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Isolation of Agrobacterium Ti-plasmid regulatory mutants

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Summary

Crown gall tumors incited by Agrobacterium tumefaciens synthesize basic amino acid derivatives called opines. Opine production in tumours and opine catabolism by A. tumefaciens are coded by Ti-plasmids which confer oncogenicity on this bacterium. Catabolism of opines is inducible, and a method for isolation of regulatory mutants is described. From octopine-type bacteria, by plating on non-inducing substrates (noroctopine, noroctopine acid, D-histopine) we have isolated regulatory mutants of three types: constitutive, partially constitutive, and fully inducible by the analogue. From nopaline-type bacteria, by plating on octopine (a non inducing substrate) we have isolated analogous regulatory mutants.

Synthetic opines, in which the amino acid moiety has been replaced by toxic arginine analogues, are toxic for these regulatory mutants. We isolated mutants resistant to such synthetic opines, and found that some had lost the capacity to utilize octopine. A survey of a large number of such mutants revealed that all of them still incited octopine synthetizing tumors.

Mutants constitutive for octopine catabolism are in some instances also constitutive for Ti-plasmid transfer. A simple method for screening regulatory mutants for constitutive Ti-plasmid transfer is described.

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Authors and Affiliations

  1. Station de Génétique et d'Amélioration des Plantes, CNRA, F-78000, Versailles, France

    Annik Petit & Jacques Tempé

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  1. Annik Petit
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  2. Jacques Tempé
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Communicated by J. Schell

This work has been supported in part by grants from the Centre National de la Recherche Scientifique (contrats ATP 2814 and 3363).

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Petit, A., Tempé, J. Isolation of Agrobacterium Ti-plasmid regulatory mutants. Molec. Gen. Genet. 167, 147–155 (1978). https://doi.org/10.1007/BF00266908

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