草地贪夜蛾自噬相关基因的鉴定

马秋琴; 王禹生; 梁宽; 韩勇; 易辉玉; 邓小娟

doi:10.7671/j.issn.1001-411X.202004007

草地贪夜蛾自噬相关基因的鉴定

Identification of autophagy-related genes in fall armyworm, Spodoptera frugiperda

摘要

摘要:
目的利用草地贪夜蛾Spodoptera frugiperda基因组序列，鉴定草地贪夜蛾自噬相关基因(Autophagy-related gene, Atg)及其对病毒感染的自噬应答，为研究自噬的分子机制奠定基础。

方法对人类Homo spiens、果蝇Drosophila melanogaster等不同物种来源的自噬相关蛋白(ATG)序列进行相似性比对，搜索草地贪夜蛾自噬相关基因序列，并通过NCBI保守结构域搜索工具预测候选自噬相关蛋白中存在的功能结构域；并通过qRT-PCR检测重组病毒EGFP-AcMNPV感染后引起Sf9细胞中自噬相关基因的表达。

结果鉴定获得了草地贪夜蛾参与自噬体形成的核心Atg基因10多个，包括Atg3、Atg5、Atg6、Atg7、Atg10、Atg12、Atg13、Atg101的全序列以及Atg1、Atg2、Atg4、Atg9、Atg14、Atg16、Atg17、Atg18的部分序列，Atg2、Atg4、Atg5、Atg6、Atg7、Atg8、Atg12等自噬相关基因在重组病毒EGFP-AcMNPV感染Sf9细胞6~12 h后表达量上调。

结论获得草地贪夜蛾基因组中真核生物保守的自噬相关基因，暗示草地贪夜蛾的自噬途径在进化中是保守的。AcMNPV病毒感染Sf9细胞可能引发宿主细胞的自噬响应。

Abstract:
Objective To identify autophagy-related genes (Atg) in fall armyworm (Spodoptera frugiperda) genome and their response to viral infection in S. frugiperda, and provide a basis for further research on the molecular mechanism of autophagy.

Method Autophagy-related protein (ATG) sequences from various species, such as human (Homo sapiens) and fruit fly (Drosophila melanogaster), were aligned and analyzed for sequence similarity. ATG sequences in S. frugiperda genome were searched. Conservative domain (CD) of the candidate ATG were analyzed by NCBI CD search. Furthermore, qRT-PCR was used to detect the expression changes of Atgs in Sf9 cells after infection by recombinant EGFP-AcMNPV.

Result We identified more than ten core Atgs which were involved in autophagosome formation in S. frugiperda, including the complete sequences of Atg3, Atg5, Atg6, Atg7, Atg10, Atg12, Atg13 and Atg101, and the partial sequences of Atg1, Atg2, Atg4, Atg9, Atg14, Atg16, Atg17 and Atg18. The expression levels of S. frugiperda Atg2, Atg4, Atg5, Atg6, Atg7, Atg8 and Atg12 were up-regulated after 6 to 12 hours infection by EGFP-AcMNPV.

Conclusion Eukaryotic conserved Atgs exist in the genome of S. frugiperda, indicating the autophagy pathway in S. frugiperda is evolutionarily conserved. AcMNPV infection might trigger the autophagic response in Sf9 cells.

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