Selection and validation of reference genes for normalization of qRT-PCR gene expression in wheat (Triticum durum L.) under drought and salt stresses

Journal of genetics

MARYAM ZABOLI^4¹  MARYAM NAZARI^5²  JAMSHIDI GOHARRIZI KIARASH^1³  MOHAMMAD MEHDI MOEMENI^1⁴  FARZANE AMIRMAHANI^3⁵  HENRY DAYTON WILDE^2⁶ 

[1] Department of Agronomy and Plant Breeding, Faculty of Agriculture, Bu-Ali Sina University, Hamedan 6517838695, Iran^5;Department of Genetic, Science and Research Branch, Islamic Azad University, Tehran 1477893855, Iran^6;Department of Plant Breeding, Yazd Branch, Islamic Azad University, Yazd 8616871967, Iran^1;Faculty of Science, Department of Chemistry, University of Birjand, Birjand 7761676334, Iran^4;Faculty of Sciences, Genetic Division, Department of Biology, University of Isfahan, Isfahan 8174673441, Iran^3;Institute of Plant Breeding, Genetics, and Genomics, University of Georgia, Athens 30602, USA^2

关键词: drought stress;    housekeeping genes;    quantitative real-time PCR;    salt stress;    transcription factors;    Triticum durum.;   

DOI  :  

学科分类：生物科学（综合）

来源: Indian Academy of Sciences

【 摘 要 】

Eight candidate housekeeping genes were examined as internal controls for normalizing expression analysis of durum wheat (Triticum durum L.) under drought and salinity stress conditions. Quantitative real-time PCR was used to analyse gene expression of multiple stress levels, plant ages (24 and 50 days old), and plant tissues (leaf and root). The algorithms BestKeeper, NormFinder, GeNorm, the delta Ct method and the RefFinder were applied to determine the stability of candidate genes. Under drought stress, the most stable reference genes were glyceraldehyde-3 phosphate, ubiquitin and βtubulin2-, whereas under salinity stress conditions, eukaryotic elongation factor 1-α, glyceraldehyde-3 phosphate and actin were identified as the most stable reference genes. Validation with stress-responsive genes NAC29 and NAC6 demonstrated that the expression level of target genes could be determined reliably with combinations of up to three of the reference genes. This is the first report on reference genes appropriate for quantification of target gene expression in T. durum under drought and salt stresses. Results of this investigation may be applicable to other Triticum species.

【 授权许可】

CC BY