Selection of Reliable Reference Genes for Gene Expression Analysis under Abiotic Stresses in the Desert Biomass Willow, Salix psammophila.
Selection of Reliable Reference Genes for Gene Expression Analysis under Abiotic Stresses in the Desert Biomass Willow, Salix psammophila.
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荒漠生物量杨柳非生物胁迫下基因表达分析的可靠参考基因筛选
DOI：
10.3389/fpls.2016.01505
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发表时间：
2016
影响因子：
5.6
通讯作者：
Hu J
中科院分区：
文献类型：
--
作者：
Li J;Jia H;Han X;Zhang J;Sun P;Lu M;Hu J
沙柳是一种沙漠灌木柳树，对非生物胁迫具有非凡的适应能力，在维持当地生态系统方面发挥着重要作用。此外，由于其生物质产量高和矮林轮作周期短，沙沙门冬被认为是一种有前途的生物质原料。然而，迄今为止，很少有适合定量实时聚合酶链式反应（qRT-PCR）的参考基因（RG）限制了沙沙沙门氏菌基因表达标准化的研究。在这里，我们研究了 14 个候选 RG 在不同组织类型和四种非生物胁迫处理（包括热、冷、盐和干旱处理）下的表达稳定性。计算PCR效率后，使用NormFinder、geNorm和BestKeeper三种不同的软件对qRT-PCR数据进行系统分析，并通过RankAggreg软件合并输出。选择用于基因表达分析的最佳 RG 为针对不同组织类型的 EF1α（延伸因子-1 α）和 OTU（类 OTU 半胱氨酸蛋白酶家族蛋白），针对热处理的 UBC（泛素结合酶 E2）和 LTA4H（白三烯 A-4 水解酶同源物），针对冷处理的 HIS（组蛋白超家族蛋白 H3）和 ARF2（ADP-核糖基化因子 2）处理，OTU 和 ACT7（肌动蛋白 7）用于盐处理，UBC 和 LTA4H 用于干旱处理。 UBC、ARF2 和 VHAC（V 型质子 ATP 酶亚基 C）的表达在不同组织类型和非生物胁迫下变化最小。此外，还对一种组织特异性基因 WOX1a（WUSCHEL 相关同源盒 1a）和四种应激诱导基因，包括 Hsf-A2（热休克转录因子 A2）、CBF3（C 重复结合因子 3）、HKT1（高亲和力 K+ 转运蛋白 1）和 GST（谷胱甘肽 S-转移酶）的相关基因表达谱进行了验证，以确认 RG 的有效性。在这项研究中。这些结果为沙沙沙门氏菌基因表达表征提供了重要的RGs应用指南。
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 EF1α (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 K+ 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.
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DOI：
10.1083/jcb.149.5.1087
发表时间：
2000-05-29
期刊：
The Journal of cell biology
影响因子：
--
作者：
Bhamidipati A;Lewis SA;Cowan NJ
通讯作者：
Cowan NJ
影响因子：
3.7
作者：
Kong Q;Yuan J;Niu P;Xie J;Jiang W;Huang Y;Bie Z
通讯作者：
Bie Z
影响因子：
11.6
作者：
Fiume, Elisa;Fletcher, Jennifer C.
通讯作者：
Fletcher, Jennifer C.
影响因子：
2.8
作者：
Gao, Caiqiu;Wang, Yucheng;Yang, Chuanping
通讯作者：
Yang, Chuanping
影响因子：
4.8
作者：
Kull, F;Ohlson, E;Haeggström, JZ
通讯作者：
Haeggström, JZ