Identification and validation of reference genes for quantification of target gene expression with quantitative real-time PCR for tall fescue under four abiotic stresses

PloS One
Zhimin YangBingru Huang

Abstract

Tall fescue (Festuca arundinacea Schreb.) is widely utilized as a major forage and turfgrass species in the temperate regions of the world and is a valuable plant material for studying molecular mechanisms of grass stress tolerance due to its superior drought and heat tolerance among cool-season species. Selection of suitable reference genes for quantification of target gene expression is important for the discovery of molecular mechanisms underlying improved growth traits and stress tolerance. The stability of nine potential reference genes (ACT, TUB, EF1a, GAPDH, SAND, CACS, F-box, PEPKR1 and TIP41) was evaluated using four programs, GeNorm, NormFinder, BestKeeper, and RefFinder. The combinations of SAND and TUB or TIP41 and TUB were most stably expressed in salt-treated roots or leaves. The combinations of GAPDH with TIP41 or TUB were stable in roots and leaves under drought stress. TIP41 and PEPKR1 exhibited stable expression in cold-treated roots, and the combination of F-box, TIP41 and TUB was also stable in cold-treated leaves. CACS and TUB were the two most stable reference genes in heat-stressed roots. TIP41 combined with TUB and ACT was stably expressed in heat-stressed leaves. Finally, quantitative real-time polymera...Continue Reading

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Citations

Jun 22, 2017·International Journal of Molecular Sciences·Yu LiuBingru Huang
May 1, 2019·BMC Molecular Biology·Md Sabbir HossainMd Shahidul Islam
Mar 25, 2020·Physiology and Molecular Biology of Plants : an International Journal of Functional Plant Biology·Muhammad Bilal SarwarTayyab Husnain
Aug 29, 2018·Physiology and Molecular Biology of Plants : an International Journal of Functional Plant Biology·Pradeep K JatavSumita Kachhwaha

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Datasets Mentioned

BETA
GT038376
GT051159
GT037051

Methods Mentioned

BETA
PCR
electrophoresis

Software Mentioned

Normfinder
BLAST
Primer Premier
BestKeeper
GeNorm
RefFinder
TBLASTX
LinRegPCR

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