Galetto Luciana, Vallino Marta, Rashidi Mahnaz, Marzachì Cristina
Istituto per la Protezione Sostenibile delle Piante, CNR, Torino, Italy.
Citrus Experimental Station, University of Florida, Lake Alfred, FL, USA.
Methods Mol Biol. 2019;1875:307-317. doi: 10.1007/978-1-4939-8837-2_23.
To visualize phytoplasmas at early stages of vector infection, an immunofluorescence assay was developed. The chapter provides experimental details on dissection of salivary glands, incubation of the dissected organs with phytoplasma suspension, fixation, embedding, sectioning, labeling, and final visualization with confocal microscopy. All the procedure will be described for the leafhopper Euscelidius variegatus, natural vector of "Candidatus phytoplasma asteris" and laboratory vector of Flavescence dorée phytoplasma.
为了在媒介感染的早期阶段观察植原体，开发了一种免疫荧光测定法。本章提供了有关唾液腺解剖、将解剖后的器官与植原体悬浮液孵育、固定、包埋、切片、标记以及最后用共聚焦显微镜观察的实验细节。所有步骤将针对叶蝉Euscelidius variegatus进行描述，它是“Ca. phytoplasma asteris”的自然媒介和葡萄黄化植原体的实验室媒介。
