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The validity of a reference gene is highly dependent on the experimental conditions in green alga Ulva linza

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Abstract

Normalization based on inappropriate reference gene may lead to the reduction of the accuracy of RT-qPCR. Although determination of suitable reference genes is essential to RT-qPCR studies, reports on the evaluation of reference genes in Ulva linza, a ubiquitous green-tide forming alga, are lacking. The expression levels of ten candidate reference genes were analyzed in U. linza across different experimental treatments, and the best-ranked reference genes differed across the treatments. The most suitable reference genes were tubulin2 (TUB2) among different salinity and UV treatments. Histone 2 (H2) was stably expressed in different temperature and desiccation stress treatments. 18S rRNA exhibited better expression stability in different light intensity treatments. While all tested samples were considered, none of single gene was widely applicable as a reference gene. Moreover, using a combination of two genes as reference genes might improve the reliability of gene expression by RT-qPCR, and the combination of TUB1 and TUB2 was selected as ideal for all tested samples. The results suggest that assessing the stability of reference gene expression patterns, determining candidates, and testing their suitability are required for each experimental investigation. The results will guide the selection of reference genes for gene expression studies in U. linza.

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Acknowledgments

This work was supported by National special fund for transgenic project (2009ZX08009-019B), the Hi-Tech Research and Development Program (863) of China (2009AA10Z106), Natural Science Foundation of Shandong Province (2009ZRA02075), Qingdao Municipal Science and Technology plan project (09-2-5-8-hy, 10-4-1-13-hy) and the National Science & Technology Pillar Program (2008BAD95B11).

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Authors and Affiliations

  1. Qingdao Agricultural University, Qingdao, 266109, China

    Meitao Dong & Wenqi Wang

  2. Yellow Sea Fisheries Research Institute, Chinese Academy of Fishery Sciences, Qingdao, 266071, China

    Xiaowen Zhang, Dong Xu & Naihao Ye

  3. Shandong Peanut Research Institute, Qingdao, 266100, People’s Republic of China

    Xiaoyuan Chi

  4. Key Laboratory of Marine Bioactive substance, The First Institute of Oceanography, State Oceanic administration (SOA), Qingdao, 266061, China

    Shanli Mou & Jianfang Xu

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  1. Meitao Dong
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  2. Xiaowen Zhang
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  3. Xiaoyuan Chi
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  4. Shanli Mou
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  5. Jianfang Xu
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  6. Dong Xu
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  7. Wenqi Wang
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  8. Naihao Ye
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Corresponding author

Correspondence to Naihao Ye.

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Communicated by F.-A. Wollman.

M. Dong and X. Zhang are contributed equally to this work.

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Dong, M., Zhang, X., Chi, X. et al. The validity of a reference gene is highly dependent on the experimental conditions in green alga Ulva linza . Curr Genet 58, 13–20 (2012). https://doi.org/10.1007/s00294-011-0361-3

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  • DOI: https://doi.org/10.1007/s00294-011-0361-3

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