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作者	单位
陈增菊，王婷，汤春蕾，赵梦鑫，康振生，王晓杰	（西北农林科技大学植物保护学院/旱区作物逆境生物学国家重点实验室，陕西杨凌 712100）

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中文摘要:

为明确小麦条锈菌吸器特异表达效应蛋白Hasp58抑制植物免疫的功能，利用PCR获得Hasp58的全长cDNA序列；借助生物信息学软件预测Hasp58的信号肽；将Hasp58构建到pGR106载体用于农杆菌（Agrobactrium tumefacien）介导的烟草瞬时表达系统，明确Hasp58抑制细胞坏死的毒性功能；将Hasp58构建到pCambia1302载体用于烟草细胞定位，明确Hasp58定位情况；将Hasp58构建到pEDV6载体用于荧光假单胞杆菌（Pseudomonas fluorescens）介导的小麦瞬时表达系统,对小麦胼胝质、过敏性坏死面积、活性氧、菌丝面积、菌丝长度进行统计分析，明确Hasp58抑制寄主植物PAMPs引发的免疫反应（PTI）和效应蛋白诱导的免疫反应（ETI）功能。结果表明，条锈菌Hasp58的cDNA全长为1 026 bp，编码342个氨基酸，N端含26_aa的信号肽；通过农杆菌侵染，在烟草中瞬时表达Hasp58，发现其能够抑制由BAX诱导的细胞坏死，为条锈菌的候选效应蛋白；烟草细胞定位发现，含有信号肽和缺失信号肽的Hasp58与GFP融合表达均定位在细胞质，推测该效应蛋白在胞质作用；利用细菌的三型分泌系统在小麦中瞬时表达Hasp58，发现其能够抑制荧光假单胞杆菌引起的胼胝质积累，导致由无毒性条锈菌小种CYR23引起的小麦过敏性坏死面积和活性氧减少，使菌丝面积和长度增加。推测小麦条锈菌效应蛋白Hasp58可抑制寄主植物的PAMPs引发的免疫反应（PTI）和效应蛋白诱导的免疫反应（ETI），并促进自身侵染。

英文摘要:

The purpose of this study was to clarify the function that Hasp58 inhibits plant immunity. Hasp58 is an effector protein specific expression in haustorium of Puccinia striiformis f. sp. tritici. The full-length cDNA of Hasp58 was obtained by PCR. Prediction of the signal peptide of Hasp58 was conducted by adopting the bioinformatics software. Hasp58 was constructed into pGR106 plasmid for Agrobactrium tumefacien-mediated transient expression system of tobacco,and the toxic function of Hasp58 inhibiting cell death was clarified. Hasp58 was constructed into pCambia1302 plasmid for tobacco cell localization to clarify the localization of Hasp58. Hasp58 was constructed into pEDV6 plasmid for Pseudomonas fluorescens-mediated wheat transient expression system. Statistical analysis was performed on wheat callose,necrotic area,reactive oxygen,hyphal area and hyphal length,to clarify the function of Hasp58 inhibiting PAMPs-induced immune(PTI) and effector-induced immune(ETI) of host plants.The results indicated that the full-length sequence of cDNA of the Hasp58 of Puccinia striiformis f. sp. tritici was 1 026 bp,encoding 342 amino acids with a 26_aa signal peptide at the N-terminal. Hasp58 inhibited BAX-induced cell death in tobacco transiently expressing Hasp58,which was as a candidate effector protein from Puccinia striiformis f. sp. tritici. Hasp58-GFP containing either a signal peptide or a deletion signal peptide were accumulated in the cytoplasm in tobacco,suggesting that Hasp58 played a role in plant cytoplasm. Transiently expressing Hasp58 in wheat via the type Ⅲ secretion system inhibited the accumulation of callose caused by Pseudomonas fluorescens,and reduced the necrotic area and the reactive oxygen caused by the non-toxic race CYR23 of Puccinia striiformis f. sp. tritici，resulting in increasing the hyphal area and hyphal length. Taken together,Hasp58 from Puccinia striiformis f. sp. tritici inhibits PAMPs-triggered immune(PTI) and effector-triggered immune(ETI) of host plants and promotes infection.

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