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GrainGenes Reference Report: PRT-21-659

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Reference	PRT-21-659
Title	Factors influencing successful Agrobacterium-mediated genetic transformation of wheat
Journal	Plant Cell Reports
Year	2003
Volume	21
Pages	659-668
Author	Wu H Sparks C Amoah B Jones HD
Abstract	Summary: 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
Keyword	[ Hide all but 1 of 39 ] acetosyringone agrobacterium agrobacterium tumefaciens agrobacterium-mediated transformation bar bread cocultivation culture difference efficient fertile gene genetic modification genetic transformation genome hordeum vulgare inoculation mar media molecular analysis plant regeneration plant transformation plasmid progeny protocol ratios regeneration rice strain tissue transformation transformation frequency transgene transgenes transgenic triticum aestivum uida visual marker wheat variety

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