CELL RES :万建民院士丨钙离子启动免疫系统的分子机制(附NMT实验体系)
重大进展 | 万建民连发Science及Cell Res,揭示钙离子启动免疫系统的分子机制
- 期刊:cell research
- 主题:钙离子启动免疫系统的分子机制
- 标题:A cyclic nucleotide-gated channel mediates cytoplasmic calcium elevation and disease resistance in rice
- 影响因子:17.848
- 检测指标:Ca2+流速
- 检测部位:水稻叶肉细胞
- Ca2+流速流实验处理方法:水稻幼苗,10uM chitin或10uM flg22肽瞬时胁迫
- Ca2+流速流实验测试液成份:0.2mM CaCl2, 0.1mM NaCl, 0.1mM MgCl2 and 0.1mM KCl,pH 5.2
- 作者:中国农科院万建民、王家昌
英文摘要
The transient elevation of cytoplasmic calcium is essential for pathogen-associated molecular pattern (PAMP)-triggered immunity (PTI). However, the calcium channels responsible for this process have remained unknown.
Here, we show that rice CDS1 (CELL DEATH and SUSCEPTIBLE to BLAST 1) encoding OsCNGC9, a cyclic nucleotide-gated channel protein, positively regulates the resistance to rice blast disease. We show that OsCNGC9 mediates PAMP-induced Ca2+ influx and that this event is critical for PAMPs-triggered ROS burst and induction of PTI-related defense gene expression. We further show that a PTI-related receptor-like cytoplasmic kinase OsRLCK185 physically interacts with and phosphorylates OsCNGC9 to activate its channel activity.
Our results suggest a signaling cascade linking pattern recognition to calcium channel activation, which is required for initiation of PTI and disease resistance in rice.
中文摘要(谷歌机翻)
细胞质钙的瞬时升高对病原体相关分子模式(PAMP)- 触发免疫(PTI)至关重要。然而,负责该过程的钙通道仍然未知。
在这里,我们显示编码OsCNGC9(环核苷酸门控通道蛋白)的水稻CDS1(CELL DEATH和SUSCEPTIBLE to BLAST 1)正向调节对稻瘟病的抗性。我们显示OsCNGC9介导PAMP诱导的Ca2+内流,并且该事件对于PAMPs触发的ROS爆发和诱导PTI相关的防御基因表达是至关重要的。我们进一步显示PTI相关受体样细胞质激酶OsRLCK185与OsCNGC9物理相互作用并使其磷酸化以激活其通道活性。
我们的研究结果表明信号级联将模式识别与钙通道激活联系起来,这是启动水稻PTI和抗病性所必需的。
结果表明:响应于几丁质或flg22刺激,WT叶肉细胞比cds1叶肉细胞表现出强烈且快速的Ca2+流入(图3a,b和补充信息,图S6)。这些结果表明OsCNGC9可以介导水稻PTI中的Ca2+流入,并且这种能力在cds1突变体中受损。
结果表明:响应几丁质刺激,Nipponbare叶肉细胞比Osrlck185/ 55双突变体叶肉细胞,表现出快速的Ca2+内流(图e)。此外,在几丁质处理Oscerk1敲除突变体后未观察到显着的Ca2+流入(图f)。这些结果共同表明OsRLCK185及其紧密同源物OsRLCK55参与水稻抗稻瘟病和PAMP诱导的Ca2+内流的调节。
结果表明:在PAMPs刺激后,与Kitaake植物相比,OsCNGC9-OE转基因植物的叶肉细胞显示出更强的Ca2+流入。结合其他实验表明,OsCNGC9是水稻PTI的限速正调控因子。
文章链接:https://www.nature.com/articles/s41422-019-0219-7