Single-Embryo RT-PCR Assay to Study Gene Expression Dynamics During Embryogenesis in Arabidopsis thaliana

摘要

We have developed a single-embryo RT-PCR protocol for studying gene expression during plant embryogenesis. Four genes, glyceraldhyde-3-phosphate dehydrogenase (GAPC), shoot-meristemless (STM), monopteros (MP), and shaggy-like kinase etha (ASKη), from Arabidopsis thaliana were used to test the sensitivity and reliability of this method by analyzing the differential signal intensities of their RT-PCR products. The method could detect genes expressed during embryogenesis at a single-embryo level and, therefore, can be used to identify phenotypes. When in vitro, embryogenesis also is used to control the time course of zygote development exactly. The single-embryo RT-PCR protocol becomes a powerful method to survey the dynamics of specific gene expression.