Reference Gene Selection for qPCR Analysis in Tomato-Bipartite Begomovirus Interaction and Validation in Additional Tomato-Virus Pathosystems

PloS One
Ana L M LacerdaAna C M Brasileiro

Abstract

Quantitative Polymerase Chain Reaction (qPCR) is currently the most sensitive technique used for absolute and relative quantification of a target gene transcript, requiring the use of appropriated reference genes for data normalization. To accurately estimate the relative expression of target tomato (Solanum lycopersicum L.) genes responsive to several virus species in reverse transcription qPCR analysis, the identification of reliable reference genes is mandatory. In the present study, ten reference genes were analyzed across a set of eight samples: two tomato contrasting genotypes ('Santa Clara', susceptible, and its near-isogenic line 'LAM 157', resistant); subjected to two treatments (inoculation with Tomato chlorotic mottle virus (ToCMoV) and its mock-inoculated control) and in two distinct times after inoculation (early and late). Reference genes stability was estimated by three statistical programs (geNorm, NormFinder and BestKeeper). To validate the results over broader experimental conditions, a set of ten samples, corresponding to additional three tomato-virus pathosystems that included tospovirus, crinivirus and tymovirus + tobamovirus, was analyzed together with the tomato-ToCMoV pathosystem dataset, using the same ...Continue Reading

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Citations

Sep 30, 2016·Frontiers in Plant Science·Karla L González-AguileraStefan de Folter
Dec 25, 2017·Archives of Virology·Mozhgan HortamaniKeramat Izadpanah
Mar 8, 2018·International Journal of Molecular Medicine·Sheng WangXiongwen Lv
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May 11, 2021·Molecular Plant-microbe Interactions : MPMI·Hagit Hak, Ziv Spiegelman

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Methods Mentioned

BETA
Polymerase Chain Reaction
PCR

Software Mentioned

real
geNorm
NormFinder
PCR Miner
geNorm PLUS
REST
BestKeeper
Primer3Plus

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