昆虫学报 ›› 2025, Vol. 68 ›› Issue (8): 1050-1056.doi: 10.16380/j.kcxb.2025.08.003

• 研究论文 • 上一篇    下一篇

鹿眼蛱蝶浓核病毒结构蛋白基因vp1-4在草地贪夜蛾幼虫体内的表达动态分析

彭倩, 陈彦宁, 李如玉, 傅悦, 谌祖文*   

  1. (黄冈师范学院, 经济林木种质改良与资源综合利用湖北省重点实验室, 黄冈 438000)
  • 出版日期:2025-08-20 发布日期:2025-09-30

Dynamic analysis of expression of Junonia coenia densovirus structural protein genes vp1-4 in Spodoptera frugiperda (Lepidoptera: Noctuidae) larvae

PENG Qian, CHEN Yan-Ning, LI Ru-Yu, FU Yue, CHEN Zu-Wen*   

  1.  (Hubei Key Laboratory of Economic Forest Germplasm Improvement and Resources Comprehensive Utilization, Huanggang Normal University, Huanggang 438000, China)
  • Online:2025-08-20 Published:2025-09-30

摘要: 【目的】鹿眼蛱蝶浓核病毒(Junonia coenia densovirus, JcDV)通过漏扫描方式编码4个病毒结构蛋白(viral structural protein, VP) VP1, VP2, VP3和VP4。本研究旨在探究JcDV的4个结构蛋白基因(vp1-4)在草地贪夜蛾Spodoptera frugiperda幼虫体内的表达特征,为深入研究病毒结构蛋白VP1-4的功能及其组装机制提供依据。【方法】采用免疫荧光检测在转染pJcDV质粒的Hi5细胞内VP1-4的亚细胞定位;利用RT-PCR检测JcDV侵染草地贪夜蛾2龄幼虫后0, 3, 6, 12, 24, 48, 72, 96和120 h时vp1-4的表达量;采用Western blot检测JcDV侵染草地贪夜蛾2龄幼虫后0, 24, 48, 72, 96和120 h时VP1-4的表达量;用免疫组织化学技术检测JcDV侵染草地贪夜蛾2龄幼虫后0, 24和96 h时VP1-4的组织表达特性。【结果】VP1-4主要位于Hi5细胞质中,少量以多聚体形式分布于细胞核内; JcDV侵染草地贪夜蛾2龄幼虫后3 h时可检测到vp1-4的转录并至少持续至120 h; JcDV侵染后24-120 h时可检测到VP1-4的表达; JcDV侵染草地贪夜蛾2龄幼虫后96 h时在虫体的表皮和气管中可检测到VP1-4的表达,而肌肉和脂肪体中没有明显表达信号。【结论】JcDV在侵染草地贪夜蛾2龄幼虫后3和24 h时可分别检测到病毒结构蛋白基因vp1-4的转录和翻译,表明VP1-4可能在侵染早期已开始发挥重要功能; JcDV可侵染草地贪夜蛾幼虫的表皮和气管,可能是导致其死亡的原因之一。

关键词: 草地贪夜蛾, 鹿眼蛱蝶浓核病毒, 结构蛋白基因, 转录时相, 表达时相

Abstract: 【Aim】 Junonia coenia densovirus (JcDV) encodes four viral structural proteins, namely VP1, VP2, VP3 and VP4 via leaky scanning. This study aims to explore the expression characteristics of four viral structural protein genes (vp1-4) of JcDV in Spodoptera frugiperda larvae, so as to provide the basis for further studying on the function of the virus structural proteins VP1-4 and the mechanism of their assembly. 【Methods】 The subcellular localization of VP1-4 within Hi5 cells transfected with pJcDV plasmid was detected by immunofluorescence. The expression levels of vp1-4 in the 2nd instar larvae of S. frugiperda at 0, 3, 6, 12, 24, 48, 72, 96 and 120 h after infection by JcDV were analyzed through RT-PCR. The expression levels of VP1-4 in the 2nd instar larvae of S. frugiperda at 0, 24, 48, 72, 96 and 120 h after infection by JcDV were detected by Western blot. Immunohistochemistry was adopted to detect the tissue expression characteristics of VP1-4 in the 2nd instar larvae of S. frugiperda at 0, 24 and 96 h after infection by JcDV. 【Results】 VP1-4 were mainly located in the cytoplasm of Hi5 cells and only distributed in small amounts in the nucleus in the form of polymers. The transcription of vp1-4 could be detected in the 2nd instar larvae of S. frugiperda at 3 h after infection by JcDV, and stably extended to at least 120 h. VP1-4 were expressed at 24-120 h after infection by JcDV. The expression of VP1-4 was detected in the cuticle and trachea of the 2nd instar larvae of S. frugiperda at 96 h after infection by JcDV, however, there were no obvious expression signals in the muscle and fat body. 【Conclusion】 The transcription and translation of the viral structural protein genes vp1-4 could be detected in the 2nd instar larvae of S. frugiperda at 3 and 24 h, respectively, after infection by JcDV, indicating that VP1-4 might play an important role in the early stages of infection. JcDV could infect the cuticle and trachea of S. frugiperda larvae, which might be one of the causes accounting for their death.

Key words: Spodoptera frugiperdaJunonia coenia densovirus, structural protein gene, transcriptional phase, expression phase