A small volume bioassay to assess bacterial/phytoplankton co-culture using WATER-Pulse-Amplitude-Modulated (WATER-PAM) fluorometry

© 2015 Journal of Visualized Experiments. Conventional methods for experimental manipulation of microalgae have employed large volumes of culture (20 ml to 5 L), so that the culture can be subsampled throughout the experiment1–7. Subsampling of large volumes can be problematic for several reasons: 1) it causes variation in the total volume and the surface area:volume ratio of the culture during the experiment; 2) pseudo-replication (i.e., replicate samples from the same treatment flask8) is often employed rather than true replicates (i.e., sampling from replicate treatments); 3) the duration of the experiment is limited by the total volume; and 4) axenic cultures or the usual bacterial microbiota are difficult to maintain during long-term experiments as contamination commonly occurs during subsampling. The use of microtiter plates enables 1 ml culture volumes to be used for each replicate, with up to 48 separate treatments within a 12.65 x 8.5 x 2.2 cm plate, thereby decreasing the experimental volume and allowing for extensive replication without subsampling any treatment. Additionally, this technique can be modified to fit a variety of experimental formats including: bacterial-algal co-cultures, algal physiology tests, and toxin screening9–11. Individual wells with an alga, bacterium and/or co-cultures can be sampled for numerous laboratory procedures including, but not limited to: WATER-Pulse-Amplitude-Modulated (WATER-PAM) fluorometry, microscopy, bacterial colony forming unit (cfu) counts and flow cytometry. The combination of the microtiter plate format and WATER-PAM fluorometry allows for multiple rapid measurements of photochemical yield and other photochemical parameters with low variability between samples, high reproducibility and avoids the many pitfalls of subsampling a carboy or conical flask over the course of an experiment

The bacterial symbiont Phaeobacter inhibens Shapes the life history of its algal host emiliania huxleyi

© 2018 Bramucci, Labeeuw, Orata, Ryan, Malmstrom and Case. Marine microbes form host-associated biofilm communities that are shaped by complex interactions between bacteria and their host. The roseobacter Phaeobacter inhibens exploits both symbiotic and pathogenic niches while interacting with its microalgal host Emiliania huxleyi. During co-cultivation over extended periods with E. huxleyi, we show that P. inhibens selectively kills two host cell types, the diploid calcifying strain and the haploid flagellated strain. Meanwhile, various non-calcifying diploid strains are resistant to this pathogen or the pathogen is avirulent to this cell type. This differential pathogenesis has the potential of dramatically altering the composition of E. huxleyi blooms, which are typically dominated by the roseobacter-susceptible calcifying strain. This cell type makes calcite plates, which are an important sink in the marine carbon cycle and forms part of the marine paleobotanic record. P. inhibens kills the haploid cells, which have been proposed as critical to the survival of the algae, as they readily escape both eukaryotic predation and viral infection. Consequently, bacteria such as P. inhibens could influence E. huxleyi's life history by selective pathogenesis, thereby altering the composition of cell types within E. huxleyi populations and its bloom-bust lifestyle

Validation of a cationic polyacrylamide flocculant for the harvesting fresh and seawater microalgal biomass

© 2019 Elsevier B.V. A simple, efficient, and fast settling flocculation technique to harvest microalgal biomass was demonstrated using a proprietary cationic polyacrylamide flocculant for a freshwater (Chlorella vulgaris) and a marine (Phaeodactylum tricornutum) microalgal culture at their mid-stationary growth phase. The optimal flocculant doses were 18.9 and 13.7 mg/g of dry algal biomass for C. vulgaris and P. tricornutum, respectively (equivalent to 7 g per m3 of algal culture for both species). The obtained optimal dose was well corroborated with changes in cell surface charge, and culture solution optical density and turbidity. At the optimal dose, charge neutralization of 64 and 86% was observed for C. vulgaris and P. tricornutum algal cells, respectively. Algae recovery was independent of the culture solution pH in the range of pH 6 to 9. Algal biomass recovery was achieved of 100 and 90% for C vulgaris and P. tricornutum respectively, and over 98% medium recovery was achievable by simple decanting

Indole-3-acetic acid is produced by Emiliania huxleyi coccolith-bearing cells and triggers a physiological response in bald cells

© 2016 Labeeuw, Khey, Bramucci, Atwal, de la Mata, Harynuk and Case. Indole-3-acetic acid (IAA) is an auxin produced by terrestrial plants which influences development through a variety of cellular mechanisms, such as altering cell orientation, organ development, fertility, and cell elongation. IAA is also produced by bacterial pathogens and symbionts of plants and algae, allowing them to manipulate growth and development of their host. They do so by either producing excess exogenous IAA or hijacking the IAA biosynthesis pathway of their host. The endogenous production of IAA by algae remains contentious. Using Emiliania huxleyi, a globally abundant marine haptophyte, we investigated the presence and potential role of IAA in algae. Homologs of genes involved in several tryptophan-dependent IAA biosynthesis pathways were identified in E. huxleyi. This suggests that this haptophyte can synthesize IAA using various precursors derived from tryptophan. Addition of L-tryptophan to E. huxleyi stimulated IAA production, which could be detected using Salkowski's reagent and GC × GC-TOFMS in the C cell type (coccolith bearing), but not in the N cell type (bald). Various concentrations of IAA were exogenously added to these two cell types to identify a physiological response in E. huxleyi. The N cell type, which did not produce IAA, was more sensitive to it, showing an increased variation in cell size, membrane permeability, and a corresponding increase in the photosynthetic potential quantum yield of Photosystem II (PSII). A roseobacter (bacteria commonly associated with E. huxleyi) Ruegeria sp. R11, previously shown to produce IAA, was co-cultured with E. huxleyi C and N cells. IAA could not be detected from these co-cultures, and even when stimulated by addition of L-tryptophan, they produced less IAA than axenic C type culture similarly induced. This suggests that IAA plays a novel role signaling between different E. huxleyi cell types, rather than between a bacteria and its algal host

