scholarly journals The Chloroplast of Chlamydomonas reinhardtii as a Testbed for Engineering Nitrogen Fixation into Plants

2021 ◽  
Vol 22 (16) ◽  
pp. 8806
Author(s):  
Marco Larrea-Álvarez ◽  
Saul Purton
Keyword(s):  
Chlamydomonas Reinhardtii ◽  
Metal Cluster ◽  
Essential Element ◽  
Crop Species ◽  
Photosynthetic Eukaryotes ◽  
Unicellular Green Alga ◽  
Coordinated Expression ◽  
Homocitrate Synthase ◽  
Eukaryotic Organisms ◽  
And Function

Eukaryotic organisms such as plants are unable to utilise nitrogen gas (N2) directly as a source of this essential element and are dependent either on its biological conversion to ammonium by diazotrophic prokaryotes, or its supply as chemically synthesised nitrate fertiliser. The idea of genetically engineering crops with the capacity to fix N2 by introduction of the bacterial nitrogenase enzyme has long been discussed. However, the expression of an active nitrogenase must overcome several major challenges: the coordinated expression of multiple genes to assemble an enzyme complex containing several different metal cluster co-factors; the supply of sufficient ATP and reductant to the enzyme; the enzyme’s sensitivity to oxygen; and the intracellular accumulation of ammonium. The chloroplast of plant cells represents an attractive location for nitrogenase expression, but engineering the organelle’s genome is not yet feasible in most crop species. However, the unicellular green alga Chlamydomonas reinhardtii represents a simple model for photosynthetic eukaryotes with a genetically tractable chloroplast. In this review, we discuss the main advantages, and limitations, of this microalga as a testbed for producing such a complex multi-subunit enzyme. Furthermore, we suggest that a minimal set of six transgenes are necessary for chloroplast-localised synthesis of an ‘Fe-only’ nitrogenase, and from this set we demonstrate the stable expression and accumulation of the homocitrate synthase, NifV, under aerobic conditions. Arguably, further studies in C. reinhardtii aimed at testing expression and function of the full gene set would provide the groundwork for a concerted future effort to create nitrogen-fixing crops.

scholarly journals Co-Expression Networks in the Green Alga Chlamydomonas reinhardtii Empower Gene Discovery and Functional Exploration

2020 ◽  
Author(s):  
Patrice A. Salomé ◽  
Sabeeha S. Merchant
Keyword(s):  
Chlamydomonas Reinhardtii ◽  
Green Alga ◽  
R Package ◽  
Sample Collection ◽  
Simple Method ◽  
Batch Cultures ◽  
Unicellular Green Alga ◽  
Function Discovery ◽  
Expression Potential ◽  
And Function

ABSTRACTThe unicellular green alga Chlamydomonas reinhardtii is a choice reference system for the study of photosynthesis, cilium assembly and function, lipid and starch metabolism and metal homeostasis. Despite decades of research, the functions of thousands of genes remain largely unknown, and new approaches are needed to categorically assign genes to cellular pathways. Growing collections of transcriptome and proteome data now allow a systematic approach based on integrative co-expression analysis. We used a dataset comprising 518 deep transcriptome samples derived from 58 independent experiments to identify potential co-expression relationships between genes. We visualized co-expression potential with the R package corrplot, to easily assess co-expression and anti-correlation between genes from manually-curated and community-generated gene lists. We extracted 400 high-confidence cilia-related genes at the intersection of multiple co-expressed lists, illustrating the power of our simple method. Surprisingly, Chlamydomonas experiments did not cluster according to an obvious pattern, suggesting an underappreciated variable during sample collection. One possible source of variation may stem from the strong clustering of nuclear genes as a function of their diurnal phase, even in samples collected in constant conditions, indicating substantial residual synchronization in batch cultures. We provide a step-by-step guide into the analysis of co-expression across Chlamydomonas transcriptome datasets to help foster gene function discovery.One-sentence summarywe reveal co-expression potential between Chlamydomonas genes and describe strong synchronization of cells grown in batch cultures as a possible source of underappreciated variation.

scholarly journals The Schizosaccharomyces pombe cho1+ Gene Encodes a Phospholipid Methyltransferase

