PMC

Expression of virB and virE in C58, Δ1392, and Δ1392 with a virG(Con) locus (N54D). A. tumefaciens strains containing a virB::lacZ fusion or virE::lacZ fusion were incubated in induction broth with arabinose substituting for glucose for 24 h ± acetosyringone, and β-galactosidase activities were measured as described in Materials and Methods. (A) virB expression; (B) virE expression.

Virulence assay on Kalanchöe leaves. Wild-type (C58), Δ1392, Δ4851, Δ5306, Δ5307, and A136 (a negative control) strains were grown in MG/L media as described in Materials and Methods, and 1 μl of each culture was used to inoculate Kalanchöe leaves. Photos were taken 30 days after inoculation.

Functional complementation of A. tumefaciens CS in Sinorhizobium. The vectors, each containing one of the cloned putative CS genes, were introduced by electroporation into S. meliloti Δ1A, a mutant with a mutation in CS. The cells were plated on MM NH₄ medium with arabinose and kanamycin. Colonies were then streaked out onto MM NH₄± arabinose. Growth in the absence of arabinose indicates that the cell has a functional CS.

Growth curve of Δ1392 and wild-type (C58) in AB minimal medium with various supplements. Log-phase cultures of C58 and Δ1392 were inoculated into AB minimal medium with the various supplements indicated with a starting OD₆₀₀ of 0.001. Cells were grown at 28°C. Optical density at 600 nm was measured at 2-h intervals over a 36-h period. C58+Ara, C58 was grown in AB minimal medium with 0.4% arabinose as the sole carbon source; C58+Gluc, C58 was grown in AB minimal medium with 0.4% glucose as the sole carbon source; Δ1392+Suc+Gluc, Δ1392 was grown in AB minimal medium with 0.4% succinate and 0.4% glucose as carbon sources; Δ1392+Gluc+Glut, Δ1392 was grown in AB minimal medium with 0.4% glucose and 0.4% glutamate as carbon sources.