蚜虫是一种捕食者,是一种重要的生物制剂,用于控制农业和林业蚜虫。尽管许多研究已经调查了其生物学和生态学特征,很少有分子研究报道。进行当前研究以鉴定合适的参考基因,以通过定量逆转录PCR促进未来的基因表达和功能分析。8个参考基因甘油醛-3-磷酸脱氢酶(GAPDH),RPS13,RPL8,RPS3,α-浴缸,β-肌动蛋白,选择RPL32和延伸因子1α(EF1-α)。在四个不同的实验条件(发育阶段,成人组织,糖治疗,和饥饿处理)使用qRT-PCR技术。用五种方法评价稳定性(Ct值,geNorm,NormFinder,BestKeeper,和RefFinder)。结果表明,GAPDH,RPL32和EF1-α被列为测量不同发育阶段和各种饥饿处理中基因表达水平的最佳参考基因组合。建议使用RPL8和RPS3使不同成人组织中的基因表达水平正常化。RPL32,β-肌动蛋白,和EF1-α是推荐的糖饲喂条件。要验证所选引用对的实用程序,RPL8和RPS3,我们估计了化学感应蛋白基因(AaphCSP1)的组织偏向性表达水平。不出所料,AaphCSP1在触角中高表达,在腹部中低表达。这些发现将为今后开展蚜虫的分子生理和生物化学研究奠定基础。
Aphidoletes aphidimyza is a predator that is an important biological agent used to control agricultural and forestry aphids. Although many studies have investigated its biological and ecological characteristics, few molecular studies have been reported. The current study was performed to identify suitable reference genes to facilitate future gene expression and function analyses via quantitative reverse transcription PCR. Eight reference genes glyceraldehyde-3-phosphate dehydrogenase (GAPDH), RPS13, RPL8, RPS3, α-Tub, β-actin, RPL32, and elongation factor 1 alpha (EF1-α) were selected. Their expression levels were determined under four different experimental conditions (developmental stages, adult tissues, sugar treatment, and starvation treatment) using qRT-PCR technology. The stability was evaluated with five methods (Ct value, geNorm, NormFinder, BestKeeper, and RefFinder). The results showed that GAPDH, RPL32, and EF1-α were ranked as the best reference gene combinations for measuring gene expression levels among different developing stages and in various starvation treatments. RPL8 and RPS3 were recommended to normalize the gene expression levels among different adult tissues. RPL32, β-actin, and EF1-α were recommended sugar-feeding conditions. To validate the utility of the selected reference pair, RPL8, and RPS3, we estimated the tissue-biased expression level of a chemosensory protein gene (AaphCSP1). As expected, AaphCSP1 is highly expressed in the antennae and lowly expressed in the abdomen. These findings will lay the foundation for future research on the molecular physiology and biochemistry of A. aphidimyza.