Factors influencing successful Agrobacterium-mediated genetic transformation of wheat

H. Wu, C. Sparks, B. Amoah, H. D. Jones

Allbwn ymchwil: Cyfraniad at gyfnodolynErthygladolygiad gan gymheiriaid

Crynodeb

The development of a robust Agrobacterium-mediated transformation protocol for a recalcitrant species like bread wheat requires the identification and optimisation of the factors affecting T-DNA delivery and plant regeneration. We have used immature embryos from range of wheat varieties and the Agrobacterium strain AGL1 harbouring the pGreen-based plasmid pAL156, which contains a T-DNA incorporating the bar gene and a modified uidA (beta-glucuronidase) gene, to investigate and optimise major T-DNA delivery and tissue culture variables. Factors that produced significant differences in T-DNA delivery and regeneration included embryo size, duration of pre-culture, inoculation and co-cultivation, and the presence of acetosyringone and Silwet-L77 in the media. We fully describe a protocol that allowed efficient T-DNA delivery and gave rise to 44 morphologically normal, and fully fertile, stable transgenic plants in two wheat varieties. The transformation frequency ranged from 0.3% to 3.3%. Marker-gene expression and molecular analysis demonstrated that transgenes were integrated into the wheat genome and subsequently transmitted into progeny at Mendelian ratios.

Iaith wreiddiolSaesneg
Tudalennau (o-i)659-668
Nifer y tudalennau10
CyfnodolynPlant Cell Reports
Cyfrol21
Rhif cyhoeddi7
Dyddiad ar-lein cynnar16 Ion 2003
Dynodwyr Gwrthrych Digidol (DOIs)
StatwsCyhoeddwyd - 01 Maw 2003

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