intracellular lipid accumulation in algae using random mutagenesis in conjunction with a robust FACS and lipid yield verification regime. Our lipid hyperaccumulating mutants could serve as a genetic resource for stacking additional desirable traits to further increase lipid production and for identifying genes contributing to lipid hyperaccumulation, without lengthy lipid-induction periods.

  12. Eyespot-dependent determination of the phototactic sign in Chlamydomonas reinhardtii

    PubMed Central

    Ueki, Noriko; Ide, Takahiro; Mochiji, Shota; Kobayashi, Yuki; Tokutsu, Ryutaro; Ohnishi, Norikazu; Yamaguchi, Katsushi; Shigenobu, Shuji; Tanaka, Kan; Minagawa, Jun; Hisabori, Toru; Hirono, Masafumi; Wakabayashi, Ken-ichi

    2016-01-01

    The biflagellate green alga Chlamydomonas reinhardtii exhibits both positive and negative phototaxis to inhabit areas with proper light conditions. It has been shown that treatment of cells with reactive oxygen species (ROS) reagents biases the phototactic sign to positive, whereas that with ROS scavengers biases it to negative. Taking advantage of this property, we isolated a mutant, lts1-211, which displays a reduction-oxidation (redox) dependent phototactic sign opposite to that of the wild type. This mutant has a single amino acid substitution in phytoene synthase, an enzyme that functions in the carotenoid-biosynthesis pathway. The eyespot contains large amounts of carotenoids and is crucial for phototaxis. Most lts1-211 cells have no detectable eyespot and reduced carotenoid levels. Interestingly, the reversed phototactic-sign phenotype of lts1-211 is shared by other eyespot-less mutants. In addition, we directly showed that the cell body acts as a convex lens. The lens effect of the cell body condenses the light coming from the rear onto the photoreceptor in the absence of carotenoid layers, which can account for the reversed-phototactic-sign phenotype of the mutants. These results suggest that light-shielding property of the eyespot is essential for determination of phototactic sign. PMID:27122315

  13. Genetic analysis of suppressors of the PF10 mutation in Chlamydomonas reinhardtii

    SciTech Connect

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

    1988-12-01

    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.

  14. The awesome power of dikaryons for studying flagella and basal bodies in Chlamydomonas reinhardtii.

    PubMed

    Dutcher, Susan K

    2014-02-01

    Cilia/flagella and basal bodies/centrioles play key roles in human health and homeostasis. Among the organisms used to study these microtubule-based organelles, the green alga Chlamydomonas reinhardtii has several advantages. One is the existence of a temporary phase of the life cycle, termed the dikaryon. These cells are formed during mating when the cells fuse and the behavior of flagella from two genetically distinguishable parents can be observed. During this stage, the cytoplasms mix allowing for a defect in the flagella of one parent to be rescued by proteins from the other parent. This offers the unique advantage of adding back wild-type gene product or labeled protein at endogenous levels that can used to monitor various flagellar and basal body phenotypes. Mutants that show rescue and ones that fail to show rescue are both informative about the nature of the flagella and basal body defects. When rescue occurs, it can be used to determine the mutant gene product and to follow the temporal and spatial patterns of flagellar assembly. This review describes many examples of insights into basal body and flagellar proteins' function and assembly that have been discovered using dikaryons and discusses the potential for further analyses.

  15. Acetic acid-induced programmed cell death and release of volatile organic compounds in Chlamydomonas reinhardtii.

    PubMed

    Zuo, Zhaojiang; Zhu, Yerong; Bai, Yanling; Wang, Yong

    2012-02-01

    Acetic acid widely spreads in atmosphere, aquatic ecosystems containing residues and anoxic soil. It can inhibit aquatic plant germination and growth, and even cause programmed cell death (PCD) of yeast. In the present study, biochemical and physiological responses of the model unicellular green algae Chlamydomonas reinhardtii were examined after acetic acid stress. H(2)O(2) burst was found in C. reinhardtii after acetic acid stress at pH 5.0 for 10 min. The photosynthetic pigments were degraded, gross photosynthesis and respiration were disappeared gradually, and DNA fragmentation was also detected. Those results indicated that C. reinhardtii cells underwent a PCD but not a necrotic, accidental cell death event. It was noticed that C. reinhardtii cells in PCD released abundant volatile organic compounds (VOCs) upon acetic acid stress. Therefore, we analyzed the VOCs and tested their effects on other normal cells. The treatment of C. reinhardtii cultures with VOCs reduced the cell density and increased antioxidant enzyme activity. Therefore, a function of VOCs as infochemicals involved in cell-to-cell communication at the conditions of applied stress is suggested.

  16. Two equilibration pools of chlorophylls in the Photosystem I core antenna of Chlamydomonas reinhardtii.

    PubMed

    Gibasiewicz, Krzysztof; Ramesh, V M; Lin, Su; Redding, Kevin; Woodbury, Neal W; Webber, Andrew N

    2007-04-01

    Femtosecond transient absorption spectroscopy was applied for a comparative study of excitation decay in several different Photosystem I (PSI) core preparations from the green alga Chlamydomonas reinhardtii. For PSI cores with a fully interconnected network of chlorophylls, the excitation energy was equilibrated over a pool of chlorophylls absorbing at approximately 683 nm, independent of excitation wavelength [Gibasiewicz et al. J Phys Chem B 105:11498-11506, 2001; J Phys Chem B 106:6322-6330, 2002]. In preparations with impaired connectivity between chlorophylls, we have found that the spectrum of chlorophylls connected to the reaction center (i.e., with approximately 20 ps decay time) over which the excitation is equilibrated becomes excitation-wavelength-dependent. Excitation at 670 nm is finally equilibrated over chlorophylls absorbing at approximately 675 nm, whereas excitation at 695 nm or 700 nm is equilibrated over chlorophylls absorbing at approximately 683 nm. This indicates that in the vicinity of the reaction center there are two spectrally different and spatially separated pools of chlorophylls that are equally capable of effective excitation energy transfer to the reaction center. We propose that they are related to the two groups of central PSI core chlorophylls lying on the opposite sides of reaction center.

  17. The Involvement of hybrid cluster protein 4, HCP4, in Anaerobic Metabolism in Chlamydomonas reinhardtii

    PubMed Central

    Olson, Adam C.; Carter, Clay J.

