Environmental Science and Pollution Research

, Volume 25, Issue 9, pp 8951–8962 | Cite as

Impact of drought stress induced by polyethylene glycol on growth, water relations and cell viability of Norway spruce seedlings

  • Ilya E. Zlobin
  • Yury V. Ivanov
  • Alexander V. Kartashov
  • Vladimir V. Kuznetsov
Research Article


We investigated physiological responses of 7-week-old Norway spruce seedlings to water deficits of different intensities. Hydroponically grown seedlings were subjected to mild (−0.15 MPa), strong (−0.5 and −1.0 MPa) and extreme (−1.5 MPa) water deficit induced by polyethylene glycol 6000, and their growth parameters, water status and physiological activity were analyzed. Seedlings effectively restricted water loss under drought, and even under extreme water deficit, shoot relative water content did not fall below 85%. Water stress induced substantial decreases in the osmotic potentials of root and needle cell sap, up to 0.3–0.4 MPa under extreme water deficit, though this did not result from water loss or accumulation of K+ and Na+ ions. Seedling growth was very susceptible to water stress because of poor capacity for cell wall adjustment. Water stress injured seedling roots, as evidenced by the loss of root cell physiological activity estimated by the ability to hydrolyse fluorescein diacetate and by increased root calcium content up to 8–10-fold under extreme water stress. At the same time, root hair growth was enhanced, especially under mild water deficit, which increased the root water-absorbing capacity. In summary, seedlings of Norway spruce were characterized by high susceptibility to water stress and concurrently by pronounced ability to maintain water status. These characteristics are fully consistent with spruce confinement to moist habitats.


Picea abies Water stress Growth Cell wall adjustment Osmotic adjustment Root physiology 



We are grateful to Tatiana V. Litonova, an engineer of the Laboratory of Physiological and Molecular Mechanisms of Adaptation, Timiryazev Institute of Plant Physiology, Russian Academy of Sciences, for technical assistance.


This work was supported by the Russian Science Foundation (project No. 16-14-10224).

Compliance with ethical standards

Conflict of interest

The authors declare that they have no conflict of interest.

Supplementary material

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(GIF 154 kb)

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High resolution image (TIFF 32616 kb)


