Yan Xin, Zhang Yibo, Xu Kangkang, Wang Yawei, Yang Wenjia
Guizhou Provincial Key Laboratory for Rare Animal and Economic Insect of the Mountainous Region, College of Biology and Environmental Engineering, Guiyang University, Guiyang 550005, China.
State Key Laboratory for Biology of Plant Diseases and Insect Pests, Institute of Plant Protection, Chinese Academy of Agricultural Sciences, Beijing 100193, China.
Insects. 2021 Jun 29;12(7):589. doi: 10.3390/insects12070589.
The tomato leaf miner, is a destructive pest of tomato. The leaf-mining activities of its larvae can cause significant yield losses. Real-time quantitative polymerase chain reaction (RT-qPCR) is commonly used to measure gene expression, and the selection of stable reference genes for calibration and standardization is critical for accurate use of RT-qPCR. We studied the stable expression of nine common housekeeping genes in . These were examined at different developmental stages, in larval tissues, as well as those induced by exposure to 20E and insecticides. Four dedicated algorithms (geNorm, BestKeeper, NormFinder, and ΔCt method) and online tool (RefFinder) were used to analyze and rank the tested reference genes. Based on the standardized gene expression data of target gene (), the applicability of specific reference genes was verified. The results clarify that the optimal internal reference genes vary greatly under different experimental conditions. and were the best reference genes for developmental stages; and for different tissues; and for 20E treatment; and for insecticide treatments. The most suitable reference genes in all experimental conditions are and .
番茄潜叶蛾是番茄的一种毁灭性害虫。其幼虫的潜叶活动会导致显著的产量损失。实时定量聚合酶链反应（RT-qPCR）常用于测量基因表达，选择稳定的内参基因进行校准和标准化对于准确使用RT-qPCR至关重要。我们研究了9种常见管家基因在番茄潜叶蛾中的稳定表达情况。这些基因在不同发育阶段、幼虫组织以及经20E和杀虫剂处理诱导的组织中进行了检测。使用四种专门算法（geNorm、BestKeeper、NormFinder和ΔCt法）以及在线工具（RefFinder）对测试的内参基因进行分析和排名。基于目标基因（）的标准化基因表达数据，验证了特定内参基因的适用性。结果表明，在不同实验条件下，最佳内参基因差异很大。和是发育阶段的最佳内参基因；和是不同组织的最佳内参基因；和是20E处理的最佳内参基因；和是杀虫剂处理的最佳内参基因。在所有实验条件下最合适的内参基因是和。
