Abstract
Cherry rusty mottle-associated virus (CRMaV), which belongs the genus Robigovirus of the family Betaflexiviridae, is strongly associated with cherry rusty mottle disease of sweet cherry, Prunus avium. Here, we report on the successful development of an Agrobacterium-based inoculation system for a cloned CRMaV cDNA construct. Agro-inoculation of virus-free cherry rootstock ‘Krymsk6’ [P. cerasus x (P. cerasus x P. maackii)] resulted in the development of chlorotic yellow mottle symptoms on systemic leaves beginning at 50 days post inoculation. The presence of CRMaV in ‘Krymsk6’ agro-inoculated plants was confirmed by RT-PCR and ELISA. Subsequently, CRMaV from agro-inoculated ‘Krymsk6’ was graft-transmissible onto virus-free sweet cherry rootstock P. avium ‘Mazzard’ as evidenced by the production of typical cherry rusty mottle symptoms beginning at 35 days post grafting, and further confirmed by western blotting and RT-PCR. These results showed conclusively that CRMaV is the causal agent of cherry rusty mottle disease in sweet cherry. The reverse genetic system presented in this study can be used as a tool to investigate the molecular biology of CRMaV and also a template for infectious clone development for other viruses in the genus Robigovirus.
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