  1. Alves AAC, Setter TL (2004) Abscisic acid accumulation and osmotic adjustment in cassava under water deficit. Env Exp Bot 51(3):259–271. CrossRefGoogle Scholar
  2. Ambrose AR, Baster WL, Wong CS, Næsborg RR, Williams CB, Dawson TE (2015) Contrasting drought-response strategies in California redwoods. Tree Physiol 35(5):453–469. CrossRefGoogle Scholar
  3. Bajji M, Kinet JM, Lutts S (2001) The use of the electrolyte leakage method for assessing cell membrane stability as a water stress tolerance test in durum wheat. Plant Growth Regul 00(1):1–10. Google Scholar
  4. Bartlett MK, Scoffoni C, Sack L (2012) The determinants of leaf turgor loss point and prediction of drought tolerance of species and biomes: a global meta-analysis. Ecol Lett 15(5):393–405. CrossRefGoogle Scholar
  5. Blake TJ, Bevilacqua E, Zwiazek JJ (1991) Effects of repeated stress on turgor pressure and cell elasticity changes in black spruce seedlings. Can J For Res 21(9):1329–1333. CrossRefGoogle Scholar
  6. Blödner C, Majcherczyk A, Kües U, Polle A (2007) Early drought-induced changes to the needle proteome of Norway spruce. Tree Physiol 27(10):1423–1431. CrossRefGoogle Scholar
  7. Brunner I, Herzog C, Dawes MA, Arend M, Sperisen C (2015) How tree roots respond to drought. Front Plant Sci 29:1–16. Google Scholar
  8. Dambrine E, Carisey N, Pollier B, Granier A (1993) Effects of drought on the yellowing status and the dynamics of mineral elements in the xylem sap of declining spruce (Picea abies L.) Plant Soil 150(2):303–306. CrossRefGoogle Scholar
  9. Dichio B, Xiloyannis C, Angelopoulos K, Nuzzo V, Bufo SA, Gelano G (2003) Drought-induced variations of water relations parameters in Olea europaea. Plant Soil 257(2):381–389. CrossRefGoogle Scholar
  10. Ditmarova L, Kurjak D, Palmroth S, Kmet J, Střelcová K (2009) Physiological responses of Norway spruce (Picea abies) seedlings to drought stress. Tree Physiol 30(2):205–213. CrossRefGoogle Scholar
  11. Eldhuset TD, Nagy NE, Volařik D, Børja I, Gebauer R, Yakovlev IA, Krokene P (2013) Drought affects tracheid structure, dehydrin expression, and above- and belowground growth in 5-year-old Norway spruce. Plant Soil 366(1-2):305–320. CrossRefGoogle Scholar
  12. Fan S, Blake TJ, Blumwald E (1994) The relative contribution of elastic and osmotic adjustments to turgor maintenance of woody species. Physiol Plant 90(2):408–413. CrossRefGoogle Scholar
  13. Fedotov AN, Zhigunov AV (2016) The effect of the day length on the formation of apical buds in one-year-old containerized seedlings of Scots pine and Norway spruce. Izvestia Sankt-Peterburgskoj Lesotehniceskoj Akademii 215:80–91 (in Russian with English summary). doi:
  14. Flexas J, Bota J, Galmés J, Medrano H, Ribas-Carbó M (2006) Keeping a positive carbon balance under adverse conditions: responses of photosynthesis and respiration to water stress. Physiol Plant 127(3):343–352. CrossRefGoogle Scholar
  15. González L, González-Vilar M (2001) Determination of relative water content. In: Reigosa Roger MJ (ed) Handbook of plant ecophysiology techniques, 1st edn. Springer Netherlands, pp 207–212Google Scholar
  16. Guérard N, Maillard P, Brechet C, Lieutier F, Dreyer E (2007) Do trees use reserve or newly assimilated carbon for their defense reactions? A 13C labeling approach with young scots pines inoculated with a bark-beetle associated fungus (Ophiostoma brunneo ciliatum). Ann Sci 64(6):601–608. CrossRefGoogle Scholar
  17. Harayama H, Ikeda T, Ishida A, Yamamoto S-I (2006) Seasonal variations in water relations in current-year leaves of evergreen trees with delayed greening. Tree Physiol 26(8):1025–1033. CrossRefGoogle Scholar
