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| 番茄黄化曲叶病毒的实时荧光定量PCR 检测 | |
| 引用本文： | 林铭,胡鸿,杜永臣,高建昌,王孝宣,国艳梅. 番茄黄化曲叶病毒的实时荧光定量PCR 检测[J]. 中国蔬菜, 2013, 1(2): 20-26 |
| 作者姓名： | 林铭 胡鸿 杜永臣 高建昌 王孝宣 国艳梅 |
| 作者单位： | 中国农业科学院蔬菜花卉研究所，北京 100081 |
| 基金项目： | 公益性行业(农业)科研专项经费项目(201003065);“948”项目(2011-Z3);农业部园艺作物生物学与种质创制重点实验室资助项目 |
| 摘 要： | 番茄黄化曲叶病毒病（Tomato yellow leaf curl virus disease，TY LCVD）是目前为害番茄生 产的重要病害，准确检测TYLCV 含量是深入研究该病毒致病机理以及抗病育种的基础。根据TYLCV 的 DNA 保守序列设计引物，基于SYBR Green I 检测模式，构建了TYLCV 实时荧光定量PCR 检测体系，并 且对接种病毒30 d 后的感病番茄植株中的TYLCV 进行检测。结果显示，1ng·μL-1 感病番茄总DNA 中 约含有3.47×106 个病毒拷贝。利用实时荧光定量PCR 法比普通PCR 法的灵敏度提高100 倍。 |
| 关 键 词： | 番茄 番茄黄化曲叶病毒 实时荧光定量PCR |
| 收稿时间： | 2012-08-29; |
Real-time PCR for Quantification of Tomato yellow leaf curl virus in Tomato Plants |
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| LIN Ming,HU Hong,DU Yong-chen,GAO Jian-chang,WANG Xiao-xuan,GUO Yan-mei. Real-time PCR for Quantification of Tomato yellow leaf curl virus in Tomato Plants[J]. China Vegetables, 2013, 1(2): 20-26 | |
| Authors: | LIN Ming HU Hong DU Yong-chen GAO Jian-chang WANG Xiao-xuan GUO Yan-mei |
| Affiliation: | Institute of Vegetables and Flowers， Chinese Academy of Agricultural Sciences， Beijing 100081， China |
| Abstract: | Tomato yellow leaf curl virus disease（TYLCVD）is one of the most devastating plant diseases in the world．Accurate quantification of TYLCV is the foundation for further studying the pathogenesis of this virus and also for resistance breeding．This paper constructed a real-time PCR testing system to quantify TYLCV in tomato by the primer，designed on the conservative sequences of TYLCV complete genome，and a testing mode based on SYBR Green I technology．The TYLCV were detected in the infected tomatoes after 30 days post-inoculation，about 3.47×106 virus copies could be detected in 1 ng·μL-1 total DNA in the infected tomato plants．This method is 100-fold more sensitive than the traditional PCR. |
| Keywords: | TYLCV Real-time PCR |
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