Analysis of Thermotolerance in Hot Pepper Using the Antiserum Against Carrot HSP17

  • Published : 2001.01.01

Abstract

An antiserum against the carrot HSP17 (17 KDa heat shock protein) was raised using the HSP17 purified after being expressed in a recombinant E.coli in order to develop an assay system for thermotolerance in crops. The DCHsp17.7 including the coding sequence corresponding to a carrot HSP17 protein was recombined within pET-32(b) vector and achieved a maximum expression in 4 hours after an induction in E.coli. The purified DCHsp17.7 was used as an antigen to generate the corresponding antibody. The polyclonal antiserum was confirmed for it's specificity only to the low molecular weight (1mw) HSP. Besides, the possibilities to use the antiserum to interact with 1mwHSPs from other plants such as rice, cucumber, tomato, and hot pepper were examined to be plausible. To reveal any specific correlation between the amounts of 1mwHSP expressed upon HS conditions and an acquisition of thermotolerance two different approaches have been applied. first, it has been shown that only the pre-HS conditions inducing the synthesis of HSP17 allowed for the seedlings to achieve an thermotolerance and to survive the following lethal condition. Second, a western analysis using 15 different collected lines of hot peppers was performed to distinguish each other in terms of the amount of 1mwHSP. The results indicated that all 14 hot pepper lines were able to synthesize HSPs in response to an exposure to HS conditions and the amounts of the proteins synthesized at different HS temperatures were variable among the lines. There are several different patterns of 1mwHSP synthesized as a function of temperature increase observed and their correlation to physiological aspects of thermotolerance remains to be analyzed.

Keywords

References

  1. Crop Sci v.22 Adaptability of crop plants to high temperature stress Chen HH;Shen ZY;Li PH
  2. Plant Mol Biol v.16 Cloning and characterization of gene encoding low molecular weight heat shock protein from carrot Darwish KL;Wang LQ;Hwang CH;Apuya N;Zimmerman JL
  3. Plant Cell Reports v.8 Heat shock protein expression in thermotolerant and thermosensitive lines of cotton Fender SE;O'Connell MA
  4. Crop Sci v.32 Induction temperature of heat-shock protein synthesis in wheat Hendershot KL;Weng J;Nguyen HT
  5. Plant Physiol v.85 Synthesis of only two heat shock proteins is required for thermoadaptation in culture cowpea cells Heuss-Larosa K;Mayer R;Cherry JH
  6. Plant Physiol v.91 The heat shock response of carrot: Protein variations between cultured cell lines Hwang CH;Zimmerman JL
  7. Molecular characterization of the low molecular weight heat shock protein and their role in thermotolerance in carrot (Daucus carota) Hwang CH
  8. Korean J. of Plant Tissue Culture v.22 Translational control of the heat shock gene expression in carrot Hwang CH
  9. Crop Sci v.22 Adaptability of crop plants to high temperature stress Hwang HC;Shen Z-Y;Li PH
  10. Plant Physiol v.79 Presence of heat shock mRNA in field grown soybeans Kimpel JA;Key JL
  11. Plant Physiol v.90 Heat shock protein synthesis and thermal tolerance in wheat Krishnan M;Nguyen HT;Burke JJ
  12. NATO ASI Series A v.83 Multigene families of soybean heat shock proteins Key JL;Kimpel JA;Vierling E;Lin CY;L van Voten-Doting(ed.);G.S.P. Groot(ed.);T. Hall(ed.)
  13. Korean J. of Plant Tissue Culture v.27 Constitutive expression of small heat shock proteins increases thermotolerance in transgenic plant Lee B-H
  14. Plant Physiol v.74 Acquisition of thermotolerance in soybean seedlings: Synthesis and accumulation of heat shock proteins and their cellular localization Lin CY;Roberts JK;Key JL
  15. Ann Rev Biochem v.55 The heat shock response Lindquist S
  16. Dev Biol v.90 Chromatin-associated heat shock proteins of Dictystelium Loomis WF;Wheeler SA
  17. The Plant J. v.20 Modified expression of a carrot small heat shock protein gene, HSP17.7, results in increased or decreased thermo-tolerance Malik MK;Slovin JP;Hwang CH;Zimmerman JL
  18. Plant Science v.44 Synthesis of heat shock protein and acquisition of thermotolerance in high temperature tolerant and high temperature susceptible lines of sorghum Ougham HJ;Stoddart JL
  19. Physiolgia Plantarum v.82 An evaluation of gene expression during somatic embryogenesis of two temperature-sensitive carrot variants unable to complete embyro development Schnall JA;Hwang CH;Cooke TJ;Zimmerman JL
  20. Annu Rev Plant Physiology Plant Mol Biol v.42 The roles of heat shock proteins in plants Vierling E
  21. Crop Science v.32 Heat-Shock Protein gene expression in diploid wheat genotypes differing in thermal tolerance Vierling RA;Nguyen HT
  22. Science v.10 Acquired tolerance of leaves to heat Yarwood CE