Elucidating biological processes has relied on the establishment of model organisms, many of which offer advantageous features such as rapid axenic growth, extensive knowledge of their physiological features and gene content, and the ease with which they can be genetically manipulated. The unicellular green alga Chlamydomonas reinhardtii has been an exemplary model that has enabled many scientific breakthroughs over the decades, especially in the fields of photosynthesis, cilia function and biogenesis, and the acclimation of photosynthetic organisms to their environment. Here, we discuss recent molecular/technological advances that have been applied to C. reinhardtii and how they have further fostered its development as a "flagship" algal system. We also explore the future promise of this alga in leveraging advances in the fields of genomics, proteomics, imaging, and synthetic biology for addressing critical future biological issues.

https://onlinelibrary.wiley.com/doi/pdfdirect/10.1111/jpy.13356

Chlamydomonas: Fast tracking from genomics.

Calcium (Ca2+)-dependent signalling plays a well-characterized role in the perception and response mechanisms to environmental stimuli in plant cells. In the context of a constantly changing environment, it is fundamental to understand how crop yield and microalgal biomass productivity are affected by external factors. Ca2+ signalling is known to be important in different physiological processes in microalgae but many of these signal transduction pathways still need to be characterized. Here, the role of compartment-specific Ca2+ signalling was investigated in Chlamydomonas reinhardtii in response to environmental stressors such as nutrient availability, osmotic stress, temperature fluctuations and carbon sensing. An in vivo single-cell imaging approach was adopted to directly visualize signalling processes at the level of specific subcellular compartments, using Chlamydomonas reinhardtii lines expressing a genetically encoded ratiometric Ca2+ indicator. Hyper-osmotic shock caused cytosolic and chloroplast Ca2+ elevations, whereas high temperature and inorganic carbon availability primarily induced Ca2+ transients in the chloroplast. In contrast, hypo-osmotic stress only induced Ca2+ elevations in the cytosol. The results herein reported show that compartment-specific signalling pathways are likely to play an important role in the response of Chlamydomonas to these stimuli providing new understanding of the mechanisms exploited by microalgae to respond to specific natural conditions. 

pdf/abiotic-stress-induced-chloroplast-and-cytosolic-ca2-47gig8smmx.pdf

Abiotic stress-induced chloroplast and cytosolic Ca2+ dynamics in the green alga Chlamydomonas reinhardtii

High temperatures impair plant/algal growth and reduce food/biofuel production, but the underlying mechanisms remain elusive. The unicellular green alga Chlamydomonas reinhardtii is a superior model to study heat responses in photosynthetic cells due to its fast growth rate, many similarities in cellular processes to land plants, simple and sequenced genome, and ample genetic/genomics resources. Chlamydomonas grows in light by photosynthesis and/or with supplied carbon source, acetate. The majority published research about Chlamydomonas heat responses used acetate-containing medium. How organic carbon sources affect heat responses is important for algal biofuel/bioproduct industry but understudied. We cultivated Chlamydomonas wild-type cultures under highly controlled conditions in photobioreactors at control of 25°C, moderate high temperature of 35°C, or acute high temperature of 40°C with and without constant acetate supply for 1- or 4-days. Our results showed that 35°C increased algal growth with constant acetate supply but reduced algal biomass and growth without sufficient acetate. The overlooked and dynamic effects of 35°C can be explained by induced carbon metabolisms, including acetate uptake/assimilation, glyoxylate cycle, gluconeogenesis pathways, and glycolysis. Acute high temperature at 40°C for more than 2 days was lethal to algal cultures with and without constant acetate supply. Our research provides insights to understand algal heat responses and help improve thermotolerance in photosynthetic cells. Highlight We revealed the overlooked, dynamic effects of moderate high temperature in algae depending on carbon availabilities and demonstrated the importance of carbon metabolism in thermotolerance of photosynthetic cells.

/pdf/moderate-high-temperature-is-beneficial-or-detrimental-k722qf10.pdf

Moderate High Temperature is Beneficial or Detrimental Depending on Carbon Availability in the Green Alga Chlamydomonas reinhardtii

Different high temperatures adversely affect crop and algal yields with various responses in photosynthetic cells. The list of genes required for thermotolerance remains elusive. Additionally, it is unclear how carbon source availability affects heat responses in plants and algae. We utilized the insertional, indexed, genome-saturating mutant library of the unicellular, eukaryotic green alga Chlamydomonas reinhardtii to perform genome-wide, quantitative, pooled screens under moderate (35°C) or acute (40°C) high temperatures with or without organic carbon sources. We identified heat-sensitive mutants based on quantitative growth rates and identified putative heat tolerance genes (HTGs). By triangulating HTGs with heat-induced transcripts or proteins in wildtype cultures and MapMan functional annotations, we present a high/medium-confidence list of 933 Chlamydomonas genes with putative roles in heat tolerance. Triangulated HTGs include those with known thermotolerance roles and novel genes with little or no functional annotation. About 50% of these high-confidence HTGs in Chlamydomonas have orthologs in green lineage organisms, including crop species. Arabidopsis thaliana mutants deficient in the ortholog of a high-confidence Chlamydomonas HTG were also heat sensitive. This work expands our knowledge of heat responses in photosynthetic cells and provides engineering targets to improve thermotolerance in algae and crops.

/pdf/high-throughput-identification-of-novel-heat-tolerance-genes-3tqg0yfe.pdf

High-throughput Identification of Novel Heat Tolerance Genes via Genome-wide Pooled Mutant Screens in the Model Green Alga Chlamydomonas reinhardtii

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High-throughput Identification of Novel Heat Tolerance Genes via Genome-wide Pooled Mutant Screens in the Model Green Alga Chlamydomonas reinhardtii

Moderate High Temperature is Beneficial or Detrimental Depending on Carbon Availability in the Green Alga Chlamydomonas reinhardtii