| 水稻黑条矮缩病毒检测体系的优化及田间寄主检测 |
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| 引用本文： | 阴筱,朱芹芹,许斐斐,刘志强,张广民,李向东.水稻黑条矮缩病毒检测体系的优化及田间寄主检测[J].植物病理学报,2013,43(6):647-650. |
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| 作者姓名： | 阴筱 朱芹芹 许斐斐 刘志强 张广民 李向东 |
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| 作者单位： | 山东农业大学植物保护学院植物病毒研究室,泰安 271018 |
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| 基金项目： | 山东省科技攻关项目(2009GG10009021)；山东省现代农业产业技术体系 |
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| 摘 要： | 玉米粗缩病20世纪50年代和90年代中后期在我国部分地区严重发生，2008年至今，该病在黄淮海地区又呈暴发趋势［1］。引起我国北方玉米粗缩病的主要是水稻黑条矮缩病毒（Rice black streaked dwarf virus，RBSDV）［2］。田间寄主植物和传毒昆虫灰飞虱（Laodelphax striatellus Fallen）的发生数量、带毒率与玉米粗缩病的发生密切相关［1］。因此，建立快速灵敏的RBSDV检测体系，明确RBSDV的田间寄主，对有效控制玉米粗缩病具有重要意义。
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| 收稿时间： | 2012-12-06 |
Optimization of RT-PCR detection system for rice black-streaked dwarf virus and detection of its natural hosts |
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| YIN Xiao,ZHU Qin-qin,XU Fei-fei,LIU Zhi-qiang,ZHANG Guang-min,LI Xiang-dong.Optimization of RT-PCR detection system for rice black-streaked dwarf virus and detection of its natural hosts[J].Acta Phytopathologica Sinica,2013,43(6):647-650. |
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| Authors: | YIN Xiao ZHU Qin-qin XU Fei-fei LIU Zhi-qiang ZHANG Guang-min LI Xiang-dong |
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| Institution: | Laboratory of Plant Virology, Department of Plant Pathology, College of Plant Protection, Shandong Agricultural University, Tai’an 271018, China |
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| Abstract: | To establish and optimize the RT-PCR detection system of rice black-streaked dwarf virus (RBSDV), and identify the natural host of RBSDV in the field, four pairs of primers were designed according to the nucleotide sequences of RBSDV S10. RT-PCR detection system for RBSDV was optimized after comparing pri\|mer pair, concentration of Mg2+ and Taq polymerase, and times of RNA dilution. The fourth pair of primers (F4 and R4) was proved to be the most sensitive one. The optimal RT-PCR system was 10× PCR buffer 2.5 μL, 25 mmol/L MgCl2 1.5 μL, dNTP (each 2.5 mmol/L) 2.0 μL, primers F4 and R4 (10 μmol/L) each 1.0 μL, 5 U/ μL Taq DNA polymerse 0.2 μL, template RNA 5.0 μL and sterilized water 11.8 μL to make a total volume of 25 μL. With the method established, RBSDV could be detected from RNA extracted from single Laodelphax striatellus or 30 ng infected maize leaves. Besides plants of family Gramineae, Sonchus brachyotus and Eclipta prostrate from family Asteraceae, and Amaranthus retroflexus from Amaranthaceae were also natural hosts of RBSDV. |
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| Keywords: | Rice black-streaked dwarf virus(RBSDV) detection RT-PCR natural host |
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