Abstract
Brachypodium distachyon is emerging as the model species for temperate monocotyledonous grasses of the Pooideae, and the genome of the B. distachyon community inbred line Bd21 has recently been sequenced. Here, we report the development of a procedure for the efficient establishment of a cell suspension culture derived from calli. We show that embryogenic potential is maintained in 3-month-old cultures as the cells were positively labelled by the monoclonal antibody, JIM8 which recognizes a carbohydrate epitope often present in arabinogalactan proteins found in the cell walls of cells in embryogenic cultures. Additionally, we were able to regenerate plants from these cell suspension cultures. The cell suspension culture we have established can also be used in studies on plant programmed cell death (PCD). Our results clearly demonstrate that B. distachyon cells can undergo apoptosis-like PCD (AL-PCD) as visualised by the characteristic retraction of the protoplast from the cell wall. As B. distachyon is genetically related to important temperate cereal grass crops like wheat and barley, the ability to utilise cell suspension cultures of B. distachyon to dissect the underlying mechanisms PCD will have important implications for understanding developmental processes in economically important cereal crops.
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Alves, S.C., Worland, B., Thole, V., Snape, J.W., Bevan, M.W., Vain, P. 2009. A protocol for Agrobacterium-mediated transformation of Brachypodium distachyon community standard line Bd21. Nat. Protoc. 4:638–649.
Bossolini, E., Wicker, T., Knobel, P.A., Keller, B. 2007. Comparison of orthologous loci from small grass genomes Brachypodium and rice: Implications for wheat genomics and grass genome annotation. Plant J. 49:704–717.
Draper, J., Mur, L.A.J., Jenkins, G., Ghosh-Biswas, G.C., Bablak, P., Hasterok, R., Routledge, A.P.M. 2001. Brachypodium distachyon. A new model system for function genomics in grasses. Plant Physiol. 127:1539–1555.
Doyle, S.M., Diamond, M., McCabe, P.F. 2010. Chloroplast and reactive oxygen species involvement in apoptotic-like programmed cell death in Arabidopsis suspension cultures. J. Exp. Bot. 61:473–482.
García-Heredia, J.M., Hervás, M., De la Rosa, M.A., Navarro, J.A. 2008. Acetysalicylic acid induces programmed cell death in Arabidopsis cell cultures. Planta 228:89–97.
Garvin, D.F. 2007a. Brachypodium: A new monocot model plant system emerges. J. Sci. Food Agri. 87:1177–1179.
Garvin, D.F. 2007b. Brachypodium distachyon: A new model system for structural and functional analysis of grass genomes. In: Varshney, R.K., Koebner, R.M. (eds), Model Plants and Crop Improvement. Taylor & Francis Press, Florida, USA, pp. 109–123.
Hellwig, S., Drossard, J., Twyman, R.M., Fischer, R. 2004. Plant cell cultures for the production of recombinant proteins. Nat. Biotechnol. 22:1415–1422.
Hong, S.Y., Kwon, T.H., Lee, J.H., Jang, Y.S., Yang, M.S. 2002. Production of biologically active hG-CSF by transgenic plant cell suspension culture. Enzyme Microb. Tech. 6:763–767.
Jeong, C.S., Murthy, H.N., Hahn, E.J., Paek, K.Y. 2008. Improved production of ginsenosides in suspension cultures of ginseng by medium replenishment strategy. J. Biosci. Bioeng. 105:288–291.
Kim, T.G., Baek, M.Y., Lee, E.K., Kwon, T.H., Yang, M.S. 2008. Expression of human growth hormone in transgenic rice cell suspension culture. Plant Cell Rep. 27:885–891.
Lam, E. 2004. Controlled cell death, plant survival and development. Nat. Rev. Mol. Cell Bio. 5:305–315.
Liu, C.Z., Saxena, P.K. 2009. Saussurea medusa cell suspension cultures for flavonoid production. Methods Mol. Biol. 547:53–59.
