Laboratory Maintenance of Agrobacterium

Elise R. Morton1, Clay Fuqua1

1 Department of Biology, Indiana University, Bloomington, Indiana
Publication Name:  Current Protocols in Microbiology
Unit Number:  Unit 3D.1
DOI:  10.1002/9780471729259.mc03d01s24
Online Posting Date:  February, 2012
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Abstract

Agrobacterium species are plant‐associated relatives of the rhizobia. Several species cause plant diseases such as crown gall and hairy root, although there are also avirulent species. A. tumefaciens is the most intensively studied species and causes crown gall, a neoplastic disease that occurs on a variety of plants. Virulence is specified by large plasmids, and in the case of A. tumefaciens this is called the Ti (tumor‐inducing) plasmid. During pathogenesis, virulent agrobacteria copy a segment of the Ti plasmid and transfer it to the plant, where it subsequently integrates into the plant genome and expresses genes that result in the disease symptoms. A. tumefaciens has been used extensively as a plant genetic engineering tool, and is also a model microorganism that has been well studied for host‐microbe associations, horizontal gene transfer, cell‐cell communication, and biofilm formation. This unit describes standard protocols for laboratory cultivation of A. tumefaciens. Curr. Protoc. Microbiol. 24:3D.1.1‐3D.1.16. © 2012 by John Wiley & Sons, Inc.

Keywords: growth media; genetic analyses; taxonomy; opines; plant association; virulence; plasmids; attachment

     
 
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Table of Contents

  • Introduction
  • Growth Conditions
  • Common Antibiotics—Working Concentrations for Agrobacterium
  • Storage Conditions
  • Acknowledgement
  • LITERATURE CITED
  • Tables
     
 
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Materials

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Literature Cited

LITERATURE CITED
   Blanca‐Ordóñez, H., Oliva‐Garcá, J.J., Pérez‐Mendoza, D., Soto, M.J., Olivares, J., Sanjuán, J., and Nogales, J. 2010. pSymA‐dependent mobilization of the Sino Rhizobium meliloti pSymB megaplasmid. J. Bacteriol. 192:6309‐6312.
   Poindexter, J.S. 1978. Selection for nonbuoyant morphological mutants of Caulobacter crescentus. J. Bacteriol. 135:1141‐1145.
   Soberón, N., Venkova‐Canova, T., Ramírez‐Romero, M.A., Téllez‐Sosa, J., and Cevallos, M.A. 2004. Incompatibility and the partitioning site of the repABC basic replicon of the symbiotic plasmid from Rhizobium etli. Plasmid 51:203‐216.
   Tempé, J., Petit, A., Holsters, M., Montagu, M., and Schell, J. 1977. Thermosensitive step associated with transfer of the Ti plasmid during conjugation: possible relation to transformation in crown gall. Proc. Natl. Acad. Sci. U.S.A. 74:2848‐2849.
   Tzfira, T. and Citovsky, V. (eds.) 2008. Agrobacterium, From Biology to Biotechnology. Springer Scientific Publishing, New York.
   Winans, S.C., Kerstetter, R.A., and Nester, E.W. 1988. Transcriptional regulation of the virA‐gene and virG‐gene of Agrobacterium tumefaciens. J. Bacteriol. 170:4047‐4054.
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