Abstract
Transgenic torenia plants were obtained using the selectable marker gene phosphomannose isomerase (manA), which encodes the enzyme phosphomannose isomerase (PMI) to enable selection of transformed cells on media containing mannose. We found that shoot organogenesis in torenia leaf explants was effectively suppressed on medium supplemented with mannose, which indicated that torenia cells had little or no PMI activity and could not utilize mannose as a carbon source. Leaf pieces from in vitro-germinated plants were inoculated with Agrobacterium tumefaciens EHA105 containing the binary vector pKPJ with both hpt and ManA genes, and subsequently selected on shoot induction (SI) medium (half strength MS basal + 4.4 μM BA + 0.5 μM NAA) supplemented with 20 g l−1 mannose and 5 g l−1 sucrose as carbon sources. Transformed plants were confirmed by PCR and Southern blot. The transgene expression was evaluated using Northern blot and the chlorophenol red assay. The transformation efficiency ranged from 7% to 10%, which is 1–3% higher than that obtained by selection with hygromycin. This system provides an efficient manner for selecting transgenic flower plants without using antibiotics or herbicides.
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Abbreviations
- 2,4-D:
-
2,4-Dichlorophenoxyacetic acid
- BA:
-
Benzyladenine
- IBA:
-
Indole-3-butyric acid
- MS:
-
Murashige and Skoog’s salts
- NAA:
-
α-Naphthaleneacetic acid
- PMI:
-
Phosphomannose isomerase
- AS:
-
Acetosyringone
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Acknowledgement
This research was supported by the National Natural Science Foundation of China (No: 30370137) and a project from Guandong Province (2006A20101007). We thank Dr. R.A. Jefferson, CAMBIA, Canberra, Australia, for kindly providing pCAMBIA1390, pCAMBIA1301. We would also like to thank the anonymous referees for their useful comments and valuable suggestions to improve the paper.
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Li, HQ., Kang, PJ., Li, ML. et al. Genetic transformation of Torenia fournieri using the PMI/mannose selection system. Plant Cell Tiss Organ Cult 90, 103–109 (2007). https://doi.org/10.1007/s11240-007-9260-1
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DOI: https://doi.org/10.1007/s11240-007-9260-1