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Development:北京林业大学丨固醇调控FLS2蛋白胞吞的新机制(钙信号)​(附NMT实验体系)

转自中关村旭月非损伤微测技术产业联盟


期刊:Development
主题:固醇调控FLS2蛋白胞吞的新机制(钙信号)
标题:Sterols regulate endocytic pathways during flg22-induced defense responses in Arabidopsis
影响因子:5.413
检测指标:Ca2+流速
检测样品:拟南芥叶片
Ca2+流速实验处理方法:7日龄拟南芥,0.1 μM flg22处理
Ca2+流实验测试液成份:
推荐测试液:0.1mM CaCl2,pH 6.0
作者:北京林业大学林金星、李晓娟

英文摘要

The plant transmembrane receptor kinase FLAGELLIN SENSING 2 (FLS2) is crucial for innate immunity. Although previous studies have reported FLS2-mediated signal transduction and endocytosis via the clathrin-mediated pathway, whether additional endocytic pathways affect FLS2-mediated defense responses remains unclear.

Here, we show that the Arabidopsis thaliana sterol-deficient mutant steroid methyltransferase 1 displays defects in immune responses induced by the flagellin-derived peptide flg22. Variable-angle total internal reflection fluorescence microscopy (VA-TIRFM) coupled with single-particle tracking showed that the spatiotemporal dynamics of FLS2-GFP changed on a millisecond time scale and that the FLS2-GFP dwell time at the plasma membrane increased in cells treated with a sterol-extracting reagent when compared with untreated counterparts.

We further demonstrate that flg22-induced FLS2 clustering and endocytosis involves the sterol-associated endocytic pathway, which is distinct from the clathrin-mediated pathway. Moreover, flg22 enhanced the colocalization of FLS2-GFP with the membrane microdomain marker Flot 1-mCherry and FLS2 endocytosis via the sterol-associated pathway. This indicates that plants may respond to pathogen attacks by regulating two different endocytic pathways.

Taken together, our results suggest the key role of sterol homeostasis in flg22-induced plant defense responses.

 

中文摘要(谷歌机翻译)

植物跨膜受体激酶FLAGELLIN SENSING 2(FLS2)对于先天免疫至关重要。尽管以前的研究已经报道了通过网格蛋白介导的途径介导的FLS2介导的信号转导和胞吞作用,但尚不清楚其他内吞途径是否会影响FLS2介导的防御反应。

在这里,我们显示拟南芥的甾醇缺乏突变体类固醇甲基转移酶1在鞭毛蛋白衍生的肽flg22诱导的免疫反应中显示缺陷。可变角度全内反射荧光显微镜(VA-TIRFM)结合单粒子跟踪显示,在处理的细胞中,FLS2-GFP的时空动态在毫秒级变化,质膜上的FLS2-GFP停留时间增加与未经处理的对应物相比,含固醇提取剂。

我们进一步证明,flg22诱导的FLS2聚集和内吞涉及与固醇相关的内吞途径,这不同于网格蛋白介导的途径。此外,flg22通过固醇相关途径增强了FLS2-GFP与膜微区标记Flot 1-mCherry和FLS2内吞的共定位。这表明植物可以通过调节两种不同的内吞途径来响应病原体侵袭。

两者合计,我们的结果表明固醇稳态在flg22诱导的植物防御反应中的关键作用。

结果表明:相比野生型,smt1(甾醇甲基转移酶1)突变体表现出迅速且强烈的Ca2+吸收速率高于根冠(R0和R1),低于根伸长区(R2)。增加,说明突变体对于flg22更为敏感。smt1突变没有改变FLS2的同源寡聚状态,但影响FLS2簇形成。smt1突变体中FLS2的内吞功能受到损伤。由上述结果得到一个假设,即甾醇相关的内吞途径对于flg22诱导的FLS2动力学和植物防御至关重要。

 

文章链接:https://dev.biologists.org/content/145/19/dev165688

 

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