伊兹密尔黄瓜花叶病毒(CMV)分离物的生物学和分子检测

Sachin Sharma, Mustafa Gümüş
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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. 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摘要

研究目的本研究旨在根据机械接种和 RT-PCR 方法确定 2021-2022 年土耳其伊兹密尔省番茄种植区存在的黄瓜花叶病毒(CMV)分离株。 材料与方法:利用之前于 2019-2022 年间从伊兹密尔获得并在埃格大学植物保护系农学院适当温度下保存的 17 个 CMV 疑似植物样本,将病毒机械接种到由 Nicotiana glutinosa、Solanum lycopersicum 'SC-2121' 和 Cucumis sativus 'Beit Alpha' 栽培品种组成的多个不同测试植物中。对接种病毒的植株进行目测症状发展评估,然后用 CMV 特异性抗体进行 DAS-ELISA 检测。用 CMV 阳性烟草重复机械接种病毒到新长出的试验植株中,然后进行第二次 ELISA 和最后的 RT-PCR 测试。 检测结果结果:从伊兹密尔获得的 CMV-D 和 CMV-B2 分离物在测试植物中产生了类似 CMV 的症状。然而,在最终的 ELISA 和 RT-PCR 测试中,只有接种 CMV-D 的测试植物始终呈阳性。烟草和黄瓜中的 CMV-D 分离物会诱发系统性马赛克,番茄中的 CMV-D 分离物会在接种病毒 3 周后引起马赛克、发育不良和灌木状外观。接种 CMV-D 的植株在进行 RT-PCR 检测时,在琼脂糖凝胶中产生了 280 bp 的 cDNA 扩增带,表明病毒的存在。 结论本研究得出结论,CMV 会导致多种症状,这取决于所涉及的病毒株、感染的宿主种类和其他因素。根据对宿主物种的反复机械接种和在受控实验装置下进行的最终 RT-PCR 测试,从生物学和分子学角度确定了 CMV-D 分离物的存在。通过基因组测序和系统发育分析,可对该分离株进行进一步研究,以更好地了解参与感染的病毒株。
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İzmir’den Alınan Hıyar mozaik virüsü (CMV) İzolatlarının Biyolojik ve Moleküler Tespiti
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.
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