Skip to main content
Log in

Heat shock responsiveness analysis of Athsp70b upstream region

  • Research Papers
  • Published:
Russian Journal of Plant Physiology Aims and scope Submit manuscript

Abstract

A 1431-bp upstream fragment of Athsp70b was cloned via PCR amplification and expressed in onion epidermis by particle bombardment. Furthermore, the progressive deletions of the Athsp70b upstream fragment linked to the β-glucuronidase (GUS) coding region were performed. Then, a stable GUS expression was analyzed in tobacco BY2 cells and Arabidopsis. Our present results showed that about a 500-bp region upstream ATG of Athsp70b is suitable to confer heat inducibility to the GUS reporter gene in plants and around 116 bp contain nonperfect heat-sensitive element. This promoter responds to heat, salicylic acid, and benzyladenine. GUS staining was mainly observed in the vascular tissues and root tips, implying that Athsp70b is related to water transportation.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Similar content being viewed by others

Abbreviations

BA:

benzyladenine

CRT:

C-repeat

ER:

endoplasmic reticulum

GUS:

β-glucuronidase

HSP:

heat shock protein

MS:

Murashige and Skoog nutrient medium

PEG:

polyethylene glycol

RNAi:

RNA interference

RT-PCR:

reverse transcription β polymerase chain reaction

SA:

salicylic acid

UTR:

untranslated region

References

  1. Shinmyo, A., Shoji, T., Bando, E., Nagaya, S., Nakai, Y., Kato, K., Sekine, M., and Yoshida, K., Metabolic Engineering of Cultured Tobacco Cells, Biotehnol. Bioengineer. 1998, vol. 58, pp. 329–332.

    Article  CAS  Google Scholar 

  2. Yoshida, K., Kasai, T., Garcia, M.R.C., Sawada, S., Shoji, T., Shimizu, S., Yamazaki, K., Komeda, Y., and Shinmyo, A., Heat-Inducible Expression System for a Foreign Gene in Cultured Tobacco Cells Using the HSP18.2 Promoter of Arabidopsis thaliana, Appl. Microbiol. Biotechnol., 1995, vol. 44, pp. 466–472.

    Article  PubMed  CAS  Google Scholar 

  3. Masclaux, F., Charpenteau, M., Takahashi, T., Pont-Lezica, R., and Galaud, J.P., Gene Silencing Using a Heat-Inducible RNA System in Arabidopsis, Biochem. Biophys. Res. Commun., 2004, vol. 321, pp. 364–369.

    Article  PubMed  CAS  Google Scholar 

  4. Lyznik, L.A., Hirayama, L., Rao, K.V., Abad, A., and Hodges, T.K., Heat-Inducible Expression of FLP Gene in Maize Cells, Plant J., 1995, vol. 8, pp. 177–186.

    Article  PubMed  CAS  Google Scholar 

  5. Sung, D.Y., Vierling, E., and Guy, C.L., Comprehensive Expression Profile Analysis of the Arabidopsis Hsp70 Gene Family, Plant Physiol., 2001, vol. 126, pp. 789–800.

    Article  PubMed  CAS  Google Scholar 

  6. Spena, A., Hain, R., Ziervogel, U., Saedler, H., and Schell, J., Construction of a Heat-Inducible Gene for Plants — Demonstration of Heat-Inducible Activity of the Drosophila Hsp70 Promoter in Plants, EMBO J., 1985, vol. 4, pp. 2739–2743.

    PubMed  CAS  Google Scholar 

  7. Spena, A. and Schell, J., The Expression of a Heat-Inducible Chimeric Gene in Transgenic Tobacco Plants, Mol. Gen. Genet., 1987, vol. 206, pp. 436–440.

    Article  CAS  Google Scholar 

  8. Nagata, T., Interaction of Plant Protoplast and Liposome, Methods Enzymol., 1987, vol. 148, pp. 34–39.

    Article  CAS  Google Scholar 

  9. Gynheung, An., High-Efficiency Transformation of Cultured Tobacco Cells, Plant Physiol., 1985, vol. 79, pp. 568–570.

    Article  Google Scholar 

  10. Chang, T., Chen, L., Chen, S., Cai, H., Liu, X., Xiao, G., and Zhu, Z., Transformation of Tobacco with Genes Encoding Helianthus tuberosus Agglutinin (HTA) Confers Resistance to Peach-Potato Aphid (Myzus persicae), Transgen. Res., 2003, vol. 12, pp. 607–614.

    Article  CAS  Google Scholar 

  11. Steven, J.C. and Andrew, F.B., Floral Dip: A Simplified Method for Agrobacterium-Mediated Transformation of Arabidopsis thaliana, Plant J., 1998, vol. 16, pp. 735–743.

    Article  Google Scholar 

  12. Smart, C.M., Scofield, S.R., Bevan, M.W., and Dyer, T.A., Delayed Leaf Senescence in Tobacco Plants Transformed withtmr a Gene for Cytokinin Production in Agrobacterium, Plant Cell, 1991, vol. 3, pp. 647–656.

    Article  PubMed  CAS  Google Scholar 

  13. Lichtenstein, C. and Draper, J., Genetic Engineering of Plants, DNA Cloning, Glover, D.M., Ed., Washington, IRL Press, 1985, vol. 2, pp. 67–119.

