Abstract
A new selection system for onion transformation that does not require the use of antibiotics or herbicides was developed. The selection system used the Escherichia coli gene that encodes phosphomannose isomerase (pmi). Transgenic plants carrying the manA gene that codes for pmi can detoxify mannose-6-phosphate by conversion to fructose-6-phosphate, an intermediate of glycolysis, via the pmi activity. Six-week-old embryogenic callus initiated from seedling radicle was used for transformation. Transgenic plants were produced efficiently with transformation rates of 27 and 23% using Agrobacterium and biolistic system, respectively. Untransformed shoots were eliminated by a stepwise increase from 10 g l−1 sucrose with 10 g l−1 mannose in the first selection to only10 g l−1 mannose in the second selection. Integrative transformation was confirmed by PCR, RT-PCR and Southern hybridization.
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Abbreviations
- pmi :
-
Phosphomannose isomerase
- 2,4-D :
-
2,4-Dichlorophenoxy acetic acid
- ABA :
-
Abscisic acid
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Acknowledgements
We thank the Korean Federation of Science and Technology, Republic of Korea, for the financial support under ‘Brain pool project’ and ICAR, Government of India, for the study leave for C. Aswath. We also thank Syngenta seeds, Switzerland for sparing the pmi construct. We would like to acknowledge Dr Tim Conner for critically reviewing the manuscript.
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Communicated by I. S. Chung
An erratum to this article can be found at http://dx.doi.org/10.1007/s00299-005-0105-2
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Aswath, C.R., Mo, S.Y., Kim, D.H. et al. Agrobacterium and biolistic transformation of onion using non-antibiotic selection marker phosphomannose isomerase. Plant Cell Rep 25, 92–99 (2006). https://doi.org/10.1007/s00299-005-0022-4
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DOI: https://doi.org/10.1007/s00299-005-0022-4