E Keshamma, B. Madhusudhan, Department of Biochemistry, Center for Nano Science and Technology Research, Kuvenpu University, P.G. Centre, Shivagangotri Davangere 577 002, India; S. Rohini, M. Udaya Kumar*, Department of Crop Physiology, University of Agricultural Sciences, GKVK Bangalore 560 065, India; K. S. Rao, Centre for Ecological Sciences, Indian Institute of Science, Bangalore 560012, In...

In the present study high efficiency Agrobacterium tumefaciens mediated transformation system of wheat (Triticum aestivum L.) cv. Inqilab-91 was developed by exploiting bacterial culture density and acetosyringone concentration. Agrobacterium strain EHA101 harboring binary vector pIG121Hm, containing gene for GUS activity and hygromycin resistance was used in transformation experiments. Differe...

A procedure to enrich for auxotrophic and fermentation mutants of Agrobacterium tumefaciens is described. The method is based on the amplification of the killing power of carbenicillin by the addition of lysozyme. Isolation frequencies of some types of mutants are presented, with and without the application of the proposed procedure. The yield of mutants is usually enhanced a hundredfold per en...

Bacteria use conjugation systems, a subfamily of the type IV secretion systems, to transfer DNA to recipient cells. Despite 50 years of research, the architecture and mechanism of action of the channel mediating DNA transfer across the bacterial cell envelope remains obscure. By use of a sensitive, quantifiable assay termed transfer DNA immunoprecipitation (TrIP), we identify contacts between a...

A binary vector is a standard tool in the transformation of higher plants mediated by Agrobacterium tumefaciens. It is composed of the borders of T-DNA, multiple cloning sites, replication functions for Escherichia coli and A. tumefaciens, selectable marker genes, reporter genes, and other accessory elements that can improve the efficiency of and/or give further capability to the system. A supe...

Agrobacterium tumefaciens-mediated transformation is one of the most efficient and simple gene delivery systems for genetic improvement and biology studies in maize. This system has become more widely used by both public and private laboratories. However, transformation efficiencies vary greatly from laboratory to laboratory for the same genotype. Here, we illustrate our advanced Agrobacterium-...

Kinetic, microscopic, and biochemical studies show that virulent Ti (tumor inducing)-plasmid-containing strains of Agrobacterium attach to normal tobacco and carrot tissue culture cells. Kinetic studies showed that virulent strains of A. tumefaciens attach to the plant tissue culture cells in increasing numbers during the first 1 to 2 h of incubation of the bacteria with the plant cells. Five T...

Agrobacterium tumefaciens mediated transformation may offer a better alternative than the biolistic gun for genetic transformation of maize plants. This gene delivery system results in a greater proportion of stable, low-copy number transgenic events than does the biolistic gun, and is highly efficient. In the present work, we studied maize transformation using A. tumefaciens by identifying som...