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GrainGenes Reference Report: JEB-52-1135

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Reference	JEB-52-1135
Title	Factors influencing Agrobacterium-mediated transient expression of uidA in wheat inflorescence tissue
Journal	Journal of Experimental Botany
Year	2001
Volume	52
Pages	1135-1142
Author	Amoah BK Wu H Sparks C Jones HD
Abstract	Summary: A critical step in the development of Agrobacterium tumifaciens-mediated transformation is the establishment of optimal conditions for T-DNA delivery into tissue from which whole plants can be regenerated. The efficient transformation of inflorescence tissue from 'Baldus', a commercial wheat variety, using the Agrobacterium strain AGLI harbouring the binary vector pAL156 is reported here. The effects of various factors on delivery and the transient expression of the uidA gene were studied including the duration of preculture, vacuum infiltration, the effect of sonication treatments, and Agrobacterium cell density. Optimal T-DNA delivery (as measured by uidA activity) was obtained from inflorescence tissues precultured for 21 d and sonicated. Increasing Agrobacterium cell density, the duration of inoculation/co-cultivation, and vacuum pressure, up to a threshold, increased uidA expression. The investigation of factors that influence T-DNA delivery is an important first step in the utilization of Agrobacterium in the transformation of immature wheat inflorescence tissue
External Databases	Pubmed: 11432931
Keyword	[ Hide all but 1 of 44 ] agrobacteria agrobacterium agrobacterium tumefaciens arabidopsis thaliana beta glucuronidase binary vector cell cell density cell-density commercial variety delivery density development efficient transformation establishment gene gene expression gene transfer gus high-efficiency immature inflorescences infiltration inflorescence microprojectile bombardment oryza sativa plant regeneration plasmid vectors regenerative ability reporter gene somatic embryogenesis sonication strain t dna t-dna tissue transformation transient transient expression triticum aestivum uida uida gene vector wheat inflorescence wheat varieties

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