Biological and Molecular Detection of Cucumber mosaic virus (CMV) Isolates Obtained from Izmir

Authors : Sachin Sharma, Mustafa Gümüş

Pages : 199-210

Doi:10.29278/azd.1318370

View : 38 | Download : 49

Publication Date : 2023-12-31

Article Type : Research Paper

Abstract :Objective: This study was intended to determine cucumber mosaic virus (CMV) isolates present in tomato-growing areas of the Izmir province of Turkey in 2021-2022 based on mechanical inoculations and RT-PCR method. Materials and Methods: 17 CMV suspected plant samples previously obtained from Izmir between 2019-2022 and preserved under the appropriate temperature in the Faculty of Agriculture, Department of Plant Protection of Ege University were used to carry out mechanical inoculation of the virus into a number of different test plants consisting of Nicotiana glutinosa, Solanum lycopersicum ‘SC-2121’, and Cucumis sativus ‘Beit Alpha’ cultivars. Virus-inoculated plants were visually evaluated for symptom development, followed by a DAS-ELISA test with CMV-specific antibodies. CMV-positive tobaccos were used to repeat mechanical inoculation of the virus into newly grown test plants, followed by a second ELISA and final RT-PCR test. Results: As a result, CMV-D and CMV-B2 isolates obtained from Izmir produced CMV-like symptoms in the test plants. However, only CMV-D inoculated test plants consistently came out to be positive in the final ELISA and RT-PCR test. CMV-D isolate in tobacco and cucumber induced systemic mosaic and in tomato, caused mosaic, stunting and bushy appearance during 3 weeks of virus inoculation. CMV-D inoculated plants when tested for RT-PCR produced an amplified cDNA band of 280 bp in agarose gel indicating the presence of the virus. Conclusion: This study concludes that CMV causes a variety of symptoms depending upon the viral strain involved, infected host species, and other factors. The presence of CMV-D isolate has been biologically and molecularly identified based on repeated mechanical inoculations in its host species and a final RT-PCR test performed under a controlled experimental setup. Further study of the responsible isolate can be achieved by its genome sequencing and phylogenetic analysis to better understand the viral strain involved in the infection.
Keywords : Hıyar mozaik virüs, mekanik inokulasyon, DAS ELISA, RT PCR