小贯小绿叶蝉卵黄原蛋白基因（EonuVg）克隆和功能研究及其对吡虫啉的响应

摘要/Abstract

摘要： 为解析小贯小绿叶蝉（Empoasca onukii）卵黄原蛋白（Vitellogenin,Vg）的分子特征、表达模式及其对吡虫啉的响应机制,克隆了两条EonuVg全长cDNA序列,分别命名为EonuVg1（PP128347）和EonuVg2（PP128348）,其开放阅读框全长分别为6 066 bp和6 138 bp,分别编码2 021个和2 045个氨基酸。两者均含有信号肽及典型的Vg结构域,包括卵黄原蛋白N端（Vitellogenim-N superfamily,Vit-N）、卵黄原蛋白开放β-折叠（Vit_open_b-sht,Vit-b）和血管性血友病因子（Von Willebrand factor type D,VWD）结构域,并具有N端的多聚丝氨酸束、RNPR酶切位点、GLCG保守基序、C端的半胱氨酸残基等昆虫Vg中独特的氨基酸基序。RT-qPCR结果显示,EonuVg1和EonuVg2在小贯小绿叶蝉的卵和若虫发育前期表达量较低,表达峰值出现在初羽化雌虫阶段。组织表达分析显示,两个基因主要在脂肪体和卵巢中高表达。利用RNA干扰技术分别沉默两个基因后,尽管雌虫历期、产卵历期和产卵量均呈下降趋势,但与对照组相比差异不显著。吡虫啉胁迫试验结果表明,亚致死及以上浓度处理显著抑制了EonuVg1和EonuVg2的表达。该研究初步探讨了EonuVg在小贯小绿叶蝉发育及其对吡虫啉响应中的可能功能,为深入研究Vg的功能提供了理论基础和数据支撑。

关键词: 小贯小绿叶蝉, 卵黄原蛋白, 表达模式, 亚致死浓度

Abstract: This study systematically investigated the molecular characteristics, expression patterns of the vitellogenin (Vg) gene EonuVg, and its response to imidacloprid in Empoasca onukii. The full-length cDNA sequences of two EonuVg genes, EonuVg1 (PP128347) and EonuVg2 (PP128348) were cloned and designated. Their open reading frames (ORFs) are 6 066 bp and 6 138 bp, encoding 2 021 and 2 045 amino acids, respectively. Both proteins contain conserved structural domains of Vg, including a signal peptide, Vitellogenin-N superfamily (Vit-N), Vit_open_b-sht (Vit-b) and Von Willebrand factor type D (VWD) domains, along with insect-specific motifs such as N-terminal poly-serine tracts, an RNPR cleavage site, a GLCG conserved motif, and C-terminal cysteine residues. The RT-qPCR results show that EonuVg1 and EonuVg2 exhibited low expressions during the early developmental stages (eggs and nymphs), and their expression peaks appreared in the newly emerged female adults. Tissue-specific expression analysis shows that the two genes were highly expressed in the ovary and fat body. After silencing the two genes by RNAi technology, the female duration, oviposition duration and oviposition quantity show a downward trend, but there was no significant difference compared with the control group feeding with dsGFP. The results of imidacloprid stress experiment show that sublethal and higher concentrations of imidacloprid significantly downregulated the expressions of EonuVg1 and EonuVg2 in female adults. This study preliminarily discussed the possible function of EonuVgs in the development and their response to imidacloprid in E. onukii, which provided theoretical basis and data support for further study on the function of Vg genes.

Key words: Empoasca onukii, vitellogenin, expression pattern, sublethal concentration

中图分类号:

潘铖, 夏露霞, 魏可欣, 王梦馨, 钮羽群, 韩宝瑜. 小贯小绿叶蝉卵黄原蛋白基因（EonuVg）克隆和功能研究及其对吡虫啉的响应[J]. 茶叶科学, 2025, 45(3): 439-453.

PAN Cheng, XIA Luxia, WEI Kexin, WANG Mengxin, NIU Yuqun, HAN Baoyu. Cloning and Functional Analysis of Vitellogenin Genes (EonuVg) in Empoasca onukii and Their Response to Imidacloprid[J]. Journal of Tea Science, 2025, 45(3): 439-453.

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