Identifications and Validations of Reference Genes for Gene Expression Data Normalization of Chenopodium album.

Sitwat Aman; Noorul Haq; Shakeel Ahmed; Samina N Shakeel

doi:10.17957/IJAB/15.0354

Author(s)

Sitwat Aman Department of Biochemistry, Faculty of Biological Sciences, Quaid-i-Azam University 45320, Islamabad, Pakistan: Institute of Molecular Biology and Biotechnology, University of Lahore 54590, Lahore, Pakistan

Noorul Haq Department of Biochemistry, Faculty of Biological Sciences, Quaid-i-Azam University 45320, Islamabad, Pakistan; 3Department of Computer Science and BioInformatics, Khushal Khan Khattak University 27200, Karak, Khyber-Pakhtunkhwa, Pakistan

Shakeel Ahmed Physics Division, Pakistan Institute of Nuclear Science and Technology, Islamabad, Pakistan

Samina N Shakeel Department of Biochemistry, Faculty of Biological Sciences, Quaid-i-Azam University 45320, Islamabad, Pakistan; Department of Biology, Faculty of Life Sciences, Dartmouth College, Hanover, NH-03755, USA

Abstract

Quantitative real-time PCR (qRT-PCR) is considered as an important technique for gene expression and validation analysis for several high-throughput proteomics and genomics data. However, reliable qRT-PCR results are dependent on appropriate reference genes used for data normalization under specific experimental condition and such information is limited in case of plants whose genome is not sequenced yet. Therefore, we sequenced and evaluated stability of seven novel internal control genes (Act, Act a, ß-tub, 18SrRNA, GAPDH, EF1a and Ubq) of Chenopodium album, an important edible plant with medicinal values and have been recently used in synthesis as well as analysis of several gold and silver nanoparticles. Statistical algorithms like geNorm and NormFinder were used and compared for the samples treated with different types of stresses including, high/low temperatures, drought, heavy metal and salt stress. ß-tubulin was revealed as the most stable genes under heat, drought and metal stress. While, 18SrRNA has shown the stable expression only under cold and salt stresses. Our data support the use of the combinations of two stable internal control genes for normalization of the gene expression. We compared the relative expression of a stress induced Chloroplast small Heat Shock Protein (Cp-sHSP) of C. album to further support our results. Together with all novel internal controls genes, this study also provides a platform for future gene expression studies of C.album using validated housekeeping genes.

Publication Details

Page(s) 761-770

DOI 10.17957/IJAB/15.0354

Published Journal: International Journal of Agriculture and Biology, Volume: 19, Issue: 4, Year: 2017

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