TY - JOUR
T1 - Housekeeping gene selection for real-time RT-PCR normalization in potato during biotic and abiotic stress
AU - Nicot, Nathalie
AU - Hausman, Jean François
AU - Hoffmann, Lucien
AU - Evers, Danièle
PY - 2005/11
Y1 - 2005/11
N2 - Plant stress studies are more and more based on gene expression. The analysis of gene expression requires sensitive, precise, and reproducible measurements for specific mRNA sequences. Real-time RT-PCR is at present the most sensitive method for the detection of low abundance mRNA. To avoid bias, real-time RT-PCR is referred to one or several internal control genes, which should not fluctuate during treatments. Here, the non-regulation of seven housekeeping genes (β-tubulin, cyclophilin, actin, elongation factor 1-α (ef1α), 18S rRNA, adenine phosphoribosyl transferase (aprt), and cytoplasmic ribosomal protein L2) during biotic (late blight) and abiotic stresses (cold and salt stress) was tested on potato plants using geNorm software. Results from the three experimental conditions indicated that ef1α was the most stable among the seven tested. The expression of the other housekeeping genes tested varied upon stress. In parallel, a study of the variability of expression of hsp20.2, shown to be implicated in late blight stress, was realized. The relative quantification of the hsp20.2 gene varied according to the internal control and the number of internal controls used, thus highlighting the importance of the choice of internal controls in such experiments.
AB - Plant stress studies are more and more based on gene expression. The analysis of gene expression requires sensitive, precise, and reproducible measurements for specific mRNA sequences. Real-time RT-PCR is at present the most sensitive method for the detection of low abundance mRNA. To avoid bias, real-time RT-PCR is referred to one or several internal control genes, which should not fluctuate during treatments. Here, the non-regulation of seven housekeeping genes (β-tubulin, cyclophilin, actin, elongation factor 1-α (ef1α), 18S rRNA, adenine phosphoribosyl transferase (aprt), and cytoplasmic ribosomal protein L2) during biotic (late blight) and abiotic stresses (cold and salt stress) was tested on potato plants using geNorm software. Results from the three experimental conditions indicated that ef1α was the most stable among the seven tested. The expression of the other housekeeping genes tested varied upon stress. In parallel, a study of the variability of expression of hsp20.2, shown to be implicated in late blight stress, was realized. The relative quantification of the hsp20.2 gene varied according to the internal control and the number of internal controls used, thus highlighting the importance of the choice of internal controls in such experiments.
KW - Abiotic stress
KW - Housekeeping genes
KW - Late blight
KW - Normalization
KW - Real-time RT-PCR
KW - Solanum tuberosum
UR - http://www.scopus.com/inward/record.url?scp=27144456802&partnerID=8YFLogxK
U2 - 10.1093/jxb/eri285
DO - 10.1093/jxb/eri285
M3 - Article
C2 - 16188960
AN - SCOPUS:27144456802
SN - 0022-0957
VL - 56
SP - 2907
EP - 2914
JO - Journal of Experimental Botany
JF - Journal of Experimental Botany
IS - 421
ER -