Arabidopsis Seed Germination Assay with Gibberellic Acid
GA处理条件下的拟南芥种子萌发实验
发布: 2016年11月20日第6卷第22期 DOI: 10.21769/BioProtoc.2005 浏览次数: 18806
评审: Arsalan DaudiYuan ChenAnonymous reviewer(s)
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参见作者原研究论文
The authors used this protocol in:
Nov 2015
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
- 1.5 ml Eppendorf tubes (Eppendorf)
- 950 x 150 mm Petri dish (Shanghai Wuyi Glass Factory, catalog number: 1771 )
- Sterile pipette tips (Corning, Axygen®)
- Parafilm (Bemis, catalog number: PM-996 )
- Aluminum foil (GLAD, catalog number: F5M )
- 0.2 µm syringe filters (Pall, Acrodisc®, catalog number: 4554 )
- Arabidopsis seeds
- Murashige and Skoog basal medium (Sigma-Aldrich, catalog number: M5519 )
- Agar (MBCHEM, catalog number: 170837 , agented by WHIGA in China)
- Sucrose, purity: AR (Tianjin Damao Chemical Reagent Factory, catalog number: 57-50-1 )
- Gibberellic acid (GA3) (Sigma-Aldrich, catalog number: G7645 )
- Ethanol, purity: AR (Tianjin Damao Chemical Reagent Factory, catalog number: 64-17-5 )
- Potassium hydroxide (KOH), purity: AR (Chengdu Institute of Chemical Reagents, catalog number: 1310-73-2 )
- MS solid medium (see Recipes)
- 10 mM GA3 stock (see Recipes)
- 70% ethanol (see Recipes)
- 1% sodium hypochlorite (see Recipes)
- Autoclaved distilled water (see Recipes)
Equipment
- Aquapro water purification system (Aquapro, model: AQ06062001 )
Note: This product has been discontinued. - Refrigerator (Haier, model: BCD-648WDBE )
- Autoclave (HIRAYAMA, model: HVE-50 )
- 1 ml pipette (Eppendorf, catalog number: 3120620.001 )
- Drying oven (ZenithLabo, model: DHG 9070A )
- 100 ml flasks
- Microwave oven (Galanz, model: G70F20N2L-DG )
- Benchtop (Suzhou Antai Air-tech, model: SW-CJ-1FD )
- Stereoscopic microscope (Nikon, model: SMZ1500 )
- Digital camera (10 megapixels) (Nikon, model: COOLPIX4500 )
- Thermometer (WUqiang, China)
Software
- Photoshop CS5 software
- IBM SPSS Statistics 22 software
- Origin8.0 software or Microsoft Excel
Procedure
文章信息
版权信息
© 2016 The Authors; exclusive licensee Bio-protocol LLC.
如何引用
Readers should cite both the Bio-protocol article and the original research article where this protocol was used:
- Zhong, C., Xu, H., Ye, S., Zhang, S. and Wang, X. (2016). Arabidopsis Seed Germination Assay with Gibberellic Acid. Bio-protocol 6(22): e2005. DOI: 10.21769/BioProtoc.2005.
- Zhong, C., Xu, H., Ye, S., Wang, S., Li, L., Zhang, S. and Wang, X. (2015). Gibberellic acid-stimulated Arabidopsis6 serves as an integrator of gibberellin, abscisic acid, and glucose signaling during seed germination in Arabidopsis. Plant Physiol 169(3): 2288-2303.
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分类
植物科学 > 植物生物化学 > 植物激素
植物科学 > 植物生理学 > 植物生长
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