Efficient Phosphinothricin Mediated Selection of Callus Derived from Brachypodium Mature Seed
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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.
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.
참고문헌 (18)
- Alves SC, "A protocol for Agrobacterium-mediated transformation of Brachypodium distachyon community standard line Bd21", Nat. Protoc., 4, pp.638~649 (2009)
- Bablak P, "Plant regeneration and micropropagation of Brachypodium distachyon", Plant Cell Tiss. Org., 42, pp.97~107 (1995)
- Christiansen P, "A rapid and efficient transformation protocol for the grass Brachypodium distachyon", Plant Cell Rep., 23, pp.751~758 (2005)
- De Block M, "Engineering herbicide resistance in plants by expression of a detoxifying enzyme", EMBO J., 6(9), pp.2513~2518 (1987)
- De Block M, "The selection mechanism of phosphinothricin is influenced by the metabolic status of the tissue", Planta., 197, pp.619~626 (1995)
- Draper J, "Brachypodium distachyon. A new model system for functional genomics in grasses", Plant Physiol., 127, pp.1539~1555 (2001)
- Garvin DF, "Development of genetic and genomic research resources for Brachypodium distachyon, a new model system for grass crop research", Crop Sci., 48(S1), pp.S69~S84 (2008)
- Krieg LC, "Growth, ammonia accumulation and glutamine synthetase activity in alfalfa (Medicago sativa L.) shoots and cell cultures treated with phosphinothricin", Plant Cell Rep., 9, pp.80~83 (1990)
- Lin CC, "Disturbed ammonium assimilation is associated with growth inhibition of roots in rice seedlings caused by NaCl", Plant Growth Regul., 18, pp.233~238 (1996)
- Linsmaier EM, "Organic growth factor requirements of tobacco tissue cultures", Physiol. Plant., 18, pp.100~127 (1965)
- Opanowicz M, "Brachypodium distachyon: making hay with a wild grass", Trends Plant Sci., 13(4), pp.172~177 (2008)
- Pacurar DI, "A high-throughput Agrobacterium-mediated transformation system for the grass model species Brachypodium distachyon L", Transgenic Res., 17, pp.965~975 (2008)
- Rathore KS, "Use of bar as a selectable marker gene and for the production of herbicide-resistant rice plants from protoplasts", Plant Mol. Biol., 21, pp.871~884 (1993)
- Tachibana K, "Accumulation of ammonia in plants treated with bialaphos", J. Pesticide Sci., 11, pp.33~37 (1986)
- Tsai YC, "Light-dependent ammonium ion toxicity of rice leaves in response to phosphinothricin treatment", Biol. Plant., 45, pp.569~573 (2001)
- Enkhchimeg Vanjildorj, "Herbicide-resistant Transgenic Mongolian Bentgrass (Agrostis mongolica Roshev.) obtainedby Agrobacterium-mediated Transformation", 한국육종학회지, 40(2), pp.128~135 (2008), 한국육종학회
- Vega-Sanchez ME, "Genetic and biotechnological approaches for biofuel crop improvement", Curr. Opin. Biotech., 21(2), pp.218~224 (2010)
- Vogel JP, "Agrobacterium-mediated transformation and inbred line development in the model grass Brachypodium distachyon", Plant Cell Tiss. Org., 84, pp.199~211 (2006)