    2016-01-01

    The unicellular green algae Chlamydomonas reinhardtii has long been studied for its unique fermentation pathways and has been evaluated as a candidate organism for biofuel production. Fermentation in C. reinhardtii is facilitated by a network of three predominant pathways producing four major byproducts: formate, ethanol, acetate and hydrogen. Previous microarray studies identified many genes as being highly up-regulated during anaerobiosis. For example, hybrid cluster protein 4 (HCP4) was found to be one of the most highly up-regulated genes under anoxic conditions. Hybrid cluster proteins have long been studied for their unique spectroscopic properties, yet their biological functions remain largely unclear. To probe its role during anaerobiosis, HCP4 was silenced using artificial microRNAs (ami-hcp4) followed by extensive phenotypic analyses of cells grown under anoxic conditions. Both the expression of key fermentative enzymes and their respective metabolites were significantly altered in ami-hcp4, with nitrogen uptake from the media also being significantly different than wild-type cells. The results strongly suggest a role for HCP4 in regulating key fermentative and nitrogen utilization pathways. PMID:26930496

  18. Oxidative stress contributes to autophagy induction in response to endoplasmic reticulum stress in Chlamydomonas reinhardtii.

    PubMed

    Pérez-Martín, Marta; Pérez-Pérez, María Esther; Lemaire, Stéphane D; Crespo, José L

    2014-10-01

    The accumulation of unfolded/misfolded proteins in the endoplasmic reticulum (ER) results in the activation of stress responses, such as the unfolded protein response or the catabolic process of autophagy to ultimately recover cellular homeostasis. ER stress also promotes the production of reactive oxygen species, which play an important role in autophagy regulation. However, it remains unknown whether reactive oxygen species are involved in ER stress-induced autophagy. In this study, we provide evidence connecting redox imbalance caused by ER stress and autophagy activation in the model unicellular green alga Chlamydomonas reinhardtii. Treatment of C. reinhardtii cells with the ER stressors tunicamycin or dithiothreitol resulted in up-regulation of the expression of genes encoding ER resident endoplasmic reticulum oxidoreductin1 oxidoreductase and protein disulfide isomerases. ER stress also triggered autophagy in C. reinhardtii based on the protein abundance, lipidation, cellular distribution, and mRNA levels of the autophagy marker ATG8. Moreover, increases in the oxidation of the glutathione pool and the expression of oxidative stress-related genes were detected in tunicamycin-treated cells. Our results revealed that the antioxidant glutathione partially suppressed ER stress-induced autophagy and decreased the toxicity of tunicamycin, suggesting that oxidative stress participates in the control of autophagy in response to ER stress in C. reinhardtii In close agreement, we also found that autophagy activation by tunicamycin was more pronounced in the C. reinhardtii sor1 mutant, which shows increased expression of oxidative stress-related genes.

  19. Production of biodiesel from microalgae Chlamydomonas polypyrenoideum grown on dairy industry wastewater.

    PubMed

    Kothari, Richa; Prasad, Ravindra; Kumar, Virendra; Singh, D P

    2013-09-01

    This study involves a process of phyco-remediation of dairy industry wastewater by algal strain Chlamydomonas polypyrenoideum. The results of selected algal strain indicated that dairy industry wastewater was good nutrient supplement for algal growth in comparable with BG-11 growth medium. Alga grown on dairy industry wastewater reduced the pollution load of nitrate (90%), nitrite (74%), phosphate (70%), chloride (61%), fluoride (58%), and ammonia (90%) on 10th day of its growth as compared to that of uninoculated wastewater. The lipid content of algal biomass grown on dairy wastewater on 10th day (1.6g) and 15th day (1.2 g) of batch experiment was found to be higher than the lipid content of algal biomass grown in BG-11 growth medium on 10th day (1.27 g) and 15th day (1.0 g) of batch experiment. The results on FTIR analysis of the extracted bio-oil through transesterification reaction was comparable with bio-oil obtained from other sources.

  20. Light/electricity conversion by defined cocultures of Chlamydomonas and Geobacter.

    PubMed

    Nishio, Koichi; Hashimoto, Kazuhito; Watanabe, Kazuya

    2013-04-01

    Biological energy-conversion systems are attractive in terms of their self-organizing and self-sustaining properties and are expected to be applied towards environmentally friendly bioenergy processes. Recent studies have demonstrated that sustainable light/electricity-conversion systems, termed microbial solar cells (MSCs), can be constructed using naturally occurring microbial communities. To better understand the energy-conversion mechanisms in microbial communities, the present study attempted to construct model MSCs comprised of defined cocultures of a green alga, Chlamydomonas reinhardtii, and an iron-reducing bacterium, Geobacter sulfurreducens, and examined their metabolism and interactions in MSCs. When MSC bioreactors were inoculated with these microbes and irradiated on a 12-h light/dark cycle, periodic current was generated in the dark with energy-conversion efficiencies of 0.1%. Metabolite analyses revealed that G. sulfurreducens generated current by oxidizing formate that was produced by C. reinhardtii in the dark. These results demonstrate that the light/electricity conversion occurs via syntrophic interactions between phototrophs and electricity-generating bacteria. Based on the results and data in literatures, it is estimated that the excretion of organics by the phototroph was the bottleneck step in the syntrophic light/electricity conversion. We also discuss differences between natural-community and defined-coculture MSCs.

  1. Dynamic Interplay between Nucleoid Segregation and Genome Integrity in Chlamydomonas Chloroplasts1[OPEN

    PubMed Central

    Odahara, Masaki; Kobayashi, Yusuke; Shikanai, Toshiharu; Nishimura, Yoshiki

    2016-01-01

    The chloroplast (cp) genome is organized as nucleoids that are dispersed throughout the cp stroma. Previously, a cp homolog of bacterial recombinase RecA (cpRECA) was shown to be involved in the maintenance of cp genome integrity by repairing damaged chloroplast DNA and by suppressing aberrant recombination between short dispersed repeats in the moss Physcomitrella patens. Here, overexpression and knockdown analysis of cpRECA in the green alga Chlamydomonas reinhardtii revealed that cpRECA was involved in cp nucleoid dynamics as well as having a role in maintaining cp genome integrity. Overexpression of cpRECA tagged with yellow fluorescent protein or hemagglutinin resulted in the formation of giant filamentous structures that colocalized exclusively to chloroplast DNA and cpRECA localized to cp nucleoids in a heterogenous manner. Knockdown of cpRECA led to a significant reduction in cp nucleoid number that was accompanied by nucleoid enlargement. This phenotype resembled those of gyrase inhibitor-treated cells and monokaryotic chloroplast mutant cells and suggested that cpRECA was involved in organizing cp nucleoid dynamics. The cp genome also was destabilized by induced recombination between short dispersed repeats in cpRECA-knockdown cells and gyrase inhibitor-treated cells. Taken together, these results suggest that cpRECA and gyrase are both involved in nucleoid dynamics and the maintenance of genome integrity and that the mechanisms underlying these processes may be intimately related in C. reinhardtii cps. PMID:27756821