Microalgae-based carbon capture and utilization: A critical review on current system developments and biomass utilization

Carbon capture and utilization (CCU) is an emerging technology with commercial potential to convert atmospheric carbon dioxide (CO2) into net zero or negative emission products. In microalgae-based CCU, microalgae utilize CO2 and sunlight to generate biomass for commercial applications. This paper reviews the current state of microalgal culture development for CCU and highlights its potential contribution to addressing climate change challenges. Current microalgal culture systems have not been designed for high throughput biomass growth and carbon capture. Raceways, high-rate algal ponds, and photobioreactors are the most widely used for microalgal cultivation at a large-scale. The limitations of these systems are related to microalgal growth requirements. Ponds are operated at narrow depth to ensure sufficient light distribution and thus need a large land surface. CO2 gas needs to be in a dissolved form for efficient utilization by microalgae. Innovative system designs to achieve optimized distribution of light, nutrient, and CO2 utilization for enhanced biomass production are crucial to achieve large-scale CO2 capture by microalgae. Data corroborated in this review highlights several innovative techniques to deliver CO2 effectively and enhance light illumination to microalgal cells. Submerged and internal illuminations can enhance light distribution without compromising culture volume and land requirements. CO2 delivery technique selections mainly depend on CO2 sources. The carbonation column appears to be the best option regarding efficiency, easy operation, and simple design. The downstream processes of microalgal culture (i.e. harvesting, biomass utilization, and water reuse) are important to make microalgae-based CCU a significant contribution to global carbon mitigation solutions

Ancient origin of the biosynthesis of lignin precursors

BACKGROUND: Lignin plays an important role in plant structural support and water transport, and is considered one of the hallmarks of land plants. The recent discovery of lignin or its precursors in various algae has raised questions on the evolution of its biosynthetic pathway, which could be much more ancient than previously thought. To determine the taxonomic distribution of the lignin biosynthesis genes, we screened all publicly available genomes of algae and their closest non-photosynthetic relatives, as well as representative land plants. We also performed phylogenetic analysis of these genes to decipher the evolution and origin(s) of lignin biosynthesis. RESULTS: Enzymes involved in making p-coumaryl alcohol, the simplest lignin monomer, are found in a variety of photosynthetic eukaryotes, including diatoms, dinoflagellates, haptophytes, cryptophytes as well as green and red algae. Phylogenetic analysis of these enzymes suggests that they are ancient and spread to some secondarily photosynthetic lineages when they acquired red and/or green algal endosymbionts. In some cases, one or more of these enzymes was likely acquired through lateral gene transfer (LGT) from bacteria. CONCLUSIONS: Genes associated with p-coumaryl alcohol biosynthesis are likely to have evolved long before the transition of photosynthetic eukaryotes to land. The original function of this lignin precursor is therefore unlikely to have been related to water transport. We suggest that it participates in the biological defense of some unicellular and multicellular algae. REVIEWERS: This article was reviewed by Mark Ragan, Uri Gophna, Philippe Deschamps

Gestion des zones d'activité économique existantes. Première analyse de bonnes pratiques en matière de mixité des fonctions et de gestion parcimonieuse du sol

Rapport intermédiair

Global Trends of Usage of Chlorophyll Fluorescence and Projections for the Next Decade

Chlorophyll fluorescence is the most widely used set of techniques to probe photosynthesis and plant stress. Its great versatility has given rise to different routine methods to study plants and algae. The three main technical platforms are pulse amplitude modulation (PAM), fast rise of chlorophyll fluorescence, and fast repetition rate. Solar-induced fluorescence (SIF) has also gained interest in the last few years. Works have compared their advantages and their underlying theory, with many arguments advanced as to which method is the most accurate and useful. To date, no data has assessed the exact magnitude of popularity and influence for each methodology. In this work, we have taken the bibliometrics of the past decade for each of the four platforms, have evaluated the public scientific opinion toward each method, and possibly identified a geographical bias. We used various metrics to assess influence and popularity for the four routine platforms compared in this study and found that, overall, PAM currently has the highest values, although the more recent SIF has increased in popularity rapidly during the last decade. This indicates that PAM is currently one of the fundamental tools in chlorophyll fluorescence.</jats:p

Bioactive small molecules mediate microalgal-bacterial interactions

© Springer International Publishing AG 2017. Microalgae are a diverse group of photosynthetic microorganisms found throughout the eukaryote tree. Although unicellular, they have complex relationships with the bacteria that surround them. These interactions can range from obligate symbiosis, where the bacterium is required for host survival, to pathogenic, where the bacterial pathogen can kill the host alga. The nature of these algal-bacterial interactions appear to be tightly regulated by both algal and bacterial bioactive molecules, creating a complex system of chemical interactions through which these different species can chemically communicate with each other and directly alter the other physiology. In this way the bacterium is able to exploit (and manipulate) its host to become a more conducive habitat (e.g. algal phycosphere, aquatic biofilms, etc.) for bacterial survival. However, the identity of many of these small molecules and the mechanisms by which they control these exchanges are often overlooked or misunderstood. The ability to eavesdrop on the chemical cross talk occurring between algae and bacteria may open up a vast potential for new knowledge, relating to understanding bacterial-algal relationships, evolution and possibly hijacking this communication to better control microbes in commercial systems. This chapter outlines some of the known bioactive chemicals that mediate these microalgal-bacterial interactions, highlighting what is currently known about these systems and areas that need further investigation