Department of Plant Systems Biology, VIB Life Sciences Research Institute Gent, Belgium.
Front Plant Sci. 2012 May 24;3:97. doi: 10.3389/fpls.2012.00097. eCollection 2012.
Coordinated, subcellular trafficking of proteins is one of the fundamental properties of the multicellular eukaryotic organisms. Trafficking involves a large diversity of compartments, pathways, cargo molecules, and vesicle-sorting events. It is also crucial in regulating the localization and, thus, the activity of various proteins, but the process is still poorly genetically defined in plants. In the past, forward genetics screens had been used to determine the function of genes by searching for a specific morphological phenotype in the organism population in which mutations had been induced chemically or by irradiation. Unfortunately, these straightforward genetic screens turned out to be limited in identifying new regulators of intracellular protein transport, because mutations affecting essential trafficking pathways often lead to lethality. In addition, the use of these approaches has been restricted by functional redundancy among trafficking regulators. Screens for mutants that rely on the observation of changes in the cellular localization or dynamics of fluorescent subcellular markers enable, at least partially, to circumvent these issues. Hence, such image-based screens provide the possibility to identify either alleles with weak effects or components of the subcellular trafficking machinery that have no strong impact on the plant growth.
蛋白质的协调的、亚细胞内运输是多细胞真核生物的基本特性之一。运输涉及到大量不同的隔室、途径、货物分子和囊泡分拣事件。它在调节各种蛋白质的定位和功能方面也很重要，但该过程在植物中仍然在遗传上定义不足。过去，通过正向遗传学筛选，通过在化学或辐射诱导突变的生物体群体中寻找特定的形态表型，来确定基因的功能。不幸的是，这些简单的遗传筛选在识别新的细胞内蛋白质运输调节剂方面受到限制，因为影响关键运输途径的突变往往导致致死。此外，这些方法的使用受到运输调节剂之间功能冗余的限制。依赖于观察荧光亚细胞标记物的细胞定位或动力学变化的突变体筛选至少部分地规避了这些问题。因此，这种基于图像的筛选提供了识别弱效等位基因或亚细胞运输机制的可能性，这些机制对植物生长没有强烈的影响。
