Abstract
Proanthocyanidins (PAs) are colorless flavonoid polymers and deposit in Arabidopsis seed coat specifically. Oxidation of PAs gives rise to brown color of mature seeds. PA accumulation can be affected by a number of growth conditions, such as temperature and sun light. PAs, which are converted from anthocyanidins, can protect seeds from outer environment and have a positive effect in seed longevity (Debeaujon, 2003). Vanillin turns red upon binding to leucoanthoanthocyanidins, catechins and monomers and terminal subunits of PAs (Butler et al., 1982; Deshpande et al., 1986). Based on this principle, PA deposition in Arabidopsis seed coat can be visualized.
Keywords: Seed coat color, Vanillin assay, Arabidopsis, Proanthocyanidins
Materials and Reagents
Equipment
Procedure
Representative data
Figure 1. Comparison of the darkness of PA staining in seed coats among various test transparent mutants. Scale bars = 100 µm. (Wang et al., 2014) Figure 2. Comparison of Arabidopsis siliques between different developmental stages. Stage 1: Young siliques: less than or about one week after pollination. 10 min is enough for reaction between PAs and vanillin. Stage 2: Immature siliques: 1-3 weeks after pollination. 20 to 40 min are required for the reaction. Stage 3: Mature silique: 3 weeks or more after pollination, color of siliques appears pale yellow. 1 h is necessary for the reaction.
Notes
Recipes
Acknowledgments
The work was sponsored by the Natural Science Foundation of China (Grant nos. 31171463). Our protocol is an improvement based on a method described by Hagerman (2002) with sorghum grains.
References
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