, Volume 230, Issue 1–2, pp 23–30 | Cite as

Temperature-sensitive formation of chloroplast protrusions and stromules in mesophyll cells of Arabidopsis thaliana



In leaf mesophyll cells of transgenic Arabidopsis thaliana plants expressing GFP in the chloroplast, stromules (stroma-filled tubules) with a length of up to 20 μm and a diameter of about 400–600 nm are observed in cells with spaces between the chloroplasts. They appear extremely dynamic, occasionally branched or polymorphic. In order to investigate the effect of temperature on chloroplasts, we have constructed a special temperature-controlled chamber for usage with a light microscope (LM-TCC). This LM-TCC enables presetting of the temperature for investigation directly at the microscope stage with an accuracy of ±0.1 °C in a temperature range of 0 °C to +60 °C. With the LM-TCC a temperature-dependent appearance of chloroplast protrusions has been found. These structures have a considerably smaller length-to-diameter ratio than typical stromules and reach a length of 3–5 μm. At 5–15 °C (low temperatures), almost no chloroplast protrusions are observed, but they appear with increasing temperatures. At 35–45 °C (high temperatures), numerous chloroplast protrusions with a beaklike appearance extend from a single chloroplast. Interaction of stromules with other organelles has also been investigated by transmission electron microscopy. At 20 °C, transverse sections of stromules are frequently observed with a diameter of about 450 nm. A close membrane-to-membrane contact of stromules with the nucleus and mitochondria has been visualised. Golgi stacks and microbodies are found in the spatial vicinity of stromules. At 5 °C, virtually no chloroplast protrusions or stromules are observed. At 35 °C, chloroplast protrusions are present as broader thylakoid-free stroma-filled areas, resulting in an irregular chloroplast appearance.

Keywords: Arabidopsis thaliana; Photosynthesis; Stromule; Temperature; Ultrastructure. 



confocal laser scanning microscope


light microscope temperature-controlled chamber


transmission electron microscope


Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.


