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RNA interference-mediated knockdown of the Halloween gene Spookiest (CYP307B1) impedes adult eclosion in the western tarnished plant bug, Lygus hesperus.
Insect Mol Biol. 2016 10; 25(5):550-65.IM

Abstract

Ecdysteroids play a critical role in coordinating insect growth, development and reproduction. A suite of cytochrome P450 monooxygenases coded by what are collectively termed Halloween genes mediate ecdysteroid biosynthesis. In this study, we describe cloning and RNA interference (RNAi)-mediated knockdown of the CYP307B1 Halloween gene (Spookiest) in the western tarnished plant bug, Lygus hesperus. Transcripts for Ly. hesperus Spookiest (LhSpot) were amplified from all life stages and correlated well with timing of the pre-moult ecdysteroid pulse. In adults, LhSpot was amplified from heads of both genders as well as female reproductive tissues. Heterologous expression of a LhSpot fluorescent chimera in cultured insect cells co-localized with a fluorescent marker of the endoplasmic reticulum/secretory pathway. RNAi-mediated knockdown of LhSpot in fifth instars reduced expression of ecdysone-responsive genes E74 and E75, and prevented adult development. This developmental defect was rescued following application of exogenous 20-hydroxyecdysone but not exogenous 7-dehydrocholesterol. The unequivocal RNAi effects on Ly. hesperus development and the phenotypic rescue by 20-hydroxyecdysone are causal proof of the involvement of LhSpot in ecdysteroid biosynthesis and related developmental processes, and may provide an avenue for development of new control measures against Ly. hesperus.

Authors+Show Affiliations

USDA-ARS Arid Land Agricultural Research Center, Maricopa, AZ, USA.Zhejiang University, Hangzhou, China.Zhejiang University, Hangzhou, China.USDA-ARS Arid Land Agricultural Research Center, Maricopa, AZ, USA.USDA-ARS Arid Land Agricultural Research Center, Maricopa, AZ, USA.

Pub Type(s)

Journal Article
Research Support, Non-U.S. Gov't

Language

eng

PubMed ID

27189651

Citation

Van Ekert, E, et al. "RNA Interference-mediated Knockdown of the Halloween Gene Spookiest (CYP307B1) Impedes Adult Eclosion in the Western Tarnished Plant Bug, Lygus Hesperus." Insect Molecular Biology, vol. 25, no. 5, 2016, pp. 550-65.
Van Ekert E, Wang M, Miao YG, et al. RNA interference-mediated knockdown of the Halloween gene Spookiest (CYP307B1) impedes adult eclosion in the western tarnished plant bug, Lygus hesperus. Insect Mol Biol. 2016;25(5):550-65.
Van Ekert, E., Wang, M., Miao, Y. G., Brent, C. S., & Hull, J. J. (2016). RNA interference-mediated knockdown of the Halloween gene Spookiest (CYP307B1) impedes adult eclosion in the western tarnished plant bug, Lygus hesperus. Insect Molecular Biology, 25(5), 550-65. https://doi.org/10.1111/imb.12242
Van Ekert E, et al. RNA Interference-mediated Knockdown of the Halloween Gene Spookiest (CYP307B1) Impedes Adult Eclosion in the Western Tarnished Plant Bug, Lygus Hesperus. Insect Mol Biol. 2016;25(5):550-65. PubMed PMID: 27189651.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - RNA interference-mediated knockdown of the Halloween gene Spookiest (CYP307B1) impedes adult eclosion in the western tarnished plant bug, Lygus hesperus. AU - Van Ekert,E, AU - Wang,M, AU - Miao,Y-G, AU - Brent,C S, AU - Hull,J J, Y1 - 2016/05/18/ PY - 2016/5/19/entrez PY - 2016/5/18/pubmed PY - 2017/9/8/medline KW - CYP307B1 KW - Halloween gene KW - Lygus hesperus KW - RNA interference KW - Spookiest KW - western tarnished plant bug SP - 550 EP - 65 JF - Insect molecular biology JO - Insect Mol Biol VL - 25 IS - 5 N2 - Ecdysteroids play a critical role in coordinating insect growth, development and reproduction. A suite of cytochrome P450 monooxygenases coded by what are collectively termed Halloween genes mediate ecdysteroid biosynthesis. In this study, we describe cloning and RNA interference (RNAi)-mediated knockdown of the CYP307B1 Halloween gene (Spookiest) in the western tarnished plant bug, Lygus hesperus. Transcripts for Ly. hesperus Spookiest (LhSpot) were amplified from all life stages and correlated well with timing of the pre-moult ecdysteroid pulse. In adults, LhSpot was amplified from heads of both genders as well as female reproductive tissues. Heterologous expression of a LhSpot fluorescent chimera in cultured insect cells co-localized with a fluorescent marker of the endoplasmic reticulum/secretory pathway. RNAi-mediated knockdown of LhSpot in fifth instars reduced expression of ecdysone-responsive genes E74 and E75, and prevented adult development. This developmental defect was rescued following application of exogenous 20-hydroxyecdysone but not exogenous 7-dehydrocholesterol. The unequivocal RNAi effects on Ly. hesperus development and the phenotypic rescue by 20-hydroxyecdysone are causal proof of the involvement of LhSpot in ecdysteroid biosynthesis and related developmental processes, and may provide an avenue for development of new control measures against Ly. hesperus. SN - 1365-2583 UR - https://www.unboundmedicine.com/medline/citation/27189651/RNA_interference_mediated_knockdown_of_the_Halloween_gene_Spookiest__CYP307B1__impedes_adult_eclosion_in_the_western_tarnished_plant_bug_Lygus_hesperus_ L2 - https://doi.org/10.1111/imb.12242 DB - PRIME DP - Unbound Medicine ER -