Open Access
ARTICLE
Reference Gene Selection for Quantitative Real-Time PCR Analyses of Acer palmatum under Abiotic Stress
Lu Zhu, Qiuyue Ma, Shushun Li, Jing Wen, Kunyuan Yan, Qianzhong Li*
Institute of Leisure Agriculture, Jiangsu Academy of Agricultural Sciences, Nanjing, 210014, China
* Corresponding Author: Qianzhong Li. Email: qianzhongli@ jaas.ac.cn
Phyton-International Journal of Experimental Botany 2020, 89(2), 385-403. https://doi.org/10.32604/phyton.2020.09259
Received 27 November 2019; Accepted 12 January 2020; Issue published 22 April 2020
Abstract
Quantitative real-time reverse transcriptase PCR (qRT-PCR) technology has been extensively used to estimate gene expression levels, and the selection of appropriate reference genes for qRT-PCR analysis is critically important
for obtaining authentic normalized data.
Acer palmatum is an important colorful
leaf ornamental tree species, and reference genes suitable for normalization of the
qRT-PCR data obtained from this species have not been investigated. In this
study, the expression stability of ten candidate reference genes, namely,
Actin3,
Actin6,
Actin9,
EF1α,
PP2A,
SAMDC,
TIP41,
TUBα,
TUBβ and
UBQ10, in
two distinct tissues (leaves and roots) of
A. palmatum under four different abiotic
stress conditions (cold, heat, salt and drought) were investigated and assessed
using three statistical methods (GeNorm, NormFinder and BestKeeper). The combinations of reference genes that showed stability in the different stressed samples
differed. Specifically,
Actin6,
UBQ10 and
Actin9 were the most stable reference
genes in all the samples, and
Actin3 and
Actin6 wexre stably expressed in coldstressed leaves and roots.
Actin3,
Actin6 and
UBQ10 were identified as an appropriate combination of reference genes for the analysis of heat-stressed leaves and
roots, whereas the combination of
Actin9,
UBQ10 and
Actin6 was deemed the
most suitable for the analysis of salt-stressed leaves and roots. Similarly,
Actin6
and
UBQ10 exhibited stable expression in drought-stressed leaves and roots.
Furthermore, the expression levels of CBF,
Cu/Zn-SOD and
HsfA1 were estimated
to determine the reliability of the reference genes assessed in this study. This
study revealed stable reference genes in
A. palmatum that might be used for
the normalization of qRT-PCR data obtained under various abiotic stresses.
Keywords
Cite This Article
APA Style
Zhu, L., Ma, Q., Li, S., Wen, J., Yan, K. et al. (2020). Reference gene selection for quantitative real-time PCR analyses of acer palmatum under abiotic stress. Phyton-International Journal of Experimental Botany, 89(2), 385-403. https://doi.org/10.32604/phyton.2020.09259
Vancouver Style
Zhu L, Ma Q, Li S, Wen J, Yan K, Li Q. Reference gene selection for quantitative real-time PCR analyses of acer palmatum under abiotic stress. Phyton-Int J Exp Bot. 2020;89(2):385-403 https://doi.org/10.32604/phyton.2020.09259
IEEE Style
L. Zhu, Q. Ma, S. Li, J. Wen, K. Yan, and Q. Li "Reference Gene Selection for Quantitative Real-Time PCR Analyses of Acer palmatum under Abiotic Stress," Phyton-Int. J. Exp. Bot., vol. 89, no. 2, pp. 385-403. 2020. https://doi.org/10.32604/phyton.2020.09259
Citations