McCabe, P.F., Leaver, C.J. 2000. Programmed cell death in cell cultures. Plant Mol. Biol. 44:359–368.
McCabe, P.F., Valentine, T.A., Forsberg, L.S., Pennell, R.I. 1997. Soluble signals from cells identified at the cell wall establish a developmental pathway in carrot. Plant Cell 9:2225–2241.
Opanowicz, M., Vain, P., Draper, J., Parker, D., Doonan, J.H. 2008. Brachypodium distachyon: making hay with wild grass. Trends Plant Sci. 13:172–177.
Păcurar, D.I., Thordal-Christensen, H., Nielsen, K.K., Lenk, I. 2007. A high-throughput Agrobacterium-mediated transformation system for the grass model species Brachypodium distachyon L. Transgenic Res. 17:965–975.
Pauk, J., Kertész, Z., Jenes, B., Purnhauser, L., Manninen, O., Pulli, S., Barabás, Z., Dudits, D. 1994. Fertile wheat (Triticum aestivum L.) regenerants from protoplasts of embryogenic suspension culture. Plant Cell Tiss. Org. 38:1–10.
Reape, T.J., McCabe, P.F. 2008. Apoptotic-like programmed cell death in plants. New Phytol. 180:13–26.
Reape, T.J., Molony, E.M., McCabe, P.F. 2008. Programmed cell death in plants: Distinguishing between different modes. J. Exp. Bot. 59:435–444.
Seifert, G.J., Roberts, K. 2007. The biology of arabinogalactan proteins. Ann. Rev. Plant Biol. 58:137–161.
The International Brachypodium Initiative. 2010. Genome sequencing and analysis of the model grass Brachypodium distachyon. Nature 463:763–768.
Thole, V., Alves, S.C., Worland, B., Bevan, M.W., Vain, P. 2009. A protocol for efficiently retrieving and characterizing flanking sequence tags (FSTs) in Brachypodium distachyon T-DNA insertional mutants. Nat. Protoc. 4:650–661.
Vain, P., Worland, B., Thile, V., McKenzie, N., Alves, S.C., Opanowicz, M., Fish, L.J., Bevan, M.W., Snape, J.W. 2008. Agrobacterium-mediated transformation of the temperate grass Brachypodium distachyon (genotype Bd21) for T-DNA insertional mutagenesis. Plant Biotechnol. J. 6:236–245.
Valluri, J.V. 2009. Bioreactor production of secondary metabolites from cell cultures of periwinkle and sandalwood. Methods Mol. Biol. 547:325–335.
van Doorn, W.G., Woltering, E.J. 2005. Many ways to exit? Cell death categories in plants. Trends Plant Sci. 10:117–122.
Vogel, J.P., Garvin, D.F., Leong, O.M., Hayden, D.M. 2006a. Agrobacterium-mediated transformation and inbred line development in the model grass Brachypodium distachyon. Plant Cell Tiss. Org. 84:199–211.
Vogel, J.P., Gu, Y.Q., Twigg, P., Lazo, G.R., Laudencia-Chingcuanco, D., Hayden, D.M., Donze, T.J., Vivian, L.A., Stamova, B., Coleman-Derr, D. 2006b. EST sequencing and phylogenetic analysis of the model grass Brachypodium distachyon. Theor. Appl. Genet. 113:186–195.
Vogel, J.P., Hill, T. 2007. High-efficiency Agrobacterium-mediated transformation of Brachypodium distachyon inbred line Bd21-3. Plant Cell Rep. 27:471–478.
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Hunt, D., Chambers, J.P., Behpouri, A. et al. Brachypodium distachyon Cell Suspension Cultures: Establishment and Utilisation. CEREAL RESEARCH COMMUNICATIONS 42, 58–69 (2014). https://doi.org/10.1556/CRC.2013.0040
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DOI: https://doi.org/10.1556/CRC.2013.0040