Development and application of qRT-PCR for sugar beet gene expression analysis in response to in vitro induced water deficit

Sugar beet is a significant industrial crop, often grown in the areas where summer drought can severely limit root yield and sugar content. In order to improve development of sugar beet cultivars with increased drought tolerance it is necessary to understand plant response to water stress at the gen...

Descripción completa

Guardado en:

Detalles Bibliográficos
Autores principales:	Taski-Ajdukovic,Ksenija, Nagl,Nevena, Kovacev,Lazar, Curcic,Zivko, Danojevic,Dario
Lenguaje:	English
Publicado:	Pontificia Universidad Católica de Valparaíso 2012
Materias:	Beta vulgaris drought in vitro real-time PCR
Acceso en línea:	http://www.scielo.cl/scielo.php?script=sci_arttext&pid=S0717-34582012000600010
Etiquetas:	Agregar Etiqueta Sin Etiquetas, Sea el primero en etiquetar este registro!

id	oai:scielo:S0717-34582012000600010
record_format	dspace
spelling	oai:scielo:S0717-345820120006000102013-01-14Development and application of qRT-PCR for sugar beet gene expression analysis in response to in vitro induced water deficitTaski-Ajdukovic,KsenijaNagl,NevenaKovacev,LazarCurcic,ZivkoDanojevic,Dario Beta vulgaris drought in vitro real-time PCR Sugar beet is a significant industrial crop, often grown in the areas where summer drought can severely limit root yield and sugar content. In order to improve development of sugar beet cultivars with increased drought tolerance it is necessary to understand plant response to water stress at the genomic level. Since recent research efforts have focused on the molecular response of the plant in order to identify water deficit inducible genes, the aim of this investigation was to develop qRT-PCR methodology for the quantification of gene expression in sugar beet under conditions of water deficiency in vitro. Sugar beet genotypes, selected for different response to water deficit, were grown and multiplied in vitro. Axilary shoots were placed on micropropagation media with 0%, 3% and 5% PEG, for 28 days. To determine reaction of sugar beet genotypes to in vitro induced water deficit changes in number of axillary shoots, shoot fresh weight and dry matter content were measured. Total RNA was extracted from leaves and reverse transcribed into cDNA, which served as matrix in real-time PCR reaction using TaqMan technology. The housekeeping gene for glutamine synthetase was used as endogenous control, while the genes for alpha amylase and osmotin-like protein were target genes. The relative quantification values for each target gene were calculated by the 2−ΔΔCt method. Selected candidate genes differed in relative gene expression among genotypes and applied PEG treatments. The obtained results indicated that qRT-PCR protocol was efficient and accurate, showing the potential to be used in further expression analysis of candidate genes involved in sugar beet reaction to water stress.info:eu-repo/semantics/openAccessPontificia Universidad Católica de ValparaísoElectronic Journal of Biotechnology v.15 n.6 20122012-11-01text/htmlhttp://www.scielo.cl/scielo.php?script=sci_arttext&pid=S0717-34582012000600010en10.2225/vol15-issue6-fulltext-9
institution	Scielo Chile
collection	Scielo Chile
language	English
topic	Beta vulgaris drought in vitro real-time PCR
spellingShingle	Beta vulgaris drought in vitro real-time PCR Taski-Ajdukovic,Ksenija Nagl,Nevena Kovacev,Lazar Curcic,Zivko Danojevic,Dario Development and application of qRT-PCR for sugar beet gene expression analysis in response to in vitro induced water deficit
description	Sugar beet is a significant industrial crop, often grown in the areas where summer drought can severely limit root yield and sugar content. In order to improve development of sugar beet cultivars with increased drought tolerance it is necessary to understand plant response to water stress at the genomic level. Since recent research efforts have focused on the molecular response of the plant in order to identify water deficit inducible genes, the aim of this investigation was to develop qRT-PCR methodology for the quantification of gene expression in sugar beet under conditions of water deficiency in vitro. Sugar beet genotypes, selected for different response to water deficit, were grown and multiplied in vitro. Axilary shoots were placed on micropropagation media with 0%, 3% and 5% PEG, for 28 days. To determine reaction of sugar beet genotypes to in vitro induced water deficit changes in number of axillary shoots, shoot fresh weight and dry matter content were measured. Total RNA was extracted from leaves and reverse transcribed into cDNA, which served as matrix in real-time PCR reaction using TaqMan technology. The housekeeping gene for glutamine synthetase was used as endogenous control, while the genes for alpha amylase and osmotin-like protein were target genes. The relative quantification values for each target gene were calculated by the 2−ΔΔCt method. Selected candidate genes differed in relative gene expression among genotypes and applied PEG treatments. The obtained results indicated that qRT-PCR protocol was efficient and accurate, showing the potential to be used in further expression analysis of candidate genes involved in sugar beet reaction to water stress.
author	Taski-Ajdukovic,Ksenija Nagl,Nevena Kovacev,Lazar Curcic,Zivko Danojevic,Dario
author_facet	Taski-Ajdukovic,Ksenija Nagl,Nevena Kovacev,Lazar Curcic,Zivko Danojevic,Dario
author_sort	Taski-Ajdukovic,Ksenija
title	Development and application of qRT-PCR for sugar beet gene expression analysis in response to in vitro induced water deficit
title_short	Development and application of qRT-PCR for sugar beet gene expression analysis in response to in vitro induced water deficit
title_full	Development and application of qRT-PCR for sugar beet gene expression analysis in response to in vitro induced water deficit
title_fullStr	Development and application of qRT-PCR for sugar beet gene expression analysis in response to in vitro induced water deficit
title_full_unstemmed	Development and application of qRT-PCR for sugar beet gene expression analysis in response to in vitro induced water deficit
title_sort	development and application of qrt-pcr for sugar beet gene expression analysis in response to in vitro induced water deficit
publisher	Pontificia Universidad Católica de Valparaíso
publishDate	2012
url	http://www.scielo.cl/scielo.php?script=sci_arttext&pid=S0717-34582012000600010
work_keys_str_mv	AT taskiajdukovicksenija developmentandapplicationofqrtpcrforsugarbeetgeneexpressionanalysisinresponsetoinvitroinducedwaterdeficit AT naglnevena developmentandapplicationofqrtpcrforsugarbeetgeneexpressionanalysisinresponsetoinvitroinducedwaterdeficit AT kovacevlazar developmentandapplicationofqrtpcrforsugarbeetgeneexpressionanalysisinresponsetoinvitroinducedwaterdeficit AT curciczivko developmentandapplicationofqrtpcrforsugarbeetgeneexpressionanalysisinresponsetoinvitroinducedwaterdeficit AT danojevicdario developmentandapplicationofqrtpcrforsugarbeetgeneexpressionanalysisinresponsetoinvitroinducedwaterdeficit
_version_	1718441867668881408

Development and application of qRT-PCR for sugar beet gene expression analysis in response to in vitro induced water deficit

Ejemplares similares