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钟嘉怡, 李莉, 王锡锋, 刘文文*.灰飞虱冠蛋白的基因克隆与表达分析[J].植物保护,2024,50(2):178-185.
灰飞虱冠蛋白的基因克隆与表达分析
Gene cloning and expression analysis of coronins in Laodelphax striatellus
投稿时间:2023-02-22  修订日期:2023-03-06
DOI:10.16688/j.zwbh.2023077
中文关键词:  灰飞虱  水稻条纹病毒  冠蛋白  转录水平
英文关键词:Laodelphax striatellus  rice stripe tenuivirus  coronin  transcription level
基金项目:国家自然科学基金(32270173); 国家重点研发计划政府间国际科技创新合作重点专项(2019YFE0108500)
作者单位E-mail
钟嘉怡, 李莉, 王锡锋, 刘文文* 中国农业科学院植物保护研究所, 植物病虫害综合治理全国重点实验室, 北京100193 liuwenwen01@caas.cn 
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中文摘要:
      灰飞虱Laodelphax striatellus冠蛋白属于WD40重复超家族成员,主要通过结合肌动蛋白调控细胞骨架的动态聚合与解聚。本研究通过RT-PCR克隆并获得了灰飞虱3个冠蛋白coronin 1A、coronin 2B和coronin 7的基因CDS区。系统进化树分析显示,这3个冠蛋白分别属于3个分支,且与褐飞虱Nilaparvata lugens的3个冠蛋白同源性最高,氨基酸序列相似性分别为99.2%、88.2%和82.5%。荧光定量PCR检测结果显示,coronin 1A在成虫中的转录水平显著高于2龄、4龄若虫,coronin 2B在雄虫中的转录水平显著高于2龄、4龄若虫和雌虫,coronin 7在雌虫中的转录水平显著高于2龄、4龄若虫和雄虫。而3个冠蛋白基因在唾液腺中的转录水平都显著高于肠道。灰飞虱携带水稻条纹病毒(rice stripe tenuivirus, RSV)后体内3个冠蛋白基因的转录水平显著高于对照,说明冠蛋白可能在灰飞虱传播RSV过程中发挥一定作用,研究结果为深入探究冠蛋白的功能奠定了基础。
英文摘要:
      The coronins of Laodelphax striatellus are the members of WD40-repeat protein superfamily, which can regulate the dynamic polymerization and depolymerization of actin cytoskeleton. In this study, we cloned the coding sequences of three coronins: coronin 1A, coronin 2B, and coronin 7 of L. striatellus by RT-PCR. Phylogenetic analysis showed that these three coronins belonged to separate branches, with the highest homology observed with Nilaparvata lugens, showing amino acid sequence similarities of 99.2%, 88.2% and 82.5%, respectively. Analysis of transcription levels of coronin genes showed that transcription level of coronin 1A in adults was significantly higher than that in the 2nd- and 4th-instar nymphs, transcription level of coronin 2B in male adults was significantly higher than in the 2nd- and 4th-instar nymphs and female adults, and transcription level of coronin 7 in female adults was significantly higher than that in the 2nd- and 4th-instar nymphs and male adults. Moreover, the transcription level of the three coronin genes in the salivary glands were significantly higher than those in the gut. In addition, the transcription levels of the three coronin genes in L. striatellus carrying rice stripe tenuivirus (RSV) were significantly higher than in non-viruliferous insects, indicating that the three coronins may play a role in transmitting RSV. This study have laid a foundation for further studies on the function of coronins in virus-transmission.
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