  2. Improved automated monitoring and new analysis algorithm for circadian phototaxis rhythms in Chlamydomonas

    PubMed Central

    Gaskill, Christa; Forbes-Stovall, Jennifer; Kessler, Bruce; Young, Mike; Rinehart, Claire A.; Jacobshagen, Sigrid

    2010-01-01

    Automated monitoring of circadian rhythms is an efficient way of gaining insight into oscillation parameters like period and phase for the underlying pacemaker of the circadian clock. Measurement of the circadian rhythm of phototaxis (swimming towards light) exhibited by the green alga Chlamydomonas reinhardtii has been automated by directing a narrow and dim light beam through a culture at regular intervals and determining the decrease in light transmittance due to the accumulation of cells in the beam. In this study, the monitoring process was optimized by constructing a new computer-controlled measuring machine that limits the test beam to wavelengths reported to be specific for phototaxis and by choosing an algal strain, which does not need background illumination between test light cycles for proper expression of the rhythm. As a result, period and phase of the rhythm are now unaffected by the time a culture is placed into the machine. Analysis of the rhythm data was also optimized through a new algorithm, whose robustness was demonstrated using virtual rhythms with various noises. The algorithm differs in particular from other reported algorithms by maximizing the fit of the data to a sinusoidal curve that dampens exponentially. The algorithm was also used to confirm the reproducibility of rhythm monitoring by the machine. Machine and algorithm can now be used for a multitude of circadian clock studies that require unambiguous period and phase determinations such as light pulse experiments to identify the photoreceptor(s) that reset the circadian clock in C. reinhardtii. PMID:20116270

  3. Comparative study of phototactic and photophobic receptor chromophore properties in Chlamydomonas reinhardtii.

    PubMed Central

    Zacks, D N; Derguini, F; Nakanishi, K; Spudich, J L

    1993-01-01

    The motile, unicellular, eukaryotic alga Chlamydomonas reinhardtii exhibits two distinct behavioral reactions to light stimuli, phototaxis and the photophobic response. Both are mediated by retinal-containing receptors. This paper focuses on a direct comparison of the two photoresponses and the chromophore requirements for their photoreceptor(s). Using computerized motion analysis assays for phototaxis and photophobic responses by the same populations of cells, we measured the ability of various isomers and analogues of retinal to reconstitute photobehavior in the pigment-deficient mutant FN68. The results indicate that photophobic and phototaxis responses each require chromophores with an all-trans polyene chain configuration, planar ionone ring/polyene chain conformation, and the ability to isomerize around the retinal C13-C14 double bond. One difference between the two behaviors is that the photophobic response becomes highly desensitized after light stimuli to which the phototaxis response does not become desensitized, indicating the existence of at least one distinct step in the photophobic response pathway. A second difference is that the retinal regeneration of the photophobic response but not of phototaxis is inhibited by a 5-membered ring 13-trans-locked analogue. While showing close similarity in the chromophore structural requirements of the two behaviors, the results indicate that differences exist between the two responses at the level of their photoreceptor proteins and/or in their transduction processes. Images FIGURE 2 PMID:8369455

  4. Fitness effects of new mutations in Chlamydomonas reinhardtii across two stress gradients.

    PubMed

    Kraemer, S A; Morgan, A D; Ness, R W; Keightley, P D; Colegrave, N

    2016-03-01

    Most spontaneous mutations affecting fitness are likely to be deleterious, but the strength of selection acting on them might be impacted by environmental stress. Such stress-dependent selection could expose hidden genetic variation, which in turn might increase the adaptive potential of stressed populations. On the other hand, this variation might represent a genetic load and thus lead to population extinction under stress. Previous studies to determine the link between stress and mutational effects on fitness, however, have produced inconsistent results. Here, we determined the net change in fitness in 29 genotypes of the green algae Chlamydomonas reinhardtii that accumulated mutations in the near absence of selection for approximately 1000 generations across two stress gradients, increasing NaCl and decreasing phosphate. We found mutational effects to be magnified under extremely stressful conditions, but such effects were specific both to the type of stress and to the genetic background. The detection of stress-dependent fitness effects of mutations depended on accurately scaling relative fitness measures by generation times, thus offering an explanation for the inconsistencies among previous studies.

  5. Characterization of the EYE2 gene required for eyespot assembly in Chlamydomonas reinhardtii.

    PubMed Central

    Roberts, D G; Lamb, M R; Dieckmann, C L

    2001-01-01

    The unicellular biflagellate green alga Chlamydomonas reinhardtii can perceive light and respond by altering its swimming behavior. The eyespot is a specialized structure for sensing light, which is assembled de novo at every cell division from components located in two different cellular compartments. Photoreceptors and associated signal transduction components are localized in a discrete patch of the plasma membrane. This patch is tightly packed against an underlying sandwich of chloroplast membranes and carotenoid-filled lipid granules, which aids the cell in distinguishing light direction. In a prior screen for mutant strains with eyespot defects, the EYE2 locus was defined by the single eye2-1 allele. The mutant strain has no eyespot by light microscopy and has no organized carotenoid granule layers as judged by electron microscopy. Here we demonstrate that the eye2-1 mutant is capable of responding to light, although the strain is far less sensitive than wild type to low light intensities and orients imprecisely. Therefore, pigment granule layer assembly in the chloroplast is not required for photoreceptor localization in the plasma membrane. A plasmid-insertion mutagenesis screen yielded the eye2-2 allele, which allowed the isolation and characterization of the EYE2 gene. The EYE2 protein is a member of the thioredoxin superfamily. Site-directed mutagenesis of the active site cysteines demonstrated that EYE2 function in eyespot assembly is redox independent, similar to the auxiliary functions of other thioredoxin family members in protein folding and complex assembly. PMID:11454753

  6. Evolution of salt tolerance in a laboratory reared population of Chlamydomonas reinhardtii.

    PubMed

    Perrineau, Marie-Mathilde; Zelzion, Ehud; Gross, Jeferson; Price, Dana C; Boyd, Jeffrey; Bhattacharya, Debashish

    2014-06-01

    Understanding the genetic underpinnings of adaptive traits in microalgae is important for the study of evolution and for applied uses. We used long-term selection under a regime of serial transfers with haploid populations of the green alga Chlamydomonas reinhardtii raised in liquid TAP medium containing 200 mM NaCl. After 1255 generations, evolved salt (ES) populations could grow as rapidly in high salt medium as progenitor cells (progenitor light [PL]). Transcriptome data were analysed to elucidate the basis of salt tolerance in ES cells when compared with PL cells and to cells incubated for 48 h in high salt medium (progenitor salt [PS], the short-term acclimation response). These data demonstrate that evolved and short-term acclimation responses to salt stress differ fundamentally from each other. Progenitor salt cells exhibit well-known responses to salt stress such as reduction in photosynthesis, upregulation of glycerophospholipid signaling, and upregulation of the transcription and translation machinery. In contrast, ES cells show downregulation of genes involved in the stress response and in transcription/translation. Our results suggest that gene-rich mixotrophic lineages such as C. reinhardtii may be able to adapt rapidly to abiotic stress engendered either by a rapidly changing climate or physical vicariance events that isolate populations in stressful environments.

