Reliable reference genes for qPCR normalization in females of the mirid predator, Cyrtorhinus lividipennis (Hemiptera: Miridae)
Reliable reference genes for qPCR normalization in females of the mirid predator, Cyrtorhinus lividipennis (Hemiptera: Miridae)
复制标题
用于雌性食肉动物 Cyrtorhinus lividipennis（半翅目：Miridae）qPCR 标准化的可靠参考基因
DOI：
10.1007/s11756-022-01093-9
复制
发表时间：
2022-06-13
期刊：
影响因子：
1.5
通讯作者：
Liu, Fang
中科院分区：
文献类型：
--
作者：
Hu, Kui;Chen, Ke;Liu, Fang
丽纹夜蛾(Cyrtorhinus Lividipennis)是一种捕食性天敌，在调节水稻生态系统中稻飞虱和叶蝉的种群密度中起着重要作用。为了了解利氏弧菌许多生理过程的分子机制，需要对基因表达进行研究，而逆转录-定量聚合酶链式反应(RT-qPCR)是最常用的量化靶基因表达的方法。然而，目前还没有关于变铅青霉可靠参考基因的报道。在这里，我们用五种算法(Delta Ct方法、BestKeeper、GeNorm、NormFinder和RefFinder)评估了8个候选参考基因(RPL32、β-Tub、SDHA、RPL9、GAPDH、EF1α、β-Act和α-Act)在不同条件下的稳定性。结果表明，RPL32和RPL9是雌成虫各发育阶段的参考基因，RPL32和RPL9是成虫不同发育阶段的参考基因，RPL32、RPL9和β-Tub是雌虫各发育阶段的参考基因，β-Tub、GAPDH、RPL9和β-Act是5龄雌虫不同组织中可靠的参考基因，EF1α、RPL9、RPL32和β-Tub是1日龄雌成虫不同组织中稳定的参考基因，β-Tub、EF1α、RPL9、RPL32和β-Act是所有样本都适合的参考基因。这些结果突显了在不同的实验处理下识别参考基因的重要性。本研究为进一步在转录水平上研究变青虫的系统发育和生理机制奠定了基础。
The green mirid bug Cyrtorhinus lividipennis (Hemiptera: Miridae), known as a predator, plays an important role in modulating the population density of planthoppers and leafhoppers in rice ecosystem. To understand the molecular mechanisms underlying many physiological processes of C. lividipennis research on gene expression are required, and reverse transcription-quantitative polymerase chain reaction (RT-qPCR) is the most commonly used method to quantify the expression of target genes. However, there are no previous reports on reliable reference genes in C. lividipennis. Here, we evaluated the stability of eight candidate reference genes (RPL32, beta-Tub, SDHA, RPL9, GAPDH, EF1 alpha, beta-Act and alpha-Act) under different conditions with five algorithms (Delta Ct method, BestKeeper, GeNorm, NormFinder and RefFinder). Results have indicated that RPL32 and RPL9 are suitable reference genes across nymphal stages, RPL32 and RPL9 are the suggested reference genes in different developmental stages of adult females, RPL32, RPL9 and beta-Tub are suitable reference genes for all developmental stages, beta-Tub, GAPDH, RPL9 and beta-Act are reliable reference genes in different tissues of 5th instar females, EF1 alpha, RPL9, RPL32 and beta-Tub are stable reference genes in different tissues of one-day-old adult females, beta-Tub, EF1 alpha, RPL9, RPL32 and beta-Act are suitable reference genes in all samples. These results highlight the importance of identifying reference genes under different experimental treatments. Our study will facilitate further phylogenetic and physiological mechanism research at the transcriptional level in C. lividipennis.