Analysis of T-DNA Integration and Generative Segregation in Transgenic Winter Triticale (x Triticosecale Wittmack)

Overview

Journal BMC Plant Biol

Publisher Biomed Central

Specialty Biology

Date 2012 Sep 26

PMID 23006412

Citations 8

Authors

Goetz Hensel

Sylwia Oleszczuk

Diaa Eldin S Daghma

Janusz Zimny

Michael Melzer

Jochen Kumlehn

Affiliations

Soon will be listed here.

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.

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