Arabidopsis Hydroponics and Shoot Branching Assay
水培条件下研究拟南芥地上部分枝发育的试验方法
发布: 2012年10月05日第2卷第19期 DOI: 10.21769/BioProtoc.264 浏览次数: 26496
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参见作者原研究论文
The authors used this protocol in:
Apr 2012
Abstract
This protocol establishes growth of Arabidopsis seedlings in a hydroponic medium that is suitable for the controlled addition or withdrawal of any number of nutrients, hormones or metabolites to the plants via the roots. In this case, the described protocol is used to assay sensitivity of shoot branching to exogenously supplied strigolactone, typically the artificial strigolactone GR24. Plants deficient in strigolactone synthesis or response will typically exhibit an increased number of axillary branches compared to wild type. Strigolactone synthesis mutants should respond to exogenous GR24 with reduced numbers and length of axillary shoots, while the phenotype of response mutants will not be rescued.
We present here a more detailed protocol extended from that described in Waters et al. (2012).
Materials and Reagents
- Rock wool/mineral wool (see http://en.wikipedia.org/wiki/Rock_wool; available from hydroponics suppliers)
- GR24, 10 mM in 100% acetone, stored in -20 °C freezer (limited availability; can be purchased from Chiralix: http://www.chiralix.com/rightclick.cfm?id=61574)
- Acetone
- MilliQ water
- Ca(NO3)2
- KNO3
- NH4NO3
- MgSO4.7H2O
- KH2PO4
- KCl
- H3BO3
- MnCl2.4H2O
- ZnSO4.7 H2O
- CuSO4.5 H2O
- CoCl2.6 H2O
- (NH4)6Mo7O24.4 H2O
- Fe-EDTA
- 2-(N-morpholino) ethanesulfonic acid, a buffering agent (MES)
- 0.5x Hoagland’s nutrient solution (pH 5.9) (see Recipes)
- Hoagland’s solutions (see Recipes)
Equipment
- Controlled environment growth chamber or room
- Sealed plastic box for growing plants, with transparent lid and with rack to hold 1.5 ml microcentrifuge tubes, e.g. I5100-43, Astral Scientific, NSW, Australia. Also available from:
http://www.bioexpress.com/divinity-cart/item/502100/GeneMate-Floater-Microtube-Rack/1.html
http://www.labsource.com/Catalog/Group.aspx?GroupID=1455 - Black 1.5 ml microcentrifuge tubes (opaque tubes minimizes light transmission and hence algal growth)
- Fine forceps
- Cork borer, 8 mm diameter, to produce cylinders of rock wool (see http://en.wikipedia.org/wiki/Cork_borer)
Figure 1. Three-week-old Arabidopsis Ler plants grown in hydroponics according to this protocol. The lid of the box has been removed in this photograph. Grid squares are 1 cm.
Procedure
文章信息
版权信息
© 2012 The Authors; exclusive licensee Bio-protocol LLC.
如何引用
Waters, M. T., Bussell, J. D. and Jost, R. (2012). Arabidopsis Hydroponics and Shoot Branching Assay. Bio-protocol 2(19): e264. DOI: 10.21769/BioProtoc.264.
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分类
植物科学 > 植物生理学 > 植物生长
植物科学 > 植物发育生物学 > 形态建成
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