Estimating and correcting interference fringes in infrared spectra in infrared hyperspectral imaging

Short-term acclimation response of individual cells of Thalassiosira weissflogii was monitored by Synchrotron FTIR imaging over the span of 75 minutes. The cells, collected from batch cultures, were maintained in a constant flow of medium, at an irradiance of 120 μmol m−2 s−1 and at 20 °C. Multiple internal reflections due to the micro fluidic channel were modeled, and showed that fringes are additive sinusoids to the pure absorption of the other components of the system. Preprocessing of the hyperspectral cube (x, y, Abs(λ)) included removing spectral fringe using an EMSC approach. Principal component analysis of the time series of hyperspectral cubes showed macromolecular pool variations (carbohydrates, lipids and DNA/RNA) of less than 2% after fringe correction

Anaplerosis in microalgae

We describe the anapldrotic metabolism of alga

Toward Enhanced Fixation of CO2 in Aquatic Biomass: Focus on Microalgae

The need to reduce the CO2 footprint of human activities calls for the utilization of new means of production and new sources of products. Microalgae are a very promising source of a large variety of products, from fuels to chemicals for multiple industrial applications (e.g., dyes, pharmaceutical products, cosmetics, food and feed, new materials for high tech manufacture), and for processes such as wastewater treatment. Algae, as photosynthetic organisms, use light to energize the synthesis of organic matter and differently from most terrestrial plants, can be cultured on land that is not used for crop production. We describe the main factors contributing to microalgae productivity in artificial cultivation systems and discuss the research areas that still need investigation in order to pave the way to the generation of photosynthetic cell factories. We shall comment on the main caveats of the possible mode of improving photosynthetic efficiency and to optimize the partitioning of fixed C to products of commercial relevance. We address the problem of the selection of the appropriate strain and of the consequences of their diverse physiology and culture conditions for a successful commercial application. Finally, we shall provide state of the art information on cell factories chassis by means of synthetic biology approaches to produce chemicals of interest

Si decline and diatom evolution: Insights from physiological experiments

In today's oceans, diatoms are abundant and diverse primary producers distinguished by their silica shells. Although molecular clocks suggest that diatoms arose as much as 250 million years ago (Ma), the earliest known diatom fossils date from 190 Ma, leading to the suggestion that early diatoms were at best lightly silicified. By the Cretaceous Period, large circular (in cross section) diatoms with highly silicified frustules thrived in surface oceans, only later to be joined by species with elongated and thinner frustules, as well as lower SiO2 content. Decreased Si availability in surface oceans has been proposed as a principal driver of diatom evolution. Here, we investigate this through physiological experiments assessing the functional acclimation response of diatoms to reconstructed paleo-seawater. Four diatom species, differing in size and shape, were acclimated to reconstructed paleoenvironments mimicking Mesozoic/Cenozoic concentrations of nutrients in the presence of different Si regimes. When exposed to 500 mu M Si, all populations, save for that of Conticribra weissflogii, became more highly silicified; the higher Si content per cell at 500 mu M Si coincided with slower growth in small-sized cells. All species except C. weissflogii also showed lower photosynthetic efficiency as well as greater cell volume in comparison with diatoms acclimated to 205 or 25 mu M Si. Average cell stoichiometry correlates with cell shape, but not size; pennates, in particular Phaeodactylum tricornutum, showed an acclimatory response to Si regimes, modulating Si use efficiency (the lower the external Si concentrations, the higher the C and N quotas per Si).Experimental data suggest that in the densely silicified and bigger C. weissflogii grown at higher Si, diffusion of silicic acid across membranes made a larger contribution to Si uptake, saving energy which could be reallocated into growth. In contrast, for less highly silicified and smaller species, high energy costs of Si homeostasis needed to prevent the overaccumulation of intracellular Si limited growth. While our experimental species reacted individualistically to changing silica availability, with distinct levels of plasticity, selective pressure associated with the temporal decline in Si availability may well have favored elongated shapes. Modern, less silicified species are unable to exploit high Si concentrations

PHYSIOLOGICAL RESPONSES OF PHAEODACTYLUM TRICORNUTUM AND CYLINDROTHECA CLOSTERIUM (BACILLARIOPHYCEAE) TO DIFFERENT N-REGIMES

Poster PIV04 pg. 5

Role of phosphoenolpyruvate carboxylase in anaplerosis in the green microalga Dunaliella salina cultured under different nitrogen regimes

Anaplerosis plays a very important role in providing C for N assimilation. In green algae and higher plants, phosphoenolpyruvate carboxylase (PEPC, EC 4.1.1.31) is the main anaplerotic carboxylase. On this basis we hypothesize that N availability affects PEPC expression. In order to test this hypothesis, the model organism Dunaliella salina was cultured under a variety of N growth regimes. Our results show that the level of PEC activity was unaffected by the N form in which N was supplied to the cells, when N concentration was low (0.5–0.01 mM). When cells were adapted to growth at 5 mM N, however, PEPC activity on a per cell basis was substantially higher in NH4+-adapted cells as compared to their NO3–-adapted counterparts; however, the same difference was not observed on a protein basis. This notwithstanding, even at low N, PEPC of cells cultured in the presence of either NH4+ or NO3– appeared to differ in their molecular masses. These results suggest that cells adapted to different N-form express distinct PEPC isoforms. In addition to this, we observed that, in algae adapted to high (5 mM) NH4+ concentration, a PEPC isoform was induced that differed from the isoforms observed in algae adapted to lower concentrations of the same N-source. These findings lead us to conclude that the expression of PEPC isoforms in D. salina responds to the variation in the C-skeleton demand deriving from changes in the chemical form and availability of N

Taxonomy and growth conditions concur to determine the energetic suitability of fatty acid complements in algae

We hypothesize that, in algae, carbon allocation to lipids depends on the combined effects of the metabolic constraints imposed by the genotype (i.e., species-specific differences in composition) and on the acclimation responses (phenotype) to changes in the stoichiometry of available inorganic carbon (for photosynthesis) and nitrogen (primarily for amino acids, protein, and nucleic acid synthesis). We thus cultured three taxonomically distinct algae, the green alga Dunaliella salina, the diatom Thalassiosira pseudonana, and the dinoflagellate Protoceratium reticulatum, at four nitrate concentrations and constant inorganic carbon. Since energy availability also directly impacts carbon partitioning, we studied the effect of irradiance on the oil quality of P. reticulatum. We used Fourier transform infrared (FTIR) spectroscopy to study carbon allocation and biomass reduction level and gas chromatography for fatty acid analysis. The fatty acid complements of the three species were different; within each species, growth conditions substantially altered oil quality. We ranked the oils in terms of their suitability as biodiesels, using international standards as reference. We believe that this approach may help to identify the appropriate combination of taxa and culture conditions for algal biodiesel production and in general offers insight on carbon allocation to fatty acids