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首页> 中文期刊> 《华北农学报》 >三七NAC转录因子基因PnNAC1的克隆及表达特性分析

三七NAC转录因子基因PnNAC1的克隆及表达特性分析

         
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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.%To reveal the regulatory mechanism of defense response in Panax notoginseng (Burk) FH Chen against diseases,a NAC transcription factor gene was cloned from P.notoginseng,and its expression pattern was analyzed in the present study.One year old P.notoginseng seedlings were used as materials.According to an Unigene from P.notoginseng transcriptome which encoded a NAC transcription factor,the gene-specific primers were designed.Using the rapid amplification of cDNA end technology,a new NAC transcription factor gene of P.notoginseng was obtained which was named PnNAC1.The cDNA sequence of PnNAC1 was 815 bp in length and contained an intact open reading frame (ORF) of 576 bp,a 24 bp 5′-untranslated region (UTR),and a 215 bp 3′-UTR.The ORF of PnNAC1 encoded a putative protein with 191 amino acid residues which had a conserved NAC domain.Real-time PCR results indicated that pre-treatment with methyl jasmonate to roots of P.notoginseng seedlings increased the transcription level of PnNAC1 in roots,then the transcription level of PnNAC1 was further risen after inoculation with Fusarium solani,and its expression level was maximal at 4 h post inoculation with F.solani.For the control (pretreatment with sterile water),PnNAC1 also rapidly responded to infection of F.solani,but its expression level was evidently lower than that in samples which were pre-treated with methyl jasmonate.Moreover,the expression of PnNAC1 was inhibited in the leaves after inoculation with Alternaria panax.Furthermore,the transcription level of PnNAC1 was induced by treatment with four kinds of signaling molecules including methyl jasmonate,ethylene,salicylic acid,and hydrogen peroxid.The above results suggest that PnNAC1 responds to several stress-related signaling molecules and is involved in the defense response of P.notoginseng against the infection of F.solani and A.panax.

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