Abstract
The goal of this protocol is to assay for defects in synthesis/secretion/release of seed coat mucilage by ruthenium red staining of mature whole seeds.The mucilage secretory cells of the Arabidopsis seed coat synthesize and secrete a large quantity of primarily pectinaceous mucilage to a ring-shaped apical domain during their differentiation. This makes them an excellent model system to identify genes involved in both cell wall synthesis and secretion (et al., 2000). When wild-type seeds are incubated in ruthenium red stain, hydrated mucilage is extruded from epidermal cells and a ‘halo’ of red-stained mucilage is observed surrounding the seed (Western et al., 2000). Reduced mucilage staining may result from defects in cell wall biosynthesis, secretion, or impaired release upon hydration.
Keywords: Arabidopsis, Seed coat, Mucilage, Ruthenium red, Pectin
Materials and Reagents
Equipment
Procedure
Notes
Recipes
Acknowledgments
This protocol was developed by T. Western at the University of British Columbia while in the laboratory of Dr. George Haughn, funded through National Sciences and Engineering Research Council grants to G. Haughn. While the first use was shown in Western et al. (2000) referenced in the text, the protocol was first described in Western et al. (2001) and elaborated for details of EDTA pretreatment in Arsovski et al. (2009a), referenced in the text.
References
If you have any questions/comments about this protocol, you are highly recommended to post here. We will invite the authors of this protocol as well as some of its users to address your questions/comments. To make it easier for them to help you, you are encouraged to post your data including images for the troubleshooting.