Selection and testing of reference genes for accurate RT-qPCR in rice seedlings under iron toxicity.
Selection and testing of reference genes for accurate RT-qPCR in rice seedlings under iron toxicity.
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DOI：
10.1371/journal.pone.0193418
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发表时间：
2018
期刊：
影响因子：
3.7
通讯作者：
de Oliveira AC
中科院分区：
文献类型：
--
作者：
Santos FICD;Marini N;Santos RSD;Hoffman BSF;Alves-Ferreira M;de Oliveira AC
逆转录定量PCR（RT-qPCR）是一种灵敏度高、重复性好的基因表达谱分析技术。然而，为了获得准确的结果，它依赖于使用内源性参考基因的表达是组成型或恒定的数据归一化。尽管该技术在植物胁迫分析中得到广泛应用，但水稻（Oryza sativa L.）还没有被彻底调查。在这里，我们测试了一组候选参考基因，用于水稻在这种压力条件下。使用四种不同的方法进行检测：Normalization、BestKeeper、geNorm和比较ΔCt。为了获得可重复和可靠的结果，遵循定量实时PCR实验（MIQE）指南的最小信息发布。在地上部（P2、OsGAPDH和OsNABP）、根（OsEF-1a、P8和OsGAPDH）和根+地上部（OsNABP、OsGAPDH和P8）中发现了有效的参考基因，使得我们能够对籼粳亚种的铁毒性进行进一步可靠的研究。这里也显示了研究传统内源基因以外的基因作为标准化物的重要性。
Reverse Transcription quantitative PCR (RT-qPCR) is a technique for gene expression profiling with high sensibility and reproducibility. However, to obtain accurate results, it depends on data normalization by using endogenous reference genes whose expression is constitutive or invariable. Although the technique is widely used in plant stress analyzes, the stability of reference genes for iron toxicity in rice (Oryza sativa L.) has not been thoroughly investigated. Here, we tested a set of candidate reference genes for use in rice under this stressful condition. The test was performed using four distinct methods: NormFinder, BestKeeper, geNorm and the comparative ΔCt. To achieve reproducible and reliable results, Minimum Information for Publication of Quantitative Real-Time PCR Experiments (MIQE) guidelines were followed. Valid reference genes were found for shoot (P2, OsGAPDH and OsNABP), root (OsEF-1a, P8 and OsGAPDH) and root+shoot (OsNABP, OsGAPDH and P8) enabling us to perform further reliable studies for iron toxicity in both indica and japonica subspecies. The importance of the study of other than the traditional endogenous genes for use as normalizers is also shown here.
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影响因子：
--
作者：
Silver N;Best S;Jiang J;Thein SL
通讯作者：
Thein SL
影响因子：
2
作者：
dos Santos, Railson Schreinert;Krueger, Mariana Madruga;de Oliveira, Antonio Costa
通讯作者：
de Oliveira, Antonio Costa
影响因子：
7.3
作者：
Sasikumar, Arjun N.;Perez, Winder B.;Kinzy, Terri Goss
通讯作者：
Kinzy, Terri Goss
DOI：
10.1186/s12284-016-0104-7
发表时间：
2016-12
期刊：
Rice (New York, N.Y.)
影响因子：
--
作者：
Pabuayon IM;Yamamoto N;Trinidad JL;Longkumer T;Raorane ML;Kohli A
通讯作者：
Kohli A
影响因子：
3.7
作者：
Chandna R;Augustine R;Bisht NC
通讯作者：
Bisht NC