A vector system for fast-forward studies of the HOPZ-ACTIVATED RESISTANCE1 (ZAR1) resistosome in the model plant Nicotiana benthamiana
A vector system for fast-forward studies of the HOPZ-ACTIVATED RESISTANCE1 (ZAR1) resistosome in the model plant Nicotiana benthamiana
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DOI：
10.1093/plphys/kiab471
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发表时间：
2021-10-11
期刊：
影响因子：
7.4
通讯作者：
Adachi, Hiroaki
中科院分区：
文献类型：
--
作者：
Harant, Adeline;Pai, Hsuan;Adachi, Hiroaki
本烟已成为拟南芥的一个补充实验系统。它主要通过瞬时基因表达实现体内快进分析，在植物免疫研究中特别受欢迎。最近，随着拟南芥HopZ激活的RSISTANCE1(ZAR1)抗性小体的发现，我们对核苷酸结合的富含亮氨酸重复序列(NLR)的植物免疫受体的了解有了很大的进步。在这里，我们描述了一个由72个质粒组成的载体系统，该系统能够对本底夜蛾中的ZAR1抗性小体进行功能研究。我们证明了ZAR1在卷绕类NLRs(CC-NLRs)中脱颖而出，因为它在远缘双子叶植物中高度保守，并确认NbZAR1是拟南芥ZAR1的N.benthamiana同源基因。效应器激活和自动激活的NbZAR1在N.benthamiana中触发细胞死亡反应，这一活动依赖于一个功能性的N-末端α1螺旋。C-末端标记的NbZAR1在N.benthamiana中仍然具有功能，因此能够在这个植物系统中进行细胞生物学和生化研究。我们的结论是，NbZAR1开源PZA质粒收集为拟南芥在植物抗性小体研究中形成了一个额外的实验系统。
Nicotiana benthamiana has emerged as a complementary experimental system to Arabidopsis thaliana. It enables fast-forward in vivo analyses primarily through transient gene expression and is particularly popular in the study of plant immunity. Recently, our understanding of nucleotide-binding leucine-rich repeat (NLR) plant immune receptors has greatly advanced following the discovery of the Arabidopsis HOPZ-ACTIVATED RESISTANCE1 (ZAR1) resistosome. Here, we describe a vector system of 72 plasmids that enables functional studies of the ZAR1 resistosome in N. benthamiana. We showed that ZAR1 stands out among the coiled coil class of NLRs (CC-NLRs) for being highly conserved across distantly related dicot plant species and confirmed NbZAR1 as the N. benthamiana ortholog of Arabidopsis ZAR1. Effector-activated and autoactive NbZAR1 triggers the cell death response in N. benthamiana and this activity is dependent on a functional N-terminal alpha 1 helix. C-terminally tagged NbZAR1 remains functional in N. benthamiana, thus enabling cell biology and biochemical studies in this plant system. We conclude that the NbZAR1 open source pZA plasmid collection forms an additional experimental system to Arabidopsis for in planta resistosome studies.