Liu Yu, Liu Jun, Xu Lei, Lai Hui, Chen Yu, Yang Zhimin, Huang Bingru
College of Agro-grassland Science, Nanjing Agricultural University, Nanjing 210095, China.
Department of Plant Biology and Pathology, Rutgers University, New Brunswick, NJ 08901, USA.
Int J Mol Sci. 2017 Jun 21;18(6):1322. doi: 10.3390/ijms18061322.
Seashore paspalum () 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). combined with glyceraldehyde-3-phosphate dehydrogenase (GAPDH) showed stable expression in Cd-treated leaves and cold-treated roots. and 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 paired with either or were stable. and exhibited the most stability in salt-treated leaves, and combining , , and was most suitable for PEG-treated leaves. The stability of and instability of and was validated by analyzing the expression levels of four target genes (, , , and ), and were shown to be capable of detecting subtle changes in expression levels of the target genes in seashore paspalum. This study demonstrated that , , and could be used in future molecular studies that aim to understand the mechanisms of abiotic stress tolerance in seashore paspalum.
海滨雀稗（）是最耐盐和镉的暖季型多年生草本植物之一，广泛用作草坪草或饲料。本研究的目的是为海滨雀稗响应四种非生物胁迫的定量实时聚合酶链反应（qRT-PCR）分析选择稳定的内参基因。通过四个程序（geNorm、NormFinder、BestKeeper和RefFinder）评估了12个潜在内参基因的稳定性。甘油醛-3-磷酸脱氢酶（GAPDH）组合在镉处理的叶片和冷处理的根中表现出稳定的表达。和是镉处理的根和冷处理的叶片中最稳定的内参基因。在聚乙二醇（PEG）或盐处理的根中，与或配对的内参基因稳定。和在盐处理的叶片中表现出最高的稳定性，并且组合、和最适合PEG处理的叶片。通过分析四个靶基因（、、和）的表达水平验证了和的稳定性以及和的不稳定性，结果表明它们能够检测海滨雀稗中靶基因表达水平的细微变化。本研究表明，、和可用于未来旨在了解海滨雀稗非生物胁迫耐受机制的分子研究。
