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Genes 2016, 7(6), 25; doi:10.3390/genes7060025

Selection and Verification of Candidate Reference Genes for Mature MicroRNA Expression by Quantitative RT-PCR in the Tea Plant (Camellia sinensis)

State Key Laboratory of Tea Plant Biology and Utilization, Anhui Agricultural University, 130 Changjiang West Road, Hefei 230036, China
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Author to whom correspondence should be addressed.
Academic Editors: Lukas Mueller and Peter W. Young
Received: 27 February 2016 / Revised: 11 April 2016 / Accepted: 24 May 2016 / Published: 28 May 2016
(This article belongs to the Section Population and Evolutionary Genetics and Genomics)
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Abstract

Quantitative reverse transcription-polymerase chain reaction (qRT-PCR) is a rapid and sensitive method for analyzing microRNA (miRNA) expression. However, accurate qRT-PCR results depend on the selection of reliable reference genes as internal positive controls. To date, few studies have identified reliable reference genes for differential expression analysis of miRNAs among tissues, and among experimental conditions in plants. In this study, three miRNAs and four non-coding small RNAs (ncRNA) were selected as reference candidates, and the stability of their expression was evaluated among different tissues and under different experimental conditions in the tea plant (Camellia sinensis) using the geNorm and NormFinder programs. It was shown that miR159a was the best single reference gene in the bud to the fifth leaf, 5S rRNA was the most suitable gene in different organs, miR6149 was the most stable gene when the leaves were attacked by Ectropis oblique and U4, miR5368n and miR159a were the best genes when the leaves were treated by methyl jasmonate (MeJA), salicylic acid (SA) and abscisic acid (ABA), respectively. Our results provide suitable reference genes for future investigations on miRNA functions in tea plants. View Full-Text
Keywords: Camellia sinensis; microRNA; candidate reference gene; qRT-PCR; tea plant; expression Camellia sinensis; microRNA; candidate reference gene; qRT-PCR; tea plant; expression
This is an open access article distributed under the Creative Commons Attribution License which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. (CC BY 4.0).

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MDPI and ACS Style

Song, H.; Zhang, X.; Shi, C.; Wang, S.; Wu, A.; Wei, C. Selection and Verification of Candidate Reference Genes for Mature MicroRNA Expression by Quantitative RT-PCR in the Tea Plant (Camellia sinensis). Genes 2016, 7, 25.

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