Int J Mol Sci:徐增富等篩選出小桐子基因表達分析的內參基因

小桐子(Jatropha curcas)是較具發展潛力的生物能源植物之一。近年來，有關小桐子的基因功能研究越來越深入，但迄今為止，還沒有針對小桐子基因表達分析篩選出合適的內參基因。隨著RT-qPCR實驗技術的廣泛應用，在分析基因的表達量時選擇合適的內參基因作為參考標準顯得尤為重要。

中科院西雙版納熱帶植物園能源植物分子育種組的博士研究生張露在導師徐增富研究員的指導下，採用RT-qPCR技術並結合qBasePLUS分析軟體，從11個常用的候選內參基因中為小桐子篩選出了適合於不同實驗樣品集（共20個樣品）的內參基因組合。研究結果表明，在不同發育階段（營養生長和生殖生長）的組織樣品集中，Actin、GAPDH和EF1α為最佳的內參基因組合；在非生物逆境脅迫（脫水脅迫和冷脅迫）的組織樣品集中，Actin、GAPDH和TUB5為最佳的內參基因組合；在單獨營養生長階段的組織樣品集中，Actin和TUB8為最佳的內參基因組合；在單獨生殖生長階段的組織樣品，GAPDH和EF1α為最佳的內參基因組合；在單獨脫水脅迫的樣品集中，TUB5和TUB8為最佳的內參基因組合；在單獨冷脅迫的樣品集中，GAPDH和Actin為最佳的內參基因組合。此外，有關脫水脅迫樣品集中內參基因組合的篩選結果也在本研究中得到標準化驗證。不同的實驗樣品集應選擇相應的內參基因組合作為基因表達量分析，本研究結果對小桐子基因功能研究中的基因表達分析具有很大的參考價值。

相關研究結果以Selection of Reliable Reference Genes for Gene Expression Studies in the Biofuel Plant Jatropha curcas Using Real-Time Quantitative PCR為題，發表在國際期刊International Journal of Molecular Sciences上。（生物谷Bioon.com）

生物谷推薦的英文摘要：

Int. J. Mol. Sci       doi：10.3390/ijms141224338

Selection of Reliable Reference Genes for Gene Expression Studies in the Biofuel Plant Jatropha curcas Using Real-Time Quantitative PCR

Lu Zhang 1，2， Liang-Liang He 1， Qian-Tang Fu 1 and Zeng-Fu Xu 1，*

Jatropha curcas is a promising renewable feedstock for biodiesel and bio-jet fuel production. To study gene expression in Jatropha in different tissues throughout development and under stress conditions， we examined a total of 11 typical candidate reference genes using real-time quantitative polymerase chain reaction (RT-qPCR) analysis， which is widely used for validating transcript levels in gene expression studies. The expression stability of these candidate reference genes was assessed across a total of 20 samples， including various tissues at vegetative and reproductive stages and under desiccation and cold stress treatments. The results obtained using software qBasePLUS showed that the top-ranked reference genes differed across the sample subsets. The combination of actin， GAPDH， and EF1α would be appropriate as a reference panel for normalizing gene expression data across samples at different developmental stages； the combination of actin， GAPDH， and TUB5 should be used as a reference panel for normalizing gene expression data across samples under various abiotic stress treatments. With regard to different developmental stages， we recommend the use of actin and TUB8 for normalization at the vegetative stage and GAPDH and EF1α for normalization at the reproductive stage. For abiotic stress treatments， we recommend the use of TUB5 and TUB8 for normalization under desiccation stress and GAPDH and actin for normalization under cold stress. These results are valuable for future research on gene expression during development or under abiotic stress in Jatropha. To our knowledge， this is the first report on the stability of reference genes in Jatropha.