He, M. and Cui, S. and Yang, X. and et al, . (2017) Selection of suitable reference genes for abiotic stress-responsive gene expression studies in peanut by real-time quantitative PCR. Electronic Journal of Biotechnology, 28. pp. 76-86.
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Abstract
Because of its strong specificity and high accuracy, real-time quantitative PCR (RT-qPCR) has been a widely used method to study the expression of genes responsive to stress. It is crucial to have a suitable set of reference genes to normalize target gene expression in peanut under different conditions using RT-qPCR. In this study, 11 candidate reference genes were selected and examined under abiotic stresses (drought, salt, heavy metal, and low temperature) and hormone (SA and ABA) conditions as well as across different organ types. Three statistical algorithms (geNorm, Norm Finder and Best Keeper) were used to evaluate the expression stabilities of reference genes, and the comprehensive rankings of gene stability were generated. Results: The results indicated that ELF1B and YLS8 were the most stable reference genes under PEG-simulated drought treatment. For high-salt treatment using NaCl, YLS8 and GAPDH were the most stable genes. Under CdCl2 treatment, UBI1 and YLS8 were suitable as stable reference genes. UBI1, ADH3, and ACTIN11 were sufficient for gene expression normalization in low-temperature experiment. All the 11 candidate reference genes showed relatively high stability under hormone treatments. For organs subset, UBI1, GAPDH, and ELF1B showed the maximum stability. UBI1 and ADH3 were the top two genes that could be used reliably in all the stress conditions assessed. Furthermore, the necessity of the reference genes screened was further confirmed by the expression pattern of AnnAhs.
| Item Type: | Article |
|---|---|
| Uncontrolled Keywords: | Abiotic stress, Drought stress, Evaluation, Expression analysis, Expression stability, Genes responsive to stress, Heavy metal, High salt, Higher-plants, Low temperature, Plant hormones |
| Author Affiliation: | North China Key Laboratory for Crop Germplasm Resources of Education Ministry, Laboratory of Crop Germplasm Resources of Hebei, Agricultural University of Hebei, Baoding 071001, People's Republic of China |
| Subjects: | Crop Improvement |
| Divisions: | Groundnut |
| Depositing User: | Mr B Krishnamurthy |
| Date Deposited: | 16 Nov 2017 03:36 |
| Last Modified: | 16 Nov 2017 03:36 |
| Official URL: | http://dx.doi.org/10.1016/j.ejbt.2017.05.004 |
| URI: | http://eprints.icrisat.ac.in/id/eprint/15163 |
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