title: RNA interference of a trehalose-6-phosphate synthase gene reveals its roles during larval-pupal metamorphosis in Bactrocera minax (Diptera: Tephritidae).
abstract: Trehalose is the major blood sugar in insects, which plays a crucial role as an instant source of energy and the starting substrate for chitin biosynthesis. In insects, trehalose is synthesized by catalysis of an important enzyme, trehalose-6-phosphate synthase (TPS). In the present study, a trehalose-6-phosphate synthase gene from Bactrocera minax (BmTPS) was cloned and characterized. BmTPS contained an open reading frame of 2445 nucleotides encoding a protein of 814 amino acids with a predicted molecular weight of 92.05kDa. BmTPS was detectable in all developmental stages of Bactrocera minax and expressed higher in the final- (third-) instar larvae. Tissue-specific expression patterns of BmTPS showed that it was mainly expressed in the fat body. The 20-hydroxyecdysone (20E) induced the expression of BmTPS and three genes in the chitin biosynthesis pathway. Moreover, injection of double-stranded RNA into third-instar larvae successfully silenced the transcription of BmTPS in B. minax, and thereby decreased the activity of TPS and trehalose content. Additionally, silencing of BmTPS inhibited the expression of three key genes in the chitin biosynthesis pathway and exhibited 52% death and abnormal phenotypes. The findings demonstrate that BmTPS is indispensable for larval-pupal metamorphosis. Besides, the establishment of RNAi experimental system in B. minax would lay a solid foundation for further investigation of molecular biology and physiology of this pest.
authors.affiliation: Institute of Entomology, College of Plant Protection, Southwest University, Chongqing 400716, China.
authors.forename: Ke-Cai
authors.initials: KC
authors.lastname: Xiong
authors.affiliation: Institute of Entomology, College of Plant Protection, Southwest University, Chongqing 400716, China.
authors.forename: Jia
authors.initials: J
authors.lastname: Wang
authors.affiliation: Institute of Entomology, College of Plant Protection, Southwest University, Chongqing 400716, China.
authors.forename: Jia-Hao
authors.initials: JH
authors.lastname: Li
authors.affiliation: National Citrus Virus Exclusion Center, Citrus Research Institute, Southwest University, Chongqing 400712, China.
authors.forename: Yu-Qing
authors.initials: YQ
authors.lastname: Deng
authors.affiliation: Institute of Entomology, College of Plant Protection, Southwest University, Chongqing 400716, China.
authors.forename: Po
authors.initials: P
authors.lastname: Pu
authors.affiliation: Institute of Entomology, College of Plant Protection, Southwest University, Chongqing 400716, China.
authors.forename: Huan
authors.initials: H
authors.lastname: Fan
authors.affiliation: Institute of Entomology, College of Plant Protection, Southwest University, Chongqing 400716, China. Electronic address: yhliu@swu.edu.cn.
authors.forename: Ying-Hong
authors.initials: YH
authors.lastname: Liu
mesh_terms: D000818:Animals; D004441:Ecdysterone / Q000378:metabolism; D015870:Gene Expression; D005964:Glucosyltransferases / Q000235:genetics* / Q000378:metabolism; D019476:Insect Proteins / Q000235:genetics* / Q000378:metabolism; D007814:Larva / Q000235:genetics / Q000254:growth & development / Q000378:metabolism; D008675:Metamorphosis, Biological; D011679:Pupa / Q000235:genetics / Q000254:growth & development / Q000378:metabolism; D034622:RNA Interference; D033621:Tephritidae / Q000235:genetics / Q000254:growth & development* / Q000378:metabolism
chemical_list: D019476:Insect Proteins; D004441:Ecdysterone; D005964:Glucosyltransferases; C023180:trehalose-6-phosphate synthase
keywords: 20-Hydroxyecdysone; Bactrocera minax; Chitin biosynthesis; Insect metamorphosis; RNA interference; Trehalose-6-phosphate synthase