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Agrobacterium-mediated transformation of the medicinal plant Centaurea montana

  • Wissam A. Abou-Alaiwi
  • Shobha D. Potlakayala
  • Stephen L. Goldman
  • Puthiyaparambil C. Josekutty
  • Deepkamal N. Karelia
  • Sairam V. Rudrabhatla
Original Paper

Abstract

An efficient transformation system was developed for Centaurea montana by co-cultivation of leaf explants with Agrobacterium tumefaciens strain AGL1 that contained a plasmid harboring the isopentenyl transferase gene under the control of the developmentally regulated Atmyb32 promoter of Arabidopsis thaliana and the gene encoding for hygromycin resistance under the control of the Cauliflower Mosaic Virus 35S (CaMV35S) promoter. A total of 990 explants were infected with Agrobacterium, and 18 shoots were regenerated resulting in an overall transformation efficiency of 1.8%. Molecular analyses, including PCR, Southern blotting and RT-PCR, were performed on T0 and T1 plants to confirm chromosomal integration and expression of the transgene in the phenotypically normal transformed plants. Transformation of C. montana was also performed using A. tumefaciens supervirulent strain EHA105 harboring the β-glucuronidase (GUS) reporter gene. Expression of the GUS gene in the putative transgenics was confirmed using a histochemical GUS assay.

Keywords

Centaurea montana Agrobacterium-mediated transformation Isopentenyl transferase Atmyb32 Delayed senescence 

Notes

Acknowledgments

We would like to acknowledge the funding for this work through the USDA-American University of Beirut grant. We are thankful to Professor German Spangenberg who kindly provided us with the Atmyb32-ipt construct and for his guidance. The authors would like to thank Ms Julianne Dauber and Danielle LaFleur-Do for proof reading the manuscript.

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Copyright information

© Springer Science+Business Media B.V. 2011

Authors and Affiliations

  • Wissam A. Abou-Alaiwi
    • 1
  • Shobha D. Potlakayala
    • 2
  • Stephen L. Goldman
    • 1
  • Puthiyaparambil C. Josekutty
    • 2
  • Deepkamal N. Karelia
    • 2
  • Sairam V. Rudrabhatla
    • 1
    • 2
  1. 1.Plant Science Research CenterUniversity of ToledoToledoUSA
  2. 2.Penn State UniversityMiddletownUSA

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