Wang Zhengbing, Zhang Hongfei, Zhang Ziyi, Zhao Jiayu, Ma Feilong, Zheng Manman, Yang Mingsheng, Sang Xiaoyue, Ma Keshi, Li Lili
College of Life Science and Agronomy, Zhoukou Normal University, Zhoukou, China.
Key Laboratory of Plant Genetics and Molecular Breeding, Zhoukou Normal University, Zhoukou, China.
J Econ Entomol. 2022 Dec 14;115(6):2083-2091. doi: 10.1093/jee/toac162.
The soybean aphid Aphis glycines Matsumura is a predominant insect pest in Asia and North America and causes great losses to soybean. The release of genome data for A. glycines will facilitate gene function research in the future. However, suitable reference genes for A. glycines under various experimental conditions are scarce. To search for appropriate reference genes for A. glycines, nine candidate reference genes, including Act, α-Tub, β-Tub, RPS12, RPS18, RPL5, RPL27, EF1α, and Fer, were tested under six experimental conditions to evaluate their suitability for use in the normalization of qRT‒PCR data. Results showed that EF1α and RPS12 were optimal for the developmental stages of A. glycines, RPS18 and RPS12 were appropriate for wing dimorphism, β-Tub and RPS18 were suitable for different tissues and RPL5, and α-Tub could be used for normalization at different temperatures. β-Tub and EF1α could be proposed as reference genes for insecticide treatment, and RPL5 and RPS12 were found to be the most stable reference genes in different photoperiods. The results provide appropriate reference genes for analyzing gene expression in A. glycines and contribute to future research on the molecular physiology and biochemistry of A. glycines.
大豆蚜Aphis glycines Matsumura是亚洲和北美的主要害虫，给大豆造成了巨大损失。大豆蚜基因组数据的发布将有助于未来的基因功能研究。然而，在各种实验条件下适合大豆蚜的内参基因却很匮乏。为了寻找适合大豆蚜的内参基因，对包括Act、α-Tub、β-Tub、RPS12、RPS18、RPL5、RPL27、EF1α和Fer在内的9个候选内参基因在6种实验条件下进行了测试，以评估它们在定量逆转录聚合酶链反应（qRT-PCR）数据标准化中的适用性。结果表明，EF1α和RPS12在大豆蚜发育阶段表现最佳，RPS18和RPS12适用于翅型二态性，β-Tub和RPS18适用于不同组织以及RPL5，α-Tub可用于不同温度下的标准化。β-Tub和EF1α可被提议作为杀虫剂处理的内参基因，RPL5和RPS12被发现是不同光周期下最稳定的内参基因。这些结果为分析大豆蚜基因表达提供了合适的内参基因，并有助于未来对大豆蚜分子生理学和生物化学的研究。
