Authors:A. Zok, I. Forgács, A. Pedryc, R. Oláh, and E. Szegedi
. & Tinland, B. (2001): Transformation of tobacco plants with VirE1 gene derived from Agrobacteriumtumefaciens pTiA6 and its effect on crown gall tumor formation. Int. J. Hortic. Sci. , 7 , 53–56.
Authors:H. Aadel, R. Abdelwahd, S.M. Udupa, G. Diria, A. El Mouhtadi, K. Ahansal, F. Gaboun, A. Douira, and D. Iraqi
Although significant progress has been made on Agrobacterium-mediated wheat transformation, current methodologies using immature embryos as recipient tissues are labor intensive, time consuming and expensive. The use of mature embryos as explants is increasingly being recognized as an optimal method for developing regenerable cell lines during wheat transformation. Therefore, we have developed an Agrobacterium-based transformation protocol using mature embryos while adjusting several factors that influence genetic transformation efficiency. In this study, we focussed on acetosyringone concentrations, genotypes and different types of mature embryos (intact or longitudinally halved-embryos or fragmented into four latitudinal pieces) used as a source of explants for the genetic transformation. A. tumefaciens strain EHA101 harboring the plasmid vector pTF101.1 carrying the barley HVA1 gene and bar-selectable marker gene were used. Mature intact-embryos and longitudinally halved-embryos yielded the highest number of putative transgenic plantlets on the selection medium. However, no plantlets were obtained from latitudinal fragmented mature embryos. ‘Amal’ and ‘Rajae’ genotypes regenerated the highest number of putative transgenic plants and 200 μM acetosyringone was found to be the optimal concentration for their transformation. A total of 47 transgenic plants were selected with 11 plantlets showing resistance to leaf painting. Molecular analysis revealed that 1% and 0.66% of T0 regenerated plantlets were successfully transformed and carried the HVA1 gene for the ‘Amal’ and ‘Rajae’ genotypes, respectively. Additional analysis shows the transgene is stably inherited in the T1 generation. Based on the results, we conclude that among the influencing factors tested, genotypes, mature embryo explant types and acetosyringone concentration contribute significantly to the success of bread wheat transformation.
An efficient and reproducible protocol is described for shoot-bud regeneration and Agrobacterium tumefaciens-mediated genetic transformation of J. curcas. Treating the explants with high concentrations (5–120 mg/L) of TDZ for short durations (5–80 min) before inoculation culture increased significantly the regeneration frequency and improved the quality of the regenerated buds. The highest shoot-buds induction rate (87.35%) was achieved when petiole explants were treated with 20 mg/L TDZ solution for 20 min and inoculated on hormone-free MS medium for 30 days. Regenerated shoots of 0.5 cm or a little longer were isolated and grafted to seedling stocks of the same species, and then the grafted plantlets were planted on half-strength MS medium containing 0.1 mg/L IBA and 2 mg/L sodium nitroprusside (SNP). This grafting strategy was found to be very effective, to obtain that healthy grafted plantlets ready for acclimatization within 20 days. By the above mentioned protocol and with general Agrobacterium – mediated genetic transformation methods only 65 days were needed to obtain intact transgenic plants.
cultivar, leaf age and bacterial strain on transformation by Agrobacteriumtumefaciens. Plant Cell Tiss. Org. Cult. , 24, 115-121.
Effects of tomato cultivar, leaf age and bacterial strain on transformation by Agrobacterium
Authors:Nóra Gyémánt, Annamária Molnár, Gabriella Spengler, Yvette Mándi, Margit Szabó, and J. Molnár
Stachel, E. S., Messens, E., Van Montagu, M., Zambryski, P.: Identification of signal molecules produced by wounded plant cells that active the T-DNA transfer in Agrobacteriumtumefaciens . Nature 318 , 624-629 (1985