Candidatus Liberibacter asiaticus exploits cytoskeletal system of psyllid vector for circulative propagative infection.

The citrus disease Huanglongbing (HLB) in Asia and the US is caused by Candidatus Liberibacter asiaticus (CLas), which is primarily transmitted by Diaphorina citri, also known as Asian citrus psyllid in a persistent and propagative manner. However, the exact mechanisms underlying CLas circulation within D. citri remain largely unclear. Here, immunofluorescence microscopy and electron microscopy were utilized to track the sequential infection of CLas in D. citri, from alimentary canal to salivary glands, and ultimately to the plant host. CLas was found to initially infect the epithelium of filter chamber, after which it rapidly spreads to visceral muscles for further infection throughout the alimentary canal. The rapid spread in D. citri adults causes the duration of CLas circulation to be as short as 9 days. The duration of latent period may be explained by the recruitment of cytoskeletal α-actinin by the outer membrane protein (OMP) of CLas. Inhibition of actin filament or knocking down the expression of α-actinin significantly suppresses CLas cytoskeleton-dependent infection in and spread among D. citri organs. Injection of prokaryotically expressed OMP into D. citri also recruits α-actinin, resembling the natural infection of CLas. Our studies showed that CLas exploits α-actinin and remolds actin machinery of D. citri for overcoming the midgut release barrier, facilitating its circulation in the vector. By shedding light on these mechanisms, this report reveals more detailed mechanisms in CLas infection in D. citri, and offers a plausible explanation for rapid dissemination of HLB in nature from the perspective of psyllid transmission.