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The maize Knotted1 gene is an effective positive selectable marker gene for Agrobacterium-mediated tobacco transformation

  • Genetic Transformation and Hybridization
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Abstract

We have assessed the use of a homeobox gene knotted1 (kn1) from maize as a selectable marker gene for plant transformation. The kn1 gene under the control of cauliflower mosaic virus 35S promoter (35S::kn1) was introduced into Nicotiana tabacum cv. Xanthi via Agrobacterium-mediated transformation. Under nonselective conditions (without antibiotic selection) on a hormone-free medium (MS), a large number of transgenic calli and shoots were obtained from explants that were infected with Agrobacterium tumefaciens LBA4404 harboring the 35S::kn1 gene. On the other hand, no calli or shoots were produced from explants that were infected with an Agrobacterium strain harboring pBI121 (nptII selection) or from uninfected controls cultured under identical conditions. Relative to kanamycin selection conferred by nptII, the use of kn1 resulted in a 3-fold increase in transformation efficiency. The transgenic status of shoots obtained was confirmed by both histochemical detection of GUS activity and molecular analysis. The results presented here suggest that kn1 gene could be used as an effective alternative selection marker with a potential to enhance plant transformation efficiency in many plant species. With kn1 gene as a selection marker gene, no antibiotic-resistance or herbicide-resistance genes are needed so that potential risks associated with the use of these traditional selection marker genes can be eliminated.

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Abbreviations

CaMV 35S :

cauliflower mosaic virus 35S

kn1 :

Knotted1 gene

ipt :

Isopentenyl transferase

BA :

Benzylaminopurine

NAA :

Naphthaleneacetic acid

NPTII :

Neomycin phosphotransferase

MS Medium :

Musashige & Skoog medium

GUS :

β-glucuronidase

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Acknowledgements

This research was supported by grants from the National Natural Science Foundation of China (#30300225). The authors would like to thank Dr. Prakash Lakshmanan for his help in the preparation of the manuscript.

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Authors and Affiliations

  1. Biotechnology Research Center, Southwest University, Chongqing, 400716, P.R. China

    Keming Luo, Xuelian Zheng, Yuehua Xiao & Yan Pei

  2. Department of Biotechnology, College of Life Sciences, Hubei University, Wuhan, 430062, P.R. China

    Yongqin Chen

  3. Key Laboratory of Agricultural Biotechnology, Guizhou University, Guizhou, 550025, P.R. China

    Degang Zhao

  4. Department of Plant Science, University of Connecticut, Storrs, CT, 06269, USA

    Keming Luo, Xuelian Zheng, Yongqin Chen, Richard McAvoy & Yi Li

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  1. Keming Luo
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  2. Xuelian Zheng
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  3. Yongqin Chen
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  4. Yuehua Xiao
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  5. Degang Zhao
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  6. Richard McAvoy
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  7. Yan Pei
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  8. Yi Li
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Correspondence to Yan Pei.

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Communicated by P. Lakshmanan

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Luo, K., Zheng, X., Chen, Y. et al. The maize Knotted1 gene is an effective positive selectable marker gene for Agrobacterium-mediated tobacco transformation. Plant Cell Rep 25, 403–409 (2006). https://doi.org/10.1007/s00299-005-0051-z

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  • DOI: https://doi.org/10.1007/s00299-005-0051-z

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