An Agrobacterium -mediated transient gene expression system for intact leaves

An Agrobacterium -mediated transient gene expression system for intact leaves An efficient and reproducible Agrobacterium -mediated transient gene expression system for intact leaf tissue was developed. A high level of transient expression was observed when bacteria, which were pretreated in vir gene-inducing conditions, were infiltrated into complete leaf tissue. Histochemical β-glucuronidase assays showed large transgene-expressing sectors comprising of up to 90% of the leaf area. As a consequence of infiltration, the induced bacteria entered into the intercellular spaces, thus cnabling T-DNA transfer in all cell layers of the leaf. The protocol was optimized for Phascolus vulgaris leaves, but similar results were obtained with other plant species, such as Phaseolus acutifolius , poplar, and tobacco. A β-glucuronidase chimeric gene interrupted by an intron was used as a marker for histological detection of the sectors. http://www.deepdyve.com/assets/images/DeepDyve-Logo-lg.png Plant Science Elsevier

An Agrobacterium -mediated transient gene expression system for intact leaves

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Publisher
Elsevier
Copyright
Copyright © 1997 Elsevier Science Ireland Ltd. All Rights Reserved.
ISSN
0168-9452
D.O.I.
10.1016/S0168-9452(96)04541-4
Publisher site
See Article on Publisher Site

Abstract

An efficient and reproducible Agrobacterium -mediated transient gene expression system for intact leaf tissue was developed. A high level of transient expression was observed when bacteria, which were pretreated in vir gene-inducing conditions, were infiltrated into complete leaf tissue. Histochemical β-glucuronidase assays showed large transgene-expressing sectors comprising of up to 90% of the leaf area. As a consequence of infiltration, the induced bacteria entered into the intercellular spaces, thus cnabling T-DNA transfer in all cell layers of the leaf. The protocol was optimized for Phascolus vulgaris leaves, but similar results were obtained with other plant species, such as Phaseolus acutifolius , poplar, and tobacco. A β-glucuronidase chimeric gene interrupted by an intron was used as a marker for histological detection of the sectors.

Journal

Plant ScienceElsevier

Published: Jan 15, 1997

References

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