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Journal of Bacteriology, July 2000, p. 3705-3716, Vol. 182, No. 13
0021-9193/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.

Genetic and Environmental Factors Affecting T-Pilin Export and T-Pilus Biogenesis in Relation to Flagellation of Agrobacterium tumefaciens

Erh-Min Lai,1,dagger Olga Chesnokova,1,Dagger Lois M. Banta,2 and Clarence I. Kado1,*

Davis Crown Gall Group, University of California, Davis, California 95616,1 and Department of Biology, Haverford College, Haverford, Pennsylvania 190412

Received 23 September 1999/Accepted 18 April 2000

The T pilus, primarily composed of cyclic T-pilin subunits, is essential for the transmission of the Ti-plasmid T-DNA from Agrobacterium tumefaciens to plant cells. Although the virB2 gene of the 11-gene virB operon was previously demonstrated to encode the full-length propilin, and other genes of this operon have been implicated as members of a conserved transmembrane transport apparatus, the role of each virB gene in T-pilin synthesis and transport and T-pilus biogenesis remained undefined. In the present study, each virB gene was examined and was found to be unessential for T-pilin biosynthesis, except virB2, but was determined to be essential for the export of the T-pilin subunits and for T-pilus formation. We also find that the genes of the virD operon are neither involved in T-pilin export nor T-pilus formation. Critical analysis of three different virD4 mutants also showed that they are not involved in T-pilus biogenesis irrespective of the A. tumefaciens strains used. With respect to the environmental effects on T-pilus biogenesis, we find that T pili are produced both on agar and in liquid culture and are produced at one end of the A. tumefaciens rod-shaped cell in a polar manner. We also report a novel phenomenon whereby flagellum production is shut down under conditions which turn on T-pilus formation. These conditions are the usual induction with acetosyringone at pH 5.5 of Ti-plasmid vir genes. A search of the vir genes involved in controlling this biphasic reaction in induced A. tumefaciens cells revealed that virA on the Ti plasmid is involved and that neither virB nor virD genes are needed for this reaction. The biphasic reaction therefore appears to be mediated through a two-component signal transducing system likely involving an unidentified vir gene in A. tumefaciens.

* Corresponding author. Mailing address: Davis Crown Gall Group, One Shields Ave., University of California, Davis, CA 95616. Phone: (530) 752-0325. Fax: (530) 752-5674. E-mail: cikado{at}ucdavis.edu.

dagger Present address: Department of Biology, University of Michigan, Ann Arbor, MI 48109.

Dagger Present address: Department of Plant and Microbial Biology, University of California, Berkeley, CA 94720.

Journal of Bacteriology, July 2000, p. 3705-3716, Vol. 182, No. 13
0021-9193/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.


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