Analysis of T-DNA integration and generative segregation in transgenic winter triticale (x Triticosecale Wittmack).
BMC Plant Biol
; 12: 171, 2012 Sep 25.
Article
en En
| MEDLINE
| ID: mdl-23006412
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.
Texto completo:
1
Colección:
01-internacional
Base de datos:
MEDLINE
Asunto principal:
Estaciones del Año
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ADN Bacteriano
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Grano Comestible
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Mutagénesis Insercional
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Segregación Cromosómica
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Cruzamientos Genéticos
Idioma:
En
Revista:
BMC Plant Biol
Asunto de la revista:
BOTANICA
Año:
2012
Tipo del documento:
Article
País de afiliación:
Alemania
Pais de publicación:
Reino Unido