biologia plantarum

International journal on Plant Life established by Bohumil Němec in 1959

Biologia plantarum 61:389-393, 2017 | DOI: 10.1007/s10535-016-0665-8

Transcriptional properties of eight synthetic pathogen-inducible promoters in transgenic Arabidopsis thaliana

Z. C. Huang1, S. Peng2, H. Li1, F. H. Zeng1,*
1 School of Life Science & Technology, Lingnan Normal University, Zhanjiang, P.R. China
2 College of Environment and Life Science, Kaili University, Kaili, P.R. China

Synthetic pathogen-inducible promoters (SPIP) hold a great promise to meet the demands for a desired temporal and spatial regulation of transgenes. Four pathogen-inducible cis-elements (F-box, S-box, Gst1-box, and W-box) and the minimal cauliflower mosaic virus 35S (CaMV 35S) promoter (-46 to +8 TATA box) were used to design SPIP. Eight SPIP were synthesized and named FSGW, FSWG, GWFS, GWSF, SFGW, SFWG, WGFS, and WGSF according to the order of cis-element dimers. They were used to replace the CaMV 35S promoter in the plasmid pBI121 to control expression of the β-glucuronidase (gus) gene. The transcriptional properties of each SPIP were evaluated in homozygous T3 lines of transgenic Arabidopsis thaliana by histochemical staining gus expression and real time quantitative PCR. FSGW and FSWG had a very low basal level and a poor inducibility. The other six SPIP showed different levels of background and inducibility. Using Ralstonia solanacearum, the spores of Phytophthora capsici, and salicylic acid as inducing factors, GWSF showed the advantages of a low basal expression, rapid response, and efficient transcriptional activity in the rosette leaves of five-week-old plants. The results indicate that the permutation and combination of the cis-elements had important effects on transcriptional activities of SPIP. Synthetic pathogen-inducible promoters like GWSF are valuable because it can potentially be further improved to apply to plant genetic engineering for disease resistance.

Keywords: β-glucuronidase; pathogen-inducible cis-element; Phytophthora capsici; Ralstonia solanacearum; salicylic acid
Subjects: pathogen-inducible promoters; GUS; salicylic acid; transgenic plants; gene expression

Received: September 23, 2015; Revised: May 9, 2016; Accepted: May 24, 2016; Published: June 1, 2017  Show citation

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Huang, Z.C., Peng, S., Li, H., & Zeng, F.H. (2017). Transcriptional properties of eight synthetic pathogen-inducible promoters in transgenic Arabidopsis thaliana. Biologia plantarum, 61(2), 389-393. doi: 10.1007/s10535-016-0665-8
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    References

    1. Benfey, P.N., Ren, L., Chua, N.H.: Tissue-specific expression from CaMV 35S enhancer subdomains in early stages of plant development. - EMBO J. 9: 1677-1684, 1990. Go to original source...
    2. Cazzonelli, C.I., Velten, J.: In vivo characterization of plant promoter element interaction using synthetic promoters. - Transgenic Res. 17: 437-457, 2008. Go to original source...
    3. Chen, H., Nelson, R.S., Sherwood, J.L.: Enhanced recovery of transformants of Agrobacterium tumefaciens after freezethaw transformation and drug selection. - Biotechniques 16: 664-668, 1994.
    4. Gurr, S.J., Rushton, P.J.: Engineering plants with increased disease resistance: what are we going to express? - Trends Biotechnol. 23: 275-282, 2005. Go to original source...
    5. Heise, A., Lippok, B., Kirsch, C., Hahlbrock, K.: Two immediate-early pathogen-responsive members of the AtCMPG gene family in Arabidopsis thaliana and the Wbox-containing elicitor-response element of AtCMPG1. - Proc. nat. Acad. Sci. USA 99: 9049-9054, 2002. Go to original source...
    6. Jefferson, R.A., Kavanagh, T.A., Bevan, M.W.: GUS fusions: β-glucuronidase as a sensitive and versatile gene fusion marker in higher plants. - EMBO J. 6: 3901-3907, 1987. Go to original source...
    7. Kirsch, C., Takamiyawik, M., Schmelzer, E., Hahlbrock, K., Somssich, I.E.: A novel regulatory element involved in rapid activation of parsley ELI7 gene family members by fungal elicitor or pathogen infection. - Mol. Plant Pathol. 1: 243-251, 2000. Go to original source...
    8. Lehmeyer, M., Kanofsky, K., Hanko, E.K.R., Ahrendt, S., Wehrs, M., Machens, F., Hehl, R.: Functional dissection of a strong and specific microbe-associated molecular patternresponsive synthetic promoter. - Plant Biotechnol. J. 14: 61-71, 2016. Go to original source...
    9. Malnoy, M., Reynoird, J.P, Borejsza-Wysocka, E.E., Aldwinckle, H,S,: Activation of the pathogen-inducible Gst1 promoter of potato after elicitation by Venturia inaequalis and Erwinia amylovora in transgenic apple (Malus × domestica). - Transgenic Res. 15: 83-93, 2006. Go to original source...
    10. Niemeyer, J., Ruhe, J., Machens, F., Stahl, D.J., Hehl, R.: Inducible expression of p50 from TMV for increased resistance to bacterial crown gall disease in tobacco. - Plant mol. Biol. 84: 111-123, 2014. Go to original source...
    11. Pfaffl, M.W., Horgan, G.W., Dempfle, L.: Relative expression software tool (REST) for group-wise comparison and statistical analysis of relative expression results in real-time PCR. - Nucl. Acids Res. 30: e36, 2002. Go to original source...
    12. Roberts, K., Merkouropoulos, G., Shirsat, A. H.: Identification of promoter regions in the Arabidopsis thaliana atext1 extensin gene controlling late responses to wounding and pathogen attack. - Biol. Plant. 57: 341-350, 2013. Go to original source...
    13. Shokouhifar, F., Zamani, M. R., Motallebi, M., Mousavi, A., Malboobi, M. A.: Construction and functional analysis of pathogen-inducible synthetic promoters in Brassica napus. - Biol. Plant. 55: 689-695, 2011. Go to original source...
    14. Venter, M.: Synthetic promoters: genetic control through cis engineering. - Trends Plant Sci. 12: 118-124, 2007. Go to original source...
    15. Wang, Z., Yang, P., Fan, B., Chen, Z.: An oligo selection procedure for identification of sequence-specific DNAbinding activities associated with plant defense. - Plant J. 16: 515-522, 1998. Go to original source...
    16. Yang, Y., Li, R., Qi, M.: In vivo analysis of plant promoters and transcription factors by agroinfiltration of tobacco leaves. - Plant J. 22: 543-551, 2000. Go to original source...
    17. Zhang, X., Henriques, R., Lin, S.S., Niu, Q.W., Chua, N.H.: Agrobacterium-mediated transformation of Arabidopsis thaliana using the floral dip method. - Nat. Protocols 1: 641-646, 2006. Go to original source...
    18. Zou, C., Sun, K., Mackaluso, J.D., Seddon, A.E., Jin, R., Thomashow, M. F., Shiu, S.H.: Cis-regulatory code of stress-responsive transcription in Arabidopsis thaliana. - Proc. nat. Acad. Sci. USA 108: 14992-14997, 2011. Go to original source...

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