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Diatoms are important components of marine ecosystems and contribute greatly to the world’s primary production. Despite their important roles in ecosystems, the molecular basis of how diatoms cope with oxidative stress caused by nutrient fluctuations remains largely unknown. Here, an isobaric tags for relative and absolute quantitation (iTRAQ) proteomic method was coupled with a series of physiological and biochemical techniques to explore oxidative stress- and cell fate decision-related cellular and metabolic responses of the diatom Thalassiosira pseudonana to nitrate (N) and inorganic phosphate (P) stresses. A total of 1151 proteins were detected; 122 and 56 were significantly differentially expressed from control under N- and P-limited conditions, respectively. In N-limited cells, responsive proteins were related to reactive oxygen species (ROS) accumulation, oxidative stress responses and cell death, corresponding to a significant decrease in photosynthetic efficiency, marked intracellular ROS accumulation, and caspase-mediated programmed cell death activation. None of these responses were identified in P-limited cells; however, a significant up-regulation of alkaline phosphatase proteins was observed, which could be the major contributor for P-limited cells to cope with ambient P deficiency. These findings demonstrate that fundamentally different metabolic responses and cellular regulations are employed by the diatom in response to different nutrient stresses and to keep the cells viable. [ABSTRACT FROM AUTHOR]

The extracellular polysaccharides (ECPS) released by diatoms have significant roles in marine ecosystems and have potential applications including drug-discovery and biopharmaceutical precursors. In this study, matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) technology was used in the structural analysis of the ECPS released by Thalassiosira pseudonana (Bacillariophyta). Three different deproteinization methods, the Sevag method, the trichloroacetic acid (TCA) method, and the enzymolysis method, were compared in the purification of ECPS. Our results suggested that TCA was the best deproteinization method among the three methods for subsequent MALDI-TOF MS investigation because of its high ECPS yield, protein removal ability and reliable MALDI-TOF MS fingerprint. The degree of polymerization (d.p.) profiles, the molecular weight of the ECPS and the distribution pattern of the polymers with different molecular mass were described from the MALDI-TOF MS spectra. This work represents the whole-level composition of the ECPS released by the diatom and has improved our knowledge of the structural characterization of ECPS. [ABSTRACT FROM AUTHOR]