Biology Reference
In-Depth Information
Chapter 17
Morphological Analysis of Cell Growth Mutants
in
Physcomitrella
Jeffrey P. Bibeau and Luis Vidali
Abstract
This protocol describes a quantitative analysis of the morphology of small plants from the moss
Physcomitrella patens
. The protocol can be used for the analysis of growth phenotypes produced by tran-
sient RNA interference or for the analysis of stable mutant plants. Information is presented to guide the
investigator in the choice of vectors and basic conditions to perform transient RNA interference in moss.
Detailed directions and examples for fl uorescence image acquisition of small regenerating moss plants are
provided. Instructions for the use of an ImageJ-based macro for quantitative morphological analysis of
these plants are also provided.
Key words
Transient transformation, RNAi,
Physcomitrella patens
, Morphological analysis, Solidity,
Convex hull, Tip growth, Cell polarization, ImageJ, Plant cell growth, Cytoskeleton, Protonema
1
Introduction
Plants develop the great diversity of structures necessary for their
adaption and survival by the coordination of cell expansion and cell
division. Hence, in order to understand plant development, we
need to investigate the molecular mechanisms that control cell
expansion and division, as well as how they are integrated. One of
the most useful tools to investigate these mechanisms is the analy-
sis of phenotypic changes caused by mutations. This is a particu-
larly powerful technique when a quantitative method can be used
to estimate the changes in phenotype. The simple development
program of the moss
Physcomitrella patens
and its ease for genetic
manipulation provide an ideal system to investigate the control of
plant cell expansion and division [
1
,
2
]. We have implemented a
robust and simple method to quantitatively evaluate changes in the
growing pattern of moss plants undergoing early development.
This method can be combined with transient transformation of
RNA interference (RNAi) constructs to investigate the participa-
tion of single genes or gene families in plant cell growth, or
Search WWH ::
Custom Search