Biological and molecular detection of Hibiscus chlorotic ringspot virus infecting Hibiscus rosa-sinensis in Iran

During 2011, ten leaf samples of Hibiscus rosa-sinensis with virus-like symptoms including diffuse mottling, numerous chlorotic spots and chlorotic ring spots, were collected from Guilan province, North Iran. The results of mechanical inoculation of sap from these leaves onto different indicator hosts showed local lesions (chlorotic or necrotic) on Gompherena globosa, Chenopodium amaranticolor, C. quinoa and Gossypium hirsutum, while Hibiscus cannabinus (kenaf) showed systemic chlorotic ring spot symptoms. Purified preparations of the isolated virus contained isometric particles approximately 28-30 nm in diameter. A specific band of about 1.3 kb was amplified from all symptomatic leaves using Hibiscus chlorotic ringspot virus (HCRSV) specific primers. Comparative sequence analysis revealed that the IRN. HCRSV (JX865593) isolate shared the greatest nucleotide sequence identity (96.5%) with NC-003608 (Singapore) and the least nt sequence identity (91.5%) with DQ392986 (Taiwan) isolates. The phylogenetic tree showed at least two subgroups for HCRSV isolates in which the Iranian isolate was grouped with Singapore HCSRV (NC-003608). This is the first report of HCRSV in H. rosa-sinensis in Iran.