Time required for tumour initiation by Agrobacterium tumefaciens on pinto bean leaves.
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Biomedical subjects
Publications and source records attributed to J A Lippincott.
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The infectivity of Agrobacterium tumefaciens strain B6 irradiated with short-wavelength ultraviolet light was followed as a function of dose. Previously reported enhancements of B6 infectivity by ultraviolet irradiation, in samples inoculated after 1.75 hr of dark incubation at 27 C, or immediately following irradiation, were found to occur most frequently after losses in cell viability of 60% and of 90% or more, respectively. Changes in colony-forming ability and tumor-initiating ability with increasing dose showed no obvious correlation until the maximal infectivity promotion of samples inoculated immediately after irradiation was reached. Thereafter, both bacterial responses typically decreased in parallel. With low dose rates, infectivity promotions were obtained with less than 10% loss in cell viability. Data for tumor appearance and tumor growth resulting from inoculations with irradiated cultures showed no significant differences from controls, nor did the age of the bacterial culture or age of the host plant influence the response. The infectivity promotion appears to result from an increase in the proportion of viable cells that will subsequently initiate tumors. The characteristics of this ultraviolet infectivity promotion are shown to be most similar to those found in prophage and bacteriocin induction.
The ability of Agrobacterium tumefaciens to induce pinto leaf tumors may be enhanced two- to threefold after treatment with mitomycin C. The enhancement may be obtained with either lethal or nonlethal concentrations. With 10-min treatments, an optimal response was obtained with 0.005 mug of mitomycin C per ml in the absence of any change in the number of viable cells. Both the tumor induction process and the tumors induced by treated cultures appear qualitatively the same as controls. To account for these results, the antibiotic must increase the proportion of viable cells that will subsequently initiate tumors. One, or at most a few, random lesions in the bacterial chromosome seem to be the necessary requirement for this promotion. At mitomycin concentrations of 1 and 5 mug/ml, the ability of A. tumefaciens to initiate tumors is rapidly lost, indicating that a fairly intact bacterial chromosome is one of the essentials for the tumor induction process.
Lippincott, Barbara B. (Northwestern University, Evanston, Ill.), and James A. Lippincott. Characteristics of Agrobacterium tumefaciens auxotrophic mutant infectivity. J. Bacteriol. 92:937-945. 166.-Mutants of Agrobacterium tumefaciens auxotrophic for adenine, methionine, or asparagine are less infectious than the wild-type strain B6 from which they were derived and show increased infectivity on pinto bean leaves when the specific compounds required for growth of the mutants are added to the infected leaf. Reversion to a prototrophic form of nutrition is accompanied by increased infectivity. Tumors initiated by these auxotrophic mutants are shown to arise only at large wound sites where nutritional conditions may be less restricting. The data indicate that, after inoculation, the bacteria pass through a phase in which host-supplied nutrients are utilized for the production of one or more factors necessary for successful tumor initiation.
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