%0 Journal Article
%T 小麦条锈菌效应蛋白基因 PSTG_23616的时空表达特征分析<br>Spatial and Temporal Expression Pattern of Effector Protein Gene PSTG_23616 in Puccinia striiformis f.sp.tritici
%A 宋
%A 平
%A 谭成龙
%A 郭
%A 嘉
%A 戚
%A 拓
%A 刘
%A 芃
%A 郭
%A 军
%J 西北农业学报
%D 2016
%R 10.7606/j.issn.1004-1389.2016.09.002
%X 明确条锈菌（Puccinia striiformis f.sp.tritici,Pst）效应蛋白基因 PSTG_23616的时空表达特征，旨在解析该基因在病菌生长发育及致病过程中的作用。利用RT-PCR和PCR克隆获得 PSTG-23616的cDNA和基因组DNA全长；借助生物信息学软件分析PSTG-23616蛋白序列特征并构建系统进化树；通过酵母分泌系统、农杆菌介导的瞬时表达系统、实时荧光定量RT-PCR（qRT-PCR）、亚细胞定位技术初步明确该基因的功能。结果表明， PSTG_23616基因组全长937 bp，开放阅读框（open reading frame，ORF）全长699 bp。生物信息学软件预测结果显示，PSTG-23616信号肽为位于N端的1～22个氨基酸；酵母分泌系统分析表明，PSTG-23616的信号肽具有分泌功能，为典型的分泌蛋白；注射含有 PSTG_23616的重组农杆菌菌液，可抑制由BAX引起的细胞程序性死亡（Programmed cell death，PCD）反应；qRT-PCR结果表明， PSTG_23616在小麦条锈菌初侵染过程中一直上调表达，24 h表达量达到顶峰；亚细胞定位分析表明， PSTG_23616在整个细胞均有分布。推测，该效应蛋白参与条锈菌侵染结构的分化，同时在条锈菌与小麦互作过程中发挥毒性功能。<br>In this study,we characterized spatial and temporal expression pattern of Pst (Puccinia striiformis f.sp. tritici) effector protein PSTG_23616 for further functional characterization of PSTG_23616.The full-length of cDNA and DNA sequences of PSTG_23616 were obtained by RT-PCR and PCR technology.Bioinformatic methods were used to analyze molecular properties of PSTG_23616.The yeast secretion system,agrobacterium-mediated transient expression system,quantitative real-time PCR (qRT-PCR) technique and subcellular localization assays were applied to functional characterization of PSTG_23616.The full-length of genomic DNA of PSTG_23616 was 937 bp,and the open reading frame (ORF) of PSTG_23616 was 699 bp.Sequence analysis indicated that the signal peptide of PSTG_23616 was located in the N terminal 1-22 amino acids.The yeast secretion system assays indicated PSTG_23616 was a typical secreted protein with the signal peptide for protein secretion.The secreted protein PSTG_23616 could inhibit the PCD triggered by apoptosis protein BAX in mice.qRT-PCR assays revealed that PSTG_23616 transcripts was up-regulated during early infection stages of Pst and the maximum expression reached at 24 h.However,the expression of PSTG_23616 was down-regulated gradually after 72 h till to 264 h. Subcellular localization assays revealed that PSTG_23616 was localized throughout the cells of wheat protoplasts or tobacco cells.Our data showed that PSTG_23616 may be involved in differentiation of infection structures of Pst and had a virulence role during Pst-wheat interaction.
%K 小麦条锈菌 效应蛋白 PSTG_23616 实时荧光定量RT-PCR 亚细胞定位&lt
%K br&gt
%U http://xbnyxb.alljournals.cn/ch/reader/view_abstract.aspx?file_no=20160902&flag=1