玉米ZmNLS1介导的内质网相关的蛋白降解调控植株生长发育的分子机制研究

玉米内质网相关的蛋白降解（ERAD）系统的组分及其在生长发育中的作用尚不清楚。利用植株发育突变体nls1，我们克隆了ZmNLS1，它编码玉米membralin蛋白。前期研究显示，ZmNLS1可能是ERAD系统的组分，其突变诱发了nls1的细胞程序化死亡（PCD）。本研究旨在揭示ZmNLS1在玉米生长发育中的作用及其调控机制，包括：1）构建敲除、抑制和过表达转基因系，检测植株表型，明确其在生长发育中的作用；2）进一步检测ZmNLS1与ERAD组分的互作，及其突变对ERAD能力的影响，明确它是否作为ERAD组分参与蛋白稳态的维持；3）检测nls1中非折叠蛋白反应（UPR）是否被激活，明确ZmNLS1突变是否诱发了持续的内质网胁迫；4）上述结果结合已完成的RNA-seq和表型数据等，解析ZmNLS1突变诱发的UPR信号调控PCD的可能途径，揭示ZmNLS1突变导致植株矮小、叶片显著变窄的本质。

The components of the maize endoplasmic reticulum-associated degradation (ERAD) system and its role in growth and development are unclear. Using the plant development mutant nls1, we cloned ZmNLS1, which encodes the maize membralin protein. Our previous results suggested that ZmNLS1 may be a component of ERAD system and its mutation lead to programmed cell death (PCD) of the mutant nls1. The purpose of this study is to reveal the role and regulatory mechanism of ZmNLS1 in maize growth and development, including: 1) construction of knockout, suppression and overexpression transgenic lines, testing plant phenotypes, and clarifying its role in growth and development; 2) further test the interaction between ZmNLS1 and ERAD components and the effect of its mutation on ERAD ability to determine whether it is involved in the maintenance of protein homeostasis as an ERAD component; 3) determine whether ZmNLS1 mutation causes continuous endoplasmic reticulum stress by detecting whether the unfolded protein response (UPR) is activated in the mutant nls1; 4) combining the above results with the completed RNA-seq and phenotype data to reveal possible pathways for the regulation of PCD by UPR signals caused by ZmNLS1 mutation, revealing the nature of ZmNLS1 mutation leading to short plants and narrow leaves.