Abstract
This assay analyzes Arabidopsis seed germination in response to gibberellic acid (GA). During seed imbibition, visible physiological changes allow precise determination of germination rate. This protocol utilizes a stereoscopic microscope to improve characterization of seed germination process.
Background
Seed germination is a critical process of the plant life cycle controlled by phytohormones, such as GA and abscisic acid (ABA), and environmental factors. Seed germination comprises two physiological processes, including seed coat (testa) and endosperm ruptures. Usually, penetration of endosperm by the radicle indicates that germination is complete. Previous studies generally use endosperm rupture to calculate germination rate. However, seed coat rupture also measures the progression of seed germination. This protocol utilizes a stereoscopic microscope to provide a visible and precise calculation of seed germination, including the rates of both seed coat and endosperm rupture (Zhong et al., 2015).
Materials and Reagents
Equipment
Software
Procedure
Data analysis
Using Photoshop CS5 software, count the number of the seeds with a ruptured testa or endosperm at each time point. The germination rate is indicated by the percentage of seeds with a ruptured endosperm compared to the total number of seeds. Additionally, the rate of testa rupture is indicated by the percentage of seeds with a ruptured testa compared to the total number of seeds (optional). Analyze the germination and testa rupture rates by using the IBM SPSS Statistics 22 software. Significant differences in comparison with control seeds (*P < 0.05, **P < 0.01), are analyzed by one-way analysis of variance (ANOVA). The line diagram of seed germination is made by using Origin8.0 software or Microsoft Excel.
Notes
Recipes
Acknowledgments
This protocol was adapted from the Xiaojing Wang Lab at the South China Normal University. This work was supported by the National Natural Science Foundation of China (grant No. 90917011 to X.W.). We thank Dr. craig schluttenhofer (University of Kentucky) for proofreading the article.
References
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