Validation of reference genes for accurate normalization by quantitative polymerase chain reaction in sugarcane drought stress studies using two cultivars

Validation of reference genes for accurate normalization by quantitative polymerase chain reaction in sugarcane drought stress studies using two cultivars

ABSTRACT: The aim of this study was to analyze the expression of putative reference genes in sugarcane under drought stress. The varieties RB72454 and RB72910 were cultivated and the treatments control and drought stress applied to 135-day-old plants grown under field conditions. The stress level of the plants was measured by rate of photosynthesis, transpiration, and stomatal conductance. For each biological replicate, expression analyses were conducted using quantitative polymerase chain reaction for the genes α-tubulin, β-tubulin, β-actin, cyclophilin, eukaryotic elongation factor 1, glyceraldehyde 3-phosphate dehydrogenase (GAPDH), histone H3 and ubiquitin. Stability was evaluated using the geNorm, NormFinder, and BestKeeper software packages. Among the candidate genes, GAPDH was identified as the most stable in all software, indicating its suitability for gene expression studies in sugarcane undergoing drought stress; the gene β-actin was the second most stable. These findings suggest using GAPDH and β-actin for normalization in relative gene expression in sugarcane.

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Main Authors: Crystian,Dennis, Terto,Jackeline, Silva,José Vieira, Almeida,Cícero
Format: Digital revista
Language:English
Published: Universidade Federal de Santa Maria 2018
Online Access:http://old.scielo.br/scielo.php?script=sci_arttext&pid=S0103-84782018001100203
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spelling oai:scielo:S0103-847820180011002032018-11-07Validation of reference genes for accurate normalization by quantitative polymerase chain reaction in sugarcane drought stress studies using two cultivarsCrystian,DennisTerto,JackelineSilva,José VieiraAlmeida,Cícero gene expression drought stress reference gen Saccharum ABSTRACT: The aim of this study was to analyze the expression of putative reference genes in sugarcane under drought stress. The varieties RB72454 and RB72910 were cultivated and the treatments control and drought stress applied to 135-day-old plants grown under field conditions. The stress level of the plants was measured by rate of photosynthesis, transpiration, and stomatal conductance. For each biological replicate, expression analyses were conducted using quantitative polymerase chain reaction for the genes α-tubulin, β-tubulin, β-actin, cyclophilin, eukaryotic elongation factor 1, glyceraldehyde 3-phosphate dehydrogenase (GAPDH), histone H3 and ubiquitin. Stability was evaluated using the geNorm, NormFinder, and BestKeeper software packages. Among the candidate genes, GAPDH was identified as the most stable in all software, indicating its suitability for gene expression studies in sugarcane undergoing drought stress; the gene β-actin was the second most stable. These findings suggest using GAPDH and β-actin for normalization in relative gene expression in sugarcane.info:eu-repo/semantics/openAccessUniversidade Federal de Santa MariaCiência Rural v.48 n.11 20182018-01-01info:eu-repo/semantics/articletext/htmlhttp://old.scielo.br/scielo.php?script=sci_arttext&pid=S0103-84782018001100203en10.1590/0103-8478cr20180152
institution SCIELO
collection OJS
country Brasil
countrycode BR
component Revista
access En linea
databasecode rev-scielo-br
tag revista
region America del Sur
libraryname SciELO
language English
format Digital
author Crystian,Dennis
Terto,Jackeline
Silva,José Vieira
Almeida,Cícero
spellingShingle Crystian,Dennis
Terto,Jackeline
Silva,José Vieira
Almeida,Cícero
Validation of reference genes for accurate normalization by quantitative polymerase chain reaction in sugarcane drought stress studies using two cultivars
author_facet Crystian,Dennis
Terto,Jackeline
Silva,José Vieira
Almeida,Cícero
author_sort Crystian,Dennis
title Validation of reference genes for accurate normalization by quantitative polymerase chain reaction in sugarcane drought stress studies using two cultivars
title_short Validation of reference genes for accurate normalization by quantitative polymerase chain reaction in sugarcane drought stress studies using two cultivars
title_full Validation of reference genes for accurate normalization by quantitative polymerase chain reaction in sugarcane drought stress studies using two cultivars
title_fullStr Validation of reference genes for accurate normalization by quantitative polymerase chain reaction in sugarcane drought stress studies using two cultivars
title_full_unstemmed Validation of reference genes for accurate normalization by quantitative polymerase chain reaction in sugarcane drought stress studies using two cultivars
title_sort validation of reference genes for accurate normalization by quantitative polymerase chain reaction in sugarcane drought stress studies using two cultivars
description ABSTRACT: The aim of this study was to analyze the expression of putative reference genes in sugarcane under drought stress. The varieties RB72454 and RB72910 were cultivated and the treatments control and drought stress applied to 135-day-old plants grown under field conditions. The stress level of the plants was measured by rate of photosynthesis, transpiration, and stomatal conductance. For each biological replicate, expression analyses were conducted using quantitative polymerase chain reaction for the genes α-tubulin, β-tubulin, β-actin, cyclophilin, eukaryotic elongation factor 1, glyceraldehyde 3-phosphate dehydrogenase (GAPDH), histone H3 and ubiquitin. Stability was evaluated using the geNorm, NormFinder, and BestKeeper software packages. Among the candidate genes, GAPDH was identified as the most stable in all software, indicating its suitability for gene expression studies in sugarcane undergoing drought stress; the gene β-actin was the second most stable. These findings suggest using GAPDH and β-actin for normalization in relative gene expression in sugarcane.
publisher Universidade Federal de Santa Maria
publishDate 2018
url http://old.scielo.br/scielo.php?script=sci_arttext&pid=S0103-84782018001100203
work_keys_str_mv AT crystiandennis validationofreferencegenesforaccuratenormalizationbyquantitativepolymerasechainreactioninsugarcanedroughtstressstudiesusingtwocultivars
AT tertojackeline validationofreferencegenesforaccuratenormalizationbyquantitativepolymerasechainreactioninsugarcanedroughtstressstudiesusingtwocultivars
AT silvajosevieira validationofreferencegenesforaccuratenormalizationbyquantitativepolymerasechainreactioninsugarcanedroughtstressstudiesusingtwocultivars
AT almeidacicero validationofreferencegenesforaccuratenormalizationbyquantitativepolymerasechainreactioninsugarcanedroughtstressstudiesusingtwocultivars
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