  18. Hartmann H, Ziegler W, Trumbore S (2013a) Lethal drought leads to reduction in nonstructural carbohydrates in Norway spruce tree roots but not in the canopy. Funct Ecol 27(2):413–427. CrossRefGoogle Scholar
  19. Hartmann H, Ziegler W, Kolle O, Trumbore S (2013b) Thirst beats hunger—declining hydration during drought prevents carbon starvation in Norway spruce saplings. New Phytol 200(2):340–349. CrossRefGoogle Scholar
  20. Hessini K, Martínez JP, Gandour M, Albouchi A, Soltani A, Abdelly C (2009) Effect of water stress on growth, osmotic adjustment, cell wall elasticity and water-use efficiency in Spartina alterniflora. Env Exp Bot 67(2):312–319. CrossRefGoogle Scholar
  21. Hsiao TC (1973) Plant responses to water stress. Ann Rev Plant Physiol 24(1):519–570. CrossRefGoogle Scholar
  22. Hsiao TC, Xu L-K (2000) Sensitivity of growth of roots versus leaves to water stress: biophysical analysis and relation to water transport. J Exp Bot 51:1595–1616. doi:, 350
  23. Ivanov YV, Kartashov AV, Ivanova AI, Savochkin YV, Kuznetsov VV (2016) Effects of copper deficiency and copper toxicity on organogenesis and some physiological and biochemical responses of Scots pine (Pinus sylvestris L.) seedlings grown in hydroculture. Environ Sci Pollut Res 23(17):17332–17344. CrossRefGoogle Scholar
  24. Lawlor DW, Cornic G (2002) Photosynthetic carbon assimilation and associated metabolism in relation to water deficits in higher plants. Plant Cell Environ 25(2):275–294. CrossRefGoogle Scholar
  25. Le Gall H, Philippe F, Domon J-M, Gillet F, Pelloux J, Rayon C (2015) Cell wall metabolism in response to abiotic stress. Plants 4(1):112–166. CrossRefGoogle Scholar
  26. López R, Aranda I, Gil L (2009) Osmotic adjustment is a significant mechanism of drought resistance in Pinus pinaster and Pinus canariensis. Forest Systems 18(2):159–166. CrossRefGoogle Scholar
  27. Lu P, Biron P, Bréda N, Granier A (1995) Water relations of adult Norway spruce (Picea abies (L) Karst) under soil drought in the Vosges mountains: water potential, stomatal conductance and transpiration. Ann Sci For 52(2):117–129. CrossRefGoogle Scholar
  28. Lu P, Biron P, Granier A, Cochard H (1996) Water relations of adult Norway spruce (Picea abies (L) Karst) under soil drought in the Vosges mountains: whole-tree hydraulic conductance, xylem embolism and water loss regulation. Ann Sci For 53(1):113–121. CrossRefGoogle Scholar
  29. Marshall JG, Dumbroff EB (1999) Turgor regulation via cell wall adjustment in white spruce. Plant Physiol 119(1):313–319. CrossRefGoogle Scholar
  30. Marshall JG, Rutledge RG, Blumwald E, Dumbroff EB (2000) Reduction in turgid water volume in jack pine, white spruce and black spruce in response to drought and paclobutrazol. Tree Physiol 20(10):701–707. CrossRefGoogle Scholar
  31. Martínez JP, Silva H, Ledent JF, Pinto M (2007) Effect of drought stress on the osmotic adjustment, cell wall elasticity and cell volume of six cultivars of common beans (Phaseolus vulgaris L.) Europ J Agronomy 26(1):30–38. CrossRefGoogle Scholar
  32. McDowell NG (2011) Mechanisms linking drought, hydraulics, carbon metabolism, and vegetation mortality. Plant Physiol 155(3):1051–1059. CrossRefGoogle Scholar
  33. McDowell N, Pockman WT, Allen CD, Breshears DD, Cobb N, Kolb T, Plaut J, Sperry J, West A, Williams DG, Yepez EA (2008) Mechanisms of plant survival and mortality during drought: why do some plants survive while others succumb to drought? New Phytol 178(4):719–739. CrossRefGoogle Scholar
  34. Meinzer FC, Grantz DA, Goldstein G and Saliendra NZ (1990) Leaf water relations and maintenance of gas exchange in coffee cultivars grown in drying soil. Plant Physiol 94:1781-1787. doi:
  35. Meinzer F, Woodruff DR, Marias DE, McCulloh KA, Sevanto S (2014) Dynamics of leaf water relations components in co-occurring iso- and anisohydric conifer species. Plant Cell Environ 37(11):2577–2586. CrossRefGoogle Scholar
