Factors influencing Agrobacterium-mediated transient expression of uidA in wheat inflorescence tissue.
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.
Amoah
B K
BK
Biochemistry and Physiology Department, IACR-Rothamsted, Harpenden AL5 2JQ, UK.
Wu
H
H
Sparks
C
C
Jones
H D
HD
D016960:Agrobacterium tumefaciens / Q000235:genetics*; D004269:DNA, Bacterial; D005374:Filtration; D015870:Gene Expression; D005798:Genes, Bacterial; D005822:Genetic Vectors; D005966:Glucuronidase / Q000235:genetics; D018514:Plant Structures / Q000378:metabolism; D013010:Sonication; D014170:Transformation, Genetic; D014908:Triticum / Q000235:genetics* / Q000382:microbiology; D014618:Vacuum
D004269:DNA, Bacterial; C036483:T-DNA; D005966:Glucuronidase