Quantification of carbon flux distribution in the metabolic network of microalgae remains important to understand the complex interplay between energy metabolism, carbon fixation, and assimilation pathways. This is even more relevant with respect to cyclic metabolism of microalgae under light-dark cycle. In the present study, flux balance analysis (FBA) was carried out for an indigenous isolate Chlorella sp. FC2 IITG under photoautotrophic and heterotrophic growth conditions. A shift in intracellular flux distribution was predicted during transition from nutrient sufficient phase to nutrient starvation phase of growth. Further, dynamic flux analysis (dFBA) was carried out to capture light-dark metabolism over discretized pseudo steady state time intervals. Our key findings include the following: (i) unlike heterotrophic condition, oxidative pentose phosphate (PP) pathway, and Krebs cycle were relatively inactive under photoautotrophic growth; (ii) in both growth conditions, while transhydrogenation reaction was highly active, glyoxalate shunt was found to be nonoperative; (iii) flux distribution during transition period was marked with up regulation of carbon flux toward nongrowth associated (NGA) maintenance energy, oxidative phosphorylation, and photophosphorylation; (iv) redirection of carbon flux from polysaccharide and neutral lipid resulted in up regulation of Krebs cycle flux in the dark phase; (v) elevated glycolytic and acetyl-CoA flux were coupled with induction of neutral lipid during light cycle of the growth; (vi) significantly active photophosphorylation in the light phase was able to satisfy cellular energy requirement without need of oxidative PP pathway; and (vi) unlike static FBA, dFBA predicted an unaltered NGA maintenance energy of 1.5 mmol g(-1) DCW h(-1).

The present study reports evaluation of an indigenous microalgal isolate Chlorella sp. FC2 IITG as a potential candidate for biodiesel production. Characterization of the strain was performed under photoautotrophic, heterotrophic, and mixotrophic cultivation conditions. Further, an open-pond cultivation of the strain under outdoor conditions was demonstrated to evaluate growth performance and lipid productivity under fluctuating environmental parameters and in the presence of potential contaminants. The key findings were: (1) the difference in cultivation conditions resulted in significant variation in the biomass productivity (73-114 mg l⁻¹ day⁻¹) and total lipid productivity (35.02-50.42 mg l⁻¹ day⁻¹) of the strain; (2) nitrate and phosphate starvation were found to be the triggers for lipid accumulation in the cell mass; (3) open-pond cultivation of the strain under outdoor conditions resulted in biomass productivity of 44 mg l⁻¹ day⁻¹ and total lipid productivity of 10.7 mg l⁻¹ day⁻¹; (4) a maximum detectable bacterial contamination of 7 % of the total number of cells was recorded in an open-pond system; and (5) fatty acid profiling revealed abundance of palmitic acid (C16:0), oleic acid (C18:1) and linoleic acid (C18:2), which are considered to be the key elements for suitable quality biodiesel.

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Silicon (Si) is the most abundant element on earth after oxygen and is very important for plant growth under stress conditions. In the present study, we inspected the role of Si in the mitigation of the negative effect of salt stress at three concentrations (40 mM, 80 mM, and 120 mM NaCl) in two wheat varieties (KRL-210 and WH-1105) with or without Si (0 mM and 2 mM) treatment. Our results showed that photosynthetic pigments, chlorophyll stability index, relative water content, protein content, and carbohydrate content were reduced at all three salt stress concentrations in both wheat varieties. Moreover, lipid peroxidation, proline content, phenol content, and electrolyte leakage significantly increased under salinity stress. The antioxidant enzyme activities, like catalase and peroxidase, were significantly enhanced under salinity in both leaves and roots; however, SOD activity was drastically decreased under salt stress in both leaves and roots. These negative effects of salinity were more pronounced in WH-1105, as KRL-210 is a salt-tolerant wheat variety. On the other hand, supplementation of Si improved the photosynthetic pigments, relative water, protein, and carbohydrate contents in both varieties. In addition, proline content, MDA content, and electrolyte leakage were shown to decline following Si application under salt stress. It was found that applying Si enhanced the antioxidant enzyme activities under stress conditions. Si showed better results in WH-1105 than in KRL-210. Furthermore, Si was found to be more effective at a salt concentration of 120 mM compared to low salt concentrations (40 mM, 80 mM), indicating that it significantly improved plant growth under stressed conditions. Our experimental findings will open a new area of research in Si application for the identification and implication of novel genes involved in enhancing salinity tolerance.

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