Detection of Cucumber Mosaic Virus by Immunestrips and Reverse Transcriptase Polymerase Chain Reaction

Abstract

This study was conducted to detect the cucumber mosaic virus (CMV) on cucumber plants , determine the hostrange and the source of primary infection ,by using indicator plants, immunostrips test and RT- PCR.The infection was done by using the sap. of cucumber plants containing CMV including some indicator plants like cucumber (Cucumis sativus L.),tomato(Lycopersicon esculentum Mill), zucchini ( Cucurbita pepo L), Pumpkin (Cucurbita maxima L.), eggplants (Solanum melongena L.)cowpea (Vigna unguiculata L.) tobacco (Nicotiana tabacum cv. samsun, Nicotiana rustica and Nicotiana tabacum cv. Russian ) Pumpkin (Cucurbita maxima L) Wall goosefoot(Chenopodium murale L.).The results showed that all above-mentioned plants were infected with CMV. The symptoms of infection were mosaic, Malt formation and stunting of cucumber, tomato leaves changed to filiform shape. The symptoms of zucchini, Pumpkin and eggplant in form of systemic mosaic and stunting of eggplants, while the symptoms upon cowpea plants were pale local lesion leaves, in the wall goosefoot plants, the symptoms were unorganized spots. The three varieties of tobacco showed vein clearing . The test of immunostrip showed the existence of CMV in cucumber plants, tomato plants, pumpkin plant and cowpea plants. by clear deposition line compare to control plants.The diagnosis of CMV virus was confirmed by Reverse Transcriptase Polymerase Chain Reaction (RT--PCR). The plant specimens collected from areas understudy were given positive results, as the RT-PCR reaction with primer CMV1 and CMV2 for CMV-coat protein gene (CMV-cp Gene)yielded 486-488 base pair DNA bands while the specimens from non- infected plants amplified no products. These results indicated that the CMV virus detected in present study belong to subgroup I. This study is the first in the region that have been used genetic diagnosis of CMV virus.