Localization of Candidatus Liberibacter asiaticus, Associated with Citrus Huanglongbing Disease, in its Psyllid Vector using Fluorescence in situ Hybridization
Localization of Candidatus Liberibacter asiaticus, Associated with Citrus Huanglongbing Disease, in its Psyllid Vector using Fluorescence in situ Hybridization
复制标题
DOI：
10.1111/j.1439-0434.2011.01836.x
复制
发表时间：
2011-12-01
影响因子：
1.5
通讯作者：
Hall, David G.
中科院分区：
文献类型：
--
作者：
Ammar, El-Desouky;Shatters, Robert G., Jr.;Hall, David G.
黄龙病是世界上最具破坏性的柑橘病害之一，与黄龙病或柑橘绿化密切相关。Las由亚洲柑橘木虱（ACP）Diaphorina citri（半翅目：木虱科）以持续的方式传播，但其与木虱载体的相互作用，特别是在器官和细胞水平上，知之甚少。我们已经测试了几个荧光原位杂交（FISH）协议和三个核酸探针本地化的LAS在血淋巴涂片和解剖器官的ACP成人喂养的LAS感染的植物在现场或实验室和部分从LAS感染的柑橘叶。通过荧光原位杂交和激光共聚焦扫描显微镜在滤室、中肠、马氏管、血淋巴、唾液腺、卵巢和ACP的肌肉和脂肪组织中检测到Las，ACP从受感染的植物中获得Las，以及在受感染的柑橘叶的韧皮部中。Las在柑橘叶韧皮部和ACP血淋巴中表现为多形体或短细杆，其分散性和个体差异性更大，但在唾液腺和其他ACP器官和组织的细胞中更密集地聚集。我们的研究结果提供了第一次在原位演示拉斯维加斯感染木虱的各种器官和组织，并显示近系统感染的ACP拉斯维加斯。
The bacterium Candidatus Liberibacter asiaticus (Las) has been strongly associated with huanglongbing, or citrus greening, which is one of the most devastating citrus diseases worldwide. Las is transmitted by the Asian citrus psyllid (ACP) Diaphorina citri (Hemiptera: Psyllidae) in a persistent manner, but its interactions with the psyllid vector, particularly at the organ and cellular levels, are poorly understood. We have tested several fluorescence in situ hybridization (FISH) protocols and three nucleic acid probes for the localization of Las in haemolymph smears and dissected organs of ACP adults that fed on Las-infected plants in the field or laboratory and in sections from Las-infected citrus leaves. Las was detected by FISH and confocal laser scanning microscopy in the filter chamber, midgut, Malpighian tubules, haemolymph, salivary glands, ovaries and in muscle and fat tissues of ACP that acquired Las from infected plants, as well as in the phloem of infected citrus leaves. Las appeared as pleiomorphic bodies or short thin rods that were much more dispersed and individually distinct in citrus leaf phloem and in ACP haemolymph, but more densely aggregated in cells of the salivary glands and other ACP organs and tissues. Our results provide the first in situ demonstration of Las infection in various psyllid organs and tissues and show the near-systemic infection of ACP by Las.