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Differential expression of heat shock protein and alteration in osmolyte accumulation under heat stress in wheat

  • Ranjeet R. KumarEmail author
  • Suneha Goswami
  • Sushil K. Sharma
  • Khushboo Singh
  • Kritika A. Gadpayle
  • S. D. Singh
  • Himanshu Pathak
  • Raj D. Rai
Original Article

Abstract

Heat stress causes an array of physiological, biochemical and morphological changes in plants, affecting growth and yield. Here, we report cloning of HSP90 gene of 2,323 bp from C-306 cultivar of wheat having ORF from 62 to 2,164 bp encoded for 700 amino acids. Quantitative real time expression analysis of HSP90 gene in C-306 showed 1.5, 1.2, 2.5 fold (in root), 4.5, 4.3 and 6.5 fold increase (in flag leaf) in the transcript level at pollination, milky dough and seed hardening stages. HSP90 transcript level was observed low in root as well as shoot of susceptible cultivar (PBW343) at different stages of growth. A significant difference in the fold expression of HSP90 was observed in C-306 and PBW343 against differential heat shock. An altered expression of H2O2 and decline in proline accumulation was observed in C-306 at different stages of growth. Western blot analysis revealed the presence of 5, 6 and 5 multiprotein chaperone complexes of HSP90 in the range of 95 Da to 70 KDa at pollination, milky dough and seed hardening stages. An expression of few novel isoenzymes of superoxide dismutase and catalase was observed against differential heat shock. A decrease in cell membrane stability was observed at different stages of growth in C-306 cultivar of wheat. In conclusion, we suggest that a high HSP90 transcript level along with high activities of antioxidant isoenzymes and low proline accumulation is a promising target for developing wheat genotypes with tolerance to heat stress.

Keywords

Real time quantitative PCR Wheat HSP90 Heat stress Proline Hydrogen peroxide 

Abbreviations

CAT

Catalase

SOD

Superoxide dismutase

HS

Heat stress

HSP

Heat shock protein

ROS

Reactive oxygen species

Notes

Acknowledgments

The author sincerely thanks Indian Agricultural Research Institute (IARI) and Indian Council of Agriculture Research (ICAR) for financial assistance under National Initiative for Climate Resilient Agriculture (NICRA) project in order to take up the research work.

Supplementary material

13562_2012_106_MOESM1_ESM.pdf (198 kb)
ESM Fig. 1 (PDF 197 kb)
13562_2012_106_MOESM2_ESM.pdf (23 kb)
ESM Fig. 2 (PDF 22 kb)

