三七NAC转录因子基因PnNAC1的克隆及表达特性分析

发布时间：2019-04-27 02:38

【摘要】：为了揭示三七抗病防卫反应的调控机制,对三七的一个NAC转录因子的全长cDNA进行了克隆和表达特性分析。以一年生三七幼苗为材料,根据编码NAC转录因子的三七转录组Unigene设计引物,采用快速扩增cDNA末端技术,克隆得到一个新的NAC转录因子基因,命名为PnNAC1。序列分析表明,该基因全长815 bp,含有24 bp的5'非翻译区和215 bp的3'非翻译区,以及576 bp开放阅读框,共编码191个氨基酸,具有保守的NAC结构域。实时荧光定量PCR结果显示茉莉酸甲酯预处理三七根部大幅提高了PnNAC1在根中的转录水平,接种茄腐镰刀菌后,PnNAC1的表达进一步上升,在接种后4 h转录水平达最大值;无菌水预处理的三七中,PnNAC1也快速响应茄腐镰刀菌的侵染,但表达水平明显低于茉莉酸甲酯预处理的样品。接种人参链格孢后PnNAC1在叶片的表达量受抑制。茉莉酸甲酯、乙烯利、水杨酸和过氧化氢4种信号分子处理三七根部均诱导PnNAC1的表达。表明PnNAC1响应几种逆境相关信号分子,并参与三七对茄腐镰刀菌和人参链格孢的防卫反应。
[Abstract]:In order to reveal the regulatory mechanism of resistance and defense response of Panax notoginseng, the full-length cDNA of a NAC transcription factor of Panax notoginseng was cloned and analyzed. Using one-year-old Panax notoginseng seedlings as materials, primers were designed according to the transcription group Unigene encoding NAC transcription factor, and a novel NAC transcription factor gene named PnNAC1. was cloned by rapid amplification of cDNA terminal technique. Sequence analysis showed that 815 bp, contained a 5 'untranslated region of 24 bp and a 3' untranslated region of 215 bp, as well as an open reading frame of 576 bp, encoding a total of 191 amino acids and a conserved NAC domain. The results of real-time fluorescence quantitative PCR showed that methyl jasmonate pretreatment significantly increased the transcription level of PnNAC1 in roots of Panax notoginseng. After inoculation with Fusarium solanacearum, the expression of PnNAC1 increased further, and reached the maximum at 4 h after inoculation. In aseptic water pretreated Panax notoginseng, PnNAC1 also responded rapidly to Fusarium solanacearum infection, but the expression level was significantly lower than that of methyl jasmonate pretreatment. The expression of PnNAC1 in leaves was inhibited after inoculation with Radix Ginseng. The expression of PnNAC1 was induced by methyl jasmonate, ethephon, salicylic acid and hydrogen peroxide in roots of Panax notoginseng. It is suggested that PnNAC1 responds to several stress-related signal molecules and participates in the defense response of Panax notoginseng to Fusarium solanacearum and Radix Ginseng.
【作者单位】：昆明理工大学生命科学与技术学院;
【基金】：国家自然科学基金项目(81560610) 云南省应用基础研究计划项目(2014FA003)
【分类号】：S567.236

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