scholarly journals The small RNA locus map for Chlamydomonas reinhardtii

PLoS ONE ◽  
2020 ◽  
Vol 15 (11) ◽  
pp. e0242516
Author(s):  
Sebastian Y. Müller ◽  
Nicholas E. Matthews ◽  
Adrian A. Valli ◽  
David C. Baulcombe
Keyword(s):  
Chlamydomonas Reinhardtii ◽  
Genome Stability ◽  
Higher Plants ◽  
Multiple Correspondence Analysis ◽  
Regulation Of Gene Expression ◽  
Transcriptional Gene Silencing ◽  
Evolutionary Divergence ◽  
Learning Approaches ◽  
Higher Plant ◽  
Systematic Classification

Small (s)RNAs play crucial roles in the regulation of gene expression and genome stability across eukaryotes where they direct epigenetic modifications, post-transcriptional gene silencing, and defense against both endogenous and exogenous viruses. It is known that Chlamydomonas reinhardtii, a well-studied unicellular green algae species, possesses sRNA-based mechanisms that are distinct from those of land plants. However, definition of sRNA loci and further systematic classification is not yet available for this or any other algae. Here, using data-driven machine learning approaches including Multiple Correspondence Analysis (MCA) and clustering, we have generated a comprehensively annotated and classified sRNA locus map for C. reinhardtii. This map shows some common characteristics with higher plants and animals, but it also reveals distinct features. These results are consistent with the idea that there was diversification in sRNA mechanisms after the evolutionary divergence of algae from higher plant lineages.

scholarly journals Mapping the Algal Secret Genome: The small RNA Locus Map for Chlamydomonas reinhardtii

2020 ◽  
Author(s):  
Sebastian Y. Müller ◽  
Nicholas E. Matthews ◽  
Adrian A. Valli ◽  
David C. Baulcombe
Keyword(s):  
Chlamydomonas Reinhardtii ◽  
Genome Stability ◽  
Higher Plants ◽  
Multiple Correspondence Analysis ◽  
Regulation Of Gene Expression ◽  
Transcriptional Gene Silencing ◽  
Evolutionary Divergence ◽  
Learning Approaches ◽  
Higher Plant ◽  
Systematic Classification

AbstractSmall (s)RNAs play crucial roles in the regulation of gene expression and genome stability across eukaryotes where they direct epigenetic modifications, post-transcriptional gene silencing, and defense against both endogenous and exogenous viruses. The green alga Chlamydomonas reinhardtii is a well-studied unicellular alga species with sRNA-based mechanisms that are distinct from those of land plants. It is, therefore, a good model to study sRNA evolution but a systematic classification of sRNA mechanisms is lacking in this and any other algae. Here, using data-driven machine learning approaches including Multiple Correspondence Analysis (MCA) and clustering, we have generated a comprehensively annotated and classified sRNA locus map for C. reinhardtii. This map shows some common characteristics with higher plants and animals, but it also reveals distinct features. These results are consistent with the idea that there was diversification in sRNA mechanisms after the evolutionary divergence of algae from higher plant lineages.

scholarly journals Condensation of Rubisco into a proto-pyrenoid in higher plant chloroplasts

2020 ◽  
Vol 11 (1) ◽  
Author(s):  
Nicky Atkinson ◽  
Yuwei Mao ◽  
Kher Xing Chan ◽  
Alistair J. McCormick
Keyword(s):  
Chlamydomonas Reinhardtii ◽  
Recent Work ◽  
Single Phase ◽  
Inorganic Carbon ◽  
Higher Plants ◽  
Initial Step ◽  
Operating Efficiency ◽  
Higher Plant ◽  
Spontaneous Condensation ◽  
Concentrating Mechanism

AbstractPhotosynthetic CO2 fixation in plants is limited by the inefficiency of the CO2-assimilating enzyme Rubisco. In most eukaryotic algae, Rubisco aggregates within a microcompartment known as the pyrenoid, in association with a CO2-concentrating mechanism that improves photosynthetic operating efficiency under conditions of low inorganic carbon. Recent work has shown that the pyrenoid matrix is a phase-separated, liquid-like condensate. In the alga Chlamydomonas reinhardtii, condensation is mediated by two components: Rubisco and the linker protein EPYC1 (Essential Pyrenoid Component 1). Here, we show that expression of mature EPYC1 and a plant-algal hybrid Rubisco leads to spontaneous condensation of Rubisco into a single phase-separated compartment in Arabidopsis chloroplasts, with liquid-like properties similar to a pyrenoid matrix. This work represents a significant initial step towards enhancing photosynthesis in higher plants by introducing an algal CO2-concentrating mechanism, which is predicted to significantly increase the efficiency of photosynthetic CO2 uptake.

scholarly journals Purification and characterization of high- and low-molecular-mass isoforms of phosphoenolpyruvate carboxylase from Chlamydomonas reinhardtii

