Abstract

This protocol describes a fluorescence microscope-based screening of Arabidopsis seedlings and describes how to map recessive mutations that alter the subcellular distribution of a specific tagged fluorescent marker in the secretory pathway. Arabidopsis is a powerful biological model for genetic studies because of its genome size, generation time, and conservation of molecular mechanisms among kingdoms. The array genotyping as an approach to map the mutation in alternative to the traditional method based on molecular markers is advantageous because it is relatively faster and may allow the mapping of several mutants in a really short time frame. This method allows the identification of proteins that can influence the integrity of any organelle in plants. Here, as an example, we propose a screen to map genes important for the integrity of the endoplasmic reticulum (ER). Our approach, however, can be easily extended to other plant cell organelles (for example see(1,2)), and thus represents an important step toward understanding the molecular basis governing other subcellular structures.

Links

Publisher Full Text

Authors

Stefano G, Renna L, Brandizzi F

Institution

DOE Plant Research Laboratory, Michigan State University.

Source

Journal of visualized experiments : JoVE :62 2012 pg

MeSH

Arabidopsis
Chromosome Mapping
Endoplasmic Reticulum
Microscopy, Fluorescence
Organelles
Plant Proteins

Pub Type(s)

Journal Article
Research Support, U.S. Gov't, Non-P.H.S.
Video-Audio Media

Language

eng

PubMed ID

22526030