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Selection and validation of reliable reference genes in Gossypium raimondii

  • Original Research Paper
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Abstract

Objective

To identify reliable reference genes for gene expression analysis in Gossypium raimondii.

Results

Five different software tools, geNorm, NormFinder, BestKeeper, ReFinder and ∆Ct method were employed to analyze the qRT-PCR data systematically of 12 housekeeping genes. SAD and TUA11 showed relatively stable expression levels in all tissues (i.e. leaves, shoots, buds, and sepals). We then limited our analysis to each plant part and identified tissue-specific reference genes. Our results showed TUA11, TUB6 and EF1a, EF1a, MZA and GAPC2, MZA, GAPC2, SAD and TUA11, and UBQ and MZA were reliable reference genes in leaves, shoots, buds, and sepals, respectively.

Conclusion

Some genes were commonly identified as candidate reference genes in more than two tissue, while others were tissue-specific. Thus, our study allows choosing an appropriate control gene based on sampling for gene expression analysis.

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Abbreviations

EF-1a:

Elongation factor-1A

UBQ:

Polyubiquitin

ACT:

Actin

TUA:

α-Tubulin

TUB:

β-Tubulin

GAPC2:

Glyceraldehyde-3-phosphate dehydrogenase C-2

PTB:

Polypyrimidine tract-binding protein homolog

PP2A:

Catalytic subunit of protein phosphatase 2A (PP2A)

SAD:

Stearoyl-ACP desaturase

GM (CP):

The geometric mean of CP

AR (CP):

The arithmetic mean of CP

Min (CP) and Max (CP):

The extreme values of CP

SD (±CP):

The standard deviation of the CP

CV (% CP):

The coefficient of variance expressed as a percentage on the CP level

r:

Pearson correlation coefficient

SD:

Standard deviation

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Acknowledgments

This project is partially supported by National Natural Science Foundation of China (Grant Number: 31170263). We appreciate Faten Taki for her proofread of this manuscript.

Supplementary Table 1

Properties of twelve reference gene candidates.

Author information

Authors and Affiliations

  1. Henan Institute of Sciences and Technology, Xinxiang, 453003, Henan, People’s Republic of China

    Runrun Sun, Baohong Zhang & Qinglian Wang

  2. Department of Biology, East Carolina University, Greenville, NC, 27858, USA

    Runrun Sun, Qiuling He & Baohong Zhang

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  1. Runrun Sun
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  2. Qiuling He
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  3. Baohong Zhang
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  4. Qinglian Wang
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Correspondence to Qinglian Wang.

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Sun, R., He, Q., Zhang, B. et al. Selection and validation of reliable reference genes in Gossypium raimondii . Biotechnol Lett 37, 1483–1493 (2015). https://doi.org/10.1007/s10529-015-1810-8

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  • DOI: https://doi.org/10.1007/s10529-015-1810-8

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