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

Content Provider	Springer Nature Link
Author	Tashiro, Rebecca M. Philips, Joshua G. Winefield, Christopher S.
Copyright Year	2015
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.
Starting Page	483
Ending Page	492
Page Count	10
File Format	PDF
ISSN	16174615
Journal	Molecular and General Genetics MGG
Volume Number	291
Issue Number	1
e-ISSN	16174623
Language	English
Publisher	Springer Berlin Heidelberg
Publisher Date	2015-07-01
Publisher Place	Berlin/Heidelberg
Access Restriction	One Nation One Subscription (ONOS)
Subject Keyword	Grapevine qRT-PCR Reference genes MIQE Cell Biology Biochemistry Microbial Genetics and Genomics Plant Genetics & Genomics Animal Genetics and Genomics
Content Type	Text
Resource Type	Article
Subject	Genetics Medicine Molecular Biology

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