Patterns of mitochondrial gene expression in rapeseed leaves (Brassica napus L.) at early growth stage in response to drought stress

Document Type : Research Paper


1 Former Ph.D. Student and Assistant Professor in Plant Biotechnology, Department of Plant Biotechnology, Faculty of Agricultural Sciences, University of Guilan, Rasht, Iran

2 Associate Professor, Department of Plant Biotechnology, Faculty of Agricultural Sciences, University of Guilan, Rasht, Iran

3 Assistant Professor, Department of Plant Biotechnology, Faculty of Agricultural Sciences, University of Guilan, Rasht, Iran


Drought stress adversely affects a plant’s growth and productivity. Wide ranges of molecular disorders could be caused by the production of reactive oxygen radicals. Plant cells have developed potential systems to prevent such damage by scavenging and reducing the reactive oxygen species (ROS). In this study, both the genotypes of oilseed rape-tolerant and sensitive to drought-were exposed to polyethylene glycol (PEG)-induced osmotic stress at various intervals to screen the expression of mitochondrial genes that are involved in oxidizing excessive NAD(P)H without producing adenosine triphosphate (ATP). Results showed that the maximum number of alternative oxidase 1a (AOX1a) gene expression occurred in Hyola308 after 12 hours of water stress. Meanwhile, no change was observed in other sampling times. However, in SLM046, the gene expression had gradually been increased during the stress and the maximum expression was observed after 24 hours of stress. The expression of uncoupler (UCP) gene, in SLM046, was increased during the water stress and its maximum expression was observed at eight and 24 hours after the stress. However, the maximum UCP expression in Hyola308 occurred around the 12-hour mark after the stress as an AOX gene expression. Moreover, the expression of external NADPH dehydrogenase (exNDH) was increased at the early hours of the stress in Hyola308 while the same was done during the final hours of stress in SLM046.Our results showed that high activity of the mitochondrial genes, alone or together, could also be an important factor in drought tolerance in oilseed rape crop by detoxifying the harmful effects of the ROS.


