番茄黄化曲叶病毒快速分子检测.pdfVIP

HEREDITAS (Beijing) 2012 3 , 34(3): 366―370 ISSN 0253-9772 技术与方法 DOI: 10.3724/SP.J.1005.2012.00366 番茄黄化曲叶病毒的快速分子检测 1 1 1 2 李常保 , 崔彦玲 , 张丽英 , 李传友 1. , 100097; 2. , 100101 番茄黄化曲叶病毒是当前世界范围内危害番茄生产的毁灭性病害。文章针对番茄黄化曲叶病毒全基因组 序列的特异区段自主设计了1 对特异性PCR 引物(上游引物TYLCV-F ：5′-ACGCATGCCTCTAATCCAGTGTA-3′, 下游引物TYLCV-R ：5′-CCAATAAGGCGTAAGCGTGTAGAC-3′), 依据PCR 扩增特异片段543 bp 的有无可以 快速、准确、高效、特异地检测出是否感染了TYLCV 病毒, 这项技术可以方便地应用到工厂化育苗的带毒性 检测、蔬菜大规模生产中植株发病情况的快速检测以及抗病毒育种, 从而为蔬菜安全可持续生产提供科技支撑。 番茄黄化曲叶病毒; 特异引物; 分子检测 Molecular detection of tomato yellow leaf curl virus (TYLCV) 1 1 1 2 LI Chang-Bao , CUI Yan-Ling , ZHANG Li-Ying , LI Chuan-You 1. Beijing Vegetable Research Center, Beijing Academy of Agriculture and Forestry Science, Beijing 100097, China; 2. Institute of Genetics and Developmental Biology, Chinese Academy of Sciences, Beijing 100101, China Abstract: Tomato yellow leaf curl virus (TYLCV) is currently considered as one of the most devastating viruses in culti- vated tomatoes (Solanum lycopersicum) worldwide. We reported here the development of a PCR-based method to quickly detect TYLCV using the primer pairs (TYLCV-F: 5-ACG CAT GCC TCT AAT CCA GTG TA-3 and TYLCV-R: 5-CCA ATA AGG CGT AAG CGT GTA GAC-3), which was designed based on the genome sequence of TYLCV. A TYLCV-specific band of 543 bp was amplified from infected tomato plants. This protocol provides a rapid, reliable, and sensitive tool for molecular detection and identification of TYLCV in the industrial seedling and virus resistance breeding to facilitate safe and sustainable production of tomato. Keywords: tomato yellow leaf curl virus (TYLCV); specific primer; molecular detection