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RNA 干扰海藻糖-6-磷酸合酶基因揭示其在斜纹夜蛾(鳞翅目:夜蛾科)几丁质和脂类生物合成中的作用。

RNA interference of a trehalose-6-phosphate synthase gene reveals its roles in the biosynthesis of chitin and lipids in Heortia vitessoides (Lepidoptera: Crambidae).

机构信息

College of Forestry and Landscape Architecture, South China Agricultural University, Guangzhou, China.

出版信息

Insect Sci. 2020 Apr;27(2):212-223. doi: 10.1111/1744-7917.12650. Epub 2018 Dec 11.

DOI:10.1111/1744-7917.12650
PMID:30397994
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC7379938/
Abstract

Trehalose-6-phosphate synthase (TPS), an enzyme that hydrolyzes two glucose molecules to yield trehalose, plays a pivotal role in various physiological processes. In this study, we cloned the trehalose-6-phosphate synthase gene (HvTPS) and investigated its expression patterns in various tissues and developmental stages in Heortia vitessoides Moore (Lepidoptera: Crambidae). HvTPS was highly expressed in the fat body and after pupation or before molting. We knocked down TPS in H. vitessoides by RNA interference and found that 3.0 μg of dsHvTPS resulted in optimal interference at 24 h and 36 h post-injection and caused a sharp decline in the survival rate during the 5th instar larval-pupal stage and obviously abnormal or lethal phenotypes. Additionally, compared to the controls, TPS activity and trehalose contents were significantly lower and the glucose content was significantly higher 24 h or 36 h after injection with 3.0 μg of dsHvTPS. Furthermore, the silencing of HvTPS suppressed the expression of six key genes in the chitin biosynthesis pathway and one key gene related to lipid catabolism. The expression levels of two genes associated with lipid biosynthesis were upregulated. These results strongly suggest that HvTPS is essential for the normal growth and development of H. vitessoides and provide a reference for further studies of the utility of key genes involved in chitin and lipid biosynthesis for controlling insect development.

摘要

海藻糖-6-磷酸合酶(TPS)是一种能够水解两个葡萄糖分子生成海藻糖的酶,在各种生理过程中起着关键作用。本研究克隆了甜菜夜蛾(Heortia vitessoides Moore)海藻糖-6-磷酸合酶基因(HvTPS),并研究了其在不同组织和发育阶段的表达模式。HvTPS 在脂肪体中高度表达,在化蛹或蜕皮前表达。我们通过 RNA 干扰敲低了 H. vitessoides 中的 TPS,发现 3.0μg 的 dsHvTPS 在注射后 24 小时和 36 小时达到最佳干扰效果,并导致第 5 龄幼虫-蛹期的存活率急剧下降,表现出明显的异常或致死表型。此外,与对照组相比,注射 3.0μg 的 dsHvTPS 24 小时或 36 小时后,TPS 活性和海藻糖含量明显降低,葡萄糖含量明显升高。此外,HvTPS 的沉默抑制了甲壳质生物合成途径中 6 个关键基因和一个与脂类代谢相关的关键基因的表达。两个与脂质生物合成相关的基因的表达水平上调。这些结果强烈表明 HvTPS 对 H. vitessoides 的正常生长和发育至关重要,为进一步研究参与甲壳质和脂质生物合成的关键基因在昆虫发育控制中的应用提供了参考。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/3e6a/7379938/1a97b2424376/INS-27-212-g006.jpg
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https://cdn.ncbi.nlm.nih.gov/pmc/blobs/3e6a/7379938/95a614160fe0/INS-27-212-g001.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/3e6a/7379938/ee423cdd3617/INS-27-212-g002.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/3e6a/7379938/439c79b0f0f4/INS-27-212-g003.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/3e6a/7379938/11901e580f25/INS-27-212-g004.jpg
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