  7. Glutathionylation in the photosynthetic model organism Chlamydomonas reinhardtii: a proteomic survey.

    PubMed

    Zaffagnini, Mirko; Bedhomme, Mariette; Groni, Hayam; Marchand, Christophe H; Puppo, Carine; Gontero, Brigitte; Cassier-Chauvat, Corinne; Decottignies, Paulette; Lemaire, Stéphane D

    2012-02-01

    Protein glutathionylation is a redox post-translational modification occurring under oxidative stress conditions and playing a major role in cell regulation and signaling. This modification has been mainly studied in nonphotosynthetic organisms, whereas much less is known in photosynthetic organisms despite their important exposure to oxidative stress caused by changes in environmental conditions. We report a large scale proteomic analysis using biotinylated glutathione and streptavidin affinity chromatography that allowed identification of 225 glutathionylated proteins in the eukaryotic unicellular green alga Chlamydomonas reinhardtii. Moreover, 56 sites of glutathionylation were also identified after peptide affinity purification and tandem mass spectrometry. The targets identified belong to a wide range of biological processes and pathways, among which the Calvin-Benson cycle appears to be a major target. The glutathionylation of four enzymes of this cycle, phosphoribulokinase, glyceraldehyde-3-phosphate dehydrogenase, ribose-5-phosphate isomerase, and phosphoglycerate kinase was confirmed by Western blot and activity measurements. The results suggest that glutathionylation could constitute a major mechanism of regulation of the Calvin-Benson cycle under oxidative stress conditions.

  8. The Involvement of hybrid cluster protein 4, HCP4, in Anaerobic Metabolism in Chlamydomonas reinhardtii.

    PubMed

    Olson, Adam C; Carter, Clay J

    2016-01-01

    The unicellular green algae Chlamydomonas reinhardtii has long been studied for its unique fermentation pathways and has been evaluated as a candidate organism for biofuel production. Fermentation in C. reinhardtii is facilitated by a network of three predominant pathways producing four major byproducts: formate, ethanol, acetate and hydrogen. Previous microarray studies identified many genes as being highly up-regulated during anaerobiosis. For example, hybrid cluster protein 4 (HCP4) was found to be one of the most highly up-regulated genes under anoxic conditions. Hybrid cluster proteins have long been studied for their unique spectroscopic properties, yet their biological functions remain largely unclear. To probe its role during anaerobiosis, HCP4 was silenced using artificial microRNAs (ami-hcp4) followed by extensive phenotypic analyses of cells grown under anoxic conditions. Both the expression of key fermentative enzymes and their respective metabolites were significantly altered in ami-hcp4, with nitrogen uptake from the media also being significantly different than wild-type cells. The results strongly suggest a role for HCP4 in regulating key fermentative and nitrogen utilization pathways.

  9. Isolation of Chlamydomonas reinhardtii mutants with altered mitochondrial respiration by chlorophyll fluorescence measurement.

    PubMed

    Massoz, Simon; Larosa, Véronique; Horrion, Bastien; Matagne, René F; Remacle, Claire; Cardol, Pierre

    2015-12-10

    The unicellular green alga Chlamydomonas reinhardtii is a model organism for studying energetic metabolism. Most mitochondrial respiratory-deficient mutants characterized to date have been isolated on the basis of their reduced ability to grow in heterotrophic conditions. Mitochondrial deficiencies are usually partly compensated by adjustment of photosynthetic activity and more particularly by transition to state 2. In this work, we explored the opportunity to select mutants impaired in respiration and/or altered in dark metabolism by measuring maximum photosynthetic efficiency by chlorophyll fluorescence analyses (FV/FM). Out of about 2900 hygromycin-resistant insertional mutants generated from wild type or from a mutant strain deficient in state transitions (stt7 strain), 22 were found to grow slowly in heterotrophic conditions and 8 of them also showed a lower FV/FM value. Several disrupted coding sequences were identified, including genes coding for three different subunits of respiratory-chain complex I (NUO9, NUOA9, NUOP4) or for isocitrate lyase (ICL1). Overall, the comparison of respiratory mutants obtained in wild-type or stt7 genetic backgrounds indicated that the FV/FM value can be used to isolate mutants severely impaired in dark metabolism.

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

    SciTech Connect

    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.

  11. A flavin binding cryptochrome photoreceptor responds to both blue and red light in Chlamydomonas reinhardtii.

    PubMed

    Beel, Benedikt; Prager, Katja; Spexard, Meike; Sasso, Severin; Weiss, Daniel; Müller, Nico; Heinnickel, Mark; Dewez, David; Ikoma, Danielle; Grossman, Arthur R; Kottke, Tilman; Mittag, Maria

    2012-07-01

    Cryptochromes are flavoproteins that act as sensory blue light receptors in insects, plants, fungi, and bacteria. We have investigated a cryptochrome from the green alga Chlamydomonas reinhardtii with sequence homology to animal cryptochromes and (6-4) photolyases. In response to blue and red light exposure, this animal-like cryptochrome (aCRY) alters the light-dependent expression of various genes encoding proteins involved in chlorophyll and carotenoid biosynthesis, light-harvesting complexes, nitrogen metabolism, cell cycle control, and the circadian clock. Additionally, exposure to yellow but not far-red light leads to comparable increases in the expression of specific genes; this expression is significantly reduced in an acry insertional mutant. These in vivo effects are congruent with in vitro data showing that blue, yellow, and red light, but not far-red light, are absorbed by the neutral radical state of flavin in aCRY. The aCRY neutral radical is formed following blue light absorption of the oxidized flavin. Red illumination leads to conversion to the fully reduced state. Our data suggest that aCRY is a functionally important blue and red light-activated flavoprotein. The broad spectral response implies that the neutral radical state functions as a dark form in aCRY and expands the paradigm of flavoproteins and cryptochromes as blue light sensors to include other light qualities.

