Development and application of qRT-PCR for sugar beet gene expression analysis in response to in vitro induced water deficit
Апстракт
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(-Delta Delta Ct) 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.
Кључне речи:
Beta vulgaris / drought / in vitro / real-time PCRИзвор:
Electronic Journal of Biotechnology, 2012, 15, 6Издавач:
- Univ Catolica De Valparaiso, Valparaiso
Финансирање / пројекти:
- Побољшање линија, хибрида и технологије гајења шећерне репе (RS-MESTD-Technological Development (TD or TR)-31015)
DOI: 10.2225/vol15-issue6-fulltext-9
ISSN: 0717-3458
WoS: 000314276100002
Scopus: 2-s2.0-84870026485
Колекције
Институција/група
FiVeRTY - JOUR AU - Taški-Ajduković, Ksenija AU - Nagl, Nevena AU - Kovačev, Lazar AU - Ćurčić, Živko AU - Danojević, Dario PY - 2012 UR - http://fiver.ifvcns.rs/handle/123456789/1083 AB - 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(-Delta Delta Ct) 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. PB - Univ Catolica De Valparaiso, Valparaiso T2 - Electronic Journal of Biotechnology T1 - Development and application of qRT-PCR for sugar beet gene expression analysis in response to in vitro induced water deficit IS - 6 VL - 15 DO - 10.2225/vol15-issue6-fulltext-9 ER -
@article{ author = "Taški-Ajduković, Ksenija and Nagl, Nevena and Kovačev, Lazar and Ćurčić, Živko and Danojević, Dario", year = "2012", abstract = "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(-Delta Delta Ct) 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.", publisher = "Univ Catolica De Valparaiso, Valparaiso", journal = "Electronic Journal of Biotechnology", title = "Development and application of qRT-PCR for sugar beet gene expression analysis in response to in vitro induced water deficit", number = "6", volume = "15", doi = "10.2225/vol15-issue6-fulltext-9" }
Taški-Ajduković, K., Nagl, N., Kovačev, L., Ćurčić, Ž.,& Danojević, D.. (2012). Development and application of qRT-PCR for sugar beet gene expression analysis in response to in vitro induced water deficit. in Electronic Journal of Biotechnology Univ Catolica De Valparaiso, Valparaiso., 15(6). https://doi.org/10.2225/vol15-issue6-fulltext-9
Taški-Ajduković K, Nagl N, Kovačev L, Ćurčić Ž, Danojević D. Development and application of qRT-PCR for sugar beet gene expression analysis in response to in vitro induced water deficit. in Electronic Journal of Biotechnology. 2012;15(6). doi:10.2225/vol15-issue6-fulltext-9 .
Taški-Ajduković, Ksenija, Nagl, Nevena, Kovačev, Lazar, Ćurčić, Živko, Danojević, Dario, "Development and application of qRT-PCR for sugar beet gene expression analysis in response to in vitro induced water deficit" in Electronic Journal of Biotechnology, 15, no. 6 (2012), https://doi.org/10.2225/vol15-issue6-fulltext-9 . .