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Biblioteca (s) : |
INIA Las Brujas. |
Fecha : |
11/03/2021 |
Actualizado : |
11/03/2021 |
Tipo de producción científica : |
Artículos en Revistas Indexadas Internacionales |
Autor : |
NARANCIO, R.; JOHN, U.; MASON, J.; SPANGENBERG, G. |
Afiliación : |
RAFAEL NARANCIO FERES, Agriculture Victoria Research, AgriBio Centre for AgriBioscience, La Trobe University, Melbourne, Australia; School of Applied Systems Biology, La Trobe University, Melbourne, Australia; ULRIK JOHN, Agriculture Victoria Research, AgriBio Centre for AgriBioscience, La Trobe University, Melbourne, Australia; JOHN MASON, Agriculture Victoria Research, AgriBio Centre for AgriBioscience, La Trobe University, Melbourne, Australia; School of Applied Systems Biology, La Trobe University, Melbourne, Australia; GERMAN SPANGENBERG, Agriculture Victoria Research, AgriBio Centre for AgriBioscience, La Trobe University, Melbourne, Australia; School of Applied Systems Biology, La Trobe University, Melbourne, Australia. |
Título : |
Selection of optimal reference genes for quantitative RT-PCR transcript abundance analysis in white clover (Trifolium repens L.). |
Fecha de publicación : |
2018 |
Fuente / Imprenta : |
Functional Plant Biology, 2018, Volume 45, Issue 7, Pages 737-744. Doi: https://doi.org/10.1071/FP17304 |
ISSN : |
1445-4408 |
DOI : |
10.1071/FP17304 |
Idioma : |
Inglés |
Notas : |
Article history: Received 1 November 2017; Accepted 21 January 2018; Published online 16 February 2018.
Corresponding author: John, U.; Agriculture Victoria Research, AgriBio Centre for AgriBioscience, La Trobe University, Melbourne, VIC, Australia; email:ulrik.john@ecodev.vic.gov.au |
Contenido : |
ABSTRACT -
Quantitative reverse transcription PCR (qRT-PCR) is a widely used method for transcript abundance analyses in plants. Relative quantification by qRT-PCR requires the use of a stably expressed reference gene. There are many ?housekeeping? genes reported in different plant species that are used as reference genes. However, it is important that the steady-state mRNA levels of these housekeeping genes are confirmed across different conditions and tissues in each species studied. Prior to this study, no comprehensive work had been performed in identifying optimal reference genes in white clover (Trifolium repens L.). To identify suitable reference genes in white clover, we analysed the transcript abundance stability of seven candidate genes in two organs (leaves and stolons) across two treatments (water-limited and well-watered). DCt, NormFinder and ANOVA tests were carried out to evaluate the mRNA level stability of candidate reference genes. According to the DCt results, the genes with the most stable mRNA levels were EF1a and ACT11. When stability among groups was evaluated by NormFinder, UBQ was the most stable across all organs and treatments.
By multiple criteria, EF1a, followed by ACT11 and UBQ, was the most stably-expressed gene across organs and treatments, and each of these are recommended as reference genes for transcript abundance studies in white clover.
© Copyright 2018 Elsevier B.V., All rights reserved. |
Palabras claves : |
ACt; Gene Selection; Housekeeping genes; MRNA level stability; NormFinder; Peptide Elongation Factor 1. |
Asunto categoría : |
F01 Cultivo |
Marc : |
LEADER 02570naa a2200265 a 4500 001 1061825 005 2021-03-11 008 2018 bl uuuu u00u1 u #d 022 $a1445-4408 024 7 $a10.1071/FP17304$2DOI 100 1 $aNARANCIO, R. 245 $aSelection of optimal reference genes for quantitative RT-PCR transcript abundance analysis in white clover (Trifolium repens L.).$h[electronic resource] 260 $c2018 500 $aArticle history: Received 1 November 2017; Accepted 21 January 2018; Published online 16 February 2018. Corresponding author: John, U.; Agriculture Victoria Research, AgriBio Centre for AgriBioscience, La Trobe University, Melbourne, VIC, Australia; email:ulrik.john@ecodev.vic.gov.au 520 $aABSTRACT - Quantitative reverse transcription PCR (qRT-PCR) is a widely used method for transcript abundance analyses in plants. Relative quantification by qRT-PCR requires the use of a stably expressed reference gene. There are many ?housekeeping? genes reported in different plant species that are used as reference genes. However, it is important that the steady-state mRNA levels of these housekeeping genes are confirmed across different conditions and tissues in each species studied. Prior to this study, no comprehensive work had been performed in identifying optimal reference genes in white clover (Trifolium repens L.). To identify suitable reference genes in white clover, we analysed the transcript abundance stability of seven candidate genes in two organs (leaves and stolons) across two treatments (water-limited and well-watered). DCt, NormFinder and ANOVA tests were carried out to evaluate the mRNA level stability of candidate reference genes. According to the DCt results, the genes with the most stable mRNA levels were EF1a and ACT11. When stability among groups was evaluated by NormFinder, UBQ was the most stable across all organs and treatments. By multiple criteria, EF1a, followed by ACT11 and UBQ, was the most stably-expressed gene across organs and treatments, and each of these are recommended as reference genes for transcript abundance studies in white clover. © Copyright 2018 Elsevier B.V., All rights reserved. 653 $aACt 653 $aGene Selection 653 $aHousekeeping genes 653 $aMRNA level stability 653 $aNormFinder 653 $aPeptide Elongation Factor 1 700 1 $aJOHN, U. 700 1 $aMASON, J. 700 1 $aSPANGENBERG, G. 773 $tFunctional Plant Biology, 2018, Volume 45, Issue 7, Pages 737-744. Doi: https://doi.org/10.1071/FP17304
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