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RESEARCH ARTICLE
Contents Vol 33(7)

Transformation of cambial tissue in vivo provides an efficient means for induced somatic sector analysis and gene testing in stems of woody plant species

Kim S. Van Beveren A , Antanas V. Spokevicius A , Josquin Tibbits A , Qing Wang A and Gerd Bossinger A B
+ Author Affiliations
- Author Affiliations

A School of Forest and Ecosystem Science, The University of Melbourne, Water Street, Creswick, Vic. 3363, Australia.

B Corresponding author. Email: gerd@unimelb.edu.au

Functional Plant Biology 33(7) 629-638 https://doi.org/10.1071/FP06057
Submitted: 17 March 2006  Accepted: 28 April 2006   Published: 3 July 2006

Abstract

Large-scale functional analysis of genes and transgenes suspected to be involved in wood development in trees is hindered by long generation times, low transformation and regeneration efficiencies and difficulties with phenotypic assessment of traits, especially those that appear late in a tree’s development. To avoid such obstacles many researchers have turned to model plants such as Arabidopsis thaliana (L.) Heynh., Zinnia elegans Jacq. and Nicotiana ssp., or have focused their attention on in vitro wood formation systems or in vivo approaches targeting primary meristems for transformation. Complementing such efforts, we report the use of Agrobacterium to introduce transgenes directly into cambial cells of glasshouse-grown trees in order to create transgenic somatic tissue sectors. These sectors are suitable for phenotypic evaluation and analysis of target gene function. In our experiments the wood formation zone containing the cambium of Eucalyptus, Populus and Pinus species of varying age was inoculated with Agrobacterium containing a CaMV 35S::GUS construct. Following an initial wound response, frequent and stable transformation was observed in the form of distinct GUS-staining patterns (sectors) in newly formed secondary tissues. Sector size and extent depended on the cell type transformed, the species and the length of time treated plants were allowed to grow (more than two years in some cases). Induced somatic sector analysis (ISSA) can now be efficiently used to study cell fate and gene function during secondary growth in stems of forest tree species.

Keywords: Agrobacterium, cambium, Eucalyptus, in vivo transformation, Pinus, Populus, secondary growth, wood formation.


Acknowledgments

We thank Simon Southerton, Zander Myburg and Rick Meilan for critically reading our manuscript and for many helpful comments.


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