Poster: Emerging Technologies

Abs # 888: Genomic approaches to auxin biology: a high throughput method for the quantitative analysis of indole-3-acetic acid

Presenter:	Barkawi, Lana S., barka003@umn.edu
Authors	Barkawi, Lana S. (A)   Tam, Yuen Yee  (B)   Cohen, Jerry D (A)   Normanly, Jennifer  (B)
Affiliations:	(A): Department of Horticultural Science, University of Minnesota (B): Department of Biochemistry and Molecular Biology, University of Massachusetts

In order to investigate novel pathways involved in auxin biosynthesis, transport, metabolism and response, we have developed a high throughput screen for indole-3-acetic acid (IAA) levels and synthesis routes. This method will be used as a technique to screen mutant Arabidopsis lines to identify those with altered levels of IAA. These lines will in turn be subjected to a more thorough metabolite profiling. Historically, the quantitative analysis of IAA has been a cumbersome and time-consuming process that does not lend itself to the screening of large numbers of samples. The method described here can be performed with or without a robotic system and involves purification by solid phase extraction and an affinity column in a 96 well format allowing the analysis of 96 samples per day. We show results of experiments designed to test the capacity and robustness of the method, and discuss our progress on expanding this system to include screening for other indolic compounds. Finally, we will show the application of this technique to screen for mutants altered in their ability to produce IAA from tryptophan or via tryptophan independent biosynthesis pathways. Supported by NSF Plant Genome Program grant DBI0077769.