渗透压胁迫：通过 RT-qPCR 进行转录谱分析的参考基因鉴定。

and Osmotic Stress: Identification of Reference Genes for Transcriptional Profiling by RT-qPCR.

机构信息

Institute of Plant Genetics, Breeding and Biotechnology, University of Life Sciences in Lublin, 20-612 Lublin, Poland.

出版信息

Genes (Basel). 2020 Aug 31;11(9):1022. doi: 10.3390/genes11091022.

DOI:10.3390/genes11091022

PMID:32878033

原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC7563777/

Abstract

is a highly valued perennial ornamental grass cultivated worldwide. However, there is limited genetic data regarding this species. Selection of proper reference genes (RGs) for reverse transcription quantitative PCR (RT-qPCR) data normalization has become an essential step in gene expression analysis. In this study, we aimed to examine expression stability of nine candidate RGs in plants, subjected to osmotic stress, generated either by salinity or PEG treatment. The identification of genes exhibiting high expression stability was performed by four algorithms (geNorm, NormFinder, BestKeeper and deltaCt method). The results showed that the combination of two genes would be sufficient for reliable expression data normalization. ADP (ADP-ribosylation factor) and TBP (TATA-box-binding protein) were identified as the most stably expressed under salinity treatment, while eIF4A (eukaryotic initiation factor 4A) and were found to show the highest stability under PEG-induced drought. A set of three genes (, and ) displayed the highest expression stability across all experimental samples tested in this study. To our best knowledge, this is the first report regarding RGs selection in . It will provide valuable starting point information for conducting further analyses in this and related species concerning their responses to water shortage and salinity stress.

摘要

是一种备受推崇的多年生观赏草，在全球范围内被广泛种植。然而，关于该物种的遗传数据有限。选择合适的内参基因（RGs）进行逆转录定量 PCR（RT-qPCR）数据归一化已成为基因表达分析的重要步骤。在本研究中，我们旨在研究 9 种候选 RGs 在盐胁迫或 PEG 处理下的表达稳定性。通过四种算法（geNorm、NormFinder、BestKeeper 和 deltaCt 方法）确定具有高表达稳定性的基因。结果表明，使用两个基因的组合即可进行可靠的表达数据归一化。在盐胁迫处理下，ADP（ADP-ribosylation factor）和 TBP（TATA 盒结合蛋白）被鉴定为最稳定表达的基因，而 eIF4A（真核起始因子 4A）和则在 PEG 诱导的干旱条件下表现出最高的稳定性。一组三个基因（、和）在本研究中测试的所有实验样本中表现出最高的表达稳定性。据我们所知，这是首次在中报告 RGs 选择。它将为进一步研究该物种及相关物种对缺水和盐胁迫的响应提供有价值的起点信息。