Identification of Begomoviruses from Three Cryptic Species of Bemisia tabaci (Gennadius) (Hemiptera: Aleyrodidae) in Nepal

The Bemisia tabaci species complex consists of at least 44 cryptic species, which are potential vectors of approximately 320 begomovirus species, most of which are significant plant viruses. However, the relationship of begomovirus transmission through vectors at the cryptic species level is uncertain. In our previous study, three cryptic species (Asia I, Asia II 1, and Asia II 5) of B. tabaci were identified from 76 B. tabaci samples collected across 23 districts in Nepal. Using the same individuals we identified seven different begomovirus species (Squash leaf curl China virus [SLCCNV], Tomato leaf curl New Delhi virus [ToLCNDV], Okra enation leaf curl virus [OELCuV], Synedrella leaf curl virus [SyLCV], Tomato leaf curl Kerala virus [ToLCKeV], Ageratum enation virus [AEV], and Tomato leaf curl Karnataka virus [ToLCKV]) by PCR using universal begomovirus primers. The begomoviruses were detected in 55.26% of whitefly samples, and SLCCNV was the most prevalent species (27.63%). Among the three cryptic species of B. tabaci, the virus detection rate was highest in Asia I (60%), followed by Asia II 1 (58.82%) and Asia II 5 (53.06%). Most viruses were detected in all three species, but AEV and ToLCKV were found only in Asia I and Asia II 1, respectively. Geographic analysis showed that SLCCNV was distributed in the whole country, which is similar to the distribution of the Asia II 5 species, but OELCuV and SyLCV were detected only in the middle region of Nepal. Our results provide important information on the begomovirus profile in Nepal which can be beneficial for plant virus risk assessment and develop the management strategies to reduce the damage of whitefly transmitted viruses.

Implication of the Whitefly Protein Vps Twenty Associated 1 (Vta1) in the Transmission of Cotton Leaf Curl Multan Virus

Cotton leaf curl Multan virus (CLCuMuV) is one of the major casual agents of cotton leaf curl disease. Previous studies show that two indigenous whitefly species of the Bemisia tabaci complex, Asia II 1 and Asia II 7, are able to transmit CLCuMuV, but the molecular mechanisms underlying the transmission are poorly known. In this study, we attempted to identify the whitefly proteins involved in CLCuMuV transmission. First, using a yeast two-hybrid system, we identified 54 candidate proteins of Asia II 1 that putatively can interact with the coat protein of CLCuMuV. Second, we examined interactions between the CLCuMuV coat protein and several whitefly proteins, including vacuolar protein sorting-associated protein (Vps) twenty associated 1 (Vta1). Third, using RNA interference, we found that Vta1 positively regulated CLCuMuV acquisition and transmission by the Asia II 1 whitefly. In addition, we showed that the interaction between the CLCuMuV coat protein and Vta1 from the whitefly Middle East-Asia Minor (MEAM1), a poor vector of CLCuMuV, was much weaker than that between Asia II 1 Vta1 and the CLCuMuV coat protein. Silencing of Vta1 in MEAM1 did not affect the quantity of CLCuMuV acquired by the whitefly. Taken together, our results suggest that Vta1 may play an important role in the transmission of CLCuMuV by the whitefly.

Diversity and evolution of the endosymbionts of Bemisia tabaci in China

The whitefly Bemisia tabaci (Gennadius) (Hemiptera: Aleyrodidae) is a cryptic species complex, including members that are pests of global importance. This study presents a screening of B. tabaci species in China for infection by the primary endosymbiont, Portiera aleyrodidarum, and two secondary endosymbionts, Arsenophonus and Cardinium. The results showed that P. aleyrodidarum was detected in all B. tabaci individuals, while Arsenophonus was abundant in indigenous species of B. tabaci Asia II 1, Asia II 3, and China 1 but absent in the invasive species, Middle East-Asia Minor 1 (MEAM1); Cardinium presented in the Mediterranean (MED), Asia II 1 and Asia II 3 species but was rarely detected in the MEAM1 and China 1 species. Moreover, phylogenetic analyses revealed that the P. aleyrodidarum and mitochondrial cytochrome oxidase 1 (mtCO1) phylograms were similar and corresponding with the five distinct cryptic species clades to some extent, probably indicating an ancient infection followed by vertical transmission and subsequent co-evolutionary diversification. In contrast, the phylogenetic trees of Arsenophonus and Cardinium were incongruent with the mtCO1 phylogram, potentially indicating horizontal transmission in B. tabaci cryptic species complex. Taken together, our study showed the distinct infection status of endosymbionts in invasive and indigenous whiteflies; we also most likely indicated the co-evolution of primary endosymbiont and its host as well as the potential horizontal transfer of secondary endosymbionts.