Selection and Validation of Reference Genes for mRNA Expression by Quantitative Real-Time PCR Analysis in Neolamarckia cadamba.
Selection and Validation of Reference Genes for mRNA Expression by Quantitative Real-Time PCR Analysis in Neolamarckia cadamba.
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通过实时定量 PCR 分析 Neolamarckia cadamba mRNA 表达的参考基因的选择和验证
DOI：
10.1038/s41598-018-27633-5
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发表时间：
2018-06-18
影响因子：
4.6
通讯作者：
Peng C
中科院分区：
文献类型：
--
作者：
Huang T;Long J;Liu SW;Yang ZW;Zhu QJ;Zhao XL;Peng C
木麻黄是我国南方和东南亚重要的经济速生树种。作为未来基因表达研究的先决条件的第一步，我们已经确定并表征了一系列稳定的参考基因，这些参考基因可用作本研究中定量真实的时间PCR（qRT-PCR）表达分析的对照。使用四种不同的算法评估15个候选参考基因在不同条件下在各种组织和成熟叶片中的表达稳定性，即，geNorm、Normalware、BestKeeper和Referral。我们的研究结果表明，SAMDC是所有样品中最稳定的参考基因，成熟叶片在不同的光合周期和干旱胁迫下，而RPL 10A在各种组织中的表达最稳定。PGK和RPS 25分别被认为是成熟叶片在不同发育阶段和冷处理下的最佳参考。此外，蔗糖转运蛋白4（NcSUT 4）和9-顺式-环氧类胡萝卜素双加氧酶3（NcNCED 3）的基因表达谱用于确认候选参考基因的有效性。总的来说，我们的研究是第一个报告，以验证最佳的参考基因在各种条件下的归一化N。cadamba，将有利于未来发现该物种的基因功能。
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.
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影响因子：
5.3
作者：
Artico S;Nardeli SM;Brilhante O;Grossi-de-Sa MF;Alves-Ferreira M
通讯作者：
Alves-Ferreira M
DOI：
10.1016/j.bbrc.2014.02.004
发表时间：
2014-03-07
影响因子：
3.1
作者：
Basu, Supratim;Roychoudhury, Aryadeep;Sengupta, Dibyendu N.
通讯作者：
Sengupta, Dibyendu N.
影响因子：
--
作者：
Hu R;Fan C;Li H;Zhang Q;Fu YF
通讯作者：
Fu YF
影响因子：
5.2
作者：
Lin, Y. L.;Lai, Z. X.
通讯作者：
Lai, Z. X.
影响因子：
5.6
作者：
Hao X;Horvath DP;Chao WS;Yang Y;Wang X;Xiao B
通讯作者：
Xiao B