Genetics ◽  
1998 ◽  
Vol 150 (2) ◽  
pp. 553-562
Author(s):  
Margaret I Kanipes ◽  
John E Hill ◽  
Susan A Henry
Keyword(s):  
Saccharomyces Cerevisiae ◽  
Sequence Analysis ◽  
Schizosaccharomyces Pombe ◽  
Gene Disruption ◽  
Structure And Function ◽  
Biosynthetic Enzyme ◽  
Gene Encoding ◽  
Complementation Groups ◽  
Eukaryotic Organisms ◽  
And Function

Abstract The isolation of mutants of Schizosaccharomyces pombe defective in the synthesis of phosphatidylcholine via the methylation of phosphatidylethanolamine is reported. These mutants are choline auxotrophs and fall into two unlinked complementation groups, cho1 and cho2. We also report the analysis of the cho1+ gene, the first structural gene encoding a phospholipid biosynthetic enzyme from S. pombe to be cloned and characterized. The cho1+ gene disruption mutant (cho1Δ) is viable if choline is supplied and resembles the cho1 mutants isolated after mutagenesis. Sequence analysis of the cho1+ gene indicates that it encodes a protein closely related to phospholipid methyltransferases from Saccharomyces cerevisiae and rat. Phospholipid methyltransferases encoded by a rat liver cDNA and the S. cerevisiae OPI3 gene are both able to complement the choline auxotrophy of the S. pombe cho1 mutants. These results suggest that both the structure and function of the phospholipid N-methyltransferases are broadly conserved among eukaryotic organisms.

scholarly journals Endogenous Fluctuations of DNA Topology in the Chloroplast of Chlamydomonas reinhardtii

1998 ◽  
Vol 18 (12) ◽  
pp. 7235-7242 ◽  
Author(s):  
Maria L. Salvador ◽  
Uwe Klein ◽  
Lawrence Bogorad
Keyword(s):  
Chlamydomonas Reinhardtii ◽  
Chloroplast Genome ◽  
Continuous Light ◽  
Dna Supercoiling ◽  
Dna Topology ◽  
Chloroplast Gene ◽  
Chloroplast Gene Expression ◽  
Endogenous Fluctuations ◽  
Unicellular Green Alga ◽  
In Cells

ABSTRACT DNA supercoiling in the chloroplast of the unicellular green algaChlamydomonas reinhardtii was found to change with a diurnal rhythm in cells growing in alternating 12-h dark–12-h light periods. Highest and lowest DNA superhelicities occurred at the beginning and towards the end of the 12-h light periods, respectively. The fluctuations in DNA supercoiling occurred concurrently and in the same direction in two separate parts of the chloroplast genome, one containing the genes psaB, rbcL, andatpA and the other containing the atpB gene. Fluctuations were not confined to transcribed DNA regions, indicating simultaneous changes in DNA conformation all over the chloroplast genome. Because the diurnal fluctuations persisted in cells kept in continuous light, DNA supercoiling is judged to be under endogenous control. The endogenous fluctuations in chloroplast DNA topology correlated tightly with the endogenous fluctuations of overall chloroplast gene transcription and with those of the pool sizes of most chloroplast transcripts analyzed. This result suggests that DNA superhelical changes have a role in the regulation of chloroplast gene expression in Chlamydomonas.

scholarly journals Photosymbiose — neuartige Anwendungen in der Biomedizin

BIOspektrum ◽  
2021 ◽  
Vol 27 (2) ◽  
pp. 202-204
Author(s):  
Myra N. Chávez ◽  
Benedikt Fuchs ◽  
Jörg Nickelsen
Keyword(s):  
Tissue Engineering ◽  
Chlamydomonas Reinhardtii ◽  
Green Alga ◽  
Recombinant Proteins ◽  
Biomedical Devices ◽  
Unicellular Green Alga ◽  
Surgical Sutures ◽  
Novel Strategy ◽  
Clinical Problems

AbstractWe have recently proposed a novel strategy named photosynthetic tissue engineering to overcome clinical problems due to hypoxia. The idea is based on transgenic photoautotrophic microorganisms that produce oxygen and at the same time secrete functional recombinant proteins into tissues. In particular, the unicellular green alga Chlamydomonas reinhardtii has successfully been used to boost the regenerative potential of several biomedical devices, such as dermal scaffolds and surgical sutures.

