拟南芥NADPH氧化酶RBOHD羧基端倒数第三位氨基酸在其介导活性氧迸发中的重要作用（英文）

论文摘要

目的:解析呼吸爆发氧化酶同系物蛋白D（RBOHD）介导活性氧迸发的分子机制。创新点:首次研究RBOHD蛋白羧基端在植物体内活性氧迸发中的作用,并对其机制进行初步探究。方法:本文利用正向遗传学方法筛选得到在多种病原物相关分子模式（PAMP）处理后活性氧不迸发的突变体delt。然后结合图位克隆和全基因测序技术,发现DELT1编码了RBOHD蛋白。DELT1-2在RBOHD羧基端倒数第三位谷氨酸位置发生了突变。深入分析发现,谷氨酸的突变不影响DELT1-2表达、蛋白定位和互作等功能,但会导致植物不响应PAMP诱导的气孔关闭。结论:RBOHD羧基端倒数第三位谷氨酸对其功能发挥起着决定作用。

论文目录

• 1 Introduction
• 2 Materials and methods
•   2.1 Plant materials and growth conditions
•   2.2 Chemiluminescence assay of ROS production
•   2.3 Map-based cloning of DELT1-1 gene
•   2.4 Whole genome sequencing
•   2.5 RNA isolation and qRT-PCR
•   2.6 Gene cloning and plasmid construction
•   2.7 Agrobacterium tumefaciens-mediated transi-ent protein expression in Nicotiana benthamiana
•   2.8 SLC assay
•   2.9 BiFC analysis
•   2.1 0 Protein extraction and immunoblot analysis
•   2.1 1 Stomatal closure assay
• 3 Results
•   3.1 Isolation of delt mutants
•   3.2 Mapping of delt1 mutant
•   3.3 delt1-1 and delt1-2 mutations are allelic to rbohD null mutations
•   3.4 E919K mutation does not affect RBOHD tran-script and protein levels
•   3.5 E919K mutation does not affect RBOHD subcellular localization
•   3.6 E919K mutation does not affect RBOHD as-sociation with proteins
•   3.7 E919 of RBOHD is critical for its function in stomatal closure
• 4 Discussion
• 5 Conclusions
• Contributors
• Compliance with ethics guidelines
• List of electronic supplementary materials

文章来源

类型: 期刊论文

作者: Qiu-ying LI,Ping LI,Nang MYINT PHYU SIN HTWE,Ke-ke SHANGGUAN,Yan LIANG

关键词: 活性氧,氧化酶,微生物相关分子模式,脂多糖,呼吸爆发氧化酶同系物蛋白

来源: Journal of Zhejiang University-Science B（Biomedicine & Biotechnology） 2019年09期

年度: 2019

分类: 工程科技Ⅱ辑,基础科学

专业: 生物学

单位: Ministry of Agriculture Key Lab of Molecular Biology of Crop Pathogens and Insects,College of Agriculture and Biotechnology,Zhejiang University

基金: the National Natural Science Foundation of China(No.31622006),the Postdoctoral Science Foundation of China(Nos.2018M630683 and 2018T110601)

分类号: Q946.5

页码: 713-729

总页数: 17

文件大小: 4031K

下载量: 87