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Validation and Evaluation of Reference Genes for Quantitative Real-Time PCR Analysis in Mythimna loreyi (Lepidoptera: Noctuidae)

文献类型： 外文期刊

作者： Wang, Liuyang 1 ; Yang, Chaoxia 2 ; Liu, Qingyu 2 ; Zhang, Xiaofang 1 ; Mei, Xiangdong 2 ; Zhang, Tao 1 ; Ning, Jun 2 ;

作者机构： 1.Minist Agr & Rural Affairs, HAAFS Key Lab IPM Crops Northern Reg North China, China IPM Innovat Ctr Hebei Prov, Plant Protect Inst,Int Sci & Technol Joint Res Ctr, Baoding 071000, Peoples R China

2.Chinese Acad Agr Sci, State Key Lab Biol Plant Dis & Insect Pests, Inst Plant Protect, Beijing 100193, Peoples R China

3.China Agr Univ, Coll Sci, Innovat Ctr Pesticide Res, Dept Appl Chem, Beijing 100193, Peoples R China

关键词： Mythimna loreyi; qRT-PCR; gene stability; reference genes; normalization

期刊名称：INSECTS （ 影响因子：3.0； 五年影响因子：3.1 ）

ISSN：

年卷期： 2024 年 15 卷 3 期

页码：

收录情况： SCI

摘要： Quantitative real-time PCR (qRT-PCR) is a widely applied technique for accurately assessing the expression of target genes. In practice, the evaluation of gene expression requires appropriate reference genes. To screen reliable reference genes for evaluating gene expression via qRT-PCR in Mythimna loreyi, a notorious migratory pest across Asia, Africa, Europe, and Australia, we assessed the expression stability of 13 candidate reference genes in M. loreyi using the Delta Ct method, BestKeeper, Normfinder, GeNorm, and the web-based comprehensive platform RefFinder. These reference genes include RPL10, RPL27, RPL32, RPS3, TATA-box, GAPDH, AK, Actin, EF, alpha-tubulin, SOD, 18S rRNA, and FTZ-F1, which is frequently employed in Lepidoptera insects. Our findings revealed that the performance of the candidate reference gene depended on experimental conditions. Specifically, RPL27 and RPL10 were the most suitable for evaluating expression changes across developmental stages, tissues, and adult ages. The optimal reference genes were recommended in specific experiment conditions, for instance, EF and RPS3 were recommended for mating status, AK and RPL10 were recommended for temperature treatments, RPL27 and FTZ-F1 were recommended for larva diet, and EF and RPL27 were recommended for adult diet treatments. Additionally, expression profiles of pheromone-binding protein 2 (MlorPBP2) and glutathione S-transferase (MlorGST1) were used to validate the reference genes. This study provides reference genes for the accurate normalization of qRT-PCR data, laying the groundwork for studying the expression of target genes in M. loreyi.

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