Korean Journal of Breeding Science (한국육종학회지)

The Korean Society of Breeding Science (한국육종학회)
권호 표지

Efficient Phosphinothricin Mediated Selection of Callus Derived from Brachypodium Mature Seed

  • Jeon, Woong Bae (College of Life sciences and Biotechnology, Korea University) ;
  • Lee, Man Bo (College of Life sciences and Biotechnology, Korea University) ;
  • Kim, Dae Yeon (College of Life sciences and Biotechnology, Korea University) ;
  • Hong, Min Jeong (College of Life sciences and Biotechnology, Korea University) ;
  • Lee, Yong Jin (College of Life sciences and Biotechnology, Korea University) ;
  • Seo, Yong Weon (College of Life sciences and Biotechnology, Korea University)
  • Received : 2010.09.09
  • Published : 2010.09.30
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Abstract

Brachypodium distachyon is rapidly emerged in biological study and has been currently used as a model system for genetics and functional studies for crop improvement and biofuel production. Phosphinothricin (PPT) has been widely used as a selectable agent, which raises ammonium content and induces toxicity in non-transformed plant cells. However PPT selection is not much effective on Brachypodium callus consequently reducing transformation efficiency. In order to identify the efficient conditions of PPT selection, calli obtained from mature seeds of Brachypodium (PI 254867) were cultured on the callus inducing medium (CIM) or regeneration medium (ReM) containing serial dilutions of the PPT (0, 2, 5, 10, and 15 mg/l) in dark or light condition. Callus growth and ammonium content of each treatment were measured 2 weeks after the treatment. Although callus growth and ammonium content did not show much difference in CIM, slow callus growth and increased ammonium accumulation were found in ReM. No significant difference of ammonium accumulation in response to PPT was found between dark and light conditions. In order to identify major factors affecting increased ammonium accumulation, callus was cultured on the media in combined with phytohormones (2,4-D or kinetin) and carbon sources (sucrose or maltose) containing with PPT (5 mg/l). The highest ammonium content in callus was found in the kinetin and maltose media.

Keywords

Acknowledgement

Supported by : National Research Foundation of Korea (NRF)

References

  1. Alves SC, Worland B, Thole V, Snape JW, Bevan MW, Vain P. 2009. A protocol for Agrobacterium-mediated transformation of Brachypodium distachyon community standard line Bd21. Nat. Protoc. 4:638-649. https://doi.org/10.1038/nprot.2009.30
  2. Bablak P, Draper J, Davey MR, Lynch PT. 1995. Plant regeneration and micropropagation of Brachypodium distachyon. Plant Cell Tiss. Org. 42:97-107. https://doi.org/10.1007/BF00037687
  3. Christiansen P, Andersen CH, Didion T, Folling M, Nielsen KK. 2005. A rapid and efficient transformation protocol for the grass Brachypodium distachyon. Plant Cell Rep. 23:751-758. https://doi.org/10.1007/s00299-004-0889-5
  4. De Block M, Botterman J, Vandewiele M, Dockx J, Thoen C, Gossele V, Rao Movva N, Thompson C, Van Montagu M, Leemans J. 1987. Engineering herbicide resistance in plants by expression of a detoxifying enzyme. EMBO J. 6(9):2513-2518.
  5. De Block M, De Sonville A, Debrouwer D. 1995. The selection mechanism of phosphinothricin is influenced by the metabolic status of the tissue. Planta. 197:619-626. https://doi.org/10.1007/BF00191569
  6. Draper J, Mur LAJ, Jenkins G, Ghosh-Biswas GC, Bablak P, Hasterok R, Routledge APM. 2001. Brachypodium distachyon. A new model system for functional genomics in grasses. Plant Physiol. 127:1539-1555. https://doi.org/10.1104/pp.010196
  7. Garvin DF, Gu YQ, Hasterok R, Hazen SP, Jenkins G, Mockler TC, Mur LAJ, Vogel JP. 2008. Development of genetic and genomic research resources for Brachypodium distachyon, a new model system for grass crop research. Crop Sci. 48(S1):S69-S84. https://doi.org/10.2135/cropsci2007.04.0208
  8. Krieg LC, Walker MA, Senaratna T, McKersie BD. 1990. Growth, ammonia accumulation and glutamine synthetase activity in alfalfa (Medicago sativa L.) shoots and cell cultures treated with phosphinothricin. Plant Cell Rep. 9:80-83.
  9. Lin CC, Kao CH. 1996. Disturbed ammonium assimilation is associated with growth inhibition of roots in rice seedlings caused by NaCl. Plant Growth Regul. 18:233-238. https://doi.org/10.1007/BF00024387
  10. Linsmaier EM, Skoog F. 1965. Organic growth factor requirements of tobacco tissue cultures. Physiol. Plant. 18:100-127. https://doi.org/10.1111/j.1399-3054.1965.tb06874.x
  11. Opanowicz M, Vain P, Draper J, Parker D, Doonan JH. 2008. Brachypodium distachyon: making hay with a wild grass. Trends Plant Sci. 13(4):172-177. https://doi.org/10.1016/j.tplants.2008.01.007
  12. Pacurar DI, Thordal-Christensen H, Nielsen KK, Lenk I. 2008. A high-throughput Agrobacterium-mediated transformation system for the grass model species Brachypodium distachyon L. Transgenic Res. 17:965-975. https://doi.org/10.1007/s11248-007-9159-y
  13. Rathore KS, Chowdhury VK, Hodges TK. 1993. Use of bar as a selectable marker gene and for the production of herbicide-resistant rice plants from protoplasts. Plant Mol. Biol. 21:871-884. https://doi.org/10.1007/BF00027118
  14. Tachibana K, Watanabe T, Sekizawa Y, Takematsu T. 1986. Accumulation of ammonia in plants treated with bialaphos. J. Pesticide Sci. 11:33-37. https://doi.org/10.1584/jpestics.11.33
  15. Tsai YC, Kao CH. 2001. Light-dependent ammonium ion toxicity of rice leaves in response to phosphinothricin treatment. Biol. Plant. 45:569-573.
  16. Vanjildorj E, Bae TW, Song IJ, Kim KM, Lim YP, Lee HY. 2008. Herbicide-resistant Transgenic Mongolian Bentgrass (Agrostis mongolica Roshev.) obtained by Agrobacterium-mediated transformatiom. Korean J. Breed. Sci. 40(2):128-135.
  17. Vega-Sanchez ME, Ronald PC. 2010. Genetic and biotechnological approaches for biofuel crop improvement. Curr. Opin. Biotech. 21(2):218-224. https://doi.org/10.1016/j.copbio.2010.02.002
  18. Vogel JP, Garvin DF, Leong OM, Hayden DM. 2006. Agrobacterium-mediated transformation and inbred line development in the model grass Brachypodium distachyon. Plant Cell Tiss. Org. 84:199-211. https://doi.org/10.1007/s11240-005-9023-9