Cytological, Histological and Molecular Characterization of Two Isolates of Cucumber Mosaic Virus (CMV) in Egypt

Q4 Agricultural and Biological Sciences
E. Wagih, M. Zalat, M. Kawanna
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引用次数: 2

Abstract

Two isolates of Cucumber mosaic virus (CMV), CMV-wild tobacco (from Alexandria governorate) and CMV-cucumber (from Kafr El-Sheikh governorate) were investigated in this study. Cytological studies on epidermal strips of Nicotiana glutinosa leaves separately infected with each isolate revealed the presence of viral crystalline inclusion bodies within the infected cells. Electron microscopy of ultrathin sections of CMV infected N. glutinosa leaves showed significant alterations in the shape and internal structure of chloroplasts. The cell wall had serrated edges in infected cells but was more severe in cells infected with CMV-wild tobacco isolate compared to those infected with CMV-cucumber isolate. CMV-cucumber isolate was partially purified from systemically infected leaves of N. glutinosa. The ratio A260/ 280 was 1.0 and the concentration of the virus in the preparation was estimated using an extinction coefficient of E260nm0.1%, 1cm = 5. Yield of purified virus was about 2.8 mg/100 g fresh weight of infected N. glutinosa leaves. Electron microscopy of the purified preparation of CMV showed the presence of numerous spherical particles with a mean particle diameter of 28 nm. Amplified real-time reverse transcription-polymerase chain reaction (qRT-PCR) product of coat protein gene of each isolate was purified and sequenced. Sequences of both isolates had been submitted to GenBank Database and ware assigned accession number (LT669766) for CMV-cucumber isolate and (LT706517) for CMV-wild tobacco isolate. The sequences were edited using Chromas Pro. Version 1.34 software and compared with previously subgrouping of 27 isolates of the virus retrieved from the GenBank database. Both CMV-wild tobacco and CMV-cucumber isolates were closely related to the isolate with the accession number AJ585086 with a similarity of 97.07% and 98.54%, respectively, suggesting that the two isolates belong to subgroup II. According to the available literature, this is the first report in Egypt where CMV isolates belonging to subgroup II have been obtained
两株埃及黄瓜花叶病毒(CMV)的细胞学、组织学和分子特性
对黄瓜花叶病毒(CMV)的两个分离株进行了研究,分别是产自亚历山大省的CMV野生烟草和产自Kafr El-Sheikh省的CMV黄瓜。对不同菌株分别感染的烟叶表皮条进行细胞学研究,发现在感染细胞内存在病毒结晶包涵体。对巨细胞病毒感染的玉米叶片进行电镜观察,发现其叶绿体形态和内部结构发生了显著变化。感染细胞的细胞壁呈锯齿状边缘,但野生烟草分离物感染细胞的细胞壁比黄瓜分离物感染细胞的细胞壁更严重。从系统侵染的黄瓜叶片中分离得到部分cmv -黄瓜分离物。A260/ 280的比值为1.0,用消光系数E260nm0.1%, 1cm = 5估计制剂中的病毒浓度。纯化后的病毒产量约为2.8 mg/ 100g鲜重。电镜显示,CMV纯化后的样品中存在大量球形颗粒,平均粒径为28 nm。对各分离物外壳蛋白基因的实时逆转录-聚合酶链反应扩增产物进行纯化和测序。两株分离物的序列均已提交到GenBank数据库,并分别为cmv -黄瓜分离物(LT669766)和cmv -野生烟草分离物(LT706517)分配了登录号。使用Chromas Pro编辑序列。版本1.34软件,并与先前从GenBank数据库检索的27株病毒亚组进行比较。cmv -野生烟草和cmv -黄瓜分离株与加入号AJ585086的分离株亲缘关系密切,相似度分别为97.07%和98.54%,属于II亚群。根据现有文献,这是在埃及首次获得属于II亚群的巨细胞病毒分离株的报告
本文章由计算机程序翻译,如有差异,请以英文原文为准。
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来源期刊
International Journal of Phytopathology
International Journal of Phytopathology Agricultural and Biological Sciences-Agricultural and Biological Sciences (miscellaneous)
CiteScore
1.20
自引率
0.00%
发文量
11
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