1998 ◽  
Vol 331 (1) ◽  
pp. 201-209 ◽  
Author(s):  
Jean RIVOAL ◽  
William C. PLAXTON ◽  
David H. TURPIN
Keyword(s):  
Green Algae ◽  
Phosphoenolpyruvate Carboxylase ◽  
Higher Plants ◽  
Current Model ◽  
Catalytic Subunit ◽  
Dihydroxyacetone Phosphate ◽  
Evolutionary Divergence ◽  
Banana Fruit ◽  
Higher Plant ◽  
Green Algal

Phosphoenolpyruvate carboxylase (PEPC) is a key enzyme in the supply of carbon skeletons for the assimilation of nitrogen by green algae. Two PEPC isoforms with respective native molecular masses of 400 (PEPC1) and 650 (PEPC2) kDa have been purified from Chlamydomonas reinhardtiiCW-15 cc1883 (Chlorophyceae). SDS/PAGE, immunoblot and CNBr peptide-mapping analyses indicate the presence of the same 100 kDa PEPC catalytic subunit in both isoforms. PEPC1 is a homotetramer, whereas PEPC2 seems to be a complex between the PEPC catalytic subunit and other immunologically unrelated polypeptides of 50–70 kDa. Kinetic analyses indicate that these PEPC isoforms are (1) differentially regulated by pH, (2) activated by glutamine and dihydroxyacetone phosphate and (3) inhibited by glutamate, aspartate, 2-oxoglutarate and malate. These results are consistent with the current model for the regulation of anaplerotic carbon fixation in green algae, and demonstrate that green algal PEPCs are uniquely regulated by glutamine. Several techniques were used to assess the structural relationships between C. reinhardtiiPEPC and the higher plant or prokaryotic enzyme. Immunoblot studies using anti-(green algal or higher plant PEPC) IgGs suggested that green algal (C. reinhardtii, Selenastrum minutum), higher plant (maize, banana fruit, tobacco) and prokaryotic (Synechococcus leopoliensis, Escherichia coli)PEPCs have little or no immunological relatedness. Moreover, the N-terminal amino acid sequence of the C. reinhardtiiPEPC subunit did not have significant similarity to the highly conserved corresponding region in enzymes from higher plants, and CNBr cleavage patterns of green algal PEPCs were distinct from those of higher plant and cyanobacterial PEPCs. These results point to significant evolutionary divergence between green algal, higher plant and prokaryotic PEPCs.

scholarly journals Structure of a C2S2M2N2-type PSII–LHCII supercomplex from the green alga Chlamydomonas reinhardtii

2019 ◽  
Vol 116 (42) ◽  
pp. 21246-21255 ◽  
Author(s):  
Liangliang Shen ◽  
Zihui Huang ◽  
Shenghai Chang ◽  
Wenda Wang ◽  
Jingfen Wang ◽  
...  
Keyword(s):  
Energy Transfer ◽  
Chlamydomonas Reinhardtii ◽  
Green Alga ◽  
Higher Plants ◽  
Huge Number ◽  
Higher Plant ◽  
Low Light ◽  
Green Algal ◽  
Light Harvesting Antenna ◽  
Energy Transfer Pathway

Photosystem II (PSII) in the thylakoid membranes of plants, algae, and cyanobacteria catalyzes light-induced oxidation of water by which light energy is converted to chemical energy and molecular oxygen is produced. In higher plants and most eukaryotic algae, the PSII core is surrounded by variable numbers of light-harvesting antenna complex II (LHCII), forming a PSII–LHCII supercomplex. In order to harvest energy efficiently at low–light-intensity conditions under water, a complete PSII–LHCII supercomplex (C2S2M2N2) of the green alga Chlamydomonas reinhardtii (Cr) contains more antenna subunits and pigments than the dominant PSII–LHCII supercomplex (C2S2M2) of plants. The detailed structure and energy transfer pathway of the Cr-PSII–LHCII remain unknown. Here we report a cryoelectron microscopy structure of a complete, C2S2M2N2-type PSII–LHCII supercomplex from C. reinhardtii at 3.37-Å resolution. The results show that the Cr-C2S2M2N2 supercomplex is organized as a dimer, with 3 LHCII trimers, 1 CP26, and 1 CP29 peripheral antenna subunits surrounding each PSII core. The N-LHCII trimer partially occupies the position of CP24, which is present in the higher-plant PSII–LHCII but absent in the green alga. The M trimer is rotated relative to the corresponding M trimer in plant PSII–LHCII. In addition, some unique features were found in the green algal PSII core. The arrangement of a huge number of pigments allowed us to deduce possible energy transfer pathways from the peripheral antennae to the PSII core.

scholarly journals Melatonin as Master Regulator in Plant Growth, Development and Stress Alleviator for Sustainable Agricultural Production: Current Status and Future Perspectives

2020 ◽  
Vol 13 (1) ◽  
pp. 294
Author(s):  
Khadija Nawaz ◽  
Rimsha Chaudhary ◽  
Ayesha Sarwar ◽  
Bushra Ahmad ◽  
Asma Gul ◽  
...  
Keyword(s):  
Gene Expression ◽  
Plant Growth ◽  
Crop Production ◽  
Higher Plants ◽  
Regulation Of Gene Expression ◽  
Current Status ◽  
Master Regulator ◽  
Abiotic Stressors ◽  
Pathogenesis Related Protein ◽  
Growth Development

