A single amino acid substitution in the movement protein enables the mechanical transmission of a geminivirus.

Begomoviruses of the Geminiviridae are usually transmitted by whiteflies and rarely by mechanical inoculation. We used tomato leaf curl New Delhi virus (ToLCNDV), a bipartite begomovirus, to address this issue. Most ToLCNDV isolates are not mechanically transmissible to their natural hosts. The ToLCNDV-OM isolate, originally identified from a diseased oriental melon plant, is mechanically transmissible, while the ToLCNDV-CB isolate, from a diseased cucumber plant, is not. Genetic swapping and pathological tests were performed to identify the molecular determinants involved in mechanical transmission. Various viral infectious clones were constructed and successfully introduced into Nicotiana benthamiana, oriental melon, and cucumber plants by Agrobacterium-mediated inoculation. Mechanical transmissibility was assessed via direct rub inoculation with sap prepared from infected N. benthamiana. The presence or absence of viral DNA in plants was validated by PCR, Southern blotting, and in situ hybridization. The results reveal that mechanical transmissibility is associated with the movement protein (MP) of viral DNA-B in ToLCNDV-OM. However, the nuclear shuttle protein of DNA-B plays no role in mechanical transmission. Analyses of infectious clones carrying a single amino acid substitution reveal that the glutamate at amino acid position 19 of MP in ToLCNDV-OM is critical for mechanical transmissibility. The substitution of glutamate with glycine at this position in the MP of ToLCNDV-OM abolishes mechanical transmissibility. In contrast, the substitution of glycine with glutamate at the 19th amino acid position in the MP of ToLCNDV-CB enables mechanical transmission. This is the first time that a specific geminiviral movement protein has been identified as a determinant of mechanical transmissibility.

Complete nucleotide sequence of capsicum chlorosis virus isolated from Phalaenopsis orchid and the prediction of the unexplored genetic information of tospoviruses.

Phalaenopsis orchids are popular ornamentals all over the world. A tospovirus, capsicum chlorosis virus (CaCV-Ph) had been identified as the cause of chlorotic ringspots on leaves of Phalaenopsis orchids in Taiwan. The tripartite genome of CaCV-Ph was found to contain 3608, 4848 and 8916 nt of S, M and L RNAs, respectively. Phylogenetic analysis of the nucleocapsid (N) protein confirmed that CaCV-Ph is a member of the watermelon silver mottle virus (WSMoV) serogroup in the genus Tospovirus. Based on the relations among the nonstructural protein (NSs), glycoprotein (GnGc), thrips genera, host and geographical distribution, tospoviruses and thrips could be classified into two major types: WSMoV-Thrips-Asian and Tomato spotted wilt virus (TSWV)-Frankliniella-EuroAmerican. The proline (P(459)) of all tospoviral Gn proteins was indispensable for thrips transmission, but the RGD motif, which is maintained by only six tospoviruses, may not be required for thrips transmission. An RdRp catalytic domain found in the conserved region of the L protein may recognize the typically conserved sequences on the 5' and 3' terminal regions (5' AGAGCAAU 3').