  36. Michel BE, Kaufmann MR (1973) The osmotic potential of polyethylene glycol 6000. Plant Physiol 51:5914–5916. doi:, 5, 914
  37. Moore JP, Vicré-Gibouin M, Farrant JM, Driouich A (2008) Adaptations of higher plant cell walls to water loss: drought vs desiccation. Physiol Plantarum 134(2):237–245. CrossRefGoogle Scholar
  38. Mullan D, Pietragalla J (2012) Leaf relative water content. In: Pask AJD (ed) Physiological breeding II: a field guide to wheat phenotyping, 1st edn. CIMMYT, Mexico, pp 25–27Google Scholar
  39. Muller B, Pantin F, Génard M, Turc O, Freixes S, Piques M, Gibbon Y (2011) Water deficits uncouple growth from photosynthesis, increase C content, and modify the relationships between C and growth in sink organs. J Exp Bot 62(6):1715–1729. CrossRefGoogle Scholar
  40. Pérez-López U, Robredo A, Lacuesta M, Muñoz-Rueda A, Mena-Petite A (2010) Atmospheric CO2 concentration influences the contributions of osmolyte accumulation and cell wall elasticity to salt tolerance in barley cultivars. J Plant Physiol 167(1):15–22. CrossRefGoogle Scholar
  41. Persson H, Von Fircks Y, Majdi H, Nilsson LO (1995) Root distribution in a Norway spruce (Picea abies (L.) Karst.) stand subjected to drought and ammonium-sulphate application. Plant Soil 168(1):161–165. CrossRefGoogle Scholar
  42. Pravdin LF (1975) El’ evropeyskaya i el’ sibirskaya v SSSR. Nauka, Moscow [In Russian] Google Scholar
  43. Reddy VS, Reddy ASN (2004) Proteomics of calcium-signaling components in plants. Phytochemistry 65(12):1745–1776. CrossRefGoogle Scholar
  44. Rhizopoulou S (1997) Is negative turgor fallacious? Physiol Plant 99(3):505–510. CrossRefGoogle Scholar
  45. Saruyama N, Sakakura Y, Asano T, Nishiuchi T, Sasamoto H, Kodama H (2013) Quantification of metabolic activity of cultured plant cells by vital staining with fluorescein diacetate. Anal Biochem 441(1):58–62. CrossRefGoogle Scholar
  46. Segal E, Kushnir T, Mualem Y, Shani U (2008) Water uptake and hydraulics of the root hair rhizosphere. Vadoze Zone J 7(3):1027–1034. CrossRefGoogle Scholar
  47. Serraj R, Sinclair TR (2002) Osmolyte accumulation: can it really help increase crop yield under drought conditions? Plant Cell Environ 25(2):333–341. CrossRefGoogle Scholar
  48. Shabala SN, Lew RR (2002) Turgor regulation in osmotically stressed Arabidopsis epidermal root cells. Direct support for the role of inorganic ion uptake as revealed by concurrent flux and cell turgor measurements. Plant Physiol 129(1):290–299. CrossRefGoogle Scholar
  49. Tardieu F (1996) Drought perception by plants. Do cells of droughted plants experience water stress? Plant Growth Regul 20(2):93–104. CrossRefGoogle Scholar
  50. Tardieu F (2011) Any trait or trait-related allele can confer drought tolerance: just design the right drought scenario. J Exp Bot 63(1):25–31. CrossRefGoogle Scholar
  51. Viteček J, Adam V, Petřek J, Vacek J, Kizek E, Havel L (2004) Esterases as a marker for growth of BY-2 tobacco cells and early somatic embryos of the Norway spruce. PCTOC 79(2):195–201. CrossRefGoogle Scholar
  52. Zlobin IE, Kartashov AV, Shpakovski GV (2017) Different roles of glutathione in copper and zinc chelation in Brassica napus roots. Plant Physiol Bioch 118:333–341CrossRefGoogle Scholar

Copyright information

© Springer-Verlag GmbH Germany, part of Springer Nature 2018

Authors and Affiliations

  • Ilya E. Zlobin
    • 1
  • Yury V. Ivanov
    • 1
    • 2
  • Alexander V. Kartashov
    • 1
  • Vladimir V. Kuznetsov
    • 1
  1. 1.Timiryazev Institute of Plant PhysiologyRussian Academy of SciencesMoscowRussia
  2. 2.MoscowRussia

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