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International Journal of Plant Breeding and Genetics
  Year: 2011 | Volume: 5 | Issue: 3 | Page No.: 255-267
DOI: 10.3923/ijpbg.2011.255.267
 
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Identification and Validation of Internal Control Genes for Gene Expression in Wheat Leaves Infected by Strip Rust

Xiang-Yu Long, Ya-Xi Liu, Helene Rocheleau, Therese Ouellet and Guo-Yue Chen

Abstract:
Using Quantitative real-time PCR to quantify gene expression, it is vital that one or more ideal internal control genes are used to normalize the samples to be compared. 10 traditional internal control genes were selected to evaluate the stability of expression in wheat leaves infected by strip rust, by qRT-PCR, including 18S Ribosomal RNA (18S), 26S Ribosomal RNA (26S), 60S Ribosomal protein L18a-1(60S), Actin, Cyclophilin (Cyp), Elongation factor 1 alpha (EF1A), Glyceraldehyde-3-phosphate dehydrogenase (GAPDH), Histone H3 (Histone),α-Tubulin and β-Tubulin. Expression stability of internal control genes was statistically analyzed by three different mathematical algorithms, GeNorm, NormFinder and BestKeeper. Three softwares gave similar results that 60S, Cyp and EF1A were the most stable whereas 18S and 26S did not favour normalization of qRT-PCR results in those samples. Cyp and EF1A have medium RNA transcription levels, and 18S and 26S have high RNA transcription levels. Using different normalization factors, expression levels of the target gene were obviously different. This study indicated that selection of the most stable internal control genes play an important role in gene expression and 60S, Cyp and EF1A would be ideal for normalization to target gene expression in wheat leaves infected by strip rust.
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How to cite this article:

Xiang-Yu Long, Ya-Xi Liu, Helene Rocheleau, Therese Ouellet and Guo-Yue Chen, 2011. Identification and Validation of Internal Control Genes for Gene Expression in Wheat Leaves Infected by Strip Rust. International Journal of Plant Breeding and Genetics, 5: 255-267.

DOI: 10.3923/ijpbg.2011.255.267

URL: https://scialert.net/abstract/?doi=ijpbg.2011.255.267

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