A large plasmid enables its host Agrobacterium tumefaciens to cause tumorous condition in a wide variety of dicotyledonous plants[see Ooms et al. Gene 14:33--50 (1981) )). The location and role of chromosomal genes in this phenomenon are not known. As the first stage in studying this aspect, a project was initiated to investigate the chromosomal genetics of the bacterium. R68.45, a P group plasmid, was chosen as a transmission agent. After a preliminary assessment it was decided to use C58 as a standard strain to carry out the mapping. The plasmid itself, as judged by the presence of antibiotic markers, appears to be stable in A. tumefaciens; its ability to promote chromosomal mobilisation, however, remains only in 60--80% transconjugants. Good Agrobacterium donors are capable of transferring chromosomal genes at a frequency varying between 10(-5) to 10(-6) per recipient. The recombinants are stable even under non-selective conditions. A linear linkage map consisting of 16 markers was built using coinheritance frequencies obtained from 21 four-point crosses.

A revised and enlarged genetic map of the Agrobacterium tumefaciens C58 chromosome has been produced with the help of plasmid R68.45. Apart from the location of several auxotrophic markers, the map also shows the position of two independent genes, ctul and ctu2, which, when mutated, block the tumorigenesis of the bacterium. Of these two, only ctul is complemented by the C58 chromosomal vir region cloned by Douglas et al. (1985). The same mutant was complemented by a chromosomal gene or genes located near leu of Rhizobium meliloti and known to affect the nodulation properties of that bacterium. It has also been observed that C58 tryptophan auxotrophs invariably lose tumorigenicity. Prototrophic revertants and mutants supplied with extra tryptophan for about two weeks after infection produce normal tumours. These investigations suggest that for successful tumorigenesis a continuous supply of tryptophan is needed (to be converted into auxin IAA?) at least during the early stages.

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