中文摘要：被黄瓜花叶病毒感染的辣椒会造成巨大的经济损失，进而促进了抗黄瓜花叶病毒转基因辣椒的发展。本文利用农杆菌介导法研发了抗病毒转基因辣椒。免疫印迹分析显示，纯化的CMV外壳蛋白（CMV-CP）有25 kDa的分子量。在胶内酶解和淡化后，CMV-CP内部的肽片段通过基质辅助激光解吸电离飞行时间质排成序列。作者用定量酶联免疫吸附法检测抗黄瓜花叶病毒转基因辣椒。本文还提供了对转基因辣椒的表达蛋白进行进一步安全评估的基本信息。研究发现，转基因辣椒在任何耕地范围的不同生长阶段无统计学差异。这项研究的结果有助于对新型抗黄瓜花叶病毒转基因辣椒进行安全性评价，并为与其他抗病毒转基因辣椒的对比提供了一个标准。
外文摘要：The great economic losses caused by Cucumber mosaic virus (CMV) infection of peppers has led to the development of genetically modified (GM) CMV-resistant peppers. We developed virus-resistant pepper plants using Agrobacterium tumefaciens-mediated transformation. The expressed recombinant protein was purified using nickel-nitrilotriacetic acid resin and immunoaffinity chromatography, and purity was assessed by sodium dodecyl sulfate polyacrylamide gel electrophoresis. Immunoblot analysis revealed the purified CMV coat protein (CMV-CP) had a molecular mass of 25 kDa. After in-gel digestion and desalting, the internal peptide fragments of CMV-CP were sequenced by matrix-assisted laser desorption/ionization time of flight. Most GM pepper and Escherichia coli BL21 internal peptides had identical peptide sequences and contained 137 of 183 whole peptides in CMV-CP. A quantitative enzyme-linked immunosorbent assay was performed to detect CMV-resistant GM peppers. We also provide basic information about the expressed protein in GM peppers for further safety assessment. The contents of soluble protein and CMV-CP were measured in GM and control peppers cultivated in three different areas of Korea. Statistical significance in terms of cultivation areas, harvest times, generations, and plant tissue origin were determined based on a P value of 0.05. The highest amount of CMV-CP was detected at the seedling stage from plant grown in each region. T3 and T5 showed significantly different levels of CMV-CP from T4 in leaves in the whorl stage. No statistical differences were observed among GM peppers at different stages of maturity in any cultivation area. The results from this study contribute to the safety evaluation of newly designed CMV-resistant GM peppers and provide a standard against which to compare other virus-resistant GM peppers.
外文关键词：CMV-resistant peppers; soluble protein; enzyme-linked immunosorbent assay (ELISA); safety evaluation
作者：Kim, Eunji; Noh, Hee Min; Phat, Chanvorleak; 等
作者单位：Chung Ang Univ
期刊名称：KOREAN JOURNAL OF HORTICULTURAL SCIENCE & TECHNOLOGY
期刊影响因子：0.261
出版年份：2016
出版刊次：12
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