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Validation of housekeeping genes for gene expression studies in an ice alga Chlamydomonas during freezing acclimation

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Abstract

Antarctic ice alga Chlamydomonas sp. ICE-L can endure extreme low temperature and high salinity stress under freezing conditions. To elucidate the molecular acclimation mechanisms using gene expression analysis, the expression stabilities of ten housekeeping genes of Chlamydomonas sp. ICE-L during freezing stress were analyzed. Some discrepancies were detected in the ranking of the candidate reference genes between geNorm and NormFinder programs, but there was substantial agreement between the groups of genes with the most and the least stable expression. RPL19 was ranked as the best candidate reference genes. Pairwise variation (V) analysis indicated the combination of two reference genes was sufficient for qRT-PCR data normalization under the experimental conditions. Considering the co-regulation between RPL19 and RPL32 (the most stable gene pairs given by geNorm program), we propose that the mean data rendered by RPL19 and GAPDH (the most stable gene pairs given by NormFinder program) be used to normalize gene expression values in Chlamydomonas sp. ICE-L more accurately. The example of FAD3 gene expression calculation demonstrated the importance of selecting an appropriate category and number of reference genes to achieve an accurate and reliable normalization of gene expression during freeze acclimation in Chlamydomonas sp. ICE-L.

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Acknowledgments

This work was financially supported by a grant of National Natural Science Foundation (40876106) for Chenlin Liu.

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Authors and Affiliations

  1. The First Institute of Oceanography, Marine Ecological Center, State Oceanic Administration, Qingdao, 266061, China

    Chenlin Liu, Guangting Wu, Xiaohang Huang, Shenghao Liu & Bailin Cong

Authors
  1. Chenlin Liu
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  2. Guangting Wu
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  3. Xiaohang Huang
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  4. Shenghao Liu
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  5. Bailin Cong
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Corresponding author

Correspondence to Chenlin Liu.

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Communicated by L. Huang.

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Liu, C., Wu, G., Huang, X. et al. Validation of housekeeping genes for gene expression studies in an ice alga Chlamydomonas during freezing acclimation. Extremophiles 16, 419–425 (2012). https://doi.org/10.1007/s00792-012-0441-4

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  • DOI: https://doi.org/10.1007/s00792-012-0441-4

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