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Open Access Research article

Analysis of T-DNA integration and generative segregation in transgenic winter triticale (x Triticosecale Wittmack)

Goetz Hensel1*, Sylwia Oleszczuk2, Diaa Eldin S Daghma34, Janusz Zimny2, Michael Melzer3 and Jochen Kumlehn1

Author affiliations

1 Leibniz Institute of Plant Genetics and Crop Plant Research (IPK), Plant Reproductive Biology, Corrensstr. 3, 06466, Gatersleben, Germany

2 Plant Breeding and Acclimatization Institute, National Research Institute, Radzików, 05-870, Błonie, Poland

3 Leibniz Institute of Plant Genetics and Crop Plant Research (IPK), Structural Cell Biology, Corrensstr. 3, 06466, Gatersleben, Germany

4 National Gene Bank and Genetic Resources, Agriculture Research Center, 12619, Giza, Egypt

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Citation and License

BMC Plant Biology 2012, 12:171  doi:10.1186/1471-2229-12-171

Published: 25 September 2012

Abstract

Background

While the genetic transformation of the major cereal crops has become relatively routine, to date only a few reports were published on transgenic triticale, and robust data on T-DNA integration and segregation have not been available in this species.

Results

Here, we present a comprehensive analysis of stable transgenic winter triticale cv. Bogo carrying the selectable marker gene HYGROMYCIN PHOSPHOTRANSFERASE (HPT) and a synthetic green fluorescent protein gene (gfp). Progeny of four independent transgenic plants were comprehensively investigated with regard to the number of integrated T-DNA copies, the number of plant genomic integration loci, the integrity and functionality of individual T-DNA copies, as well as the segregation of transgenes in T1 and T2 generations, which also enabled us to identify homozygous transgenic lines. The truncation of some integrated T-DNAs at their left end along with the occurrence of independent segregation of multiple T-DNAs unintendedly resulted in a single-copy segregant that is selectable marker-free and homozygous for the gfp gene. The heritable expression of gfp driven by the maize UBI-1 promoter was demonstrated by confocal laser scanning microscopy.

Conclusions

The used transformation method is a valuable tool for the genetic engineering of triticale. Here we show that comprehensive molecular analyses are required for the correct interpretation of phenotypic data collected from the transgenic plants.

Keywords:
Agrobacterium; Winter triticale (x Triticosecale Wittmack); Sexual transmission; Transgene expression; gfp; Copy number