Photosynthesis Research

, Volume 112, Issue 1, pp 49–61 | Cite as

Characterization of photosynthesis in Arabidopsis ER-to-plastid lipid trafficking mutants

  • Ziru Li
  • Jinpeng Gao
  • Christoph Benning
  • Thomas D. SharkeyEmail author
Regular Paper


Vascular plants use two pathways to synthesize galactolipids, the predominant lipid species in chloroplasts—a prokaryotic pathway that resides entirely in the chloroplast, and a eukaryotic pathway that involves assembly in the endoplasmic reticulum. Mutants deficient in the endoplasmic reticulum pathway, trigalactosyldiacylglycerol (tgd1-1 and tgd2-1) mutants, had been previously identified with reduced contents of monogalactosyldiacylglycerol and digalactosyldiacylglycerol, and altered lipid molecular species composition. Here, we report that the altered lipid composition affected photosynthesis in lipid trafficking mutants. It was found that proton motive force as measured by electrochromic shift was reduced by ~40 % in both tgd mutants. This effect was accompanied by an increase in thylakoid conductance attributable to ATPase activity and so the rate of ATP synthesis was nearly unchanged. Thylakoid conductance to ions also increased in tgd mutants. However, gross carbon assimilation in tgd mutants as measured by gas exchange was only marginally affected. Rubisco activity, electron transport rate, and photosystem I and II oxidation status were not altered. Despite the large differences in proton motive force, responses to heat and high light stress were similar between tgd mutants and the wild type.


tgd1-1 tgd2-1 Proton motive force Electrochromic shift Galactolipids Photosynthesis 









Endoplastic reticulum




Galactolipids:galactolipids galactosyltransferase




Phosphotidic acid




Non-focusing optics spectrophotometer


Electrochromic shift


Dark-interval relaxation kinetics


Proton motive force


Photosynthetic photon flux density



We thank Dr. David Kramer for his helpful advice on using the NoFOSpec and Dr. Sean E. Weise for his technical assistance with LI-6400. This project was funded by the National Science Foundation Grants IOS-0950574 to T.D.S. and MCB-0741395 to C.B.


