Members of the family Closteroviridae have emerged as a major problem in agricultural crops in the past two decades. Diodia vein chlorosis virus (DVCV) is an understudied whitefly-transmitted closterovirus. Given the presence of the primary host for the virus in major agricultural production areas in the United States, we characterized the virus at the molecular level, demonstrating that it belongs in the genus Crinivirus, developed detection protocols, evaluated its host range among hosts known to harbor viruses closely related to DVCV, and confirmed transmission by a second whitefly species, Trialeurodes vaporariorum.