Abstract
Agrobacterium tumefaciens strain EHA105 harboring an ipt-type MAT vector, pNPI132, was used to produce morphologically normal transgenic Nierembergia caerulea cv. Mont Blanc employing ipt gene as the selectable marker gene. β-glucuronidase (GUS) gene was used as model gene of interest. The MAT vector system is a positive selection system that gives the advantage of regeneration to the transgenic cells without killing the non-transgenic cells. Infected explants were cultured on hormone- and antibiotic-free MS medium, and 65% of the regenerated shoots developed ipt shooty phenotype-morphologically abnormal shoots, within approximately 3 months after co-cultivation. Twenty morphologically normal shoots were produced from 12 transgenic ipt shoots 7 months after co-cultivation. The normal shoots rooted well on hormone-free MS medium. Ninety percent of the normal shoots were ipt −, GUS+ and excision+ as determined by PCR and Southern blot analyses. These results indicate that ipt-type MAT vector system can be used successfully in Nierembergia to produce marker-free transgenic plants without using phytohormones and selective chemical agents.
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Abbreviations
- ESP:
-
Extreme shooty phenotype
- GUS:
-
β-Glucuronidase
- ipt :
-
Isopentenyl transferase
- MAT:
-
Multi-auto-transformation
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Acknowledgement
The authors would like to thank Dr. H. Ebinuma, Pulp and Paper Research, Nippon Paper Industries, Tokyo for providing the MAT vector constructs.
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Khan, R.S., Chin, D.P., Nakamura, I. et al. Production of marker-free transgenic Nierembergia caerulea using MAT vector system. Plant Cell Rep 25, 914–919 (2006). https://doi.org/10.1007/s00299-006-0125-6
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DOI: https://doi.org/10.1007/s00299-006-0125-6