A Seed-GUS-Expression Enhancer-trap Library for Germination Research

摘要

Enhancer-trap lines are used to identify tissue- and stage-specific gene expression. An Ambidopsis (Ambidopsis thdiana (L.) Heynh.) enhancer-trap population from the Arabidopsis Biological Resource Center (ABRC), Ohio, USA, has been screened for beta-glucuronidase (GUS) expression in germinating seeds. Application of these enhancer-trap lines for functional analysis of seed germination-associated genes has led to the successful identification of a GATA zinc finger transcription factor crucial for seed germination. A Seed-GUS-Expression enhancer-trap library was donated to the ABRC (stock no. CS24362-CS24480), and is now available to the international seed research community. This library can be used for seed germination research by combining northern blotting and reverse transcription polymerase chain reaction (RT-PCR) expression analysis with functional analysis using knockout plants. Here, an overview of the procedures for utilizing the enhancer-trap library for germination research including some potential pitfalls and hints for troubleshooting experiments is presented.