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根据豆梨(Pyrus calleryana Dence)接种黑斑病菌后的转录组测序结果,筛选出一个上调表达的NAC转录因子基因。从豆梨中获得了该基因的全长片段,并将其命名为PcNAC1。该基因的开放阅读框为792 bp,编码263个氨基酸。将PcNAC1蛋白序列与其他物种蛋白序列进行对比,并构建系统进化树后发现,其与小麦TaNAC4及拟南芥ATAF1具有较高同源性。通过实时荧光定量分析PcNAC1在豆梨不同组中的表达发现,PcNAC1在茎、叶和根中表达量较高,在花和果中表达量较低。构建了亚细胞定位载体,瞬时侵染本氏烟,通过激光共聚焦显微镜观察融合蛋白在细胞内的分布情况,发现该基因行使功能的主要区域定位在细胞核。进一步在本氏烟中瞬时表达PcNAC1,接种烟草疫霉病菌后发现PcNAC1可以通过调控植物激素通路防卫反应基因的表达来增强植物的抗病性。
According to the sequencing results of transcriptome of Pyrus calleryana Dence inoculated with Alternaria alternata, an up-regulated NAC transcription factor gene was screened out. The full-length fragment of this gene was obtained from Pear and named PcNAC1. The open reading frame of this gene is 792 bp, encoding 263 amino acids. Comparing the protein sequence of PcNAC1 with that of other species and constructing a phylogenetic tree, we found that it has high homology with wheat TaNAC4 and Arabidopsis ATAF1. The results showed that PcNAC1 was highly expressed in stem, leaf and root, and lower in flowers and fruits. The subcellular localization vector was constructed and transiently infected with N. benthamiana. The distribution of the fusion protein in the cells was observed by laser scanning confocal microscopy. The main region of the function of the gene was located in the nucleus. Further, PcNAC1 was transiently expressed in N. benthamiana. Inoculation with Phytophthora nicotianae showed that PcNAC1 enhanced plant resistance by regulating the expression of plant hormone pathway defense response genes.