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克隆烟草NtNAC-R1基因,并进行瞬时表达分析,探讨其是否含有内含子及其在烟碱生物合成中的调控作用。以烟草基因组DNA为模板克隆得到NAC转录因子NtNAC-R1,鉴定结果表明,该基因含有2个内含子。通过构建真核表达载体,建立瞬时表达分析体系,RT-PCR检测结果显示NtNAC-R1基因过表达,烟碱生物合成关键基因PMT表达量升高,JA信号途径标记基因PDF1.2和IAA响应基因IAA13表达量降低。瞬时表达分析表明,该基因受JA信号和IAA信号途径的交互对话影响,进而调控烟碱的生物合成。
Cloning of tobacco NtNAC-R1 gene, and transient expression analysis to explore whether it contains introns and its regulatory role in nicotine biosynthesis. The NAC transcription factor NtNAC-R1 was cloned by using tobacco genomic DNA as a template, and the result showed that the gene contained two introns. RT-PCR results showed that NtNAC-R1 gene overexpression and nicotinic biosynthesis key gene PMT expression increased, JA signal pathway marker gene PDF1.2 and IAA response gene IAA13 expression decreased. Transient expression analysis showed that this gene was affected by the interactive dialogue between JA signal and IAA signaling pathway, which further regulated nicotine biosynthesis.