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Appl Environ Microbiol. 1990 June; 56(6): 1782-1785
Copyright © 1990, American Society for Microbiology. All Rights Reserved.

Use of Ti Plasmid DNA Probes for Determining Tumorigenicity of Agrobacterium Strains

Thomas J. Burr*, John L. Norelli, Barbara H. Katz and Andrew L. Bishop{dagger}

Department of Plant Pathology, New York State Agricultural Experiment Station, Cornell University, Geneva, New York 14456

ABSTRACT

Probes consisting of T-DNA genes from the Ti plasmid of Agrobacterium tumefaciens were used for determining tumorigenicity of strains. Two 32P-labeled probes hybridized with 28 of 28 tumorigenic strains of the pathogen but not with 20 of 22 nontumorigenic strains. One probe, pTHE17, consists of all but the far left portion of the T-DNA of strain C58. Probe SmaI7 consists of SmaI fragment 7 of pTiC58, including onc genes 1, 4, and 6a and most of 2. Another probe, pAL4044, consisting of the vir region of strain Ach-5, hybridized with several nontumorigenic as well as tumorigenic strains. Colony hybridizations were done with 28 tumorigenic and 22 nontumorigenic Agrobacterium strains. About 106 CFU of the different tumorigenic strains were detectable with this method. Southern analyses confirmed the presence or absence of Ti plasmids in strains for which tumorigenicity was questioned. Colony hybridization with the T-DNA probes provides a rapid and sensitive means for determining the tumorigenic nature of Agrobacterium strains.

FOOTNOTES

* Corresponding author.

{dagger} Present address: Analysis and Identification Unit, California Department of Food and Agriculture, Sacramento, CA 95814.

Appl Environ Microbiol. 1990 June; 56(6): 1782-1785
Copyright © 1990, American Society for Microbiology. All Rights Reserved.





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