Abstract

BackgroundUnder nitrogen deficiency situation, Nannochloropsis spp. accumulate large amounts of lipids in the form of triacylglycerides (TAG). Mechanisms of this process from the perspective of transcriptome and metabolome have been obtained previously, yet proteome analysis is still sparse which hinders the analysis of dynamic adaption to nitrogen deficiency. Here, proteomes for 3 h, 6 h, 12 h, 24 h, 48 h and 10th day of nitrogen deplete (N−) and replete (N+) conditions were obtained and integrated with previous transcriptome data for N. oceanica.ResultsPhysiological adaptations to N− not apparent from transcriptome data were unveiled: (a) abundance of proteins related to photosynthesis only slightly decreased in the first 48 h, indicating that photosynthesis is still working efficiently, and protein amounts adjust gradually with reduction in chloroplast size. (b) Most proteins related to the TCA cycle were strongly upregulated after 48 h under N−, suggesting that respiration is enhanced after 48 h and that TCA cycle efflux supports the carbon required for lipid synthesis. (c) Proteins related to lipid accumulation via the Kennedy pathway increased their abundance at 48 h, synchronous with the previously reported diversification of fatty acids after 48 h.ConclusionsThis study adds a proteome perspective on the major pathways for TAG accumulation in Nannochloropsis spp. Temporal changes of proteome exhibited distinct adaptation phases that are usually delayed relative to transcriptomic responses. Notably, proteome data revealed that photosynthesis and carbon fixation are still ongoing even after 48 h of N−. Moreover, sometimes completely opposite trends in proteome and transcriptome demonstrate the relevance of underexplored post-transcriptional regulation for N− adaptation.

Highlights

  • Under nitrogen deficiency situation, Nannochloropsis spp. accumulate large amounts of lipids in the form of triacylglycerides (TAG)

  • The temporal adaption to N− can be separated into three phases and eight functional clusters To characterize the physiology upon switch from normal culture condition to N− in N. oceanica IMET1, proteomes were compared among culture samples from 3 h, 6 h, 12 h, 24 h, 48 h and 10 days, which provided proteome profiles of N. oceanica IMET1 cells under differing nitrogen availability

  • By employing label-free relative quantification (LFQ) for the proteome data, a total of 4114 protein sequences were identified and quantified. This represents about 41% of all theoretically predicted 9915 proteins encoded in the genome of N. oceanica IMET1 [6, 9, 35, 36]

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Summary

Introduction

Nannochloropsis spp. accumulate large amounts of lipids in the form of triacylglycerides (TAG). Mechanisms of this process from the perspective of transcriptome and metabolome have been obtained previously, yet proteome analysis is still sparse which hinders the analysis of dynamic adaption to nitrogen deficiency. Under nitrogen deplete (N−) condition, Nannochloropsis spp. accumulate lipid in the form of triacylglycerides (TAG) that can make up to 60% of its dry weight [1, 2]. The diacylglycerol acyltransferases (DGAT) in the Kennedy pathway, responsible for the formation of TAG, are suspected pacemakers of TAG synthesis [4, 8]. A time-series transcriptomic and lipidomic dataset tracking IMET1 in N− revealed increased transcript abundance for seven putative DGAT isoforms [6]. Metabolic changes for protein, starch and TAG content monitored on the single-cell level in IMET1 were characterized with single-cell Raman spectra, which provide a powerful tool to screen strains or profile bioprocess [16]

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