Selection of Reliable Reference Genes for Gene Expression Analysis under Abiotic Stresses in the Desert Biomass Willow, Salix psammophila

作   者：李建波，贾会霞，韩小娇，张进，孙佩，卢孟柱，胡建军*
期刊名称：Frontiers in Plant Science
影响因子：4.495
卷 期 号：7
页     码：1505
关键词：
论文摘要：
Salix psammophila is a desert shrub willow that has extraordinary adaptation to
abiotic stresses and plays an important role in maintaining local ecosystems. Moreover,
S. psammophila is regarded as a promising biomass feedstock because of its high
biomass yields and short rotation coppice cycle. However, few suitable reference genes
(RGs) for quantitative real-time polymerase chain reaction (qRT-PCR) constrain the
study on normalization of gene expression in S. psammophila until now. Here, we
investigated the expression stabilities of 14 candidate RGs across tissue types and
under four abiotic stress treatments, including heat, cold, salt, and drought treatments.
After calculation of PCR efficiencies, three different software, NormFinder, geNorm,
and BestKeeper were employed to analyze systematically the qRT-PCR data, and the
outputs were merged by RankAggreg software. The optimal RGs selected for gene
expression analysis were EF1a (Elongation factor-1 alpha) and OTU (OTU-like cysteine
protease family protein) for different tissue types, UBC (Ubiquitin-conjugating enzyme
E2) and LTA4H (Leukotriene A-4 hydrolase homolog) for heat treatment, HIS (Histone
superfamily protein H3) and ARF2 (ADP-ribosylation factor 2) for cold treatment, OTU
and ACT7 (Actin 7) for salt treatment, UBC and LTA4H for drought treatment. The
expression of UBC, ARF2, and VHAC (V-type proton ATPase subunit C) varied the
least across tissue types and under abiotic stresses. Furthermore, the relative genes
expression profiles of one tissue-specific gene WOX1a (WUSCHEL-related homeobox
1a), and four stress-inducible genes, including Hsf-A2 (Heat shock transcription factors
A2), CBF3 (C-repeat binding factor 3), HKT1 (High-Affinity KC Transporter 1), and
GST (Glutathione S-transferase), were conducted to confirm the validity of the RGs
in this study. These results provided an important RGs application guideline for gene
expression characterization in S. psammophila.