  1. Aronsson H, Schöttler MA, Kelly AA, Sundqvist C, Dörmann P, Karim S, Jarvis P (2008) Monogalactosyldiacylglycerol deficiency in Arabidopsis affects pigment composition in the prolamellar body and impairs thylakoid membrane energization and photoprotection in leaves. Plant Physiol 148:580–592PubMedCrossRefGoogle Scholar
  2. Avenson TJ, Cruz JA, Kanazawa A, Kramer DM (2005) Regulating the proton budget of higher plant photosynthesis. Proc Natl Acad Sci USA 102:9709–9713PubMedCrossRefGoogle Scholar
  3. Awai K, Xu C, Tamot B, Benning C (2006) A phosphatidic acid-binding protein of the chloroplast inner envelope membrane involved in lipid trafficking. Proc Natl Acad Sci USA 103:10817PubMedCrossRefGoogle Scholar
  4. Baker NR, Harbinson J, Kramer DM (2007) Determining the limitations and regulation of photosynthetic energy transduction in leaves. Plant Cell Environ 30:1107–1125PubMedCrossRefGoogle Scholar
  5. Benning C (2009) Mechanisms of lipid transport involved in organelle biogenesis in plant cells. Annu Rev Cell Dev Biol 25:71–91PubMedCrossRefGoogle Scholar
  6. Cline K, Keegstra K (1983) Galactosyltransferases involved in galactolipid biosynthesis are located in the outer membrane of pea chloroplast envelopes. Plant Physiol 71:366–372PubMedCrossRefGoogle Scholar
  7. Cruz JA, Sacksteder CA, Kanazawa A, Kramer DM (2001) Contribution of electric field (Δψ) to steady-state transthylakoid proton motive force (pmf) in vitro and in vivo. Control of pmf parsing into Δψ and ΔpH by ionic strength. Biochemistry 40:1226–1237PubMedCrossRefGoogle Scholar
  8. Dörmann P, Benning C (2002) Galactolipids rule in seed plants. Trends Plant Sci 7:112–118PubMedCrossRefGoogle Scholar
  9. Dörmann P, Hoffmann-Benning S, Balbo I, Benning C (1995) Isolation and characterization of an Arabidopsis mutant deficient in the thylakoid lipid digalactosyl diacylglycerol. Plant Cell 7:1801–1810PubMedCrossRefGoogle Scholar
  10. Dorne AJ, Block MA, Joyard J, Douce R (1982) The galactolipid:galactolipid galactosyltransferase is located on the outer surface of the outer membrane of the chloroplast envelope. FEBS Lett 145:30–34CrossRefGoogle Scholar
  11. Farquhar GD, Caemmerer S, Berry JA (1980) A biochemical model of photosynthetic CO2 assimilation in leaves of C3 species. Planta 149:78–90CrossRefGoogle Scholar
  12. Frentzen M (1986) Biosynthesis and desaturation of the different diacylglycerol moieties in higher plants. J Plant Physiol 124:193–209CrossRefGoogle Scholar
  13. Gounaris K, Barber J (1983) Monogalactosyldiacylglycerol: the most abundant polar lipid in nature. Trends Biochem Sci 8:378–381CrossRefGoogle Scholar
  14. Härtel H, Lokstein H, Dörmann P, Grimm B, Benning C (1997) Changes in the composition of the photosynthetic apparatus in the galactolipid-deficient dgd1 mutant of Arabidopsis thaliana. Plant Physiol 115:1175–1184PubMedCrossRefGoogle Scholar
  15. Härtel H, Lokstein H, Dörmann P, Trethewey RN, Benning C (1998) Photosynthetic light utilization and xanthophyll cycle activity in the galactolipid deficient dgd1 mutant of Arabidopsis thaliana. Plant Physiol Biochem 36:407–417CrossRefGoogle Scholar
  16. Hölzl G, Witt S, Gaude N, Melzer M, Schöttler MA, Dörmann P (2009) The role of diglycosyl lipids in photosynthesis and membrane lipid homeostasis in Arabidopsis. Plant Physiol 150:1147–1159PubMedCrossRefGoogle Scholar
  17. Ivanov AG, Hendrickson L, Krol M, Selstam E, Oquist G, Hurry V, Huner N (2006) Digalactosyl-diacylglycerol deficiency impairs the capacity for photosynthetic intersystem electron transport and state transitions in Arabidopsis thaliana due to photosystem I acceptor-side limitations. Plant Cell Physiol 47:1146PubMedCrossRefGoogle Scholar
  18. Jarvis P, Dörmann P, Peto CA, Lutes J, Benning C, Chory J (2000) Galactolipid deficiency and abnormal chloroplast development in the Arabidopsis MGD synthase 1 mutant. Proc Natl Acad Sci USA 97:8175–8179PubMedCrossRefGoogle Scholar
  19. Jordan P, Fromme P, Witt HT, Klukas O, Saenger W, Krausz N (2001) Three-dimensional structure of cyanobacterial photosystem I at 2.5 Å resolution. Nature 411:909–917PubMedCrossRefGoogle Scholar
  20. Joyard J, Teyssier E, Miège C, Berny-Seigneurin D, Maréchal E, Block MA, Dorne A-J, Rolland N, Ajlani G, Douce R (1998) The biochemical machinery of plastid envelope membranes. Plant Physiol 118:715–723PubMedCrossRefGoogle Scholar
  21. Kanazawa A, Kramer DM (2002) In vivo modulation of nonphotochemical exciton quenching (NPQ) by regulation of the chloroplast ATP synthase. Proc Natl Acad Sci USA 99:12789–12794PubMedCrossRefGoogle Scholar
