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。
Tomato yellow leaf curl virus (TYLCV) is currently considered as one of the most devastating viruses in cultivated 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.
[1] 蔡健和, 秦碧霞, 朱桂宁, 黄福新, 陈永惠, 李焜华. 番茄黄化曲叶病毒病在广西爆发的原因和防治策略. 中国蔬菜, 2006, (7): 47-48.
[2] 何自福, 虞皓, 毛明杰, 罗方芳, 林奕韩, 王穗涛. 中国台湾番茄曲叶病毒侵染引起广东番茄黄化曲叶病. 农业生物技术学报, 2007, 15(1): 119-123.
[3] 赵统敏, 余文贵, 周益军, 季英华. 江苏省番茄黄化曲叶病毒病(TYLCD)的发生与诊断初报. 江苏农业学报, 2007, 23(6): 654-655.
[4] 吴永汉, 张纯胄, 许方程, 李芳芳, 陈为康, 卢启强, 夏万青. 温州地区番茄曲叶病毒病发生与防治. 中国蔬菜, 2007, (5): 57-58.
[5] 于力, 朱龙英, 万延慧, 杨少军, 张辉, 朱为民. 上海地区番茄黄化曲叶病毒病的鉴定及嫁接接种法研究. 基因组学与应用生物学, 2009, 28(1): 115-118.
[6] Zhang H, Gong HR, Zhou XP. Molecular characterization and pathogenicity of tomato yellow leaf curl virus in China. Virus Gen, 2009, 39(2): 249-255.
[7] 李常保, 柴敏, 李季, 郑建秋. 北京番茄黄化曲叶病毒病的发生及分子检测. 中国蔬菜, 2010, (1): 28-30.
[8] Morilla G, Janssen D, García-Andrés S, Moriones E, Cuadrado IM, Bejarano ER. Pepper (Capsicum an-nuum) is a dead-end host for tomato yellow leaf curl virus. Phytopathology, 2005, 95(9): 1089-1097.
[9] 叶剑, 青玲, 周雪平. 与中国番茄黄化曲叶病毒伴随的DNAβ分子βC1缺失突变体介导的交叉保护作用初步研究. 浙江大学学报 (农业与生命科学版), 2006, 32(5): 479-482.
[10] Del Sal G, Manfioletti G, Schneider C. The CTAB-DNA precipitation method: a common mini-scale preparation of template DNA from phagemids, phages or plasmids suitable for sequencing. Biotechniques, 1989, 7(5): 514-520.
[11] 王伟伟, 朱长青, 刘小花, 陈昆松, 徐昌杰. 番茄叶片基因组DNA快速制备技术及其在基于实时荧光定量PCR的转基因检测中的应用. 遗传, 2011, 33(9): 1017-1022.