Int. J. Mol. Sci. 2013, 14(12), 24338-24354; doi:10.3390/ijms141224338
Article

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

1 Key Laboratory of Tropical Plant Resource and Sustainable Use, Xishuangbanna Tropical Botanical Garden, Chinese Academy of Sciences, Menglun 666303, Yunnan, China 2 University of Chinese Academy of Sciences, Beijing 100049, China
* Author to whom correspondence should be addressed.
Received: 4 November 2013; in revised form: 27 November 2013 / Accepted: 5 December 2013 / Published: 13 December 2013
(This article belongs to the Section Biochemistry, Molecular Biology and Biophysics)
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Abstract: 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.
Keywords: physic nut; reference gene; RT-qPCR; developmental stage; abiotic stress; biofuels

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MDPI and ACS Style

Zhang, L.; He, L.-L.; Fu, Q.-T.; Xu, Z.-F. Selection of Reliable Reference Genes for Gene Expression Studies in the Biofuel Plant Jatropha curcas Using Real-Time Quantitative PCR. Int. J. Mol. Sci. 2013, 14, 24338-24354.

AMA Style

Zhang L, He L-L, Fu Q-T, Xu Z-F. Selection of Reliable Reference Genes for Gene Expression Studies in the Biofuel Plant Jatropha curcas Using Real-Time Quantitative PCR. International Journal of Molecular Sciences. 2013; 14(12):24338-24354.

Chicago/Turabian Style

Zhang, Lu; He, Liang-Liang; Fu, Qian-Tang; Xu, Zeng-Fu. 2013. "Selection of Reliable Reference Genes for Gene Expression Studies in the Biofuel Plant Jatropha curcas Using Real-Time Quantitative PCR." Int. J. Mol. Sci. 14, no. 12: 24338-24354.

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