  12. Regulation of Oil Biosynthesis in Algae

    DTIC Science & Technology

    2011-03-14

    transportation fuels can potentially be addressed by exploring oil (triacylglycerol) biosynthesis in microalgae . Many microalgae , including Chlamydomonas...biosynthesis in microalgae have not been studied at the molecular level. Chlamydomonas is being used as a microalgal model to identify genes and regulatory...of this phenomenon will shed light on the physiological significance of oil production in microalgae . A first paper describing this interesting

  13. Phenotypic diversity of hydrogen production in chlorophycean algae reflects distinct anaerobic metabolisms.

    PubMed

    Meuser, Jonathan E; Ananyev, Gennady; Wittig, Lauren E; Kosourov, Sergey; Ghirardi, Maria L; Seibert, Michael; Dismukes, G Charles; Posewitz, Matthew C

    2009-06-01

    Several species of green algae use [FeFe]-hydrogenases to oxidize and/or produce H(2) during anoxia. To further define unique aspects of algal hydrogenase activity, the well-studied anaerobic metabolisms of Chlamydomonas reinhardtii were compared with four strains of Chlamydomonas moewusii and a Lobochlamys culleus strain. In vivo and in vitro hydrogenase activity, starch accumulation/degradation, and anaerobic end product secretion were analyzed. The C. moewusii strains showed the most rapid induction of hydrogenase activity, congruent with high rates of starch catabolism, and anoxic metabolite accumulation. Intriguingly, we observed significant differences in morphology and hydrogenase activity in the C. moewusii strains examined, likely the result of long-term adaptation and/or genetic drift during culture maintenance. Of the C. moewusii strains examined, SAG 24.91 showed the highest in vitro hydrogenase activity. However, SAG 24.91 produced little H(2) under conditions of sulfur limitation, which is likely a consequence of its inability to utilize exogenous acetate. In L. culleus, hydrogenase activity was minimal unless pulsed light was used to induce significant H(2) photoproduction. Overall, our results demonstrate that unique anaerobic acclimation strategies have evolved in distinct green algae, resulting in differential levels of hydrogenase activity and species-specific patterns of NADH reoxidation during anoxia.

  14. Phenotypic Diversity of Hydrogen Production in Chlorophycean Algae Reflects Distinct Anaerobic Metabolisms

    SciTech Connect

    Meuser, J. E.; Ananyev, G.; Wittig, L. E.; Kosourov, S.; Ghirardi, M. L.; Seibert, M.; Dismukes, G. C.; Posewitz, M. C.

    2009-01-01

    Several species of green algae use [FeFe]-hydrogenases to oxidize and/or produce H{sub 2} during anoxia. To further define unique aspects of algal hydrogenase activity, the well-studied anaerobic metabolisms of Chlamydomonas reinhardtii were compared with four strains of Chlamydomonas moewusii and a Lobochlamys culleus strain. In vivo and in vitro hydrogenase activity, starch accumulation/degradation, and anaerobic end product secretion were analyzed. The C. moewusii strains showed the most rapid induction of hydrogenase activity, congruent with high rates of starch catabolism, and anoxic metabolite accumulation. Intriguingly, we observed significant differences in morphology and hydrogenase activity in the C. moewusii strains examined, likely the result of long-term adaptation and/or genetic drift during culture maintenance. Of the C. moewusii strains examined, SAG 24.91 showed the highest in vitro hydrogenase activity. However, SAG 24.91 produced little H{sub 2} under conditions of sulfur limitation, which is likely a consequence of its inability to utilize exogenous acetate. In L. culleus, hydrogenase activity was minimal unless pulsed light was used to induce significant H2 photoproduction. Overall, our results demonstrate that unique anaerobic acclimation strategies have evolved in distinct green algae, resulting in differential levels of hydrogenase activity and species-specific patterns of NADH reoxidation during anoxia.

  15. Solar-driven hydrogen production in green algae.

    PubMed

    Burgess, Steven J; Tamburic, Bojan; Zemichael, Fessehaye; Hellgardt, Klaus; Nixon, Peter J

    2011-01-01

    The twin problems of energy security and global warming make hydrogen an attractive alternative to traditional fossil fuels with its combustion resulting only in the release of water vapor. Biological hydrogen production represents a renewable source of the gas and can be performed by a diverse range of microorganisms from strict anaerobic bacteria to eukaryotic green algae. Compared to conventional methods for generating H(2), biological systems can operate at ambient temperatures and pressures without the need for rare metals and could potentially be coupled to a variety of biotechnological processes ranging from desalination and waste water treatment to pharmaceutical production. Photobiological hydrogen production by microalgae is particularly attractive as the main inputs for the process (water and solar energy) are plentiful. This chapter focuses on recent developments in solar-driven H(2) production in green algae with emphasis on the model organism Chlamydomonas reinhardtii. We review the current methods used to achieve sustained H(2) evolution and discuss possible approaches to improve H(2) yields, including the optimization of culturing conditions, reducing light-harvesting antennae and targeting auxiliary electron transport and fermentative pathways that compete with the hydrogenase for reductant. Finally, industrial scale-up is discussed in the context of photobioreactor design and the future prospects of the field are considered within the broader context of a biorefinery concept.

  16. [Ecological characteristic of benthic epipelic algae and the characteristic of water environment quality in heavily polluted river in city].

    PubMed

    Zhao, Zhen-hua; Ruan, Xiao-hong; Xing, Ya-nan; Ni, Li-xiao; Gao, Li-cun

    2009-12-01

    The water quality and algae community of Nanyuan Water System in the old city area of Suzhou were monitored for a year. Results showed that the water pollution in the studied area was mainly related to nitrogen (NH4+ -N and TN). Sometimes, they even exceeded the Environmental Quality Standards for Surface Water (GB 3838-2002, PRC) more than 5 times. 34 species of benthic epipelic algae were observed by microscope, and the species amount of diatom algae, green algae and blue algae are more than others. Their abundance and biomass are far higher than that of the pelagic algae in the same sites,and reach 2 145.5 x 10(4) cells/mL and 3.524 mg/mL,respectively. The dominant species of benthic epipelic algae in Nanyuan's water system are diatom algae and blue algae, most of which belong to the heterotrophic type or bi-trophic type algae, the typical genera include: Oscillaria amphibian (affiliated to Cyanophyta), Cyclotella sp., Melosira sp., Stephanodiscus hantzschii, Navicula sp., Nitzschia sp., Gomphonema (affiliated to Bacillariophyta) and so on. And their distribution of species and abundance are very nonuniform in different reach of heavily polluted city river, which relates to the pollutant characteristics of the river. The seasonal variety trend of the abundance for benthic algae showed that:summer > autumn > spring > winter, and that of biomass for benthic algae showed that: the biomass in winter is the most of four seasons and change extent of the biomass is not obvious in spring, summer and autumn. The research results can provide reference for the ecology restoration of city heavily polluted river.

