Kjersti Andresen

An Integrated Analysis of Molecular Acclimation to High Light in the Marine Diatom Phaeodactylum tricornutum

Photosynthetic diatoms are exposed to rapid and unpredictable changes in irradiance and spectral quality, and must be able to acclimate their light harvesting systems to varying light conditions. Molecular mechanisms behind light acclimation in diatoms are largely unknown. We set out to investigate the mechanisms of high light acclimation in Phaeodactylum tricornutum using an integrated approach involving global transcriptional profiling, metabolite profiling and variable fluorescence technique. Algae cultures were acclimated to low light (LL), after which the cultures were transferred to high light (HL). Molecular, metabolic and physiological responses were studied at time points 0.5 h, 3 h, 6 h, 12 h, 24 h and 48 h after transfer to HL conditions. The integrated results indicate that the acclimation mechanisms in diatoms can be divided into an initial response phase (0–0.5 h), an intermediate acclimation phase (3–12 h) and a late acclimation phase (12–48 h). The initial phase is recognized by strong and rapid regulation of genes encoding proteins involved in photosynthesis, pigment metabolism and reactive oxygen species (ROS) scavenging systems. A significant increase in light protecting metabolites occur together with the induction of transcriptional processes involved in protection of cellular structures at this early phase. During the following phases, the metabolite profiling display a pronounced decrease in light harvesting pigments, whereas the variable fluorescence measurements show that the photosynthetic capacity increases strongly during the late acclimation phase. We show that P. tricornutum is capable of swift and efficient execution of photoprotective mechanisms, followed by changes in the composition of the photosynthetic machinery that enable the diatoms to utilize the excess energy available in HL. Central molecular players in light protection and acclimation to high irradiance have been identified.

Modeling of light-dependent algal photosynthesis and growth: experiments with the Barents sea diatoms Thalassiosira nordenskioldii and Chaetoceros furcellatus

Abstract The models by Sakshaug et al (1989, Limnology and Oceanography, 34, 198–205) and Webb et al. (1974, Oecologia, 17, 281–291), for prediction of the gross growth rate of phytoplankton and short-term photosynthesis, respectively, have been modified on the basis of experiments with cultures of the centric diatoms Thalassiosira nordenskioeldii and Chaetoceros furcellatus grown at 0.5°C at combinations of two irradiances (25 and 400μmol m−2s−1) and two day-lengths (12 and 24 h). The models have one spectrum, °σ, which represents chlorophyll a (Chla) specific absorption of photosynthetically usable light, and introduces a factor q which represents Chla per PSU, functionally defined. The models describe phytoplankton growth in terms of physiologically relevant coefficients. A properly scaled fluorescence excitation spectrum (°F) represents a more appropriate estimate for °σ than the Chla-specific absorption spectrum °ac judging from calculations of Φmax (=αB/°σ). On the basis of °F, Φmax is 0.04 g-at C(mol photons)−1 for gross growth and about 0.05–0.08 for short-term carbon uptake (unfiltered samples). Calculations based on °ac yield values for Φmax which on average are 44% lower. P vs I (photosynthesis vs irradiance) parameters are relatively independent of day-length and highly dependent on growth irradiance. The product of q [mg Chla (mol PSU)−1] and τ (the minimum turnover time of the photosynthetic unit, h) increases 2–3-fold from high to low irradiance, thus PmB (=Φmax/qτ) and Ik(=1/qτ°σ)decreased. °F decreases from high to low irradiance. Carbon-specific dark respiration rates are Pigment ratios vary inversely with irradiance and day-length. The Chla:C ratio is particularly low under high, strong continuous light; Chlc: Chaa ratios are higher for shalde- than for light-adapted cells, while the converse is true for the ratio of the sum of the photoprotective pigments diadinoxanthin and diatoxanthin to Chla. The fucoxanthin: Chla ratio is virtually independent of the light regime. The two species are similar with respect to variations in growth rate (0.09–0.33 day−1 and Ik (31–36 vs 49–100 μmol m−2 s−1 at low and high irradiance, respectively). PmB and aB for growth as well as °F are systematically higher for C. furcellatus than for T. nordenskioeldii, while the product qτ is lower. C. furcellatus is considerably more plastic than T. nordenskioeldii with respect to pigment composition.

Molecular and Photosynthetic Responses to Prolonged Darkness and Subsequent Acclimation to Re-Illumination in the Diatom Phaeodactylum tricornutum

Photosynthetic diatoms that live suspended throughout the water column will constantly be swept up and down by vertical mixing. When returned to the photic zone after experiencing longer periods in darkness, mechanisms exist that enable the diatoms both to survive sudden light exposure and immediately utilize the available energy in photosynthesis and growth. We have investigated both the response to prolonged darkness and the re-acclimation to moderate intensity white irradiance (E = 100 µmol m−2 s−1) in the diatom Phaeodactylum tricornutum, using an integrated approach involving global transcriptional profiling, pigment analyses, imaging and photo-physiological measurements. The responses were studied during continuous white light, after 48 h of dark treatment and after 0.5 h, 6 h, and 24 h of re-exposure to the initial irradiance. The analyses resulted in several intriguing findings. Dark treatment of the cells led to 1) significantly decreased nuclear transcriptional activity, 2) distinct intracellular changes, 3) fixed ratios of the light-harvesting pigments despite a decrease in the total cell pigment pool, and 4) only a minor drop in photosynthetic efficiency (ΦPSII_max). Re-introduction of the cells to the initial light conditions revealed 5) distinct expression profiles for nuclear genes involved in photosynthesis and those involved in photoprotection, 6) rapid rise in photosynthetic parameters (α and rETRmax) within 0.5 h of re-exposure to light despite a very modest de novo synthesis of photosynthetic compounds, and 7) increasingly efficient resonance energy transfer from fucoxanthin chlorophyll a/c-binding protein complexes to photosystem II reaction centers during the first 0.5 h, supporting the observations stated in 6). In summary, the results show that despite extensive transcriptional, metabolic and intracellular changes, the ability of cells to perform photosynthesis was kept intact during the length of the experiment. We conclude that P. tricornutum maintains a functional photosynthetic apparatus during dark periods that enables prompt recovery upon re-illumination.

System responses to equal doses of photosynthetically usable radiation of blue, green, and red light in the marine diatom Phaeodactylum tricornutum.

Due to the selective attenuation of solar light and the absorption properties of seawater and seawater constituents, free-floating photosynthetic organisms have to cope with rapid and unpredictable changes in both intensity and spectral quality. We have studied the transcriptional, metabolic and photo-physiological responses to light of different spectral quality in the marine diatom Phaeodactylum tricornutum through time-series studies of cultures exposed to equal doses of photosynthetically usable radiation of blue, green and red light. The experiments showed that short-term differences in gene expression and profiles are mainly light quality-dependent. Transcription of photosynthesis-associated nuclear genes was activated mainly through a light quality-independent mechanism likely to rely on chloroplast-to-nucleus signaling. In contrast, genes encoding proteins important for photoprotection and PSII repair were highly dependent on a blue light receptor-mediated signal. Changes in energy transfer efficiency by light-harvesting pigments were spectrally dependent; furthermore, a declining trend in photosynthetic efficiency was observed in red light. The combined results suggest that diatoms possess a light quality-dependent ability to activate photoprotection and efficient repair of photodamaged PSII. In spite of approximately equal numbers of PSII-absorbed quanta in blue, green and red light, the spectral quality of light is important for diatom responses to ambient light conditions.