Influence of Symbiont Species on the Glycerol and Glucose Pools in a Model Cnidarian-Dinoflagellate Symbiosis.

AbstractSymbiotic dinoflagellates in the family Symbiodiniaceae release mobile compounds (e.g., glucose, glycerol, amino acids, and lipids) to their host's tissues. Little is known about how different symbionts affect quantitative and qualitative differences in these compounds. We tested how symbiont identity affects glycerol and glucose pools in the tissues of the sea anemone Exaiptasia pallida ("Aiptasia"). We infected symbiont-free anemones with three different symbiotic dinoflagellates: Breviolum minutum isolated from our Aiptasia stock, B. minutum isolated from a different Aiptasia population, and the free-living Effrenium voratum. We measured free glycerol and glucose levels in host tissues under photosynthetic conditions, as well as metabolite release by these algae when freshly isolated from Aiptasia and incubated in a host tissue homogenate. Anemone tissues containing the stock B. minutum accumulated glycerol at a higher symbiont cell-specific rate than those containing the alternative B. minutum or E. voratum (e.g., at 9 h of light incubation: stock B. minutum, 4.05 × 10-5 nmol per algal cell; alternative B. minutum, 0.90 × 10-5 nmol per algal cell; E. voratum: 1.14 × 10-5 nmol per algal cell). All symbiotic hosts accumulated glucose between 1 and 12 h of light incubation. At 12 h, the symbiont cell-specific glucose content was 6-fold higher in anemone tissues that contained stock B. minutum than in those containing the alternative B. minutum (1.73 × 10-6 vs. 0.30 × 10-6 nmol per algal cell, respectively). All freshly isolated algae released glycerol and glucose when incubated in host homogenate except E. voratum, which did not release glycerol. These trends mirrored those in hospite. Our results suggest that, on an algal cell-specific basis, B. minutum isolated from the same Aiptasia stock provided better nutritional support to the host than did the two other algae, though this was not reflected in colonization success, highlighting the underlying complexity of host-symbiont recognition and symbiosis establishment in the cnidarian-dinoflagellate partnership.

Experiences with a researcher-centric ELN.

Electronic Laboratory Notebooks (ELNs) are progressively replacing traditional paper books in both commercial research establishments and academic institutions. University researchers require specific features from ELNs, given the need to promote cross-institutional collaborative working, to enable the sharing of procedures and results, and to facilitate publication. The LabTrove ELN, which we use as our exemplar, was designed to be researcher-centric (i.e., not only aimed at the individual researcher's basic needs rather than to a specific institutional or subject or disciplinary agenda, but also able to be tailored because it is open source). LabTrove is being used in a heterogeneous set of academic laboratories, for a range of purposes, including analytical chemistry, X-ray studies, drug discovery and a biomaterials project. Researchers use the ELN for recording experiments, preserving data collected, and for project coordination. This perspective article describes the experiences of those researchers from several viewpoints, demonstrating how a web-based open source electronic notebook can meet the diverse needs of academic researchers.

Formyl group modification of chlorophyll a: a major evolutionary mechanism in oxygenic photosynthesis.

We discuss recent advances in chlorophyll research in the context of chlorophyll evolution and conclude that some derivations of the formyl side chain arrangement of the porphyrin ring from that of the Chl a macrocycle can extend the photosynthetic active radiation (PAR) of these molecules, for example, Chl d and Chl f absorb light in the near-infrared region, up to ∼750 nm. Derivations such as this confer a selective advantage in particular niches and may, therefore, be beneficial for photosynthetic organisms thriving in light environments with particular light signatures, such as red- and near-far-red light-enriched niches. Modelling of formyl side chain substitutions of Chl a revealed yet unidentified but theoretically possible Chls with a distinct shift of light absorption properties when compared to Chl a.

Electrogenic plasma membrane H+-ATPase activity using voltage sensitive dyes.

Fast responding voltage sensitive dyes, RH421 and di-4-ASPBS, were used to study the electrogenic properties of plant plasma membrane proton pumps on sealed plasma membrane vesicles extracted by two-phase partitioning from Beta vulgaris and Avena sativa cv Swan root material. Fluorescence spectroscopy in the presence of the dye RH421 (10.8 nM) was sufficiently sensitive to detect electrogenic activity of the extracted plant vesicles. The dye detection system could detect inhibition of electrogenic activity of vesicles by vanadate (75 µM) and stimulation by nigericin (0.5 µM). The newly developed dye di-4-ASPBS was less sensitive to detecting the electrogenic proton pump activity. This study represents an important innovation in plant biophysics as this class of fast responding voltage sensitive dyes have never to our knowledge been used to study electrogenic proton pump activity derived from plant membranes and represents a novel approach for carrying out such studies.

Chromatic photoacclimation, photosynthetic electron transport and oxygen evolution in the chlorophyll d-containing oxyphotobacterium Acaryochloris marina.

Changes in photosynthetic pigment ratios showed that the Chlorophyll d-dominated oxyphotobacterium Acaryochloris marina was able to photoacclimate to different light regimes. Chl d per cell were higher in cultures grown under low irradiance and red or green light compared to those found when grown under high white light, but phycocyanin/Chl d and carotenoid/Chl d indices under the corresponding conditions were lower. Chl a, considered an accessory pigment in this organism, decreased respective to Chl d in low irradiance and low intensity non-white light sources. Blue diode PAM (Pulse Amplitude Modulation) fluorometry was able to be used to measure photosynthesis in Acaryochloris. Light response curves for Acaryochloris were created using both PAM and O(2) electrode. A linear relationship was found between electron transport rate (ETR), measured using a PAM fluorometer, and oxygen evolution (net and gross photosynthesis). Gross photosynthesis and ETR were directly proportional to one another. The optimum light for white light (quartz halogen) was about 206+/-51 micromol m(-2) s(-1) (PAR) (Photosynthetically Active Radiation), whereas for red light (red diodes) the optimum light was lower (109+/-27 micromol m(-2) s(-1) (PAR)). The maximum mean gross photosynthetic rate of Acaryochloris was 73+/-7 micromol mg Chl d(-1) h(-1). The gross photosynthesis/respiration ratio (P(g)/R) of Acaryochloris under optimum conditions was about 4.02+/-1.69. The implications of our findings will be discussed in relation to how photosynthesis is regulated in Acaryochloris.

The major light-harvesting pigment protein of Acaryochloris marina.

The major light-harvesting protein complex containing chlorophyll (Chl) d was isolated from Acaryochloris marina thylakoid membranes. Isolation was achieved by detergent solubilisation followed by separation on 6-40% sucrose gradients using ultracentrifugation. The best Chl d yield (70%) used 0.3% dodecyl maltoside, 0.15% octyl glucoside, 0.05% zwittergent 3-14 with the detergent:total Chl d ratio around 10:1 (w/w). Characterisation of the light-harvesting pigment protein complex (lhc) involved non-denaturing electrophoresis, SDS-PAGE, absorbance and fluorescence spectroscopy. The main polypeptide in the lhc was shown to be ca. 34 kDa and to contain Chl d and Chl a, indicating that the Acaryochloris lhc is similar to that of prochlorophytes. The Chl a level varied with the culture conditions, which is consistent with previous findings.