Transcriptome-wide identification of optimal reference genes for expression analysis of Pyropia yezoensis responses to abiotic stress.
Transcriptome-wide identification of optimal reference genes for expression analysis of Pyropia yezoensis responses to abiotic stress.
复制标题
全转录组鉴定最佳参考基因，用于条斑紫菜对非生物胁迫反应的表达分析
DOI：
10.1186/s12864-018-4643-8
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发表时间：
2018-04-13
期刊：
影响因子：
4.4
通讯作者：
Mao Y
中科院分区：
文献类型：
--
作者：
Gao D;Kong F;Sun P;Bi G;Mao Y
研究背景：条斑Pyropia yezoensis是一种海洋红藻，是研究潮间带海藻耐非生物胁迫机制的理想模式。实时定量聚合酶链反应（RT-qPCR）是分析基因表达水平最常用的方法。为了准确地量化基因表达，选择和验证稳定的参考基因是requires.Results：我们使用转录组分析数据从不同的非生物胁迫处理，以确定6个基因相对稳定的表达水平：MAP，ATP酶，CGS 1，PPK，DPE 2，和FHP。选择这6个基因和3个常规参考基因UBC、EF 1-α和eif 4A作为最佳参考基因选择的候选基因。使用五种常用的统计方法（geNorm、ΔCt方法、Normalization、BestKeeper和Reynolds）来鉴定每个参考基因的稳定性。我们的研究结果表明：MAP、UBC和FHP在所有分析条件下稳定表达; CGS 1和UBC在脱水胁迫条件下稳定表达; MAP、UBC和CGS 1在温度胁迫条件下稳定表达。
Background:Pyropia yezoensis, a marine red alga, is an ideal research model for studying the mechanisms of abiotic stress tolerance in intertidal seaweed. Real-time quantitative polymerase chain reaction (RT-qPCR) is the most commonly used method to analyze gene expression levels. To accurately quantify gene expression, selection and validation of stable reference genes is required.Results:We used transcriptome profiling data from different abiotic stress treatments to identify six genes with relatively stable expression levels: MAP, ATPase, CGS1, PPK, DPE2, and FHP. These six genes and three conventional reference genes, UBC, EF1-α, and eif4A, were chosen as candidates for optimal reference gene selection. Five common statistical approaches (geNorm, ΔCt method, NormFinder, BestKeeper, and ReFinder) were used to identify the stability of each reference gene. Our results show that: MAP, UBC, and FHP are stably expressed in all analyzed conditions; CGS1 and UBC are stably expressed under conditions of dehydration stress; and MAP, UBC, and CGS1 are stably expressed under conditions of temperature stress.Conclusion:We have identified appropriate reference genes for RT-qPCR in P. yezoensis under different abiotic stress conditions which will facilitate studies of gene expression under these conditions.
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影响因子：
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作者：
Silver N;Best S;Jiang J;Thein SL
通讯作者：
Thein SL
影响因子：
4.1
作者：
Thellin, O;Zorzi, W;Heinen, E
通讯作者：
Heinen, E
影响因子：
--
作者：
Li L;Yan Y;Xu H;Qu T;Wang B
通讯作者：
Wang B
影响因子：
--
作者：
Maccoux, Lindsey J.;Clements, Dylan N.;Day, Philip Jr
通讯作者：
Day, Philip Jr
选择并验证合适的内参基因，用于石蒜基因表达的定量实时 PCR 分析
影响因子：
5.6
作者：
Ma, Rui;Xu, Sheng;Wang, Ren
通讯作者：
Wang, Ren