title: Purification and immuno-gold labeling of lily mottle virus from lily leaves.
abstract: 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.
authors.affiliation: Cold and Arid Regions Environmental and Engineering Research Institute, Chinese Academy of Sciences, Lanzhou 730000, China. Electronic address: zyubao@yahoo.com.
authors.forename: Yubao
authors.initials: Y
authors.lastname: Zhang
authors.affiliation: Cold and Arid Regions Environmental and Engineering Research Institute, Chinese Academy of Sciences, Lanzhou 730000, China. Electronic address: w.yajun@yahoo.com.
authors.forename: Yajun
authors.initials: Y
authors.lastname: Wang
authors.affiliation: Cold and Arid Regions Environmental and Engineering Research Institute, Chinese Academy of Sciences, Lanzhou 730000, China. Electronic address: wxhcas@lzb.ac.cn.
authors.forename: Zhongkui
authors.initials: Z
authors.lastname: Xie
authors.affiliation: Cold and Arid Regions Environmental and Engineering Research Institute, Chinese Academy of Sciences, Lanzhou 730000, China. Electronic address: wangruoyu@lzb.ac.cn.
authors.forename: Ruoyu
authors.initials: R
authors.lastname: Wang
authors.affiliation: Cold and Arid Regions Environmental and Engineering Research Institute, Chinese Academy of Sciences, Lanzhou 730000, China. Electronic address: guozhhong@hotmail.com.
authors.forename: Zhihong
authors.initials: Z
authors.lastname: Guo
authors.affiliation: Wageningen University, Department of Plant Sciences, Centre for Crop Systems Analysis, Droevendaalsesteeg 1, 6708PB Wageningen, The Netherlands. Electronic address: wopke.vanderwerf@wur.nl.
authors.forename: Wopke
authors.initials: W
authors.lastname: van der Werf
authors.affiliation: Cold and Arid Regions Environmental and Engineering Research Institute, Chinese Academy of Sciences, Lanzhou 730000, China. Electronic address: lelenmg@163.com.
authors.forename: Le
authors.initials: L
authors.lastname: Wang
mesh_terms: D007150:Immunohistochemistry; D027762:Lilium / Q000821:virology*; D016253:Microscopy, Immunoelectron; D018515:Plant Leaves / Q000821:virology*; D017800:Potyvirus / Q000302:isolation & purification*; D013194:Staining and Labeling; D014773:Virology / Q000379:methods
keywords: Immuno-gold labeling assay; Lily mottle virus; Purification; Transmission electron microscope