First Detection of Tomato Infectious Chlorosis Virus (TICV) in Greenhouse Tomato Using RT-PCR and Electron Microscopy

Abstract

Tomato infectious chlorosis virus (TICV), a member of the genus Crinivirus, is an emerging threat to greenhouse tomato production systems due to its impact on plant vigor and yield reduction. The present study reports the first detection and molecular confirmation of TICV from symptomatic greenhouse-grown tomato plants exhibiting interveinal chlorosis, leaf brittleness, and stunted growth. Leaf samples were collected from affected plants and subjected to total RNA extraction followed by RT-PCR amplification using TICV-specific primers targeting conserved genomic regions. The presence of expected amplicons confirmed viral infection, and sequencing analysis validated close similarity with previously reported TICV isolates from other geographic regions. Transmission electron microscopy of infected leaf tissue revealed characteristic flexuous filamentous virions consistent with crinivirus morphology, further supporting molecular findings. Phylogenetic analysis grouped the detected isolate within established TICV clades, indicating genetic conservation across global populations. Field observations suggested potential involvement of whitefly vectors (Trialeurodes vaporariorum) in virus dissemination within greenhouse conditions. Disease incidence was higher under warm and humid microclimatic environments favoring vector multiplication and virus spread. The study highlights the first molecular and ultrastructural confirmation of TICV in greenhouse tomato cultivation, expanding its known geographical distribution. These findings emphasize the urgent need for routine molecular diagnostics and strict vector management strategies to prevent further spread in protected cultivation systems. Integrated disease management approaches, including the use of virus-free planting material, vector exclusion techniques, and regular monitoring, are recommended. Overall, the detection of TICV underscores the increasing importance of crinivirus surveillance in greenhouse horticulture and provides a foundation for developing effective management strategies to safeguard tomato production under intensifying protected agriculture systems.