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花生斑驳病毒印度分离株的分子特征及利用细菌表达的核心衣壳蛋白进行免疫诊断

Molecular characterization of Indian isolate of peanut mottle virus and immunodiagnosis using bacterial expressed core capsid protein.

作者信息

Soumya K, Yogita M, Prasanthi Y, Anitha K, Kishor P B Kavi, Jain R K, Mandal Bikash

机构信息

National Bureau of Plant Genetic Resources, Hyderabad, India.

Division of Plant Pathology, Advanced Centre for Plant Virology, Indian Agricultural Research Institute, New Delhi, 110012 India.

出版信息

Virusdisease. 2014;25(3):331-7. doi: 10.1007/s13337-014-0210-3. Epub 2014 May 8.

Abstract

Peanut mottle virus (PeMoV), a seed borne potyvirus was recorded in India in 1978, however the virus was not characterized at molecular level. In the present study, an isolate of PeMoV infecting peanut in southern India was characterized based on host reactions and coat protein (CP) gene sequence, which revealed that the Indian isolate was very close to a peanut isolate reported from Israel and distinct from pea isolate reported from USA. The core region of CP gene that contained majority of the predicted epitopes was successfully expressed (1.75 mg/l) in Escherichia coli as a 22 kDa protein. A high titer polyclonal antibody (PAb) to the expressed core CP was produced, which efficiently detected PeMoV. The antiserum was useful in specific detection of PeMoV as it showed negligible cross reactivity with the other potyviruses e.g., peanut stripe virus, potato virus Y, papaya ringspot virus and onion yellow dwarf virus. The PAb was validated in ELISA using 1,169 field and greenhouse samples of peanut which showed 1.85-26.3 % incidence of PeMoV in peanut seed multiplication field during 2011-2012. This is the first report of immunodiagnosis of PeMoV with a PAb to recombinant core CP of PeMoV.

摘要

花生斑驳病毒(PeMoV)是一种种传马铃薯Y病毒,1978年在印度被发现,但该病毒尚未在分子水平上得到鉴定。在本研究中,基于寄主反应和外壳蛋白(CP)基因序列对印度南部一株感染花生的PeMoV分离株进行了鉴定,结果表明该印度分离株与以色列报道的一株花生分离株非常接近,与美国报道的豌豆分离株不同。CP基因的核心区域包含了大部分预测的表位,该区域在大肠杆菌中成功表达为一种22 kDa的蛋白(表达量为1.75 mg/l)。制备了针对表达的核心CP的高效价多克隆抗体(PAb),该抗体能有效检测PeMoV。该抗血清可用于PeMoV的特异性检测,因为它与其他马铃薯Y病毒如花生条纹病毒、马铃薯Y病毒、番木瓜环斑病毒和洋葱黄矮病毒的交叉反应可忽略不计。使用1169份花生田间和温室样品通过酶联免疫吸附测定(ELISA)对该PAb进行了验证,结果显示在2011 - 2012年花生种子繁殖田中PeMoV的发病率为1.85% - 26.3%。这是关于用针对PeMoV重组核心CP的PAb对PeMoV进行免疫诊断的首次报道。

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