Selection of Reference Genes for RT-qPCR Studies in Different Organs of Rice Cultivar BRS AG Submitted to Recurrent Saline Stress.
Autor(a) principal: | |
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Data de Publicação: | 2021 |
Outros Autores: | , , , , |
Tipo de documento: | Artigo |
Idioma: | eng |
Título da fonte: | Repositório Institucional da EMBRAPA (Repository Open Access to Scientific Information from EMBRAPA - Alice) |
Texto Completo: | http://www.alice.cnptia.embrapa.br/alice/handle/doc/1130778 |
Resumo: | Quantitative real-time polymerase chain reactions (RT-qPCR) have become one of the most widely used methods for analyzing gene expression, provided suitable reference genes are available to normalize the data. RNA was isolated from leaves, grain, rachises and sheaths of rice (Oryza sativa L. cv. BRS AG) submitted to different saline stress events for seven days, and expression analysis was carried out by RT-qPCR. Expression levels of ten candidate reference genes were assessed, actin11 (ACT11), ubiquitin conjugating enzyme E2 (UBC-E2), eukaryotic elongation factor1-a (Eef-1a), glyceraldehyde-3-phosphate dehydrogenase (GAPDH), B-tubulin (B-Tub), eukaryotic initiation factor 4a (Eif-4-a), ubiquitin10 (UBQ10), ubiquitin5 (UBQ5), aquaporin TIP41 (TIP41-like). Gene expression stability was calculated using the common statistical algorithms geNorm, BestKeeper and ?Ct method, NormFinder and RefFinder. The most stably expressed genes were UBC2E and GAPDH for leaves, UBQ5 and UBQ10 for sheaths, TIP41 and UBQ10 for rachises, and TIP41 and cyclophilin for grain. Gene expression of triose phosphate translocator (TPT1), ADP-glucose transporter (BT1-1), choline monooxygenase (CMO) was used to validate the selected reference genes. The results highlighted the importance of using suitable reference gene to normalize gene expression data in rice plants. |
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Selection of Reference Genes for RT-qPCR Studies in Different Organs of Rice Cultivar BRS AG Submitted to Recurrent Saline Stress.ArrozGeneOryza SativaQuantitative real-time polymerase chain reactions (RT-qPCR) have become one of the most widely used methods for analyzing gene expression, provided suitable reference genes are available to normalize the data. RNA was isolated from leaves, grain, rachises and sheaths of rice (Oryza sativa L. cv. BRS AG) submitted to different saline stress events for seven days, and expression analysis was carried out by RT-qPCR. Expression levels of ten candidate reference genes were assessed, actin11 (ACT11), ubiquitin conjugating enzyme E2 (UBC-E2), eukaryotic elongation factor1-a (Eef-1a), glyceraldehyde-3-phosphate dehydrogenase (GAPDH), B-tubulin (B-Tub), eukaryotic initiation factor 4a (Eif-4-a), ubiquitin10 (UBQ10), ubiquitin5 (UBQ5), aquaporin TIP41 (TIP41-like). Gene expression stability was calculated using the common statistical algorithms geNorm, BestKeeper and ?Ct method, NormFinder and RefFinder. The most stably expressed genes were UBC2E and GAPDH for leaves, UBQ5 and UBQ10 for sheaths, TIP41 and UBQ10 for rachises, and TIP41 and cyclophilin for grain. Gene expression of triose phosphate translocator (TPT1), ADP-glucose transporter (BT1-1), choline monooxygenase (CMO) was used to validate the selected reference genes. The results highlighted the importance of using suitable reference gene to normalize gene expression data in rice plants.T. ROSSATTO, UFPEL; P. A. AULER, UFPEL; M. N. AMARAL, UFPEL; C. MILECH, UFPEL; ARIANO MARTINS DE MAGALHAES JUNIOR, CPACT; E. J. B. BRAGA, UFPEL.ROSSATTO, T.AULER, P. A.AMARAL, M. N.MILECH, C.MAGALHAES JUNIOR, A. M. deBRAGA, E. J. B.2021-03-18T14:36:42Z2021-03-18T14:36:42Z2021-03-182021info:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/articleRussian Journal of Plant Physiology, v. 68, n. 2, p. 254-265, 2021.1021-4437http://www.alice.cnptia.embrapa.br/alice/handle/doc/1130778enginfo:eu-repo/semantics/openAccessreponame:Repositório Institucional da EMBRAPA (Repository Open Access to Scientific Information from EMBRAPA - Alice)instname:Empresa Brasileira de Pesquisa Agropecuária (Embrapa)instacron:EMBRAPA2021-03-18T14:36:52Zoai:www.alice.cnptia.embrapa.br:doc/1130778Repositório InstitucionalPUBhttps://www.alice.cnptia.embrapa.br/oai/requestopendoar:21542021-03-18T14:36:52falseRepositório InstitucionalPUBhttps://www.alice.cnptia.embrapa.br/oai/requestcg-riaa@embrapa.bropendoar:21542021-03-18T14:36:52Repositório Institucional da EMBRAPA (Repository Open Access to Scientific Information from EMBRAPA - Alice) - Empresa Brasileira de Pesquisa Agropecuária (Embrapa)false |
dc.title.none.fl_str_mv |
Selection of Reference Genes for RT-qPCR Studies in Different Organs of Rice Cultivar BRS AG Submitted to Recurrent Saline Stress. |
title |
