Factors influencing Agrobacterium‐mediated transient expression of uidA in wheat inflorescence tissue

Abstract

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.

Purine synthesis and increasedAgrobacterium tumefacienstransformation of yeast and plants

“…The most striking results were observed with inhibitors that block both purine and pyrimidine synthesis (azaserine and acivicin). The magnitude of this effect is many times greater than similar effects reported in previous studies (25,(30)(31)(32)(33). A less-pronounced effect was observed for the purine synthesis inhibitor mizoribine, whereas the pyrimidine synthesis inhibitor 5-fluorouracil had no effect.…”

Efficient production of transgenic plantls of Vanda through sonication-assisted Agrobacterium-mediated transformation of protocorm-like bodies

“…In Agrobacterium-mediated transformation, wounding treatments such as sonication Finer 1997, 1998;Amoah et al 2001;Tang 2003;Zaragoza et al 2004;Gaba et al 2006) and scratching (Hoshi et al 2005) have been used to enhance transformation in various species. In the present study, all of the 3 kinds of wounding treatments increased the transformation efficiency by 2-5 folds in comparison with the control and the highest transformation efficiency was obtained by sonication treatment, which gave ca.…”

Establishment of Agrobacterium-mediated genetic transformation system in Dahlia

“…Higher concentration of bacteria, i.e., less dilution of Agrobacterium liquid culture (OD 600 =1.2) with 40 ml of liquid MS medium (1 : 20 ratio), generally gave more GUS positive MSP (1.3-2.6 times as high) than that diluted at 1 : 100 ratio, except in the case where sonication was not applied and co-cultivated at 25°C. In Agrobacterium-mediated transformation, sonication has been used to enhance transformation in various species (Amoah et al 2001;Shrestha et al 2007;Tang 2003;Finer 1997, 1998;Weber et al 2003;Zaragoza et al 2004). Similarly, in the case of dahlia, sonication increased transformation efficiency in comparison with the untreated control (1.4-2.6 times as high), except for the case where dilution of bacterial suspension was at 1 : 100 ratio and co-cultivated at 25°C.…”