Zhang Meng-Yi, Wei Dong, Li Ran, Jia Hong-Ting, Liu Yu-Wei, Taning Clauvis Nji Tizi, Wang Jin-Jun, Smagghe Guy
Key Laboratory of Entomology and Pest Control Engineering, College of Plant Protection, Southwest University, Chongqing, China.
Academy of Agricultural Sciences, Southwest University, Chongqing, China.
Arch Insect Biochem Physiol. 2018 Apr;97(4). doi: 10.1002/arch.21447. Epub 2018 Jan 23.
In insects, glutamine synthetase (GS), a key enzyme in the synthesis of glutamine, has been reported to be associated with embryonic development, heat shock response, and fecundity regulation. However, little is known about the influence of GS on postembryonic development. In this study, we demonstrate that blocking the activity of GS in the oriental fruit fly (Bactrocera dorsalis) with use of a GS-specific inhibitor (L-methionine S-sulfoximine), led to a significant delay in larval development, pupal weight loss, and inhibition of pupation. We further identify cloned and characterized two GS genes (BdGS-c and BdGS-m) from B. dorsalis. The two GS genes identified in B. dorsalis were predicted to be located in the cytosol (BdGS-c) and mitochondria (BdGS-m), and homology analysis indicated that both genes were similar to homologs from other Dipterans, such as Drosophila melanogaster and Aedes aegypti. BdGS-c was highly expressed in the larval stages, suggesting that cytosolic GS plays a predominant role in larval development. Furthermore, RNA interference experiments against BdGS-c, to specifically decrease the expression of cytosolic GS, resulted in delay in larval development as well as pupal weight loss. This study presents the prominent role played by BdGS-c in regulating larval development and suggests that the observed effect could have been modulated through ecdysteroid synthesis, agreeing with the reduced expression of the halloween gene spook. Also, the direct effects of BdGS-c silencing on B. dorsalis, such as larval lethality, delayed pupation, and late emergence, can be further exploited as novel insecticide target in the context of pest management.
在昆虫中,谷氨酰胺合成酶(GS)是谷氨酰胺合成中的关键酶,据报道其与胚胎发育、热休克反应和繁殖力调节有关。然而,关于GS对胚后发育的影响知之甚少。在本研究中,我们证明使用GS特异性抑制剂(L-蛋氨酸S-亚砜亚胺)阻断东方果实蝇(橘小实蝇)中GS的活性,会导致幼虫发育显著延迟、蛹体重减轻和化蛹受到抑制。我们进一步从橘小实蝇中克隆并鉴定了两个GS基因(BdGS-c和BdGS-m)。在橘小实蝇中鉴定出的这两个GS基因预计分别位于胞质溶胶(BdGS-c)和线粒体(BdGS-m)中,同源性分析表明这两个基因与其他双翅目昆虫(如黑腹果蝇和埃及伊蚊)的同源物相似。BdGS-c在幼虫阶段高度表达,表明胞质GS在幼虫发育中起主要作用。此外,针对BdGS-c的RNA干扰实验,以特异性降低胞质GS的表达,导致幼虫发育延迟以及蛹体重减轻。本研究揭示了BdGS-c在调节幼虫发育中所起的重要作用,并表明观察到的效应可能是通过蜕皮甾体合成来调节的,这与万圣节基因spook表达降低一致。此外,BdGS-c沉默对橘小实蝇的直接影响,如幼虫致死率、化蛹延迟和羽化延迟,在害虫管理背景下可进一步开发为新型杀虫剂靶标。