大麦黄矮病毒运动蛋白是RNA沉默抑制因子

doi:10.3724/SP.J.1259.2012.00395

植物学报 ›› 2012, Vol. 47 ›› Issue (4): 395-404.DOI: 10.3724/SP.J.1259.2012.00395

大麦黄矮病毒运动蛋白是RNA沉默抑制因子

申永梅^†, 刘国富^†, 杨纪君, 霍晓辉, 曹雪松^*

聊城大学生命科学学院, 聊城 252059

收稿日期:2011-10-28 修回日期:2012-03-30 出版日期:2012-07-01 发布日期:2012-07-26
通讯作者: 曹雪松
基金资助:
国家自然科学基金项目

The Movement Protein of Barley Yellow Dwarf Virus is a Suppressor of RNA Silencing

Yongmei Shen^†, Guofu Liu^†, Jijun Yang, Xiaohui Huo, Xuesong Cao^*

School of Life Sciences, Liaocheng University, Liaocheng 252059, China

Received:2011-10-28 Revised:2012-03-30 Online:2012-07-01 Published:2012-07-26
Contact: Xuesong Cao

摘要/Abstract

摘要： 大麦黄矮病毒(barley yellow dwarf virus, BYDV)属黄症病毒科家族, 其基因组包含6个开放阅读框(open reading frames, ORFs)。将BYDV的6个基因分别克隆到pWEIMING101载体上, 得到重组基因。电击转化农杆菌后, 利用农杆菌瞬时表达方法渗透注射转GFP基因的本氏烟草16c植株的叶片, 在长波长紫外灯下观察GFP的表达, 并通过Northern blot证明所得现象。研究结果表明, BYDV的PAV株系ORF4编码的运动蛋白(movement protein, MP)是RNA沉默抑制因子, 其表达可以抑制局部和系统RNA沉默。BYDV-MP与GFP的双链RNA(dsGFP)共表达后仍能抑制RNA沉默, 荧光强度与叶片中GFP的mRNA和其沉默降解形成的siRNA的量有对应关系, 其N端核定位序列对抑制局部基因沉默起主要作用, 第5、6位氨基酸是抑制基因沉默的关键氨基酸。BYDV-MP单独渗透注射的部位均产生细胞死亡。

Abstract: The genome of barley yellow dwarf virus (BYDV), a member of the Luteoviridae family, contains 6 open reading frames (ORFs). We cloned the 6 ORFs into plasmid of pWEIMING101 and electroporated them into Agrobacterium tumefaciens for transient coexpression of BYDV ORFs and GFP in Nicotiana benthamiana line 16c, a transgenic GFP plant. Northern blot analysis revealed the movement protein (MP) encoded by BYDV-PAV ORF4 as a suppressor of both local and systemic GFP RNA silencing and RNA silencing triggered by double-stranded GFP RNA. The fluorescent intensity was associated with steady state levels of GFP mRNAs and its degradation product of siRNAs. The nuclear localization signal in the N terminus of the MP and amino acid 5 and 6 were the functional domain and motif, respectively, essential for inhibiting RNA silencing. Transient expression of BYDV-MP alone in infiltrated tissue resulted in cell death.

中图分类号:

申永梅, 刘国富, 杨纪君, 霍晓辉, 曹雪松. 大麦黄矮病毒运动蛋白是RNA沉默抑制因子. 植物学报, 2012, 47(4): 395-404.

Yongmei Shen, Guofu Liu, Jijun Yang, Xiaohui Huo, Xuesong Cao. The Movement Protein of Barley Yellow Dwarf Virus is a Suppressor of RNA Silencing. Chinese Bulletin of Botany, 2012, 47(4): 395-404.

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https://www.chinbullbotany.com/CN/Y2012/V47/I4/395

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大麦黄矮病毒运动蛋白是RNA沉默抑制因子

The Movement Protein of Barley Yellow Dwarf Virus is a Suppressor of RNA Silencing

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