title: Identification and Evaluation of Suitable Reference Genes for Normalization of MicroRNA Expression in Helicoverpa armigera (Lepidoptera: Noctuidae) Using Quantitative Real-Time PCR.
abstract: More and more studies have focused on microRNAs (miRNAs) expression in the pest Helicoverpa armigera (Lepidoptera: Noctuidae) recently. Quantitative real-time PCR (qRT-PCR) is being widely used in miRNA expression studies. Suitable reference genes are necessary for the correct analysis of results. In this study, 10 candidate genes of H. armigera were selected and analyzed for their expression stability under different biotic and abiotic conditions with 3 statistical methods, including geNorm, NormFinder, and Bestkeeper. Combination the best number of reference genes was calculated by geNorm. One target gene, let-7, was used to validate the selection of reference genes. The suitable candidate reference genes were shown as follows: miR-9 and U6 snRNA for developmental stages, miR-100 and U6 snRNA for larval tissues, miR-100 and miR-305 for adult tissues, miR-9 and miR-279 for parasitic treatment, miR-998 and U6 snRNA for nuclear polyhedrosis virus infection, miR-9 and U6 snRNA for insecticide treatment, miR-92a, miR-100, and miR-279 for temperature treatment, miR-92a, miR-305, and miR-998 for starvation treatment, miR-9 and miR-279 for light treatment, miR-305 and miR-998 for hormone treatment, and there was not one reference gene suitable for all samples. This study could promote future research on miRNAs expression in H. armigera with optimal reference genes under different experimental conditions.
authors.affiliation: Department of Entomology, China Agricultural University, Beijing 100094, China.
authors.forename: Yuhui
authors.initials: Y
authors.lastname: Yang
authors.affiliation: Department of Entomology, China Agricultural University, Beijing 100094, China.
authors.forename: Zhen
authors.initials: Z
authors.lastname: Li
authors.affiliation: Department of Entomology, China Agricultural University, Beijing 100094, China.|Department of Entomology, Henan Institute of Science and Technology, Xinxiang 453003, China.
authors.forename: Jinjun
authors.initials: J
authors.lastname: Cao
authors.affiliation: Department of Entomology, China Agricultural University, Beijing 100094, China.
authors.forename: Yanrong
authors.initials: Y
authors.lastname: Li
authors.affiliation: Department of Entomology, China Agricultural University, Beijing 100094, China.
authors.forename: Hui
authors.initials: H
authors.lastname: Li
authors.affiliation: Department of Entomology, China Agricultural University, Beijing 100094, China.
authors.forename: Qingpo
authors.initials: Q
authors.lastname: Yang
authors.affiliation: Department of Entomology, China Agricultural University, Beijing 100094, China.
authors.forename: Qingwen
authors.initials: Q
authors.lastname: Zhang
authors.affiliation: Department of Entomology, China Agricultural University, Beijing 100094, China.
authors.forename: Xiaoxia
authors.initials: X
authors.lastname: Liu
mesh_terms: D000818:Animals; D005260:Female; D020869:Gene Expression Profiling; D005786:Gene Expression Regulation; D007814:Larva / Q000235:genetics / Q000378:metabolism; D008297:Male; D035683:MicroRNAs / Q000378:metabolism*; D009036:Moths / Q000235:genetics* / Q000378:metabolism; D060888:Real-Time Polymerase Chain Reaction
chemical_list: D035683:MicroRNAs
keywords: cotton bollworm; microRNAs; qRT-PCR analysis; reference genes