Main Subjects

  1. Bartoli, C. G., Gomez, F., Gergoff, G., Guiamét, J. J. & Puntarulo, S. (2005). Up-regulation of the mitochondrial alternative oxidase pathway enhances photosynthetic electron transport under drought conditions. Journal of experimental Botany, 56(415), 1269-1276.
  2. Begcy, K., Mariano, E. D., Mattiello, L., Nunes, A. V., Mazzafera, P., Maia, I. G. & Menossi, M. (2011). An Arabidopsis mitochondrial uncoupling protein confers tolerance to drought and salt stress in transgenic tobacco plants. PLoS One, 6(8), 237-76.
  3. Brandalise, M., Maia, I. G., Borecký, J., Vercesi, A. E. & Arruda, P. (2003). Overexpression of plant uncoupling mitochondrial protein in transgenic tobacco increases tolerance to oxidative stress. Journal of bioenergetics and biomembranes 35(3), 203-209.
  4. Edreva, A. (2005). Generation and scavenging of reactive oxygen species in chloroplasts: a submolecular approach. Agriculture, Ecosystems and Environment, 106(2), 119-133.
  5. Fernie, A. R., Carrari, F. & Sweetlove, L. J. (2004). Respiratory metabolism: glycolysis, the TCA cycle and mitochondrial electron transport. Current Opinion in Plant Biology, 7(3), 254-261.
  6. Filippou, P., Antoniou, C. & Fotopoulos, V. (2011). Effect of drought and rewatering on the cellular status and antioxidant response of Medicago truncatula plants. Plant Signaling & Behavior, 6(2), 270-277.
  7. Fu, A., Liu, H., Yu, F., Kambakam, S., Luan, S. & Rodermel, S. (2012). Alternative oxidases (AOX1a and AOX2) can functionally substitute for plastid terminal oxidase in Arabidopsis chloroplasts. The Plant Cell, 24(4), 1579-1595.
  8. Heldt, H.-W. & Piechulla, B. (2010). Plant biochemistry (Fourth Edition). London Academic Press.
  9. Hoagland, D. R. & Arnon, D. I. (1950). The water-culture method for growing plants without soil. Circular. California Agricultural Experiment Station 347. (2nd edit).
  10. Kholdebarin, B. (2004). Some physiological responses of canola (Brassica napus L.) to water deficit stress under laboratory conditions. Iranian Journal of Science and Technology (Sciences), 28(1), 43-50.
  11. Kramer, D. M., Avenson, T. J. & Edwards, G. E. (2004). Dynamic flexibility in the light reactions of photosynthesis governed by both electron and proton transfer reactions. Trends in Plant Science, 9(7), 349-357.
  12. Krauss, S., Zhang, C.-Y. & Lowell, B. B. (2005). The mitochondrial uncoupling-protein homologues. Nature Reviews Molecular Cell Biology, 6(3), 248-261.
  13. Li, C., Liang, D., Xu, R., Li, H., Zhang, Y., Qin, R., Li, L., Wei, P. & Yang, J. (2013). Overexpression of an alternative oxidase gene, OsAOX1a, improves cold tolerance in Oryza sativa L. Genetics and Molecular Research, 12, 5424-5432.
  14. Liu, Y.-J., Norberg, F. E., Szilágyi, A., De Paepe, R., Åkerlund, H.-E. & Rasmusson, A. G. (2008). The mitochondrial external NADPH dehydrogenase modulates the leaf NADPH/NADP+ ratio in transgenic Nicotiana sylvestris. Plant and Cell Physiology, 49(2), 251-263.
  15. Livak, K. J. & Schmittgen, T. D. (2001). Analysis of relative gene expression data using real-time quantitative PCR and the 2− ΔΔCT method. Methods, 25(4), 402-408.
  16. Michalecka, A. M., Agius, S. C., Møller, I. M. & Rasmusson, A. G. (2004). Identification of a mitochondrial external NADPH dehydrogenase by overexpression in transgenic Nicotiana sylvestris. The Plant Journal, 37(3), 415-425.
  17. Michalecka, A. M., Svensson, Å. S., Johansson, F. I., Agius, S. C., Johanson, U., Brennicke, A., Binder, S. & Rasmusson, A. G. (2003). Arabidopsis genes encoding mitochondrial type II NAD (P) H dehydrogenases have different evolutionary origin and show distinct responses to light. Plant Physiology, 133(2), 642-652.
  18. Michel, B. E. & Kaufmann, M. R. (1973). The osmotic potential of polyethylene glycol 6000. Plant Physiology, 51(5), 914-916.
  19. Mittler, R., Vanderauwera, S., Gollery, M. & Van Breusegem, F. (2004). Reactive oxygen gene network of plants. Trends in Plant Science, 9(10), 490-498.
  20. Moore, C. S., Cook-Johnson, R. J., Rudhe, C., Whelan, J., Day, D. A., Wiskich, J. T. & Soole, K. L. (2003). Identification of AtNDI1, an internal non-phosphorylating NAD (P) H dehydrogenase in Arabidopsis mitochondria. Plant Physiology, 133(4), 1968-1978.
  21. Nishiyama, Y., Allakhverdiev, S. I. & Murata, N. (2011). Protein synthesis is the primary target of reactive oxygen species in the photoinhibition of photosystem II. Physiologia Plantarum, 142(1), 35-46.
  22. Pastore, D., Di Pede, S. & Passarella, S. (2003). Isolated durum wheat and potato cell mitochondria oxidize externally added NADH mostly via the malate/oxaloacetate shuttle with a rate that depends on the carrier-mediated transport. Plant Physiology, 133(4), 2029-2039.
  23. Pastore, D., Fratianni, A., Di Pede, S. & Passarella, S. (2000). Effects of fatty acids, nucleotides and reactive oxygen species on durum wheat mitochondria. Febs Letters, 470(1), 88-92.
  24. Pastore, D., Stoppelli, M. C., Di Fonzo, N. & Passarella, S. (1999). The existence of the K+ channel in plant mitochondria. Journal of Biological Chemistry, 274(38), 26683-26690.
