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利用 RNA 干扰技术对(鳞翅目:夜蛾科)基因的特征、敲除及亲本效应进行研究。

Characterization, Knockdown and Parental Effect of Gene of (Lepidoptera: Pyralidae) Revealed by RNA Interference.

机构信息

Provincial Key Laboratory for Agricultural Pest Management of Mountainous Regions, Institute of Entomology, Guizhou University, Guiyang, Guizhou 550025, China.

Department of Agronomy, Bahauddin Zakariya University, Multan 60800, Pakistan.

出版信息

Genes (Basel). 2020 Oct 26;11(11):1258. doi: 10.3390/genes11111258.

Abstract

Hexokinase (HK) is a key enzyme in chitin biosynthesis in insects and plays an important role in development and energy regulation. It also performs a crucial role in the synthesis of Glucose-6-phosphate and its putative functions are studied via injection of dsRNA corresponding to the gene from (). This study was designed to analyze the characteristics and expression patterns of HK-related genes in various tissues of at different developmental stages. The ORF is a 1359 bp in length, encoding a protein of 452 amino acids, with homology and cluster analysis showing that shares an 85.11% sequence similarity with hexokinase from was highly expressed in the ovary and in the fifth instar larvae. Injection of ds significantly suppressed mRNA expression (73.6%) 120 h post-dsRNA injection as compared to a control group. The results demonstrated an increased incidence of larval and pupal mortality of 80% and 78%, respectively, with significant variation in the sex ratio between males (68.33%) and females (35%), overt larval deformities, and a reduction in average weight gain observed 120 h post-dsRNA injection. In addition, ds-injected showed a significant reduction in ovulation per female and larval hatching rate, along with increased larval and pupal mortality and variation in male and female emergence over three generations (G1, G2, and G3). Taken together, the outcomes of the study provide a foundation to study gene function and a new dimension to control by transgenic RNAi technology.

摘要

己糖激酶(HK)是昆虫几丁质生物合成中的关键酶,在发育和能量调节中发挥重要作用。它在葡萄糖-6-磷酸的合成中也起着至关重要的作用,通过注射来自 ()的基因对应的 dsRNA 来研究其可能的功能。本研究旨在分析不同发育阶段的 不同组织中与 HK 相关的基因的特征和表达模式。该 ORF 长 1359bp,编码一个 452 个氨基酸的蛋白质,同源性和聚类分析表明,与来自 的己糖激酶具有 85.11%的序列相似性,在卵巢和第五龄幼虫中高度表达。与对照组相比,dsRNA 注射后 120 小时,mRNA 表达显著抑制(73.6%)。结果表明,幼虫和蛹的死亡率分别增加到 80%和 78%,雌雄比例有显著差异(雄性 68.33%,雌性 35%),明显的幼虫畸形,以及在 dsRNA 注射后 120 小时观察到的平均体重增加减少。此外,dsRNA 注射的 表现出每只雌虫产卵量和幼虫孵化率显著降低,同时幼虫和蛹死亡率增加,以及雄性和雌性出现的变化超过三代(G1、G2 和 G3)。综上所述,该研究的结果为研究基因功能提供了基础,并为通过转基因 RNAi 技术控制 提供了新的维度。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/d5dc/7693289/6b57ac407222/genes-11-01258-g001.jpg

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