Molecular analysis of transgene and vector backbone integration into the barley genome following Agrobacterium-mediated transformation

Plant Cell Reports
Mette LangePreben B Holm

Abstract

We report a large-scale study on the frequency of transgene and T-DNA backbone integration following Agrobacterium-mediated transformation of immature barley embryos. One hundred and ninety-one plant lines were regenerated after hygromycin selection and visual selection for GFP expression at the callus stage. Southern blotting performed on a subset of 53 lines that were PCR positive for the GFP gene documented the integration of the GFP gene in 27 of the lines. Twenty-three of these lines expressed GFP in T(1) plantlets. Southern blotting with a vector backbone probe revealed that 13 of the 27 lines possessed one or more vector backbone fragments illustrating the regular occurrence of vector backbone integration following Agrobacterium infection of barley immature embryos.

References

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Citations

Jun 24, 2010·Plant Cell Reports·Massimo ConfalonieriAlma Balestrazzi
Jan 13, 2011·Molecular Biotechnology·Danny CullenMark Taylor
Jul 16, 2013·Trends in Biotechnology·Audur MagnusdottirBjörn Larus Örvar
Jul 9, 2009·International Journal of Plant Genomics·Goetz HenselJochen Kumlehn
Oct 2, 2007·Journal of Plant Physiology·Goetz HenselJochen Kumlehn
Oct 1, 2011·Plant Biotechnology Journal·Inger B HolmePreben B Holm
Feb 21, 2013·Plant Biotechnology Journal·Inger Bæksted HolmePreben Bach Holm
Apr 27, 2011·Molecular Ecology Resources·Klaus GaseIan T Baldwin
Dec 26, 2012·Gene·Shweta Mehrotra, Vinod Goyal
Oct 8, 2013·Plant Science : an International Journal of Experimental Plant Biology·Joseph G DubouzetArmin Wagner
Sep 13, 2017·International Journal of Molecular Sciences·Alessandro NicoliaDaniele Rosellini
Jun 1, 2016·Journal, Genetic Engineering & Biotechnology·Bipinchandra B Kalbande, Anita S Patil
Dec 17, 2017·Journal of Integrative Plant Biology·Wai Li LimRachel A Burton

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