Shen Xiu-Xian, Zhang Guo-Qiang, Zhao Yu-Xin, Zhu Xiao-Xiao, Yu Xiao-Fei, Yang Mao-Fa, Zhang Feng
Guizhou Provincial Key Laboratory for Agricultural Pest Management of the Mountainous Region, Institute of Entomology, College of Agriculture, Guizhou University, Guiyang, China.
Department of Entomology, College of Plant Protection, Nanjing Agricultural University, Nanjing, China.
Front Physiol. 2023 Oct 25;14:1277942. doi: 10.3389/fphys.2023.1277942. eCollection 2023.
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 (), , , , , , , and () 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 , , and were ranked as the best reference gene combinations for measuring gene expression levels among different developing stages and in various starvation treatments. and were recommended to normalize the gene expression levels among different adult tissues. , , and were recommended sugar-feeding conditions. To validate the utility of the selected reference pair, and , we estimated the tissue-biased expression level of a chemosensory protein gene (). As expected, 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 .
是一种捕食者，是用于控制农林蚜虫的重要生物因子。尽管许多研究已经调查了其生物学和生态学特征，但很少有分子研究报道。当前的研究旨在鉴定合适的内参基因，以通过定量逆转录PCR促进未来的基因表达和功能分析。选择了八个内参基因（）、、、、、、、和（）。使用qRT-PCR技术在四种不同的实验条件（发育阶段、成虫组织、糖处理和饥饿处理）下测定它们的表达水平。用五种方法（Ct值、geNorm、NormFinder、BestKeeper和RefFinder）评估稳定性。结果表明，、和被列为在不同发育阶段和各种饥饿处理中测量基因表达水平的最佳内参基因组合。和被推荐用于标准化不同成虫组织中的基因表达水平。、和被推荐用于糖喂养条件。为了验证所选内参基因对和的实用性，我们估计了一种化学感受蛋白基因（）的组织偏向性表达水平。正如预期的那样，在触角中高度表达，在腹部中低表达。这些发现将为未来对的分子生理学和生物化学研究奠定基础。
