Transgene structures suggest that multiple mechanisms are involved in T-DNA integration in plants

Qian-Hao Zhu; Kerrie Ramm; Andrew Eamens; Elizabeth S Dennis; Narayana M Upadhyaya

doi:10.1016/j.plantsci.2006.03.019

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Transgene structures suggest that multiple mechanisms are involved in T-DNA integration in plants

Journal article

Peer reviewed

Transgene structures suggest that multiple mechanisms are involved in T-DNA integration in plants

Qian-Hao Zhu, Kerrie Ramm, Andrew Eamens, Elizabeth S Dennis and Narayana M Upadhyaya

Plant Science, Vol.171(3), pp.308-322

2006

DOI: https://doi.org/10.1016/j.plantsci.2006.03.019

PMID: 22980200

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http://dx.doi.org/10.1016/j.plantsci.2006.03.019View

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Abstract

Rice

T-DNA integration

T-DNA junction

Plant Biology

Oryza sativa

Molecular Manipulation

Genetic Transformation

Technology Development

To gain further understanding of the mechanisms involved in Agrobacterium-mediated genetic transformation and T-DNA integration, we analysed 156 T-DNA/rice, 69 T-DNA/T-DNA and 11 T-DNA/vector backbone (VB) junctions, which included 171 left borders (LB) and 134 right borders (RB). Conserved cleavage was observed in 6% of the LB and 43% of the RB. Terminal microhomology (1–10bp) was identified in 58% of T-DNA/rice, 43% of T-DNA/T-DNA and 82% of T-DNA/VB junctions, and this occurred particularly at the LB junctions. Approximately 32% of both T-DNA/rice and T-DNA/T-DNA junctions harboured 1–344bp of filler DNA that was derived mainly from the T-DNA region adjacent to the breakpoint and/or from the rice genome flanking the T-DNA integration site. Structure of the filler DNA was more complicated at the T-DNA/T-DNA junction than at the T-DNA/rice junction, indicating the presence of T-DNA recombination or rearrangement prior to or during T-DNA integration. When two T-DNAs were integrated in the inverted repeat configuration, significant truncation was always observed in one of the two T-DNAs whereas with direct repeat configuration, a large truncation was less frequent. Most integration events analysed in this study could be addressed by previously proposed models; however, the characteristics of the T-DNA repeats and the complicated filler DNA between two T-DNA copies imply that multiple mechanisms are involved in the formation of T-DNA repeats as well as in T-DNA integration in plants.

Details

Title: Transgene structures suggest that multiple mechanisms are involved in T-DNA integration in plants
Authors: Qian-Hao Zhu (Author) - CSIRO Plant Industry
Kerrie Ramm (Author) - CSIRO Plant Industry
Andrew Eamens (Author) - CSIRO Plant Industry
Elizabeth S Dennis (Author) - CSIRO Plant Industry
Narayana M Upadhyaya (Author) - CSIRO Plant Industry
Publication details: Plant Science, Vol.171(3), pp.308-322
Publisher: Elsevier Ireland Ltd.
DOI: 10.1016/j.plantsci.2006.03.019
ISSN: 1873-2259
PMID: 22980200
Organisation Unit: School of Health and Behavioural Sciences - Legacy; University of the Sunshine Coast, Queensland; School of Health - Biomedicine
Language: English
Record Identifier: 99656589102621
Output Type: Journal article

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