Molecular characterization and functional analysis of USP - 1 by RNA interference in the Asian gypsy moth Lymantria disp
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ORIGINAL PAPER
Molecular characterization and functional analysis of USP-1 by RNA interference in the Asian gypsy moth Lymantria dispar Rongrong Wen1,2 • Qingquan Zhao1 • Buyong Wang2 • Yan Ma3 • Ling Ma1
Received: 11 April 2018 / Accepted: 20 September 2018 Northeast Forestry University 2019
Abstract Ecdysteroids play an important role in regulating diverse physiological processes in arthropods, such as molting, metamorphosis, reproduction and diapause. Ecdysteroids mediate the response by binding to a heterodimeric complex of two nuclear receptors: the ecdysone receptor and the ultraspiracle (USP). To investigate the role of USP in development of the Asian gypsy moth (Lymantria dispar), a USP cDNA was obtained from the transcriptome of L. dispar and verified by PCR. In-depth profiling of transcript levels of L. dispar USP-1 (LdUSP-1) at different developmental stages and over time in thirdinstar larvae and different tissues isolated during the thirdinstar stage of L. dispar was then carried out. Transcript levels of LdUSP-1 were relatively high before 72 h in the
Rongrong Wen, Qingquan Zhao, and Buyong Wang have contributed equally to this work. Project funding: This research was supported by Grants from the Natural Science Foundation of China (31770688), the Fundamental Research Funds for the Central Universities (2572017AA17), the Ph.D. Programs Foundation of HeZe University (XY18BS06), and the Ph.D. Programs Foundation of HeZe University (XY18BS05). The online version is available at http://www.springerlink.com Corresponding editor: Chai Ruihai. & Ling Ma [email protected] 1
School of Forestry, Northeast Forestry University, Harbin 150040, People’s Republic of China
2
College of Agricultural and Biological Engineering, Heze University, Heze 274015, People’s Republic of China
3
College of Management, Harbin University of Commerce, Harbin 150028, People’s Republic of China
third-instar larvae after ecdysis and in the adult male. The function of LdUSP-1 in molting was analyzed by knockdown of LdUSP-1 in third instar larvae using RNA interference. Silencing of LdUSP-1 significantly downregulated the transcript level of E75, an ecdysone-inducible gene, and of Sad, a Halloween gene. In addition, the duration of the third-instar stage was slightly shortened and larval mortality increased after the LdUSP-1 knockdown. Keywords Lymantria dispar Ultraspiracle gene Ecdysone RNA interference Development
Introduction In insects, ecdysteroids and juvenile hormones are important hormone families that regulate the complex processes of molting and metamorphosis. Ecdysteroids also coordinate many other aspects of insect life, such as reproduction, diapause and innate immunity (Raikhel et al. 2005; Verma and Tapadia 2015). Similar to other hormone systems, ecdysteroids autoregulate their own biosynthesis via direct and indirect feedback mechanisms (Parvy et al. 2014; Sakurai 2005). In insects, the ecdysteroid 20-hydroxyecdysone (20E) transduces its signal through a receptor heterodimer comprising two nuclear r
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