Comparison of biological and molecular characterization of Iranian lettuce mosaic virus isolates.
Lettuce mosaic virus (LMV) is one of the most damaging viruses in lettuce and endive cultivating regions. In order to review the characteristics of different LMV isolates of Iran during 2004-2005 samples were collected from lettuce fields in Esfahan, Ghom, Khorasan, Khuzestan and Tehran provinces. All of the isolates were detected by LMV polyclonal antiserum (AS-0155, DSMZ Germany) in ELISA and TIPA tests. Biological purification was done for the LMV isolates and then they were maintained and propagated on Chenopodium quinoa. A range of plant species such as C. amaranticolor, C. album, Carthamus tinctorius, Gazania sp., Gomphrena globosa, Pisum sativum, Spinacia oleracea were inoculated with these isolates using potassium phosphate buffer (0/05M). Molecular weight of coat protein was determined by Polyacrylamid gel electrophoresis (PAGE). Immunocapture reverse transcription polymerase chain reaction (IC-RT-PCR) was performed using LMV polyclonal antiserum and specific primer pairs of LMV as described by Zerbini et al. (1995). The amplified fragments were included the whole CP and 3'UTR regions and the nucleotide sequences of them determined. All isolates induced chlorotic local lesions on C. amaranticolor and chlorotic local lesions with symptoms of systemic infection (vein clearing) on C. album. Tehran isolate in addition, caused local lesions on Gomphrena globosa with red border and white centre. This isolate infected Pisum sativum without any symptoms. Back inoculation on C. quinoa and DAS-ELISA confirmed the latent infection. None of these isolates infected Carthamus tinctorius, Gazania sp. and Spinacia oleracea. The molecular weight of coat protein was determined 30.33 kDa. Western-blot proved this band as the coat protein of the virus. IC-RT-PCR amplification of LMV isolates produced the expected size IC-RT-PCR product of 1300 bps. The comparison of nucleotide sequences showed that there were 98% identities.
Ormaz
B
B
Department of Plant Pathology, Faculty of Agriculture, Tehran University, Karaj, Iran.
Winter
S
S
Koohi-Habibi
M
M
Mosahebi
Gh
G
Izadpanah
K
K
D036022:Capsid Proteins / Q000235:genetics / Q000302:isolation & purification; D004797:Enzyme-Linked Immunosorbent Assay; D007492:Iran; D018545:Lactuca / Q000821:virology*; D009029:Mosaic Viruses / Q000145:classification / Q000235:genetics / Q000302:isolation & purification*; D010802:Phylogeny; D010935:Plant Diseases / Q000821:virology*; D018515:Plant Leaves / Q000821:virology; D016133:Polymerase Chain Reaction
D036022:Capsid Proteins