Abstract
Brachypodium distachyon is a model grass species for economically important cereal crops. Efforts are in progress to develop useful functional genomic resources in Brachypodium. A tobacco retrotransposon, Tnt1, has been used successfully in recent past to generate insertional mutagenesis in several dicot plant species. Tnt1 retrotransposon replicates, transposes, and inserts at multiple random genomic locations in the plant genome. Transposition occurs only during somatic embryogenesis but not during seed transmission. We developed Brachypodium transgenic plants that can express the Tnt1 element. Here, we describe an efficient tissue culture-based approach to generate Tnt1 insertional mutant population using transgenic Brachypodium line expressing the Tnt1 retrotransposon.
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References
Brkljacic J, Grotewold E, Scholl R et al (2011) Brachypodium as a model for the grasses: today and the future. Plant Physiol 157:3–13
Brutnell TP, Bennetzen JL, Vogel JP (2015) Brachypodium distachyon and Setaria viridis: model genetic systems for the grasses. Ann Rev Plant Biol 66:465–485
The International Brachypodium Initiative (2010) Genome sequencing and analysis of the model grass Brachypodium distachyon. Nature 463:763–768
Bragg JN, Wu J, Gordon SP, Guttman MA, Thilmony RL, Lazo GR, Gu YQ, Vogel JP (2012) Generation and characterization of the Western Regional Research Center Brachypodium T-DNA insertional mutant collection. PLoS One 7:e41916
Vain P, Worland B, Thole V, McKenzie N, Opanowicz M, Fish LJ, Bevan MW, Snape JW (2008) Agrobacterium-mediated transformation of the temperate grass Brachypodium distachyon (genotype Bd21) for T-DNA insertional mutagenesis. Plant Biotechnol J 6:236–245
Courtial B, Feuerbach F, Eberhard S, Rohmer L, Chiapello H, Camilleri C, Lucas H (2001) Tnt1 transposition events are induced by in vitro transformation of Arabidopsis thaliana, and transposed copies integrate into genes. Mol Genet Genomics 265:32–42
Cui Y, Barampuram S, Stacey MG, Hancock CN, Findley S, Mathieu M, Zhang ZY, Parrott WA, Stacey G (2013) Tnt1 retrotransposon mutagenesis: a tool for soybean functional genomics. Plant Physiol 161:36–47
d’Erfurth I, Cosson V, Eschstruth A, Lucas H, Kondorosi A, Ratet P (2003) Efficient transposition of the Tnt1 tobacco retrotransposon in the model legume Medicago truncatula. Plant J 34:95–106
Duangpan S, Zhang W, Wu Y, Jansky SH, Jiang J (2013) Insertional mutagenesis using Tnt1 retrotransposon in potato. Plant Physiol 163:21–29
Mazier M, Botton E, Flamain F, Bouchet JP, Courtial B, Chupeau MC, Chupeau Y, Maisonneuve B, Lucas H (2007) Successful gene tagging in lettuce using the Tnt1 retrotransposon from tobacco. Plant Physiol 144:18–31
Tadege M, Wen JQ, He J, Tu HD, Kwak Y, Eschstruth A, Cayrel A, Endre G, Zhao PX, Chabaud M et al (2008) Large-scale insertional mutagenesis using the Tnt1 retrotransposon in the model legume Medicago truncatula. Plant J 54:335–347
Grandbastien MA, Lucus H, Morel JB, Mhiri C, Vernhettes S, Casacubert JM (1997) The expression of the tobacco Tnt1 retrotransposon is linked to the plant defense responses. Genetica 100:241–252
Bragg JN, Anderton A, Nieu R, Vogel JP (2015) Brachypodium distachyon. In: Wang K (ed) Agrobacterium protocols, Methods in molecular biology, vol 1223. Springer, New York, pp 17–33
Vogel JP, Hill T (2008) High-efficiency Agrobacterium-mediated transformation of Brachypodium distachyon inbred line Bd21-3. Plant Cell Rep 27:471–478
Acknowledgement
We thank Janie Gallaway and Colleen Elles for taking care of the plants in the greenhouse. This work was supported by the Noble Research Institute, LLC.
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Gill, U.S., Serrani-Yarce, J.C., Lee, HK., Mysore, K.S. (2018). Tissue Culture (Somatic Embryogenesis)-Induced Tnt1 Retrotransposon-Based Mutagenesis in Brachypodium distachyon . In: Sablok, G., Budak, H., Ralph, P. (eds) Brachypodium Genomics. Methods in Molecular Biology, vol 1667. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-7278-4_5
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DOI: https://doi.org/10.1007/978-1-4939-7278-4_5
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