Selection of Reference Genes for RT-qPCR Studies in Different Organs of Rice Cultivar BRS AG Submitted to Recurrent Saline Stress. |
spellingShingle |
Selection of Reference Genes for RT-qPCR Studies in Different Organs of Rice Cultivar BRS AG Submitted to Recurrent Saline Stress. ROSSATTO, T. Arroz Gene Oryza Sativa |
title_short |
Selection of Reference Genes for RT-qPCR Studies in Different Organs of Rice Cultivar BRS AG Submitted to Recurrent Saline Stress. |
title_full |
Selection of Reference Genes for RT-qPCR Studies in Different Organs of Rice Cultivar BRS AG Submitted to Recurrent Saline Stress. |
title_fullStr |
Selection of Reference Genes for RT-qPCR Studies in Different Organs of Rice Cultivar BRS AG Submitted to Recurrent Saline Stress. |
title_full_unstemmed |
Selection of Reference Genes for RT-qPCR Studies in Different Organs of Rice Cultivar BRS AG Submitted to Recurrent Saline Stress. |
title_sort |
Selection of Reference Genes for RT-qPCR Studies in Different Organs of Rice Cultivar BRS AG Submitted to Recurrent Saline Stress. |
author |
ROSSATTO, T. |
author_facet |
ROSSATTO, T. AULER, P. A. AMARAL, M. N. MILECH, C. MAGALHAES JUNIOR, A. M. de BRAGA, E. J. B. |
author_role |
author |
author2 |
AULER, P. A. AMARAL, M. N. MILECH, C. MAGALHAES JUNIOR, A. M. de BRAGA, E. J. B. |
author2_role |
author author author author author |
dc.contributor.none.fl_str_mv |
T. ROSSATTO, UFPEL; P. A. AULER, UFPEL; M. N. AMARAL, UFPEL; C. MILECH, UFPEL; ARIANO MARTINS DE MAGALHAES JUNIOR, CPACT; E. J. B. BRAGA, UFPEL. |
dc.contributor.author.fl_str_mv |
ROSSATTO, T. AULER, P. A. AMARAL, M. N. MILECH, C. MAGALHAES JUNIOR, A. M. de BRAGA, E. J. B. |
dc.subject.por.fl_str_mv |
Arroz Gene Oryza Sativa |
topic |
Arroz Gene Oryza Sativa |
description |
Quantitative real-time polymerase chain reactions (RT-qPCR) have become one of the most widely used methods for analyzing gene expression, provided suitable reference genes are available to normalize the data. RNA was isolated from leaves, grain, rachises and sheaths of rice (Oryza sativa L. cv. BRS AG) submitted to different saline stress events for seven days, and expression analysis was carried out by RT-qPCR. Expression levels of ten candidate reference genes were assessed, actin11 (ACT11), ubiquitin conjugating enzyme E2 (UBC-E2), eukaryotic elongation factor1-a (Eef-1a), glyceraldehyde-3-phosphate dehydrogenase (GAPDH), B-tubulin (B-Tub), eukaryotic initiation factor 4a (Eif-4-a), ubiquitin10 (UBQ10), ubiquitin5 (UBQ5), aquaporin TIP41 (TIP41-like). Gene expression stability was calculated using the common statistical algorithms geNorm, BestKeeper and ?Ct method, NormFinder and RefFinder. The most stably expressed genes were UBC2E and GAPDH for leaves, UBQ5 and UBQ10 for sheaths, TIP41 and UBQ10 for rachises, and TIP41 and cyclophilin for grain. Gene expression of triose phosphate translocator (TPT1), ADP-glucose transporter (BT1-1), choline monooxygenase (CMO) was used to validate the selected reference genes. The results highlighted the importance of using suitable reference gene to normalize gene expression data in rice plants. |
publishDate |
2021 |
dc.date.none.fl_str_mv |
2021-03-18T14:36:42Z 2021-03-18T14:36:42Z 2021-03-18 2021 |
dc.type.driver.fl_str_mv |
info:eu-repo/semantics/publishedVersion info:eu-repo/semantics/article |
format |
article |
status_str |
publishedVersion |
dc.identifier.uri.fl_str_mv |
Russian Journal of Plant Physiology, v. 68, n. 2, p. 254-265, 2021. 1021-4437 http://www.alice.cnptia.embrapa.br/alice/handle/doc/1130778 |
identifier_str_mv |
Russian Journal of Plant Physiology, v. 68, n. 2, p. 254-265, 2021. 1021-4437 |
url |
http://www.alice.cnptia.embrapa.br/alice/handle/doc/1130778 |
dc.language.iso.fl_str_mv |
eng |
language |
eng |
dc.rights.driver.fl_str_mv |
info:eu-repo/semantics/openAccess |
eu_rights_str_mv |
openAccess |
dc.source.none.fl_str_mv |
reponame:Repositório Institucional da EMBRAPA (Repository Open Access to Scientific Information from EMBRAPA - Alice) instname:Empresa Brasileira de Pesquisa Agropecuária (Embrapa) instacron:EMBRAPA |
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Empresa Brasileira de Pesquisa Agropecuária (Embrapa) |
instacron_str |
EMBRAPA |
institution |
EMBRAPA |
reponame_str |
Repositório Institucional da EMBRAPA (Repository Open Access to Scientific Information from EMBRAPA - Alice) |
collection |
Repositório Institucional da EMBRAPA (Repository Open Access to Scientific Information from EMBRAPA - Alice) |
repository.name.fl_str_mv |
Repositório Institucional da EMBRAPA (Repository Open Access to Scientific Information from EMBRAPA - Alice) - Empresa Brasileira de Pesquisa Agropecuária (Embrapa) |
repository.mail.fl_str_mv |
cg-riaa@embrapa.br |
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1794503503833464832 |