  17. Screening for bioactive compounds from algae.

    PubMed

    Plaza, M; Santoyo, S; Jaime, L; García-Blairsy Reina, G; Herrero, M; Señoráns, F J; Ibáñez, E

    2010-01-20

    In the present work, a comprehensive methodology to carry out the screening for novel natural functional compounds is presented. To do that, a new strategy has been developed including the use of unexplored natural sources (i.e., algae and microalgae) together with environmentally clean extraction techniques and advanced analytical tools. The developed procedure allows also estimating the functional activities of the different extracts obtained and even more important, to correlate these activities with their particular chemical composition. By applying this methodology it has been possible to carry out the screening for bioactive compounds in the algae Himanthalia elongata and the microalgae Synechocystis sp. Both algae produced active extracts in terms of both antioxidant and antimicrobial activity. The obtained pressurized liquid extracts were chemically characterized by GC-MS and HPLC-DAD. Different fatty acids and volatile compounds with antimicrobial activity were identified, such as phytol, fucosterol, neophytadiene or palmitic, palmitoleic and oleic acids. Based on the results obtained, ethanol was selected as the most appropriate solvent to extract this kind of compounds from the natural sources studied.

  18. Genome-wide long non-coding RNA screening, identification and characterization in a model microorganism Chlamydomonas reinhardtii.

    PubMed

    Li, Hui; Wang, Yuting; Chen, Meirong; Xiao, Peng; Hu, Changxing; Zeng, Zhiyong; Wang, Chaogang; Wang, Jiangxin; Hu, Zhangli

    2016-09-23

    Microalgae are regarded as the most promising biofuel candidates and extensive metabolic engineering were conducted but very few improvements were achieved. Long non-coding RNA (lncRNA) investigation and manipulation may provide new insights for this issue. LncRNAs refer to transcripts that are longer than 200 nucleotides, do not encode proteins but play important roles in eukaryotic gene regulation. However, no information of potential lncRNAs has been reported in eukaryotic alga. Recently, we performed RNA sequencing in Chlamydomonas reinhardtii, and obtained totally 3,574 putative lncRNAs. 1440 were considered as high-confidence lncRNAs, including 936 large intergenic, 310 intronic and 194 anti-sense lncRNAs. The average transcript length, ORF length and numbers of exons for lncRNAs are much less than for genes in this green alga. In contrast with human lncRNAs of which more than 98% are spliced, the percentage in C. reinhardtii is only 48.1%. In addition, we identified 367 lncRNAs responsive to sulfur deprivation, including 36 photosynthesis-related lncRNAs. This is the first time that lncRNAs were explored in the unicellular model organism C. reinhardtii. The lncRNA data could also provide new insights into C. reinhardtii hydrogen production under sulfur deprivation.

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

    PubMed Central

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

    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 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. PMID:27335457

  20. Genome-wide long non-coding RNA screening, identification and characterization in a model microorganism Chlamydomonas reinhardtii

    PubMed Central

    Li, Hui; Wang, Yuting; Chen, Meirong; Xiao, Peng; Hu, Changxing; Zeng, Zhiyong; Wang, Chaogang; Wang, Jiangxin; Hu, Zhangli

    2016-01-01

    Microalgae are regarded as the most promising biofuel candidates and extensive metabolic engineering were conducted but very few improvements were achieved. Long non-coding RNA (lncRNA) investigation and manipulation may provide new insights for this issue. LncRNAs refer to transcripts that are longer than 200 nucleotides, do not encode proteins but play important roles in eukaryotic gene regulation. However, no information of potential lncRNAs has been reported in eukaryotic alga. Recently, we performed RNA sequencing in Chlamydomonas reinhardtii, and obtained totally 3,574 putative lncRNAs. 1440 were considered as high-confidence lncRNAs, including 936 large intergenic, 310 intronic and 194 anti-sense lncRNAs. The average transcript length, ORF length and numbers of exons for lncRNAs are much less than for genes in this green alga. In contrast with human lncRNAs of which more than 98% are spliced, the percentage in C. reinhardtii is only 48.1%. In addition, we identified 367 lncRNAs responsive to sulfur deprivation, including 36 photosynthesis-related lncRNAs. This is the first time that lncRNAs were explored in the unicellular model organism C. reinhardtii. The lncRNA data could also provide new insights into C. reinhardtii hydrogen production under sulfur deprivation. PMID:27659799

  1. A Transient Receptor Potential Ion Channel in Chlamydomonas Shares Key Features with Sensory Transduction-Associated TRP Channels in Mammals

    PubMed Central

    Arias-Darraz, Luis; Cabezas, Deny; Colenso, Charlotte K.; Alegría-Arcos, Melissa; Bravo-Moraga, Felipe; Varas-Concha, Ignacio; Almonacid, Daniel E.; Madrid, Rodolfo; Brauchi, Sebastian

    2015-01-01

    Sensory modalities are essential for navigating through an ever-changing environment. From insects to mammals, transient receptor potential (TRP) channels are known mediators for cellular sensing. Chlamydomonas reinhardtii is a motile single-celled freshwater green alga that is guided by photosensory, mechanosensory, and chemosensory cues. In this type of alga, sensory input is first detected by membrane receptors located in the cell body and then transduced to the beating cilia by membrane depolarization. Although TRP channels seem to be absent in plants, C. reinhardtii possesses genomic sequences encoding TRP proteins. Here, we describe the cloning and characterization of a C. reinhardtii version of a TRP channel sharing key features present in mammalian TRP channels associated with sensory transduction. In silico sequence-structure analysis unveiled the modular design of TRP channels, and electrophysiological experiments conducted on Human Embryonic Kidney-293T cells expressing the Cr-TRP1 clone showed that many of the core functional features of metazoan TRP channels are present in Cr-TRP1, suggesting that basic TRP channel gating characteristics evolved early in the history of eukaryotes. PMID:25595824

  2. Transcriptional response to copper excess and identification of genes involved in heavy metal tolerance in the extremophilic microalga Chlamydomonas acidophila.

