Published: Vol 3, Iss 12, Jun 20, 2013 DOI: 10.21769/BioProtoc.795 Views: 19548
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Abstract
In L. japonicus, hairy root transformation is the very useful technique to generate transformed root systems in a short term. This protocol was previously described (Kumagai and Kouchi, 2003) with some modifications. After the infection of Agrobacterium rhizogenes, L. japonicus develops not only transformed but also untransformed roots. Thus, transgenic roots need to be identified by certain indications. In this protocol, we use the GFP florescent signals as such indication.
Materials and Reagents
Equipment
Procedure
A. Plant growth
B. Culture of Agrobacterium
Streak A. rhizogenes harboring the desired construct on LB plate with appropriate antibodies for 2 days at 28 °C, then spread bacteria all around sterilized dish (9 cm in diameter) and incubate for 1 day.
C. Infection of A. rhizogenes with L. japonicus
D.Induction of hairy roots
Recipes
Acknowledgments
This work was supported by MEXT/JSPS KAKENHI, Japan (22870035, 23012038, 25114519 to Takuya Suzaki).
References
Article Information
Copyright
© 2013 The Authors; exclusive licensee Bio-protocol LLC.
How to cite
Okamoto, S., Yoro, E., Suzaki, T. and Kawaguchi, M. (2013). Hairy Root Transformation in Lotus japonicus. Bio-protocol 3(12): e795. DOI: 10.21769/BioProtoc.795.
Category
Plant Science > Plant transformation > Agrobacterium
Molecular Biology > DNA > Transformation
Cell Biology > Tissue analysis > Tissue staining
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