Photosynthesis Research

, Volume 139, Issue 1–3, pp 253–266 | Cite as

Changes in the photosynthetic apparatus and lipid droplet formation in Chlamydomonas reinhardtii under iron deficiency

  • Elsinraju Devadasu
  • Dinesh Kumar Chinthapalli
  • Nisha Chouhan
  • Sai Kiran Madireddi
  • Girish Kumar Rasineni
  • Prabhakar Sripadi
  • Rajagopal SubramanyamEmail author
Original Article


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.


Electron 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.

Compliance with ethical standards

Conflict of interest

The authors declare that they have no conflict of interest.

Supplementary material

11120_2018_580_MOESM1_ESM.docx (310 kb)
Supplementary material 1 (DOCX 310 KB)


  1. Andaluz S, Millan AF, De las Rivas J, Aro EM, Abadía J (2006) Proteomic profiles of thylakoid membranes and changes in response to iron deficiency. Photosynth Res 89:141–155CrossRefGoogle Scholar
  2. Aro EM, Virgin I, Andersson B (1993) Photoinhibition of photosystem II. Inactivation, protein damage and turnover. Biochim Biophys Acta 1143:113–134CrossRefGoogle Scholar
  3. BenMoussa-Dahmen I, Chtourou H, Rezgui F, Sayadi S, Dhouib A (2016) Salinity stress increases lipid, secondary metabolites and enzyme activity in Amphora subtropica and Dunaliella sp. for biodiesel production. Bioresour Technol 218:816–825CrossRefGoogle Scholar
  4. Bibby TS, Nield J, Barber J (2001) Iron deficiency induces the formation of an antenna ring around trimeric photosystem I in cyanobacteria. Nature 412:743–745CrossRefGoogle Scholar
  5. Boekema EJ, Hifney A, Yakushevska AE, Piotrowski M, Keegstra W, Berry S, Michel KP, Pistorius EK, Kruip J (2001) A giant chlorophyll-protein complex induced by iron deficiency in cyanobacteria. Nature 412:745–753CrossRefGoogle Scholar
  6. Bonente G, Ballottari M, Truong TB, Morosinotto T, Ahn TK, Fleming GR, Niyogi KK, Bassi R (2011) Analysis of LHCSR3, a protein essential for feedback de-excitation in the green alga Chlamydomonas reinhardtii. PLoS Biol 9:e1000577CrossRefGoogle Scholar
  7. Borowitzka MA, Moheimani NR (2013) Sustainable biofuels from algae. Mitig Adapt Strateg Global Change 18:13–25CrossRefGoogle Scholar
  8. Boyle NR, Page MD, Liu B, Blaby IK, Casero D, Kropat J, Cokus SJ, Hong-Hermesdorf A, Shaw J, Karpowicz SJ (2012) Three acyltransferases and nitrogen-responsive regulator are implicated in nitrogen starvation-induced triacylglycerol accumulation in Chlamydomonas. J Biol Chem 287:15811–15825CrossRefGoogle Scholar
  9. Busch A, Rimbauld B, Naumann B, Rensch S, Hippler M (2008) Ferritin is required for rapid remodeling of the photosynthetic apparatus and minimizes photo-oxidative stress in response to iron availability in Chlamydomonas reinhardtii. Plant J 55:201–211CrossRefGoogle Scholar
  10. Cakmak ZE, Olmez TT, Cakmak T, Menemen Y, Tekinay T (2014) Induction of triacylglycerol production in Chlamydomonas reinhardtii: comparative analysis of different element regimes. Biores Technol 155:379–387CrossRefGoogle Scholar
  11. Chauhan D, Folea IM, Jolley CC, Kouril R, Lubner CE, Lin S, Kolber D, Wolfe-Simon F, Golbeck JH, Boekema EJ, Fromme P (2011) A novel photosynthetic strategy for adaptation to low-iron aquatic environments. Biochemistry 50:686–692CrossRefGoogle Scholar
  12. Chen W, Zhang C, Song L, Sommerfeld M, Hu Q (2009) A high throughput Nile red method for quantitative measurement of neutral lipids in microalgae. J Microbiol Methods 77:41–47CrossRefGoogle Scholar
  13. Chen M, Tang H, Ma H, Holland TC, Ng KYS, Salley SO (2011) Effect of nutrients on growth and lipid accumulation in the green algae Dunaliella tertiolecta. Biores Technol 102:1649–1655CrossRefGoogle Scholar
