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| 猕猴桃植株中柑橘叶斑驳病毒实时荧光定量PCR检测技术的建立及应用 | |
| 引用本文： | 刘欢,米伟丽,刘斐,吴薇,吴宽,吴云锋.猕猴桃植株中柑橘叶斑驳病毒实时荧光定量PCR检测技术的建立及应用[J].植物病理学报,2019,49(2):167-173. |
| 作者姓名： | 刘欢 米伟丽 刘斐 吴薇 吴宽 吴云锋 |
| 作者单位： | 旱区作物逆境生物学国家重点实验室,农业部西北黄土高原作物有害生物综合治理重点实验室,西北农林科技大学植物保护学院,杨凌712100; 咸阳市农业科学研究院,咸阳 712000; 西北农林科技大学生命科学学院,杨凌 712100; 杨凌职业技术学院,杨凌 712100 |
| 基金项目： | 高等学校学科创新引智计划项目(B07049); 杨凌职业技术学院科学研究基金计划项目(A2017028); 陕西省重点研发计划(2018NY-103); 陕西省重点研发计划(2018ZDXM-NY-058) |
| 摘 要： | 柑橘叶斑驳病毒(Citrus leaf blotch virus, CLBV)在陕西省栽培猕猴桃中发生普遍。为监测CLBV发生情况,本研究建立了CLBV的实时荧光定量PCR(Real-time fluorescent quantitative polymerase chain reaction,RT-qPCR)检测方法。该方法特异性强,可准确检测目的病毒,标准曲线斜率为-3.378,决定系数R2=0.997 9,扩增效率为97.7%,比普通RT-PCR灵敏度高100倍,可用于猕猴桃植株CLBV的批量检测或低丰度病毒样本(如猕猴桃休眠枝条)的检测。为苗木携带CLBV病毒的早期诊断、果园病毒病预测预报和防控奠定了基础。 |
| 关 键 词： | 猕猴桃 柑橘叶斑驳病毒 实时荧光定量PCR |
| 收稿时间： | 2018-06-13 |
Development and evaluation of a Real-time fluorescent quantitative PCR assay for detection of Citrus leaf blotch virus in kiwifruit plants |
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| LIU Huan,MI Wei-li,LIU Fei,WU Wei,WU Kuan,WU Yun-feng.Development and evaluation of a Real-time fluorescent quantitative PCR assay for detection of Citrus leaf blotch virus in kiwifruit plants[J].Acta Phytopathologica Sinica,2019,49(2):167-173. | |
| Authors: | LIU Huan MI Wei-li LIU Fei WU Wei WU Kuan WU Yun-feng |
| Institution: | State Key Laboratory of Crop Stress Biology for Arid Areas, Key Laboratory of Crop Pest Integrated Pest Management on Crop in Northwestern Loess Plateau, Ministry of Agriculture ,College of Plant Protection, Northwest A＆F University, Yangling 712100, China; Xianyang Agricultural Research Academy, Xianyang 712000, China; College of Life Sciences, Northwest A&F University, Yangling 712100, China; Yangling Technical and Vocational College, Yangling 712100, China |
| Abstract: | During the virus detection of kiwifruit cultivars in Shaanxi province, Citrus leaf blotch virus was found to occur at high frequencies. In order to detect CLBV accurately, a specific RT-qPCR assay based on SYBR Green I fluorescent dye was established. The results showed that the RT-qPCR assay had good specificity and sensitivity for CLBV detection. The slope of the standard curve and determination coefficient R2 were -3.378 and 0.997 9, and the amplification efficiency reached 97.7%, which was 100 times more sensitive than conventional RT-PCR. This method was suited to batch inspection of field samples and low-abundance virus samples (such as dormant shoots of kiwifruit). The RT-qPCR assay laid the foundation for the early diagnosis of CLBV-infected seedlings, forecast and control of CLBV in orchard. |
| Keywords: | kiwifruit Citrus leaf blotch virus RT-qPCR |
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