Melatonin, a multifunctional signaling molecule, is ubiquitously distributed in different parts of a plant and responsible for stimulating several physiochemical responses against adverse environmental conditions in various plant systems. Melatonin acts as an indoleamine neurotransmitter and is primarily considered as an antioxidant agent that can control reactive oxygen and nitrogen species in plants. Melatonin, being a signaling agent, induces several specific physiological responses in plants that might serve to enhance photosynthesis, growth, carbon fixation, rooting, seed germination and defense against several biotic and abiotic stressors. It also works as an important modulator of gene expression related to plant hormones such as in the metabolism of indole-3-acetic acid, cytokinin, ethylene, gibberellin and auxin carrier proteins. Additionally, the regulation of stress-specific genes and the activation of pathogenesis-related protein and antioxidant enzyme genes under stress conditions make it a more versatile molecule. Because of the diversity of action of melatonin, its role in plant growth, development, behavior and regulation of gene expression it is a plant’s master regulator. This review outlines the main functions of melatonin in the physiology, growth, development and regulation of higher plants. Its role as anti-stressor agent against various abiotic stressors, such as drought, salinity, temperatures, UV radiation and toxic chemicals, is also analyzed critically. Additionally, we have also identified many new aspects where melatonin may have possible roles in plants, for example, its function in improving the storage life and quality of fruits and vegetables, which can be useful in enhancing the environmentally friendly crop production and ensuring food safety.

A Simple Method for Removal of the Chlamydomonas reinhardtii Cell Wall Using a Commercially Available Subtilisin (Alcalase)

2018 ◽  
Vol 28 (4) ◽  
pp. 169-178 ◽  
Author(s):  
Hyun-Ju Hwang ◽  
Yong Tae Kim ◽  
Nam Seon Kang ◽  
Jong Won Han
Keyword(s):  
Cell Wall ◽  
Chlamydomonas Reinhardtii ◽  
Algal Cell ◽  
Chemical Properties ◽  
Transformation Method ◽  
Lytic Enzyme ◽  
Higher Plant ◽  
Time Saving ◽  
Simple Method ◽  
Cell Wall Disruption

The algal cell wall is a potent barrier for delivery of transgenes for genetic engineering. Conventional methods developed for higher plant systems are often unable to penetrate or remove algal cell walls owing to their unique physical and chemical properties. Therefore, we developed a simple transformation method for <i>Chlamydomonas reinhardtii</i> using commercially available enzymes. Out of 7 enzymes screened for cell wall disruption, a commercial form of subtilisin (Alcalase) was the most effective at a low concentration (0.3 Anson units/mL). The efficiency was comparable to that of gamete lytic enzyme, a protease commonly used for the genetic transformation of <i>C. reinhardtii</i>. The transformation efficiency of our noninvasive method was similar to that of previous methods using autolysin as a cell wall-degrading enzyme in conjunction with glass bead transformation. Subtilisin showed approximately 35% sequence identity with sporangin, a hatching enzyme of <i>C. reinhardtii</i>, and shared conserved active domains, which may explain the effective cell wall degradation. Our trans­formation method using commercial subtilisin is more reliable and time saving than the conventional method using autolysin released from gametes for cell wall lysis.

scholarly journals Selectable Markers and Reporter Genes for Engineering the Chloroplast of Chlamydomonas reinhardtii

Biology ◽  
2018 ◽  
Vol 7 (4) ◽  
pp. 46 ◽  
Author(s):  
Lola Esland ◽  
Marco Larrea-Alvarez ◽  
Saul Purton
Keyword(s):  
Chlamydomonas Reinhardtii ◽  
Higher Plants ◽  
Reporter Genes ◽  
Bioactive Metabolites ◽  
Cell Protein ◽  
Cell Factory ◽  
Expression Platform ◽  
Foreign Genes ◽  
Valuable Compounds ◽  
High Level

Chlamydomonas reinhardtii is a model alga of increasing interest as a cell factory for the production of valuable compounds, including therapeutic proteins and bioactive metabolites. Expression of foreign genes in the chloroplast is particularly advantageous as: (i) accumulation of product in this sub-cellular compartment minimises potential toxicity to the rest of the cell; (ii) genes can integrate at specific loci of the chloroplast genome (plastome) by homologous recombination; (iii) the high ploidy of the plastome and the high-level expression of chloroplast genes can be exploited to achieve levels of recombinant protein as high as 5% total cell protein; (iv) the lack of any gene silencing mechanisms in the chloroplast ensures stable expression of transgenes. However, the generation of C. reinhardtii chloroplast transformants requires efficient methods of selection, and ideally methods for subsequent marker removal. Additionally, the use of reporter genes is critical to achieving a comprehensive understanding of gene expression, thereby informing experimental design for recombinant applications. This review discusses currently available selection and reporter systems for chloroplast engineering in C. reinhardtii, as well as those used for chloroplast engineering in higher plants and other microalgae, and looks to the future in terms of possible new markers and reporters that will further advance the C. reinhardtii chloroplast as an expression platform.

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