  14. Chen L, Ding C, Zhao X, Xu J, Mohammad AA, Wang S, Ding Y (2015) Differential regulation of proteins in rice (Oryza sativa L.) under iron deficiency. Plant Cell Rep 34:83–96CrossRefGoogle Scholar
  15. Chisti Y (2007) Biodiesel from microalgae. Biotechnol Adv 25:294–306CrossRefGoogle Scholar
  16. Correa-Galvis V, Redekop P, Guan K, Griess A, Truong TB, Wakao S, Niyogi KK, Jahns P (2016) Photosystem II subunit PsbS is involved in the induction of LHCSR protein-dependent energy dissipation in Chlamydomonas reinhardtii. J Biol Chem 291:17478–17565CrossRefGoogle Scholar
  17. Cournac L, Latouche G, Cerovic Z, Redding K, Ravenel J, Peltier G (2002) In vivo interactions between photosynthesis, mitorespiration, and chlororespiration in Chlamydomonas reinhardtii. Plant Physiol 129:1921–1929CrossRefGoogle Scholar
  18. Devadasu ER, Madireddi SK, Nama S, Subramanyam R (2016) Iron deficiency cause changes in photochemistry, thylakoid organization, and accumulation of photosystem II proteins in Chlamydomonas reinhardtii. Photosynth Res 130:469–478CrossRefGoogle Scholar
  19. Fan J, Andre C, Xu C (2011) A chloroplast pathway for the de novo biosynthesis of triacylglycerol in Chlamydomonas reinhardtii. FEEB Lett 585:1985–1991CrossRefGoogle Scholar
  20. Glaesener AG, Merchant SS, Blaby-Haas CE (2013) Iron economy in Chlamydomonas reinhardtii. Front Plant Sci 4:337–349CrossRefGoogle Scholar
  21. Huang NL, Huang MD, Chen TL, Huang AH (2013) Oleosin of subcellular lipid droplets evolved in green algae. Plant Physiol 161:1862–1874CrossRefGoogle Scholar
  22. Ivanov AG, Krol M, Sveshnikov D, Selstam E, Sandström S, Koochek M, Park Y, Vasil’ev S, Bruce D, Öquist G, Huner NPA (2006) Iron deficiency in cyanobacteria causes monomerization of photosystem I trimers and reduces the capacity for state transitions and the effective absorption cross section of photosystem I in vivo. Plant Physiol 141:1436–1445CrossRefGoogle Scholar
  23. Ivanov AG, Krol M, Sveshnikov D, Selstam E, Sane PV, Sveshnikov D, Park Y, Öquist G, Huner NPA (2007) The induction of CP43′ by Iron-stress in Synechococcus sp. PCC 7942 associated with carotenoid accumulation and enhanced fatty acid unsaturation. Biochim Biophys Acta 1767:807–813CrossRefGoogle Scholar
  24. James Gabriel O, Hocart Charles H, Hillier W, Chen H, Kordbacheh F, Price D, Djordjevic G, Michael A (2011) Fatty acid profiling of Chlamydomonas reinhardtii under nitrogen deprivation. Biores Technol 102:3343–3351CrossRefGoogle Scholar
  25. Juergens MT, Deshpande RR, Lucker BF, Park JJ, Wang H, Gargouri M, Holguin FO, Disbrow B, Schaub T, Skepper JN, Kramer DM, Gang DR, Hicks LM, Shachar-Hill Y (2015) The regulation of photosynthetic structure and function during nitrogen deprivation in Chlamydomonas reinhardtii. Plant Physiol 167:558–631CrossRefGoogle Scholar
  26. Kobayashi N, Noel EA, Barnes A, Rosenberg J, DiRusso C, Black P, Oyler GA (2013) Rapid detection and quantification of triacylglycerol by HPLC-ELSD in Chlamydomonas reinhardtii and Chlorella strains. Lipids 48:1035–1049CrossRefGoogle Scholar
  27. Kodru S, Malavath T, Devadasu E, Nellaepalli S, Stirbet A, Subramanyam R, Govindjee (2015) The slow S to M rise of chlorophyll a fluorescence reflects transition from state 2 to state 1 in the green alga Chlamydomonas reinhardtii. Photosynth Res 125:219–231CrossRefGoogle Scholar
  28. Kosourov S, Patrusheva E, Ghirardi L, Seibert M, Tsygankov A (2007) A comparison of hydrogen photoproduction by sulfur-deprived Chlamydomonas reinhardtii under different growth conditions. J Biotechnol 128:776–787CrossRefGoogle Scholar
  29. Kropat J, Hong-Hermesdorf A, Casero D, Ent P, Castruita M, Pellegrini M, Merchant SS, Malasarn D (2011) A revised mineral nutrient supplement increases biomass and growth rate in Chlamydomonas reinhardtii. Plant J 66:770–780CrossRefGoogle Scholar