    PubMed

    Olsson, Sanna; Puente-Sánchez, Fernando; Gómez, Manuel J; Aguilera, Angeles

    2015-05-01

    High concentrations of heavy metals are typical of acidic environments. Therefore, studies on acidophilic organisms in their natural environments improve our understanding on the evolution of heavy metal tolerance and detoxification in plants. Here we sequenced the transcriptome of the extremophilic microalga Chlamydomonas acidophila cultivated in control conditions and with 500 μM of copper for 24 h. High-throughput 454 sequencing was followed by de novo transcriptome assembly. The reference transcriptome was annotated and genes related to heavy metal tolerance and abiotic stress were identified. Analyses of differentially expressed transcripts were used to detect genes involved in metabolic pathways related to abiotic stress tolerance, focusing on effects caused by increased levels of copper. Both transcriptomic data and observations from PAM fluorometry analysis suggested that the photosynthetic activity of C. acidophila is not adversely affected by addition of high amounts of copper. Up-regulated transcripts include several transcripts related to photosynthesis and carbohydrate metabolism, transcripts coding for general stress response, and a transcript annotated as homologous to the oil-body-associated protein HOGP coding gene. The first de novo assembly of C. acidophila significantly increases transcriptomic data available on extremophiles and green algae and thus provides an important reference for further molecular genetic studies. The differences between differentially expressed transcripts detected in our study suggest that the response to heavy metal exposure in C. acidophila is different from other studied green algae.

  3. Stress responses and metal tolerance of Chlamydomonas acidophila in metal-enriched lake water and artificial medium.

    PubMed

    Spijkerman, Elly; Barua, Deepak; Gerloff-Elias, Antje; Kern, Jürgen; Gaedke, Ursula; Heckathorn, Scott A

    2007-07-01

    Chlamydomonas acidophila faces high heavy-metal concentrations in acidic mining lakes, where it is a dominant phytoplankton species. To investigate the importance of metals to C. acidophila in these lakes, we examined the response of growth, photosynthesis, cell structure, heat-shock protein (Hsp) accumulation, and metal adsorption after incubation in metal-rich lake water and artificial growth medium enriched with metals (Fe, Zn). Incubation in both metal-rich lake water and medium caused large decreases in photosystem II function (though no differences among lakes), but no decrease in growth rate (except for medium + Fe). Concentrations of small Hsps were higher in algae incubated in metal-rich lake-water than in metal-enriched medium, whereas Hsp60 and Hsp70A were either less or equally expressed. Cellular Zn and Fe contents were lower, and metals adsorbed to the cell surface were higher, in lake-water-incubated algae than in medium-grown cells. The results indicate that high Zn or Fe levels are likely not the main or only contributor to the low primary production in mining lakes, and multiple adaptations of C. acidophila (e.g., high Hsp levels, decreased metal accumulation) increase its tolerance to metals and permit survival under such adverse environmental conditions. Supposedly, the main stress factor present in the lake water is an interaction between low P and high Fe concentrations.

  4. Proteomic Analysis of the Eyespot of Chlamydomonas reinhardtii Provides Novel Insights into Its Components and Tactic Movements[W

    PubMed Central

    Schmidt, Melanie; Geßner, Gunther; Luff, Matthias; Heiland, Ines; Wagner, Volker; Kaminski, Marc; Geimer, Stefan; Eitzinger, Nicole; Reißenweber, Tobias; Voytsekh, Olga; Fiedler, Monika; Mittag, Maria; Kreimer, Georg

    2006-01-01

    Flagellate green algae have developed a visual system, the eyespot apparatus, which allows the cell to phototax. To further understand the molecular organization of the eyespot apparatus and the phototactic movement that is controlled by light and the circadian clock, a detailed understanding of all components of the eyespot apparatus is needed. We developed a procedure to purify the eyespot apparatus from the green model alga Chlamydomonas reinhardtii. Its proteomic analysis resulted in the identification of 202 different proteins with at least two different peptides (984 in total). These data provide new insights into structural components of the eyespot apparatus, photoreceptors, retina(l)-related proteins, members of putative signaling pathways for phototaxis and chemotaxis, and metabolic pathways within an algal visual system. In addition, we have performed a functional analysis of one of the identified putative components of the phototactic signaling pathway, casein kinase 1 (CK1). CK1 is also present in the flagella and thus is a promising candidate for controlling behavioral responses to light. We demonstrate that silencing CK1 by RNA interference reduces its level in both flagella and eyespot. In addition, we show that silencing of CK1 results in severe disturbances in hatching, flagellum formation, and circadian control of phototaxis. PMID:16798888

  5. Omics in Chlamydomonas for Biofuel Production.

    PubMed

    Aucoin, Hanna R; Gardner, Joseph; Boyle, Nanette R

    2016-01-01

    In response to demands for sustainable domestic fuel sources, research into biofuels has become increasingly important. Many challenges face biofuels in their effort to replace petroleum fuels, but rational strain engineering of algae and photosynthetic organisms offers a great deal of promise. For decades, mutations and stress responses in photosynthetic microbiota were seen to result in production of exciting high-energy fuel molecules, giving hope but minor capability for design. However, '-omics' techniques for visualizing entire cell processing has clarified biosynthesis and regulatory networks. Investigation into the promising production behaviors of the model organism C. reinhardtii and its mutants with these powerful techniques has improved predictability and understanding of the diverse, complex interactions within photosynthetic organisms. This new equipment has created an exciting new frontier for high-throughput, predictable engineering of photosynthetically produced carbon-neutral biofuels.

  6. SOME FACTORS IN THE COMPETITION OR ANTAGONISM AMONG BACTERIA, ALGAE, AND AQUATIC WEEDS.

    PubMed

    Filzgerald, G P

    1969-12-01

    Field observations of changes in the populations of aquatic weeds and phytoplankton have confirmed that aquatic weeds have antagonistic activity toward phytoplankton. Nutritional studies in the laboratory indicate that cultures of the aquatic weeds, Myriophyllum sp., Ceratophyllum sp., and duckweed (Lemma minor L.); liquid cultures of barley (Hordeum vulgare L., Dickson variety); and cultures of the filamentous green algae, Cladophora sp. and Pithophora oedogonium (Mont.) Withrock, will remain relatively free of epiphytes or competing phytoplankton if the cultures are nitrogen-limited. Field observations of Cladophora sp. have confirmed that the growth of epiphytes on the Cladophora is related to conditions of surplus available nitrogen compounds. It is proposed that this antagonistic activity may be due to a "nitrogen sink" effect in which the aquatic weeds or filamentous green algae prevent the growth of contaminating algae by competition for the limited nitrogen compounds available. However, the presence of bacteria-sized organisms which have selective toxicity to certain algae indicates that perhaps multiple factors exist. Discussed are the ecological implications of associations of certain algae with bacteria that have selective toxicities for other species of algae under certain environmental conditions such as nitrogen-limited growth.