  1. Bourett, TM, Czymmek, KJ, Howard, RJ 1999Ultrastructure of chloroplast protuberances in rice leaves preserved by high-pressure freezingPlanta208472479CrossRefGoogle Scholar
  2. Braun, V, Buchner, O, Neuner, G 2002Thermotolerance of photosystem 2 of three alpine plant species under field conditionsPhotosynthetica40587595CrossRefGoogle Scholar
  3. Buchner, O, Neuner, G 2001Determination of heat tolerance: a new equipment for field measurementsJ Appl Bot75130137Google Scholar
  4. Buchner, O, Neuner, G 2003Variability of heat tolerance in alpine plant species measured at different altitudesArct Antarct Alp Res35411420CrossRefGoogle Scholar
  5. Falcone, DL, Ogas, JP, Sommerville, CR 2004Regulation of membrane fatty acid composition by temperature in mutants of Arabidopsis with alterations in membrane lipid compositionBMC Plant Biol417PubMedCrossRefGoogle Scholar
  6. Gray, GR, Hope, BJ, Qin, X, Taylor, BG, Whitehead, CL 2003The characterization of photoinhibition and recovery during cold acclimation in Arabidopsis thaliana using chlorophyll fluorescence imagingPhysiol Plant119365375CrossRefGoogle Scholar
  7. Gray, JC, Sullivan, JA, Hibberd, JM, Hanson, MR 2001Stromules: mobile protrusions and interconnections between plastidsPlant Biol3223233CrossRefGoogle Scholar
  8. Gunning, BES 2005Plastid stromules: video microscopy of their outgrowth, retraction, tensioning, anchoring, branching, bridging, and tip-sheddingProtoplasma2253342PubMedCrossRefGoogle Scholar
  9. Kim, K, Portis, AR,Jr 2005Temperature dependence of photosynthesis in Arabidopsis plants with modifications in Rubisco activase and membrane fluidityPlant Cell Physiol46522530PubMedCrossRefGoogle Scholar
  10. Köhler, RH, Hanson, MR 2000Plastid tubules of higher plants are tissue-specific and developmentally regulatedJ Cell Sci1138189PubMedGoogle Scholar
  11. Köhler, RH, Cao, J, Zipfel, W, Webb, WW, Hanson, MR 1997Exchange of protein molecules through connections between higher plant plastidsScience27620392042PubMedCrossRefGoogle Scholar
  12. Kwok, EY, Hanson, MR 2003Microfilaments and microtubules control the morphology and movement of non-green plastids and stromules in Nicotiana tabacum Plant J351626PubMedCrossRefGoogle Scholar
  13. Kwok, EY, Hanson, MR 2004aStromules and the dynamic nature of plastid morphologyJ Microsc214124137CrossRefGoogle Scholar
  14. Kwok, EY, Hanson, MR 2004bIn vivo analysis of interactions between GFP-labeled microfilaments and plastid stromulesBMC Plant Biol42CrossRefGoogle Scholar
  15. Kwok, EY, Hanson, MR 2004cPlastids and stromules interact with the nucleus and cell membrane in vascular plantsPlant Cell Rep23188195CrossRefGoogle Scholar
  16. Larcher, W, Wagner, J, Lütz, C 1997Effect of heat on photosynthesis, dark respiration and cellular ultrastructure of the arctic-alpine psychrophyte Ranunculus glacialis Photosynthetica34219232CrossRefGoogle Scholar
  17. Lütz, C 1987Cytology of high alpine plants II. Microbody activity in leaves of Ranunculus glacialis LCytologia52679686Google Scholar
  18. Lütz, C, Moser, W 1977On the cytology of high alpine plants I. The ultrastructure of Ranunculus glacialis LFlora1662134Google Scholar
  19. Natesan, SKA, Sullivan, JA, Gray, JC 2005Stromules: a characteristic cell-specific feature of plastid morphologyJ Exp Bot56787797PubMedCrossRefGoogle Scholar
  20. Niki, T, Yoshida, S, Sakai, A 1978Studies on chilling injury in plant cells. I. Ultrastructural changes associated with chilling injury in callus tissues of Cornus stolonifera Plant Cell Physiol19139146Google Scholar
  21. Pyke, KA, Howells, CA 2002Plastid and stromule morphogenesis in tomatoAnn Bot90559566PubMedCrossRefGoogle Scholar
  22. Ristic, Z, Ashworth, EN 1993Changes in leave ultrastructure and carbohydrates in Arabidopsis thaliana L. (Heyn) cv. Columbia during rapid cold acclimationProtoplasma172111123CrossRefGoogle Scholar
  23. Schreiber, U, Berry, JA 1977Heat-induced changes of chlorophyll fluorescence in intact leaves correlated with damage of the photosynthetic apparatusPlanta136233238CrossRefGoogle Scholar
  24. Selga, T, Selga, M 2000The synapse-like interactions between chloroplast, dictyosome, and other cell compartments during increased ethylene production in leaves of winter rye (Secale cereale L.)Photosynthetica38433441CrossRefGoogle Scholar
  25. Spencer, D, Wildman, SG 1964The incorporation of amino acids into protein by cell-free extracts from tobacco leavesBiochemistry3954959PubMedCrossRefGoogle Scholar
  26. Stefanowska, M, Kuras, M, Kacperska, A 2002Low temperature-induced modifications in cell ultrastructure and localization of phenolics in winter oilseed rape (Brassica napus L. var. olifera L.) leavesAnn Bot90637645PubMedCrossRefGoogle Scholar
  27. Talts, P, Pärnik, T, Gardeström, P, Keerberg, O 2004Respiratory acclimation in Arabidopsis thaliana leaves at low temperatureJ Plant Physiol161573579PubMedCrossRefGoogle Scholar
  28. Tirlapur, UK, Dahse, I, Reiss, B, Meurer, J, Oelmuller, R 1999Characterization of the activity of a plastid-targeted green fluorescent protein in Arabidopsis Eur J Cell Biol78233240PubMedGoogle Scholar
  29. Valvekens, D, Van Montagu, M, Van Lijsebettens, M 1988 Agrobacterium tumefaciens-mediated transformation of Arabidopsis thaliana root explants by using kanamycin selectionProc Natl Acad Sci USA8555365540PubMedCrossRefGoogle Scholar
  30. Waters, MT, Fray, RG, Pyke, KA 2004Stromule formation is dependent upon plastid size, plastid differentiation status and the density of plastids within the cellPlant J39655667PubMedCrossRefGoogle Scholar
  31. Weis, E, Berry, JA 1988

    Plants and high temperature stress

    Long, SPWoodward, FI eds. Plants and temperatureCompany of BiologistsCambridge
    Google Scholar
  32. Wesley-Smith, J 2001Freeze-substitution of dehydrated plant tissues: artefacts of aqueous fixation revisitedProtoplasma218154167PubMedCrossRefGoogle Scholar

Copyright information

© Springer-Verlag 2007

Authors and Affiliations

  • A. Holzinger
    • 1
  • O. Buchner
    • 1
  • C. Lütz
    • 1
  • M. R. Hanson
    • 2
  1. 1.Department of Physiology and Cell Physiology of Alpine Plants, Institute of BotanyUniversity of InnsbruckInnsbruck
  2. 2.Department of Molecular Biology and GeneticsCornell UniversityIthaca

Personalised recommendations