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ARS Home » Pacific West Area » Aberdeen, Idaho » Small Grains and Potato Germplasm Research » Research » Publications at this Location » Publication #331010

Research Project: Genetic Improvement of Barley and Oats for Enhanced Quality and Biotic Stress Resistance

Location: Small Grains and Potato Germplasm Research

Title: Candidate qRT-PCR reference genes for barley that demonstrate better stability than traditional housekeeping genes

item GINES, MICHAEL - Brigham Young University
item Baldwin, Thomas
item Rashid, Abdur
item Esvelt Klos, Kathy
item Bregitzer, Paul
item JELLEN, ERIC - Brigham Young University
item MAUGHAN, JEFF - Brigham Young University

Submitted to: Proceedings International Barley Genetics Symposium
Publication Type: Abstract Only
Publication Acceptance Date: 1/7/2016
Publication Date: 6/30/2016
Citation: Gines, M.C., Baldwin, T.T., Rashid, A., Esvelt Klos, K.L., Bregitzer, P.P., Jellen, E., Maughan, J. 2016. Candidate qRT-PCR reference genes for barley that demonstrate better stability than traditional housekeeping genes. Proceedings International Barley Genetics Symposium. p.63.

Interpretive Summary:

Technical Abstract: Gene transcript expression analysis is a useful tool for correlating gene activity with plant phenotype. For these studies, an appropriate reference gene is necessary to quantify the expression of target genes. Classic housekeeping genes have often been used for this purpose, but may not be consistently expressed across tissues or genotypes, and empirical data for such genes in barley is lacking. Publically available bioinformatic resources allow prediction of gene expression stability for candidate reference genes through in silico analysis. Using the EST profile viewer data from Unigene, we estimated the expression stability of 655 barley genes. The twenty most-stable genes as predicted by in silico analysis were evaluated in the barley cultivar ‘Conlon’ by qRT-PCR across eight tissues. The five genes with the most stable expression were identified as: superoxide dismutase [Cu-Zn], malate dehydrogenase, elongation factor 2, a 3-ketoacyl-CoA thiolase-like protein, and S-adenosylmethionine synthase. The expression stability of these genes was observed across the same set of tissues in ’Golden Promise’ and ‘Harrington’. These results may be broadly applicable to barley, as these cultivars are genetically diverse and commonly used.