  7. Toxicity and bioaccumulation kinetics of arsenate in two freshwater green algae under different phosphate regimes.

    PubMed

    Wang, Ning-Xin; Li, Yan; Deng, Xi-Hai; Miao, Ai-Jun; Ji, Rong; Yang, Liu-Yan

    2013-05-01

    In the present study, the toxicity and bioaccumulation kinetics of arsenate in two green algae Chlamydomonas reinhardtii and Scenedesmus obliquus under phosphate-enriched (+P) and limited (-P) conditions were investigated. P-limitation was found to aggravate arsenate toxicity and S. obliquus was more tolerant than C. reinhardtii. Such phosphate-condition-dependent or algal-species-specific toxicity difference was narrowed when the relative inhibition of cell growth was plotted against intracellular arsenate content instead of its extracellular concentration. The discrepance was further reduced when the intracellular ratio of arsenic to phosphorus was applied. It suggests that both arsenate bioaccumulation and intracellular phosphorus played an important role in arsenate toxicity. On the other hand, arsenate uptake was induced by P-limitation and its variation with ambient arsenate concentration could be well fitted to the Michaelis-Menten model. Arsenate transporters of S. obliquus were found to have a higher affinity but lower capacity than those of C. reinhardtii, which explains its better regulation of arsenate accumulation than the latter species in the toxicity experiment. Further, arsenate depuration was facilitated and more was transformed to arsenite in C. reinhardtii or under -P condition. Intracellular proportion of arsenite was also increased after the algae were transferred from the long-term uptake media to a relatively clean solution in the efflux experiment. Both phenomena imply that algae especially the sensitive species could make physiological adjustments to alleviate the adverse effects of arsenate. Overall, our findings will facilitate the application of algae in arsenate remediation.

  8. Laccase-like enzyme activities from chlorophycean green algae with potential for bioconversion of phenolic pollutants.

    PubMed

    Otto, Benjamin; Beuchel, Carl; Liers, Christiane; Reisser, Werner; Harms, Hauke; Schlosser, Dietmar

    2015-06-01

    In order to explore the abundance and potential environmental functions of green algal laccases, we screened various algae for extracellular laccase-like activities, characterized basic features of these activities in selected species and exemplarily studied the transformation of environmental pollutants and complex natural compounds by the laccase of Tetracystis aeria. Oxidation of the classical laccase substrate ABTS was found to be widespread in chlorophycean algae. The oxidation activity detected in members of the 'Scenedesmus' clade was caused by an unknown thermostable low-molecular-mass compound. In contrast, species of the Moewusinia, including Chlamydomonas moewusii and T. aeria, excreted putative 'true' laccases. Phenolic substrates were oxidized by these enzymes optimally at neutral to alkaline pH. The Tetracystis laccase efficiently transformed bisphenol A, 17α-ethinylestradiol, nonylphenol and triclosan in the presence of ABTS as redox mediator, while anthracene, veratrylalcohol and adlerol were not attacked. Lignosulfonate and humic acid underwent slight (de)polymerization reactions in the presence of the laccase and mediator(s), probably involving the oxidation of phenolic constituents. Possible natural functions of the enzymes, such as the synthesis of complex polymers or detoxification processes, may assist the survival of the algae in adverse environments. In contaminated surface waters, laccase-producing green algae might contribute to the environmental breakdown of phenolic pollutants.

  9. Importance of algae as a potential source of biofuel.

    PubMed

    Singh, A K; Singh, M P

    2014-12-24

    Algae have a great potential source of biofuels and also have unique importance to reduce gaseous emissions, greenhouse gases, climatic changes, global warming receding of glaciers, rising sea levels and loss of biodiversity. The microalgae, like Scenedesmus obliquus, Neochloris oleabundans, Nannochloropsis sp., Chlorella emersonii, and Dunaliella tertiolecta have high oil content. Among the known algae, Scenedesmus obliquus is one of the most potential sources for biodiesel as it has adequate fatty acid (linolenic acid) and other polyunsaturated fatty acids. Bio—ethanol is already in the market of United States of America and Europe as an additive in gasoline. Bio—hydrogen is the cleanest biofuel and extensive efforts are going on to bring it to market at economical price. This review highlights recent development and progress in the field of algae as a potential source of biofuel.

  10. Chlamydomonas reinhardtii Secretes Compounds That Mimic Bacterial Signals and Interfere with Quorum Sensing Regulation in Bacteria1

    PubMed Central

    Teplitski, Max; Chen, Hancai; Rajamani, Sathish; Gao, Mengsheng; Merighi, Massimo; Sayre, Richard T.; Robinson, Jayne B.; Rolfe, Barry G.; Bauer, Wolfgang D.

    2004-01-01

    The unicellular soil-freshwater alga Chlamydomonas reinhardtii was found to secrete substances that mimic the activity of the N-acyl-l-homoserine lactone (AHL) signal molecules used by many bacteria for quorum sensing regulation of gene expression. More than a dozen chemically separable but unidentified substances capable of specifically stimulating the LasR or CepR but not the LuxR, AhyR, or CviR AHL bacterial quorum sensing reporter strains were detected in ethyl acetate extracts of C. reinhardtii culture filtrates. Colonies of C. reinhardtii and Chlorella spp. stimulated quorum sensing-dependent luminescence in Vibrio harveyi, indicating that these algae may produce compounds that affect the AI-2 furanosyl borate diester-mediated quorum sensing system of Vibrio spp. Treatment of the soil bacterium Sinorhizobium meliloti with a partially purified LasR mimic from C. reinhardtii affected the accumulation of 16 of the 25 proteins that were altered in response to the bacterium's own AHL signals, providing evidence that the algal mimic affected quorum sensing-regulated functions in this wild-type bacterium. Peptide mass fingerprinting identified 32 proteins affected by the bacterium's AHLs or the purified algal mimic, including GroEL chaperonins, the nitrogen regulatory protein PII, and a GTP-binding protein. The algal mimic was able to cancel the stimulatory effects of bacterial AHLs on the accumulation of seven of these proteins,