  30. Lewis T, Nichols PD, McMeekin TA (2000) Evaluation of extraction methods for recovery of fatty acids from lipid-producing microheterotrophs. J Microbiol Methods 43:107–116CrossRefGoogle Scholar
  31. Li X, Moellering ER, Liu B, Johnny C, Fedewa M, Sears BB, Kuo MH, Benning C (2012a) A galactoglycerolipid lipase is required for triacylglycerol accumulation and survival following nitrogen deprivation in Chlamydomonas reinhardtii. Plant Cell 24:4670–4686CrossRefGoogle Scholar
  32. Li X, Benning C, Kuo MH (2012b) Rapid triacylglycerol turnover in Chlamydomonas reinhardtii requires a lipase with broad substrate specificity. Eukaryot Cell 11:1451–1462CrossRefGoogle Scholar
  33. Merchant SS et al (2007) The Chlamydomonas genome reveals the evolution of key animal and plant functions. Science 318:245–250CrossRefGoogle Scholar
  34. Moody JW, McGinty CM, Quinn JC (2014) Global evaluation of biofuel potential from microalgae. Proc Natl Acad Sci USA 111:8691–8697CrossRefGoogle Scholar
  35. Moseley JL, Allinger T, Herzog S, Hoerth P, Wehinger E, Merchant S, Hippler M (2002) Adaptation to Fe-deficiency requires remodeling of the photosynthetic apparatus. EMBO J 21:6709–6720CrossRefGoogle Scholar
  36. Msilini N, Zaghdoudi M, Govindachary S, Lachaaˆl M, Ouerghi Z (2011) Inhibition of photosynthetic oxygen evolution and electron transfer from the quinone acceptor Q A to Q B by iron deficiency. Photosynth Res 107:247–256CrossRefGoogle Scholar
  37. Naumann B, Stauber EJ, Busch A, Sommer F, Hippler M (2005) N-terminal processing of Lhca3 Is a key step in remodeling of the photosystem I-light-harvesting complex under iron deficiency in Chlamydomonas reinhardtii. J Biol Chem 280:20431–20441CrossRefGoogle Scholar
  38. Naumann B, Busch A, Allmer J, Ostendorf E, Zeller M, Kirchhoff H, Hippler M (2007) Comparative quantitative proteomics to investigate the remodeling of bioenergetic pathways under iron deficiency in Chlamydomonas reinhardtii. Proteomics 7:3964–3979CrossRefGoogle Scholar
  39. Neelam S, Subramanyam R (2013) Alteration of photochemistry and protein degradation of photosystem II from Chlamydomonas reinhardtii under high salt grown cells. J Photochem Photobiol B 124:63–70CrossRefGoogle Scholar
  40. Nguyen HM, Baudet M, Cuine S, Adriano JM, Barthe D, Billon E, Bruley C, Beisson F, Peltier G, Ferro M, Li-Beisson Y (2011) Proteomic profiling of oil bodies isolated from the unicellular green microalga Chlamydomonas reinhardtii: with focus on proteins involved in lipid metabolism. Proteomics 11:4266–4273CrossRefGoogle Scholar
  41. Nguyen HM, Cuine S, Beyly-Adriano A, Legeret B, Billon E, Auroy P, Beisson F, Peltier G, Li-Beisson Y (2013) The green microalga Chlamydomonas reinhardtii has a single omega-3 fatty acid desaturase that localizes to the chloroplast and impacts both plastidic and extraplastidic membrane lipids. Plant Physiol 163:914–928CrossRefGoogle Scholar
  42. Peers G, Truong TB, Ostendorf E, Busch A, Elrad D, Grossman AR, Hippler M, Niyogi KK (2009) An ancient light-harvesting protein is critical for the regulation of algal photosynthesis. Nature 462:518–521CrossRefGoogle Scholar
  43. Petroutsos D, Terauchi AM, Busch A, Hirschmann I, Merchant SS, Finazzi G, Hippler M (2009) PGRL1 participates in iron-induced remodeling of the photosynthetic apparatus and in energy metabolism in Chlamydomonas reinhardtii. J Biol Chem 284:32770–32781CrossRefGoogle Scholar
  44. Porra RJ, Thompson WA, Kriedemann PE (1989) Determination of accurate extinction coefficients and simultaneous equations for assaying chlorophylls a and b extracted with four different solvents: verification of the concentration of chlorophyll standards by atomic absorption spectroscopy. Biochim Biophys Acta 975:384–394CrossRefGoogle Scholar
