Suitable Reference Genes/miRNAs for qRT-PCR Normalization of Expression Analysis in Sugarcane Under Sorghum mosaic virus Infection
Suitable Reference Genes/miRNAs for qRT-PCR Normalization of Expression Analysis in Sugarcane Under Sorghum mosaic virus Infection
复制标题
高粱花叶病毒感染下甘蔗表达分析 qRT-PCR 标准化的合适参考基因/miRNA
DOI：
10.1007/s12355-019-00712-1
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发表时间：
2019-10-01
期刊：
影响因子：
1.9
通讯作者：
Que, Youxiong
中科院分区：
文献类型：
--
作者：
Ling, Hui;Huang, Ning;Que, Youxiong
甘蔗花叶病对甘蔗生产构成严重威胁。许多研究旨在阐明甘蔗与花叶病毒相互作用的分子机制。定量逆转录（qRT）-PCR结合合适的内参基因已广泛用于基因表达分析。本研究首次利用geNorm、Normands和deltaCt（deltaCq）算法分析了3个甘蔗基因型感染高粱花叶病毒后叶片中33个候选参考基因和12个候选参考miRNAs的表达。将eIF-4 E和三种病毒来源的siRNA（vsiR 9230 S、vsiR 9058 A和miR 16）的表达与标准化的不稳定和稳定参考基因或miRNA进行比较，表明PP 2A和miR 159构成SrMV感染下的最佳单一参考基因/miRNA。我们还建议CUL + CAC和miR 171 + miR 1520可以作为最合适的参考基因/miRNA组合。
Sugarcane mosaic disease poses a serious threat to the sugarcane industry. Many studies have aimed to unravel the molecular mechanism related to sugarcane and mosaic virus interaction. Quantitative reverse transcription (qRT)-PCR in combination with suitable internal reference genes has been widely used for gene expression analysis. In this study, the expression of 33 candidate reference genes and 12 candidate reference miRNAs was analyzed for first time in the leaf samples of three sugarcane genotype infected with Sorghum mosaic virus using the geNorm, NormFinder and deltaCt (deltaCq) algorithms. A comparison of the expression of eIF-4E and three virus-derived siRNA (vsiR9230S, vsiR9058A and miR16) with the normalized unstable and stable reference genes or miRNA indicated that PP2A and miR159 constituted the best single reference gene/miRNA under SrMV infection. We also suggested that both CUL + CAC and miR171+ miR1520 could serve as the most suitable reference gene/miRNA combination.