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Li Feifei,Li Yan,Wang Tao,Li Tianfeng,Zhu Shuying,Pei Pei,Tang Guanghui 한국곤충학회 2020 Entomological Research Vol.50 No.6
Atrijuglans hetaohei Yang (Lepidoptera: Gelechioidea), is one of the major pests that seriously damage the walnut fruits. Although the morphology and physiology of A. hetaohei have been widely studied, suitable reference genes for normalizing target gene expression have not been identified. In this study, the expressions of eight candidate reference genes including GAPDH, β-Tubulin, TATA-binding protein (TBP), Histone, Ubiquitin-conjugating enzyme 9 (UBC9), arginine kinase (AK), 28S and elongation factor 1 alpha (EF1α) in different developmental stages and various larval tissues of A. hetaohei were determined using reverse transcription-quantitative polymerase chain reaction. Moreover, the expression stability of the selected reference genes was evaluated by ΔCt method, BestKeeper, NormFinder, geNorm and ReFinder, a comprehensive software platform. These results demonstrated that the best reference genes were GAPDH and 28S at different developmental stages of A. hetaohei; and the optimum references in various larval tissues of A. hetaohei were 28S, Histone and TBP. Our study should be useful to analyze the expression profiles of target genes and form a solid foundation for future research on the understanding of the genes’ biological functions in A. hetaohei.
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Zhengqing Zhang,Tianfeng Li,Guanghui Tang 한국응용곤충학회 2018 Journal of Asia-Pacific Entomology Vol.21 No.1
Atrijuglans hetaohei Yang (Lepidoptera: Gelechioidea) is a kind of walnut pest, which mainly damage Juglans regia(Juglandales: Juglandaceae) and is widely distributed in China. The information of microRNAs, e.g., the conservedand novel miRNAs of this species is still unknown. In the present study, the miRNAs in larval and adultstages of A. hetaohei were identified by high throughput sequencing with reference to the genome database of theBombyx mori. A total of 132 conserved miRNAs belonging to 64 families and seven novel miRNAs were obtained,among which 118 miRNAs were identified in both libraries, 4 and 17 miRNAs were only detected to adultspecificand larval-specific libraries, respectively. The most abundant miRNAs in A. hetaohei were miR-1a-3p, let-7, miR-2766 and miR-184. Comparative analysis revealed that 4 novel miRNAs and 47 conserved miRNAs weredifferentially expressed (P < 0.05) between two libraries. Abundance changes of miRNAs were observed fromlarva to adult: 19 miRNAs were up-regulated and 21 miRNAs were down-regulated. The metamorphosis andapoptosis associated miRNAs including miR-1a-3p, miR-133, let-7, miR-2766, miR-184 and miR-34 were discussedfor their predicted functions in insect development. The different expressions of these miRNAs wererelevant to dorso-ventral axis formation, phototransduction, insect hormone biosynthesis, circadian rhythm andmitogen-activated protein kinase (MAPK) signaling pathway. qRT-PCR analysis of ten miRNAs showed similarexpression patterns as high throughput analysis, most miRNAs were highly expressed in adult stage, while miR-8-3p, miR-305-3p and miR-2766 were highly expressed in larvae. The results preliminarily revealed the characterizationof conserved and novel miRNAs in different development stages of A. hetaohei. The different expressionsand associated functions of these miRNAs will provide a basis for understanding post-transcriptionallevel during A. hetaohei development and growth.
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