Automated Microscopy in Forward Genetic Screening of Arabidopsis - Plant Cell Morphogenesis: Methods and Protocols - page 52

Chapter 4

Automated Microscopy in Forward Genetic Screening
of Arabidopsis

Tereza Dobisová and Jan Hejátko

Abstract

Tightly controlled spatiotemporal specifi city of gene expression is intrinsic to developmental and adapta-
tion responses of living systems throughout the kingdoms. Forward genetic screens employing well-
characterized reporter lines can be used to identify as yet unknown genetic factors driving specifi c aspects
of individual regulatory pathways. However, such screens are demanding with respect to data acquisition
and analysis from thousands of mutant lines. Here, we describe a method that allows screening of a muta-
genized GUS reporter line in Arabidopsis using an automated microscopy imaging system as a tool for
rapid and effi cient identifi cation of mutants with modifi ed expression profi le for a gene of interest.

Key words Automated microscopy, Imaging, Forward genetics, Screening, GUS staining, .slide,
Arabidopsis

1 Introduction

Most genes isolated to date have been identifi ed through diverse
forward genetic screens based upon identifi cation of morphologi-
cal or physiological malfunctions (e.g., refs. [ 1 - 3 ]). The tremen-
dous progress in deciphering the molecular mechanism underlying
particular responses permits screening for more subtle changes that
are specifi c to particular developmental and adaptation pathways.
Furthermore, many if not all of the developmental processes
throughout the kingdoms are associated with the tightly regulated
expression of regulatory genes. Consequently, many of the isolated
mutants are affected in the spatiotemporal specifi city as to the
expression of important developmental regulators (e.g., refs. [ 4 - 7 ]).
Thus, forward genetic screening that specifi cally allows to identify
factors controlling the expression of important developmental reg-
ulators is of great interest.