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ARS Home » Pacific West Area » Corvallis, Oregon » Forage Seed and Cereal Research Unit » Research » Publications at this Location » Publication #206825

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

item LIU, PO-PO
item Martin, Ruth

Submitted to: Book Chapter
Publication Type: Book / Chapter
Publication Acceptance Date: 8/31/2006
Publication Date: 10/31/2006
Citation: Liu, P., Martin, R.C., Hewitt, J.R., Koizuka, N., Homrichhausen, T.M., Nonogaki, H. 2006. A seed-GUS-expression Enhancer-trap Library for Germination Research. In: Navie, S., Adkins, S. and Ashmore, S., editors. Seeds: Biology, Development and Ecology. CAB International 2006. p. 159-168.

Interpretive Summary: This book chapter is based on a talk given at the 8th International Workshop on Seeds. Genes have promoters which can be thought of as on/off switches which determine where and when a gene is expressed. Enhancer-trap lines are generated by transforming plants with a minimal promoter (MP, which by itself cannot turn a gene on) fused to a reporter gene (a gene that allows you to visualize where and when it is expressed). If the MP–reporter fusion is inserted next to an enhancer, the reporter gene is expressed in an organ-, tissue-, or cell-specific pattern as directed by the enhancer (Campisi et al., 1999). Enhancer-trap lines could be very useful for identifying potentially important factors of seed germination. A brief overview of procedures for utilizing the Seed-Gus-Expression enhancer-trap library that was recently donated to the Arabidopsis Biological Resource Center (ABRC) is presented. This is a library of transformed plants that show GUS expression in seeds which can be utilized to identify factors important in seed germination.

Technical Abstract: Enhancer-trap lines are used to identify tissue- and stage-specific gene expression. An Arabidopsis (Arabidopsis thaliana (L.) Heynh.) enhancer-trap population from the Arabidopsis Biological Resource Center (ABRC), Ohio, USA, has been screened for '-glucuronidase (GUS) expression in germinating seeds. Application of these enhancer-trap lines for func-tional 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 are presented.