scholarly journals The structure and function of centriolar rootlets

2021 ◽  
Vol 134 (16) ◽  
Author(s):  
Robert Mahen
Keyword(s):  
Cellular Function ◽  
Structure And Function ◽  
Cellular Systems ◽  
Macromolecular Assemblies ◽  
Cellular Processes ◽  
Holistic Understanding ◽  
Eukaryotic Organisms ◽  
And Function ◽  
Knockout Animals

ABSTRACT To gain a holistic understanding of cellular function, we must understand not just the role of individual organelles, but also how multiple macromolecular assemblies function collectively. Centrioles produce fundamental cellular processes through their ability to organise cytoskeletal fibres. In addition to nucleating microtubules, centrioles form lesser-known polymers, termed rootlets. Rootlets were identified over a 100 years ago and have been documented morphologically since by electron microscopy in different eukaryotic organisms. Rootlet-knockout animals have been created in various systems, providing insight into their physiological functions. However, the precise structure and function of rootlets is still enigmatic. Here, I consider common themes of rootlet function and assembly across diverse cellular systems. I suggest that the capability of rootlets to form physical links from centrioles to other cellular structures is a general principle unifying their functions in diverse cells and serves as an example of how cellular function arises from collective organellar activity.

petD mRNA maturation in Chlamydomonas reinhardtii chloroplasts: role of 5' endonucleolytic processing

1994 ◽  
Vol 14 (9) ◽  
pp. 6180-6186
Author(s):  
W Sakamoto ◽  
N R Sturm ◽  
K L Kindle ◽  
D B Stern
Keyword(s):  
Chlamydomonas Reinhardtii ◽  
Fusion Gene ◽  
Deletion Mutants ◽  
Chloroplast Genes ◽  
Coding Region ◽  
Higher Plant ◽  
Glucuronidase Activity ◽  
Mrna Maturation ◽  
Processing Site ◽  
And Function

Complex processing of primary transcripts occurs during the expression of higher-plant chloroplast genes. In Chlamydomonas reinhardtii, most chloroplast genes appear to possess their own promoters, rather than being transcribed as part of multicistronic operons. By generating specific deletion mutants, we show that petD, which encodes subunit IV of the cytochrome b6/f complex, has an RNA processing site that is required for accumulation of monocistronic petD mRNA in petD promoter deletion mutants; in such mutants, transcription of petD originates from the upstream petA promoter. The 5' ends of transcripts initiated at the petD promoter are probably also generated by processing, since the 5' end of monocistronic petD mRNA is the same in wild-type strains as it is in the petD promoter mutants. The location and function of the processing site were further examined by inserting petD-uidA fusion genes into the chloroplast genome (uidA is an Escherichia coli gene that encodes beta-glucuronidase). When a promoterless petD-uidA fusion gene was inserted downstream of petA, a monocistronic uidA transcript accumulated, which was apparently initiated at the petA promoter and was processed at a site corresponding precisely to the petD mRNA 5' end. When a construct including only sequences downstream of +25 relative to the mature mRNA 5' end was inserted into the same site, a dicistronic petA-uidA transcript accumulated but no monocistronic uidA transcript could be detected, suggesting that a processing site lies at least partially within the region from -1 to +25. Beta-glucuronidase activity was not detected in transformants that accumulated only the dicistronic petA-uidA transcript, suggesting that the first 25 bp of the 5' untranslated region are required for translation initiation. One explanation for this translational defect is that Chlamydomonas chloroplasts cannot translate the second coding region of some dicistronic messages.

Chloroplast and Nuclear Mutations That Affect Rubisco Structure and Function in Chlamydomonas Reinhardtii

1992 ◽  
pp. 593-600 ◽  
Author(s):  
Robert J. Spreitzer ◽  
Graham Thow ◽  
Genhai Zhu ◽  
Zhixiang Chen ◽  
Cecilia Gotor ◽  
...  
Keyword(s):  
Chlamydomonas Reinhardtii ◽  
Structure And Function ◽  
Nuclear Mutations ◽  
And Function
Sign in / Sign up

Export Citation Format

Share Document