Selection of reference genes for RT-qPCR analysis in Trichogramma chilonis (Hymenoptera: Trichogrammatidae)
文献类型： 外文期刊
作者： Xie, Lian-Cheng 1 ; Tian, Jun-Ce 2 ; Lu, Yan-Hui 2 ; Xu, Hong-Xing 2 ; Zang, Lian-Sheng 3 ; Lu, Zhong-Xian 2 ; Jin, Li 1 ;
作者机构： 1.Guizhou Univ, State Key Lab Breeding Base Green Pesticide & Agr, Key Lab Green Pesticide & Agr Bioengn, Minist Educ, Guiyang 550025, Peoples R China
2.Zhejiang Acad Agr Sci, Inst Plant Protect & Microbiol, State Key Lab Managing Biot & Chem Threats Qual &, 198 Shiqiao Rd, Hangzhou 310021, Peoples R China
3.Jilin Agr Univ, Engn Res Ctr Nat Enemy Insects, Inst Biol Control, Changchun 130118, Peoples R China
关键词： Trichogramma chilonis; Reference gene; RT-qPCR; Normalization
期刊名称：JOURNAL OF ASIA-PACIFIC ENTOMOLOGY （ 影响因子：1.303； 五年影响因子：1.427 ）
ISSN： 1226-8615
年卷期： 2021 年 24 卷 3 期
页码：
收录情况： SCI
摘要： Trichogramma chilonis is an important natural enemy for control of various Lepidoperan crop pests. The biology of T. chilonis is well-studied, but the molecular mechanisms of this biology require further study. Screening suitable reference genes is a vital step for use of RT-qPCR to understand underlying molecular physiology. In the present study, nine candidate reference genes including elongation factor 2 (EF2), ribosomal proteins (RPS23, RPL13, and RPL44), malate dehydrogenase (MDH), eukaryotic translation initiation factor 3 subunit F (EIF3F), zinc finger protein 268 (ZFP268), muscle specific protein 20 (MP20), and ATP synthase subunit alpha (ATP5F1A) were evaluated at different conditions including development stage, diet, temperature, and insecticide treatments. Four common algorithms (the Delta Ct method, geNorm, BestKeeper, and NormFinder) and RefFinder were used to analyze gene expression stability. Our results indicated that two reference genes used for normalization were sufficient, and the optimal combinations were: RPS23 and EF2 for developmental stages, ZFP268 and EF2 for feeding with different diets, ZFP268 and RPL13 for temperature treatments, and EF2 and RPL44 for insecticide treatments. The results provide preliminary determination of suitable reference gene for standard RT-qPCR analyses in T. chilonis, which might establish the foundation for further molecular biology research.
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