    Google Scholar 

  14. Jefferson, R.A., Assaying Chimeric Genes in Plants: The GUS Gene Fusion System, Plant Mol. Biol. Rep., 1987, vol. 5, pp. 387–405.

    Article  CAS  Google Scholar 

  15. Wooden, S.K., Li, L.J., Navarro, D., Qadri, I., Pereira, L., and Lee, A.S., Transactivation of the GRP78 Promoter by Malfolded Proteins, Glycosylation Block, and Calcium Ionophore Is Mediated through a Proximal Region Containing a CCAAT Motif Which Interacts with CTF/NF-I, Mol. Cell Biol., 1991, vol. 11, pp. 5612–5623.

    PubMed  CAS  Google Scholar 

  16. James, F.D., Christine, H.F., and Ian, M.S., Changes in Salicylic Acid and Antioxidants during Induced Thermotolerance in Mustard Seedlings, Plant Physiol., 1998, vol. 118, pp. 1455–1461.

    Article  Google Scholar 

  17. Pospisilova, J., Synkova, H., and Rulcova, J., Cytokinins and Water Stress, Biol. Plant., 2000, vol. 43, pp. 321–328.

    Article  CAS  Google Scholar 

  18. Kudoyarova, G., Valcke, R., Teplova, I., and Mustafina, A., Cytokinin Content and Transpiration of Transgenic Tobacco Plants Containing Heat-Inducible ipt-Gene as Affected by High Temperature, Biol. Plant., 1999, vol. 42,Suppl., p. S75.

    Google Scholar 

  19. Qin, L.J., Guo, X.Zh., Feng, X.Zh., Weng, L., Yan, J., Hu, X.H., and Luo, D., Cloning of LjCYC1 Gene and Nuclear Localization of LjCYC1 Protein in Lotus japonicus, J. Plant Physiol. Mol. Biol., 2004, vol. 30, pp. 523–532.

    CAS  Google Scholar 

  20. Li, J.Y., Xu, M.A., Zhou, J., and Zhang, F.Ch., Molecular Cloning and Expression Analysis of AtNHX2 Promoter, Prog. Biochem. Biophys., 2004, vol. 31, pp. 1114–1118.

    CAS  Google Scholar 

  21. Ainley, W.M. and Key, J.L., Development of a Heat Shock Inducible Expression Cassette for Plants: Characterization of Parameters for Its Use in Transient Expression Assays, Plant Mol. Biol., 1990, vol. 14, pp. 949–967.

    Article  PubMed  CAS  Google Scholar 

  22. Pelham, H.R.B., A Regulatory Upstream Promoter Element in the Drosophila HSP70 Heat-Shock Gene, Cell, 1982, vol. 30, pp. 517–528.

    Article  PubMed  CAS  Google Scholar 

  23. Wu, C., Heat Shock Transcription Factors: Structure and Regulation, Annu. Rev. Cell Dev. Biol., 1995, vol. 11, pp. 441–469.

    Article  PubMed  CAS  Google Scholar 

  24. Cohen, R.S. and Meselson, M., Inducible Transcription and Puffing in Drosophila melanogaster Transformed with Hsp70-Phage Lambda Hybrid Heat Shock Genes, Proc. Natl. Acad. Sci. USA, 1984, vol. 81, pp. 5509–5510.

    Article  PubMed  CAS  Google Scholar 

  25. Nagao, R.T. and Gurley, W.B., Use of Heat Shock Promoters to Control Gene Expression in Plants, Inducible Gene Expression, Reynolds, P.H.S., Ed., USA, 1999, pp. 97–126.

  26. DeRocher, A. and Vierling, E., Cytoplasmic HSP70 Homologues of Pea: Differential Expression in Vegetative and Embryonic Organs, Plant Mol. Biol., 1995, vol. 27, pp. 441–456.

    Article  PubMed  CAS  Google Scholar 

  27. Yu. Q.J., Du, L., Hu, Y.L., and Lin, Zh.P., Cloning a Promoter of BnPIP1 and Primarily Analyzing Its Function, Sci. China, Ser. C, 2002, vol. 36, pp. 519–527.

    Google Scholar 

  28. Prädle, R., Kloske, E., and Schöffl, F., Developmental Regulation and Tissue-Specific Differences of Heat Shock Gene Expression in Transgenic Tobacco and Arabidopsis Plants, Plant Mol. Biol., 1995, vol. 28, pp. 73–82.

    Article  Google Scholar 

Download references

Author information

Authors and Affiliations

Authors

Corresponding author

Correspondence to B. K. Kuai.

Additional information

Original Russian Text © H.L. Song, Q. Wei, J.Yu, B.K. Kuai, 2008, published in Fiziologiya Rastenii, 2008, Vol. 55, No. 5, pp. 699–709.

This text was submitted by the authors in English.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Song, H.L., Wei, Q., Yu, J. et al. Heat shock responsiveness analysis of Athsp70b upstream region. Russ J Plant Physiol 55, 629–638 (2008). https://doi.org/10.1134/S1021443708050063

Download citation

  • Received:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1134/S1021443708050063

Key words

Navigation