  45. Ravenel JA (1988) The iron and molybdenum use efficiencies of plant growth with different energy, carbon and nitrogen sources. New Phytol 109:279–287CrossRefGoogle Scholar
  46. Schansker G, Strasser RJ (2005) Quantification of non-Q B-reducing centers in leaves using a far-red pre-illumination. Photosynth Res 84:145–151CrossRefGoogle Scholar
  47. Stirbet A, Govindjee (2011) On the relation between the Kautsky effect (chlorophyll a fluorescence induction) and Photosystem II: basics and applications of the OJIP fluorescence transient. J Photochem Photobiol B 104:236–257CrossRefGoogle Scholar
  48. Stirbet A, Govindjee (2012) Chlorophyll a fluorescence induction: understanding the thermal phase, the J-I-P rise. Photosynth Res 113:15–61CrossRefGoogle Scholar
  49. Terauchi AM, Peers G, Kobayashi MC, Niyogi KK, Merchant SS (2010) Trophic status of Chlamydomonas reinhardtii influences the impact of iron deficiency on photosynthesis. Photosynth Res 105:39–49CrossRefGoogle Scholar
  50. Timperio AM, D’Amici GM, Barta C, Loreto F, Zolla L (2007) Proteomics, pigment composition, and organization of thylakoid membranes in iron-deficient spinach leaves. J Exp Bot 58:3695–3710CrossRefGoogle Scholar
  51. Towbin H, Staehelin T, Gordon J (1979) Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: procedure and some applications. Proc Natl Acad Sci USA 76:4350–4354CrossRefGoogle Scholar
  52. Urzica EI, Vieler A, Hong-Hermesdorf A, Page MD, Casero D, Gallaher SD, Kropat J, Pellegrini M, Benning C, Merchant SS (2013) Remodeling of membrane lipids in iron-starved Chlamydomonas. J Biol Chem 288:30246–30258CrossRefGoogle Scholar
  53. Velmurugan N, Sung M, Yim SS, Park MS, Yang JW, Jeong KJ (2014) Systematically programmed adaptive evolution reveals potential role of carbon and nitrogen pathways during lipid accumulation in Chlamydomonas reinhardtii. Biotechnol Biofuel 7:117–132Google Scholar
  54. Wellburn AR (1994) The spectral determination of chlorophylls a and b, as well as total carotenoids, using various solvents with spectrophotometers of different resolution. J Plant Physiol 144:307–313CrossRefGoogle Scholar
  55. Yadavalli V, Jolley CC, Malleda C, Thangaraj B, Fromme P, Subramanyam R (2012a) Alteration of proteins and pigments influence the function of photosystem I under iron deficiency from Chlamydomonas reinhardtii. PLoS ONE 7:e35084CrossRefGoogle Scholar
  56. Yadavalli V, Neelam S, Rao ASVC, Reddy AR, Subramanyam R (2012b) Differential degradation of photosystem I subunits under iron deficiency in rice. J Plant Physiol 169:753–759CrossRefGoogle Scholar
  57. Yoon K, Han D, Li Y, Sommerfeld M, Hu Q (2012) Phospholipid:diacylglycerol acyltransferase is a multifunctional enzyme involved in membrane lipid turnover and degradation while synthesizing triacylglycerol in the unicellular green microalga Chlamydomonas reinhardtii. Plant Cell 24:3708–3724CrossRefGoogle Scholar
  58. Zhang M, Fan J, Taylor DC, Ohlrogge JB (2009) DGAT1 and PDAT1 acyltransferases have overlapping functions in Arabidopsis triacylglycerol biosynthesis and are essential for normal pollen and seed development. Plant Cell 21:3885–3901CrossRefGoogle Scholar

Copyright information

© Springer Nature B.V. 2018

Authors and Affiliations

  • Elsinraju Devadasu
    • 1
  • Dinesh Kumar Chinthapalli
    • 1
    • 3
  • Nisha Chouhan
    • 1
  • Sai Kiran Madireddi
    • 1
  • Girish Kumar Rasineni
    • 2
  • Prabhakar Sripadi
    • 3
  • Rajagopal Subramanyam
    • 1
    Email author
  1. 1.Department of Plant Sciences, School of Life SciencesUniversity of HyderabadHyderabadIndia
  2. 2.Center for Excellence in Medical Services Pvt. Ltd.HyderabadIndia
  3. 3.Analytical Chemistry and Mass SpectrometryCSIR-Indian Institute of Chemical TechnologyHyderabadIndia

Personalised recommendations