Function of the zasp52 gene detected by CRISPR/Cas9 in the global fruit borer Grapholita molesta

IF 3.7 2区 农林科学 Q2 BIOCHEMISTRY & MOLECULAR BIOLOGY
Sha Su , Zhimin Xu , Jinbo Suo , Yurong Zhou , Xiaohe Zhang , Amedius Dwigo Ignatus , Yayun Zuo , Xiong Peng , Fei Li , Maohua Chen
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Abstract

Zasp (Z band alternatively spliced PDZ-motif protein) is the core component of the Z-disc in muscle tissue and plays a vital role in the assembly and maintenance of myofibrils. The zasp has been studied in vertebrates, but it has only been reported in model organisms (e.g. Drosophila melanogaster) among insect species. Grapholita molesta is an important fruit pest with weak flight capacity. Flight capacity is important for the seasonal host switch of this pest. So far, the genes involved in the flight of G. molesta have not been analyzed. In this study, we identified and cloned the Gmzasp52 gene from G. molesta. This gene was expressed in different tissues and developmental stages of G. molesta. Using CRISPR/Cas9 gene editing technology, a homozygous Gmzasp52 gene knockout strain of G. molesta was successfully constructed. Knockout of the Gmzasp52 gene significantly prolonged the larval and preoviposition periods of G. molesta, and shortened the pupal period; the larval weight, pupal weight and fecundity decreased significantly, and the pupal mortality increased significantly; the Z-disc structure of the flight muscle was destroyed, and the myofibrils were damaged, resulting in a significant decrease in flight capacity. These results indicate that the Gmzasp52 plays an important role in the growth and development, reproduction, flight muscle structure and flight capacity of G. molesta. Our study reveals the role of zasp52 gene in a non-model insect species for the first time. The results not only analyzed the role of Gmzasp52 gene in G. molesta, but also further characterized the function of zasp52 gene in non-model organisms and provided possible target gene for the control of this pest.

Abstract Image

利用CRISPR/Cas9技术检测全球果螟zasp52基因的功能
Zasp (Z带选择性剪接pdz基序蛋白)是肌肉组织中Z盘的核心成分,在肌原纤维的组装和维持中起着至关重要的作用。zasp已经在脊椎动物中进行了研究,但在昆虫物种中仅在模式生物(如果蝇)中有报道。小蠹蛾是一种重要的水果害虫,其飞行能力较弱。飞行能力对该害虫的季节性寄主转换很重要。到目前为止,还没有分析过与毛蝇飞行有关的基因。在本研究中,我们鉴定并克隆了G. molesta Gmzasp52基因。该基因在不同组织和发育阶段均有表达。利用CRISPR/Cas9基因编辑技术,成功构建了G. molesta Gmzasp52基因纯合子敲除菌株。敲除Gmzasp52基因显著延长了小蠹蛾的幼虫期和产卵前期,缩短了蛹期;幼虫重、蛹重、繁殖力显著降低,蛹死亡率显著升高;飞行肌z盘结构被破坏,肌原纤维受损,飞行能力明显下降。上述结果表明,Gmzasp52基因在飞蛾的生长发育、繁殖、飞行肌肉结构和飞行能力等方面发挥着重要作用。我们的研究首次揭示了zasp52基因在非模式昆虫物种中的作用。研究结果不仅分析了Gmzasp52基因在鼠夜蛾中的作用,还进一步表征了zasp52基因在非模式生物中的功能,为鼠夜蛾的防治提供了可能的靶基因。
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来源期刊
CiteScore
7.40
自引率
5.30%
发文量
105
审稿时长
40 days
期刊介绍: This international journal publishes original contributions and mini-reviews in the fields of insect biochemistry and insect molecular biology. Main areas of interest are neurochemistry, hormone and pheromone biochemistry, enzymes and metabolism, hormone action and gene regulation, gene characterization and structure, pharmacology, immunology and cell and tissue culture. Papers on the biochemistry and molecular biology of other groups of arthropods are published if of general interest to the readership. Technique papers will be considered for publication if they significantly advance the field of insect biochemistry and molecular biology in the opinion of the Editors and Editorial Board.
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