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Int. J. Mol. Sci. 2017, 18(6), 1322; doi:10.3390/ijms18061322

Identification and Validation of Reference Genes for Seashore Paspalum Response to Abiotic Stresses

1
College of Agro-grassland Science, Nanjing Agricultural University, Nanjing 210095, China
2
Department of Plant Biology and Pathology, Rutgers University, New Brunswick, NJ 08901, USA
*
Authors to whom correspondence should be addressed.
Received: 19 April 2017 / Revised: 16 June 2017 / Accepted: 16 June 2017 / Published: 21 June 2017
(This article belongs to the Section Molecular Botany)
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Abstract

Seashore paspalum (Paspalum vaginatum) is among the most salt- and cadmium-tolerant warm-season perennial grass species widely used as turf or forage. The objective of this study was to select stable reference genes for quantitative real-time polymerase chain reaction (qRT-PCR) analysis of seashore paspalum in response to four abiotic stresses. The stability of 12 potential reference genes was evaluated by four programs (geNorm, NormFinder, BestKeeper, and RefFinder). U2AF combined with glyceraldehyde-3-phosphate dehydrogenase (GAPDH) showed stable expression in Cd-treated leaves and cold-treated roots. U2AF and FBOX were the most stable reference genes in Cd-treated roots and cold-treated leaves. In Polyethylene Glycol (PEG)- or salt-treated roots, the reference gene U2AF paired with either ACT or CYP were stable. SAND and CACS exhibited the most stability in salt-treated leaves, and combining UPL, PP2A, and EF1a was most suitable for PEG-treated leaves. The stability of U2AF and instability of UPL and TUB was validated by analyzing the expression levels of four target genes (MT2a, VP1, PIP1, and Cor413), and were shown to be capable of detecting subtle changes in expression levels of the target genes in seashore paspalum. This study demonstrated that FBOX, U2AF, and PP2A could be used in future molecular studies that aim to understand the mechanisms of abiotic stress tolerance in seashore paspalum. View Full-Text
Keywords: seashore paspalum; quantitative real-time polymerase chain reaction (qRT-PCR); reference gene; abiotic stress seashore paspalum; quantitative real-time polymerase chain reaction (qRT-PCR); reference gene; abiotic stress
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MDPI and ACS Style

Liu, Y.; Liu, J.; Xu, L.; Lai, H.; Chen, Y.; Yang, Z.; Huang, B. Identification and Validation of Reference Genes for Seashore Paspalum Response to Abiotic Stresses. Int. J. Mol. Sci. 2017, 18, 1322.

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