Document Server@UHasselt >
Research >
Research publications >

Please use this identifier to cite or link to this item: http://hdl.handle.net/1942/10009

Title: Selection of plastid- and nuclear-encoded reference genes to study the effect of altered endogenous cytokinin content on photosynthesis genes in Nicotiana tabacum
Authors: CORTLEVEN, Anne
Brenner, Wolfram G.
VALCKE, Roland
Issue Date: 2009
Publisher: SPRINGER
Citation: PHOTOSYNTHESIS RESEARCH, 102(1). p. 21-29
Abstract: Selection and use of appropriate reference genes as internal controls in real-time reverse transcription PCR (RT-PCR) assays is highly important for accurate quantification of gene expression levels. Since some photosynthetic genes are encoded in the nuclear genome and others in the chloroplast genome, we evaluated both nuclear- and plastid-encoded candidate reference genes. Six plastid-encoded candidate reference genes were derived from Arabidopsis microarray data and three plastid-and five nuclear-encoded reference genes were derived from literature. Cytokinins influence photosynthetic gene expression, so we evaluated the expression stability of the candidate reference genes in transgenic Nicotiana tabacum plants with elevated or diminished cytokinin content. We found that the most reliable strategy makes use of plastid-encoded genes for normalizing plastid photosynthetic genes and nuclear-encoded reference genes for normalizing nuclear photosynthetic genes. Compared to the use of nuclear reference genes only, this approach assimilates any effects on transcriptional activity of chloroplasts or number of chloroplast. The best expression stabilities in Nicotiana tabacum were observed for the plastid-encoded references genes Nt-RPS3, Nt-NDHI and Nt-IN1 and for the nuclear-encoded genes Nt-ACT9, Nt-alpha TUB and Nt-SSU. These genes may be suitable for normalization of photosynthetic genes under other experimental conditions in Nicotiana tabacum, and orthologues of these genes may be suitable candidates for normalizing photosynthetic gene expression in other species.
Notes: [Cortleven, Anne; Valcke, Roland] Hasselt Univ, Lab Mol & Phys Plant Physiol, Dept SBG, Ctr Environm Sci, B-3590 Diepenbeek, Belgium. [Brenner, Wolfram G.] Max Planck Inst Mol Genet, D-14195 Berlin, Germany.
URI: http://hdl.handle.net/1942/10009
DOI: 10.1007/s11120-009-9470-y
ISI #: 000270447600003
ISSN: 0166-8595
Category: A1
Type: Journal Contribution
Validation: ecoom, 2010
Appears in Collections: Research publications

Files in This Item:

There are no files associated with this item.

Items in DSpace are protected by copyright, with all rights reserved, unless otherwise indicated.