Washington State University, Department of Plant Pathology, Irrigated Agriculture Research and Extension Center, Prosser, WA, 99350, United States.
Washington State University, Department of Plant Pathology, Irrigated Agriculture Research and Extension Center, Prosser, WA, 99350, United States.
Virus Res. 2021 Apr 15;296:198330. doi: 10.1016/j.virusres.2021.198330. Epub 2021 Feb 5.
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.
樱桃锈斑驳-associated 病毒（CRMaV），属于β-柔曲病毒科 Robigovirus 属，与甜樱桃（Prunus avium）的樱桃锈斑驳病密切相关。在这里，我们报告了一个基于农杆菌的克隆 CRMaV cDNA 构建体接种系统的成功开发。对无病毒樱桃砧木“Krymsk6”[P. cerasus x (P. cerasus x P. maackii)]进行农杆菌接种后，系统叶片在接种后 50 天开始出现黄化斑驳症状。通过 RT-PCR 和 ELISA 证实了“Krymsk6”农杆菌接种植物中存在 CRMaV。随后，从农杆菌接种的“Krymsk6”樱桃中分离出的 CRMaV 通过嫁接可传递到无病毒甜樱桃砧木 P. avium 'Mazzard'上，表现为从嫁接后 35 天开始出现典型的樱桃锈斑驳症状，并通过 Western blot 和 RT-PCR 进一步证实。这些结果明确表明 CRMaV 是甜樱桃锈斑驳病的病原体。本研究中提出的反向遗传学系统可用于研究 CRMaV 的分子生物学，也可作为其他 Robigovirus 属病毒的感染性克隆开发模板。
