Abstract
Transcriptomic analysis using quantitative reverse transcriptase polymerase chain reaction (QRT-PCR) facilitates analysis of nuclear and mitochondrial-encoded mitochondrial genes, enabling mechanisms and regulation of signaling pathways to be explored. To illustrate this technique, we use genes of the mitochondrial respiratory chain. We show that several components of the mitochondrial respiratory chain respond to stress, in particular the alternative oxidase. This chapter describes a method involving total ribonucleic acid (RNA) isolation and QRT-PCR for the detection and analysis of transcriptional changes that accompany seven commonly used chemical stresses. This methodology describes an accurate technique to determine quantitatively absolute transcript levels and a platform to facilitate comparison between responses to other stress stimuli.
This is a preview of subscription content, log in via an institution to check access.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
References
Chen, D., Toone, W. M., Mata, J., et al. (2003) Global transcriptional responses of fission yeast to environmental stress. Mol. Biol. Cell. 14, 214–229.
Gasch, A. P., Spellman, P. T., Kao, C. M., et al. (2000) Genomic expression programs in the response of yeast cells to environmental changes. Mol. Biol. Cell 11, 4241–4257.
Causton, H. C., Ren, B., Koh, S. S., et al. (2001) Remodeling of yeast genome expression in response to environmental changes. Mol. Biol. Cell 12, 323–337.
Gray, M. W., Burger, G., and Lang, B. F. (1999) Mitochondrial evolution. Science 283, 1476–1481.
Czechowski, T., Bari, R. P., Stitt, M., Scheible, W.-R., and Udvardi, M. K. (2004) Real time RT-PCR profiling of over 1400 Arabidopsis transcription factors: unprecedented sensitivity reveals novel root-and shoot-specific genes. Plant J. 38, 366–379.
Schaffer, R., Landgraf, J., Accerbi, M., Simon, V., Larson, M., and Wisman, E. (2001) Microarray analysis of diurnal and circadian-regulated genes in Arabidopsis. Plant Cell 13, 113–123.
Clifton, R., Lister, R., Parker, K. L., et al. (2005) Stress induced co-expression of alternative respiratory chain components in Arabidopsis thaliana. Plant Mol. Biol. 58, 193–212.
Sweetlove, L. J., Heazlewood, J. L., Herald, V., et al. (2002) The impact of oxidative stress on Arabidopsis mitochondria. Plant J. 32, 891–904.
Golub, T. R., Slonim, D. K., Tamayo, P., et al. (1999) Molecular classification of cancer: class discovery and class prediction by gene expression monitoring. Science 286, 531–537.
Tamayo, P., Slonim, D., Mesirov, J., et al. (1999) Interpreting patterns of gene expression with self-organizing maps: methods and application to hematopoietic differentiation. Proc. Natl. Acad. Sci. U. S. A. 96, 2907–2912.
Author information
Authors and Affiliations
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2007 Humana Press Inc., Totowa, NJ
About this protocol
Cite this protocol
Clifton, R., Whelan, J. (2007). Plant Mitochondrial Transcriptomics by Quantitative RT-PCR. In: Leister, D., Herrmann, J.M. (eds) Mitochondria. Methods in Molecular Biology™, vol 372. Humana Press. https://doi.org/10.1007/978-1-59745-365-3_36
Download citation
DOI: https://doi.org/10.1007/978-1-59745-365-3_36
Publisher Name: Humana Press
Print ISBN: 978-1-58829-667-2
Online ISBN: 978-1-59745-365-3
eBook Packages: Springer Protocols