  25. Pastore, D., Trono, D., Laus, M. N., Di Fonzo, N. & Flagella, Z. (2007). Possible plant mitochondria involvement in cell adaptation to drought stress a case study: durum wheat mitochondria. Journal of Experimental Botany, 58(2), 195-210.
  26. Pastore, D., Trono, D., Laus, M. N., Di Fonzo, N. & Passarella, S. (2001). Alternative oxidase in durum wheat mitochondria. Activation by pyruvate, hydroxypyruvate and glyoxylate and physiological role. Plant and Cell Physiology, 42(12), 1373-1382.
  27. Rasmusson, A. G. & Agius, S. C. (2001). Rotenone-insensitive NAD(P)H dehydrogenases in plants: immunodetection and distribution of native proteins in mitochondria. Plant Physiology and Biochemistry, 39(12), 1057-1066.
  28. Rasmusson, A. G., Geisler, D. A. & Møller, I. M. (2008). The multiplicity of dehydrogenases in the electron transport chain of plant mitochondria. Mitochondrion, 8(1), 47-60.
  29. Rasmusson, A. G., Soole, K. L. & Elthon, T. E. (2004). Alternative NAD(P)H dehydrogenases of plant mitochondria. Annual Review of Plant Biology, 55, 23-39.
  30. Rasmusson, A. G. & Wallström, S. V. (2010). Involvement of mitochondria in the control of plant cell NAD (P) H reduction levels. Biochemical Society Transactions, 38(2), 661-666.
  31. Rhoads, D. M., Umbach, A. L., Subbaiah, C. C. & Siedow, J. N. (2006). Mitochondrial reactive oxygen species. Contribution to oxidative stress and interorganellar signaling. Plant Physiology, 141(2), 357-366.
  32. Ribas-Carbo, M., Taylor, N. L., Giles, L., Busquets, S., Finnegan, P. M., Day, D. A., Lambers, H., Medrano, H., Berry, J. A. & Flexas, J. (2005). Effects of water stress on respiration in soybean leaves. Plant Physiology, 139(1), 466-473.
  33. Schmitt, F.-J., Renger, G., Friedrich, T., Kreslavski, V. D., Zharmukhamedov, S. K., Los, D. A., Kuznetsov, V. V. & Allakhverdiev, S. I. (2014). Reactive oxygen species: re-evaluation of generation, monitoring and role in stress-signaling in phototrophic organisms. Biochimica et Biophysica Acta (BBA)-Bioenergetics, 1837(6), 835-848.
  34. Shabnam, N., Sharmila, P., Sharma, A., Strasser, R. J. & Pardha-Saradhi, P. (2015). Mitochondrial electron transport protects floating leaves of long leaf pondweed (Potamogeton nodosus Poir) against photoinhibition: comparison with submerged leaves. Photosynthesis Research, 125(1-2), 305-319.
  35. Shinozaki, K. & Yamaguchi-Shinozaki, K. (2000). Molecular responses to dehydration and low temperature: differences and cross-talk between two stress signaling pathways. Current Opinion in Plant Biology, 3(3), 217-223.
  36. Smith, A. M., Ratcliffe, R. G. & Sweetlove, L. J. (2004). Activation and function of mitochondrial uncoupling protein in plants. Journal of Biological Chemistry, 279(50), 51944-51952.
  37. Sweetlove, L. J., Lytovchenko, A., Morgan, M., Nunes-Nesi, A., Taylor, N. L., Baxter, C. J., Eickmeier, I. & Fernie, A. R. (2006). Mitochondrial uncoupling protein is required for efficient photosynthesis. Proceedings of the National Academy of Sciences, 103(51), 19587-19592.
  38. Taniguchi, M. & Miyake, H. (2012). Redox-shuttling between chloroplast and cytosol: integration of intra-chloroplast and extra-chloroplast metabolism. Current Opinion in Plant Biology, 15(3), 252-260.
  39. Türkan, İ., Bor, M., Özdemir, F. & Koca, H. (2005). Differential responses of lipid peroxidation and antioxidants in the leaves of drought-tolerant P. acutifolius Gray and drought-sensitive P. vulgaris L. subjected to polyethylene glycol mediated water stress. Plant Science, 168(1), 223-231.
  40. Valderrama, R., Corpas, F. J., Carreras, A., GÓMEZ‐RODRÍGUEZ, M. V., Chaki, M., Pedrajas, J. R., FERNÁNDEZ‐OCAÑA, A., DEL RÍO, L. A. & Barroso, J. B. (2006). The dehydrogenase‐mediated recycling of NADPH is a key antioxidant system against salt‐induced oxidative stress in olive plants. Plant, Cell & Environment, 29(7), 1449-1459.
  41. Vanlerberghe, G. C., Cvetkovska, M. & Wang, J. (2009). Is the maintenance of homeostatic mitochondrial signaling during stress a physiological role for alternative oxidase? Physiologia Plantarum, 137(4), 392-406.
  42. Vanlerberghe, G. C. & McIntosh, L. (1997). Alternative oxidase: from gene to function. Annual Review of Plant Biology, 48(1), 703-734.
  43. Vassileva, V., Simova-Stoilova, L., Demirevska, K. & Feller, U. (2009). Variety-specific response of wheat (Triticum aestivum L.) leaf mitochondria to drought stress. Journal of Plant Research, 122(4), 445-454.
  44. Vercesi, A. E., Silva, M. A. P., Leite, H. M. F., Cuccovia, I. M. & Chaimovich, H. (1995). PUMPing plants. Nature, 375(6526), 24-24.
  45. Wang, W., Vinocur, B. & Altman, A. (2003). Plant responses to drought, salinity and extreme temperatures: towards genetic engineering for stress tolerance. Planta, 218(1), 1-14.
  46. Zhu, J.-K. (2001). Plant salt tolerance. Trends in Plant Science, 6(2), 66-71.
Volume 48, Special Issue
October 2017
Pages 67-77
  • Receive Date: 01 June 2016
  • Revise Date: 20 December 2016
  • Accept Date: 21 December 2016
  • Publish Date: 23 September 2017