Du Hui, Wang Yu-Meng, Wang Xiao-Wei
Ministry of Agriculture Key Laboratory of Molecular Biology of Crop Pathogens and Insects, Zhejiang Key Laboratory of Biology and Ecological Regulation of Crop Pathogens and Insects, Institute of Insect Sciences, Zhejiang University, Hangzhou, China.
Methods Mol Biol. 2025;2912:35-47. doi: 10.1007/978-1-0716-4454-6_6.
Begomoviruses are exclusively transmitted by the whitefly Bemisia tabaci in a persistent circulative manner and pose serious threat to the production of important cash crops worldwide. Here, we describe the protocols to study how the whiteflies transmit begomoviruses, including maintenance of whitefly and species identification, preparation of tomato plants infected with virus, acquisition and transmission of virus by whiteflies, dissection of whitefly organs, analysis of virus quantity in whiteflies, immunofluorescence and fluorescence in situ hybridization (FISH) detection of virus in whitefly tissues, membrane feeding of double-stranded RNA (dsRNA) or antibody and microinjection of dsRNA in whiteflies.
双生病毒仅由烟粉虱以持久循环的方式传播，对全球重要经济作物的生产构成严重威胁。在此，我们描述了研究烟粉虱如何传播双生病毒的实验方案，包括烟粉虱的饲养与种类鉴定、感染病毒的番茄植株的准备、烟粉虱对病毒的获毒与传毒、烟粉虱器官解剖、烟粉虱体内病毒量分析、烟粉虱组织中病毒的免疫荧光和荧光原位杂交（FISH）检测、双链RNA（dsRNA）或抗体的膜饲以及dsRNA对烟粉虱的显微注射。
