黄龙病菌逃避柑橘木虱免疫细胞清除作用的分子机制研究

柑橘黄龙病是我国乃至全球柑橘类产业最重要的传染性病害，其病原是一种革兰氏阴性杆菌(Candidatus Liberibacter asiaticum，CLas)，由媒介昆虫柑橘木虱传播。本项目在探明柑橘木虱感染CLas后，CLas可进入柑橘木虱免疫细胞并在其内部长期存活的基础上。首次探究CLas如何调控柑橘木虱免疫细胞功能的分子机制，拟采用免疫荧光共定位和透射电镜技术，明确CLas在免疫细胞存活的具体细胞器位点；后利用细胞分离技术和超速离心法，分离纯化CLas存活的具体细胞器，并结合蛋白组和转录组学技术获取该细胞器组学信息；再通过数据分析筛选CLas调控柑橘木虱免疫细胞细胞器的差异表达蛋白或基因。揭示CLas成功存活于柑橘木虱免疫细胞内且无法被免疫细胞清除的分子机制，为后期建立CLas与柑橘木虱免疫细胞互作的研究体系以及未来通过调控柑橘木虱免疫系统来控制黄龙病的传播提供理论依据和技术保障。

Huanglongbing is an important infectious disease in the citrus industry of China and even the world. It is caused by a gram-negative of phloem-limited bacteria (Candidatus Liberibacter asiaticum, CLas), and is spread by its main vector insect, citrus psyllid. Preliminary experiments have found that CLas can survive in the immune cells of citrus psyllid for a long time after being infected. Therefore, this study took CLas and its vector insect citrus psyllid as the research object to explore how CLas regulates the function of cell immunity of citrus psyllids. This project intended to use co-immunofluorescence and transmission electron microscopy techniques to clarify the specific organelle sites where CLas survives in immune cells. Subsequently, the Percoll density gradient and sucrose gradient centrifugation were used to separate the organelle with CLas in immune cells. Then, through proteome and transcriptome analysis, the organelle information that were not infected and infected with CLas can be obtained. By comparing the two omics information, the interaction proteins and differentially expressed genes between CLas and the immune cells of citrus psyllid were screened. Finally, this study clarified the mechanism by which CLas survives in the immune cells of citrus psyllids. This research provides theoretical basis and technical support for the future elimination of CLas by regulating the citrus psyllid immune system, and control of the spread of huanglongbing, as well as the establish a research model for the interaction between CLas and immune cells of citrus psyllid.