Abstract:
KEY MESSAGE:MicroRNAs have higher expression stability than protein-coding genes in B. napus seeds and are therefore good reference genes for miRNA and mRNA RT-qPCR analysis. Reverse transcription quantitative real-time polymerase chain reaction (RT-qPCR) has become the "gold standard" to gain insight into function of genes. However, the accuracy of the technique depends on appropriate reference genes for quantification analysis in different experimental conditions. Accumulation of microRNAs (miRNAs) has also been studied by RT-qPCR, but there are no reference genes currently validated for normalization of Brassica napus miRNA expression data. In this study, we selected 43 B. napus miRNAs and 18 previously validated mRNA reference genes. The expression stability of the candidate reference genes was evaluated in different tissue samples (stages of seed development, flowers, and leaves) using geNorm, NormFinder, and RefFinder analysis. The best-ranked reference genes for expression studies during seed development (miR167-1_2, miR11-1, miR159-1 and miR168-1) were used to asses the expression of miR03-1. Since candidate miRNAs showed higher expression stability than protein-coding genes in most of the tested conditions, the expression profile of DGAT1 gene was compared when normalized by the four most stable miRNAs reference genes and by the four most stable mRNA reference genes. The expected expression pattern of DGAT1 during seed development was achieved with the use of miRNA as reference genes. In conclusion, the most stable miRNA reference genes can be employed in the normalization of RT-qPCR quantification of miRNAs and protein-coding genes. This work is the first to perform a comprehensive survey of the stability of miRNA reference genes in B. napus and provides guidelines to obtain more accurate RT-qPCR results in B. napus seeds studies.
journal_name
Plant Cell Repjournal_title
Plant cell reportsauthors
Machado RD,Christoff AP,Loss-Morais G,Margis-Pinheiro M,Margis R,Körbes APdoi
10.1007/s00299-015-1773-1subject
Has Abstractpub_date
2015-07-01 00:00:00pages
1139-49issue
7eissn
0721-7714issn
1432-203Xjournal_volume
34pub_type
杂志文章abstract::High-frequency bud break and multiple shoots were induced in apical shoot buds and nodal explants ofMorus cathayana, M. lhou andM. serrata on Murashige and Skoog (MS) medium containing 0.5-1.0 mg/l 6-benzylaminopurine (BAP). Addition of gibberellic acid (0.4 mg/l) along with BAP induced faster bud break both in apical...
journal_title:Plant cell reports
pub_type: 杂志文章
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journal_title:Plant cell reports
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journal_title:Plant cell reports
pub_type: 杂志文章
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journal_title:Plant cell reports
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journal_title:Plant cell reports
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journal_title:Plant cell reports
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journal_title:Plant cell reports
pub_type: 杂志文章
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journal_title:Plant cell reports
pub_type: 杂志文章
doi:10.1007/BF00232295
更新日期:1990-09-01 00:00:00
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journal_title:Plant cell reports
pub_type: 杂志文章
doi:10.1007/s00299-004-0911-y
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journal_title:Plant cell reports
pub_type: 杂志文章
doi:10.1007/BF00716827
更新日期:1989-03-01 00:00:00
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pub_type: 杂志文章
doi:10.1007/s00299-016-1984-0
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journal_title:Plant cell reports
pub_type: 杂志文章
doi:10.1007/s00299-004-0879-7
更新日期:2005-02-01 00:00:00
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journal_title:Plant cell reports
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doi:10.1007/s00299-012-1355-4
更新日期:2013-02-01 00:00:00
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pub_type: 杂志文章
doi:10.1007/s00299-010-0909-6
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journal_title:Plant cell reports
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journal_title:Plant cell reports
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journal_title:Plant cell reports
pub_type: 杂志文章
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journal_title:Plant cell reports
pub_type: 杂志文章
doi:10.1007/BF00778536
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journal_title:Plant cell reports
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journal_title:Plant cell reports
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journal_title:Plant cell reports
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更新日期:2015-10-01 00:00:00
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journal_title:Plant cell reports
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journal_title:Plant cell reports
pub_type: 杂志文章,评审
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更新日期:2014-05-01 00:00:00
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journal_title:Plant cell reports
pub_type: 杂志文章
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更新日期:2016-11-01 00:00:00
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更新日期:1990-12-01 00:00:00