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首页> 外文期刊>Journal of Economic Entomology >Reference Gene Validation for Quantitative PCR Under Various Biotic and Abiotic Stress Conditions in Toxoptera citricida (Hemiptera, Aphidiae)
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Reference Gene Validation for Quantitative PCR Under Various Biotic and Abiotic Stress Conditions in Toxoptera citricida (Hemiptera, Aphidiae)

机译：柠檬粉虱（Hemiptera，Aphidiae）在多种生物和非生物胁迫条件下定量PCR的参考基因验证

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The regulation of mRNA expression level is critical for gene expression studies. Currently, quantitative reverse transcription polymerase chain reaction (qRT-PCR) is commonly used to investigate mRNA expression level of genes under various experimental conditions. An important factor that determines the optimal quantification of qRT-PCR data is the choice of the reference gene for normalization. To advance gene expression studies in Toxoptera citricida (Kirkaldy), an important citrus pest and a main vector of the Citrus tristeza virus, we used five tools (GeNorm, NormFinder, BestKeeper, Delta Ct methods, and RefFinder) to evaluate seven candidate reference genes (elongation factor-1 alpha [EF1 alpha], beta tubulin [beta-TUB], 18S ribosomal RNA [18S], RNA polymerase II large subunit (RNAP II), beta actin (beta-ACT), alpha tubulin, and glyceraldhyde-3-phosphate dehydrogenase) under different biotic (developmental stages and wing dimorphism) and abiotic stress (thermal, starvation, and UV irradiation) conditions. The results showed that EF1 alpha and 18S were the most stable genes under various biotic states, beta-ACT and beta-TUB during thermal stress, EF1 alpha and RNAP II under starvation stress, and RNAP II, beta-ACT, and EF1 alpha under UV irradiation stress conditions. This study provides useful resources for the transcriptional profiling of genes in T. citricida and closely related aphid species.
机译：mRNA表达水平的调节对于基因表达研究至关重要。目前，定量逆转录聚合酶链反应（qRT-PCR）通常用于研究各种实验条件下基因的mRNA表达水平。确定qRT-PCR数据最佳定量的重要因素是参考基因的选择以进行标准化。为了在重要的柑橘害虫和柑橘柑橘的主要载体-柠檬粉虱（Toxoptera citricida，Kirkaldy）中进行基因表达研究，我们使用了五种工具（GeNorm，NormFinder，BestKeeper，Delta Ct方法和RefFinder）来评估七个候选参考基因（延伸因子1α[EF1α]，β微管蛋白[beta-TUB]，18S核糖体RNA [18S]，RNA聚合酶II大亚基（RNAP II），β肌动蛋白（beta-ACT），α微管蛋白和甘油醛- 3-磷酸​​脱氢酶）在不同的生物（发育阶段和机翼二态性）和非生物胁迫（热，饥饿和紫外线照射）下进行。结果表明，在各种生物状态下，EF1 alpha和18S是最稳定的基因，在热胁迫下是beta-ACT和beta-TUB，在饥饿胁迫下是EF1 alpha和RNAP II，在饥饿下是RNAP II，beta-ACT和EF1 alpha。紫外线照射应力条件。这项研究提供了有用的资源，用于在柠檬纤毛虫和紧密相关的蚜虫物种中进行基因转录谱分析。

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