摘要
RNA干扰被认为是转录后基因沉默的一种机制。RNA干扰通过小干扰RNA特异性降解目标mRNA来沉默基因表达。本文以烟草花叶病毒126kD蛋白为靶蛋白,在原生质体水平上研究了小干扰RNA对病毒侵染的干扰和抑制作用。ELISA和Northern杂交的实验结果表明,共转染小干扰RNA和TMV的原生质体内检测到较低的病毒含量。在枯斑寄主上,叶片接种小干扰RNA和TMV共转染原生质体后,与对照叶片相比,仅有很少量的病斑产生。这说明,小干扰RNA能够在原生质体水平对病毒起到干扰和抑制作用。因此认为,烟草原生质体系统有利于快速和定量分析小干扰RNA介导对植物病毒的抑制作用。
RNA interference （RNAi） is a mechanism of post-transcriptional gene silencing （PTGS） that has been described in plants. RNAi silences gene expression through small interfering RNA （siRNA） that guides mRNA degradation in a sequence-specific manner. Here, we report that siRNA inhibits virus accumulation by targeting TMV 126 kD protein gene in tobacco protoplasts. ELISA and Northern blot assays displayed that protoplasts electroporated with siRNA ＋ TMV showed a significant reduction of TMV RNA concentration. In hypersensitive host, leaves inoculated by protoplast transfected with siRNA ＋ TMV displayed a few local lesions compared with the controls. These indicated that siRNA could interfere with TMV infection in tobacco protoplasts. Therefore, this protoplast system could facilitate rapid and quantitative analysis for siRNA-mediated interference on plant virus infection.
出处
《植物病理学报》
CAS
CSCD
北大核心
2007年第3期284-288,共5页
Acta Phytopathologica Sinica
基金
This study was supported by grants from the Ministry of Science and Technology of China (100C26216101344)
Thescientific research and development project of Yangtze University(2006)
