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Normalisation of real-time RT-PCR gene expression measurements in Arabidopsis thaliana exposed to increased metal concentrations.
- Source :
-
Planta [Planta] 2008 May; Vol. 227 (6), pp. 1343-9. Date of Electronic Publication: 2008 Feb 14. - Publication Year :
- 2008
-
Abstract
- Accurate quantification by real-time RT-PCR relies on normalisation of the measured gene expression data. Normalisation with multiple reference genes is becoming the standard, but the best reference genes for gene expression studies within one organism may depend on the applied treatments or the organs and tissues studied. Ideally, reference genes should be evaluated in all experimental systems. A number of candidate reference genes for Arabidopsis have been proposed, which can be used as a starting point to evaluate their expression stability in individual experimental systems by available computer algorithms like geNorm and NormFinder. Using this approach, we identified the best three reference genes from a set of ten candidates, which included three traditional "housekeeping" genes, for normalisation of gene expression when roots and leaves of Arabidopsis thaliana are exposed to cadmium (Cd) and copper (Cu). The expression stabilities of AT5G15710 (F-box protein), AT2G28390 (SAND family protein) and AT5G08290 (mitosis protein YLS8) were the highest when considering the effect to the roots and shoots of Cd and Cu treatments. Even though the effect of Cd and excess Cu on the plants is very different, the same best reference genes were identified when considering Cd or Cu treatments separately. This suggests that these three genes may also be suitable when studying the gene expression after exposure of Arabidopsis thaliana to increased concentrations of other metals.
- Subjects :
- Cadmium pharmacology
Copper pharmacology
DNA Primers
Polymerase Chain Reaction methods
RNA, Plant genetics
Reverse Transcriptase Polymerase Chain Reaction methods
Seedlings drug effects
Seedlings genetics
Arabidopsis drug effects
Arabidopsis genetics
Gene Expression Regulation, Plant drug effects
Metals pharmacology
Subjects
Details
- Language :
- English
- ISSN :
- 0032-0935
- Volume :
- 227
- Issue :
- 6
- Database :
- MEDLINE
- Journal :
- Planta
- Publication Type :
- Academic Journal
- Accession number :
- 18273637
- Full Text :
- https://doi.org/10.1007/s00425-008-0706-4