Open Access
ARTICLE
Internal Reference Gene Selection for Quantitative Real-Time RT-PCR Normalization in Potato Tissues
Gang Li1, Yao Zhou2, Yaqi Zhao2, Yaxue Liu2, Yuwei Ke2, Xiaoqing Jin1, Haoli Ma1,2,*
1 Department of Biological Repositories, Zhongnan Hospital of Wuhan University, Wuhan, 430071, China
2 College of Agronomy, Northwest A&F University, Yangling, Shaanxi, 712100, China
* Corresponding Author: Haoli Ma. Email:
Phyton-International Journal of Experimental Botany 2020, 89(2), 329-344. https://doi.org/10.32604/phyton.2020.08874
Received 20 October 2019; Accepted 25 February 2020; Issue published 22 April 2020
Abstract
Quantitative real-time PCR (qRT-PCR) is widely used for investigating
gene expression patterns and has many advantages, including its high sensitivity,
fidelity, and specificity. Selecting a satisfactory internal reference gene is crucial
for obtaining precise gene expression results in qRT-PCR analyses. In this study,
the transcriptomic data of 2 potato varieties were screened for housekeeping genes
with stable expression patterns. A total of 77 putative genes were selected, which
were highly and stably expressed. Then, qRT-PCR analyses were performed to
examine the expression levels of these 77 candidate reference genes in various
potato tissues, including leaves, flowers, stolons, and tubers. Gene expression
was represented by analyzing the Ct values at given threshold. Through geNorm
and NormFinder program analyses, 10 candidate genes with the most stable
expression patterns were obtained, including
RPL19,
RPS15,
RPS9,
EF1α,
TrxP1,
RPS8,
NTF,
CAM,
AACM, and
RPS28. Moreover, through the comprehensive
analyses of 4 statistical algorithms (i.e., geNorm, NormFinder, BestKeeper, and
RefFinder), results indicated that the most appropriate internal reference genes
were
RPL19 and
EF1α. The obtained stable reference genes will contribute to
future qRT-PCR analyses on potato tissue-related gene expression.
Keywords
Cite This Article
Li, G., Zhou, Y., Zhao, Y., Liu, Y., Ke, Y. et al. (2020). Internal Reference Gene Selection for Quantitative Real-Time RT-PCR Normalization in Potato Tissues.
Phyton-International Journal of Experimental Botany, 89(2), 329–344. https://doi.org/10.32604/phyton.2020.08874
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