Conjugative transfer of pTd33-plasmid between strains of Agrobacterium

Supramembrane structures that connect conjugating agrobacterial cells were visualized for the first time by transmission electron microscopy. The primary contact of cells during conjugation was shown to occur through the formation of long pili containing no VirB1 protein. Pretreatment of agrobacterial cells with acetosyringone resulted in a six- to tenfold increase in the transfer frequency of the plasmid pTd33 at 19-25 degrees C and had almost no effect at 30 degrees C. The transfer of the plasmid pTd33 from A. tumefaciens strain GV3101 to plasmid-free A. tumefaciens strain UBAPF-2 was 16 times decreased after the centrifugation of cells. The transfer efficiency of the plasmid pTd33 from A. tumefaciens strain LBA2525 (virB2::lacZ) to plasmid-free A. tumefaciens strain UBAPF-2 was one order of magnitude lower than the transfer from the wild-type A. tumefaciens strain GV3101. Treatment of donor cells with 0.01% SDS before mating decreased the transfer efficiency by a factor of 26. The role of pili in the establishment of contact between conjugating cells of agrobacteria is discussed.