Changes in the photosynthetic apparatus and lipid droplet formation in Chlamydomonas reinhardtii under iron deficiency
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The unicellular photosynthetic alga Chlamydomonas reinhardtii was propagated in iron deficiency medium and patterns of growth, photosynthetic efficiency, lipid accumulation, as well as the expression of lipid biosynthetic and photosynthesis-related proteins were analysed and compared with iron-sufficient growth conditions. As expected, the photosynthetic rate was reduced (maximally after 4 days of growth) as a result of increased non-photochemical quenching (NPQ). Surprisingly, the stress-response protein LHCSR3 was expressed in conditions of iron deficiency that cause NPQ induction. In addition, the protein contents of both the PSI and PSII reaction centres were gradually reduced during growth in iron deficiency medium. Interestingly, the two generations of Fe deficiency cells could be able to recover the photosynthesis but the second generation cells recovered much slower as these cells were severely in shock. Analysis by flow cytometry with fluorescence-activated cell sorting and thin layer chromatography showed that iron deficiency also induced the accumulation of triacylglycerides (TAG), which resulted in the formation of lipid droplets. This was most significant between 48 and 72 h of growth. Dramatic increases in DGAT2A and PDAT1 levels were caused by iron starvation, which indicated that the biosynthesis of TAG had been increased. Analysis using gas chromatography mass spectrometry showed that levels of 16:0, 18:0, 18:2 and 18:3Δ9,12,15 fatty acids were significantly elevated. The results of this study highlight the genes/enzymes of Chlamydomonas that affect lipid synthesis through their influence on photosynthesis, and these represent potential targets of metabolic engineering to develop strains for biofuel production.
KeywordsElectron transport Iron deficiency LHCSR3 Major lipid droplet protein Photosystems Triacylglycerol
R.S was supported by the Council of Scientific and Industrial Research [No. 38(1279)/11/EMR-II and No. 38(1381)/14/EMR-II], Department of Biotechnology (BT/PR14964/BPA/118/137/2015) and DST-FIST, UGC-SAP, Govt. of India, for financial support. ED acknowledges the receipt of a UGC-RGNF fellowship. We thank Anthony H. C. Huang for the antibody against MLDP. We acknowledge Dr. John R. Gittins, University of Southampton, National Oceanography Center, Waterfront Road, Southampton, UK for his critical reading and also English correction.
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Conflict of interest
The authors declare that they have no conflict of interest.
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