Selection and Validation of Reference Genes for mRNA Expression by Quantitative Real-Time PCR Analysis in Neolamarckia cadamba

• Published in: Scientific reports Vol. 8; no. 1; pp. 9311 - 11
• Main Authors: Huang, Tian, Long, Jianmei, Liu, Si-Wen, Yang, Zi-Wei, Zhu, Qi-Jin, Zhao, Xiao-Lan, Peng, Changcao
• Format: Journal Article
• Language: English
• Published: London Nature Publishing Group UK 18-06-2018; Nature Publishing Group; Nature Portfolio
• Subjects: 38/22; 38/77; 38/90; 631/1647/2017; 631/449/447; Developmental stages; Dioxygenase; DNA Primers - metabolism; Drought; Economic importance; Gene expression; Gene Expression Regulation, Plant; Genes, Plant; Humanities and Social Sciences; Leaves; multidisciplinary; Neolamarckia cadamba; Plant species; Real-Time Polymerase Chain Reaction - methods; Real-Time Polymerase Chain Reaction - standards; Reference Standards; Reproducibility of Results; RNA, Messenger - genetics; RNA, Messenger - metabolism; Rubiaceae - genetics; Science; Science (multidisciplinary); Software; Sucrose; Sucrose transporter
• ISSN: 2045-2322; 2045-2322
• DOI: 10.1038/s41598-018-27633-5

Neolamarckia cadamba
is an economically-important fast-growing tree species in South China and Southeast Asia. As a prerequisite first step for future gene expression studies, we have identified and characterized a series of stable reference genes that can be used as controls for quantitative real time PCR (qRT-PCR) expression analysis in this study. The expression stability of 15 candidate reference genes in various tissues and mature leaves under different conditions was evaluated using four different algorithms, i.e., geNorm, NormFinder, BestKeeper and RefFinder. Our results showed that
SAMDC
was the most stable of the selected reference genes across the set of all samples, mature leaves at different photosynthetic cycles and under drought stress, whereas
RPL10A
had the most stable expression in various tissues.
PGK
and
RPS25
were considered the most suitable reference for mature leaves at different developmental stages and under cold treatment, respectively. Additionally, the gene expression profiles of sucrose transporter 4 (NcSUT4), and 9‐cis‐epoxycarotenoid dioxygenase 3 (NcNCED3) were used to confirm the validity of candidate reference genes. Collectively, our study is the first report to validate the optimal reference genes for normalization under various conditions in
N
.
cadamba
and will benefit the future discovery of gene function in this species.