Intracellular trafficking of begomoviruses in the midgut cells of their insect vector.

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Begomoviruses are exclusively transmitted by whiteflies in a persistent circulative manner and cause considerable economic losses to crop production worldwide. Previous studies have shown that begomoviruses accumulate in vesicle-like structures in whitefly midgut cells and that clathrin-mediated endocytosis is responsible for their internalization. However, the process by which begomoviruses are trafficked within whitefly midgut cells remains largely unknown. In this study, we investigated the roles of vesicle trafficking in the transport of Tomato yellow leaf curl virus (TYLCV), a begomovirus that has spread to over 50 countries and caused extensive damage to a range of important crops, within midgut cells of whitefly (Bemisia tabaci). By disrupting vesicle trafficking using RNA silencing and inhibitors, we demonstrated that the early steps of endosomal trafficking are important for the intracellular transport of TYLCV in the whitefly midgut. In addition, our data show that, unlike many animal viruses, TYCLV is trafficked within cells in a manner independent of recycling endosomes, late endosomes, lysosomes, the Golgi apparatus and the endoplasmic reticulum. Instead, our results suggest that TYLCV might be transported directly from early endosomes to the basal plasma membrane and released into the hemolymph. Silencing of the sorting nexin Snx12, which may be involved in membrane tubulation, resulted in fewer viral particles in hemolymph; this suggests that the tubular endosomal network may be involved in the transport of TYLCV. Our results also support a role for the endo-lysosomal system in viral degradation. We further showed that the functions of vector early endosomes and sorting nexin Snx12 are conserved in the transmission of several other begomoviruses. Overall, our data indicate the importance of early endosomes and the tubular endosomal network in begomovirus transmission.