Zhang Yubao, Wang Yajun, Xie Zhongkui, Wang Ruoyu, Guo Zhihong, van der Werf Wopke, Wang Le
Cold and Arid Regions Environmental and Engineering Research Institute, Chinese Academy of Sciences, Lanzhou 730000, China.
Wageningen University, Department of Plant Sciences, Centre for Crop Systems Analysis, Droevendaalsesteeg 1, 6708PB Wageningen, The Netherlands.
J Virol Methods. 2016 Jun;232:33-8. doi: 10.1016/j.jviromet.2016.02.001. Epub 2016 Feb 27.
Lily mottle virus (LMoV) is prevalent in Lilium species worldwide causing dwarfing, flower breaking, and reduced bulb yield. In this paper, an easy to use and efficient procedure is described for purification of LMoV from lily leaves. The resulting sample is characterized by a 260/280 nm absorbance ratio of 1.20 at a concentration of 1.27 mg/ml. The procedure results in high protein purity and particle integrity as shown by UV-spectrophotometry, polyacrylamide gel electrophoresis (PAGE), Western blotting, reverse transcriptase (RT)-PCR and transmission electron microscopy (TEM) in combination with immuno-gold labeling. This is the first time that an immuno-gold labeling (IGL) assay was performed to identify a virus of lily. Purified products can be used as a source of antigen in the preparation of antibodies against LMoV and may assist in the development of a diagnostic test for LMoV and in epidemiological surveys.
百合斑驳病毒（LMoV）在全球百合属植物中普遍存在，会导致植株矮化、花朵变色以及种球产量降低。本文描述了一种从百合叶片中纯化LMoV的简便高效方法。所得样品在浓度为1.27 mg/ml时，260/280 nm吸光度比值为1.20。紫外分光光度法、聚丙烯酰胺凝胶电泳（PAGE）、蛋白质免疫印迹法、逆转录酶（RT）-PCR以及结合免疫金标记的透射电子显微镜（TEM）结果表明，该方法可实现高蛋白质纯度和病毒粒子完整性。这是首次采用免疫金标记（IGL）检测法鉴定百合病毒。纯化产物可作为抗原用于制备抗LMoV抗体，并可能有助于开发LMoV诊断检测方法及开展流行病学调查。
