biologia plantarum

International journal on Plant Life established by Bohumil Němec in 1959

Biologia plantarum 54:777-780, 2010 | DOI: 10.1007/s10535-010-0141-9

Efficient biolistic transformation of the moss Physcomitrella patens

M. ©mídková1, M. Holá1, K. J. Angelis1,*
1 Institute of Experimental Botany, Academy of Sciences of the Czech Republic, Prague, Czech Republic

High rates of homologous recombination (HR) in comparison to other plants make the moss Physcomitrella patens an attractive model organism for genetic studies as well as biotechnological applications. We describe a simple protocol for the efficient biolistic transformation of protonemal tissue with minimum tissue handling steps. The transformation efficiency depends on the biolistic conditions. The bombardment of tissue with 1 μm gold particles yielded between 20 and 40 stable transformants per 1 μg of DNA. Transformation with circular plasmids generates higher frequencies of random transgene integration, whereas linear plasmids are more efficient in generating gene-targeted insertions.

Keywords: Helios biolistic gun; moss protonemal tissue; particle size; random and targeted integration; regeneration
Subjects: particle bombardment; Physcomitrella patens; Southern blot analysis; transgenic plants

Received: July 1, 2009; Accepted: September 7, 2009; Published: December 1, 2010  Show citation

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©mídková, M., Holá, M., & Angelis, K.J. (2010). Efficient biolistic transformation of the moss Physcomitrella patens. Biologia plantarum, 54(4), 777-780. doi: 10.1007/s10535-010-0141-9
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    References

    1. Bezanilla, M., Pan, A., Quatrano, R.S.: RNA interference in the moss Physcomitrella patens. - Plant Physiol. 133; 470-474, 2003. Go to original source...
    2. Cho, S.H., Chung, Y.S., Cho, S.K., Rim, Y.W., Shin, J.S.: Particle bombardment mediated transformation and GFP expression in the moss Physcomitrella patens. - Mol. Cells 9; 14-19, 1999.
    3. Cove, D.J.: The moss Physcomitrella patens. - Annu. Rev. Genet. 39; 339-358, 2005. Go to original source...
    4. Frank, W., Decker, E.L., Reski, R.: Molecular tools to study Physcomitrella patens. - Plant Biol. 7; 220-227, 2005. Go to original source...
    5. Gal-On, A., Meiri, E., Elman, C., Gray, E., Gaba, V.: Simple hand-held devices for the efficient infection of plants with viral-encoding constructs by particle bombardment. - J. Virol. Meth. 64; 103-110, 1997. Go to original source...
    6. Gray, D.J., Hiebert, E., Lin, C.M., Compton, M.E., McColley, D.W., Harrison, R.J., Gaba, V.P.: Simplified construction and performance of a device for particle bombardment. - Plant Cell Tissue Organ Cult. 37; 179-184, 1994. Go to original source...
    7. Knight, C.D., Cove, D.J., Cuming, A.C., Quatrano, R.S.: Moss gene technology. - In: Gilmartin, P.M., Bowler, C. (ed.): Molecular Plant Biology. Vol. 2A Practical Approach. Pp. 285-301. Oxford University Press, Oxford 2002.
    8. Sawahel, W., Onde, S., Knight, C.D., Cove, D.J.: Transfer of foreign DNA into Physcomitrella patens protonemal tissue by using the gene gun. - Plant mol. Biol. Rep. 10; 315-316, 1992. Go to original source...
    9. Schaefer, D.G.: A new moss genetics: targeted mutagenesis in Physcomitrella patens. - Annu. Rev. Plant Biol. 53; 477-501, 2002. Go to original source...
    10. Schaefer, D.G., Zrÿd, J.-P., Knight, C.D., Cove, D.J.: Stable transformation of the moss Physcomitrella patens. - Mol. gen. Genet. 226; 418-424, 1991. Go to original source...

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