Int. J. Dev. Biol. 57: 483 - 494 (2013)

https://doi.org/10.1387/ijdb.130232mv

Vol 57, Issue 6-7-8

Special Issue: Plant Transgenesis

Higher plant transformation: principles and molecular tools

Open Access | Review | Published: 25 September 2013

Sylvester Anami¹, Elizabeth Njuguna^2,3, Griet Coussens^2,3, Stijn Aesaert^2,3 and Mieke Van Lijsebettens*^,2,3

¹Laboratory of Plant Genetics and Systems Biology, Department of Pure and Applied Sciences, Technical University of Mombasa, Mombasa, Kenya, ²Department of Plant Systems Biology, VIB, Gent, Belgium and ³Department of Plant Biotechnology and Bioinformatics, Ghent University, Gent, Belgium

Abstract

In higher plants, genetic transformation, which is part of the toolbox for the study of living organisms, had been reported only 30 years ago, boosting basic plant biology research, generating superior crops, and leading to the new discipline of plant biotechnology. Here, we review its principles and the corresponding molecular tools. In vitro regeneration, through somatic embryogenesis or organogenesis, is discussed because they are prerequisites for the subsequent Agrobacterium tumefaciens-mediated transferred (T)-DNA or direct DNA transfer methods to produce transgenic plants. Important molecular components of the T-DNA are examined, such as selectable marker genes that allow the selection of transformed cells in tissue cultures and are used to follow the gene of interest in the next generations, and reporter genes that have been developed to visualize promoter activities, protein localizations, and protein-protein interactions. Genes of interest are assembled with promoters and termination signals in Escherichia coli by means of GATEWAY-derived binary vectors that represent the current versatile cloning tools. Finally, future promising developments in transgene technology are considered.

Keywords

Agrobacterium tumefaciens, T-DNA, transgene, plant transformation, somatic embryogenesis, organogenesis