Whirly转录因子对非寄主菌诱导水稻HR反应的负调控作用

Negative Regulation of Rice Whirly Transcription Factor for the Hypersensitive Response Induced by Non-host Pathogen Bacterium

用RNA干扰技术研究了Whirly转录因子对水稻非寄主抗性反应HR的调控作用。PCR扩增编码Whirly转录因子的Oswhirly基因片段,分别以正向、反向顺序连接到载体pTCK303的两个多克隆位点中,构建Oswhirly基因沉默的RNA干扰载体pTCK303-Oswhy,经农杆菌EHA105菌株转化水稻细胞,结果发现转基因水稻细胞受非寄主菌诱导后,HR反应明显,细胞死亡率显著高于野生型及空载体转化水稻细胞。Oswhirly基因沉默导致HR反应增强,初步揭示Whirly转录因子可能是调控水稻HR反应的负调控因子。

The regulation function of rice Whirly transcription factor for rice hypersensitive response （HR） induced by non-host pathogen bacterium was studied with RNAi technology. A fragment of Oswhirly gene encoding rice Whirly transcription factor was amplified by PCR method, then inserted into 2 flanking MCSs of pTCK303 vector in forward or reverse direction. The constructed vector pTCK-Oswhy was the RNAi vector for Oswhirly gene silence and was used to transform rice cells mediated by the Agrobacterium tumefaciens EHA105 strain. Results showed that hypersensitive response was obviously induced for pTCK303-Oswhy transformed rice cells after inoculation with non-host pathogen bacterium. The cell death rate of transformed rice cells was significantly higher than that of wild type or empty vector transformed rice cells. These results suggested that the Whirly transcription factor might be a negative regulator for the control of the rice induced hypersensitive response.