A vector whitefly endocytic receptor facilitates the entry of begomoviruses into its midgut cells via binding to virion capsid proteins.

Many circulative plant viruses transmitted by insect vectors are devastating to agriculture worldwide. The midgut wall of vector insects represents a major barrier and at the same time the key gate a circulative plant virus must cross for productive transmission. However, how these viruses enter insect midgut cells remains poorly understood. Here, we identified an endocytic receptor complex for begomoviruses in the midgut cells of their whitefly vector. Our results show that two whitefly proteins, BtCUBN and BtAMN, compose a receptor complex BtCubam, for which BtCUBN contributes a viral-binding region and BtAMN contributes to membrane anchorage. Begomoviruses appear to be internalized together with BtCubam via its interaction with the 12-19 CUB domains of BtCUBN via clathrin-dependent endocytosis. Functional analysis indicates that interruption of BtCUBN and BtAMN lead to reduction of virus acquisition and transmission by whitefly. In contrast, CUBN-begomovirus interaction was not observed in two non-competent whitefly-begomovirus combinations. These observations suggest a major role of the specific endocytic receptor in facilitating viral entry into vector midgut cells.

Implication of whitefly vesicle associated membrane protein-associated protein B in the transmission of Tomato yellow leaf curl virus.

Tomato yellow leaf curl virus (TYLCV) poses serious threat to tomato production worldwide, and the vector, Bemisia tabaci, plays a key role in the transmission of this virus. However, the molecular mechanisms underlying the transmission remain poorly understood. In this study, firstly, we identified the whitefly proteins that presumably interact with TYLCV coat protein (CP) using split-ubiquitin yeast two-hybrid system. Next, we conducted GST pull-down and immunofluorescence to examine the potential interaction between TYLCV CP and one of the proteins identified, namely vesicle associated membrane protein-associated protein B (VAPB), an protein abundantly expressed in whitefly midgut. Further experiments demonstrated that VAPB was significantly up-regulated upon virus acquisition, and silencing VAPB led to a significant increase of relative virus quantity in whitefly haemolymph and salivary glands, as well as an increase of TYLCV transmission efficiency. These findings indicate an important role of VAPB in the transmission of TYLCV by whiteflies.

Comparative proteomic analysis provides new insight into differential transmission of two begomoviruses by a whitefly.

BACKGROUND: Viruses in the genus Begomovirus (Family Geminiviridae) include many important economic plant viruses transmitted by whiteflies of the Bemisia tabaci species complex. In general, different begomoviruses may be acquired and transmitted by the same whitefly species with different efficiencies. For example, the species Mediterranean (MED) in this whitefly species complex transmits tomato yellow leaf curl virus (TYLCV) at a higher efficiency than papaya leaf curl China virus (PaLCuCNV). However, the proteomic responses of whitefly to the infection of different begomoviruses remain largely unknown. METHODS: We used iTRAQ-based proteomics coupled with RT-qPCR to investigate and compare responses of the MED whitefly to the infection of TYLCV and PaLCuCNV. RESULTS: Totally, 259, 395 and 74 differently expressed proteins (DEPs) were identified in the comparisons of TYLCV-infected vs. un-infected, PaLCuCNV-infected vs. un-infected, and TYLCV-infected vs. PaLCuCNV-infected whiteflies, respectively. These proteins appear associated with catabolic process, metabolic process, transport, defense response, cell cycle, and receptor. The comparisons of TYLCV-infected vs. un-infected and PaLCuCNV-infected vs. un-infected shared some similar DEPs, indicating possible involvement of laminin subunit alpha, dystroglycan, integrin alpha-PS2 and cuticle proteins in viral transport as well as the role of putative defense proteins 3 and PITH in anti-viral response. However, 20S proteasome subunits associated with regulation of virus degradation and accumulation were up-regulated in PaLCuCNV-infected but not in TYLCV-infected whiteflies, which may be related to the constraints of PaLCuCNV accumulation in MED. CONCLUSIONS: These findings provide valuable clues for unravelling the roles of some whitefly proteins in begomovirus transmission.