References

  1. Assad MT, Paulsen GM (2002) Genetic changes in resistance to environment stress by U.S. great Plains wheat cultivars. Euphytica 128:87–96CrossRefGoogle Scholar
  2. Bandurska H (1993) In vitro and In vivo effect of proline on nitrate reductase activity under osmotic stress in barley. Acta Physiol Plant 15:83–88Google Scholar
  3. Bates LS, Waldren RP, Teare ID (1973) Rapid determination of free proline for water-stress studies. Plant Soil 39:205–207CrossRefGoogle Scholar
  4. Blum A, Ebercon A (1981) Cell membrane stability as a measure of drought and heat tolerance in wheat. Crop Sci J 21:43–47CrossRefGoogle Scholar
  5. Bohnert HJ, Jensen RG (1996) Strategies for engineering water-stress tolerance in plants. Trends Biotechnol 14:89–97CrossRefGoogle Scholar
  6. Bradford MM (1976) A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein dye binding. Anal Biochem 72:248–254PubMedCrossRefGoogle Scholar
  7. Cheeseman JM (2006) Hydrogen peroxide concentrations in leaves under natural conditions. J Exp Bot 57:2435–2444PubMedCrossRefGoogle Scholar
  8. Chen S, Smith DF (1998) Hop as an adaptor in the heat shock protein 70 (Hsp70) and hsp90 chaperone machinery. J Biol Chem 273:35194–35200PubMedCrossRefGoogle Scholar
  9. Clement M, Leonhardt N, Droillard M, Reiter I, Montillet J, Genty B, Lauriere C, Nussaume L, Noel LD (2011) The cytosolic/nuclear HSC70 and HSP90 molecular chaperones are important for stomatal closure and modulate abscisic acid-dependent physiological responses in Arabidopsis. Plant Physiol. doi: 10.1104/pp. 111.174425
  10. Feder ME, Hoffman GE (1999) Heat-shock proteins, molecular chaperones, and the stress response: evolutionary and ecological physiology. Annu Rev Physiol 61:243–282PubMedCrossRefGoogle Scholar
  11. Fokar M, Blum A, Nguyen HT (1998) Heat tolerance in spring wheat. II. Grain filling. Euphytica 104:9–15CrossRefGoogle Scholar
  12. Hare PD, Cress WA (1997) Metabolic implications of stress-induced proline accumulation in plants. Plant Growth Regul 21:79–102CrossRefGoogle Scholar
  13. Hua X, Weitao LV, Lin B, Zhang M (2011) Proline accumulation is inhibitory to Arabidopsis seedlings during heat stress. Plant Physiol. doi: 10.1104/pp. 111.175810
  14. Kele Y, Oncel I (2002) Response of antioxidative defence system to temperature and water stress combinations in wheat seedlings. Plant Sci 163:783–790CrossRefGoogle Scholar
  15. Laemmli UK (1970) Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 227:680–685PubMedCrossRefGoogle Scholar
  16. Larkindale J, Knight MR (2002) Protection against heat stress-induced oxidative damage in Arabidopsis involves calcium, abscisic acid, ethylene, and salicylic acid. Plant Physiol 128:682–695PubMedCrossRefGoogle Scholar
  17. Loreto F, Velikova V (2001) Isoprene produced by leaves protects the photosynthetic apparatus against ozone damage, quenches ozone products, and reduces lipid peroxidation of cellular membranes. Plant Physiol 127:1781–1787PubMedCrossRefGoogle Scholar
  18. Mazhar H, Basha SM (2002) Effects of desiccation on peanut (Arachis hypogea L.) seed protein composition. Environ Exp Bot 47:67–75CrossRefGoogle Scholar
  19. Morita S, Kaminaka H, Masumura T, Tanaka K (1999) Induction of rice cytosolic ascorbate peroxidase mRNA by oxidative stress; the involvement of hydrogen peroxide in oxidative stress signaling. Plant Cell Physiol 40:417–422CrossRefGoogle Scholar
  20. Muessig C, Fischer S, Altmann T (2002) Brassinosteroid-regulated gene expression. Plant Physiol 129:1241–1251CrossRefGoogle Scholar
  21. Ozden M, Demirel U, Kahraman A (2009) Effects of proline on antioxidant system in leaves of grapevine (Vitis vinifera L.) exposed to oxidative stress by H2O2. Sci Hortic 119(2):163–168CrossRefGoogle Scholar
  22. Roychaudhari G, Sarath M, Zeece MJ (2003) Reversible denaturation of soyabean kunitz trypsin inhibitor. Arch Biochem Biophys 412:20–26CrossRefGoogle Scholar
  23. Roy SK, Hiyama T, Nakamoto H (1999) Purification and characterization of the 16-kDa heat-shock-responsive protein from the thermophilic cyanobacterium Synechococcus vulcanus, which is an α-crystallin-related, small heat shock protein. Eur J Biochem 262:406–416PubMedCrossRefGoogle Scholar
  24. Rucinska R, Waplak S, Gwozdz EA (1999) Free radical formation and activity of antioxidant enzymes in lupin roots exposed to lead. Plant Physiol Biochem 37:113–115CrossRefGoogle Scholar
  25. Rizhsky L, Liang H, Shuman J, Shulaev V, Davletova S, Mitler R (2004) When defense pathways collide: the response of Arabidopsis to a combination of drought and heat stress. Plant Physiol 134:1683–1696PubMedCrossRefGoogle Scholar
  26. Saadalla MM, Shanahan JF, Quick JS (1990) Heat tolerance in winter wheat: I. Hardening and genetic effects on membrane thermostability. Crop Sci 30:1243–1247CrossRefGoogle Scholar
  27. Sairam RK, Srivastava GC (2000) Induction of oxidative stress and antioxidant activity by hydrogen peroxide treatment in tolerant and susceptible wheat genotypes. Biol Plant 43:381–386CrossRefGoogle Scholar
  28. Schoffl F, Prandl R, Reindl A (1999) Regulation of the heat-shock response. Plant Physiol 117:1135–1137CrossRefGoogle Scholar
  29. Smith DF, Whitesell L, Nair SC, Chen S, Prapapanich V, Rimerman RA (1995) Progesterone receptor structure and function altered by geldanamycin, an hsp90-binding agent. Mol Cell Biol 15:6804–6812PubMedGoogle Scholar
  30. Soliman WS, Fujimori M, Tase K, Sugiyama S (2011) Oxidative stress and physiological damage under prolonged heat stress in C3 grass Lolium perenne. Grassl Sci 57:101–106CrossRefGoogle Scholar
  31. Sung DY, Kaplan F, Lee KJ, Guy CL (2003) Acquired tolerance to temperature extremes. Trends Plant Sci 8:179–187PubMedCrossRefGoogle Scholar
  32. Vierling E (1991) The roles of heat shock proteins in plants. Annu Rev Plant Physiol Plant Mol Biol 42:579–620CrossRefGoogle Scholar

Copyright information

© Society for Plant Biochemistry and Biotechnology 2012

Authors and Affiliations

  • Ranjeet R. Kumar
    • 1
    Email author
  • Suneha Goswami
    • 1
  • Sushil K. Sharma
    • 1
  • Khushboo Singh
    • 1
  • Kritika A. Gadpayle
    • 1
  • S. D. Singh
    • 2
  • Himanshu Pathak
    • 2
  • Raj D. Rai
    • 1
  1. 1.Division of BiochemistryIndian Agricultural Research InstituteNew DelhiIndia
  2. 2.Division of Environmental ScienceIndian Agricultural Research InstituteNew DelhiIndia

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