Plant Molecular Biology Reporter

, Volume 30, Issue 1, pp 36–45

A Novel Non-wounding Transient Expression Assay for Cereals Mediated by Agrobacterium tumefaciens

  • Surendar Reddy Dhadi
  • Aparna Deshpande
  • Wusirika Ramakrishna
Article

DOI: 10.1007/s11105-011-0314-5

Cite this article as:
Dhadi, S.R., Deshpande, A. & Ramakrishna, W. Plant Mol Biol Rep (2012) 30: 36. doi:10.1007/s11105-011-0314-5

Abstract

A novel Agrobacterium tumefaciens-mediated transient expression assay (AmTEA) was developed for young plants of different cereal species and the model dicot Arabidopsis thaliana. AmTEA was evaluated using five promoters (six constructs) and two reporter genes, gus and egfp. The constitutive 35S promoter and the promoter of the rice glutaredoxin gene showed gus and egfp expression in the cereals analyzed in the present study. A promoter for the DEAD-box RNA helicase family protein gene from Arabidopsis showed similar expression patterns of reporter genes in stable transgenic lines as well as in transient expression lines of Arabidopsis. Agrobacterium tumefaciens co-cultivation and plant incubation times were optimized using 35S and the rice expressed protein gene promoter (R2-273). The possibility of non-specific expression of the reporter genes was ruled out by using the antibiotic carbenicillin and the comparison of expression of the reporter genes driven by full-length and truncated R2-273 promoters. AmTEA considerably reduced time, space, labor, and cost requirements. Ease of use with stress treatments is another major advantage of this method. AmTEA can be automated and used for large-scale studies to decipher promoter and gene functions with the ultimate goal to enhance the performance of cereal crops against biotic and abiotic stresses.

Keywords

RiceAgrobacteriumTransient assayCereal

Supplementary material

11105_2011_314_MOESM1_ESM.ppt (212 kb)
Figure S1Organization of the vector pBGWFS7 (http://gateway.psb.ugent.be/vector/show/pBGWFS7/search/index). (PPT 211 kb)
11105_2011_314_MOESM2_ESM.ppt (58 kb)
Figure S2Flow chart enumerating important steps in AmTEA. (PPT 58 kb)
11105_2011_314_MOESM3_ESM.ppt (190 kb)
Figure S3Evaluation of wounding and non-wounding AmTEA with R2-273 promoter using rice plants. (PPT 190 kb)

Copyright information

© Springer-Verlag 2011

Authors and Affiliations

  • Surendar Reddy Dhadi
    • 1
  • Aparna Deshpande
    • 1
  • Wusirika Ramakrishna
    • 1
  1. 1.Department of Biological SciencesMichigan Technological UniversityHoughtonUSA