  22. Katagiri T, Ishiyama K, Kato T, Tabata S, Kobayashi M, Shinozaki K (2005) An important role of phosphatidic acid in ABA signaling during germination in Arabidopsis thaliana. Plant J 43:107–117PubMedCrossRefGoogle Scholar
  23. Kelly AA, Froehlich JE, Dörmann P (2003) Disruption of the two digalactosyldiacylglycerol synthase genes DGD1 and DGD2 in Arabidopsis reveals the existence of an additional enzyme of galactolipid synthesis. Plant Cell 15:2694–2706PubMedCrossRefGoogle Scholar
  24. Klughammer C, Schreiber U (1994) An improved method, using saturating light pulses, for the determination of photosystem I quantum yield via P700+-absorbance changes at 830 nm. Planta 192:261–268CrossRefGoogle Scholar
  25. Krumova S, Laptenok S, Kovács L, Tóth T, van Hoek A, Garab G, van Amerongen H (2010) Digalactosyl-diacylglycerol-deficiency lowers the thermal stability of thylakoid membranes. Photosynth Res 105:229–242PubMedCrossRefGoogle Scholar
  26. Laisk A, Oja V, Rasulov B, Rämma H, Eichelmann H, Kasparova I, Pettai H, Padu E, Vapaavuori E (2002) A computer-operated routine of gas exchange and optical measurements to diagnose photosynthetic apparatus in leaves. Plant Cell Environ 25:923–943CrossRefGoogle Scholar
  27. Lichtenthaler HK (1987) Chlorophyll and carotenoids: pigments of photosynthetic membranes. Methods Enzymol 148:350–382CrossRefGoogle Scholar
  28. Liu Z, Yan H, Wang K, Kuang T, Zhang J, Gui L, An X, Chang W (2004) Crystal structure of spinach major light-harvesting complex at 2.72 A resolution. Nature 428:287–292PubMedCrossRefGoogle Scholar
  29. Loll B, Kern J, Saenger W, Zouni A, Biesiadka J (2005) Towards complete cofactor arrangement in the 3.0 Å resolution structure of photosystem II. Nature 438:1040–1044PubMedCrossRefGoogle Scholar
  30. Lu B, Benning C (2009) A 25-amino acid sequence of the Arabidopsis TGD2 protein is sufficient for specific binding of phosphatidic acid. J Biol Chem 284:17420–17427PubMedCrossRefGoogle Scholar
  31. Lu B, Xu C, Awai K, Jones AD, Benning C (2007) A small ATPase protein of Arabidopsis, TGD3, involved in chloroplast lipid import. J Biol Chem 282:35945–35953PubMedCrossRefGoogle Scholar
  32. Meinke DW, Franzmann LH, Nickle TC, Yeung EC (1994) Leafy Cotyledon mutants of Arabidopsis. Plant Cell 6:1049–1064PubMedCrossRefGoogle Scholar
  33. Mizusawa N, Wada H (2011) The role of lipids in photosystem II. Biochim Biophys Acta (BBA). doi: 10.1016/j.bbabio.2011.1004.1008 Google Scholar
  34. Moellering ER, Muthan B, Benning C (2010) Freezing tolerance in plants requires lipid remodeling at the outer chloroplast membrane. Science 330:226–228PubMedCrossRefGoogle Scholar
  35. Noctor G, Ruban AV, Horton P (1993) Modulation of Delta pH-dependent nonphotochemical quenching of chlorophyll fluorescence in spinach chloroplasts. Biochim Biophys Acta 1183:339–344CrossRefGoogle Scholar
  36. Pick U, Gounaris K, Admon A, Barber J (1984) Activation of the CF0-CF1, ATP synthase from spinach chloroplasts by chloroplast lipids. Biochim Biophys Acta 765:12–20CrossRefGoogle Scholar
  37. Pick U, Weiss M, Gounaris K, Barber J (1987) The role of different thylakoid glycolipids in the function of reconstituted chloroplast ATP synthase. Biochim Biophys Acta 891:28–39CrossRefGoogle Scholar
  38. Rossak M, Tietje C, Heinz E, Benning C (1995) Accumulation of UDP-sulfoquinovose in a sulfolipid-deficient mutant of Rhodobacter sphaeroides. J Biol Chem 270:25792–25797PubMedCrossRefGoogle Scholar
  39. Roston R, Gao J, Xu C, Benning C (2011) Arabidopsis chloroplast lipid transport protein TGD2 disrupts membranes and is part of a large complex. Plant J. doi: 10.1111/j.1365-1313X.2011.04536.x PubMedGoogle Scholar
  40. Rott M, Martins NF, Thiele W, Lein W, Bock R, Kramer DM, Schottler MA (2011) ATP synthase repression in tobacco restricts photosynthetic electron transport, CO2 assimilation, and plant growth by overacidification of the thylakoid lumen. Plant Cell 23:304–321PubMedCrossRefGoogle Scholar
  41. Roughan PG, Slack CR (1982) Cellular organization of glycerolipid metabolism. Annu Rev Plant Physiol 33:97–132CrossRefGoogle Scholar
  42. Sacksteder CA, Kramer DM (2000) Dark-interval relaxation kinetics (DIRK) of absorbance changes as a quantitative probe of steady-state electron transfer. Photosynth Res 66:145–158PubMedCrossRefGoogle Scholar
  43. Sacksteder CA, Jacoby ME, Kramer DM (2001) A portable, non-focusing optics spectrophotometer (NoFOSpec) for measurements of steady-state absorbance changes in intact plants. Photosynth Res 70:231–240PubMedCrossRefGoogle Scholar
