title: Analysis of T-DNA integration and generative segregation in transgenic winter triticale  (x Triticosecale Wittmack).
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.
authors.affiliation: Leibniz Institute of Plant Genetics and Crop Plant Research (IPK), Plant Reproductive Biology, Corrensstr, 3, 06466, Gatersleben, Germany. hensel@ipk-gatersleben.de
authors.forename: Goetz
authors.initials: G
authors.lastname: Hensel
authors.forename: Sylwia
authors.initials: S
authors.lastname: Oleszczuk
authors.forename: Diaa Eldin S
authors.initials: DE
authors.lastname: Daghma
authors.forename: Janusz
authors.initials: J
authors.lastname: Zimny
authors.forename: Michael
authors.initials: M
authors.lastname: Melzer
authors.forename: Jochen
authors.initials: J
authors.lastname: Kumlehn
mesh_terms: D016960:Agrobacterium tumefaciens / Q000187:drug effects / Q000502:physiology; D020090:Chromosome Segregation / Q000235:genetics*; D002934:Cinnamates / Q000494:pharmacology; D003433:Crosses, Genetic; D004269:DNA, Bacterial / Q000235:genetics*; D002523:Edible Grain / Q000187:drug effects / Q000196:embryology / Q000235:genetics* / Q000382:microbiology; D015870:Gene Expression / Q000187:drug effects; D017930:Genes, Reporter; D005822:Genetic Vectors / Q000235:genetics; D018525:Germination / Q000187:drug effects / Q000235:genetics; D049452:Green Fluorescent Proteins / Q000378:metabolism; D006921:Hygromycin B / Q000031:analogs & derivatives / Q000494:pharmacology; D016254:Mutagenesis, Insertional / Q000235:genetics*; D030821:Plants, Genetically Modified; D012621:Seasons; D012639:Seeds / Q000187:drug effects / Q000235:genetics / Q000254:growth & development; D019076:Transgenes
chemical_list: D002934:Cinnamates; D004269:DNA, Bacterial; C036483:T-DNA; D049452:Green Fluorescent Proteins; D006921:Hygromycin B; C026273:hygromycin A