Engineering a robust infectious clone and gene silencing vector from blackberry yellow vein associated virus

Criniviruses are emerging pathogens responsible for significant disease outbreaks worldwide. Among them, blackberry yellow vein-associated virus (BYVaV) is prevalent in blackberry-producing areas of the United States and, when present in the blackberry yellow vein disease complex with other viruses, can lead to substantial crop losses. To better understand BYVaV biology and its role in virus complex disease development, we developed a BYVaV-derived infectious clone and a virus-induced gene silencing (VIGS) vector. The infectious clone successfully induced systemic infection and symptom development in Nicotiana benthamiana. Additionally, transmission of the recombinant virus to indicator plants was confirmed using the whitefly vector Trialeurodes vaporariorum. The infectious clone was subsequently modified into a VIGS vector, with the foreign insert remaining stable for the length of the study. This work provides essential tools for advancing the study of BYVaV biology and conducting genomic studies in its natural hosts.