  44. Sen A, Williams WP, Quinn PJ (1981) The structure and thermotropic properties of pure 1,2-diacylgalactosylglycerols in aqueous systems. Biochim Biophys Acta 663:380–389PubMedGoogle Scholar
  45. Sewe KU, Reich R (1977) Effect of molecular polarization on electrochromism of carotenoids. 2. Lutein-chlorophyll complexes—origin of field-indicating absorption-change at 520 nm in membranes of photosynthesis. Zeitschrift Fur Naturforschung 32c:161–171Google Scholar
  46. Sharkey TD, Bernacchi CJ, Farquhar GD, Singsaas EL (2007) Fitting photosynthetic carbon dioxide response curves for C3 leaves. Plant Cell Environ 30:1035–1040PubMedCrossRefGoogle Scholar
  47. Siebke K, Von Caemmerer S, Badger M, Furbank RT (1997) Expressing an RbcS antisense gene in transgenic Flaveria bidentis leads to an increased quantum requirement for CO2 fixed in photosystems I and II. Plant Physiol 115:1163PubMedGoogle Scholar
  48. Stroebel D, Choquet Y, Popot J-L, Picot D (2003) An atypical haem in the cytochrome b6f complex. Nature 426:413–418PubMedCrossRefGoogle Scholar
  49. Takizawa K, Cruz JA, Kanazawa A, Kramer DM (2007) The thylakoid proton motive force in vivo. Quantitative, non-invasive probes, energetics, and regulatory consequences of light-induced pmf. Biochim Biophys Acta 1767:1233–1244PubMedCrossRefGoogle Scholar
  50. Toni S (1997) Galactolipid biosynthesis genes and endosymbiosis. Trends Plant Sci 2:161–162CrossRefGoogle Scholar
  51. van Besouw A, Wintermans JFGM (1978) Galactolipid formation in chloroplast envelopes: I. Evidence for two mechanisms in galactosylation. Biochim Biophys Acta 529:44–53PubMedGoogle Scholar
  52. Wang Z, Benning C (2011) Arabidopsis thaliana polar glycerolipid profiling by thin layer chromatography (TLC) coupled with gas-liquid chromatography (GLC). J Vis Exp 49:e2518.
  53. Wang Z, Xu C, Benning C (2012) TGD4 involved in endoplasmic reticulum-to-chloroplast lipid trafficking is a phosphatidic acid binding protein. Plant J. doi: 10.1111/j.1365-1313X.2012.04900.x Google Scholar
  54. Wellburn AR, Lichtenthaler H (1984) Formulae and program to determine total carotenoids and chlorophylls a and b of leaf extracts in different solvents. Adv Photosynth Res 2:9–12Google Scholar
  55. Witt HT (1979) Energy conversion in the functional membrane of photosynthesis. Analysis by light pulse and electric pulse methods. The central role of the electric field. Biochim Biophys Acta 505:355PubMedGoogle Scholar
  56. Xu C, Fan J, Riekhof W, Froehlich JE, Benning C (2003) A permease-like protein involved in ER to thylakoid lipid transfer in Arabidopsis. EMBO J 22:2370–2379PubMedCrossRefGoogle Scholar
  57. Xu C, Fan J, Froehlich JE, Awai K, Benning C (2005) Mutation of the TGD1 chloroplast envelope protein affects phosphatidate metabolism in Arabidopsis. Plant Cell 17:3094–3110PubMedCrossRefGoogle Scholar
  58. Yamori W, Takahashi S, Makino A, Price GD, Badger MR, von Caemmerer S (2011) The roles of ATP synthase and the cytochrome b 6 /f complexes in limiting chloroplast electron transport and determining photosynthetic capacity. Plant Physiol 155:956–962PubMedCrossRefGoogle Scholar
  59. Zhang R, Sharkey TD (2009) Photosynthetic electron transport and proton flux under moderate heat stress. Photosynth Res 100:29–43PubMedCrossRefGoogle Scholar
  60. Zhang W, Qin C, Zhao J, Wang X (2004) Phospholipase Dα1-derived phosphatidic acid interacts with ABI1 phosphatase 2C and regulates abscisic acid signaling. Proc Natl Acad Sci USA 101:9508–9513PubMedCrossRefGoogle Scholar
  61. Zhang R, Cruz JA, Kramer DM, Magallanes-Lundback ME, Dellapenna D, Sharkey TD (2009) Moderate heat stress reduces the pH component of the transthylakoid proton motive force in light-adapted, intact tobacco leaves. Plant Cell Environ 32:1538–1547PubMedCrossRefGoogle Scholar
  62. Zhang R, Wise RR, Struck KR, Sharkey TD (2011) Moderate heat stress of Arabidopsis thaliana leaves causes chloroplast swelling and plastoglobule formation. Photosynth Res 105:123–134CrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media B.V. 2012

Authors and Affiliations

  • Ziru Li
    • 1
  • Jinpeng Gao
    • 1
    • 2
  • Christoph Benning
    • 1
  • Thomas D. Sharkey
    • 1
    Email author
  1. 1.Department of Biochemistry and Molecular BiologyMichigan State UniversityEast LansingUSA
  2. 2.Institute of Biological ChemistryWashington State UniversityPullmanUSA

Personalised recommendations