Selection and Validation of Reference Genes for Quantitative RT-PCR Analysis of Gene Expression in Populus trichocarpa

doi:10.3724/SP.J.1259.2013.00507

Abstract

Abstract: Quantitative RT-PCR (qRT-PCR) has been widely used in gene expression analysis because of its sensitivity, specificity, and reproducibility. Application of suitable reference genes to normalize qRT-PCR data is critical in analyzing PCR results. We analyzed the expression patterns of 7 housekeeping genes, including TUA8, TUB6, ubiquitin, GAPDH, actin, 18S rRNA and EF1α, in various tissues of greenhouse-grown Populus trichocarpa and Zn-treated in vitro plantlets. The stability of housekeeping gene expression was analyzed with use of 3 software packages, including geNorm, Norm- Finder, and BestKeeper. The genes actin, ubiquitin, EF1α and 18S rRNA were suitable reference genes for efficient normalization of qRT-PCR data, whereas TUB6 and GAPDH were not suitable for analysis of greenhouse-grown plants and Zn-treated plantlets, respectively. These findings were confirmed by comparative profiling of 4 P. trichocarpa NAC genes. This study provides useful information for reference gene selection in qRT-PCR analysis of gene expression in P. trichocarpa. It is also helpful to elucidate the function of P. trichocarpa NAC genes.

Xiaojuan Su, Baoguo Fan, Lichai Yuan, Xiuna Cui, Shanfa Lu. Selection and Validation of Reference Genes for Quantitative RT-PCR Analysis of Gene Expression in Populus trichocarpa[J]. Chinese Bulletin of Botany, 2013, 48(5): 507-518.

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URL: https://www.chinbullbotany.com/EN/10.3724/SP.J.1259.2013.00507

https://www.chinbullbotany.com/EN/Y2013/V48/I5/507

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