| 水稻矮缩病毒Pns6和P8蛋白在病毒侵染水稻原生质体后的表达 |
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| 引用本文： | 张洁,＃ 宛柏杰,＃ 尚鹏祥 丁新伦 杜振国 吴祖建.水稻矮缩病毒Pns6和P8蛋白在病毒侵染水稻原生质体后的表达[J].中国水稻科学,2019,33(2):118-123. |
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| 作者姓名： | 张洁 ＃ 宛柏杰 ＃ 尚鹏祥 丁新伦 杜振国 吴祖建 |
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| 作者单位： | 1.福建农林大学 植物病毒研究所/福建省植物病毒学重点实验室/闽台作物有害生物生态防控国家重点实验室, 福州350002;2.江苏沿海地区农业科学研究所, 江苏 盐城 224000 |
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| 基金项目： | 国家自然科学基金资助项目(31672005)；福建农林大学科技创新专项(CXZX2017324, CXZX2016132)。 |
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| 摘 要： | 【目的】为明确水稻矮缩病毒(Rice dwarf virus, RDV) Pns6和P8蛋白在病毒侵染水稻原生质体后的表达动态,【方法】利用PEG介导的病毒侵染原生质体体系,通过免疫荧光和电镜技术分析Pns6、P8蛋白以及病毒粒体在水稻原生质体中的定位;同时,通过Western blot和实时荧光定量PCR分析Pns6和P8蛋白及其RNA在水稻原生质体中的积累量。【结果】病毒接种水稻原生质体48 h后,Pns6蛋白在细胞质中可以形成类似于病毒原质(viroplasm)的点状内含体,P8蛋白也大量的表达。同时,病毒粒体在水稻原生质体中也形成内含体状的结构。病毒接种水稻原生质体12 h后,均可检测到Pns6和P8蛋白的表达,并且在36 h后达到最高值。病毒接种水稻原生质体后,Pns6 RNA在24 h时表达量达到最高,P8 RNA在36 h时表达量达到最高。【结论】RDV侵染水稻原生质体后,Pns6和P8蛋白均有表达,并且病毒也可能通过形成病毒原质来完成病毒在寄主细胞的复制和装配。
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| 关 键 词： | 水稻矮缩病毒 水稻原生质体 Pns6蛋白 P8蛋白 |
| 收稿时间： | 2018-09-29 |
| 修稿时间： | 2018-11-12 |
Expression of Pns6 and P8 Proteins of Rice Dwarf Virus After Virus Infecting Rice Protoplasts |
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| ZHANG Jie,＃,WAN Baijie,＃,SHANG Pengxiang,DING Xinlun,DU Zhenguo,WU Zujian.Expression of Pns6 and P8 Proteins of Rice Dwarf Virus After Virus Infecting Rice Protoplasts[J].Chinese Journal of Rice Science,2019,33(2):118-123. |
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| Authors: | ZHANG Jie ＃ WAN Baijie ＃ SHANG Pengxiang DING Xinlun DU Zhenguo WU Zujian |
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| Institution: | 1.Institute of Plant Virology, Fujian Agriculture and Forestry University / Key Laboratory of Plant Virology of Fujian Province / State Key Laboratory for Ecological Pest Control of Fujian and Taiwan Crops, Fuzhou 350002, China;2.Institute of Agricultural Sciences in the Coastal District of Jiangsu Province, Yancheng 224000, China |
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| Abstract: | 【Objective】In order to clarify expression dynamics of Pns6 and P8 proteins of Rice dwarf virus after virus infecting rice protoplasts, 【Method】subcellular localization of Pns6 and P8 proteins as well as viral particles in rice protoplasts were conducted with a PEG mediated virus infecting protoplast system by immunofluorescence and electron microscope. Meanwhile, accumulation of Pns6 and P8 proteins and their RNAs in rice protoplasts were measured by Western blot and real-time fluorescence quantitative PCR. 【Result】Pns6 protein could form viroplasm-like punctate inclusion bodies in cytoplasm 48 h after virus inoculation, and P8 protein was also expressed in large quantities. Meanwhile, viral particles could form inclusion body-like structure in rice protoplasts. Expression of Pns6 and P8 proteins was both detected at 12 h after virus inoculation, and reached the maximum level after 36 h. At the same time, the expression level of Pns6 RNA peaked at 24 h and that of P8 RNA at 36 h after virus inoculation. 【Conclusion】Pns6 and P8 proteins were both expressed after virus infection, and Rice dwarf virus may also formed viroplasm-like inclusion bodies to complete virus replication and assembly in host cells. |
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| Keywords: | Rice dwarf virus rice protoplast Pns6 protein P8 protein |
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