RNA interference of a trehalose-6-phosphate synthase gene reveals its roles during larval-pupal metamorphosis in Bactrocera minax (Diptera: Tephritidae).
RNA interference of a trehalose-6-phosphate synthase gene reveals its roles during larval-pupal metamorphosis in Bactrocera minax (Diptera: Tephritidae).
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DOI：
10.1016/j.jinsphys.2016.07.003
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发表时间：
2016-08
影响因子：
2.2
通讯作者：
Ke-Cai Xiong;Jia Wang;Jia-hao Li;Yulu Deng;Po Pu;H. Fan;Ying-hong Liu
中科院分区：
文献类型：
--
作者：
Ke-Cai Xiong;Jia Wang;Jia-hao Li;Yulu Deng;Po Pu;H. Fan;Ying-hong Liu
海藻糖是昆虫体内的主要血糖，是昆虫体内的主要能量来源，也是几丁质生物合成的起始底物。在昆虫中，海藻糖是由一种重要的酶--海藻糖-6-磷酸合成酶（TPS）催化合成的。本研究克隆并鉴定了柑橘实蝇（Bactrocera minax）海藻糖-6-磷酸合酶基因（BmTPS），该基因的开放阅读框为2445个核苷酸，编码814个氨基酸，预测分子量为92.05 kDa，在柑橘实蝇各发育阶段均有表达，在末龄幼虫中表达量较高。组织特异性表达模式显示BmTPS主要在脂肪体中表达。20-羟基蜕皮激素（20 E）诱导BmTPS和几丁质合成途径中三个基因的表达。此外，将双链RNA注射到三龄幼虫中成功地沉默了BmTPSinB的转录。minax，从而降低TPS活性和海藻糖含量。此外，沉默BmTPsin抑制几丁质生物合成途径中的三个关键基因的表达，并表现出52%的死亡和异常表型。结果表明，BmTPS是幼虫-蛹变态所必需的。此外，还建立了B. minax为进一步研究该虫的分子生物学和生理学奠定了基础。
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 fromBactrocera minax(BmTPS) was cloned and characterized.BmTPScontained an open reading frame of 2445 nucleotides encoding a protein of 814 amino acids with a predicted molecular weight of 92.05 kDa.BmTPSwas detectable in all developmental stages ofBactrocera minaxand expressed higher in the final- (third-) instar larvae. Tissue-specific expression patterns ofBmTPSshowed that it was mainly expressed in the fat body. The 20-hydroxyecdysone (20E) induced the expression ofBmTPSand three genes in the chitin biosynthesis pathway. Moreover, injection of double-stranded RNA into third-instar larvae successfully silenced the transcription ofBmTPSinB. minax, and thereby decreased the activity of TPS and trehalose content. Additionally, silencing ofBmTPSinhibited the expression of three key genes in the chitin biosynthesis pathway and exhibited 52% death and abnormal phenotypes. The findings demonstrate thatBmTPSis indispensable for larval-pupal metamorphosis. Besides, the establishment of RNAi experimental system inB. minaxwould lay a solid foundation for further investigation of molecular biology and physiology of this pest.