亚洲柑橘木虱(ACP)DiaphorinacitriKuwayama是亚洲念珠菌(CLas)的主要载体,柑橘黄龙病(HLB)的致病原。ACP内部CLA的分布和动态对于理解传输如何,CLas的传播和感染在自然界中发生在其宿主载体内。在这项研究中,通过荧光原位杂交(FISH)和实时定量PCR(qPCR)技术检测了CLas在ACP5龄若虫和成虫各种组织中的分布和滴度变化。结果表明,以被感染的植物为食后,ACP5龄若虫和成虫100%被CLas感染,CLas在ACP的大多数组织中广泛分布。中肠内CLas的滴度,5龄若虫和成虫的唾液腺和血淋巴组织最高。与成年人相比,5龄若虫的这三个组织中的CLas滴度明显更高,在mycetome中,卵巢和睾丸明显低于成人。FISH可视化进一步证实了这些发现。对CLas的动态分析表明,它存在于ACP成年人的所有发育年龄中。在ACP成人的大多数组织中,随着年龄的增长,CLas的存在呈明显的上升趋势,包括中肠,血淋巴,唾液腺,脚,头部,角质层和肌肉。我们的发现对全面了解传播具有重要意义,CLas的传播和侵扰,这对于制定新的策略来阻止CLas的传播非常重要,因此有助于HLB的有效预防和控制。
The Asian citrus psyllid (ACP) Diaphorina citri Kuwayama is the leading vector of Candidatus Liberibacter asiaticus (CLas), the causative agent of citrus Huanglongbing (HLB) disease. The distribution and dynamics of CLas within ACP are critical to understanding how the transmission, spread and infection of CLas occurs within its host vector in nature. In this study, the distribution and titer changes of CLas in various tissues of ACP 5th instar nymphs and adults were examined by fluorescence in situ hybridization (FISH) and real-time quantitative PCR (qPCR) techniques. Results demonstrated that 100% of ACP 5th instar nymphs and adults were infected with CLas following feeding on infected plants, and that CLas had widespread distribution in most of the tissues of ACP. The titers of CLas within the midgut, salivary glands and hemolymph tissues were the highest in both 5th instar nymphs and adults. When compared with adults, the titers of CLas in these three tissues of 5th instar nymphs were significantly higher, while in the mycetome, ovary and testes they were significantly lower than those of adults. FISH visualization further confirmed these findings. Dynamic analysis of CLas demonstrated that it was present across all the developmental ages of ACP adults. There was a discernible upward trend in the presence of CLas with advancing age in most tissues of ACP adults, including the midgut, hemolymph, salivary glands, foot, head, cuticula and muscle. Our findings have significant implications for the comprehensive understanding of the transmission, dissemination and infestation of CLas, which is of much importance for developing novel strategies to halt the spread of CLas, and therefore contribute to the efficient prevention and control of HLB.