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Identification of suitable grapevine reference genes for qRT-PCR derived from heterologous species

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Abstract

Identification and validation of suitable reference genes that exhibit robust transcriptional stability across many sample types is an absolute requirement of all qRT-PCR experiments. Often, however, only small numbers of reference genes, validated across limited sample types, are available for non-model species. This points to a clear need to assess and validate a wider range of potential reference genes than is currently available. We therefore looked to test and validate a large number of potential reference genes across a wide range of tissue types and treatments to determine the applicability of these reference genes for use in grapevine and other non-model plant species. Potential reference genes were selected based on stability of gene transcription in the model plant species Arabidopsis or due to their common use in the grapevine community. The selected reference genes were analyzed across two datasets consisting of a range of either ‘Sauvignon blanc’ or ‘Pinot noir’ tissues. A total of 11 potential reference genes were screened across the two datasets. Gene stability was analyzed by GeNorm, a widely used Excel application, or an ANOVA-based method developed in red clover. Both analysis methods showed that all 11 potential reference genes are stably expressed in the datasets tested, but the rankings of gene stability differed based on the datasets and analysis method used. Furthermore, the transcript stability of these genes, initially identified in Arabidopsis and now validated in grapevine, suggests applicability across a wide range of non-model plant species in addition to their utility in grapevine.

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Acknowledgments

This work is part of the New Zealand Grape and Wine Research programme, a joint investment by Plant and Food Research (PFR) and NZ Winegrowers. R.T. would like to acknowledge financial support from the Lincoln University Doctoral Scholarship scheme. We would also like to thank our colleagues at Lincoln University and PFR who have provide support throughout. In particular we would like to thank Dr. M. Trought (PFR) for guidance and intellectual support throughout this project.

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Authors and Affiliations

  1. Department of Wine, Food and Molecular Biosciences, Lincoln University, PO Box 85084, Lincoln, 7647, Canterbury, New Zealand

    Rebecca M. Tashiro, Joshua G. Philips & Christopher S. Winefield

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  1. Rebecca M. Tashiro
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  2. Joshua G. Philips
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  3. Christopher S. Winefield
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Correspondence to Christopher S. Winefield.

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The authors declare no conflict of interest.

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Communicated by S. Hohmann.

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Tashiro, R.M., Philips, J.G. & Winefield, C.S. Identification of suitable grapevine reference genes for qRT-PCR derived from heterologous species. Mol Genet Genomics 291, 483–492 (2016). https://doi.org/10.1007/s00438-015-1081-z

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  • DOI: https://doi.org/10.1007/s00438-015-1081-z

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