Abstract
Transient transformation or transient expression results in rapid and fleeting gene expression. This approach is often used as a first-tier screening tool for evaluation of components that affect gene expression. Here, we describe the use of particle bombardment of lima bean cotyledons with constructs containing the green fluorescent protein (gfp) coding region for evaluation of promoter components that influence gene expression. Although this approach is conceptually quite simple, this lima bean transient expression system may not work well, if our methods and notes are not carefully read and followed. Our laboratory has successfully optimized this method over the past 10 years, resulting in a transient expression system, which works like no other that we have seen.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
Similar content being viewed by others
References
Takebe I, Labib G, Melchers G (1971) Regeneration of whole plants from isolated mesophyll protoplasts of tobacco. Naturwissenschaften 58:318–320
Horsch RB, Fry JE, Hoffmann NL, Eichholtz D, Rogers SG, Fraley RT (1985) A simple and general method for transferring genes into plants. Science 227:1229–1231
Finer JJ (2016) Generation of transgenic soybean via particle bombardment of embryogenic cultures. Curr Protoc Plant Biol 1:592–603
Yang Y, Li R, Qi M (2000) In vivo analysis of plant promoters and transcription factors by agroinfiltration of tobacco leaves. Plant J 22:543–551
Ruiz MT, Voinnet O, Baulcombe DC (1998) Initiation and maintenance of virus-induced gene silencing. Plant Cell 10:937–946
Fromm M, Taylor LP, Walbot V (1985) Expression of genes transferred into monocot and dicot plant cells by electroporation. Proc Natl Acad Sci U S A 82:5824–5828
Vain P, McMullen MD, Finer JJ (1993) Osmotic treatment enhances particle bombardment-mediated transient and stable transformation of maize. Plant Cell Rep 12:84–88
Hernandez-Garcia CM, Bouchard RA, Rushton PJ, Jones ML, Chen X, Timko MP, Finer JJ (2010) High level transgenic expression of soybean (Glycine max) GmERF and Gmubi gene promoters isolated by a novel promoter analysis pipeline. BMC Plant Biol 10:237
Gunadi A, Rushton PJ, McHale LK, Gutek AH, Finer JJ (2016) Characterization of 40 soybean (Glycine max) promoters, isolated from across 5 thematic gene groups. Plant Cell Tissue Organ Cult 127:145–160
Finer JJ, Vain P, Jones MW, McMullen MD (1992) Development of the particle inflow gun for DNA delivery into plant cells. Plant Cell Rep 11:323–328
Murashige T, Skoog F (1962) A revised medium for rapid growth and bio assays with tobacco tissue cultures. Physiol Plant 15:473–497
Finer JE, Finer JJ (2007) A simple method for reducing moisture condensation on Petri dish lids. Plant Cell Tissue Organ Cult 91:299–304
Acknowledgments
Salaries and research support were provided by the United Soybean Board and by state and federal funds appropriated to the Ohio State University/Ohio Agricultural Research and Development Center. Mention of trademark or proprietary products does not constitute a guarantee or warranty of the product and also does not imply approval to the exclusion of other products that may also be suitable.
Author information
Authors and Affiliations
Corresponding author
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2019 Springer Science+Business Media, LLC, part of Springer Nature
About this protocol
Cite this protocol
Gunadi, A., Dean, E.A., Finer, J.J. (2019). Transient Transformation Using Particle Bombardment for Gene Expression Analysis. In: Kumar, S., Barone, P., Smith, M. (eds) Transgenic Plants. Methods in Molecular Biology, vol 1864. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-8778-8_5
Download citation
DOI: https://doi.org/10.1007/978-1-4939-8778-8_5
Published:
Publisher Name: Humana Press, New York, NY
Print ISBN: 978-1-4939-8777-1
Online ISBN: 978-1-4939-8778-8
eBook Packages: Springer Protocols