Abstract：
The endomembrane system in plants is composed of interconnected membrane organelles that contribute to intracellular structure and function. These organelles include the endoplasmic reticulum (ER), Golgi apparatus, vacuole, trans-Golgi network, and prevacuolar compartment or multivesicular body. Through vesicle-mediated transport, secreted proteins are synthesized in the ER and subsequently transported along the secretory pathway to the vacuole or outside of cells to fulfill specialized functions. Genetic screening is a crucial method for studying plant protein secretion. It entails identifying phenotypic differences resulting from genetic mutations, such as ethyl methanesulfonate, T-DNA insertion, and RNAi, to investigate gene function and discover mutants with specific traits or gene functions. Significant progress has been achieved in the study of plant protein secretion through genetic screening. In this protocol, we provide a step-by-step guide to studying the protein secretion pathway using a genetic screen approach. We use the example of the free 1 suppressor of Arabidopsis thaliana and oil body mutants of Marchantia polymorpha. Additionally, we offer an overview of genetic screening and briefly summarize the emerging technologies in the field of protein secretion research.
摘要：
植物的内膜系统由相互连接的膜细胞器组成，这些细胞器有助于细胞内的结构和功能。这些细胞器包括内质网（ER），高尔基体,液泡,跨高尔基网络，和前液泡室或多泡体。通过囊泡介导的转运，分泌的蛋白质在ER中合成,随后沿着分泌途径转运至液泡或细胞外,以实现特定功能。遗传筛选是研究植物蛋白分泌的重要方法。它需要识别基因突变导致的表型差异，如甲磺酸乙酯，T-DNA插入,RNAi,研究基因功能并发现具有特定性状或基因功能的突变体。通过遗传筛选对植物蛋白分泌的研究取得了重大进展。在这个协议中,我们提供了使用基因筛选方法研究蛋白质分泌途径的分步指南.我们使用拟南芥的游离1抑制剂和Marchantiapolymorpha的油体突变体的例子。此外,我们对基因筛选进行了概述，并简要总结了蛋白质分泌研究领域的新兴技术。
